’ 7 Al Ra eat he op Doth » 7 Rig Fete oh tat ane <- we Ae ae Py + Ea Oy ee pl vappet © wT B ‘CHE: BULLETIN OF PEE COLLEGE OF AGRICULTURE, Toxyo IwrerRiAL UNIVERSITY, JAPAN. Vou. Vie 1902—1903. s _ , ‘e- re 4 Cm \; “4 <\e 17 4 cM £ £ Ae ee - . a 2 42 ee 10-with rund from Univerbity of Toronto | * C ji e # mat | Shana http://www.archive.org/details/bulletin5\ ee. os ea CONTENTS @F°OVOLRUME V. KITAO: Inwiefern kann man das Holz als einen isotropen K6rper betrachten? - = = s = = ~ = 2 . 2 5 . IKEDA: Studies in the Physiological Functions of Antipodals and Related Phenomena of Fertilization in Liliaceae. I. Trycirtis Hirta. - - - K. TOYAMA: Contributions to the Study of Silk-Worms. I. On the Embryo- logy of the Silk-Worms. - - - - = = z = N. NIVTA: Ueber das wirksame Princip des Tuberculinum Kochii. - - M 5. O LOEW und Y. KOZAI: Ueber Ernahrungsverhaltnisse beim Bacillus Prodigiosus. - - - - = - = == - - . TOYONAGA: Ueber die Vertheilung des Kalks in Thierischen Organ- ismus. - - - - - - - - - - - - SAWAMURA: On the Digestive Porer of the Intestinal Canal. - - . LOEW and S. SAWA: On the Action of Manganese Compounds on Plants. = = = 2 = = a = aM = e J OSCAR LOEW: Ueber die Wirkung des Urans auf Pflanzen. - . - K. ASO: On the Physiological Influence of Manganese Compounds on Plants. . ASO: On the Action of Sodium Fluorid upon Plant Life. - - - . ASO: On the Action of Sodium Silicofluorid upon Plants. - - - SUZUKI: On the Action of Highly Diluted Potassium Iodid on Agri- cultural Piants. SUZUKI: On the Poisonous Action of Potassium Ferrocyanid on Plants. - . ASO: On Oxidizing Enzyms in the Vegetable Body. - - - - S. SAWAMURA: On the Curing of the Kaki Fruit. - . - - - .. ASO: On the Different Forms of Line in Plants. - : - s = .. TAKAHASHI: On the Alcohol Production in Phzenogams. = = = . SAWA: Can Alcohols of the Methane Series be Utilized as Nutrients by the Green Plants ? = = es = : é u Z Z . KIMOTO: On the Occurrence of Mannan. = = = e < s . KATAYAMA: On the General Gecurrence of Bacillus Methylicus in the Soil. - - - : = : : : : = zy f fs . SAWAMURA: On the Liquefaction of Mannan by Microbes. — - - - .. SUDA: Chemical Note on a Singular Pheenogamic Parasite. - - - . SAWAMURA: On the Action of Formaldehyd on Pepsin. - - - . SHISHIDO: Ueber die Einwirkung des Hara-Brennens. - - - . HEFELE: Die zukiinftige Bewirtschaftungsform des Japanischen Waldes ! . HEFELE: Wald und Wasserwirtschaft. - - - - - - el Lol ~\ _ = Oo Ow ~ ~~] “I NN ™N G9 _ bo wo YI we ta —< > 1 7) + vo ve ui Ww -« [ ii ] H. SHIRASAWA: Ueber Entstehung und Verthzilung des Kamphers im Kampherbaume. - - - - - - - - - - - S. SAWAMURA: Investigations on Flacherie. - - - - - - O. LOEW und Y. KOZAI: Zur Physiologie des Bacillus pyocyaneus, I. - M. TOYONAGA: Ueber den Kalkgehalt der Milchdrise. — - - - - O. LOEW: Der Erntequotient. —- - - - - - - - - O. LOEW: Ueber die physiologische Wirkung des Chlorrubidiums auf Phanerogamen. - - - - - : - - - - - M. NAGAOKA: On the Stimulating Action of Manganese upon Rice. - - S. SUZUKI and K. ASO: On the Physiological Action of Iodine and Fluorine Compounds on Agricultural Plants. K. ASO: On the Chemical Nature of the Oxidases. - - - - - S. SUZUKI: Can Sulfo-Derivatives of Hydroxylamine Serve as a Source of Nitrogen for Plants? - - = - - - - - - - K. ASO: On the Influence of a Certain Ratio between Lime and Magnesia on the Growth of the Mulberry-Tree. - - - - - - G. DAIKUHARA: On the Influence of Different Ratios between Lime and Magnesia upon the Development of Phaseolus. - - - - - G. DAIKUHARA: On the Behavior of the Phosphoric Acid in the Soils towards Different Organic Acids. - - - - - - - - M. NAKAMURA: Can Boric Acid in High Diution Exert a Sfimulant Action on Plants ? - - - - - - - - - - - S. SUZUKI: On the Action of Vanadin Compounds on Plants. - ; - S, SUZUKI: Can Potassium Ferrocyanid Exert any Stimulant Action in the Soil on Plant Growth? - . - - - = Z 4 y 2 S. SUZUKI: Are Soluble Iodids Absorbed by the Soil? - - . - - In wie ferne kann man das Holz als ein isotroper Korper betrachten ? VON Prof. Dr. D. Kitao. Wir wollen jetzt die Componenten der elastischen Druckkrafte+ mittlst der Neumann’schen Methode* Ny YZ Xx, Y,Zy also Functionen der Verschiebungscomponenten u, v, w darzustellen suchen und dabei untersuchen, wie ein. krystallinischer Korper, wie Holz als ein isotroper KGrper betrachtet werden kann. Die Krafte welche die Theilchen eines dilatirenden K6rpers in ihre Gleichgewichtlage zuriickzufiihren streben und als Driickkrafte wirken haben ihren Grund in den Kraften, welche zwischen den Theilchen wirken und so beschaffen sind, dass sie nur dann wirken, wenn die Entfernung der Theilchen klein ist und die Theilchen selbst aus der natirlichen Gleichgewichtlage verschoben worden sind. Unter dem nattirlichen Zustand eines Ko6rpers verstehen wir den Zustand, dass ohne Einwirkung dausserer Krafte alle im Jnneren des Korpers thatigen Krafte sich gegenseitig aufheben. Es seien die Coordinaten eines Massentheilchen dm z y z, die des zweiten Massentheilchen dm aber ++ y+y7 2+€, und es wirke auf den Punkt (xyz) die Kraft am auf (0) langs der Linie e= VE +A OE die Resultate aller auf dm wirkenden Krifte ist =dm \/ (0) ane! wo das Integral so weit zu erstrecken ist, als 7(o) nicht verschwindet. Es ist sonach im naturlichen Zustande * Franz Neumann: Theorie cer Elasticitat pag. ’82. 2 D. Kitao: o=am | am f ( p= == =an\ au f ( a an\ am’ f ( py Nun erleiden die Theilchen (yz) und x+&, y+y, z+€ relative Verriic- kung, deren Componenten durch bezeichnet werden mégen. _Die Componenten der auf dm ausgetibten Molecularkrafte sind Eng p+’ am | A di! =din\ amit (9 +, o')\——= ait | B dn =dm\ aim'f (po fa pee Par 2 am \ Ec dn! =dm\ nif (p+ ae wo p’ durch Pt+PhH=V(ETSV + Qty yP+Otey definirt ist, oder mit Vernachlassigung unendlich kleiner Grésse héherer Ordnung (meyer =| ay a ! pep=so+ngt+oe mit demselben Grade der Anniherung geschrieben werden kann F(p+e') (25 )=7( ner (rep) Pp =(s(—) +07) G42) (1-4) =$ (0) O34 4 ( (9 - LP oe Bas =~ (9) + Ae 4. Hip) 2 Wo (2) 1g) = ee ee Hp) 0 do gesetzt worden ist, so folgt. am | A adm'=dmn | an (= 1 (p)+ L a 2 E' +e Fp) v0’) dm | B din'=dm | an (2r(—)+ (0) + Hy 7' +7 F(p) e 0") In wie fern Kann man das Holz ete. 1S) In wie fern kann man das Holz. am \ T dm'=dm | an (£7) eee F(p) oe!) i i Es werde ferner auf die Oberflache des Kérpers ein Druck ausgeiibt, dessen Componenten fiir die Flachenheit BEHZ heissen. Es seien ferner 0% dv dw unendlich kleine virtuellen Verriic- kungen du Ov Ow ihre Werthe an der Oberfliche, deren Elemente dw Sein mége, das Princip der virtuellen Verriickung giebt. \ (E dv+H 0v+Z dw) do +\ | adm' A ou + [an] am!' B ow at an | aim’ T dw=o | aim \ aiu' A Ou nehmen wir eine wichtige Umformung vor. In diesem Integral kommt Mit dem Integral die Wirkung des dm auf dm’ als diejenige des dm! auf dm vor, und beide Wirkungen unterscheiden sich im Vorzeichen. Die Coordinaten até y+ s+ verwandeln nach der Verschiebung z, v, w in Out » Ou Ou OREO Tern Gare Gagan = Ov Ov Ov Bite / ak A mae a AWA Ow. Ow ow Oe Da ¢ 7 € unendlich klein sein sollen, die Gréssen S+E+am+ 4 D. Kitao: sind nichts anderes, als die Componenten der Verschiebun g, welche die beiden Punkte + y z, und ++¢ y+7, s+ nach der stattgehabten Ver- schiebung gegen einander erhalten haben d. h. abt tog cS / aie tng +e C= go ne ee Es ist so dann | mz) p ‘=| amp) (<¢ — r+ : —— Pog (= 0s (Say) # (Get oder indem wir setzen ? haa=fadn'y Zip) has dn'gs ist Iya \dn'¢s asia hs = lange t = oe ul Aare \am G ri Wo ; An=dy As=tn Ag=dy sind, so folgt =) In wie fern Kann man das Holz ete. Ut Dehnt man das Integral tiber den ganzen Ké6rper aus, so ist es ofienbar dass wenn wir den Coordinatenzuwachs fur dm mit abc, denjenigen fiir dm’ mit a’ 8’ c' bezeichnen ahaa A du= sf fan am A (da—da')= sam adim' A d(a— a’) =3{ {am dm! A (0%') da ja J=a—a y=b—b' Cl=c—e sind, wir erhalten somit jee (= du+H dv+Z dw) » ~ 4+- | am \ du! A 03! ds f in| ain' Bay! AI am \ am' Teg! oder in dem wir fir A BT ihre Ausdrticke einsetzen [ (= 0u+H 6v+Z dw) dw v 0 FN ao \ +H\ am ane LAP) (gga +707! + €0£') i +a | am ait! Zin) (C'0S' +709! + C0") ~ +alfen dm' V (~) po! (S05! +404! + fae')=0 Da nun aber 0 00')=S05' +707' + £02'=pdo' so folgt | 20 (21 du + Hdo+ Zoe) ae fj am am! FAO op) aA Be dint L- (9) (6? +77 +€") ? 6 D. Kitao: ae mall dm dam F(p) pe" =O Es sind die Verschiebungscomponenten des Punktes + y s, u v w sodass seine Coordinaten nach der Verschiebung Btw PUM SP w. sein werden. Es gelangt der Punkt ir rtuté' ytot7 stu+l’ Es ist so dann jens p =e thege tinge Ov Ov Ov + hia thie + hos ae ye fe Me a) oe oy E Da ferner aint ZO) P4740") = j dnt EP) ge (BE) +(32) ye (+ Ou, Ov Ov Ov 4 ow Ow Ox Oy Of Oy on aa a) ay) aay “(( al Ou Ou reed Ov ov 4 Ow Ow ( ( +257 1 27\ Oy Os * Oy Oz * Oy’ Of =) +(22)+ (ey) Ou . Ov ow Ow Se Bet Oe Ge ae ee Me & @ a da ferner In wie fern kann man das Holz ete. 7 +2hal Ou Ou Ov Ov. Ow =) oy SOds | Op Oa Oy Bz Ou Ou pee 2O0u Ou 2 =n con't =8(2( 52) +0 (Se) +0( Be) +, ey Z Ox ae Ou Z moe ie cy ee? i) af Ov-\" Ov ov \2 Ov Ov . wef (22) ay (8 By ) oe () ps Seer lov Ov, Ov Ov ee, Oz’ yh , Ou Ou. Ou Ow ge Ou Ss) ,Ov Ow ae a = ae Ou w ‘oy’ az Cae Os g Ou Ow, pov Ow | 2 dv =) G2 as." Oy ¥oe ">. Os Os indem man die Multiplication ausfiihrt, und nach dem & und 7 und etc ordnet. Ou Ov Ow =§(ar) +0(Se y+ ee) D. Kitao: i ihigot Vin. | GON Ws @d) 208) = Oe ~ Ov tg i(s) +(e 425, oy 25e oe) Bee ee owl On Ow Ou Ow £2 2 Pe sae Noch : Sta Bet ace: Pees +ee(S +(2) PREP Os We De = ) onn/ ( OY Vig f OW ow Ov, Ov Ow +t0((Sz) +( ay) +235 By toy Be) + &; i Ou 5 Oe Ov NOx oy one = Cu Ou Ou Ow £3 evict \ = eds Perera =C res 460 25%, en ert =) son OF Ov Ow 3 - +825. Oy Wis: a Ow Ow Ou Ow 35 SAE wee +O8(ae Oz? aa’ = Ow Ow Ov Ow ie Pe bsdieaiiriacieatys Be fse bes SS 4Oy 250, an? Cee 5) con |, C8) GL. ou ag Ou Ow Ov Ow +P 25 ae +2, OY | “Oy on) oauer Ou Ov Ou Ow +? Oe? Os 235 By) pe 4 Oe Ov Ou Ou Ov Ou Ow +Ht(2-5e Be +25 ‘Oz 1 Oy’ Oy Oy’ Oy pi CRS tng, SY ) “OP OF Ox’ Oy oe |. OULOW . CbaaD Ow Ou Ou Ow Tas 2Gs Gz | Gy es Cee Ou Ow Ov Ow mPae? Oe tas) ae) Setze-man ferner 2a, = [an i (p) és 204 = fan F (p) #7 2a; = [am F (p) &y 2 24= farm F(p) 7! 2ay= (dm F(p)t! adn! F (0) 7°€ say | di F (p) CS a F (p) "4 2at= {dm F (p) &% zai {dn F (p) C7? 2an= ain’ F (p) €°6? 2a [dn F (p) &7? In wie fern kann man das Holz ete. . F (0) 54 ed F(p) 69°C 2a {di F(p) 590° fam ree an( Se) 220342028) sane (2028) (O2Y 228) 0B) tan 32428) 4238 22) san ((ME BY 12% Bu) aS ee) Ou Ov\f(ou ow oufow ov +a (Se+52)(Se+S2 )+se(Se+35)) O 2 n a) a Ov (= +34)) Ou . Ov +4a,( (2 +5~\( y OZ Oy\ ex ' Oz Ov . ow Ou Ow (Ou . Ov + dau (3e+ +o) es o+52)\+or(o-+55)) Man setze ferner 99=dan\an' pO (gn +7?+€'") so erhalt man iP jam Oe 44 ree odu Odv O0s + haa + ha ss te ae " 7 2ow aes ip 1,032 ae ae Re pe) Setzt man ferner 2 =Ay tks + Ay - ‘ta — : GP rn - Mane, “22 Oy 23 Oz vs= hyo + 21 — ow ow Ow y= hyo aaa oe, EGS: 7 w= haya + here + vee Ww =A, S= +2 Se + hase So erhalt man * Oou Oou Oou 60 — [atm (1 + + ul — DG Om ee — mes Oow Oow Oow ss Eo a ey + WW e) Setzt man endlich Ou 6) wU @) va On=an> + aya Sy a a ta( 5242 SEER Oy 2) 5 a(S + a) e) ) va aa Qn =an5— +a: Se awe +4; = a= S) +a au 2 =e ge) x Ou +a + =) Op = ayo Faw ge + tego +a,(3 Ow =) 4-45 (= +54) oy Or | Os Ou , O tau Sy 32) Ou ie) ) Z rau) y= N= ays 5— + aya 5 + az = +a ov ges 2 2) + an Se : + os Oo Ox +an( $e “Oy = Or Ou Os Ov ) ) Se In wie fern Kaun man das Holz ete. it Ou Ov Ov . Ow Ow . Ou 0O13=0O3= arse tg, +465 ae (e+ Ze) tal S +54) a4 Ou . ov oa 5) Ou Ov Ou Ov . ow Ow u Qp= Oy =a + LE + Fis tal S-+5 Jra( Se +5") + ay Si +32) y x So erhalt man x OOu Cou Oou 0 R= dn (Ou Se + ye +0, Oov Oov Oov + Oo, Or + One ) ar Ons O02 O07 Oow +0: Ge + Dae + Osan) Wir erhalten somit als Gleichgewichtsbedingung | — 0u+H 0v+Z dw) +40P+10+30R=O Wir untersuchen die letzten drei Integrale besonderes, indem wir setzen din = pdxrdsdz = pdt und schreiben in dem Integrale Nome Cou Opa,du Opdy g (hu 0P= a dop( dn cos ma +A. Cos mY +A3 COS NE ) i +(ii COS WA + Aj yg COS 2) + Asg COS NS oe 12 D. Kitao: cos 2x +Ay Cos ny +133 cos nz ee +(% -f«|2 hn i Oe +1 Ox Oy O# eenae OMAsy Oph g Ox Ver r Os Opa 4 Ofthss mes Ofthss | Ox toy oz | Wobei das Oberflachenintegral tiber die ganze Oberflache des K6rpers Vv tae auszudehnen ist. Die Grossen 4 sind im Jnneren des Korpers constant. Wir kénnen sie aber .nicht unmittelbar an der Oberflache constant vorstellen, da in dem Integrale 2 um so mehr Glieder fehlen, je naher der Funkt xyz an der Oberflache liegt. Die Grésse 4 muss also so beschaffen sein, dass sie, da der Wirkungskreis der Molecularkrafte sehr klein ist, zuerst sehr rasch variirt mit wachsender Entfernung von der Oberflache rasch gegen eine constante Grenze convergirt. Das Raumintegral braucht daher nur itiber die der Oberfliche unendlich nahe liegende Theile ausgedehnt zu werden, wenn der K6rper homogen ist, Das Raumintegral denken wir uns in zwei Teile geteilt, eins tiber den Theil des Kérpers ausgedehnt, innerhalb dessen A Constant ist, das andere iiber den tbrig bleibenden Teil in dem letzten Integral setzen wir dt=dw x dn dt=dwx dun wo dz ein Element der Normale x bedeutet, yA ist demnach eine Func- tion von z, w. Bei der Kleinheit der Grenzen von x k6nnen wir wohl ohne Bedenken, mit Neumann annehmen* dass pA eine Function von z allein sei, eine Annahme, die nichts anderes aussagt, also dass pa in derselben Tiefe unter der Oberfliche iiberall denselben Werth habe Wir k6nnen, in sofern dieses erlaubt ist, setzen e) pa _ Opa sacs ——COs (77.7) so dass wir schreiben k6nnen uh n ) | open } -| dw | an oe Ou COS nx. 0 * Theorie der Elasticitit 96. In wie fern kann man das Holz ete. 13 wo 2z=o0 der ausseren Oberflache entspricht, Particulare Integration ergiebt = — ao OupAy, COS (na) aw (tpn +{ dw ouph,, cos (2x) Mithin erhalten wir of = — fe wy (a cos 74 +A, Cos my +A, Cos (25 ‘i + (da Cos 24 +d» COS 1 + hog cos (uz +(2n cos 27x +g COS 2y+ Ag COS (125 a _ , Lou cos (722) +{ aw yan (4: Zou COS NX bg ja cos 7y jg SH C08 (xe \ ai an an ) , @OV COS nN dovcoszy , , sao cos =e 44, —————_ (Oo aaa a FAs an + An adn Bi ee) 7 aow Cos nx 7 dow Cos zy my = cos (zs Se 2 an as an an — [ras (An ae + dry of a = )-+ete Die Dichtigkeit # kann innerhalb der Molecularentfernung sich anderen, Wir wollen den Kérper so beschaffen vorstellen, dass eine Volumen- einheit tberall dieselbe Menge Korper enthalte, d. h. so ist die mittlere Dichtigkeit iiberall constant und der K6rper ist homogen gebaut da ferner A in dem ersten und dritten Integral wesentlich constant=o, so verschwinden die Integrale. Wenn wir ferner die Annahme machen, dass man von dem Zustaénden der Theilchen die unendlich diinne Schichte der K6rperoberfliche absehen kénne, dass die Theilchen dort tiberall dieselbe Verriickung erleiden, als wenn die besagte Schichte starr ware. So ist die Grosse wie du cos (w+) von x unabhingig, so verschwindet das zweite Integral. Mithin ist itberall 0P=0 in so fern als der Koérper homogen ist, und man von den Zustanden der Theilchen in der Oberfliche abstrahiren kann. Selbst aber, wenn der Koérper heterogen ist, so verschwindet 0P, da w Gréssen Ay etc in dem inneren Punkt verschwinden, wenn es nur von den Vorgingen in der Oberflachen- 14 D. Kitao: Fir den Ruhezustand des Ko6rpers in schichts abgesehen werden kann, h. ohne Aiissere Krafte muss ¢P seinem natiirlichen Gleichgewichte d. unter allen Umstiinden verschwinden, dann wenn =©0 =] 0 20 so miissen auch L=C.= 7 >=>w—e da nun du dv, dw ganz willkiirliche Functionen sind, so wohlim Inneren als an der Oberfliche des KGrpers, so mussen cays ie} e) An 5 + Ay a = Aygo =0 ete Ay CoS (22) + Ay CoS (zy) +A Cos (uz) = 0 etc d.'h. aus den letzten drei Gleichungen folgt, allgemein Dap = Aya =Ay3 = Ag = dog = hog = Ag: = 4 = 43 = 9 wodurch auch die drei ersteren Gleichungen befriedigt werden. Um 0Q zu untersuchen nehmen wir dieselbe Transformation vor, in dem wir setzen i Odu Opi,du Oum, o = CC Oo Ox Ox Or Mithin 00= — | Law| uw, cos ux +u, Cos ny +u3 Cos (25) Jor +(%, V, COS n24 4- Vs cos ny +r, V3 cos (vs (o2) ie +( w, cos 2x +. w, cos ny + Ww; 3 COS (25 2) Bao Ouu, , Opt, , Oftits \n —( ae (2H 3.) \ \ OF q oy Os pu ON, , O"tU. , O"va He ett ae ye Opw, , OPW, OKs Nay +(- Oa. hie weed das Raumintegral braucht nur itiber die der Oberflache unendlich nahe da m w#, uw, etc. fiir jeden innern Punkt Schichte ausgedehnt zu werden, verschwinden. In wie fern Kaun man das Holz ete. 15 Wir denken uns an der Oberflache eine unendliche diinne Schichte von der Dicke z innerhalb deren 2 von o sich unterscheidet, und aus dieser Schichte ein Prisma ausgeschnitten, dessen Grundflache dw unend- lich klein, aber unendlich gross gegen die Hdéhe ist, dieses Prisma soll in seinem Theilchen tiberall dieselbe Verriickung erleiden d. h. wie ein starrer K6rper sich verhalten. Wir bilden dieses Integral iiber dieses Prisma und vernachlassigen dabei die vier Seitenflachen gegen die beiden Grundflachen da nun 4# nur von x abhangen soll und in jedem Theil des Prismas “ v w dieselben Werthe haben, so hiaingt yw etc nur von x ab. Es ist daher Of, _ ie, Opts, On Und wenn wir wieder setzen erhalten wir Opt, , Oftuts | Oftils ) . =e ay ee he - ek va pele aie Aplus oN = ao ian os cos 7a + _ 2 cos nyt oF cos wz Ou =| dw rm uy COS NX + My COS 2p + 2% COS nz) das erste Glied bezieht sich auf die Oberfliche das zweite aber auf die innere Grenze, weil weder cos etc noch du dv dw von x abhingen soll. Bildet man weiter auf dieselbe Weise die Ausdriicke eee OND, , 2128 a “" Ox Oye Ory ~ (GHW , Opis , + Sess \« ( Or Ee ay . Os Ow so erhalt man 16 D. Kitao: ~ Pat ae = Ta MN 10={p aol my Cos #4 +22 COS NY + Uz COS NS ew oY +a aol % V, COS NX +Vy COS NV+ COS ne ae v > ES. x +a dol % COS 2.4 + Wz COS ny +3 COS NS - Wie cu Wo die Groéssen sich auf die Punkte beziehen, die sich auf die innere Grenze der Oberflichenschichte beziehen, da aber 4 an dieser Grenze verschwinden, und mithin auch x, so folgt 00 == 0 da dieses in jedem beliebig liegenden Elementarprisma stattfindet, so muss dies auch dann stattfinden, wenn die Flachenintegrale tiber die ganze Ausdehnung des Korpers ausgedehnt wird. Mithin ne} =O iiberall, in so fern man von den Vorgiingen in der dusseren Oberflachen- schichte absehen kann. Das dritte Integrale ¢R wandeln wir auf dieselbe Weise um, in dem wir setzen on 2,0u of By. ee es ee Ox Ox so folgt is (2) cos 2x +2, cos ny + Qi, {213 COS us) Ou i ile -, +(@& cos 2x + 2, cos ny-+ Qa, Cos 23) ov aL (& f23, COS 24 ae f2sn COS MY “ee 23 COS ns) iw O uw LQ —\e dt Pee Soe Ge Pi Cust = Vin aaa Os a = CH Be Qo, Opt Qog\ x ‘i ieee is fos OEe,,...Op oH Vie r +( or op os f° Ou Ov Wo in 2, 2, etc in Allgemeinen die Factoren von oe Oy etc andere Werthe haben werden als in 2,, 2), etc da die Gréssen a, a, etc im Jnneren In wie fern kann man das Holz ete. 17 constant sind, aber an der Oberflache im allgemeinen Variabel sein miissen. Wir zerlegen das Raumintegrale in zwei, das eine ausgedehnt iiber die Oberflachenhiille innerhalb deren a, a, etc variabel sind. Und das andere tiber den iibrig bleibenden Raumtheil. In dem ersten Integral setzen wir wieder at=dwdn und ap 2, ap eae Op & OM Bry Op 2 = iI (23 ce asa ee es aa Oy am Tel etc da nach der eingefiihrten Annahme jeder Punkt des aus der Grenzhiille ausgeschnittenen Elementarprismas als starr betrachtet werden kann und die Gréssen a, a, etc von z allein abhangig sein sollen. Das in der Rede stehende Integral, wird, 0” cos zx etc von w etc nicht abhingig sind, a (2 cos 2x + 2, cos ny + 2), cos 22) Jou +( (2) cos nx + 2, cos ny + Bo, Cos 722) av +( (23 cos va + 23 cos ny + Q33 cos 22) ore ——— SL KG: 1 cos 74+, cos ny + 2.5 cos 23) jew — lw Aa 1 cos 7xr+2,, cos ny + 213 Cos 22) ype sim + Ee. (Qy cos 74a + By cos ny + Gy Cos 722) jew durch die Einsetzung dieses in @R heben sich dic Integrale auf, in denen 2, By etc treten so dass ok = — {to Le Non cos vx+2,, cos ny + 2), cos ns) au +((2 &, cos nx+ 2, cos ny + 23, cos nz) )dv +((L cos vr+2., cos ny+ 23, cos nz) Jaw 18 D. Kitao: af { ea & Oy Op 2» | Ou Pe) r oy Os U Op Oa Op Oe Op 2» x ( Ox Oy “hr ae =) +(4 fu Se + Bee Vaeu Wir erhalten als Gleichgewichtsbedingung \e ou+H dv+Z ow)+0R=0 da an der Grenehiille (on = Ou Ov = dv ow = 0w) Wir haben somit als die Bedingung fiir die Gleichgewichtslage 2, cos nxt+p 2) cos ny +p 23 cos 23) =E pe 2, cos nx+p 2» cos ny+ pp 23 cos uz)=Hh we Qy cos uxt p23, cos ny +p 23; cos uz) =Z Op Qu ua On =i Op 2s Qyy Ox Oy Wee Op 2. 4 ot Ba Opt Qo , SHE Coen OF Oy 2" am Op 2s Fe Op Lp Qo» re jee Qo, OF 8 © Oy Tec Vergleicht man mit der Gleichgewichtsbedingung OXx OXy. oma Ox Oy Tepe ove OYy Oye pa Oe 6 iy Vesa (eVises OZy » 3eZe Ae ae Gy... ea ae so erhalt man unmittelbar Xe=pQy, Xy=pQy, Xe=p Qs Ve=u2, Yy=p2, Ye=pQy Ze= pn Qy Zy=p Ou Ze=H Lx " Ou ov ow v ww Ow . Ou Xx= Ka, ra + ay “Oy a + anil ae, +) + a( Ox re In wie fern kann man das Holz ete. aw 2 Ky = (a seta 5 tas ge tals +5) +a oo +5e Ou Ov Ow TD Ow \.. WwW . Ou Xs=y (as A= tana Oy + aa ae +a,( 5 a 5) + an met 5S VES =H(a — + a; =. + asta ar Ow ie) Ow ¥y = aw $a, 3 + aS +45(5- ie = Yz= = ee Ey syaaias +a;~— = tay( 52 OV " ow )+a(2 2a wee Ou ow Oo Ze — rans tae ge + @; oa a5 (2 + \+a(Zo+ =) die 15 Coefficienten a sind die sogenannten elastischen Coefficienten, und sind Constanten ausgenommen die Punkte der unendlich diinnen Ober- flachenschichte. Diese Functionen lassen sich anderes schreiben, in dem wir itiberall einfihren pee, OU eure et ov! Som Ow Ou Emon Woy. 91Ss5 "oe som hor os" Ou , Ov oy oz s NX, = (aq, %2+ U2 Vy + Qy Set hg Sy + Ay Fz+ a5 V2) 20 D. Kitao: Y= (G12 Lp + Ae Vy + Gig 22+ As Pet Ms 12+ 4; Iz) Z,= [HQy Het Qo Vy t 43 22+ Ag Iz TU 4,+a5 Iz) Ly = Y= 3+ QsJy t+ As et Ay Sy t+ Us Vz + Qi Iz) X= Z,,= (dg Hp t+ Qu Vy tA Set Ay Fy t+ Qn 42+ M3 Iz) V, =X, = uy Xe t+ Ap y+ Os Sz t Aig Sy + Ay 12+ Me Yn) Man bilde PH4(Cyrd + 2C pte yt 2 sete t 2Cptn¥2t+ 2C or 2IV x + Cony? + 2C x: PyS2+ 2 Cu Vy¥n + 2Co5yV2 + 2Co6IyI'x 4+ CoS 74+ 2C Sey + 2CyyF%et 2C Fy) x == CuZy + 2Ca52yte+ 2 CugSy Vx + C5547 + 2C spt Ix + CoHn und differentirt man nach den 6 Argumenten Dae epee ee Or. Or. A Cue + Cpyy+ C2, 4+ Cysy+ Cy 54 ve+ Cure) evZ Z) =(Cx a, et Copyt Cuz. + CaSyt+ Cott Costa) Vy oP =a =(Ci y at Cog Vy + Co322+ Cut oy + Cysts + Card's) Y a a =(Ci 014 Oxon Gain eens y+ Cute) OF Or. (abated Cony + CysZzt Cassy + CoA ‘eat Cue) oF : OV, = (Cur ae CosVy + C32 + Cae, + Crt 4- Cos yz) Wenn wir hierin setzen =O MSC lh CS Q2= Co as= Cr as= Cx as= Cog As=Cx, Ay=O>s ay=Cy=Cy A= Cru=Cis Ay = Cy3= Cos Ay=2Cg— =Cis A= C=C A15= Cog= Cas so folgt Ob Sn a enn ef cs | ee Ae Hae yi ae Baa. Peet he Da, —— YS - In wie fern kann man das Holz ete. 21 7 oF L week Je) ss, aa BS ye ei, Be 2 ‘ oP } mee aia r es d. h. die Componenten der Druckkrafte haben also ein Potential. Die 15 Constanten lassen sich im Allgemeinen theoretisch auf eine geringere Anzahl nicht zuriickfiihren. Sie lassen sich, wohl, wenn der KOrper eine gewisse Symmetrie in seiner Struktur besitzt d. h. ein Krystall ist, der nicht dem triklinischen System angehort. Wir nehmen zunachst an, die Struktur des K6rpers sei symmetrisch in Bezug auf eine Ebene, nehmen wir diese zu (x 7) Ebene so verschwinden von dem Constanten a diejenigen in denen y in ungerader Potenz, vor- kommen, wie S a= {an F (0) $y So ist in diesem , i Ae We Ont =O Gg =O ils =O: | > ise Mithin Cy=0 Cu Cy Ge Cix=0 Cu —0Ca—0 (Ceo die Funktion P wird in diesen Fall = Oete, 1 20 at Py t 20 ats, + 2C 2%, + Cp PA 2C x VyS2+ 2Ca5%yS- + C37 +2C 522%, 2 Ci. yey Cig5 wx + Coste + Coy a" In der That bleibt diese Funktion unverindert. Wenn man hierin statt yu —y—v setzt, denn es werden ee of ne, OW = -( Ov , Ov ary os a) Aaya ae “Oy die Anzahl der Constanten reducirt sich auf 9. Ist der Kérper noch um eine Ebene etwa ys Ebene symmstrisch gebaut, so verschwinden alle von den a in denen + in ungerader Potenz auftritt. Es werden noch 4g = Az = Ay = Q4=0 Cas= C= C= Cu = C=O 22 . D. Kitao: die Funktion P wird in diesem Fall PAC yas £2 ets Vg Cte + Coy +2023 Iy5: + C37 + Cys, Ce + Cox’) W obei Cyx=Cag Cp=Ce eae Mithin P=3(Cy t,t Copy +. C52 + Cys(245%24 4) + Cy(24, Jy +z ) + Cy3(2 722+ 2,7) Die Anzahl der Constanten so ist 6. Die ry Ebene ist zugleich auch eine Symmstriebene, dann P bleibt unverandert wenn man ¢ mit —z vertauscht. Ein solcher Kérper gehért dem rhombischen System. Ist der K6rper so beschaffen dass die Struktur um eine Axe symmstrisch ist dass die Druck- componenten sich nicht anderen wenn man z. B. 2 mit y vertauscht so wird die Anzahl der Constanten noch geringer. Sei der Kérper symmetrisch um z-Achse, so ist dann a= Az a= \an'F(p) P= fan’ Foy! [dm # (o)ee9t = [dm Fe? Cy=C,° +Cg= C= Ga] G4. Mithin ist in-diesem Fall P=3(Culte’ +I) + Cog + Cyl 242Iyt+Iz) + Cys( 2492. + 27 y22+4)) Was sich in der That nicht andert, wenn x mit y vertauscht. Solche Kérper sind Krystalle des quadratischen Systems. Ist der Korper so gebaut, dass er zugleich zs. 2. umdie y-Axe symme- trisch gebaut ist, dass man auch z mit + vertauschen kann so muss Y=az a2 40 fame (aye = [etn Fp) In wie fern Kann man das Holz ete. 23 jan! (7 = ant Faery Ca Ga WOR Die Funktione P wird P=C)(4¢ +y, +22)+ C( 4,7 +97 + 2,7) +20 (4eVyt ZV y+ 22% x) Solche K6rper heissen regulére Krystalle. Die Druckcomponenten des, reeuléren Krystalls sind Ap =U Cutet CrIx+ Cr272) Yy=p2(Ciipy + Col, + 22)) Z, =O 52+ Col Iy + 22)) A= pC pry ¥o= pC 372 LG NS ONE Wir betrachten jetzt Kérper, welche dem hexagonalen System ange- h6ren, die Structur des KGrpers soll so beschaffen sein, dass sie sym- metrisch ist in Bezug auf drei auf einander senkrechten Ebenen, dass eine Drehung um 60° oder einen beliebigen Winkel in einen von der urspriinglichen nicht unterscheidbaren Stellung ftihrt. Es ist im diesen Fall P=4(Cyte + Copy $+ CapZet13(242Sn%2)+ C(24%2Vy I") + Co9(27yS'p + Zy)) die Hauptachse sei s-Achse, und in Holz wiirde s-Axe die Richtung des Longitudinalen sein. Wir fihren ein zweites Coordinatensystem 2’y' ein welche gegen das alte x y um den Winkel 7 geneigt ist 2=2 6—7 8 A@=Cos y u=s'S-+y 2 VPSsin x a? + (P=1 und daher u=u'a—v'8 v=ust+v'a Es ist 24 D. Kitao: du aay) ee dey Aik! ap N ee on dy! ed ir aw) pains : (ie +) ata tae) da da \ ef aa ae (= au! ( at aay ay i + ay )a- ay ES ar) oder wir die leichtverstandlichen Bezeichnungen einfiihren 4tu=t_ +4, —J-, Op Vr= HU! FP yy C+ y_' 48 In=(%_—Iy' OB +7q'(@ — F) 2,= 5,014, 4,=(4e—250 Wir bilden die Ausdriicke z aP. ae / ayy Aga Y= ‘P ae und setzen die Werthe der Druckcomponenten in neuen Coordinaten system . Ag Vy £2) 2y Veg ee aP ae aP See AZ eh AY =i aes Ze =H oa aP aE 2 aP Fela ie rey pm Rc Ti, Ve'= rs Pie nits: rT Ky =? ae Es ist nun Vita ,{ 22. a2, aP dy , dP ds, dP dy, GP dy 4 =I\ de dz’, dy dt',| dz, at', | dy, at',' By at di a O: 2D _ mp Was ee oe: De _ Bbq _ a: ay’. ay» asin dee Oe _ ay A Ce ee Aas ie Xa! + V,/?+2Y,a3 und auf dieselbe Weise Y',=X,7+),@—2Y 8 pate - In wie fern Kann man das Holz ete. to Ut ¥, =¥,=(¥%j—X)ap+X(e-F) Vij=2'=-X p+ Va Z,=X',= ¥n8+ Y, Es ist nun X= (Cy tet+ Coty + C32. = p(a'(@Cy + Cp’) + e2(48(Cy — Cy.) + Cilz,)) Y= (Copy t+ Crete + Ci332) $y", (Cu P+ Cre) = U4" 2 (Cj? + C20”) yy! (Cn? + Ch") + 9'x43( C22 Cy) + C2322) Agsp( Corts + Cigtz+ Cozy) = p42! (Cyy@? + C593") +9" ( Cor? + Cy?) +y'ya3(Ca— Cis) + C3522) Xy= Ve= pC wy2= pC 2(e'sy'y)a3-+y'e—P)) Ve ZC = pC (2 2 4- % 2) A= B06. eC (2 t= 2! fP) Die Einfihrung dieser Ausdriicke in (I) i a3 =2',(aCy + BC+ 2C PF + 407 C\o) +e, (22'(Cn + Ca) + C),(a* + Bt) — 4 C07 3( + 22 (Cis(a)* + Cos”) +x — (Cy — Cr) + /F(C2— Chr) + 2(a?— C2) a8 io ee rag =4',(0°F(Cy + Cx) + Cn(a+ 8) + 4024784) + ely (PC + a°Co5) + 3'( C433? + C28a°) + e!x( — '(Cu— Cr) + @(C— Cy) — 2(4 — 8) Cy) a8 Z'2 : f ; > aoe : 2 = (C430? + Cy3)°) +.'y( Cig 8? + Cosa”) + a8 7'2( Cos — Cis) + C322) Fag ™~ 9 9 ae — aj3x' .( Ce a Ca) a Biol G 93h + C56) A's = 4! 03( CF +2? Cy3 + Cy,(@ —%) — 5'y4;3(Co3— Cis) ry ae = 1',(48)(Cm? — a®Cy + Cy,(a — 9°) + 2C),(2— §*)) Wenn der Korper so gebaut ist, dass die Constanten von x unabhiangig wird wenn man die Coordinatenaxen um 7 dreht, so wird bei der Drehung um ¥ wieder sein 26 D. Kitzo:; x! = = ee at Cy’ poe Cy32" ‘ Y'y ; ' = Cry", t Ct’ + CyZ'e ZZ ; na: = Caen Cry yt Cpt XG! pe ys) NZ 7 2, 2 — ee So miissen sein AC, EC yh OCu er = Cr @B(C, + Cn) Cu(e + 2) —4C eo Cpe + C,.0' = Cg B(Cx2— Cy) =2(Cy— Cy.) +¢(@ — 2") Cr2=0 BC+ a'Cn) + 6Cy20'2’ = Cre C13 + Cy = Cis 2(Cyt Cy) +a(C, w— Cy) —2(@—-2)Cp=o Ce—Ca=0 II Cy? — PCy — C(—)2C),(@ — 2) =0 @F(Cy— Cy) +e —P(Cu— Cu) + Cn(o*— B= Crp diese 11 Gleichungen erfiillt man durch die Annahme Cis= G55) (Gu — Ome CoB? — Cy + C,(2—) +202 —P)=0 hi und da C+ = I ist. Mithin erhalten wir fiir einen K6rper, der um die s-Achse symmetrisch gebaut ist, wie bei den Hélzern, P=3C\3( 27 +9) + Costa + Sy? + 2422+ 2ySz) + Cog5z" = Cy,(242y 7] +42) diese Gleichung gilt auch fiir jeden Krystall, der dem hexagonalen System gehért, denn die Werthe y sind die einzigen durch die Gleichungen cleichzeitig erfiillt werden denn die letzten Gleichungen von I und II lassen sich auch so schreiben PC. —-€Cy + 3(@—-/)Cp=o PCy — (PCy + 3(A—)Cyp=0 In wie fern kann man das Holz ete. 27 Addision giebt denn GC Aus den beiden Gleichungen in III folgt Ca Ce S38 =| am! F(p) Sta, = [ant Foy — a 3) Fo) 7 Wir fassen jetzt einen Kérper ins Auge, welcher so gebaut ist, dass die Druckcomponenten sich nicht andern, wie man auch die Coordinaten- axen legen mag, einen Korper von dem man sagt er sei isotrop. Die Funktion ? behalt immer dieselbe Gestalt, wie man auch nie Coordinaten- axen drehen mag, als wenn der Ko6rper in jeder Richtung sich verhalt, wie ein regularer K6rper. Wir setzen daher P=Cylr2 +93 +52) + Cuoltp tye ee +20 3( te Vy+ Sey + 22x) und setzen zugleich Cy=—A(1+2' +h 2C y= — 2k A. Wo Kkk' gewisse Constanten sind, wenngleich nach dieses Definition k'=}3 sein sollte so lassen wir sie einfach bestehen aus Griinden, die wir spater noch unten auseinander setzen wollen. Es ist CytCe.=—k'K-K da K=—20Cy CntO,=—-K+2C, Cnu— Cyp=—hK —h'K= [am Fp) (F— 7°)F fir einen isotropen Kérper ist #’=0. Wenn also 2’ verschwindet, so ist der KOrper isotrop da in jedem um s-Achse symmtisch gebauten Kérper Q, = 32 ees af arate ) 28 D. Kitao: muss durch €y zum verschwinden gebracht werden, damit der K6rper als isotrop betrachtet werden kann. Die ¢ sind intermoleculare Grésse, und der Querschnitt des Holzstaibchen muss darum so klein werden, dass af pd +27F(p) verschwindet. Legt man die s-Achse so in die Lange eines K6rpers etwa des Holzes, und + und y-Achse so verringert, dass "i =e ape) fa en L()(S— 4°) verschwindend klein betrachtet werden kann. Ein Streifen Holzfaser ein Holzstaibchen von verschwindendem Querschnitt kann daher als ein isotroper Kérper betrachtet werden, die Funktion P wird P=-K(x, typ+s2tHeptyit sd) thaetyt2 $ext tye +32) die Gréssen bade ae te tip ae +4 (ty tye + 0° Ou Ov Ow Oi eye ee ou ov \? ow \? Ov ou \3 Ou , Ow (az) +a) +S) +354) tHe te) ow ow jit SCeee eee +3 oy + Og ) anderen sich nicht, wenn das Coordinatensystem gerandert wird, wohl ‘ p " Fo ae ov \? ow \* Mig oe rp + i (+) + (=) oi a um dies zu beweisen fithren wir die Hauptdilatation 4, 4, A, ein und bezeich- aber die Grdésse nen die Cosinusse dieser Richtungen, welche sie mit den Coordinaten schlicssen mit AAT Or Pai’ O27’ I's sind uns bekannt a Cu 95 6 ” 9979 ay =t,—= aga? ++ Che + ache O. ~~ —=—— In wie fern kann man das Holz ete. 29 OV aye AOS - ATs cea eer BEA ahs + Fath: ae ee =), + as) ,+ 07 ow Ou oe : ne : 3( Sa ay a a =P MAy + 2G2h2+ 732343 (Ss + a UGB ae Wenn wir jetzt die drei ersten Gleichungen addiren, so kommt sete cy ==: Le tIyt 22=Mthtss was die Pe eta, bedeutet und von den Coordinationsystem ganz und gar unabhangig ist Man findet ferner Ay typ tee Hai (art Bi +r) + 2A A,(aya? + By)? + 77773") + A?(a5+ B+ 72") + 2h A3(a7as' + BP 8s +7743) HA (a3! + 33 +73") + 2A A(as'as' + 3233 +72'7s') 2(2y +4,’ +72) = 2A (Br +r ar t+ %'B,’) + 2d? (Bry? + pay? + ay'3,") +243 (3373' + 73'as + 25 3s) HAMA Br F2F2 + 1272 + %/3122/32) + 4A 43 ((171 Fis 1718373 + 21/3332) + 4hohsl3u72/3si's t+ 42727373 + 22/3/95) die letzten drei Glieder lassen sich anderes schreiben. Es ist 4,4, + Ai 3.+ Hre=e das Quadrat hiervon ist > » afust 3 Be + wee +2(a, Oy 8:22 + QA 7'2+ Aipy iy2 ‘s) — (42a? + BBP +7772) = 2(a, 72,22 + A) oy ] yt Heyy 4 We ») 30 D. Kitao: ganz auf dieselbe weise — (afas' + B)33' +775) = 2063333 + HOt s+ Aussi 7s) — (aay + B83’ +7272") = 2 (5393 + O47 173+ 2233773) Er folgt hieraus Be +4) + 42) =24 (Br 7re +7er + ay) + 2A,(9)72' + 72@2" + 02/32") + 225(Bs73° +7343 + 43°23’) —24,1,(4/707 + B28 * +7772") — 2h A;(ayas + BBS +7173) — 2h, A(aya3' + BY 3s Frere die Addition dieses zu r?+y,?+27, so heben sich die mit 4,A, 4A, und 4,4, multiplicirten Glieder auf. Mithin Lg t+ Py +2, 4+3(2y +Ie +42) =MP(al + BY +77) +4M(al +BY +72) +A?(ay + B2+7%) =A) +A2+ A? d. h. unabhangig von der Lage der Coordinatenaxen, in dem fir P angestellten Ausdruck ist aber nach das Glied Bi! (ag +9, +22) vorhanden, das im allgemeinen von der Richtung der Coordinatenaxen abhingig ist. Soll aber Pin jedem rechtwinkeligen System unverindert bleiben, so muss R! verschwindend sein, d. h. irgend wie [am Hine —p)P=KK unendlich klein sein, damit der Kérper ein Isotroper sein kénnte, welchem System er auch angehéren michte. Wir erhalten fiir einen isotropen Kérper P=—K(aety teeth git 26 te) + hte tty t+2)2) Cy=—k(K+h) a In wie fern kann man das Holz ete. Die Constant & ist theoretisch in der That — we Es war fur jeden um die z-Axe symmetrischen K6rper Cu=3Cr was auch der Fall fiir den isotropen Korper ist = = = [ted(0) R ae Ee ST Sadia ange =3 nde Fp) Wir fiihren neue_Coordinaten €’7/é’ ein welche so definirt werden é=ael + 7! +78" 9 = 48! + Poy! + 725" i fo 4 a) $= a6! + 39! +735 wo a # 7 wieder die Cosinusse sind. Er ist fF G+ P= p=E" +7? +E" Cy =a pae(ak + Au tne) Fo) i=. — 2 Tas’) +7 +n ) (2) 3(a¢+ Pi +7) [cies *F(p) + 3(a"PY +BY : jiae)| nd re'y"°F(9) Da in Folge der Gleichmiassigkeit der Struktur des Kérpers |udeFp)32= [nae hp )8C"= etc =a) |utero) aa |uceFo)e7 =—eiC=0 juaeFo)e = [ya Fo)y" —Clo— mithin haben wir auch = (4+ 8 t+7)at sar srt arn't ra/)ay 2 Es ist nun T=(a7 + BP +7) =a +B tris 2(a78? + 877 tier) folglich a,=(4 +3ftri‘jat 3a2( 1 — ay’ — 3f—y') 32 D. Kitao: a,(1 - a; — 3 — 74) = 34n(1 — 2; — pi —7') folglich Oy = 322 mithin Cy=3C Ka +H=2# k=3 ob ein solches Verhiltniss herrscht ist eben so zweifelhaft wie der Einwurf dagegen, da jede kleine Heterogeitit des Materials stark dieses Ver- hiltniss beeinfliissen muss, und man bei jedem Versuche nie sicher sein kann dass, das verwendete Material homogen sei. Caginard de la Tour (Poggendorff Band 12S. 518) fand in der That fiir einen Eisendraht F, Naumann fand allerding fir Eisen den Werth £=4 und fir andere Stoffe nahrte sich & nach seinen Versuche dem Wertheimschen Werth. Die Untersuchung von Kirchhoff (Poggendorff Band 108 Seite 369.) haben das negative Resultat geliefert, dass zwischen den beiden Constanten fir die isotropen Kéoérper kein constantes Verhiltniss stattfinde. Cornu (Comptes Rendus T. 69. p. 333. 1899) fand tir Glas durch optisches Mittel, das Verhialtniss cna) ( 2k+1 zwischen 0.22 und 0.26, was den theoretischen Werth {=0.25 nahekommt. Mallock (Proc. Rog. Loc. V 29.157, 1879) fand fiir einen weichen Stahl das Verhialtniss 0.259 aber fiir andere Stoffe gréssere Werthe. W, Voigt (Wiedemann 15. p. 497, 1882) fand fiir ein galvanisch niedergeschlagenes Kupfer genau den Werth 1/4. jedoch kann man diese Abweichung keines- weges der Theorie zu Grund liegenden Hypothese zur Last legen. Denn tole die Frage ist eben noch eine offene, in wie weit die beobachteten Abwei- chungen der Ungleichmissigkeit des Gefiiges des Materials zur Last fallt. Es mag demk sein, wie es sein wolle. Es handelt sich darum, das In wie fern kann man das Holz ete. 33 Holz so zu dimensioniren dass man es als isotrop betrachten kann, dass |ndeeF(o) als unendlich klein ansehen kann. Ich habe vierekige Stabchen von Holz bilden lassen von ungefahr 1 cm Quadrat und von der Linge 60 cm nnd dieselben der Biegung unterworfen, und K auf gewdhnliche Weise bestimmt. Unterscheidet dabei eine Biegung in tangentialer Richtung, und eine Bieguug dazu senkrechter radialer Richtung, uud nahm an, dass die beiden Biegungen dieselbe Grésse K liefern miissen, wenn das Holz als isotrop betrachtet werden kann. Das Instrument, das ich angewendet habe, war wie folgt, beschaffen (Fig 1). Eine etarke eiserne Bank DD tragt zwei scharfe Keile LL, die man langs einer Rinne in der Bank auf und niedergeschoben werden kann. LL dient dem Holzstabchen HH als Stiitzen, CC ist eim Metal- rahmen mit einem Keil, und hat eine Spalte, so dass man den Keil genau auf die Mitte des Holstabchens !egen kann. Ein Spiegel! von grosser Brennweite dreht sich auf dem Stifte T, und ruht auf dem Stiftchen S, welches das Plattchen M (Fig 2) beriihrt, das Plattchen M ruht auf einen Biigel CC, der mittelst der Schraube 0 amdem Metallrahmen AA auf und niedergeschoben werden kann. Die Schneiden LL kédnnen mittelst der Schrauben s in jeder Hohe befestigt werden. Der Metallrahmen (AA) steht mit dem Metallrahmen B in Verbindung, und das Holzstabchen H ruht auf dem Keil &, und wird durch ein Gewickt G in der Schale herabgezogen, und so gebogen, in dem das Holzstabchen H gebogen wird, wird S herabgezogen und so wird der Spiegel B gedreht, und das Spaltenbild n wird emporgehoben bis m. Der Winkel i um den der Spiegel gedreht wird gemessen und tagz =tag 2a Pp Ey ad SA OTNS ges der Biegungspfeil 0 ist dann annahrend é=/. sin @ Wo 7 die Lange des Stiftchens (S) ist. Indem 0 auf und nieder schraubt kann m mit n zum Zusammenfall gebracht werden. Die 34 D. Kitao: Empfindlichkeit des Instrumentes ist dann sehr gross, und ein 1/1000 Milimmeter von dem Biegungpfeile kann leicht bestimmt werden dass gungp manche Ubelstande sich zeigen, wie der Einfluss der Eindriicke an der Schneide, die den Holzstiben als Stiitzen dienen, der Einfluss der Luftfeuch- tigkeit. Breite des K. K. Standort Altr Fallzeit Herbstrings} gebogen tang | gebogen radial} Location Thuya obtusa, (Hinoki) Takaosan 130 Marz 0,140cm | 14,03 x 107 12,93 X 107 Kern = = = 0,100 cm 13,73 X 107 13,15 X 107 < Gifu 160 unbekannt 0,030 7,13 X 107 6°72 x 107 * Kishu 130 | November 0,150 11,70 X 107 11,79 X 107 % a Hy | a 0,170 11,70 X 107 11,70 x 107 = 2A 60 | August 0,140 14,48 x 107 14,80 x 107 on 3 “ | os 0,150 14,37 X 107 15,27 X 107 Pr Takaosan 130 Marz 0,150 14,86 x 107 12,88 x 107 |} Kern Splint » ” ” 0,050 14,89 x 107 13,58 X 107 Splint Gifu 160 unbekannt 0,060 14,26 x 107 12,89 x 107 . ” ” 9 0,090 6,56 x 107 6,25 x 107 + Thuya pisifera, (Sawara) Takaosan go December 0,090 11,16 Xx 107 11,41 X 107 Kern a e PP 0,110 10,70 X 107 11,28 x Io7 Ss ” » ” 0,130 10,75 X 107 10,99 x 107 ” ” ” ” 0,080 ” ” Splint ” ” ” 0,080 11,18 x 107 10,16 x 107 4 59 ” * 0,090 9,69 x 107 8,70 x 107 ” In wie fern kann man das Holz ete. Breite des K. Kx. Standort Alter Fallzeit erbstrings| gebogen tang | gebogen radia!| Location Thujopsis dolabrata, (E1Liba) Kiso 150 unbekannt | 1,190 10,38 x 107 10,35 x 107 Kern ” ” » 0,260 10,93 X 107 11,61 x 107 45 ” ” 9 0,200 8,68 x 107 8,69 x 107 |4 Kern 3 Splint ” » » 0,170 9.81 x 107 9,36 x 107 Splint Sciadopytis verticillata, (KGyamaki) Kiso 80 November 0,30 6,55 x 107 6,06 x 107 Kern | unbekannt | unbekannt|] unbekannt 0,200 10,81 X 107 10,87 x 107 | 55 Kiso 80 November < 6,58 x 107 6,43 x 107 Splint unbekannt unbekannt | unbekannt 3 11,16 X 107 11,39 X 107 *y Cryptomeria japonica. (Sugi) | Takaosan 200 November 0,095 sss Se OV Seg) Kern 0 9 “” 0,070 5,17 X 107 4,98x 107 | » | Kisha 140 unbekannt | 0,120 6,35 X 107 604x107 |t Kern ¢ Splint | | | . ” ” | ” | 0,130 5,19 xX 107 5-64x Io” 3 Kern 3 Splint | | | : Tsuga Sieboldii. (Ssuga) s Boe 7 2 — Kiso 100 November 1,180cm | 11,35 x 107 11,5Q9x 107 | Kern ” ” ” 0,210 ly -Osssnerer 5,64 x 107 Sj lint | | | t 36 D. Kitao: Breite des K, 1K Standort Alter Fallzeit Herbstrings}gebogen angent|] gebogen radial} Location J = Zelkowa accuminata. (Keyaki) Takaosan unbekannt | December 0,112 9,41 X 107 9,98 x 107 Kern a3 a » 0,158 10,11 x 107 10,77 +107 sf » 9 » 0,165 10,09 x 107 10,46 x 107 " » » ” 0,120 9,54 x Io7 9,95 X 107 Splint % 9 0,620 4,69 x 107 5:93 x 107 5 » : ” 0,650 4,53 x 107 4,57 x 107 % Castanea vulgaris var. Japanica. (Kuri) Takaosan 40 unbekannt | 0,126 13,21 x 107 13,36 X 107 Kern ” ” - 0,290 12,00 x 107 12,38 x 107 a ” » 39 0,280 12,03 X 107 13,21 x 107 Splint : cf 5 0,300 13,07 X 107 15,25 X 107 » Magnolia hypoleuca, (H6) Takaosan | unbekannt | December 0,230 9,94 x 107 11,36 x 107 Kern » 9 y 0,250 10,62 X 107 11,04 X 107 _ | 9 ” re, 0,190 10,22 X 107 11,42 xX 107 a ” ” a 0,300 10,15 X 107 11,01 X 107 Splint ” ” 3 0,165 9,85 x 107 10,83 x 107 y Fagus sylvatica var. Sieboldi. (Buna) Takaosan December 0,130 12,16 x 107 13,52x 107 |} Kern 4 Splint ” | ve 7 0,160 11,41 X 107 12,22 x 107 Splint 11,31 X 107 12,96 x 107 In wie fern kann man das Holz ete. 37 Breite des K. K, Standort Alter Fallzeit dal tac ial tangent} gebogen radial} Location Quercus glauca, (Shirakashi) | Takaosan unbekannt | December 0,160 11,69 x Io7 | 11,71 X 107 | Kern | Quercus acuta. (Akagashi) unbekannt 40 unbekannt | 0,500 13,18 xX 107 13,36 x 107 Kern ms 40 By 0,350 PEATE SS LICOYS 12,68 x 107 Splint Quercus thalassica. (Matebagashi) | Nishigahara | 25 | September 0,45 12,10 X 107 | 12,19 X 107 | Kern | Fraxinus Sieboldiana, (Shioji) Chichibu 48 unbekannt 0,276 15,02 x Io7 14,73 X 107 | Kern » » or 0,200 14,13 X 107 14,41 X107 | 9 | | Panlownia tomentosa. (Kiri) See | Nishigahara 7 September 0,400 4,16 x Io7 520x107 | Kern » ” % 0,450 4,62 x 107 4,12 x 107 38 D. Kitao: Pinus densiflora. (Akamatsu) K, K. Standort Alter Fallzeit Breite des eebesen gebogen Location Herbstrings aoe ; aE tangential radial Mimuneya 95 November 0,123 10,36 X 107 9,03 X 107 Kern 2? ” ” 0,136 9,01 Xx 107 9,68 Xx 107 Splint Takaosan 50 - 0,180 11,86 X 107 11,52 X107 Kern FP 2 3 0,236 12,73 X 107 12211xX 10% Pa ” ” ” 0,350 10,94 X 107 T1,31-X10" Pa » ” 2 0,198 10,94 X 107 11,59 X 107 Splint ” ” 9 0,175 10,56 % 107 10,47 X 107 44 ” ” 5 0,197 10,73 X 107 10,08 X 107 Abies umbellata. (Momi) Takaosan 160 December 0,200 9,34 X 107 9,12 X 107 Kern 9 0» 99 0,120 9,33 X 107 9,86 X 107 : » 93 %» 0,230 10,48 X 107 10,73 X 107 rf ” ” % 0,170 10,26 X 107 10,14. 107 Splint ” ” » 0,260 9,67 X 107 9,89 X 107 2 ” 9 a 0,360 10.18 X 107 9,76 X 107 s Ich habe 6—7 Male Biegungspfeile bestimmt und K fur jeden Pfeil des Stibchens ermittelt und daraus das Mittel genommen. Wenngleich die Stabchen sich seit zwei Jahren in Zimmer des Laboratoriums befanden, und darum vollkommen lufttrocken sind, so war der Einfluss der Luftfeuch- tigkeit auf den Werth von K so gross, dass die erste Decimalstelle sich oft aindert, dass die Luftfeuchtigkeit mehr die Anderung des K verursacht als die Breite des Herbstrings. In sofern die beiden Werthe von K mit der Breite des Herbstrings in keinerlei Zusamriennang stehen, und ihre Differenzen nur durch die Luftfeuchtigkeit veranlisst sein kénnen, kénnen Tn wie fern kann man das Hole ete. 39 wir annehmen dass die beiden Werth von K einen und denselben Werth zeigen kénnten, wenn der Querschnitt des Staibchens viel geringer gewesen ware, als ein Quadrat 1cm. .Ich behalte mir vor, Stibchen von viel geringeren Querschnitt in Untersuchung zu nehmen. ' ‘ ’ ) : ~ s =e f . y z - ‘ ? » Wolly cig aris-us iret oer wt a > =f ee = r ST. sed mers Fi f F ‘ .@ l ’ bj 7h hind hte ven tke Severe Ste, ; oa a #t woe TERE * (Ay vr , k Cote sf ' i hf ci “= ° , ws Ts ‘ : a | re ar ' z é bd adteilos eatidas THU at Bah an Fa = ay ™ ‘ = n : | = a 7 AAS TR oes ePit ape me Uhr Bias 7 > pe ; F rau 4 ~ ; \ oe | - at 4 : eS ‘ zi 7 Pa @ ir e s ) » ~ i . ean : Piel ts a « z : o \ 4 ors? D < A i es eS i : 4 ¢ © ieee > 7 4 1 4 7 ? e Ce : ° A a ' ‘ a sé 4.4 v ; i “ ‘ ‘ * ei of . u wry a ‘ 7s ' vr i ; ” '. a ~ he TAFEL 1, ofay o bore a i Pie AGKIC. COM: VOL, V: TALES. LL. Studies in the Physiological Functions of Antipodals and the Phenomena of Fertilization in Liliaceae. I. Tricyrtis hirta. BY T. Ikeda. With Plates III—VI. Introductory. Recent investigations have brought to light many interesting phe- nomena relating to the reproduction of Angiosperms. We have an enormous number of papers upon this subject, which are indeed valuable on account of the important morphological data which they contain, but from the physiological point of view, there still remain many problems to be solved ; among others, we may cite the physiological functions of the so-called antipodal cells. In fact, the opinions of the authors respecting this problematic organ within the embryo-sac do not agree, and it is easy to see that this divergence of opinion is mainly due to the _ specific difference of the plants used by the respective investigators. A general conclusion from the phenomena relating to antipodals cannot therefore be drawn until researches on various and widely different types of plants have been made. The investigations of antipodals have till now been restricted to the Ranunculaceae, Leguminosae, Compositae, Gramineae and a few others ;! and since similar researches on the Liliaceae, which have been so often subjects of investigations in regard to the phenomena of fertilization, were still wanting, except some short remarks by Westermaier? on a few 1 Westermaier, 1890 and 1896 ; Osterwalder, 1898 ; Goldflus, 1898—g. 2 Westermaier, 1896. 42 T. Ikeda: forms (Muscart, Hyacinthus, Allium), 1 began to make a study of this group of plants in this respect, not however neglecting to study the phenomena of fertilization. First of all, I took up as the subject of my researches 7yicyrtts htrta Hook., native in this country. The results of the investigations discussed in this paper still present some gaps, but since, on account of other business, I cannot for a while continue work in this line, I will publish them here as I now have them. Materials and Methods. The material was gathered from September till the middle of October 1900, when the plants were in full blossom in our botanical garden. The methods of investigation were as follows :— 1. Free-hand sections of fresh specimens: besides observations on microtome-sections, microchemical reactions were tested on free-hand sections from fresh materials. 2. Microtome-sections: The material was immediately fixed after its collection and serial sections were made according to the ordinary methods. Flemming’s strong and weak solutions, absolute alcohol, and Keiser’s sublimate acetic acid mixture were employed as fixing media. Of these, Flemming’s weak solution was mostly used, but its action was somewhat inferior to the stronger one, which always afforded excellent results. After dehydration through ascending grades of alcohol, the material was put successively in xylol-alcohol and pure xylol, and then imbedded in paraffine through xylol-paraffine. The sections were cut 5—10p thick. For staining, several reagents were used: Flemming’s safranin-gentian- violet-orange, Delafield’s haematoxylin, haematoxylin-glycerine, Heiden- hain’s iron-alum haematoxylin, Schaffner’s anilin-safranin, his picro- nigrosin, Baumgarten’s acid-fuchsin and methylene-blue, Gram’s gentian- violet and fuchsin iodine-green mixture. Besides these, various combin- ations of stains were sometimes tricd. Of these, the first mentioned always proved to be the best, though the others, except Schaffner’s reagents, gave pretty good results. Studies in the Physiological Functions of Antipodals, ete. 43 Formation of the Embryo-sac. The formation of the archesporial cell has nothing extraordinary: it arises as usual at the expense of a sub-epidermal cell, which soon becomes conspicuous by its larger size. In its earliest stage, the archespore has the homogeneous cytoplasm, which is equally distributed therein. Its nucleus, large and spherical, has its single nucleolus suspended in the nuclear reticulum (Fig. 1). No tapetal cell is cut off, so that the archespore deveiopes directly into the embryo-sac-mother-cell. The amount of chromatin, which is rather scanty in the earliest period, soon increases until it forms a thick convoluted spireme thread. Often beautiful mitotic figures are visible in the nuclei of the nucellar cells, but any calculation of the number of the minute and crowded chromosomes was impossible. The embryo-sac-mother-cell grows into a somewhat funnel-shaped cell, of which the distal end intrudes within the underlying tissue (Fig. 2 and the following). During the growth of the embryo-sac-mother-cell, the chromatin of its nuclei arranges itself as usual in a fine, long, convoluted spireme thread. The nucleolus is still present in the nucleus, which ts surrounded by the dense cytoplasm (Fig. 2 and 3). The spireme thread gradually shortens itself, leaving a clear space around it. The cytoplasm, which was homogeneous and dense up to this period, becomes now more or less reticular in its structure, which is probably caused by the rapid growth of the cell (Fig. 4). The convoluted spireme then undergoes a longitudinal splitting (Fig. 4, 5), which is soon followed by its transverse division into chromosomes (Fig. 6), whose number is not exactly known. The microsomes which constitute each chromosome are often clearly visible. Of these, those which are found at both ends of the chromosome gradually enlarge and this expansion of microsome granules is always accompanied by the shortening process of the chromosome, until it be- comes condensed into a short dumb-bell-shaped rod and when the con- densation proceeds, the final products are nuciear tetrads (Fig. 7); besides, X-, V-, and ))- shaped chromosomes are also visible. These chromosomes, which now lie along the nuclear wall or are suspended on 44 T. Ikeda: the delicate fibres of the linin-reticulum, arrange themselves on the equatorial plane of the spindle; and then the axis of each chromosome runs parallel to that of the spindle (Fig. 8). Of achromatic figures, the conducting fibres alone are visible, and do not converge towards one point. Of the nuclear tetrads, six were calculated in many cases, and though the number of chromosomes in the nuclei of vegetative cells, for example those in the nucellar cells, could not be counted, yet it is clear that the number of chromosomes in the latter case must be far greater than in the case of the division in the embryo-sac-mother-cell ; besides, the mode of division, as before described, is the so-called heterotypic one. From these observations, as well as from a comparison with those of various plants, it is clear that the numerical reduction of chromosomes takes place in the embryo-sac-mother-cell. After the formation of the septum between two daughter cells by the first division (Fig. 10), their respective nuclei undergo a second division, which may take place at the same time or at times differing in the case of each of them. It must be noted that the daughter nuclei produced by the first mitosis never enter the resting stage and soon begin to undergo the next division. When the second mitosis proceeds, the minute colourless particles as well as the highly stained bodies are found scattered in the cytoplasm (Fig. 11). These bodies might be regarded as fragments of the nucleolus, which do not perform any function in the mitosis—the so-called extranu- clear nucleoli. Of the four sister cells produced by two successive divisions, the upper three obliterate, are gradually driven off in the micropylar direction on account of the active growth of the lowermost one, and at length are visible as cap-shaped pieces directly above the future embryo-sac (Fig. 12). Thus in the formation of the embryo-sac in TZvicyrt¢s hirta there takes place a tetrad-division! of the first type, as recently stated by Schniewind- Thies.2 Guignard,? in his researches on the embryo-sac formation, studied also 7ricyrt’s hirta ; according to him, only one, the lower of the 1 In the sense of Juel (cf. Juel, 1900). 2 Schniewind-Thies, 1gor. 3 Guignard, 1882. on Studies in the Physiological Functions of Antipodals, ete. AS two daughter cells formed by the first mitosis, undergoes the second, so that only three cells are formed and this mode of development corresponds to the second type of Schniewind-Thies. The divergence between Guignard’s results and my own may be explained on the ground that he perhaps overlooked the second division in the upper daughter cell, which might easily have occurred at the time of his investigation (1882), when only free-hand sections could be examined. Maturation of the Embryo-sac. The cell destined for the embryo-sac now begins to grow vigorously by the absorption of food materials through the underlying tissue. It enlarges immensely; its growth is not, however, accompanied by an increase of cytoplasm. Its chalazal end, which as before stated, had been prolonged into a funnel-shaped body, gradually penetrates into the under- lying tissue in the direction of the median longitudinal axis of the ovule, and serves probably as the haustorium for collecting the nutriment for the embryo-sac. In (this case the single nucleus, which is more or less flattened, lies near the neck of this haustorium ; as it seems to me probable that the nucleus tends to occupy the best position for the elaboration of food for further development, the position of the nucleus in this stage is the natural consequence of the constructive metabolism in this haustorial part (Fig. 12). The cytoplasm of the embryo-sac in this early stage is small in amount and consists only of a few delicate strands along the wall. Besides there are present a few granular particles, especially plentifully deposited around the nucleus (Pl. VI, fig. 43); they have been proved to be some kinds of dextrine about whose distribution in the embryo-sac and else- where the later chapter is to be consulted. The embryo-sac nucleus in this early stage is relatively deficient in chromatin, which is suspended on the linin-net-work, together with two or three spherical nucleoli (Fig. 12). After a certain period it undergoes the first division and the two resulting daughter nuclei gradually depart toward opposite poles, until at last they reach respectively the two extremities of the embryo-sac (Fig. 13). They are small in size, with a round nucleolus in the centre. At this stage the cytoplasm still remains 406 T. Ikeda: unincreased and attached simply to the periphery of the embryo-sac, except around these nuclei, in which dextrine granules are deposited. I have had no opportunity of observing the following two successive divisions of the embryo-sac nucleus, by which eight nuclei are formed and the bipolar grouping of the cells is completed. The eight nuclei produced by these processes soon become bipolarly grouped at the two opposite ends of the embryo-sac, and both groups of cells show at first no difference whatever from each other (Fig. 13). At the beginning of the bipolar grouping all these nuclei agree in their structures: their chromatin granules are small in amount and are suspend- ed on the achromatic fibres traversing the intranuclear space; they have each a single nucleolus, which sometimes contains a few vacuoles. The cytoplasm of the embryo-sac is finely granular and plentifully provided with fine colourless particles, which are proved by microchemical methods to be some kind of dextrine. The embryo-sac, which is still in rapid growth, possesses large vacuoles in its centre, so that the communication of the egg-apparatus with the antipodals is by means of cytoplasmic strands through the axial portion of the embryo-sac, in which the polar nuclei are suspended. The antipodal cells now fill up the chalazal protuberance of the embryo-sac (Fig. 14). Their cytoplasm becomes soon afterwards very granular and compact having no vacuoles, while on the other hand the egg-apparatus is. somewhat deficient in cytoplasmic contents and highly vacuolated. The nuclei of the antipodals are always larger than those of the egg-apparatus, because after the completion of the embryo-sac the former undergoes much more vigorous growth than the latter, until at length, before the time of pollination, the antipodal cells become several times larger than the egg-apparatus. The union of the polar nuclei also takes place in this stage (Fig. 15). These nuclei as well as their nucleoli are spherical and of immense size, and besides are furnished with some refractive vacuoles. When the two polar nuclei fuse together, the nucleoli of both usually unite at the same time ; but sometimes they remain separate long after the union of the nuclei, for I have sometimes met with the embryo-sac nucleus with two large nucleoli, even shortly before fertilization. Se ee -" ee es ee ee Studies in the Physiological Functions of Antipodals, ete. 47 The fusion-product of these polar nuclei, i.e. the primary endosperm nucleus, is characterized by its large size and spherical shape, as well as by possessing a large, highly stainable nucleolus (see for example fig. 16, p.); the amount of chromatin is much greater than that of either of its two components. The union of the two polar nuclei takes place in the upper half or near the centre of the embryo-sac, but the fusion-nucleus sinks towards the top of the antipodal cells and again takes its route upward when the egg-cell is ready for fertilization. The primary endosperm nucleus, which is at first spherical, gradually becomes elongated and irregular in shape. The amount of chromatin does not show any increment corresponding to its growth, but its single nucleolus (or often two nucleoli) becomes extremely large, and shortly before the entrance of the pollen- tube refractive vacuoles of various sizes are always found therein. Shortly before fertilization, the outline of the nucleus becomes very irregular, often taking the lenticular shape in optical section. Of the egg-apparatus, the largest is the ovum, which locates itself somewhat eccentrically along the lateral wall of the embryo-sac (Fig. 16). It is always vacuolated in its basal portion and furnished abundantly with certain reserve-materials in the cyto-reticulum. The two synergidae are somewhat smaller than the ovum and also are vacuolated ; they have no reserve food material and often persist to the time of the early endosperm formation, but sooner or later they undergo degeneration, usually at the time of fertilization, but sometimes afterwards. The increase of the cytoplasm of the embryo-sac goes on parallel with its vigorous growth in size, so that the whole space of the fully grown embryo-sac is filled with the homogeneous and compact cytoplasm. . This increase of cytoplasmic contents seems to begin after the union of the polar nuclei. Antipodals, Integuments, Nucellus, Chalaza, etc. a. 6 rE TS 4, Studies in the Physiological Functions of Antipodals, ete. 49 during their activity. Ofsecretory cells in the animal kingdom we have various interesting examples described in Rosenberg’s work on Drosera,} which will not be repeated here. Turning to vegetable secretory-cells, we have an extensive work on septal glands by Schniewind-Thies. Her conclusion is as follows :?—‘‘ The nuclei of secretion-tissue of nectar are always distinguished from those of parenchyma by their containing a large quantity of chromatin......... The abundance of cytoplasm and probably sometimes also the extraordinary size of nuclei in nectar cells stand in relation to their great activity, for they must absorb raw materials necessary for the nectar formation, elaborate the latter by themselves, and transport this nectar outwards............ * Huie*, and later Rosenberg*, observed the chromatin-aggregation in tentacle-cells of Drosera leaves when they are nourished with various substances. Also recently W. Magnus? discovered similar phenomena in digestive cells of the endophytic mycorrhiza of various Orchideae. In his study on Aconttum Napellus, Osterwalder observed the same phenomenon in the nucleus of antipodals and brought this into relation with their nutritive activity.* All these observations of various authors as to the significance of chromatin-aggre- gation, together with other concomitant circumstances, which are to be described later, led me to conclude as probable that the chromatin-aggre- gation in the nuclei of antipodals of Tricyrtis is also the expression of their metabolic acttvity,—that therefore these organs play a most essential role in the nutrition of the embryo-sac,—that they are indeed the metabolic centre Jor the absorption, elaboration, and transportation of nutritive materials of the latter. This nutritive function of antipodal cells seems to continue from the time of the full maturation of the embryo-sac till that of the endosperm formation. The antipodal cells change their structure again after fertilization, when assimilation and secretory activities are gradually weakened and approach their end: the big chromatin masses produced by the aggregation now begin to dissolve gradually, so that they become smaller and smaller and the single nucleolus, which has been found 1 Rosenberg, 1899. + Rosenberg, I.c. 2 Schniewind-Thies, 1897. 5 Magnus, Igoo. % Huie, 1897-1899. 8 Osterwalder, Lc. 50 T. Ikeda: ‘ throughout the former stages, degenerates. The antipodals sometimes elongate upwards along the wall of the embryo-sac and in such later stages their cytoplasm takes a very characteristic feature: it becomes fibrillar and imitates the structure of pancreas-cells in Amphibia, as described by Mathews;! the arrangement of these fibrillae is very varied and often they are more or less coiled up, somewhat similar to the case of Amphibia, studied by the same author (Fig. 19,20). According to him,? the pancreas-cell is filled before secretion with metaplasmic zymogen-gran- ules, which disappear during secretion, the cell meanwhile becoming filled with protoplasmic fibrils. The fact that at the end of their activity the antipodal cells become filled with fibrillae, is an interesting cytological feature imitating the behaviour of the pancreas-cells above described. The antipodal cells devoted to the nutrition of the embryo-sac throughout all the developmental stages are finally gradually reduced in size and driven off downwards by the growth of the endosperm; or sometimes, they are seen attached to the lateral wall of the embryo-sac, entirely surrounded by the mature endosperm. In the later period of endosperm-formation, they are more and more carried away downwards, till at last they dwindle away to small flattened pieces (Fig. 48) and finally disappear entirely. 6. Integuments, Funiculus, and Raphe. The outer and the inner integuments of the ovule are each composed of two layers of cells. The limiting membrane between the outer and the inner integument, as well as that between the latter and the nucellus, which is only one layer thick, are already cuticularized in the earliest stage, except in the micropylar region of the inner integument and the nucellar cap (Fig. 44 and the following). I have tested microtome-sections, made from materials fixed with sublimate-acetic acid mixture and absolute alcohol with chloriodide of zinc, and observed that these membranes are never coloured blue or violet, but always yellowish brown. They are insoluble both in concentrated sulphuric acid and in copper-oxide- ammonia. Through the whole stage of development, their reactions towards such reagents remain unaltered. These cuticularized membranes 1 cf. Wilson, 1900, p. 44. 2 Wilson, l.c., p. 350. Studies in the Physiological Functions of Antipodals, ete. 51 gradually increase in thickness. Therefore, from the earliest period there exists no direct communication between the integuments and the nucellus or the embryo-sac through the limiting membranes. But it is an interest- ing fact that throughout all stages the micropylar region of the inner integument never undergoes cuticularization ; the possible significance of this phenomenon will be discussed later. The funiculus and the raphe consist each of four structural elements, namely the epidermis, cortical parenchyma, phloem and xylem. In the early period of the embryo-sac development the procambium string composed of cells with much elongated nuclei is designed for the conducting of tissue of nutriment towards the embryo-sac. Subsequently its function is taken up by the vascular bundles, which are themselves developed from this procambium. In the cross-section of the ovary, which at the same time passes longitudinally through the median plane of the ovules (Fig. 21), we can clearly trace the course of the vascular system through the funiculus and raphe. This vascular bundle in the ovule is the branch of the main-trunk running through the ovary in its axial direction. Before entering into the funiculus the vascular system passes through a special group of cells, which is placed near the placenta (Fig. 21, p/., Fig. 22a and 4). These cells are characterized by their small size, relative- ly large nuclei and abundant cytoplasmic contents, and persist unchanged throughout the whole period. The xylem of this vascular system consists only of a bundle of spiral tracheides, while the phloem is formed by a number of long columnar cells with delicate membranes, forming two or three layers around the xylem. The presence of sieve-tubes is doubtful. The cortical parenchyma consists of elongated cells with large rod-like nuclei. These vascular bundles run through the funiculus and their termination is found among a special group of cells in the chalaza. This group of cells, (Fig. 21, ch.) similar to that near the placenta, is characterized by their constituents, which are small in size, rich in cytoplasmic contents and furnished with a round and relatively large nucleus rich in chromatin. Afterwards the nucleus of these cells becomes gradually small and 52 1. Ikeda: irregular in shape until the intranuclear space becomes apparently empty ; and then the cytoplasmic contents of the cells disappear and are replaced by a hyaline fluid. These cell-groups, in the chalaza as well as near the placenta, are probably the place of the enzyme formation, as will be shown later; the special character of the cells of these groups speaks also for their nature. c) Nucellus and Chalaza. When we examine the nucellus in its young stage, we find that its portion underlying the embryo-sac is composed of polygonal or cubical cells, which are arranged in the regular manner, forming only one layer around one axial cell-group. (See Fig. 23, which shows the cross-section of that region of the nucellus). At the beginning there is no visible difference among these cells: they are all characterized by the presence of a large amount of cytoplasm as well as of a huge, spherical nucleus rich in chromatin. Through the rapid growth of the embryo-sac in the direc- tion of the longitudinal axis of the ovule, that portion of the nucellus becomes more and more elongated and narrow. While the cells of the outer layer retain their original cubical shape, those of the axial group become gradually elongated, until each cell takes a long columnar shape (Fig. 17) and its nucleus becomes correspondingly long. This is due not only to the longitudinal growth of cells, but probably also to the pressure exerted upon them by the surrounding ones. This axial row of nucellar parenchyma has already reached its maximum elongation at the stage of the full maturation of the embryo-sac. It has various names given by many authors, for example ‘‘ Conducting passage ” (,,Zuleitungsbahn”), ‘Starch route” (,,Starkestrasse”) by Westermaier ;* ‘‘Pseudo-chalaza” by Goldflus,? etc. It persists even to the later period of the endosperm-formation, though in a more or less degenerated condition. Cells of the outer layers, which are characterized by their large nuclei, never become elongated like those of the axial row but undergo gradual decomposition at the time of the maturation of the embryo-sac. This degeneration begins with the decay of delicate, parenchyma cells directly 1 Westermaicer, I.c. 2 Goldflus, l.c. _ == «eset th Studies in the Physiological Functions of Antipodals, ete. ur Oo beneath the embryo-sac: at first their nuclei disorganize, then the cytoplasm and and lastly the cell membranes. In this process, when the nucleus begins to disorganize, it becomes highly stainable ; its chromatin granules become scattered within it; but no linin-network is visible and no nucleolus is met with. When the nuclear membrane fades away, the nuclear contents are scattered throughout the cytoplasm. It is very probable that this process of degeneration contributes in no small degree to the endosperm-formation. The products of degeneration may be directly absorbed by the antipodals or conveyed through the conducting passage to the antipodals, which elaborate these materials for the purpose of the endosperm-formation. When the embryo-sac is already fully matured, it has been often observed in preparations made from materials fixed in Flemming’s strong solution that an immense aggregation of granular substance is found in the cells of the conducting passage (Fig. 17). It is probable that these granules are the protein matter derived from the degeneration of the nucellar cells above described, precipitated by Flemming’s solution during the course of their transit through the conduct- ing passage. Itis also evident that certain soluble ferments must intervene for this degeneration of nucellar cells; where these enzymes are formed is not clear, but it is not improbable that they are formed in the antipodals themselves, for the destruction of nucellar cells begins close to the antipodals and proceeds gradually downwards. The portion of the nucellus, which forms the lateral side of the embryo-sac, is only one cell-layered. It begins to disorganize generally at the time of fertilization, but remains till a pretty advanced stage of the endosperm-formation, though in a degenerated condition; finally it be- comes absorbed evidently into the embryo-sac. For a long time the antipodals have been considered to be merely rudimentary prothallial cells. It is to Westermaier that is due the honour of having proved for the first time their important nutritive function on the basis of anatomical structures and microchemical reactions of various well selected examples.} 1 Westermaier, l.c. 54 T. Ikeda: Osterwalder, in his study of Aconctum Napellus, confirmed Wester- maier’s view on the ground of the particular situation of antipodals, their cytological feature, as well as the structure of the basal portion of the nucellus.!. The opinion of Miss. Goldflus on the antipodals of the Com- positae is also in perfect accordance with Westermaier’s view:? her conclusion is based chiefly on the anatomical structure of the neighbouring tissue of the antipodals. Miss. Balicka-Iwanowska, on the contrary, in studying the embryo-sac of certain Gamopetalae, attributes little im- portance to the nutritive function of antipodals; for she found that ‘‘ the antipodals, when they exist at all, seem to have a transitory function, possess mostly poor contents, and disappear very quickly.”* Campbell,+ in his study of the embryo-sac of Lyszchiton and Sparganium, observed an extraordinary multiplication of antipodals, whereupon he came to the probable conclusion that they must play an important rdle in the nutrition. The divergence of opinions about the function of antipodals is, as already stated in the Introduction, no doubt due to the specific difference of the plants used by the various authors.* For while in some plants they represent the most important nutritive organs, in others they may be mere functionless prothallial cells. Whether they are important in the nutrition is therefore to be decided only upon the basis of detailed studies of each particular case, so that to infer their importance solely on the ground of their extraordinary multiplication, as Campbell does, would not be justified, as Miss. Sargant® states quite rightly in her recent interesting paper. The nutritive function of the antipodals of 7rzcyrtzs hirta will, I think, be concluded on the basis of various facts described till now, though necessarily more or less hypothetically. These facts are :— 1 Osterwalder, l.c. 2 Goldflus, l.c. 3 Balicka-Iwanowska, 1899, p. 68. 4 Campbell, 1899. 5 Guignard says for example as to the Leguminosae: ‘“ Les antipodes disparaissent souvent avant la fécondation, par suite de la résorption du tissue nucellaire sous-jacent ; d’ailleurs leur réle, encore assez problématique, parait terminé peu de temps aprés leur formation; dans d’autres plantes, au contraire, on les voit s’accroitre d’une facon notable, méme aprés la fécondation.”” (1881, p. 200.). 6 Sargant, 1900. Studies in the Physiological Functions of Antipodals, ete. Ur Vr 1.) The phenomena of chromatin-aggregation in the nuclei of the antipodals. 2.) The formation of a bundle of long columnar cells in the portion of the nucellus below the antipodals, the so-called ‘Conducting passage,” which extends to them on one side and tothe special cell-group in the chalaza on the other. On account of the long columnar shape of the cells, as well as their situation in relation to antipodals, etc., they are evidently to be considered as the means of transmission of food material, both carbo- hydrates and protein matter. 3.) The termination of the vascular bundles of the funiculus within the cell-group in the chalaza, indicating the transportation of raw material from the exterior to that place. 4.) The cuticularization of the limiting membrane between the inner integument and the nucellus at an early period, so that food material from the exterior enters the embryo-sac chiefly through the vascular bundles of the funiculus, the cell-group in the chalaza, the conducting passage, and the antipodals.1 It is to be noted that not only do the antipodals elaborate food material for the endosperm-formation, but also for the growth of the egg- apparatus. For in the young embryo-sac, no sooner are the ovum and synergidae differentiated, than the antipodals begin to show their charac- teristic cytological feature, the conducting passage begins to be formed, and at the same time the egg-apparatus is active in increasing its cyto- plasmic contents, so that the material for this growth evidently comes through the antipodals and the conducting passage. When we take into account all these facts, we are led to the con- clusion already stated that the antipodals play a most essential rdle in the nutrition of the embryo-sac, for not only do they absorb raw material and transmit it, but also they elaborate it into a proper form. 1 In various plants investigated till now by many authors for example, in Compositae,.according to Goldflus, the membrane of the embryo-sac is cuticularized, so that all food going to the latter must pass the antipodals, In Zricyrtis hirta, as just stated, the limiting membrane between the nucellus and the inner integument is cuticularized but the cel/-wall of the embryo-sac itself ts not, so that though food materials from the exterior must pass generally through the antipodals, it is not impossible that the nucellar cells forming the lateral side of the embryo-sac, when degenerated, are absorbed directly by the latter. ; 6G T. Ikeda: This conclusion drawn only from morphological facts, is confirmed by microchemical reactions, which are treated of in the next chapter. Microchemical Observations. The microchemical reactions were made on fresh ovules, collected between I-2 p.m. a.) Soluble Carbohydrates. Of soluble carbohydrates the microchemical identification of cane- sugar and glucose in the ovule was tried. According to Molisch,? a- naphtol and thymol were employed for this purpose. After the treatment of free hand sections of ovules, chiefly longitudinal ones, with an alcoholic solution (209%) of a-naphtol under the cover-glass, a few drops of strong sulphuric acid are added. Within at most two minutes, the nucellus, the chalaza and the micropylar region of the inner integument, are coloured deep violet. The vascular bundles in the funiculus as well as in the raphe are coloured at first greenish blue, but become deep violet only after half an hour; this delay of the reaction is probably due to the fact that the penetration of the reagent to the vascular bundles is made possible only after the death of the surrounding tissue. The foregoing experiment is not however sufficient to demonstrate the presence of soluble carbohydrates, because concentrated sulphuric acid may split sugars from glucosides, change starch and cellulose into sugar and thus may give rise indirectly to the same colour reaction. Hence for the purpose of control, living specimens were heated at first with boiling water for a short time under the cover- glass and then treated with a-naphtol and strong sulphuric acid. The reaction in question occurs in the same manner, but markedly later than before, often after half an hour or more, so that it is highly probable that in the living materials of the ovule, there exists at least some kind of soluble carbohydrates. The reaction towards Fehling’s solution was very different from what was expected. When the sections, after the treatment with this reagent, 1 Zimmermann, 1892. 2 Zimmermann, L.c. p. 73. be ee ng ees See ee ee Studies in the Physiological Functions of Antipodals, ete. 57 are carefully and not too strongly heated, until little bubbles begin to be formed, the nucellus, the chalaza, and the inner integument, are coloured slightly violet ; but no red precipitates of copper suboxide are observed, as might be expected. This reaction is no doubt due to the so-called Biuret reaction, caused by the presence of albuminous matter within the cell contents. From the foregoing experiments, as well as from the widely different opinions of various authors, it is impossible to determine what kind of soluble carbohydrates is present in the ovule, but it seems very probable to me that some soluble carbohydrates, such as sugars are there present, which have been transformed from insoluble ones, such as starch; especially the intense violet reaction of the nucellus, the chalaza, and the micropylar region of the inner integument towards Mbolisch’s test, points to the probable occurence of the saccharification-process in these particular por- tions of the ovule. The accounts of this process must be given later. 6.) Starch and Dextrines. In order to ascertain the distribution of starch and dextrines through all the developmental stages of the ovule, the microtome-sections made from materials fixed with absolute alcohol and Keiser’s sublimate and acetic acid mixture, were treated with the chloriodide of zinc, instead of potassium iodide solution, in order to obtain a rapid reaction. Some idea of the distribution of starch and dextrines throughout all stages may be obtained by means of the several diagrams in Pl. VI. In the period of the archespore (Fig. 40) no starch reaction is obtained within the ovule. Starch grains are found in the wall of the ovary, except in its epidermis ; they are probably the products of carbon-assimilation in these places themse!ves, where chlorophyll grains are abundantly present. These starch grains become gradually smaller both in amount and in size towards the ovule, which is entirely free from starch and so is coloured only yellow by chloriodide of zinc. As the development proceeds (Fig. 41), starch grains appear in the epidermis of the ovule, in the funiculus and the raphe, as well as in the inner layer of the outer integument, so that the conclusion is highly 58 T. Ikeda: probable that starch in the ovarian wall penetrates into these parts in some dissolved state, and is here again transformed into starch. Through the period of the development of the ovule, starch penetrates more and more into various parts ef the ovule evidently by the same process, so that the outermost layer of the ovule, the inner layer of the outer integument, the funiculus, the outer layer of the inner integument and the nucellar cap come to possess starch grains, though in the two latter parts the quantity of starch is scanty (Fig. 42). In the earliest stage of the development of the embryo-sac (Fig. 43) the distribution of starch is almost identical with that in the preceding stage, except for the presence of a few grains in the inner layer of the inner integument; but the relative amount of starch is widely different. For, as is shown in Fig. 43, the inner layer of the outer integument and the nucellar cap become richer in starch. At the same time fine grains which are coloured red (somewhat tinged with brown) by the chloriodide of zinc, are found aggregated around the nucleus; they are evidently some kind of dextrine and are derived from starch, which has been absorbed into the embryo-sac in some soluble form. In the following stage (Fig. 44), the distribution of starch in the ovule is nearly the same as in the foregoing. In the embryo-sac, the dextrine granules are visible in both the egg-apparatus and the antipodal cells. Starch in the inner integument disappears and minute dextrine granules appear in the micropylar region of the inner integument. After the union of the polar nnclei (Fig. 45) the distribution of starch is almost the same as that in the foregoing stage, but the number of dextrine grains deposited in the tissue of the inner integument surrounding the micropyle, gradually increases. These granules are found in the parietal cytoplasm of the embryo-sac as well as in the ovum, but they are never observed in synergidae. Later, when the embryo-sac is ready for fertilization (Fig. 46), the relative distribution of starch grains in various parts of the ovule still remains unaltered, but their amount in the funiculus and the raphe has been largely augmented, while at the same time their great decrease in the outermost layer of the ovule and their certain decrease in the inner layer ot the outer integument, is to be noticed. The amount of dextrine granules Studies in the Physiological Functions of Antipodals, ete. 59 in the micropylar region of the inner integument attains its climax in this stage ; besides the amount of them within the egg-cytoplasm has gradually increased. As to the dextrine granules aggregated in the micropylar region of the inner integument, it is very probable that they serve for the nutrition of the pollen-tube, which makes its way into the micropyle along the ovarian wall. Various reasons speak for this probability : they attain their maximum amount just before fertilization (Fig. 46) and disappear soon after it; also the fact of the non-cuticularization of the micropylar region of the inner iutegument (cf. the foregoing chapter) is in favour of this hypothesis. When fertilization is over and the endosperm nucleus begins to divide (Fig. 47) the distribution of starch grains becomes gradually modified. In general, their amount decreases, especially in the outermost layer of the ovule and the inner layer of the outer integument. The dextrine granules deposited {in the egg-cytoplasm increase in amount concurrently with development of the embryo; they are probably used for the purpose of ege-nutrition and kept in the cytoplasm as reserve materials. When the formation of the endosperm proceeds, the decrease of the reserve starch grains in various parts of the ovule is immense. Except the small quantitiy in the outer integument and the pretty large amount in the funiculus and the raphe, no starch grains are found. The amount of reserve dextrine in the embryo increases gradually. The fertilized ovum remains undivided, until an immense quantity of the endosperm is produced. At this stage of the endosperm-formation (Fig. 48), the antipodal cells already show signs of disintegration and remain attached to the endosperm as flattened discoidal pieces. The supply of nutriment for the purpose of endosperm-formation is furnished by the conducting passage, already in the process of degeneration. Even in this stage, no starch grains are visible in the endosperm. When ripening is near (Fig. 49), the relative distribution of starch becomes distinctly altered. No starch or at most only a trace of it is found in various parts of the ovule, except in the inner integument and the endosperm tissue where it is abundantly present. On account of the growth of the endosperm, the funicular portion and consequently its vascular 60 T. Ikeda: bundles and cortical parenchyma now become so extremely compressed as to obliterate their lumen. The passage of nutriments from outside would not therefore take place through such elements; the only possible way is probably the epidermis. The only noticeable fact, which has not taken place in all the fore- going stages, is the immense aggregation of starch in the inner integument. This phenomenon, which is seen only in the latest period of the endosperm- formation, has possibly some connection with the subsequent development of the endosperm after the supply of food from outside has been stopped. The aggregration of starch grains in the endosperm begins at first in the micropylar end of the embryo-sac and then proceeds toward the chalazal end. From what has been described up to this point, it will be seen that starch has never been met with in the chalaza, the antipodals, or the whole nucellus (including the conducting passage) except the nucellar cap. As already stated, starch formed in cells of the ovarian wall is transported into the ovules, where it is again transformed into the original form and deposited in various tissues of the ovule. This reserve starch afterwards undergoes again the chemical change into soluble form and is transported into the embryo-sac. The place where the diastatic enzyme, which inter- venes during this transformation, is formed, is the cell-group in the chalaza, which, as above stated, is characterized by the smallness and scanty cytoplasmic contents of its constituents; and water necessary for this hydrolysis may be supplied by the vascular bundle and especially by the spiral tracheides, which terminate amid the chalaza in somewhat knot- like enlargements. For the same reason, starch in the ovarian wall may change into soluble form by the action of diastase, which probably is secreted by a special cell-group near the placenta, characterized also by the smallness and rich cytoplasmic contents of its constituents ;1 water necessary for its hydrolysis may be supplied also by spiral tracheides, which run through the centre of this group of cells. The chalaza and the conducting passage, which are always free from 1 Billings (1901) has recently applied to this cell-group the name of “nutritive tissue” (,, Nahrgewebe”). Studies in the Physiological Functions of Antipodals, ete. OI starch, show the ‘intense violet reaction in the living state, so that they possess soluble carbohydrate instead of starch. This result is in contradiction to the statement of Westermaier, who always found starch in the conducting passage, so that in our case the name ‘‘starch route”’ (,,Stirkestrasse”) given by him to this tissue must be changed into ‘‘ sugar route.” Fertilization. The so-called ‘‘ double fertilization” takes place in Trceyrtzs hirta. No generative nuclei with coiled or vermiform shapes as discovered by many authors were observed in my preparations, but as the male nuclei in their free state in the embryo-sac were not found, it is not yet decided whether they are not vermiform from the beginning, as is the case with Exdimyon investigated by Guignard.! Besides the accurate observa- tion of both sperm nuclei in the pollen-tube was not possible on account of the too deep staining of the contents of the latter. The two generative nuclei discharged from the pollen-tube take their course to unite with the egg-nucleus and the primitive endosperm nucleus respectively. Though direct observation is wanting, it is almost undoubt- ed that the considerable changes in the structure, size, and shape of these generative nuclei must have taken place during this transition through the embryo-sac. The egg-nucleus, ready for fertilization, is always flattened or lenticular in its optical section; it has a single nucleolus in its centre, and is scanty in chromatin. The ovum is poorer in cytoplasm than the synergidae, but is always richly loaded with dextrine granules (Fig. 50) ; even after the fusion with the male nucleus, no change in the physical consistency of cytoplasm and nucleus in the ovum takes place. The nucleus of the synergidae, which is greatly reduced in size just before fertilization, lies in the dense granular cytoplasm. The union of both germ nuclei is at first a mere apposition; the apposed nuclei become flattened against each other and compressed toa certain degree, so that the amount of chromatin seems to show a 1 Guiguard, 1899. 62 T. Ikeda: relative increment. The size and shape of the male and female nuclei in the state of apposition are almost identical. No structural difference was observed between them, except the absence of the nucleolus in the former (Fig. 24, ¢.z. and g.z. rz). The septum of the apposed nuclei then gradually fades away and the fusion of both nuclei is accomplished. The resulting nucleus with a single nucleolus as before becomes more and more spherical (Fig. 25). The egg-cytoplasm possesses the alveolar structure as before fertilization and then the ovum grows in size. It remains entirely or almost without growth long after the fertilization, even till after the formation of many nuclei from the endosperm nucleus (Fig. 33). During this development, the nucleolus of the egg-nucleus becomes divided into two or three pieces and prepares for the subsequent division. The synergidae, one of which was observed to be still alive at the time of fertilization, after it, undergo, sooner or later the process of destruction. The second generative nucleus discharged into the embryo-sac makes its course towards the primitive endosperm nucleus, and during this time an enormous change in its size and shape, as well as in its structure, seems to take place. The second generative nucleus is much larger than the other, being sometimes equal to that of the primitive endosperm nucleus (Fig. 24, . and g.z. 2.). Its shape is also similar to that of the latter, being sometimes ellipsoidal or cone-shaped. The paternal and maternal chromatin elements of the resulting nucleus are distinguishable long after the fusion. The absence of the true nucleolus in the generative nucleus is here also to be noted. Vacuoles are sometimes present. The primitive as well as the definite endosperm nucleus is extraordinarily large and the single nucleolus is of huge size. The fusion nucleus is always irregular in contour ; and towards the side of fusion there seems to be a denser aggregation of chromatin. Formation of the Endosperm. The most remarkable facts brought out in the study of the endosperm- formation are the manner of its formation and the behaviour of the endo- sperm nuclei. Studies in the Physiological Functions of Antipodals, ete. 63 After a certain period of rest the definitive endosperm nucleus begins to divide according to the ordinary mode of mitosis. During these changes, the giant nucleolus becomes gradually reduced in size (Fig. 27 @ and 6) and finally disappears. The outline of the two resulting daughter nuclei is very irregular, always taking more or less long rod-like shapes, and we find here at the beginning several, usually spherical nucleoli (Fig. 28) Then they divide both at the same time (Fig. 29) and thus give rise to four daughter nuclei (Fig. 30). In the latter nuclei as well as in those derived by later divisions, we find in the resting stage the nucleoli of various sizes. They are at first spherical but later they develop pseudopodia-like processes (Fig. 30) and begin to break up into several pieces (Fig. 31, 33, 34), so that there are often seen some nucleolar fragments scattered within the nuclear cavity. As chromatin begins to increase the nucleoli begin to draw back their pseudopodia-like processes. Even in the advanced stage of nuclear division nucleolar fragments are found scattered near the nuclear spindle, (Fig. 36 a). The whole behaviour of the nucleolus above described then corresponds no doubt to what was observed by Zimmermann! during the nuclear division in various plants and led him to the erroneous conclusion ,,Omnis nucleolus e nucleolo.”? For example, our figure 36 a corresponds exactly to his figure 32 (cell from the stem-apex of Pszlotum triquetrum), and also our figure 32 somewhat resembles his figure 27 (cell from the root-apex of Vzctza faba) or 38 (spore-mother-cell of Eguisetum palustre). During these periods of development, the nuclei themselves become elongated and assume various remarkable forms, resembling those con- cerned in amitotic division (Fig. 34). Their mitotic division was often met with, where the arrangement of chromosomes in the equatorial plane was pretty irregular (Fig. 36 a and 4). The peculiar forms assumed by the endosperm nuclei, as above described, might perhaps give rise to the erroneous conclusion that they were concerned in amitosis, but as real mitosis was observed repeatedly, it is highly probable that this phenomenon is to be considered as that of 1 Zimmermann, 1883. 2 Zimmermann, 1896. p. 64. 64 T. Ikeda: surface extension for the purpose of metabolic interchanges between the nucleus and the cytoplasm, Phenomena which might be included in the same category have been observed in both animal and vegetable cells. Of the former, we may cite the well-known discovery of Korschelt in the water-beetle Dytzscus: the egg contains at a certain period the dense granular nutritive masses, which are believed to have come from outside ; the germinal vesicle becomes amoeboid, sending out long pseudopodia, which are always directed towards the principal mass of granular sub- stances.1. Of the vegetable cells, Kohl? observed in the living nuclei of marginal cells in the leaves of Elodea canadensis, as well as in those of the hair-cells in leaves of T7radescantia virginica, that by the action of an asparagine solution these nuclei are incited to make amoeboid movements, and he attributes this phenomenon to an energetic interchange between the nuclei and the cytoplasm. The endosperm nuclei, thus formed by the mitosis are uniformly distributed throughout the granular, compact cytoplasm, and after a long time when the embryo has become nearly ripe the membranes are formed between these nuclei, and thus the formation of the endosperm is completed. Even when seeds are near ripening, these nuclei are characterized by their curious shape (Fig. 37); they are scanty in chromatin and furnished with several round nucleoli, but finally they take the usual shape and undergo the ordinary mode of karyokinesis. Each endosperm cell posses- ses scanty cytoplasm, but is filled with an immense amount of starch grains (Fig. 39a, 6). The formation cf cell-membranes between these endosperm nuclei begins at the micropylar region of the endosperm and proceeds towards the chalazal portion. This always happens after the full agegre- gation of cells with starch. The embryo remains very small and shows no differentiation, even when the seeds are almost ripe (Fig. 38). From what has been described before, we see that the endosperm formation of 77zcyrtis differs widely from the ordinary course. For itis a 1 See Wilson, 1900, p. 349-350. : 2 Kohl, 1897. ‘Studies in the Physiological Functions of Antipodals, ete. CN Ur well-known fact that usually during this process the cytoplasm of the embryo-sac forms at first a thin parietal layer, where successive nuclear divisions take place, and then afterwards the hollow inner space is gradual- ly filled up. But here the embryo-sac is fromthe very beginning filled up with dense cytoplasm and the nuclei formed by successive divisions are scattered in it uniformly. Such a mode of development seems, according to Lloyd,1 to take place also in Vazlantia hispida belonging to the Rubiaceae, Summary. I. The archespore arises as usual from a subepidermal cell and develops directly into an embryo-sac-mother-cell. Then two successive divisions occur and thus four cells are formed, of which the lowest one develops into the embryo-sac, while the three above become obliterated. 2. Soon after the maturation of the embryo-sac, the union of two polar nuclei takes place. 3. The antipodals are prolonged downwards towards the funnel- shaped haustorial part of the embryo-sac at its chalazal end. The nucleus of the antipodals is at first scanty in chromatin, but soon it begins to show the phenomena of chromatin-aggregation. This is due to metabolic activity, so that when their activity approaches its end, the chromatin- masses begin gradually to dissolve away. 4. Ofthe nucellar cells, which are placed at the basal portion of the embryo-sac, those of the axial row soon take a long columnar shape and form the so-called ‘‘conducting passage.” According to the microchemi- cal test, the latter is always free from starch and seems to contain soluble carbohydrates. The conducting passage continues toa special cell-group in the chalaza, in which the vascular bundles of the funiculus terminate. These vascular bundles pass through another cell-group near the placenta and unite with the main trunk of the vascular bundles of the ovary. When we take these anatomical structures as well the microchemical reactions into account, we can imagine the mode of transmission of starch. It is trans- formed into soluble carbohydrates by diastase secreted probably by these 2 Lloyd, 1899. 66 T. Ikeda: cell-groups near the placenta and the chalaza, then the soluble carbo- hydrates pass through the funicular vascular bundles and the conducting passage respectively and are absorbed into the antipodals, which either elaborate them there into their proper form or transmit them to the proper place. 5. The nucellar cells surrounding the conducting passage degenerate probably on account of enzymes secreted by the antipodals. These products of degeneration may be absorbed directly by the antipodals or indirectly through the conducting passage and are used for the nutrition of the embryo-sac. The immense number of granular masses met with in the conducting passage is derived probably from these degeneration- products. . 6. All these facts—the cytological features of the antipodals, the anatomical structure of the neighbouring tissue, especially the formation of the conducting passage, as well as the results of microchemical tests— all justify the conclusion that the antipodals in Zyzcyrtis hirta are the centre of the absorption of raw materials, their elaboration into the proper form and the means of the transmission of food to the proper place. 7. During development, dextrine granules tare deposited in the antipodals, the ovum, the parietal cytoplasm of the embryo-sac, and in the micropylar region of the inner integument. They are evidently the reserve material. As to the dextrine granules in the micropylar region, there are various reasons for the hypothesis that they serve for the nutri- tion of the pollen-tube, which passes through this region. ’ 8. The so-called ‘double fertilization” takes place. Whether the generative nuclei are vermiform or not, is not yet decided. 9. During the endosperm-formation, the nuclei take various curious shapes. This is probably for the purpose of surface extension, due to the metabolic activity between cytoplasm and nuclei. 10. The mode of endosperm-formation differs greatly from the ordi- nary one in this that the embryo-sac is from the beginning filled with the compact cytoplasm and successive nuclear divisions occur within this cytoplasm. Studies in the Physiological Functions of Antipodals, ete. 67 / This work was carried on in the Botanical Laboratory of the College of Agriculture of the Imperial University of Tokio, under the guidance of Professor Ikeno, to whom I have therefore in the first place to express my sincere gratitude. I am also indebted to Prof. Ishikawa for his valuable counsel throughout the progress of the work. Se 68 T. Ikeda: LIST OF PAPERS QUOTED. Balicka-Iwanowska, G.: Contribution a I’étude du sac embryonnaire chez certain Gamopétales. Flora 86, 1899. Billings, F. H.: Beitrige zur Kenntniss der Samenentwickelung. Flora 88, I9OI. Campbell, D. H. : Notes on the Structure of the Embryo-sac in Spargantum and Lystchiton. Bot. Gaz., 27, 1899. Goldfius, M.: Sur la structure et les fonctions de I’ assise épithéliale ct des antipodes chez les Composées. Journ, de Bot. 12—13, 1898—18g9. Guignard, L.: Recherches sur le sac embryonnaire des Phanéroga angiospermes, Ann. disc. nat. Bot, Vi, 13, 1neee Sur Porigine du sac embryonnaire et le réle des antipodes. Bull. de la Soc. bot. de France, 28, 188r. Les découvertes récentes sur la fécondation chez les végétaux angiospermes. Volume jubilaire de la Soc. de Biologie. 1899. Quoted in Sargant, 1¢0o. Hacker, V.: Praxis und Theorie der Zellen-und Befruchtungslehre. Jena, 1899. Huie, Lily, H.: Changes in the cell-organs of Drosera rotundifolia, produced by feeding with egg albumen. Quarterly Journ. of Micros Se. 30; 1807, Further study of cytological changes produced in Daosera. Part Ly Lb: Az) rage: duel, H.0.: Beitrige zur Kenntniss der Tetradentheilung. Jahrb. f. wiss. Bot. 35, 1900. Kohl, F.G.: Zur Physiologie des Zellkerns. Bot. Centralb., 72, 1897. Lloyd, F. E.: The comparative embryology of the Rubiaceae. Memoirs of the Torrey Botanical Club, 8, 1890. Magnus, W.: Studien iiber die endotrophen Mycorrhiza von MWeottia Nidus avis L. Jahrb. f. wiss. Bot. 35, 1900. Osterwalder, A.: Beitrige zur Embryologie von Aconitum Napellus L. Flora. 85, 1898. — Studies in the Physiological Functions of Antipodals, ete. 69 Rosenberg, 0.: Physiologisch-cytologische Untersuchungen tiber Drosera rotundifolia L. Upsala, 1899. Sargant, E.: Recent work on the results of fertilization in Angiosperms. Ann. of Bot. 14, 1900. Schniewind-Thies, J.: Beitrige zur Kenntniss der Septalnectarien. Jena, 1897. Review by Mobius in Bot. Centralb., 69, p. 216—218. Die Reduktion der Chromosomenzahl und die ihr folgenden Kerntheilungen in den Embryosackmutterzellen der Angiospermen. Jena, Igolr. Westermaier, M.: Zur Embryologie der Phanerogamen, insbesondere uber die sogenannten Antipoden. Nova Acta Acad. Leop.-Carol. 57, I. 1890. ——: Zur Physiologie und Morphologie der Angiospermen-Samenknos- Peleaeoecitr.. Zz. wiss, Bot, I,.2. 1896. Wilson, E. B.: The cell in development and inheritance. 2nd. ed. New York, 1900. Zimmermann, A. B.: Die Botanische Mikrotechnik. Tiibingen, 1892. : Beitrage zur Morphologie und Physiologie der Pflanzenzelle, IT, 1. Tubingen, 1893. Die Morphologie und Physiologie des pflanzlichen Zellkernes. Jena, 1896. 70 Fig, Fig. Fig. > se Fig se Fig. Fig. Fig. 5 Fig. Fig. Fig. Fig, Fig. T. Ikeda: EXPLANATION OF FIGURES. ——___0 +e —____ PLATE I. 1—10. Various stages of the first nuclear division of the embryo-sac-mother-cell, Fig. 9 and 10, Zeiss, homogeneous immersion ;45 and ocular 3, all others the same obj. and oc. 4. 11. Second nuclear division of the embryo-sac-mother-cell. 3X5. 12. Beginning of the embryo-sac with three degenerating sister-cells, 3X Js. 13. Embryo-sac with two nuclei. 2X 4. 14. Embryo-sac with two polar nuclei. 3 7s. 15. Two volar nuclei concerned in copulation. 2X5. 16. Mature embryo-sac. 0, ovum; syz., synergidae ; avt., antipodals, Nucleolus of primary endosperm nucleus extremely large! 4X +5. 17. Cells of the conducting passage with protein granules, avz., antipodals. 3X4. 18. One antipodal short after fertilization, Chromatin-aggregation in the nucleus! 4X75 to. Two antipodals, Later stage than that of Fig. 1& Chromatin-aggregation less remark- able, Fibrillar structure in cytoplasm! 4-5. 20. One antipodal later than in Fig. 19, Fibres in cytoplasm now becoming very distinctive. Leitz. 4X. PLATE: I. 21. Cross-section of an ovary with two ovules longitudinally cut. c#., chalaza; /., faniculus ; 0, W., ovarial wall; #7. cell-group in the placenta. 2XA. g. 22, a. The same in cross-section, showing the cell-group in the placenta. 2xD.b. The cell- group in the placenta much magnified. 2, . 23. Cross-section of an ovale. ax. c¢., axial cell-group ; 7. f., parietal layer of the nucellus ; z, , inner integument ; 0. 2., outer integument ; ef., epidermis. Leitz 4X4. g. 24. Double fertilization. ¢. ., nucleus of the ovum; g. 7. Z, first generative nucleus ; g. 7. 2 second ; ., primary endosperm nucleus. 3X75. . 25. Second generative nucleus and primary endosperm nucleus in fusion, ‘The fertilization of the ovum already accomplished. 3s. . 26. Secondary endosperm nucleus in spireme stage. 2 x 45. . 27. a,b, The same in anaphase. Two consecutive sections. 3 xy. 28. ‘Two daughter-nuclei derived from the first endosperm division. 3 x 7s. . 29. ‘Two above daughter nuclei dividing. /. ¢., pollen-tube ; 0, ovum, 2x 4s. . 30. Endosperm formation, Nucleoli with pseudopodia-like processes. 4 x 75. 31. Thesame. Advanced stage. Nucleoli fragmenting. . 32. Thesame. Advanced stage. Nucleus having peculiar shape. 4x js. . 33. Thesame. Nucleoli fragmenting. 0, fertilized ovum. PLATE III. . 34. Endosperm-formation, Nuclei of various remarkable shapes. 4 x #s- Tig. 35. ‘The same. Nuclei in spireme stage. 4x ly. Studies in the Physiological Functions of Antipodals, ete. Fig. 36. Thesame. Nuclear division. a. side-view ; b. polar view. 3 x 7s. Fig. 37. Thesame. Cell-walls already formed. Nuclei of remarkable forms. 4 x 5. Fig. 38. Embryo in almost ripe seed ; no differentiation! 4,4. Fig. 39. Nuclei in later stage of endosperm formation, a, Resting nuclei. b. Upper cell with the nucleus in dispireme stage ; in the lower one the longitudinal division of chromosomes figured. Cells filled with starch grains. 4x 4. PLATE IV. Figs. 40—49, schematic representations of various stages of development of the ovule. Abbreviations : o.w., Ovarial wall; ¢,.7., external integument ; a., archespore ; 7.7. #z., micropylar region of the inner integument ; Z. ., polar nucleus ; /. ¢. 7.. primary endosperm nucleus ; 7s. ¢., nucellar eap; z. z., inner integument ; 7., nucellus ; 0. ¢., epidermis of the ovule; cz., cuticularized membrane: ch, chalaza; f., funiculus; ov., ovum; syz., synergidae; e. 7., endosperm nucleus; ant, des., disorganized antipodals; des. e., disorganized sister cells of the embryo-sac; 7%.. raphe ; Z¢., pollen-tube. Fig. 4o.. Archespore formation. Tig. 41—42. Stages of the embryo-sac-mother-cell. Fig. 43. Embryo-sac formatiou. Fig. 44. Polar nuclei not yet united. Fig. 45. Polar nuclei already united. Dextrine granules in the ovum and around the synergidae. Fig. 46. Fully matured embryo-sac. Fig. 47. Short after fertilization, Fig. 48. Endosperm formation, Fig. 49. Later stage of endosperm formation. Fig. 50. Ovum with dextrine granules, CONTENTS. PAGE, Introductory... css sec. See lace Sumy Gamal papi One| ce oleate Materials and. Methods... .<. 0 sae wwe: jee xus) Geen 9 sae geen Formation of the Embryo-sac.°... “<2. sss sea) @yues seu ee Maturation of the Embryo-sac... 2.10 - seam egtes oe) nae ee Antipodals; Intesuments, Nucellus, Chalaza-ete) | ssaeocaie sean eee a. Antipodal Cells’... 2. 00 ee 6. Integuments, Funiculus and Raphe. -% | 2 =:-. «=e é. /Nucellus.and Chalaza..... ¢s.. ces, ese) ee | ) Microchemical Observations... :.. «2: ven” vec eeu a. Soluble Carbohydrates. ... iste toc) Sen SSR 6. Starch and Dextrines.. ... -... sc. sa. ee Fertilization. 220 ese i. wes wee “ese ben cen et Formation of the Endosperm. ... see) cnc Sen Sue SUMMIT 00. cies es ade) eae y cus, oem Ses sole eer List of Papers Quoted: «2.0 vd ta “inn ae Explanation of Figures... ... © ss. 01. tos) ORe Seen! BULL. AGRIC. COLE. VOL. V: Auctor del PLATE Wf eT atl. O.---------- to* ae) el AY BULL. AGRIC. COLL. VOL. V. AG Auctor del. GATE IVS = PLATE: ¥ BULL. AGRIC. COLL. VOL. V. —- PLATE Kel BULL. AGRIC. COLL. VOL. V. as ‘ : ohhh it at tn Oe, KIS ik Desi at 3S nS : Ln) & a & Ss ~~ a) (scanty). Do. T7ve. Dextri 3 > Ss larized membranes. «Cytoplasmic con Lew me Cut tents. - = ' { re = ‘ ‘ . 4 Contributions to the Study of Silk-Worms. I. ON THE EMBROYOLOGY OF THE SILK-WORM. BY K. Toyama. (With Plates VI[—XL.) Since the publication of Tichomiroff’s beautiful work on the ‘‘ Dévelop- pement du ver a soie du murier dans Voeuf,” about ten years ago, no important facts have been added, so far, as I am aware, by any renewed researches, except a short description by Graber ('90). The observations which are described in the following pages, were carried on in the Zoological Institution of our College during last year, and though they may not bring to light anything new, yet they are here published with the hope that they may be found to be of some value. Before going further, I wish to express my heartiest thanks to Prof. Ishikawa for affording me much assistance and advice in the present investigation, and to Prof. Sasaki for kindly placing at my disposal various publications out of his own library. IWViethods. Eggs deposited on a karton are killed by the solution (1—2) of chromic acid or by the Flemming’s weak triacid solution, both heated to 80—go°C. The latter reagent is fitted for young stages, while the former is used only for advanced ones. A great number of eggs hardened by hot corrosive sublimate, picro-sulphuric or picro-acetic acids, proved to be useless. The eggs treated by the chromic or Flemming’s acids for two or more hours, are thoroughly washed with water and transferred for harden- ing to 70% alcohol in which they remain for several weeks or even months. 7p K. Toyama: By this process the content of the egg which adheres closely to the shell is shrunk a little and a narrow space is left between the surface of the egg and the shell, so that we can easily remove the shell with dissecting needles. It may be noted here that the eggs treated by the chromic acid solution are preserved in the most satisfactory manner, both the nuclei in resting and in karyokinetie states being beautifully brought out and the yolk being made capable of being cut without difficulty, so that a complete series of sections can be obtained. Staining is done on the slide with alum-carmine, glycerine or alum haematoxylin, and gives beautiful results in all stages as will be seen from the figures, which are all drawn from these preparations. A. My own observations. I. The formation of mesoderm and entoderm. In our country, univoltine silk-worms generally deposit their eggs in June or July. Five or six hours after deposition, the first division of the segmentation nucleus will be observed in the middle of the anterior portion of the egg. Some of the increased nuclei migrate towards the surface where they form the blastoderm at the end of about one day, while the others which remain in the interior of the yolk become vitellophags. If we cut an egg at the end of the second day after its deposition, the ventral plate will be seen completely detached from the other portion of the blastoderm. In this stage, the lower layer (Unteresblatt) is not yet completely formed and the inward growth of cells will be seen to take place from the primitive furrow at the anterior portion of the ventral plate as has been already described and figured by Tichomiroff.* The closure of the blastopore takes place very slowly, as will be seen in an embryo represented in Fig. 1, Pl. VII., where it appears as a wide furrow tapering at both ends (Fig. I, bl. a). This embryo was killed at the end * Tichomiroff 1. c. Fig. 16. Contributions to the Study of Silk-Worms. 75 of August or September. A circular depression will be observed at the anterior end of the furrow (Fig. 1, 0). The internal developmental processes of this stage are shown in Figs. 2, 2°, 2°, 3,4, 5 and 5°. Fig. 2 is a transverse section passing through the anterior end of the blastopore. The ectoderm invaginates here in the form ofa deep sac. A similar invagination at the primary head segment was also observed ina younger stage than this by Tichomiroff ('82) who represents it in his Figure 14, but has failed to give its fate. Further development brings the lips of the invaginated pocket close together in the median line (Fig. 2") thus forming a closed sac, with an irregularly shaped lumen which is compressed dorso-ventrally. This corresponds to the ‘“‘ primordial Spalte”’ of Heider. Fig. 2” shows the median longitudi- nal section of this stage, in which the lumen is distinctly visible. The cells composing the wall of the sac gradually become more and more irregular and wandering out into its lumen completely fill it up, and form a loose cell-mass. These changes are noticeable when we compare Fig. 2 with Figs, 2” and 3. On closer examination, we see that the mass is composed of large round cells of soft and succulent appearance and with aclear outline (Fig. 3). Some of these cells will frequently be seen to detach from the mass, as is shown in Figs. 7, 7* ms”. The invagination of the ectoderm just described is only observed in the primary head segment where the circular depression above mentioned is situated (Figs. 1 and 6, o, 0’), while in the mandibular or maxillary region the inner layer is formed by an inward growth of ectodermal cells from the base of the shallow blastoporous furrow, as will be seen in Fig. 4. In a more posterior portion of the embryo where the blastoporous opening is wide (Fig. 1, a.), the inner layer is formed by a lateral overgrowth of the ectoderm as is seen in Fig. 5. And in the anal segment we observe a wide and shallow ectodermal furrow from the bottom of which the migra- tion of cells takes place to form the inner layer (Fig. 5', pf). This may be regarded asa modified form of the deep invagination that is seen at the anterior end of the blastopore. Not infrequently, in this stage, we meet with cells which detach themselves from the lateral portions of the ectoderm and enter the yolk- 76 K. Toyama: mass (Fig. 5°, par). Tichomiroff (‘82) has also observed these immigrat- ing cells and has represented them in his Figs. 14 and 15, m'. He considers them to be the secondary entoderm and comes to the following conclusion :— “Pendant les premiers stades de développement, cet entoderme secondaire, au moment méme de sa formation, se convertit immediatement en mésoderme.” According to my observations, however, they correspond to the “ Paracyten” of Heymons ('95) and like them gradually disintegrate into smal! granules having no share in the formation of mesoderm. Migratory cells of similar nature were also observed in many insects by Graber. Although the ectoderm does not as yet show any sign of segments in this stage (Fig. 1), certain alterations are already found in the inrer layer. The most important of these is its metameric arrangement. This process begins at the middle portion of the germ-streak and proceeds both, forwards and backwards, as has already been observed by Tichomiroff. In the present stage, there are 17 or more segments faintly marked off from one another, and among these the first which is derived from the deep invagination of the blastopore, as above mentioned, is the. largest. At the end of November, when the embryo attaines the stage given in Fig. 6, the blastopore is nearly closed, being represented only by a faint line, except at the posterior half (Fig. 6 a), where it remains some- what open. The cells on both sides of the central cell-mass at the anterior end of the blastopore (Fig. 7.ms'), are now separated from it and form a mass of small-cells (ms) closely attached to the ectoderm. These are the mesoderm cells and are now easily to be distinguished from the cells of the central mass by their smaller size and by their nuclei being deeply stained. The cells of the central mass are, on the contrary, larger and, of a spherical form, and their cytoplasm is very much vacuolated staining faintly with haematoxylin or carmine (Fig. 7 a), thus making them easily distinguishable from the others even under a low power. “Cells are seen to detach from the central mass in this as in the preceeding stage, and to migrate into the yolk-mass (Figs. 7 & 7a ms*). They differ, Contributions to the Study of Silk-Worms. NI N however, entirely from the vitellophags or “ Paracyten” in the structure of the nucleus and also in form and size. The central cell-mass thus far considered may be compared with the endoderm-anlage of Hydrophilus as is described by Heider, or with that of Doryphora as given by Wheeler, but it is of quite a different nature as will be shown later on, and for this reason we will call it the oral cell-mass. An interesting question on this point is: Whether the oral cell-mass will remain as a definite tissue? To this we shall come later on. After the closure of the blastopore, there remains a round ectodermal depression in the middle of the primary head segment where the oral cell-mass is situated (Figs. 6, 70'). This is a structure of a transitory nature, disappearing in a more advanced stage where it is only represented by a shallow median furrow (Figs. 12 and 130’). In the maxillary or thoracic region, the inner layer has already separated off from the ectoderm asa distinct layer (Fig. 8 ms), while the median ectodermal furrow still exists (Fig. 8, pf) which closely resembles the neural furrow. As we go towards the posterior, however, the boundary of the ectoderm and the inner layer gradually obliterates (Fig. 9) until finally the inner layer becomes exposed to the surface (Fig. 10). In the anal segment, the wide furrow above mentioned (Fig. 5%, pf) becomes narrow and the two distinct layers, the ectoderm and the inner layer, are again formed (Fig. 11). Moreover, in this stage, the inner layer is heaped up inthe median portion, its lateral arrangement becoming visible only when the segmentation of the ectoderm begins to appear. Soon afterwards, the narrow groove extending from the oral depres- sion to the anal segment fades away with the closure of the posterior opening of the blastopore,. but no trunk segment is as yet to be seen (Fig. 12). It isin this stage that the embryo passes the winter, namely from December to the end of January, or the first part of February. Among the hundreds of embryos we studied, we did not find one that had passed beyond this stage. Other varieties of the silk-worm, such as the bivoltine, 78 K. Toyama: multivoltine etc. also pass the winter in this stage, so that we may call it a resting stage. Figs. 13—16 represent transverse sections through an embryo ot this stage, i.e. the resting stage. . The first section (Fis. 03) @jaasses through the primary head segment; the oral cell-mass (Fig. 13, ms’) elongates considerably into the yolk, and at its distal portion some immigrating cells (ms*) are to be seen. Besides these, we first meet with other cells migrating in the yolk-mass such for example as degenerat- ing cells (d.c) which will be considered later ina separate section. The next sections (Figs, 14 and 15) pass through the anterior portion of the thoracic region, the former representing the segmental portion, while the latter the intersegmental. The segmental arrangement of the mesoderm is now distinctly visible. These segments are 18 in number, the first (the oral cell-mass) and the last (the anal segment) being the largest (compare Figs. 13 and 16, with Figs. 14 and 15). This reminds us of Wheeler’s Fig. 72, Pl. XX., which represents a longitudinal section of an embryo of Doryphora, of which he says: ‘‘ These two masses of cells are the independent sources of the entoderm, which grows backwards as two strings from the anterior mass and forwards as two strings from the posterior mass.’ These cell-masses in Bombyx mori, however, are not the entoderm-anlage as we shall see further on. The warm days of spring awake the embryo from its winter sleep. It now increases greatly in length, and with this the procephalic lobe extends more laterally. A number of outer segments also make their appearance developing in number posteriorly. With these changes of the external parts, the internal portions also change. The median mesoder- mal cell-mass flattens and spreads out below the entire ectoderm and finally becomes divided into two lateral streaks (mesodermal streaks) by the withdrawal of its cells from the median line (Fig. 26, Pl. VIII). The cells constituting the lateral streak are clearly of two layers, the upper consisting of cylindrical cells arranged regularly, while the lower layer consists of flattened cells arranged irregularly. These evidently cor- respond to the ‘‘paradermalen” and the ‘ paralecithalen Schicht” of Heider. a . a Contributions to the Study of Silk-Worms. 79 One of the most interesting changes in this stage is the disintegration of the oral cell-mass to form migratory cells. This is seen in Fig. 17, which is a longitudinal section through the primary head segment. Here the cells are clearly seen migrating from the periphery of the oral cell- mass (Fig. 17, ms’). Tichomiroff who first observed this cell-mass in this stage (see his Fig. 17) considers it to be the mesoderm-anlage saying that ““nous voyons que le premier des dix-huit segments anterieurs du meso- derme differe par sa forme de tous les autres. Ce segment tiré son origine de la partie la plus profonde du sillon primitiv. Les cellules different egalement du reste du mésoderme : elles sont un peu plus grosses que les cellules mesodermiques ordinaires et leur plasma est plus clair.” Moreover, he homologizes it with the structure which Hatschek (’77) for the first time found in Bombyx and considered to be the entoderm-anlage (a structure which in reality is not the entoderm-anlage but the subcesopha- geal body) and concludes with the following words: ‘‘ Nous allons voir quen réalité il n’en est rien; cet epithelium (Middarmepithelium) a une origine differente.” The further changes concerning the cell-mass are not, however, given. Fig. 18 is a surface view of a more advanced embryo in which the neural furrow has made its first appearance as a faint median line. In the primary head segment, we again meet with a wide depression. In this stage, we observe not infrequently the bifurcation of the neural furrow at the anal segment, resembling closely the bifurcation of the blastopore at the caudal end of the Xiphidium embryo as is observed by Wheeler (’93), or of the Lina embryo observed by Graber (’90). Fig. 73 which represents a cross section at this portion of the embryo shows the two ectodermal furrows atthe bottom of which cell-proliferation is to be observed, but the true nature of these furrows is as yet quite obscure and requires further study. Let us now consider the oral cell-mass. Its fate will be best under- stood when we examine Figs. 19—26, which are a series of transverse sections passing through the primary head segment of an embryo in the Same stage as that represented by Fig. 18. In the anterior portion (Fig. 19), the mesoderm flattens out on both sides of the median line 80 K. Toyama: forming two lateral masses. These gradually approach each other as we go posteriorly until they join} together in the median line and with the oral cell-mass (Figs. 20—21). Figs. 22-26 are consecutive sections of the oral cell-mass in which the disintegration of its cells into migratory cells will be clearly seen. The immigration of cells begins at the anterior periphery of the oral cell-mass (compare Figs. 17, 22, 23, and 3), and proceeds gradually to its distal end which elongates consider- ably into the yolk (Fig. 27) and forms the two arms of an inverted Y (in Fig. 28 only the left arm of the Y is represented). The cells usually detach from the distal end of the arms one at a time but in some cases also in groups (Fig. 22 ms’). Some important changes are to be observed in the embryo shortly after the preceding stage (Fig. 20): cephalic and thoracic appendages now become distinctly formed as lateral outgrowths of their respective segments, The antennae (at) originate as lobular outgrowths from the posterior edges of the procephalic lobes. The stomodial depression (st.) now distinctly appears and from its anterior edge the labrum (lb) will be seen as two separated processes. The three thoracic segments are very slightly or not at all broader than the two maxillary segments. The appendages of these six segments are also nearly alike in shape, size, and position except that those on the mandibular segment are larger than the others (Fig. 30 md). The mandibular appendages also differ from the others in this that they are directed horizontally, while the other appendages are placed latero-posteriorly. The space between the primary head segment and the mandibular segment is occupied by ganglion cells (Fig. 30 vk) and represents the ‘‘ Vorkiefersegment ” of German authors. Proctodium now appears also in the form of a faint and shallow depression at the posterior end of the anal segment (Fig. 70 a). Returning now again to the consideration of the oral cell-mass, we find that the disintegration of its cells becomes more vigorous with the ingrowth of the stomodium until at last the entire cell-mass completely disappears. Thisis clearly to be seenin Figs. 29, 31 and 32. Fig. 29 is a median longitudinal section of the primary head segment where Contributions to the Study of Silk-Worms. Sr a shallow stomodial depression (st) makes its first appearance just’ in front of the oral cell-mass. In Fig. 31 the stomodial depression is more marked and with this the curvature of the ectoderm is increased, which evidently accerelates the detachment of the oral cell-mass from the ectoderm. And when the stomodial tube becomes more elongated, and its terminal portion becomes broader asin Fig. 32, not even a remnant of the cell-mass is to be seen. From what has been above described, we may safely conclude that the tnvaginated cell-mass at the antertor end of the blastopore, although zt greatly resembles the entoderm-anlage of other authors such as Heider, Wheeler etc., ts certainly not to be regarded as such in the present case. Then the questions naturally arise: (1) Whence arises the entoderm? ; (2) What is the oral cell-mass and the cells migrating fromit? The first question will be considered of in the next paragraph ; while the second will be discussed later on under the heading, ‘‘ The vitellophags and cellular elements found in the yolk-mass.”’ The formation of the mid-gut (Mitteldarmanlage.) As before said the stomodium makes its first appearance as a shallow ectodermal depression at the anterior portion of the oral cell-mass which represents the anterior end of the blastopore. As the development of the embryo advances the depression gradually deepens and proceeds backwards along the ventral wall of the embryo, as is shown in Figs, 38 and 39. At the two lateral corners of its free ventral end the elonga- tion of the stomodial ectoderm takes place (Figs. 37, 38 a). These ectoderm-elongations correspond to the ‘‘vordere Epithellamelle”’ of Voelatzkow and Heymons, who first observed them in Coleoptera, Ortho- ptera etc., and they give rise to the formation of the epithelium of the mid-gut. The epithelial cells of the mid-gut originate, as has just been said, from the ectoderm-elongations at the posterior end of the stomodium, and have nothing to do with the cells of the blastopore, which latter do not, as has been said before, now exist asa definite tissue. Nor do the mesoderm cells of the ‘‘ Vorkiefersegment” have any relation to 82 K. Toyama: the formation of the mid-gut epithelium. These are beautifully visible in Figs. 33—40, which represent a series of sagittal sections of the embryo ‘given in Fig. 30. In the embryo given in Fig. 30, the stomodial tube takes an oblique ‘course along the ventral wall as already referred to, and the ventral apex (2) of the tube is more elongated than its dorsal corner (¢) (Figs. 37, 38, 39). It will also be seen that the anterior portion of the ectoderm of the ‘‘Vorkiefersegment” is taken into the formation of the ventral wall of the stomodium and helps the curving of the ectoderm at this place. The mesoderm attached to this portion gradually detaches itself from the ectoderm and proceeds posteriorly along the ventral wall of the stomodium, forming a structure which was first discovered by Hatschek ('77) in Bombyx, and erroneously considered to be the entoderm-anlage. Wheeler ('‘93), who also found this same structure in Xiphidium, has given it the name of the subcesophageal body. Tichomiroff (‘82) also observed this body in a more advanced silk-worm embryo, calling it by the name of the ‘‘corps adipeux du seconde ordre,” and derived it from the central mass of yolk-cells. When fully formed, its cells are large and it stains more faintly by carmine or haematoxylin than any other structures found in the body of the embryo. The cytoplasm is very granular and has a distinctly yellow tint even in the unstained sections, With the elongation of the stomodial tube the suboesophageal body proceeds backwards until it becomes situated in the ventral side of the fore-gut within the methothorax. Figs. 41—51 represent transverse sections through the primary head segment of an embryo taken out from the same deposit as the one just described, but a little more advanced than the former. Fig. 41 passes through the anterior portion of the primary head segment; Fig. 42 six sections behind it, and in front of the stomodium. Here the ectodermal depression is more developed and its median portion is elevated into a ridge (a). The depression becomes deeper as we go backward (Eig, 43), and its lateral lips come close together in the median line until a tube is formed (Figs. 44—47) which is compressed dorso-ventrally. Between the ectoderm and the stomodial tube are seen some free cells (Figs. 46, Contributions to the Study of Silk-Worms. 82 47, 6c) which represent blood cells. It may be here remarked that the compressed lateral edge of the stomodium (Figs. 45—47, a) elongates somewhat laterally as a distinct tissue. This portion corresponds to the elongation of the ventral wall of the stomodium already refered to (Figs. 37—39), and becomes more and more developed concurrently with the development of the embryo, and the epithelium of the mid-gut is formed by the proliferation of this tissue as will be seen in the following pages. In the vicinity of the distal end of the stomodium we observe many free cells in the yolk (Figs. 49—51). We can not accurately determine the origin of these cells. They may arise from the oral cell-mass or some of them may come from the mesoderm. We believe, however, that they arise from both the oral cell-mass and the mesoderm. What- ever their origin may be, we are certain that ‘they take no part in the formation of the entoderm. We specially directed our attention to this Poimt, but we were not able to meet with even a single case in which the formation of the entoderm by these wandering cells could ‘have occurred. Like all other insects that have a stage during which the body is greatly elongated (Fig. 18), the silk-worm passes into a series of stages during which the germ-band is gradually shortened (Fig. 52). The shortening is accompanied by a broadening of all the segments, a growth of the appendages, and very important internal changes ; the cephalic and ‘thoracic appendages have meanwhile assumed a more definite character. The first and second maxillae and thoracic appendages have each become three jointed. The abdominal appendages now also make their appearance on the first ten segments with the exception of the anal, as Kowalevsky ('71) and Tichomiroff ('82) long ago observed in Lepidopterous insects. Graber ('88), however, doubts the observations of Kowalevsky and erroneously states that Tichomiroff did not discover them. But, as is stated above, we are not only able to say that abdominal appendages do really exist in silk-worms, but we can also confirm the statement made by Packard on this point, that ‘‘these structures appear in the embryos of certain Lepidoptera and Hymenoptera, though they are much less distinct and more evanescent than in the lower orders of insects.” 84 K. Toyama: It is interesting to note here, moreover, that the stigma appears in: each segment from the first thoracic to the eleventh. In the meso-and’ metathoracic segments we are able to observe faint depressions of the ectoderm, which may represent the rudiments of stigma. Of these the one on the mesothorax together with the stigmata on the last two- abdominal segments disappears entirely, while the remaining ten pairs. of stigmata persist in the larval stage. The rudimentary stigmata on the metathorax does not disappear, but remains as a small opening without external chitinous ring and internal closing apparatus, such as the closing lever, closing band, or closing bow. ‘Tichomiroff ('92) has given a quite correct description of these metathoracic stigmata in saying that ‘‘le stigmate du metathorax ne disparait pas, il demeure sous la forme d’um. stigmate rudimentaire avec son faiscean correspondant trachéal, méme chez la larve adulti; ce stigmate rudimentaire se trouve fortement avancé- vers le mesothorax.” We are now able to add that these rudimentary stigmata do not only persist in the larval, but also in the imaginal stage, in which they are clearly recognizable. Their internal structures are, however, different from those on the prothorax or abdomen, wherea stigmata is provided witha closing bow, a closing band, and a closing lever, as is described by Krancher (‘81) in Smerinthus, whereas the metathoracic stigmata have only the closing bow with its well developed muscles. Recently Boas (‘99) has observed the presence of stigmata in each thoracic segment in the larva of Cossus ligniperda, which, however ends: blindly. We can not detect any trace of closed stigmata in silk-worms. Fig. 54 isa median longitudinal section of the primary head segment of the embryo above described. The stomodium has now become longer than in the preceding stage and its distal end widens out. The wall of the stomodium consists of thick epithelium, except at the bottom where it becomes quite thin (gl.), constituting the ‘‘Grenzlamelle”’ of Heymons. The ‘“‘vordere Epithellamelle” has developed more than before and close to its ventral side the subcesophageal body (sb.) appears as a distinct mass of cells. A cross section through the antennal region of an embryo in the same stage, is shown in Fig. 53. The stomodium, as already described, ‘Epithellamelle. Contributions to the Study of Silk-Worms. Cc Ur ‘is found to be a compressed tube ; its lateral edges (ent) are thicker than its dorsal and ventral walls, and project into the yolk as distinct structures. “This is the anlage of the entoderm already referred to. Attached to the ventral wall of the stomodium we again meet with the subcesophageal body. But the development of the entoderm-anlage will become clear ‘when we come to examine the serial sections mentioned below. Fig. 55 isa longitudinal section through the primary head segment -of a slightly more advanced stage, showing the elongation of the ‘‘ vordere i] Series of transverse sections of the head segments of this stage are given in Figs. 56—63, showing more clearly the relations between the lateral projections of the stomodial wall and the epithelium -of the mid-gut. The first section (Fig. 56) represents the section ‘through the bottom of the stomodial invagination whose ventral wall ‘presents here three thick folds, on each side of which will be seen lateral projections of cell-mass (ent) very well developed while the dorsal ‘wall, which represents the cross section of the “ Grenzlamelle,” is very thin. The stomodium becomes more aud more compressed as we proceed posteriorly (Fig. 57), until in the section represented by Fig. 58 the lumen of the stomodium has completely disappeared and only a flat -cell-mass resting on the subcesophageal body is to be seen. In this flat cell-mass we again observe the thick lateral portions (ent) which have been already referred to. Inthe section passing though the anterior ‘portion of the mandibular segment (Fig. 59), we observe only thes lateral cell-masses (ent) and a portion of the subcesophageal body (sb) attached to the ventral sides, while the median portion has entirely -disappeared. In the next section here represented (Fig. 60), which passes through the posterior portion of the same segment, the subcesophageal body vis no longer observed, and the lateral cell-mass or the entoderm-anlage only are left attached directly to the lateral mesoderm (Fig. 69). In these mesoderm masses we can not make any distinction between the splanchnic -and the somatic portions, all the lateral masses consisting of irregularly shaped cells (Fig. 60 ms). In the maxillary segments (Figs. 61, 62) the dorsal end of the lateral mesoderm forms a curved compact tissue con- sisting of one layer of cells, somewhat resembling the head of the figure 86 K. Toyama: 8. The curved-inner end of this portion of the mesoderm (Figs. 61, 62, 63, sp. ms) represents the splanchnic layer, and the outer portion (Figs. 61, 62 and 63, sm. ms) the somatic layer, but a closed coelomic cavity is not formed. On the dorsal andthe inner portions of the splanchnic part of the curved end of the mesoderm will be seen the entoderm-anlage orthe prolongations of the lateral cell-masses of the stomodium, which. in the anterior part consists of irregularly shaped cell-masses (Figs. 61, ent). The cells of these masses are large and contain vacuoles in the cytoplasm. Inthe posterior part, however, they consist of high columnar cells arranged in a single layer and firmly attached to the inner dorsal portion of the splanchnic mesoderm (Fig. 62, ent). The entoderm-anlage become smaller as we go posteriorly until in the second thoracic segment (Fig. 63) they are represented by only a few cells, attached to the splanchnic mesoderm. In the yolk-mass in the vicinity of the distal end of the entoderm- anlage we meet with, not infrequently, small cells containing small but deeply coloured granules. These are to be seen in Fig. 64, which is. a cross section of the sixth abdominal segment. On closer examination, they are found to be nothing else than degenerating cells, the function of which is probably to give nutrition to the growing entoderm. These cells together with blood cells are also found in other places in the yolk, but mostly in the neighborhood of the entoderm (Figs. 62, 63, and 64). In his Fig. 45 (which corresponds to the stage shown in my Fig, 52) Tichomiroff distinguishes ‘‘ trois trabécules de cellules vitellines ” ; namely a middle and two laterals, and is of the opinion that the neurilemma and the fat bodies are derived from the former, while the epithelium of the mid-gut comes from the latter. On this point he says ; ‘ils donnent naissance & l’epithelium de l’intestine moyen, épithelium que se constitue des cellules de l’entoderme secondaire, émanant de ces trabecules des cellules.” We have also frequently observed such cells corresponding in all particulars to those givenin his Fig. 54 Ef. (see our Figs. 59, 64, and gt dc), but we have not been able to find any such cells in any stage of transition into the entoderm, On the contrary, we frequently Contributions to the Study of Silk-Worms. 87 meet with these cells disintegrating into small granules and showing the phenomena of degeneration. From all that has been thus far said, we may conclude that they are degenerating cells and have nothing to do with the formation of the epithelium of the mid-gut. Fig. 65 isa frontal section of the posterior portion of the stomodium of a more advanced embryo. We observe init the karyokinetic division of the nuclei of the epithelium of the mid-gut, by which the cells of this region are multiplied thus causing its closure. From the foregoing accounts, we may sefely conclude that the anterior entoderm-anlage ts formed by the proliferation of the epithelial cells of the stomodium, and ts consequently ectodermal in its nature. Netther the mesodermal cells formed by the cells of the blastoporous tnvagination, nor the yolk cells, have anything todo with the formation of this structure. The proctodium and the Malpighian vessels. The first appearance of the proctodium is always later than that of the stomodium, as has already been observed by many other authors. Even when the stomodial depression becomes as long as is represented by Fig. 30, or 31, the proctodium is seen only asa faint depression at the posterior end of the neural furrow (Fig. 70%). Its median longitudinal section is represented in Fig, 71. Different from the stomodial depression, it is directed somewhat towards the dorsal side and forms a round tube, the blind end of which consists of thick epithelium (Fig. 72), where an active proliferation of cells will be observed (ent). These cells give origin to the posterior entoderm. The lateral wall of the blind end of the proctodium will now be seen to give rise to short evaginations (Fig. 72, uv), which ultimately become the Malpighian vessels. As regards the formation of these structures, Tichomiroff states that ‘‘ ces derniers (Malpighian vessels) ne se présentent en effet que comme de simples excroissances tubulaires de l’intestin posterieur. Comme on sait une larve adulte posside six vaisseaux malpighians, qui debouchent dans lYintestin posterieur par deux conduits communs. Ce sont ces conduits ——— | - | | 88 K. Toyama: qui apparaissent’ des le commencement comme de simples excroissances de l’intestin posterieur, se partagent ensuite chacum en trois tubes comme les trones trachéaux principaux émanant des stigmates, se divisent en > Hatschek ('77) also observed three tubules on rameaux secondaires.’ each side of the proctodium or hind-gut of Bombyx, and says that ‘‘ die drei malpighi’schen Driisen jeder Seite miinden durch ein gemeinschaft- liches Anfangsstiick in das blinde Ende des Hinterdarmes.” According to our observation, these arise as three separate pairs of hollow outgrowths from the beginning as will be seen in Fig. 76, which is taken from an embryo a little more advanced than that represented in Fig. 72. As the entoderm and the Malpighian vessels appear just at the same time, the position of the entoderm-anlage is disturbed and the study of it by cross sections becomes very difficult. Fig. 74 is a sagittal section of the proctodium ofa more advanced stage, which corresponds to that shown in Fig. 52. Here we observe that the proliferation of the cell-mass from the ventral wall of the proctodium assumes the form of a short lamella and is directed forwards forming a ‘‘hintere Epithellamelle” (eplh). When the revolution of the embryo is about to set in, this becomes more elongated and a dorsal process is formed (Fig. 75). From the entoderm-anlage, two lateral stripes grow out and assume the form of a U, the arms of which are directed forward and become attached to the splanchnic mesoblast in just the same way as the anterior entoderm-anlage. This is shown in Fig. 54, which shows the foremost portion of the posterior entoderm. In this section, we observe some genital cells (g) placed in the somatic mesoderm, from which they are differentiated. This is what has been shown by Wheeler in Xiphidium, and by Heymons in Phyllodromia. Although the clusters of germ cells are normally seen to occur in the third and the sixth abdominal segments, we often observe them in all other abdominal segments with the exception of the anal ; and in one case, we observed them even in the mesothorasic segment. Weare thus in position to say that the genital cells originally arise in each body segment. Tichomiroff also found genital cells in a far more advanced stage than this, and first maintained the opinion that they were derived from the entodermal cells, ‘‘mais apres m’étre familiarisé plus tard * S he Contributions to the Study of Silk-Worms. 89 avec le rdle important que remplit l’entoderme secondaire dans la formation de different organes, jincline a admettre que la partie essenticlle des organes sexuels se constitue aussi & ses depens.’’ But they originate from the mesoderm as the former facts show. Summary: 1. The posterior etoderm-anlage 1s derived first from the epithelial wall of the proctodium tn the same way as the anterior entoderm-anlage ts derived from that of the stomodium. 2. Malpighian vessels arise as three separate pairs of outgrowths from the blind end of the stomodium. 3. Genttal cells differentiate from the cells of the Somatic mesoblast. II. The vitellophags and other cellular elements found in the yolk. a. Witellophags. Just as in the eggs of many other insects thus far studied, we here also find in the yolk various cellular elements among which the most conspicuous are the vitellophags, or cells left in the yolk atthe time when the other cleavage cells are traveling towards the surface of the egg to form the blastoderm. In certain cases, however, all the segmentation- nuclei come to the surface of the egg, and in these cases the vitello- phagous cells are formed by the migration of the cells of the blastoderm, as was observed by Patten ('84) in Phryganids, by Uzel ('97) in Campodia, and in many other cases. In the silk-worm, the vitellophags are formed from the segmentation- nuclei which are left in the yolk. When the segmentation of the yolk- mass is completed, we find a single vitellophag in the centre of the yolk. This multiplies by direct division (Fig. 68) when the embryo attains the stage given in Fig. 1, so that in more advanced stages we see many nuclei in the centre of each of the yolk-segments (Fig. 9, yd). The cytological nature of these vitellophags is as follows :—The body ofthe cell is large as compared with other cellular elements found in the yolk. It has a large round nucleus with fine granular chromatin Re go K. Toyama: scattered init. There is no nucleolus to be seen. The cytoplasm is stained faintly in the younger stages of embryo, while in later stages it produces numerous pseudopodial processes (Fig. 69 vit) and stains well either with haematoxylin or carmine; the fine granular chromosome becomes thicker. Thus they are readily distinguishable from other cellular elements in the yolk and are easily seen (Figs. 2, 7, 9, 22, 24, 25, 22.33 etc:). The vitellophags are very often seen collected in the vicinity of the entoderm-anlage (Fig. 61) or other organs, but they have never been observ- ed to transform into other tissues. We frequently observe, however, that their nuclei become irregular in shape and about to dissolve (Fig. 67 vit), and although we have traced their metamorphosis up to the stage when the embryo hatches, yet we have failed to find any direct evidence of their forming other organs, and we can definitely say that they take no part in the formation of the mid-gut-anlage or any other organs, but degenerate in situ and finally undergo dissolution. b. Migratory cells from the ectoderm. In younger stages we frequently observe cells which are about to detach themselves from the ectoderm (Figs. 2%, 5', par). These are small ellipsoidal cells with a point like nucleus (Fig. 19, par). These cells probably correspond to the ‘‘ Paracyten” of Heymons, who described and figured them in his beautiful work ‘‘On De1maptera and Orthoptera”’ (‘95). Tichomiroff ('82, '92) has also observed these cells but considers that they become the mesoderm. It is, however, certain that these as Heymons (‘95) rightly says, have no direct share in the formation of any embryonal tissue, but finally dissolve away. ec. The cells migrating from the oral cell-mass. As already stated, the oral cell-mass disintegrates and forms migra- tory cells. In the early stages of the embryo, we observe them in the vicinity of the oral cell-mass (Figs. 7, 13, and 17 ms’). In later stages Contributions to the Study of Silk-Worms. QI the oral cell-mass sends out two branches from its free end, as already stated (Figs. 27, 28 ms’). These elongate into both sides of the dorsal portion of the embryo where some liquid content is to be seen (Fig. 27 c). Fig. 28 is a magnified drawing of a portion of the left side of the embryo as shown in Fig, 27, and shows the separation and metamorphosis of the cell-mass into free cells. These cells are round, ovoid, or spindle-shaped, having a large nucleus which contains dense chromatin-granules and one or two nucleolei (Fig. 28 ms’). The cytoplasm contains numerous vacuoles in its periphery, while it is dense near the nucleus, staining deeply (Fig. 28 ms’, Figs. 67, 69, ms’). Sometimes we see that these cells become irregular in outline and stain faintly by haematoxylin or carmine, and finally dissolve as shown in Fig. 67. As to the function of these free cells, Schwartze ('99) is of the opinion that these give origin to the blood corpuscles. Although they resemble blood cells in their general appearance, yet we have no direct proof that they are such, and we are inclined to think that they are nutritive cells which have the function of liquefying the yolk and conveying it to other portions of the egg, and that they finally dissolve. This appears the more probable when we learn that some of these free cells pass out from the body of the embryo and wander about the extra-embryonal yolk- mass ; and also that with the increase of these free cells, degenerating cells containing smail granules increase suddenly in number in the yolk near the oral cell-mass. d. The blood cells. As already stated, we observe a different structure in the anterior end of the mesoderm which afterward becomes the subcesophageal body (Figs. 38, 39, 4g a). The mesoderm at this stage of development consists of irregularly shaped cells having homogenous cytoplasm which stains uniformly, except at the anterior end where the cells are seen to detach from the rest (Fig. 40 a). These cells are vacuolated and stain very faintly. In amore advanced stage in which the subcesophageal body is well 92 K. Toyama: formed, we observe at the same place some free cells of a circular form, containing vacuoles in the cytoplasm and staining faintly except at the nucleus (Figs. 47, 54, 55, b.c). Karyokinetic divisions (Fig. 54 a) are sometimes to be seen among them. These cells are blood corpuscles, but as the number of them produced from this portion of the mesoderm is small, it is probable that they are formed from. other portions of the mesoderm in a similar way; this latter point however we are not able to state definitely at present. In Figs. 20, 21, 22 b.c’ we see a number of cells separating off from the mesoderm at other portions of the body, which we had considered to be the blood corpuscles. More careful examinations, however, convinced us that they were in reality not such, but rather degenerating cells which will be described under the next heading. e. Degenerating cells. Ina younger stage of the embryo, there are only vitellophags and a few ectodermal migratory cells in the yolk-mass. In the spring, when the ventral plate begins to elongate and the disintegration of the oral cell-mass becomes vigorous wandering cells in the yolk-mass gradually increase in number, especially in the vicinity of the primary head and anal segments, as was observed by Wheeler in Doryphora (Figs. 17, 22—25). With the increase of wandering cells, we also observe various cells containing small granules which are stained well by haematoxylin or anilin colors. They may be observed everywhere in the yolk-mass near the germinal streak, but like the other wandering cells they are especially abundant in the vicinity of the primary head and anal segments, as shown in Figs. 43—45, and 74, 76, d.c. At the time when the stomodium, the proctodium etc, begin to be formed, these cells suddenly multiply in such numbers, that they are easily distinguishable even under a low magnifying power. In the beginning, however, they closely resemble other migra- tory cells, and contain stainable round granules in the cytoplasm Contributions to the Study of Silk-Worms. 93 (Figs. 13 a, €6 d.c). These granules gradually increase in number and with this the nucleus dissolves, till at last the celis are entirely filled up with the granules (Fig. 69 d.c). The size of the cells is not uniform, some being large and others small. Finally the cells disintegrate and the granules alone are left freely suspended in the yolk-mass. Where do these degenerating cells come from? They may either arise 1. from the migratory cells of the ectoderm; 2. from those of the mesoderm ; or 3. from the free cells of the oral cell-mass. 1. That they arise out of the migratory cells of the ectoderm is to be seen when we compare Figs. 2,7 5! with Figs. 13, 19, 22 par. In Figs. 2,75% we see cells (par) about to detach themselves from the ectoderm and these cells have an appearance similar to certain small cells in the yolk (Figs. 19, 22 par) which are certainly to be recognized as the degenerating cells referred to above, both by the presence of granules in their cytoplasm as well as by their general form and size. 2. their origin from the mesodermal cells can be clearly seenin Fig. 96 where cells of an exactly similar appearance are met with. 3. Lastly, that they arise out of the’ oral cell-mass is very plainly to be seen from Fig. 69 where these degenerating cells are seen in the neighborhood of the free cells of the oral cell-mass and among them cells of the intermediate condition are often to be distinguished. Moreover, in the spring when the disintegration of the oral cell-mass most vigorously takes place, a sudden increase of the degenerating cells is to be observed as already described. The function of these cells is difficult to state. But it will not be far fetched if we consider that they give nutrition to the growing portions of the embryo, such as the proctodium, stomodium etc. From the foregoing statement, we are able to say that there are four sorts of migrating cells in the yolk. The first are the vitellophags derived Jrom the remainder of the segmentation nuclet. The second are cells separat- ed from the ectoderm and undergoing degeneration. The third are cells migrating from the mesoderm. Some of these become blood corpuscles while others degenerate like the second. Lastly, the fourth are the cells 94 . K. Toyama: which are produced by the disintegration of the oral cell-mass. These also undergo degeneration. III. The endoskeleton of the head, with reference to the salivery gland and a new gland. The first trace of the endoskeletons of the head appears in the embryo shown in Fig. 52. In this stage, we observe many ectodermal invagina- tions in the lateral part of the mandibular and the maxillary segments. Figs, 77—82 are serial sections through the head segments showing these invaginations. The first section (Fig. 77) passes through the anterior portion of the mandibular segment in a somewhat oblique direction. In the right side of the section, we see the antenna and the mandible, from the outer base of which an invagination of the ectoderm (Fig. 77 tent’) takes place, while in the left side where the knife passed through the middle of the mandible we observe the tubular structure of the above invagination. Posteriorly we observe a new tubular invagination on the inner side of the basal portion of the mandible at its posterior portion (Fig. 78 fl. md). As we proceed more posteriorly we again meet with such an invagina- tion on the lateral sides of the first maxillary segment (Fig. 79 tent”). A similar invagination also takes place inthe next segment at the outer base of the second maxillae (Fig. 80n). Another pair of invagina- tions is also to be seen at the inner base of the second maxillae (Fig. 80 slg). The latter are the origin of the silk glands. Figs. 81—82are the consecutive series of sections next to the section shown in Fig. 80. Here we see posterior portions of the two invaginations at the second maxillary segment. The fate of these four pairs of tubular invaginations excepting the silk gland, will be seen in Figs. 83—90, which represent a series of sections through the head of a more advanced embryo, in the stage shortly before the revolution. In this stage the head segments are about to . Contributions to the Study of Silk-worms. 95 coalesce with one another and the limit of each segment is no more visible (woodcut Fig. 1), Fig. I. tent’, first and second tentorium; pie ex. md, attachment of extensor mandi- an bulae; fl. md, attachment of flexor ex. md md. mandibulae. Ib, labrum; an, anten- oe mx?, nae; mx’™, first and second maxillae ; - = mx?, sl, g, silk gland; n, new gland or hypo- th, 12 a MS stigmatic gland; th. I", first-third th. 12 e # thoracic legs; H-shaped dotted line, tentorium anlage within head. sl. g. Figs. 83 and 84 represent two consecutive sections through the anterior portion of the head. The invagination between the labrum and the mandible (tent’), exactly corresponding to the invagination figured in a younger embryo (Fig. 77 tent’) can be clearly observed. The invaginat- ed tube becomes flattened and proceeds posteriorly along both sides of the stomodium and unites with the invaginated tube at the first maxillary segment (Fig. 89 tent’, right side and wood cut, dotted line) which proceeds forwards along both sides of the stomodium. So we see on both sides of the stomodium two parallel tubes opening at both ends, one at the base of the mandible andthe other at the base of the first maxilla. These tubes afterwards unite with each other by producing transverse processes from both tubes and together form a H-shaped tube (woodcut Pie 1, dotted line). These tubes are the anlage of the tentorium of the head. The walls of the head, consequently, are supported or braced within by beams resembling an H which correspond exactly to the tentorium described by Tichomiroff in the head of a silk-worm (Tichomiroff Fig. 35). Returning again to the mandibular region, we see an invagination at the outer base of the segment (Fig. 85 ex. md ; woodcut Fig. I. mx. md). This becomes the seat of the muscle extensor mandibulae. It is short and small, and is the last invagination which takes place in the head. At 96 ; K. Toyama: the posterior inner end of the mandible we again meet with a large invagination (Fig. 86 fl. md, and woodcut Fig. I. fl. md), It is a flattened tube curved at its anterior portion, so that in a cross section it presents a form like a crescent (Figs. 87, 88, 89 fl. md), while at its posterior portion the tube is circular and slender (Fig. 90 s.g). | This anterior portion chitinizes afterwards and becomes the seat of the flexor mandibulae, while the posterior portion gives origin to the salivary glands. These relations will be seen more clearly if we compare the accompanying sagittal section (Fig. 92). Here it will be seen that the invaginated tube sends off a large branch which is directed anteriorly, and the mesodermal cells are largely accumulated around it to form the muscles. Posterior to this branch it proceeds as a round tube (s.g) which becomes the salivary gland. Thus the seat of the muscles, flexor mandibulae, and the salivary gland arise from the same invagination at the posterior base of the mandible. The invagination at the lateral part of the second maxillary segment above described (Fig. 80) will now be considered. In the shortening of the head-segments to form the head, the appendages on the second maxillary segment become fused together and form a triangular process. This proceeds more forwards and enters between the appendages on the first maxillary segment, as is shown in the woodcut Fig. I. mx*. The two openings of the silk glands come close together and become a single opening, ,while the lateral invagination forms a long cell-mass suspended from the ectoderm into the body-cavity. In consequence of the shortening of the second maxillary segment, the greater portion of it is now situated in the pro- thorax (see woodcut Fig.1). In the transverse section we observe these cell-masses on both sides of the subcesophageal body in the prothorax (Fig. 90 n), while in the sagittal section the first portion of them appears as an invaginated tube (Fig.92n). Fig. 94 represents a frontal section through the head andthe thoracic segments in an embryo a little more advanced than the above. It will be seen that the cell-body and also the nuclei of these cells are larger than those of the surrounding tissue and stain somewhat more deeply. This cell-mass persists as a definite body in the larval stage. It is flat and trilobed, resembling fat-tissues in Contributions to the Study of Silk-Worms 97 appearance, and is situated on the ventral side of the first stigma to which itis firmly attached, while its front end is attached to the hind edge of the head. Fig. 95 represents the cell-mass of a larva at the end of the third stage. Inthe full grown larva, it also exists but it becomes more elongate and produces more branches, as is shown in Fig. 96 which is taken from a larva of the fifth stage. Its lengthis now about 16mm. and its breadth 0.09 mm. This body resembles the fat tissue in general appearance, but it can very easily be distinguished from it by its cells, which are large and contain a dendritic nucleus (Fig. 97), while the cells of the fat tissue are small, their nuclei circularand the cytoplasm mostly with fat granules. The function of this body is quite obscure, the structure of the nucleus, however, assures us that it is a glandular organ representing perhaps a sort of dermal gland such as the cenocytes or dorsal glands which are not in the prothorax, the other segments of the body are provided with one or two such. But as we have thus far been unable to find any gland of this description mentioned in the literature on insect- anatomy, we will call it ‘‘ the hypostigmatic gland.” If we now cive a short summary of the above statements it will be as follows: I. Ln the mandibular segment, three pairs of tuvaginations take place; the most anterior (between the labrum und the mandible) becomes the first tentorium, the second pair gives rise to the seat of the extensor mandtbulae, while the last becomes the flexor manhibilae and salivary gland. 2. Inthe first maxillary segments, there ts a pair of invaginations which become the second tentoriun. 3. Inthe second maxillary segment, we again meet with two pairs of tnvaginations, the inner of which forms the silk gland, while the lateral ones grow intoa gland which ts situated on the tnner side of the first stigma in the larva, and which we will call the hypostigmatic gland. 98 K. Toyama: B. General considerations. Let us now consider the results obtained by other investigators as compared with those given above. The mesoderm. As regards the development of Lepidoptera, Kowalevsky ('71) was the first naturalist who clearly described the formation of the mesoderm in Sphinx populi. He noticed that the mesoderm was not formed by the typical groove-shaped invagination of the ectoderm, but by a pair of lateral overgrowths. A comparison of our Figs. 5 and io with his Figs. 5 and 6, Taf. XII., will show that the mode of the formation of the mesoderm is the same in both cases. Next to him comes Bobretzky ('78) whose observations principally concern the formation of the blastoderm, but also give some description of the formation of the germinal layers. According to this author the formation of the mesoderm takes place in Lepidoptera later than in other insects, namely after the formation of the embryonal envelopes, the amnion and serous membranes and ‘“‘tritt in Form einer seichten, langlichen Rinne auf, deren Bodenzellen, sich vermehrend, sich vom Keimstreifen abbilden.” From this we may say that in this species of Lepidoptera studied by Bobretzky, the mesoderm is formed by an inward growth of cells from the bottom of the blastoporous groove. Tichomiroff (82, '91) in his interesting article above referred to, ‘‘ Development du ver a soie du murier dans l’oeuf” describes and figures the formation of the germ layers. Concerning the mesoderm formation, he maintains the opinion that ‘‘ elles procédent, avant tout, de l’ectoderme et 2) de l’entoderme.” A true invagination tube at the anterior end of the blastopore and an inward growth of cells from the median primitive furrows takes place, which forms the mesoderm (see his Figs. 14 and 15). Bruce’s ('87) observation on Thyridopteryx also shows the inward growth of cells from the primitive furrow to form the mesoderm, and Contributions to the Study of Silk-Worms. exe) his Fig. VII., Pl. I., corresponds exactly with our Fig. 2 on this point. He says: ‘‘ The inner layer is not strictly invaginated, for itis cut off from the rest of the embryo before the opposite sides of the median groove have met,” and ‘‘ the median groove deepens, beginning to push dorsally the median portion of the embryo. In subsequent stages, the groove deepens, and the pushed-in-portion of the embryo becomes folded off and forms the inner layer.” Graber ('90) also observed the inward growth of the cells of the median ectoderm to form the mesoderm in Pieris and in other Lepidoptera, while at the intersegmental region it invaginates as a deep furrow. But the cell-mass which he calls the ‘‘ Ptychoblast” in his Fig. 129 Pt, seems to me to be the oral cell-mass found in other Lepidoptera as above mentioned, and not the inner layer. Lastly Schwartze ('99) has recently published a valuable paper on this point in Lepidoptera; the following is quoted from his description of mesoderm formation :— ‘“‘Die Bildung des mesoderms ist bei Lepidopteren nicht in ein bestimmtes Schema gebunden, sondern erfolgt bald durch Einsenkung eines Rohres, bald durch Zellwucherung von Boden einer Rinne aus, bald durch seitliche Uberschiebung ; es kommen sogar in den verschiedenen K6rperregionen desselben Embryo verschiedene Form der Mesoderm- bildungen vor.” The results obtained by my investigation on silk-worms as above given quite confirm the opinion of the last named author. The deep invagination in the anterior end of the blastopore closely corresponds to a similar groove onthe cephalic lobe described by Bruce and figured in his Fig. VIII. Pl. 1., while the inward growth of cells from the bottom of the me- dian furrow inthe mandibular or in the maxillary segment greatly resembles that described in the observations made by Bobretzky, Tichomiroff, Bruce, Graber et al. Lastly the formation of the mesoderm by the lateral overgrowth of the ectoderm at the median portion in the abdominal segments confirms again the observation made by Kowalevsky. | } 100 K. Toyama: The entoderm. Inthe interpretation of the insect-gastrula the entoderm has always played an important réle. The origin of the mesoderm has long been known in a general way, but the true origin of the lining membrane of the mid-gut has not yet been completely ascertained; some authors maintain the opinion that this originates from the yolk cells, others think that it comes from the ento-mesoderm, while still others derive it from the ectoderm. My first intention was to give a comparative description of the ento- derm formation in the different orders of insects, but as this has been treated ina masterly manner by Heymons ('95) and Schiwartze (‘98) we shall confine our remarks mainly to the Lepidoptera. In the Lepidoptera, Tichomiroff was the first who minutely described the entoderm formation. In his first paper ('79), he considers that it arises from the ento-mesoderm, but in later papers (‘81, ‘91) he maintains the opinion that it is derived from the yolk-cells, that is, the secondary yolk- cells derived from vitellophags by division, as referred to above. Bruce ('87) was certainly at fault when he considered the formation of the entoderm in Thrydopteryx as being formed from the inner layers, the formation being described by himin the following words: ‘‘a portion of the inner layer on each side of the embryo becomes separated from the other parts of the inner layer. These portions of the inner layer which may be called entoderm grow together and unite first on what is the ventral surface of the alimentary tract.” Ritter ('90) is also similarly mistaken in his study of Chironomus. Graber (‘90) made many valuable observations on the bipolar origin of the entoderm in Bombyx, Pieris, Gastropacha etc. Although he has not given any direct proof on this point, yet from the fact that in many Lepidopterous insects, the division of the yolk-cells does not occur, or occurs only after the formation of the entoderm, he comes to the con- clusion that the entoderm is not formed out of the yolk-cells. Comparing other insects, he says; ‘‘ dabei nehmen wir vorliufig an, dass der vordere und der hintere Driisenblattkeim aus dem Ptychoblast und nicht aus dem Contributions to the Study of Silk-Worms. IOI Ectoderm des Stomo-und Proctodiums hervorgeht.” But according to his Fig. III. Taf. X., which closely corresponds to our Fig. 54 or 55, we might say that the entoderm arises out of the epithelium of the stomodium. Schwartze (‘98) made a very valuable observation on this point and comes to the following conclusion: ‘‘ Vorder-und Enddarm entste- hen als Ektodermeinstiilpungen, dass Mitteldarmepithel aus seitlichen Zelllamellen, die von den blinden Enden Vorder-und Enddarmes aus auf einander zuwachsen, bis sie sich jederseits in der Mitte treffen, und sich dann in Folge starken Breitenwachsthums erst ventral, dann dorsal in der Mediane vereinigen. Der Mitteldarm ist also, abgeselen von der mesodermalen Muscularis, wie Vorder-und Enddarm rein ektodermaler Natur.” From the accounts above given, we may come to the conclusion that the view maintained by Heymons and Schwartze as to the formation of the entoderm from the ectoderm is to be accepted, and that the opinions of Tichomiroff and others are untenable, at least in respect to the order of the Lepidoptera. } Among the Coleoptera, Kowalevsky ('71) was the first author who worked on Hydrophilus, which was investigated in a more detailed manner by Heider ('85, ‘89). Both these observers saw a rhomboidal area at the anterior end of the blastopore, which remained open after the closure of the other portion of the blastopore, thus corresponding exactly with what we saw in the silk-worm. In the sections passing through this portion, Heider distinguishes two cell-layers in the gastrula tube, namely ‘‘1. ein Paar von seitlichen Divertikeln, deren Hohlraum durch seitliches Auswachsen des Urdarmlumens hervorgegangen ist und 2. eine mediane an die paarigen Divertikeln sich dicht ausschliessende Zellmasse. Aus den Divertikeln werben die Mesodermmassen des Kopfsegments und des spateren Mandi- dularsegments, wihrend die unpaare Zellmasse zur Ectodermanlage (Entodermanlage ?) wird.” A precisely similar result was obtained by Wheeler (/89) in Doryphora, and it was also confirmed by Graber ('90) who says ‘“‘die Ptychoblastmetameren sind bei Lina alle mit Ausnahme der zwei polaren oder Endsegmente, das ist des procephalen und analen 102 K. Toyama: Abschnittes rein mesodermatische Bildungen, beziehungsweise Anlagen, wahrend die gennanten zwei Segmente gemischter Natur sind, dass heisst ausser dem auch ihnen zukommenden und sogar sehr stark entwickelten Mesodermantheil zugleich die Entodermanlage enthalten.” Voeltzkow ('89), on the other hand, observed quite correctly the formation of the entoderm from the cells of the stomodium and the procto- dium. He was followed by Lecaillon ('98) who worked on the formation of this layer in a leaf beetle. The subject has been quite recently taken up by Deegener (‘00) who made a study on Hydrophilus and came to the following conclusion : “Auch ich kann mich mit Kowalevsky und Heider’s Darstellung nicht einverstanden erklaren und stimme vielmehr mit Voeltzkow und Lecaillon iiberein. Beide Forscher finden, dass bei Melolontha bezw. einer Anzahl untersuchter Chrysomeliden der Ursprung des Mitteldarmepithels in zwei vorderen und zwei hinteren, vom Vorder- bezw. vom Enddarm auswach- senden ventrolateralen ektodermalen Lamellen zu suchen ist, die durch ihre Vereinigung in der Mitte, sowie in der ventralen und spater in der dorsalen Medianlinie das Mitteldarmrohr entstehen lassen. Das Wachs- thum der ektodermalen Lamellen geschieht tiberall ohne Beziehung der Zellen des unteren Blattes, so dass an dem ektodermalen Charakter das gesammten Mitteldarmepithels kein Zweifel herschen kann.” Comparing now the various accounts above referred to with the results above given respecting the formation of the entoderm in the silk-worm, we are strongly inclined to believe that in Coleoptera the entoderm arises much in the same way as in Lepidoptera and that the opinion advanced by Voeltzkow, Lecaillon and Deegener is to be maintained. The question naturally arises: What is the structure which was observed by Heider and Wheeler ? Now if we compare the Figs. 61, 62, 71, 76, and 77 given by Heider with our Figs, 2 and 2%, we shall at once be struck with the similarity of his entoderm to the oral cell-mass of the silk-worm. This similarity becomes more pronounced if we recollect that in Doryphora Wheeler saw the migration of the cells out of the structure which he calls the entoderm- centre and which corresponds to the entoderm of Heider in much the Contributions to the Study of Silk-Worms. 103 same way as the migration of the cells takes place out of the oral cell-mass in silk-worm. Wheeler says: ‘*Be this as it may, in later stages I believe it can be shown that cells do migrate into the yolk from the embryo and especially from the entoderm-centres. This was shown by me to be the case in Doryphora, where many cells pass into the yolk from either entoderm pole. I have since observed an exactly similar phenomena in Telea polyphemus! in a corresponding stage of develop- ment.” A similar phenomenon was also observed by Graber ('89), in Melolontha. Heymons considers these cells to be the ‘‘ Paracyten,” but we think that they are nothing else than the oral cell-mass. Tichomiroff ('92) still maintains the opinion: ‘‘Je puis confirmer ice que la participation des cellules vitellines dans la formation du Mésoderme et de |’epithelium de l’'intestin moyen est trés clairement vue sur les preparations de M™*. O. Tichomiroff (’90) chez la chrysopa et la pulex et sur mes propres preparations, de la Calandra granatia ('92).” Although he (‘92) has stated that there exists close relationship between the yolk-cells and the entoderm, and shows his Fig. 6 as a proof of it, yet there remains some doubt as to whether yolk-cells are changed into the entoderm, or indeed as to whether there exists any such relation- ship between them. In other orders of insects, the formation of the entoderm has been mostly studied in Orthoptera. Ayers ('84, in Oecanthus) advanced the opinion that it is derived from the yolk-cells, while a considerable number of observers derive it from the ento-mesoderm (Korotneff, '85, in Gryllotalpa; Nussbaum, '88, in Blatta; Cholodkowsky, ‘88, ‘g1, in Blatta ; Wheeler, '89, '93,* in Blatta etc). Heymons ('95) as before said, in his beautiful monograph asserts that the entoderm is derived from the stomodial or proctodial wall by cell-proliferation and concludes that ‘‘ Bei den von mir untersuchten (comparing six genera such as Forficula, Gryllus, Gryllotalpa, Periplanata, Phyllodromia, Eclobia) ist somit der ganze 1 The cells here are I think undoubtedly the oral cell-mass, * If we look at his Figs. 32, 33, and 34, and the description that “ median portion thus proliferated beyond limits of the ectoderm is the anterior or oral entoderm-centre,” it is clear that in Xyphidium the entoderm arises from the ectoderm by cell-proliferation, 104 K. Toyama: ’ Darmtraktus ausschliesslich ektodermaler Natur.” Rabito (‘98) also came to the same conclusion in Mantis. Concerning the oral cell-mass in Orthoptera as far as we are aware there exists no literature. Next to Orthoptera Muscidae have been much studied. It was in studies on this insect that Kowalevsky ('86) first advanced the theory of the ento-mesoderm, which in the hands of Heider, Wheeler, Graber et al. inaugulated a revolution in the opinions respecting entoderm- formation in insects. As early as 1889 Voeclotzkow, however, came to the conclusion that in Musca the entoderm is formed by the proliferation of the cells of the stomodial and proctodial epithelium. Graber (90) criticized this and came to the following opinion: ‘‘Zudem habe ich in meiner Muscidenarbeit héchst eigenthiimliche, bei anderen Insekten bisher véllig unbekannte Verhaltnisse nachgewiesen, nach denen es mindestens méglich ist, dass hier die Drtisenblattkeime zum Theile aus ganz selbstindigen Einstiil- pungen des Keimstreifenepithels aus den sogenannten lateralen Gastral- falten entstehen.” But in his later paper (91) he also maintains the ectodermal origin. Ritter (‘90) on the other hand, shows that in Chiro- nomus, the entoderm is separated off from the ‘‘segmentweise Wiilste” of the mesoderm (see his Fig. 30). After considering these results, Heymons ('95) expressed his view in the following words: ‘‘ Nach den bisherigen Untersuchungen zu urtheilen, ist es daher in hohen Grade unwahrscheinlich, dass das Mittledarmepithel der Musciden aus dem unteren Blatt (Entomesoderm) entsteht.” Quite recently Escherich (‘oo, ‘o1) has found three folds in the anterior portion of the blastopore of Musca, of which he writes ‘‘ sie stellen einen - Theil der Mesodermanlage dar; die mediane Falte dagegen ist, wie wir gleich sehen werden, die vordere Anlage des Entoderms. Wir haben also in diesem Stadium bei den Musciden in der That ganz ahnliche Verhiltnisse, wie bei Sagitta, worauf ja bekanntlich schon Biitschli und Kowalevsky hingewiesen haben.”’ And he finally came to the conclusion, ‘‘ dass I. bei den Musciden sich sehr frithzeitig cin Entoderm anlegt, dass 2. dieses 4 OS ie lt ed eg. rate Contributions to the Study of Silk-Worms, 105 Entoderm ein Abké6mmling des Blastoderms ist und dass 3. die Differenzie- rung der beiden primiren Keimblatter durch eine typische Invagination eingeleitet wird.” Further investigations are necessary before any decisive opinion on this point can be reached. Observations on the formation of the entoderm in other groups of insects hitherto considered are scanty, and the opinions formed by various authors vary greatly. Thus Patten ('84) claims that in Phryganids the entoderm arises from the yolk-cells, with which Will ('88) also agrees in his observation of viviparous Aphis. Witlaczil ('84), however, holds the view that in oviparous aphis the entoderm is derived from the ectoderm. Carriére (‘90) in his study on the carpenter-bee (Chalicodoma) has found a cell-thickening at the fore and hind portions of the midplate, and the cell-mass proliferated from the thickenings is converted into the entoderm- anlage, a fact which stands well with the view that the entoderm is formed out of the ectoderm. Kulagin’s observation (’98) on Platygaster differs entirely from the results obtained by all other authors hitherto mentioned. According to his observations the inner layer is formed by the immigration of the blastoderm cells of which he says as follows: ‘‘folglich entsteht das Entoderm und Mesoderm gleichzeitig auf dem Wege der Theilung der Zellen des Blastoderms und ihrer Einwanderung.” This seems to be the most rudimentary form of the entoderm-formation hitherto discovered, Now considering the facts and arguments thus brought out by various authors on different kinds of insects, the most usual mode of the formation of the entoderm in the class Insecta is that it arises from two separate centres—the oral andthe anal. But since the entoderm of many other animals arises froma single centre it is tacitly assumed that such must originally have been the case also with insects, and that the present bipolar condition must be due toa secondary modification. Starting with this postulate, Kowalevsky has formulated his hypothesis that ‘* bei der so in die Linge gezogenen Gastrula der Insekten der mittlere, das Entoderm liefernde sack so ausgezogen ist, das er in der Mitte ganz verschwindet und nur an seinem vorderen und hinteren Ende bestehen 106 K. Toyama: bleibt.” But if as we assume that the entoderm of Kowalevsky corresponds to our oral cell-mass which disintegrates into free cells, and that the definite entoderm is formed from the cell proliferation of the stomodium and the proctodium, then it will not be far from the truth, if we say that the original entoderm-anlage are entirely Jost in imsects, at least in the higher forms, and to supply this want ectoderm cells separate off at their original places and form the entoderm. Blood cells. The blood cells of Bombyx mori are said by Dohrn ('96) to have some relation to the yolk cells. Ayers ('84) also maintains the same view as Dohrn in his studies on Oecanthus. Will (‘88) comes to the same opinion in regard to Aphis saying that the cell-elements of the blood arise from entodermal yolk-cells, ‘‘sowhol innerhalb des Herzens als auch frei in der Leibeshdhle.” Cholodkowsky (‘91) also holds the same opinion respecting Phyllodromia. Korotneff ('83, '85) on the other hand states that in Gryllotalpa at an early period blood cells are found almost everywhere between the yolk and the mesoderm; they are derived, as he states, from the cells of the somatic mesoderm layer which lost their connection with the other parts of the mesoderm, and have fallen into the body cavity. Patten ('84) assumes a similar view in his researches on Phryganids. Wheeler (89) again confirms the view that the endoderm is formed out of the mesoderm in Doryphora, though he differs somewhat from the above two observers as to the place of its origin. Heymon’s statement ('95) much resembles that of Korotneff. He says that ‘bei Forficula, sowie bei den ‘‘hier betrachteten Blattiden und Grylliden sind die Blut- kérperchen mesodermaler Abkunft. Sie entstehen aus Mesodermzellen, welche nicht bei der Bildung der Ursegmente sich betheiligt hatten, sondern zwischen diesen in der Medianlinie des K6rpers ihren Platz beibehalten.” A similar view is held by Lécaillon (‘98) in the case of Chrysomelidae, and by Schwartze in the case of Lasiocampa. But in the case of Lasiocampa, the last author states that ‘‘ diese Einwanderung é ad Contributions to the Study of Silk-Worms. 107 beschrinkt ist auf diejenige Stelle des Embryos, an die sich die Ektoderm- rinne von zuletzt schliesst, d.h. auf einen sehr geringen Theil der ganzen Liinge des Keimstreifs,” and further on, in other portions of the mesoderm, he says, that these ‘‘ zeigen niemals eine Lockerung ihrer Zellen.” It seems to us that he has considered that portion of the embryo which corresponds to our oral cell-mass as the only structure which forms the blood cells. From all the opinions above considered those of Korotneff and Heymons arein best accord with the formation of the blood corpuscles as we have observed it in the silk-worm and as it is described in previous pages, and we must believe that the various views held by the other authors are to be looked upon as being due to their having confused the various cell elements found in the yolk. First of July, 1go1. Zoological Institute, College of Agriculture, Tokyo Imperial University. 108 K. Toyama: Works Referred to. Ayers, H. ('84), On the Development of Oecanthus niveus and its parasite Teleas. Mem. Boston Soc. Nat. Hist., Vol. 3, 1884. Bobretzky, N. (78), Uber die Bildung des Blastoderms und der Keimblatter bei den Insekten. Zeitschr. wiss. Zoologie. Bd. 31, 1878. Boas, J. (99), Einige Bemerkungen tiber die Metamorphose der Insekten. Zool. Jahrf. Abth. f. Syst. Geogr. and Biol. d. Thiere. Bd. XII. 18a9. Bruce, A. T. ('87), Observations on the Embryology of Insects and Arachnids. A memorial volume, Baltimore, 1887. Carriere, J. ('90), Zur Embryonalentwicklung der Mauerbiene (Chalico- doma muraria, Fabr.). Zool. Anzeiger, Bd. 13, 1890. (‘90), Die Entwickelung der Mauerbiene (Chalicodoma muraria) im Ei. Arch. f. mik. Anatomie, Bd. 35, 1890. (‘90), Die Driisen am ersten Hinterleibsringe der Insektenembryo- nen. Biol. Centralblatt, 18or. Cholodkowsky, N. ('88), Uber die Bildung des Entoderms bei Blatta germa- nica. Zool. Anzeiger, Jahrg. 11, 1888. (‘90), Zur Embryologie von Blatta germanica. Zool. Anzeiger, Jahrg. 13, 1890. *('o1), Die Embryonalentwicklung von Phylodromia germanica. Mem. Acad. St. Pétersbourg. Tom. 38, 1891. Deegener, P. (‘oo), Entwicklung der Mundwerkzeuge und des Darmkanals von Hydrophilus. Zeitschr. f. wiss. Zoologie, Bd. 68, 190. Dohrn, A. ('76), Notizen zur Kenntniss der Insektenentwicklung. Zeitschr. f. wiss. Zoologie, Bd. 26, 1876. (‘or), Entwicklung der Mundwerkzeuge und des Darmkanals von Hydrophilus. Biol. Centralblatt, Bd. XXII. rgor. Escherich, K. ('00), Uber die Bildung der Keimblitter bei den Musciden. Nov. Acta. Leop. Carol. Bd. LXXVII., 1900. (‘o1), Das Insekten-Entoderm. Biol. Centralblatt, Bd, XXI., 1901. Graber, V. ('88), Vergleichende Studien iiber die Keimhiillen und die Contributions to the Study of Silk-Worms. 109 Riickenbildung der Insekten. Denkschr. Acad. Wiss. Wien, Bd. 14, 1888. *('89), Vergleichende Studien tiber Die Embryologie der Insekten und insbesondere der Musciden. Denkschr. Acad. Wiss. Wien, Bd. 56, 1889. (‘c0), Vergleichende Studien am Keimstreifen der Insekten. Denkschr. Acad. Wiss. Wien, Bd. 57, 1890. (‘o1), Bemerkungen zu J. Carriere’s Aufstaz ,, Die Driisen am ersten Hinterleibsringe.” Biol. Centralblatt. 1891. *(‘g1), Beitrage zur vergleichende Embryologie der Insekten. Denkschr, Acad. Wiss, Wien, Bd. 58, 1891. ('91), Uber die embryonale Anlage des Blut-und Fettgewebe der Insekten. Biol. Centralblatt, Bd. rr. Hatschek, B. ('77), Beitrige zur Entwicklungsgeschichte der Lepidop- teren. Jenaische Zeitschr, f. Naturw., Bd. 11, 1877. Heider, K. ('85), Uber die Anlage der Keimblatter von Hydrophilus piceus, L. Abhandl, d. k. preuss. Acad. Wiss. Berlin, 1885. ——— (‘89), Die Embryonalentwicklung von Hydrophilus piceus, L. I. Theil. Jena, 1889. Heymons, R. ('95), Die Segmentirung des Insektenkérpers, aus dem Anhang z.d. Abhandlungen der Ko6nigl. preuss. Akademie d. Wiss. z. Berlin, 1895. ('95), Die Embryonalentwicklung von Dermapteren und Ortho- pteren, unter besonderer Beriicksichtigung der Keimblatterbildung, Jena, 1895. ('95), Grundziige der Entwicklung und des Korperbaues von Odonaten und Ephemeriden, aus dem Anhang zu den Abhandlungen d. KGnig]l. preuss. Akademie d. Wiss. z. Berlin, 1896. Kowalevsky, A. ('71), Embryologische Studien an Wiirmern und Arthropo- Sef Mem. Acad. imp, sscienc. St. ,Petersbourg, Sér. 7, Tom. 16, No. 12, 1871. (‘86), Zur embryonalen Entwicklung der Musciden. Biol. Central- blatt, 6 Bd. 1886, 1IO K. Toyama: Korotneff, A. (85), Die Embryologie der Grytlota!pa, Zeitschr. f. wiss. Zoologie, Bd. 41, 1885. ('94), Zur Entwicklung des Mitteldarmes bei den Arthropoden. Bic! Centralblatt, Bd. XIV., 1894. Koschevnikov, G. A. (‘0o), Ueber den Fettkérper und die Oenocyten der Honigbiene. Zool. Anzg., 1900. Krancher, 0. ('81), Der Bau der Stigmen bei den Insekten. Zeitschr. f. wiss. Zoologie. Bd. XXV., 1881. Kulagin, (98), Beitrage zur Kenntniss der Entwicklungsgeschichte von Platygaster. Zeitschr. f. wiss. Zoologie, Bd. LXIII., 1898. Korschelt und Heider, ('92), Lehrbuch der vergleichenden Entwicklungs- geschichte der wirbellosen Thiere. Heft, 2, Jena, 1892. Lecaillon ('98), Recherches sur l’oeuf et sur le developpement embryonaire de quelques Chrysomelides. Pairs, 1898. Mayer, P. ('76), Uber Ontogenie und Phylogenie der Insekten. Jen. Zeischer. f, Naturwiss. Bd. 10, 1876. Packard, A. ('93), A study of the transformations and anatomy of Lagoa crispata. 1893. — ('98), A Text-book of Entomology. New York. 1898. Patten, W. ('84), The development of Phryganids with a preliminary note on the development of Blatta germanica. Quart. Jour. Micr. Science, vol. 24, 1884. Ritter, R. (90), Die Entwicklung der Geschlechtsorgane und des Darmes bei Chironomus. Zeitschr. f. wiss. Zoologie, Bd. 50, 1890. Rabito, (‘98), Sullorigine dell’intestins medio nella Mantis religiosa. Palerms, 1808. Schwartze, (‘99), Zur Kenntniss der Darmentwicklung bei Lepidopteren. Zeitschr, f. wiss. Zoologie. Bd. 65, 1899. Tichomiroff, A. (‘91), Dévelopment du vera soie du murier dans l’oeuf. Rapport présenté a la chambre de commerce de Lyon. 1891. ('79), Uber die Entwicklungsgeschichte des Seidenwurms. Zool. Anzeiger, Jahrg. 2, Nr. 20. (‘92), Aus der Entwicklungesgeschichte der Insekten. Festschrift zum 70. Geburtstage Rudolf Leuckarts, Leipzig, 1892. Contributions to the Study of Silk-Worms. III Tichomirowa, 0. (’co), Zur Embryologie von Chrysopa. Biol. Centralblatt, Bd. 10, 1890. Uzel, H. (’97), Vorlaufige Mittheilung tiber die Entwicklung der Thysa- nuren. Biol. Centralblatt, Bd. XX., 1897. Will, L. ('88), Entwicklungsgeschichte der viviparen Aphiden. Zool. Jahrb. Abth. f. Anatomie und Ontogenie, Bd. 3. 1888. Witlaczil, E. ('84), Entwicklungsgeschichte der Aphiden, Zeitschr. f. wiss. Zoologie, Bd. 40, 1884. Wheeler, W. M. ('89), The embryology of Blatta germanica and Doryphora decemlineata. Jour. of Morphology, vol. 3, 1889. ('‘93), A contribution to Insect Embryology. Jour. of Morphology, Vol. 8, 1893. - Voeltzkow, A. ('89),* Entwicklung im Ei von Musca vomitoria. Arbeit. Zool. Zoot. Inst. Wiirzburg, Bd. 9, 18809. (‘89),* Melolontha vulgaris. Ein Beitrag zur Entwicklung im Ei bei Insekten. Arbeit. Zool. Zoot. Inst. Wurzburg, Bd. 9, 1889. Schaffer, C. ('89), Beitrage zur Histologie der Insekten. Zool. Jahrb., Abth. f. Anatomie u. Ontogenie, Bd. 3, 1880. Verson, E, Vison, ('91), Cellule glandulari ipostigmatische. Padova, 18o1. Wielowiejski, H. v. ('86), Uber das Blutgewebe der Insekten. Zeischr. f. wiss. Zoologie, 1886. . | Selvatico ('82), Sullo soilippo embrionale dei Bombicini. Annuarid della r. stazione bacologica di Padova, 1882. * The asterisk marks the cases in which I have not been able to gain access to the original paper. I12 K. Toyama: Explanation of Plate I. Fig. 1. Surface view of embryo with blastopore not yet closed (taken from an egg, one month old: 28th, August.) Zeiss Ax4. 0, anterior widening of blastopore ; pcl, procephalic lobe ; bl, blastopore ; a, posterior widening of blastopore ; cd, anal segment. Figs. 2—5. Cross sections taken from embryo as in preceding figure. Zeiss Dx4q. Fig. 2. Section through primary head segment; knife passing through anterior portion of blasto- porous widening (Fig. I. 0). ect, ectoderm ; inv, cavity of invaginated gastrula ; vit, vitellophags ; yl, yolk-granules. Fig. 3. Section through posterior portion of primary head segment. ect, ectoderm; ms’, cell-mass formed by coalescence of invaginated gastrula; other letters as in preceding figures. Fig. 4. Section through maxillary segment. p.f, primitive furrow ; ms, mesoderm, Fig. 5. Section through posterior blastoporous widening, knife passing through at (a) Fig. I, showing lateral overgrowth of ectoderm to form mesoderm. am, ammion; other letterings as in preceding figures. Fig. 5.1 Section of anal segment. Letters as in preceding figures, Vig. 2.7 Transverse section through anterior widening of blastopore already closed (taken from an embryo somewhat older than Fig. I), am, amnion ; ect, ectoderm; o, ectodermal depression, par, “paracyten,” other letters as in preceding figures, Fig, 2.> Longitudinal section of embryo at same stage as in above figure. Lettering as in the preceding. Fig. 6. Surface view of embryo with blastopore about to close. At (a) it remains open (taken from the egg on November 30th.) Zeiss A xq. o1, ectodermal depression formed after closure of anterior widening (Fig. I. 0) of blastopore. Figs. 7—11. Cross sections taken from embryo as above figured, Zeiss D x 4. Fig. 7. Section through ectodermal depression (Fig. 6.0’) at primary head segment. am., ammion ; ect, ectoderm; 01, ectodermal depression ; ms, mesoderm; ms, cell-mass sprung from invaginated gastrula or oral cell-mass ; ms?, detached cell from oral cell-mass, remaining letters as in the preceding figures, Fig. 8. Section through middle portion of embryo. Lettering as in preceding figures. Fig. 9. Section through a portion, little posterior to the preceding figure. Lettering as in preceding figures. Fig. 10. Section through blastopore at Fig. 6.a., showing lateral overgrowth of ectoderm to form mesoderm or inner layer, ‘ » if, z a Ss Contributions to the Study of Silk-Worm. 113 Fig. rr, Section of anal segment. Lettering as in preceding figures, *Fig. 7.% Portion of same section, highly magnified, showing two migratory cells from oral cell-mass. (Zeiss Ap. 0. 2 mm x Comp. oc. 6.) Lettering as in the preceding figures. Fig. 12. Surface view of embryo after closure of blastopore, (taken from egg on February 2nd.) (Zeiss A x 4.) p.cl, procephalic lobe ; ed, anal segment. Figs. 13—16, cross sections of embryo of same stage as that represented in preceding figures. (Zeiss D x 4.) Fig. 13. Section of procephalic lobe at region of oral cell-mass, am, amnion ; ect, ectoderm; 0}, ectodermal depression at primary head segment ; ms1, oral cell-mass ; ms?, detached cells from the oral cell-mass ; d.c, degenerating cells ; vit, vitellophags. Fig. 13.7 Portion of same section more highly magnified, showing degenerating cells and vitellophags. (Zeiss Ap, 0. 2 mmx Con)p. oc. 4.) Lettering as in preceding figures, Fig. 14. Section through thoracic region, Lettering as in preceding figures. Fig. 15. Section through intersegmental region at the thoracic portion. Lettering as in the preceding. Fig. 16. Section through caudal plate, showing large accumulation of mesoderm, Lettering as in preceding, Fig. 17. Median longitudinal section of embryo, taken out on March 18th. (Zeiss Dx 4.) Lettering as in the preceding. Fig. 18. Surface view of an elongated embryo, taken out on March 27th. Lettering as in the preceding figures. Figs, 12—-27, consecutive series of sections at anterior portion of embryo, as shown in preceding figure. (Zeiss D x 4.) Figs. 19—25. Consecutive sections through primary head segment, showing relation of oral cell- mass to ectoderm. ect, ectoderm; am, amnion ; ms, mesoderm; ms}, oral cell-mass; ms?, detached oral cell-mass; b.c1, migrating mesodermal cells; par., ‘‘paracyten”’ or degenerating cells ; other letters as in preceding figures. Explanation of Plate VIII. Figs. 23--25. Sections through primary head segment, explanation as given above. Fig. 26, Section passing through first abdominal segment, (Zeiss Dx 5.) am, amnion ; n.f, neural furrow; ect, ectoderm ; ms%, “ paradermalen Schicht’’; ms?, “ paralecithalen Schicht”; vit, vitellophags. Fig. 27. Transverse section through procephalic lobe, showing migratory cells from oral cell-mass. 114 K. Toyama: am, amnion; ect, ectoderm; ms, mesoderm; ms?, oral cell-mass ; ms?, detached cells from oral cell-mass. Fig. 28. Portion of same section on left side, more highly magnified, showing migratory cells from oral cell-mass. (Zeiss Ap. o. 2 mm x Comp. oc. 6.) ms}, oral cell-mass ; ms?, detached cells from oral cell-mass ; yl, yolk granules. Fig. 29. Median longitudinal section of embryo at same stage as that represented in Fig. 18, which shows faint depression of stomodium at st. A, anterior ; P, posterior direction ; st, stomodium ; ms, mesoderm ; ms}, oral cell-mass ; ms?2, detached cells from oral cell-mass ; dc, degenerating cells ; vit, vitellophags. Fig. 30. Surface view of embryo taken out on March 27th. Ib first, sccond and third thoracic legs; n.f, neural furrow ; cd, anal segment. (Zeiss A x 4.) , labrum ; at, antenna; md, mandible; mx1~2, first and second maxillae; th. 11-3. Fig. 31. Sagittal section of same embryo as that represented in the above, skowing relation between stomodium and oral cell-mass, Lettering as in preceding figures. Fig. 32. Sagittal section of head segments of embryo more advanced than above. Ilere oral cell- mass is entirely lost. Lettering as in the preceding figures. Figs. 33—40. Series of sagittal sections of embryo at same stages as shown in Fig, 30, showing stomodial depression. Among these series, Fig. 40 is a section through the median longitudinal line. (Zeiss D x 4.) st, stomodium; ms, mesoderm; ms, detached cells from oral cell-mass ; a, elongation of lateral portion of stomodium, or ‘“ vordere Epithellamelle,” ms”, anlage of suboesophageal body, from the anterior portion of which (a) blood cells are formed, A, anterior ; P, posterior direction. Figs. 41—s5o. See explanation of Plate III. Arabic numerals placed between every two sections indicate the number of sections that intervene between the two (exclusive,) Explanation of Plate IX. Figs. 41—50. Series of transverse sections through primary head segment of embryo as in Fig. 30, showing deepening of stomodium, (Zeiss D x 4.) ect, ectoderm; ms, mesoderm; d.c, degenerating cells; st, stomodium; a, lateral prolongation of stomodial wall to form mid-gut ; b.c, blood cells. Other letters as in preceding, Vig. 51. Surface view of shortened embryo, taken from egg in April (Zeiss A x 4.) Ib, labrum ; at, antenna; md, mandible; mx.(1-2), first and second maxilla: th, 1,4-3), first, second and third thoracic legs ; sl.g, opening of silk gland ; ab, (4-19), abdominal “ legs ; stg, stigma ; n.f, neural furrow. (Zeiss A x 4.) Contributions to the Study of Silk-Worms. 115 Fig. 52. Transverse section through antennal region of same embryoas that represented in Fig. 5 showing entoderm-anlage. ent,? entoderm anlage or lateral prolongation of stcmodial wall; n, nerve. Other letters as in preceding figures, Fig. 53. Sagittal section through primary head segment of same embryo as in prece ling figure, sho v- ing stomodial tube and entoderm-anlage. ep]. v, ‘‘vordere Epithellamelle”’; st, stomodium ; ms, mesoderm ; g.l, ‘‘ Grenzlamella”’; s.b, suaboesophageal body ; b.c, blood cells. Fig. 54. Portion of same series of above section more highly magnified, showing blood cells. (Zeiss, Ap. 2. mm x Comp. oc. 6). ect, ectoderm of ‘“ Vorkiefersegment;” b.c, blood cells ; a, blood cells taken from dorsal vessel of full grown embryo about to hatch, Fig. 55. Sagittal section of embryo, somewhat more advanced than that represented in Fig. 52 (taken from egg on April, 4th. Zeiss D x 4). b.c, blood cells; ms, mesoderm ; st, stomodium ; g.l, “ Grenzlamelle.” epl. v, “ vordere Epithellamelle”’; s.b, suboesophageal body ; g.m, mandibular ganglion ; g.mx('~*), first and second maxillary ganglion ; sl.g, silk gland. Figs. 56—63. Series of transverse sections of embryo corresponding to preceding figure. (Zeiss, 1D) Se Iie Vigs. 56—57, sections through antennal region ; Figs. 58 —60, sections through man- dibular segment; Figs. 61—62, sections through first maxillary segment; Fig. 63, section through mesothoracic segment. at, antenna ; g.l, “Grenzlamelle’’; st, stomo- dium ; ent, lateral entoderm-anlage ; s.b, suboesophageal body ; b.c, blood cells; nc, nerve cord; md, mandible; ms1, first maxilla; n.f, neural furrow ; ms, mesoderm ; sp. ms, splanchnic mesoderm ; sm. ms, somatic mesoderm ; cce, coelomic cavity ; tent?, second tentorium ; d.c, degenerating cells; th. 12, second thoracic leg ; ce, cenocytes. Fig. 64. Transverse section through sixth abdominal segment, same embryo as above. (Zeiss Dx 4). ent, entoderm; d.c, degenerating cells ; g, genital cells ; sd. ms, splanchnic mesoderm ; stg, stigma. Fig. 65. Portion of frontal section through anterior portion of mid-gut showing karyokinetic division of ils epithelial cells. epl. v, “vordere Epithellamelle ”; g.l, ‘Grenzlamelle’’; sp. ms, splanchnic mesoderm ; st, stomodium, (Zeiss, Ap, 2. mm x oc. II). Fig. 66. Portion of transverse section of primary head segment of embryo, corresponding to that represented in Fig. 52. (Zeiss, Ap. 2. mm x Comp. oc. 4). ect, ectoderm ; par, degenerating cells. Figs. 67-—69. Free cells in yolk, taken from same embryo as in preceding figure (magnification as above). d.c, degenerating cells ; ms’, immigrating cells from oral-cell-mass ; vit, vitellophags ; > yl, yolk granules, 116 K. Toyama: Fig. 68. Yolk ball, taken from embryo as represented in Fig. I, showing direct division of nucleus, Fig. 70. Anal segment of embryo as represented in Fig. 30. (Zeiss, Bx 2). n.f, neural furrow ; a, proctodial depression. Fig. 73. Transverse section through anal segment of embryo as represented in Fig, 30, showing bifurcation of neural furrow, (Zeiss D x 4). nis, mesoderm ; n.f, neural furrow. Fig. 76. Transverse section through anterior portion of proctodium of embryo corresponding to Fig: Chil, (VASE IDS 30)). pt, proctodium ; m.v, malpighian vessel ; ms, mesodeim; n.c, nerve cord. Explanation of Plate X. Fig. 71. Longitudinal section of same embryo as Fig. 70. (Zeiss Dx 4). d.c, degenerating cells; ms, mesoderm ; pt, proctodium. Fig. 72. Sagittal section through anal segment of an embryo slightly more advanced than the above. (Zeiss, D x 4). ent, entoderm-anlage ; m.v, malpighian vessels; pt, proctodium; other letters as in preceding figures. Fig. 73. Sagittal section through posterior portion of embryo represented in Fig, 51. (from embryo taken on April 1st). (Zeiss, Dx 4). am, amnion cells; d.c, degenerating cells; epl. h, “hintere Epithellamelle”; pt, procto- dium. Fig. 75. Sagittal section through abdominal portion of more advanced embryo, taken from egg on April gth. (Zeiss D x 4). Lettering as in preceding figures, Pig. 77—82. Series of transverse section through head segment of embryo as represented in Fig. 52, showing various ectodermal invaginations, (Zeiss, Dx 4). Figs. 77, section through anterior portion of mandibular segment in somewhat oblique direction ; Fig. 78, posterior portion of same segment ; Fig, 79, first maxillary segment; Fig. 80—82, series of sections through second maxillary segment, at, antenna; br, supraoesophageal ganglion ; ent, entoderm; f.n.c, frontal nerve chord; fl. md, flexsor mandibulae ; s.b, suboesophageal body ; sp. ms, splanchnic mesoderm ; sl.g, silk gland ; md, mandible; mx (1-2), first and second maxilla; mn, new gland or hypostigmatic gland ; st, stomodium. Explanation of Plate XI. Figs. 83—91. Consecutive series of transverse sections through head of embryo more advanced than that represented in Fig, 52, (taken from egg on April 6th). (Zeiss D x 4). Figs. 83 and 84, sections through anterior portion of mandibular region; Fig. 85 through posterior portion of same region; Figs. 86 and 87, through portion between mandible and first maxilla ; Figs, 88 and 89, through second maxilla; Fig. 90, through posterior Contributions to the Study of Silk-Worms. ney portion of head ; Fig, 91, through first thoracic segment, br, supraoesophageal ganglion; ex. md, attachment of extensor mandibulae; fl. md, attachment of flexor mandibulae ; g.f, ganglion frontale; n, new gland or hypostigmatic gland; s.g, salivary gland; st, stomodium ; sl.g, silk gland; tent(1=?), first and second tentorium; other letters as in preceding figures. Figs. 92—-93. Series of sagittal sections through head and thorax of embryo as in preceding figure. (Zeiss, D x 2). Lettering as in preceding figures, Fig. 94. Frontal section through head and thorax of more advanced embryo taken out on April gth. (Zeiss, Dx 4). Lettering as in preceding figures. Fig. 95. New gland at base of first stigma, (taken from larva of third stage). (Zeiss, A x 2). g.a, abdominal ganglia ; m, muscles; n, new gland or hypostigmatic gland ; tr, trachea. Fig. 96. New gland of full grown larva. (Zeiss, Ax 2). Figs. 67(1-2), Portion of the new gland, highly magnified. (Zeiss D x 2). Arabic numerals placed between every two sections indicate the number of sections that intervene between the two (exclusive). CONTENTS. Introductory. Methods. ... A. My own observations. I. The formation of mesoderm and entoderm. The formation of the mid-gut. The proctodium and the Malpighian vessels. ... II. The vitellophags and other cellular elements found in the yolk. a. Vitellophags. 6. Migratory cells from the ectoderm. c. The cells migrating from the oral cell-mass. d. The blood cells. e. Degenerating cells. ... III. The endoskeleton of the head, with reference to the salivery gland and a new gland. B. General considerations. The mesoderm... The entoderm ... Blood-cells. Works referred to. ... Explanation of plates. Contents. ... PAGE. & Auctor del. 1 BULL. AGRIC. COLL. VOL. V. rts Ae wa! . J nah t Auctor del, PLATE Vil. : BULL. AGRIC. COLL. VOL. V. eset? ee oo mo 3 s 6 S Oo e----- “~ Auctor del. ILL. AGRIC. COLL. VOL. V. PLATE X. \ 1 i f ¥ ; ' ‘ " n t \ ’ ' ‘ : . “ i \ . ¢ , I ‘ . ‘ . { ; f . ° . ‘ te td , “ ; ~ a * \¥ , 2 ‘ ‘ . . ) to 4 »* te ‘es BULL. AGRIC. COLL. VOL. V. It bo Auctor del. PLATE XT. 479 = ~~ } Ueber das wirksame Princip des Tuberculinum Kochii. VON N. Nitta. Einleitung und Literatur. Seit R. Koch sein Tuberculinpraeparat in Anwendung brachte, haben viele Forscher versucht, das wirksame Princip desselben zu isoliren. Koch selbst hat sein sogenanntes Reintuberculin aus dem Rohtuberculin oder gewohnlichen Tuberculin mittelst 60% igen Alcohol dargestellt und zeigte, dass es im Wesentlichen die chemischen Eigenschaften einer Albumose besitzt. Nach W. Kiihne ist dieses Produkt aber kein chemisches Indivi- duum, sondern ein Gemisch von Proteinstoffen, das noch an 20% Aschen- bestandtheile enthalt. Aus Tuberkelbacillenculturen hat W. Kiihne folgende verschiedene Proteinfractionen erhalten: Albuminat, Acroalbu- mose und Deuteroalbumone ; Essigsdurefallung und Ammonsulfatfallung, ! von denen einige beim Tierversuche eine noch energischere Wirkuug als das ,, Reintuberculin” Koch's zeigten. Trotzdem ist er der Meinung ,,dass keine dieser Substanzen mehr sei, als der Trager des Tuberculinum verum ; sie sind dazu unter sich in chemischen Verhalten zu verschieden und uns aus dem Nahrboden und als Bestandtheile des Handelspeptons nur allzu bekannt.” Auch hat Helmann eine eingehende chemische und pharmacologische Untersuchung .iiber das Tuberculin ausgefiihrt. Er fand als dessen Bestandtheile: 1) Albumose und zwar Protoalbumose und Deutero- albumose, 2) mehrere Alkaloide, 3) Extractivstoffe, 4) Mucin, anorganische Salze, Glycerin und Farbstoffe. Ferner stellte er mit dem Alcohol- niederschlag des Tuberculins(A) und mit dem Alcoholfiltrat (C) Tierversuche 1 Letztere beide Fractionen wurden aus albumosefreiag Peptoncultur erhalten, 120 N. Nitta: an und fand, dass ersteres, welches den gréssten Teil der Albumose enthalt, starke, entziindliche Local-Reaction und kein oder nur geringes Fieber, dagegen letzteres, welches besonders die Salze und nur wenig Albumosen enthalt, keine locale Entziindung, aber hohes Fieber ver- ursacht. Helman hat tiber das von ihm aus Kartoffelkulturen, welche unter oder ohne Mitwirkung von Serum und Glycerin gewachsen waren, erhaltene Tuberculin, sowie tiber die Koch’sche Lymphe Studien ver6ffent- licht. Er sieht sich auf Grund seiner Beobachtungen zu dem Schluss gezwungen, dass der wirksame Korper nicht aus Albumosen allein besteht; das ,,Reintuberculin” von Koch ist seiner Meinung nach eine Mischung von Albumosen und wirksamer Substanz; das Eiweiss reisse beim Aus- fallen gewisse in der Fliissigkeit enthaltene Substanzen mechanisch mit nieder. Matthes bekam durch Injection von gew6hnlichen Albumosen, also von K6rpern, welche ohne jede specifische bacterielle Thatigkeit aus Verdau- ungsgemischen isolirt werden, bei an Lupus erkrankten Menschen deutliche locale Reaction und halt die Tuberculinwirkuug wenigstens zum Theil fiir eine Wirkung einer gewohnlichen Albumose: daher empfiehlt er statt des Tuberculins, welches nach ihm ein theures, schwer haltbares Praeparat und noch dazu kein einheitlicher K6érper ist, die gew6hnliche Deuteroalbumose zu benutzen, welche allerdings in etwas grésseren Dosen (0,05-0,075 g) angewendet werden muss. Die Deuteroalbumose ist ferner ein vollig reines Material, welches sich leicht vollkommen salzfrei darstell- en lisst; sie erhalt sich als weisses trockenes Pulver jahrelang unverindert und erlaubt eine absolut genaue Dosirung. Petri und Maassen zeigten, dass gew6dhnliche 10 proc. Pepton- bouillon, welche fiir gesunde Meerschweinchen in Mengen von 4 ccm. eingespritzt ohne Nachtheil war, tuberculése Tiere tétete. Bei der Section fanden sich in der Umgebung der tnberculésen Herde deutliche Erschei- nungen einer Reaction. Nach Ansicht der Verff. ist diese Giftwirkung wesentlich dem hohen Peptongehalt der Nahrbouillon zuzuschreiben ; sie laugnen deshalb die Specifitaet der Tuberculinwirkung, womit auch mehrere andere Autoren tibereinstimmen. Von Stoffen, die dem Tuberculin ahnlich wirken, werden genannt: Ueber das wirksame Princip des Taberculinum Kochii. I2I ein proteinhaltiges Extrakt des Bac. pyocyaneus (Roemer), das Protein der Pneumobacillen oder des Bac. prodigiosus (Buchner), Proteine nicht pathogener Bakterienarten (G. Klemperer), Teucrin, ein Pflanzenextrakt (v. Mosetig), Kreatin, Kreatinin, Cystin, Allantoin und Tyrosin (Dixon und Zuill). Ausserdem noch eine grosse Anzahl der verschiedenartigsten Stoffe wie Thiophen, Benzol, Sulfoharnstoff, Sulfoathylharnstoff, Aceton, Propylamin, Trimethylamin, Allylamin, Taurin, Cadaverin (Spiegler), _ kantharidinsaure Salze (Liebreich) u. s. w. Nach der Untersuchung Viquerat’s verhalt sich Tuberculin, welches auf 150-200° c. erhitzt wurde, gegen tuberculése Tiere wie nicht erhitztes ; seine weitere Behauptung, dass bernsteinsaure Salze ebenso wirken wie Tuberculin, wurde von Hutyra sowie von mir selbst nicht bestatigt gefunden. Ruppeli untersuchte Filtrate von Massenculturen des Tuberkelbacillus auf ihre chemischen Bestandtheile. Die Filtrate enthielten an fallbaren Substanzen vorwiegend Deuteroalbumosen, neben primaren Albumosen und Hemialbumosen ; Acroalbumose ist nur dem Gehalt an Witte’s Pepton entsprechend wenig vorhanden. Es gelang nicht, ein typisches und specifisches Stoffwechselprodukt des Tuberkelbacillus zu isoliren; es liess sich nur feststellen, dass den Tuberkelbacillen ein tryptisches Verdau- ungsverm6gen zukommt, indem sie in alkalischer Lésung Eiweisskérper bis zur Bildung von Pepton unter gleichzeitigem Auftreten von Tryptophan spalten. Bei der grossen Divergenz der Meinungen verschiedener Forscher waren weitere eingehende Untersuchungen wiinschenswerth, und ich habe deshalb viele Versuche angestellt, um einen kleinen Beitrag zur Auf- klarung der Frage zu liefern. Eigene Untersuchungen. I. Herstellung des Tuberculins. Das bei meinen vergleichenden Versuchen angewandte Koch’sche Rohtuberculin wurde von mir in meinen Laboratorium aus Menschen- 122 N. Nitta: tuberkelbacillenkulturen hergestellt. Die Nahrlésung bestand aus einer normalen Peptonrindfleischbouillon, die eine 1.6-3.39§ Normalnatronlauge entsprechende Aciditat besitzt und die ich mit 5% Glycerin versetzte. Die Kultur wurde im Brutofen bei einer Temperatur von 37-39° C. gehalten bis nach Ablauf von 1-2 Monaten die Entwicklung der Bacillen beendigt war, worauf im Dampftopf 3-1 Stunde lang sterilisirt und durch sterilisirtes Filtrirpapier filtrirt wurde: das Filtrat wurde auf dem Wasserbade bei 93-100° C. in sterilisirter Porzellanschale bis auf den zehnten Theil ihres urspriinglichen Gewichts eingedampft. Die in dieser Weise gewonnene gelbbraune, syrupartige Flissigkeit stellt das sogenannte Tuberculin (Rohtuberculin, gewohnliches Tuberculin) dar. Sie reagirte schwach alkalisch und zeigte das spec. Gewicht 1.195 (bei 15° C.). Die Analyse ergab: Wasser 44.637599. Stickstoff 2.4750%. Asche 6.33229. Chlor 2.8609%. Das Glycerin wurde nicht bestimmt. Tuberculés gemachte Meerschweinchen zeigten nach Injection von 0,001 ccm. starkes Reactions- fieber (1-2° und dariiber): bei hochgradig tuberculésen Tieren (4-8 Wochen nach der Impfung) geniigt 0,1 ccm. Tuberculin zur Totung inner- halb 6-30 Stunden. Die Lethaldose fiir die Tiere mit weniger fortgeschrit- tener Tuberculose ist 0,25-0,5 ccm. II. Der mit 60% igem Alcohol erhaltene Niederschlag, das Reintuberculin Koch’s. Ein Teil des fliissigen Rohtuberculins wird mit nur 13 Volumenteilen absoluten Alcohols vermengt und 24-48 Stunden stehen gelassen: es bildet sich ein flockiger Bodensatz; die tiberstehende Flissigkeit wird abgegossen, nun aber ein gleiches Volum nur 6096 ige Alcohol zugesetzt, wieder absitzen gelassen und dies 3-4 Mal wiederholt, bis der tiber dem Niederschlag stehende Alcohol fast farblos erscheint: nach mehrmaligem Auswaschen mit absolutem Alcohol wird der Niederschlag bei 50=60° C. getrocknet, wobei eine schneeweisse leicht zerreibliche Masse resultirt. Dies ist das sogenannte Koch’sche Reintuberculin (Tuberculinum depuratum Koch). Tuberculése Meerschweinchen reagiren auf Injection von 0,00005 g. dieses Praeparats. Ueber das wirksame Princip des Tuberculinum Kochii. 123 III. Der in absolutem Alcohol unlosliche Teil des Rohtuberculins. Anfangs modificirte ich die Koch’sche Methode auf die Weise, dass ich, statt sechzigprocentigem, absoluten Alcohol anwandte. Wird das Rohtuberculin mit dem mehrfachen Volumen absoluten Alkohols unter Umrihren vermischt, so bildet sich ein Niede:schlag ; nach 15-24 stiindi- gem Stehenlassen wird die Fliissigkeit abgegossen und der zuriickbleibende Niederschlag nochmals mit absolutem Alcohol versetzt und dieselbe Operation wird so lange wiederholt, bis der zugesetzte Alcohol vollkom- men farblos erscheint. Nach Abpressen zwischen Filtrirpapier wurde der Niederschlag bei 50-609 C. getrocknet: es resultirt eine gelblich weisse leicht zerreibliche Masse. Eine Einspritzung von 0,0005 g. dieser Substanz bei tuberculésen Meerschweinchen ruft eine deutliche Steigerung der K6rpertemperatur hervor. Es erwies sich schwacher als das Koch’sche Praeparat, wesshalb ich nun Ammonsulfat anwandte. IV. Ammonsultatfallung des Tuberculins. Mein Tuberculinpraeparat wird mittelst Ammonsulfatfallung gewonnen. Sattigt man fliissiges Rohtuberculin mit Ammonsulfat, so scheidet sich eine gelbbraunliche zihe Masse ab, die nach 24 Stunden gesammelt und einmal mit gesattigter Ammonsulfatlésung ausgewaschen, dann zwischen Filtrirpapier gepresst, uud in etwas destillirten chloroformhaltigen Wasser gelést, hierauf so lange gegen str6mendes Wasser dialysirt wird, bis die Lésung keine Reaction auf Schwefelsiure mehr zeigt. Die Fliissigkeit wird dann filtrirt, das Filtrat bei 50-60° C. eingedampft und schliesslich mit cinem Ueberschuss von absolutem Alcohol unter Umriihren versetzt. Nach 24-48 Stunden wird der voluminése gelblich weisse Niederschlag abfiltrirt, 2-3 Mal mit absolutem Alkohol ausgewaschen, zwischen Filtrir- papier gepresst und bei 50-69° C. getrocknet. Das dabei gewonnene grauweisse Pulver ist leicht loslich und enthalt nur 1.5446% Asche, wihrend Koch’sche Praeparate an 20% Asche enthalten. Die wisserige Lésung 124 N. Nitta: dieser Substauz reagirt neutral und ist von braunlicher Farbe. Dieselbe coagulirt nicht beim Kochen und wird durch Salpetersdure sowie durch Ferrocyankalium und Essigsaure gefallt ; der dabei gebildete Niederschlag lést sich beim Erwarmen und scheidet sich beim Abkiihlen wieder ab. Dieselbe wird auch durch Phosphorwolframsaure, Pikrinsaure, Gerbsiure, Sublimat und Ammonsulfat gefallt: Biuret, Millon’sche uud Xanthoprotein- reactionen fallen positiv aus. Dass mein Praeparat frei von peptonartigen Substanzen ist, geht daraus hervor, dass dasselbe vollstandig durch Ammonsulfat gefallt wird und das Filtrat von der Ammonsulfatfallung keine Peptonreactionen gibt. Diese peptonfreie Substanz ist daher als Tuberculinalbumose zu_be- zeichnen. Die wasserige Loésung meiner Tuberculinalbumose gibt bei Sattigung mit Chlornatrium sehr schwachen, beim weiteren Zusatz von etwas Essigsiure aber starken Niederschlag, was andeutet, dass meine Tuber- culinalbumose hauptsichlich den Character einer Deuteroalbumose von Kiithne tragt; sie enthalt aber noch Spuren von Protoalbumose. Die Thatsache aber, dass eine wasserige Losung meines Praeparates mit verdiinnter Essigsaiure einen schwachen im Ueberschuss derselben léslichen Niederschlag erzeugt, deutet darauf hin, dass sie eine geringe Menge einer Albumose enthalt, welche der Atmidalbumose Neumeister’s analog ist. V. Injectionsversuche. a) Mit Meerschweinchen. Die in destillirtem Wasser geléste Tuberculinalbumose wurde in verschiedenen Dosen tuberculésen Meerschweinchen (K6rpergewicht: ca 400-5co g.) subcutan eingespritzt. Die dabei gewonnenen Ergebnisse sind aus folgender Tabelle zu ersehen: Ueber das wirksame Princip des Tuberculinum Kochii., 125 S & vb Korpertemperatur, Cels,° * =a Mee Bo, oo Gas bh Ze - S a = Nach Einspritzung (Stunden). eh 5 od 5= 3 bo se oe PB, o.5 el 5 | 2 = =) 27 ha as 2 — 38,4 38,8 39,25 | 40,0 40,0 39,8 39,6 rig: 37:35 | 37,95 | 39:25 | 39:75 | 49,05 | 40,05 | 39,85 | — 2.8 38,2 38,6 38,8 39,15 | 39,8 40,2 39:7 = 2.0 389 | 389 | 39% | 39:7 | 395 | 394 | 39.3 | 39:0 1.2 39,4 | 40,2 | 40,2 | 39.9 | 39:2 | 39,0 | 39.1 | 38,9 1.6 — | 391 | 39,3 | 395 | 40,1 | 40,1 | 40,0 | 39,6 1.0 a 39,0 39,2 39:4 39.1 39,1 39:4 39,2 0.4 Bei Betrachtung der obigen Versuche erkennt man, dass die Tuber- culinalbumose in Dosen von 0,00001 (—o0,000005) g. bei tuberculésen Meerschweinchen die echte Fieberreaction zu erzeugen im Stande ist. Beim Vergleich der Wirkung des Roktuberculins und des Reintuberculins von Koch mit meiner Tuberculinalbumose ergab sich folgender Unter- schied : IPOnEM Be CCU NAC Ne KOCH sie. viecccsccec cscs eceneeee 0,001 ccm. PSCIMEWHEHCUINT NACH IGOCN © Joccl ccc ccc cees sce ences 0,00005 ¢ PSE ECT Micey EUIINIOSE caus dec oe teaaycsdesvseesseservee 0,O000I ¢. Die Versuche beziiglich ihrer tétlichen Wirkung auf tuberculése Meer- schweinchen (Kérpergewicht: ca. 400-500 g.) hatten folgendes Ergeb- niss: 126 N. Nitta: y : 5 bb K6rpertemperatur, Cels.° a 2 8 : irpertemperatuy e 3 eee ee sp > 28 a} 5 2 Nach Einspritzung, (Stunden.,) Bemerkungen, wn Q u iS 4s) Se] twa Sis Snes | cae: ieee 2 sa | s I Todt. 2 ” 2 Todt in der 2 folgenden Nacht. 4 Todt. Todt in der 5 folgenden Nacht. 6 ” Lebend. Aus obigen Tabellen ersieht man, dass die minimale Lethaldose der Tuberculinalbumose fiir mittelgrosse Meerschweinchen 28-48 Tage nach der Impfung mit Tuberkelbacillen 1 Milligr. ist. Bei den Tieren, welche dieser Dose erlagen, finden sich bei den Eingeweiden, insbesondere an der Oberflaiche der Milz und Leber, zahlreiche haemorrhagische Flecke von Mohnsamen- bis Pfenniggrésse. Diese Haemorrhagie wurde auch von mir bei der Rohtuberculininjection beobachtet: nach R. Koch ist dieser Befund ein charakteristisches Merkmal der Tuberculinwirkung. Nach Koch erfordert die Tétung der Tiere von seinem Reintuberculin 5-10 Milligr: somit ist Giftwirkung meiner Tuberculinalbumose 5-10 fach so stark als die der Koch’schen. Zum Vergleich habe ich auch einige Tierversuche mit den von mir aus Witte’schem Pepton hergestellten Albumosen ausgefuhrt. Die Injec- tion von o.1 g. dieses Praeparats hatte nur einige Zehntel Grade Tem- peratursteigerung im Gefolge und schidigte das Wohlbefinden der Tiere nicht im Geringsten, wahrend von meiner Tuberculinalbumose schon 0,001 g. tdtlich auf diese Tiere wirkt und 0,oooo1 g. schon eine Tem- peratursteigerung herbeiftihrt. ‘asseY JOUIOIS[OP] UL oyINsIOA “I pun £1 z a a *Sunznory-us0yysoys uv ayonsi9A ‘ZI-I x N 5S ¢ = ; as 3 ; = op seen ET) : “ “ “ z VI e ; e ‘ FOE!) 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Man erkennt, dass sammtliche Tiere, welche auf Rohtuber- culinimpfung reagirt haben, eine starke Fieberreaction auf Einspritzung meiner Tuberculinalbumose zeigen. Im Anschluss an diese Tierversuche habe ich noch mit anderen aus dem Rohtuberculin hergestellten K6rpern und zwar mit dem Aether- extract, welches aus dem mit verdiinnter Schwefelsiure versetzten Rohtu- berculin gewonnen wurde, sowie mit dem direkt hergestellten Aether- extract an tuberculésen Meerschweinchen (mittelgross) Versuche gemacht: dabei liess sich aber nirgends eine tuberculinahnliche Wirkung beobachten. Ferner zeigten tuberculdse Meerschweinchen (mittelgross) nach Einspritz- ung von Bernsteinsaéure (0,01I-0,05 g.) oder von bernsteinsaurem Natron (0,01-0,1 g.) gar keine Steigerung der K6rpertemperatur, was die Behaup- tung Viquerat’s, dass diese Saure das active Princip im Rohtuberculin sei, widerlegt. VI. Chemisches Verhalten meiner Tuberculinaibumose. Eine 2% ige Lésung der Tuberculinalbumose zeigt folgendes Ver- halten: I. Alcohol 96% : Mit gleichem Volum fallen zarte, weisse Flocken aus. 2. Salpetersiure in der Kialte: Niederschlag, der sich in der Warme rasch lést, in der Kalte aber wieder auftritt. 3. Gleiches Volumen conzentr. Kochsalzlésung zu der mit Essigsiure angesduerten Loésung gesetzt: miassiger Niederschlag, beim Er- hitzen nicht léslich. 4. Siattigung der neutralen Lésung mit Kochsalz: Spur Triibung. Verdiinnte Kupfersulfatlésung : starke Fallung. ae Essigsiiure-Ferrocyankalium: starke Fiallung, léslich beim Erwar- men. I 30 N. Nitta: 7. Pikrinsdure: starke Fallung. 8. Metaphosphorsaure: Fallung, im Ueberschuss wieder léslich. 9. Trichloressigsaure: starke Fallung, in der Hitze léslich, in der Kalte wiederkehrend. 10. Jodquecksilberkalium: erst in saurer Lésung starke Fallung, welche sich im Ueberschuss von Salzsaure nicht lost. 11. Gerbsaure: starke Fallung, die sich in der Hitze nicht lést, © 12. Millon’sches Reagens: weisse Fallung, beim Kochen Rothfarbung. 13. Xanthoproteinprobe: starke Fallung, welche sich im Ueberschuss von Salpetersaure lést, beim Erhitzen Gelbfarbung. 14. Adamkiewiez’sche Reaction: Violettfarbung. 1s. Molisch’sche Zuckerprobe: schwach _ positiv, beim Erhitzen schwache Violettfarbung. 16. Biuretprobe mit Kupfersulfat : positiv. 17. Biuretprobe mit Nickelsulfat: beim Erwarmen schwache Gelb- farbung. 18. Kochen mit Alkali- und Bleiacetat : Schwarzfarbung. Nach diesen Reactionen entspricht meine Tuberculinalbumose am nachsten der Deuteroalbumose Kiihne’s, ferner in einem gewissen Grade der secundaren Albumose A von Pick; jedoch mit dem Unterschied, dass meine Albumose bei den oben unter 3 und 6 erwaihnten Reactionen einen starken Niederschlag gibt, wahrend die secundire Albumose A Pick’s hierbei nur Spur Triibung gibt; die secundire Albumose B. und C. gibt hier gar keine Triibung. VII. Hitzebestandigkeit der Tuberculinalbumose. Kine 19§ ige wiasserige Lésung der Tuberculinalbumose wurde in kochendem Wasser je 10 Minuten, 30 Minuten und eine Stunde lang erhitzt, dann tuberculédsen Meerschweinchen subcutan eingespritzt ; fol- gende Tabelle zeigt die dabei gewonnenen Ergebnisse : Ueber das wirksame Princip des Tuberculinum Kochii. 131 K6rpertemperatur, Cels.° OD = b ae bho ve a Se 3S | Erhitzungs Bee Zo a2 eee S = Nach Einspritzung. (Stunden.) mig 2g auer. 2 a 5 Os eee o oh = 5 1 oD) = Co te 1 | Zehn Minuten. 3 38,3 17 2 253 3 28 Dreissige * 2 c - 4 Minuten, 2,45 5 ” , 3 : 1,75 6 Ein Stunde, 3 3,0 i ” M 2,6 5 Controlldsung, 3 3 2,6 9 ” 3 40,7 3 5 333 Es unterliegt also kaum einem Zweifel, dass die Tuberculinalbumose gegen Hitze sehr widerstandsfahig ist. Eine 1 stiindige Erhitzung bei 100° C. ist nicht im Stande, die Wirksamkeit der Tuberculinalbumose im Geringsten zu schiadigen. VIII. Verhalten der Tuberculinalbumose gegen Pepsin und Trypsin. Je 0,1 g. Tuberculinalbumose wurde einerseits in 10 ccm. 0,2% iger Salzsiure und andererseits im gleichen Volum 0,2% iger Sodalésung (Na,CO,) gelést und dort etwas Pepsin, hier aber ebensoviel Trypsin zugesctzt. Bei Gegenwart von etwas Chloroform wurden beide Lésungen wohlverschlossen 2 Tage lang im Brutofen bei 37-38° gelassen. Hierauf wurden sie im Dampftopf 10 Minuten lang erhitzt und tuberculésen Meer- schweinchen subcutan eingespritzt ; gleichzeitig wurden Controllésungen (Tuberculinalbumoselésungen in 0,2% iger Salzsaure und in 0,2% iger Sodalésung ohne Pepsin resp. Trypsin) gepriift: Die Ergebnisse sind in folgenden Tabellen zusammengestellt : I 52 N. Nitta: teks a * x & 0 | K6rpertemperatur. Cels.° 2, ae) .Po o . . Two iad 9 £ ie 8°S | Tubercul ae BO 23 uberculin- BE Nach Ejinspritzung, (Stunden. 2 a ls 8 =e albumose. go Sia £5 oe © tp 5S tae farto Z is Ae S3 & Mit Pepsin. 2 1,0 3 oe 4 | Ohne Pepsin, 1,9 5 = 0,0002 Syria | 2,6 6 Mit Trypsin, } 0,001 38,0 0,6 7 0,8 8 0,8 9 | Ohne Trypsin. | 0,oo1 37,4 22, Io ~ 0,0002 | 37,9 17 Die Tiere verhalten sich also bei Behandlung mit verdauter Tuber- culinalbumose indifferent; Pepsin und Trypsin hatten die Tuberculin- albumose verindert. IX. Sind gewisse labile Atomgruppen die Ursache der Giftwirkung? Die schon erwahnte Thatsache, dass die Tuberculinalbumose bei einstiindigem Erwarmen nicht im Geringsten an Wirksamkeit einbiisst, liess es von vorneherein wenig wahrscheinlich erscheinen, dass die Wirk- samkeit auf besonders labilen Atomgruppen beruhe. Nichtsdestoweniger wurden einige Versuche angestellt mit Kérpern, welche sehr leicht in labile Amido- und Aldehyd- oder Keton- gruppen eingreifen. Je 5 ccm. 20% ige wisserige Lésung des Rohtuberculins wurde mit 25 ccm. der folgenden Lésungen vermischt (in jedem Falle zwei Kélbchen). 1. Controllésung. 2. 1% Natriumnitritlésung: unmittelbar vor Gebrauch mit einigen Tropfen verdiinnter Essigsaure versetzt. Ueber das wirksame Princip des Tuberculinum Kochii. 133 3. 5% Formaldehydlésung. 4. 1% Hydroxylaminchloridlésung: unmittelbar vor Gebrauch mit Natriumcarbonat neutralisirt, (entspr. 0,47 freiem Hydroxylamin). Die Mischungen wurden mit etwas Chloroform versetzt und 24 Stunden bei Zimmertemperatur stehen gelassen, dann mit absolutem Alcohol ausgefallt, der Niederschlag wiederholt mit absolutem Alcohol ausge- waschen und nach Pressen zwischen Filtrirpapier in wasserigen Lésungen tuberculésen Meerschweinchen subcutan eingespritzt. Die dabei gemach- ten Beobachtungen zeigt die folgende Tabelle : 7 3 ; Ko6rpertemperatur, Cels.° g. oi) 5 Oh oF 5°O 3 £5 é & Reagentien, 8 uf Vor Nach Einspritzung, (Stunden.) 3 ais Sg {Einspritz- 22 A as ung Ae 3 : th 2h 3h 4h 5h 6h 7h 2 1 { Controllésung. 38,6 — | 39,0 | 40,8] 40,6] 40,4 | 40,0 | 37,0 2,2 2 | Natriumnitrit. 38.4 | — | 388 | 39,2 | 39,9] 39,7] 39,7] 396] 15 3 ” 37,8 5 SE, 38,65 38,8 40,0 39,7 40,0 39,6 2,2 4 ” 38,5 | -- | 390 | 39,3 | 39,7] 39.4] 39,0] 39.0] 12 5 | Formaldehyd. 3759 — | 37:9 | 40,0] 40,3 | 40,0 | 40,0 | 40,0 2,5 6 ” 38,2 | — | 381 | 39.4] 39,6] 39.5] 39.2] 392] w4 ie ” 38,5 | — | 382 | 39.4] 40,2] 40,2} 30,81 39,5] 17 Hydroxyl- : 8 BErechlodd. 379 | — | 380 | 38,3] 30,5 | 39:7] 397 | 40,2 253 9 » 38,0 | — | 38,15] 39,2 | 40,0] 40,5] 40,3] 40,2] 2,5 10 » 38,6 — | 39,0 | 39,7] 40,1 | 40,4] 40,1 | 39,7 1,8 Hieraus ersieht man, dass oben erwahnte Reagentien keinerlei schadi- genden Einfluss auf das Tuberculin hatten, dessen Natur demnach wohl verschieden ist von derjenigen der gew6dhnlichen Enzyme; denn diese werden durch 5% Formaldehyd nach 24 Stunden stehen leicht unwirksam. 134 N. Nitta: Schlussfolgerungen. Die Resultate meiner Untersuchungen sind wie folgt : 1. Die Ammonsulfatfallung des Tuberculins dussert bei Meerschwein- chen und Rindern sich qualitativ wie das Koch’sche Reintuberculin ; es reichen jedcch von meinen Praeparat weit geringere Mengen (4) hin, dieselbe Reaction zu erzeugen. 2. Die Ammonsulfatfallung besteht wesentlich aus einer Deuteroalbu- mose nebst Spuren Prot- und Amidalbumose (Tuberculinalbumose Nitta’s). 3. Unter der Einwirkung von Pepsin und Trypsin verliert meine Tuberculinalbumose ihre specifische Wirkung. 4. Nach einstiindigem Erhitzen auf 100° C. behalt die wasserige Lésung der Tuberculinalbumose ihre specifische Wirkung. 5. Die specifische Wirkung des Tuberculins verandert sich nicht im Geringsten unter der Einwirkung von Natriumnitrit (19%), Formal- dehyd (5%) und Hydroxylamin (0,479%). 6. Bei Tierversuchen zeigt die Ammonsulfatfallung von Witte’schem Pepton nicht die gleiche Wirkung wie das Tuberculin resp. die Tuberculinalbumose. 7. Das wirksame Princip des Tuberculins ist eine Albumose (Deutero- albumose). 8. Die Wirkung der Tuberculinalbumose, des wirksamen Princips des ,,luberculins,” ist ganz specifisch, und durch gewohnliche Albu- mosen nicht herbeizufiihren. Die Kiihne’sche, Hunter’sche und Helmann’sche Ansicht beziiglich des wirksamen Princips des Tuber- culins ist unrichtig. Zum Schlusse halte ich es mir eine angenehme Pflicht, Herrn Prof. Dr. O. Loew und Herrn Prof. Dr. Y. Kozai fiir ihre giitige Unterstiitzung meinen ergebensten Dank auszusprechen. to 6. SS DY. 14. se Ueber das wirksame Princip des Tuberculinum Kochii. Literatur. Buchner, H., Tuberculinreaction durch Proteine nicht specifischer Bacterien, Miinch. Med. Wochenschr. 1891. No. 49. Dixon u. Zuill, Reaction of the Amido-group upon the Wasting Animal Economy. Philadelphia (The Amer. Med. Press Comp.) 1891. Helman, C., Des Propriétés de la tuberculine provenant de Bacilles tuberculeux cultivés sur pommes de terre, Archiv des Sciences biologiques publ. p. l'Institut impér. de méd. expér. a St. Péters- Nouro. f..1... No. 1 u..2. Hunter, W., On the Nature, Action and Therapeutic Value of the Active Principles of Tuberculin, Brit. Med. Journ. 1891 July. 25. Hutyra, F., Tuberculinversuche bei Rindern: Nachtrag, Zeit- schrift f. Thiermedicin. Bd. IV. Heft. 1. Klemperer, G., Die Beziehungen verschiedener Bakteriengifte zur Immunisirung und Heilung, Zeitschrift f. klin. Medicin, Bd. XX. Pief tu. 2. Koch, R., Weitere Mittheilung iiber ein Heilmittel gegen Tuber- kulose, Deutsche medicin. Wochenschrift, 1890, No. 46 a. Derselbe, Fortsetzung der Mittheilungen iiber cin Heilmittel gegen Tuberkulose. Ebendas. 1891. No. 3. Derselbe, Mittheilungen tiber das Tuberkulin. Ebendas. 1891. No. 43. Kihne, W., Weitere Untersuchungen iiber die Proteine des Tuber- culins, Zeitschrift f. Biologie, Bd. XXX. (N. F. Bd. XII.) Liebreich, O., Ueber Lupusheilung durch Cantharidin und iiber Tuberkulose, Berl. klin. Wochenschrift, 1895, No. 14, 15. Matthes, M., Ueber die Wirkung einiger subcutan einverleibten Albumosen auf den tuberculés inficirten Organismus, Deutsche Archiv f. klin. Medicin, Bd. 54, Heft. 1. v. Mosetig, Teucrin, Wiener med. Presse, 1893, No. 6. Neumeister, R., Lehrbuch der physiolog. Chemie. 1897. Petri u. Maassen, Beitrige zur Biologie der krankheitserregenden 136 N. Nitta: Ueber das wirksame Princip des Tuberculinum Kochii. Bakterien, insbesondere iiber die Bildung von Schwefelwasserstoff durch dieselben unter vornehmlicher Berticksichtigung des Schwei- nerothlaufs, Arbeiten aus dem kaiserl. Gesundheitsamte, Bd. VIII. Pick, E., Untersuchungen wtber die Proteinstoffe, Zeitschrift. f. physiolog. Chemie, Bd. XXIV, Heft. 3. Roemer, G., Tuberculinreaktion durch Bacterienextrakte, Wiener klin. Wochenschrift. 1891, No. 45. Ruppel, G., Zur Chemie der Tuberkelbacillen; Erste Mittheilung, Zeitschrift f. physiolog Chemie, Bd. XX VI. Heft. 2 u. 3. Spiegler, E., Ueber Localreaction in Folge hypodermatischer Einverleibung chemischer Verbindungen, Centralblatt f. klin. Medicin, 1893, No. 36. Viquerat, Beitrag zur Tuberculinfrage, Centralblatt f. Bakt., Bd. XXXVI. “No. 10: ee Ueber Ernahrungsverhaltnisse beim Bacillus prodigiosus, VON O. Loew und Y,. Kozai. Da aus mehreren Beobachtungen die Bildung eines bacteriolytischen Enzyms beim Aac. prodigiosus wahrscheinlich wurde, stellten wir einige Versuche an, welche Aufklarung dariiber geben sollten, inwieweit die Bildung eines solchen Enzyms mit Ernahrungeverhiltnissen bei diesem Microben zusammenhinet. /eriz beobachtete, dass Zucker mit Ammoniak- salzen die Bildung eines proteolytischen Enzyms verhindert, dagegen Glycerin mit Ammoniaksalzen dieselbe begiinstigt. Ob dieses proteoly- tische Enzym zugleich ein bacteriolytisches ist oder neben dem ersteren ein bacteriolytisches vorhanden ist, wurde nicht untersucht. Die Existenz eines bacteriolytischen Enzyms in Culturen das 2. prodigiosus schien aus der Beobachtung von Freudenreich? hervorzugehen, dass diese Culturen sehr entwicklungshemmend auf einige andere Bacterienarten wirken. Bei unserer ersten Versuchsreihe verwendeten wir folgende Loesungen : Pepton 1% mit Glycerin 0.1%. ) ” ” ») en: he », Mit essigsaurem Natron 0.2% und Asparagin 0.2%. _ . Pepton 0.2% mit essigsaurem Natron 1%. Asparagin 0.2% mit Glycose 1%. Harnstoff 0.29§ mit Glycose 1%. Natriumnitrat 0.2% mit Glycose 1%. ey ARR wd Bouillon. Die zugesetzten Mineralsalze bestanden hier aus : 1 Arch Hyg. Bd. 14; S. 16 und 30. 2 Jahresber. f. Bakt, 1889, S. 531. 138 0. Loew und Y. Kozai; Sécundarem Kaliumphosphat. 2... ...48:.. 2.0. o.2 meee Natriuiweulfat.. 2.32. 2. sete ecg ee Seo. rnc ee ON Ge Magnesiumsultat .-saneigeee yan. ee 2 ne ee 0.01%. Die Proben wurden zuerst zwolf Tage bei 10-15°, dann noch funf Tage im Brutkasten gehalten. Es ergab sich dann folgendes Resultat : Loesung 1 und 2: Viel Bacteriensediment, und wenig Farbstoff. 3: Nach anfanglich reichlicher und rot gefarbter Vegetation fast vollige Wiederloesung. 4. 5 und 6: Geringe Entwicklung. 7: Gar keine Entwicklung. 8: Miassige Entwicklung, rotes Sediment,! welches selbst nach acht weiteren Wochen nicht wieder gelost war. Die Combination von Pepton mit essigsaurem Natron und Asparagin hatte sich der Bildung von Farbstoff und bacteriolytischem Enzym am giinstigsten erwiesen. Wo das stickstoffhaltige Material gegeniiber dem stickstofffreien vermindert war, fand nur geringe Entwicklung statt, bei Natriumnitrat als Stickstoffquelle gar keine. Bei unsrer zweiten Versuchsreihe ersetzten wir das schwefelsaure Natron durch Chlornatrium, machten ferner die Loesungen schwach alkalisch und machten geringe Zusatze von Stoffen, welche bei héheren Concentrationen giftig wirken, um zu beobachten, ob eine giinstige Einwirkung auf Wachsthum und Enzymbildung stattfinden wiirde.? Unsre Controlloesung hatte die folgende Zusammensetzung : Pepton °..: 0s eens eet Rate eer 0.5%. Glycerin 2... s0cjiee ets Oe eee Onli Dikaliumphosphat*:....°-A'00 2%. oa eee ee oe Natriumbicarbonath. 1.05. ee ee eee ROM se Natriumchlorid. .s-c..<.0 bak eee a ee 0.2 3 Magnesiumsulfat ccc peers eee. oo ee 0.01%. | Nach Avwn/ze sind Magnesia sowohl wie Schwefelsdure wesentlich fiir die Farbstofiproduction. Jedenfalls existiren aber hier auch noch manche andere Finfliisse, auch solche, welche der Production entgegen_wirken. 2 Nach //iipfe's biologischem Grundgesctz wircken Gifte bei sehr hoher Verdiinnung als Reizmittel, Ueber Ernahrungsverhaltnisse beim Bacillus prodigious, 139 In Loesung 2 war das Chlornatrium durch die aequivalente Menge (0.25%) Natriumsulfat, in 3 durch die aequivalente Menge Natriumnitrat (0.30%) ersetzt. In 4. war der Normalloesung noch 0.01% Jodkalium, in 5. ebensoviel Fluornatrium, in 6. ebensoviel Ferrocyankalium zugesetzt worden. Die dreimal sterilisirten Loesungen wurden am 16. Januar inficirt und bei 6-15° C. stehen gelassen. Am 27. Januar war der Stand folgender : 1. (Control): Keine Haut, nur Triibung uud ein Ring am Rande. 2. (Na,SO,): Roter Ring, Spur Haut, Triibung. 3. (NaNO,): Loesung klar, keine Haut, nur schwacher weisser Ring. 4. (NaJ): Triibung, schwache rote Haut. 5. (NaF): 2 rs ie re 6. (Kfcy): Triibung, stark entwickelte rote Haut. Am 3. Februar wurde bemerkt, dass die Farbung am intensivsten in 5. war, diese verblasste aber spater wieder; die Vegetation war am iippigsten in 6. Am 18, Februar war der Stand folgender : Bei 3. Triibung, geringer weisser Bodensatz , tunds5: Méassiger Bodensatz, kaum gefarbt. 2und 4: Etwaebenso starkes Wachsthum als bei 1 und 5, aber mehr Farbe. 6: Der Bodensatz betrug hier, dem Volumen nach abgeschitzt, mindestens des werfache der in 1. gebildeten Masse; es war also eine Retswirkung des Ferrocyankaliums auf die Wachs- thumsintensitaet unverkennbar. Wahrend in der ersten Versuchsreihe das Natriumnitrat als untaugliche Stickstoffquelle erschien, hat es sich in dieser als sehr hemmend selbst bei Anwesenheit von Pepton erwiesen.'| Fluornatrium und Jodkalium haben 4 Da die Hemmung von Anfang an vorhanden war, so kann sie nicht etwa die Folge von erst gebildetem Nitrit sein, Die Ursache ist nicht leicht anzugeben, es mag aber darauf hingewiesen werden, dass Nitrate auch hemmend auf die Entwicklung der Legumisnosenbacterien wirken (Jfarcha/, Compt, rend. 133 p. 1032) sowie auf die Kohlensiureassimilation der Meeresalgen (4réer, Botan, Centralbl, 1902 p, 120) und schliesslich auch auf die katalytische Wirkung der Katalase. 140 in der angewandten Verdtinnung das Wachsthum nicht geférdert, wenig- 0. Loew und Y. Kozai: stens nicht in direct erkennbarem Maase. Die auffallende Wirkung des Ferrocyankaliums veranlasste uns zu einem weiteren Versuch, in welchem diese Wirkung auf den B. prodigiosus mit der auf andere Microbenarten verglichen wurde. diente hier Bouillon, je 8 cc. in einer Eprouvette. Als Nahrioesung Nach drei Tagen bei 35° war das Resultat folgendes : Microbenart. B, prodigiosus, Bouillon, Ohne Zusatz, Kein Bodensatz, schwacher roter Ring. B. peyoyaneus, B, mesenter, ruber, Dicke Haut, Starke Haut. B. megatherium, Haut, B, Zenkeri, Triibung u, Flocken. B, cyanogenus, B. capsulatus, B, acidilactici (ppc. I5. subtilis, B, typhi mur, Ks hat sich also eine stimulirende Wirkung nur beim B. prodigiosus erkennen lassen, in den andern gepriiften Fallen ergab sich meistens einc deutliche Schadigung. beim B, prodigiosus es sich um eine Spaltung des Ferrocyankaliums handelt, wobei Haut und Bodensatz. Dunkle Varbung nahe der Oberfliche. Triibung, starker Rand, Starker Bodensatz. Dicke Haut, Tribung. einerseits die schadigende Cyanwasserstoffsaure sofort Dieses Resultat veranlasst uns zur Annahme, dass Mit 0.019% Ferrocyankalium, | Starker Bodensatz, dicker roter Ring. Schwache Haut, Schwache Haut. Nur Trtibung und Ring. Trtibung und Flocken. Geringe Ilaut und Bodensatz, schwiichere Fiirbung. Triibung, schwacher Rand, Schwacher Bodensatz. Rit s- Dunne Haut. Tribung, AMS lee Ueber Ernahrungsverhiltnisse beim Bacillus prodigious. 141 weiter zersetzt wird, andrerseits das in den Zellen freiwerdende Eisen in einer lockeren salzartigen Verbindung eine fordernde Wirkung ausiibt. Fassen wir unsre Beobachtungen am 2. prodigtosus kurz zusammen, so ergibt sich Folgendes : I. Eine fiir die Production von Farbstoff und bacteriolytischem Enzym giinstige Nahrstoffcombination besteht aus Pepton 1%, essigsaurem Natron 0.2% und Asparagin 0.2%. Eine betrichtliche Vermehrung stickstofffreien Materials gegeniiber stickstoffhaltigem tibt auf die Entwicklung einen ungiinstigen Effect aus. Natriumnitrat ist nicht nur unfahig, als Stickstoffquelle zu dienen, sondern hemmt sogar die Entwicklung bei Gegenwart von Pepton. Jodkalium und Fluornatrium in einer Verdiinnung von 0.1 p.m. iiben keine deutliche Reizwirkung aus. Ein Ferrocyankaliumzusatz von 0.1 p.m. befordert die Entwicklung, was bei andern Microben nicht zutrifft. z . eas aA Ueber die Vertheilung des Kalks im thierischen Organismus., VON M. Toyonaga. Die Wichtigkeit des Kalks fiir alle thierischen Organismen ist seit lange anerkannt. Ungefahr drei Viertel simmtlicher Mineralstoffe der héheren Thiere besteht aus Tricalciumphosphat, da dieses Salz die Hauptmasse der Knochen und der Zahne ausmacht. Aber abgesehen von diesem mehr in die Augen fallenden Vorkommen finden sich noch Kalk- verbindungen in sammtlichen Organen vor, ja hoéchst walrscheinlich existiert auch bei den niedersten thierischen Organismen keine Zelle, die frei von Kalkverbindungen wire. Auch die grosse Wichtigkeit fiir das Blut ist in neuerer Zeit anerkannt worden. Blut verliert seine Gerinn- barkeit, wenn durch Zusatz von etwas Natriumfluorid oder Natriumoxalat der Kalkgehalt des Blutes in die unléslichen Formen Calciumfluorid oder Kalkoxalat iibergefiihrt wird. Auch mag darauf hingewiesen werden, dass in kalksalzfreier Lésung auch das Casein der Milch auf Labzusatz nicht gerinnt, wohl aber wird dabei das Casein so veriindert, dass nun auf Zusatz von Kalksalzen ein Niederschlag yon den Eigenschaften des frischen Kises entsteht. Cavazzani hat es ferner wahrscheinlich gemacht, dass Kalksalze auch fiir die Gerinnung des Muskelplasmas von Bedeutung sind. Im Harne findet sich Kalk regelmiassig gelést, fernecr ausnahmsweise in Form von Concretionen. Die Anwesenheit von Kalk in den Nahrungsmitteln ist bei dem Bedutrfniss von. Thier und Mensch fiir Kalk von grésster Wichtigkeit. Junge Thiere leiden bei kalkarmer Nahrung an Rachititis ahnlichen Veranderungen, aber auch ausgewachsene Thiere leiden in Folge eines solchen Mangels, da die Kalkausscheidung durch die Niere regelmiassig fortdauert. Bunge hat deshalb auf die Gefahr hingewiesen, wenn man die 144 M. Toyonaga: an Kohlehydrat reichen Vegetabilien der menschlichen Nahrung durch blosen Rohrzucker ersetzt. Erstere enthalten verschiedene mineralische Nahrstoffe wie Phosphate, Kalk, Magnesia und Eisen- Salze, wiahrend dem Rohrzucker des Handels, der reinen Saccharose, alle diese Stoffe mangeln, besonders aber fallt der Mangel von Kalk und Eisen ins Gewicht. Bunge glaubt, dass Animie und Zahncaries auf den mit Zuckergenuss ver- bundenen Kalk- und Eisenmangel zuriickzufiihren sind und er schlagt desshalb vor, das Bediirfniss der Kinder nach Siissigkeiten, statt mit blosem Zuckerpreparaten, mit stissen Friichten, frisch oder gekocht, zu befriedigen. Auch fiir unsere Hausthiere ist es von sehr grosser Bedeutung, dass bei Zusammensetzung einer Futterration der Kalk dieselbe Beriick- sicktigung als andre Niahrbestandtheile findet. Vielfach ist Knochen- briichigkeit des Rindsviehs auf eine ungentigende Ernahrung mit Kalk zuruckzufiihren. Die Thiere werden unter solchen Umstanden matt und magern ab und bei den geringsten Veranlassungen treten Knochenbriiche ein. In manchen Gegenden mit kalkarmen Boden zeigt sich in Folge des kalkarmen Grases die Knochenbrichigkeit sehr haufig beim Rind, welches auf diese Nahrung angewiesen ist. Katsuyama hat gefunden, dass Kaninchen beim Hungern anfangs etwas weniger Kalk ausscheiden, die Menge steigt aber vom vierzehnten Hungertage an langsam bis zum Tode. Die Magnesiawerthe dagegen zeigen ein continuirliches Sinken. Gerhart und Schlesinger fanden, dass die Ausscheidung des Kalks im Harne beim Gesunden und beim Diabetiker der Ammoniakauscheidung parallel geht und wie diese durch Alkalizufuhr herabdriickbar ist. Unter abnormaler Saurebildung im Thier nimmt auch der Kalkgehalt des Harnes abnorm zu. Eine Vermehrung des Kalkgehaltes des Harnes oder der Faeces findet nach Babeau auch in der Entwickelungsperiode der Rachitis statt. Geléste Kalkverbindungen sind ferner nach //ad/zburton fir die Herzthatig- keit von grosser Bedeutung, was von Fagues Loed bestatigt wurde. Es ist daher von grésstem Nachtheil fir die regelmassige Herzthatigkeit, wenn der Kalk des Blutserums durch Fluornatrium oder oxalsaures Natron? in 4 Zeitschrift f, physiol, Chemie 26, S. 542. 2 Fluornatrium hat tibrigens auch noch andre Wirkungen, die nicht auf der Kalkfallung beruhen, ? ! Ueber die Vertheilung des Kalks im thierischen Organismus. 145 unlésliche Verbindungen iibergefithrt wird. In neuester Zeit hat /yzeden- thal? es sehr wahrschcinlich gemacht, dass die Giftwirkung der Seifen (oelsaures Natron) bei Injection in das Blut auf der kalkfallenden Wirkung derselben beruht. Er sagt richtig: ,,wie hatte man vermuthen k6nnen, dass eine Substanz, die in grossen Dosen verfiittert werden kann fast wie ein Nahrungsmittel, die in kleinen Mengen einen normalen Bestandtheil der Gewebe und des Blutes bildet, in die Blutbahn cingefiihrt schon in Dosen von 0,1 g. pro Kilo Thier den Tod im Augenblick der Einfiihrung herbeifuhren kénne. J/uzk hat gezeigt, dass es nicht etwa das aus den Seifen freiwerdende Natron ist, welches die Giftwirkung bedingt. Dieses wiirde ja auch bei dem Kohlensiure gehalt der Blutes sofort in Carbonat iibergehen. Obwohl nun jede thierische Zelle Kalksalze benéthigt, so existirte doch bis in die neuere Zeit herein keine befriedigende Theorie der Funk- tionen des Kalks fiir die thierischen Zellen. Auch Pflanzen mit Ausnahme der niedersten Formen bediirfen des Kalks. Was nun diese Funktion des Kalks in den Pflanzen betrifft, so hat O. Zoew aus der Giftwirkung des neutralen oxalsauren Kalis, welche er unter dem Mikroskop verfolgte, eeschlossen, dass sowohl die Chlorophyllkérper als auch der Zellkern aus Kalkverbindemgen von Nucleoproteiden aufgebaut sind. Die Funktion der Magnesia in den Pflanzen dagegen bestehen nach ihm? in der Er- méglichung der Assimilation der Phosphorsiure bei der Bildung von Lecithin und Nucleoproteiden, weil die Phosphorsiure am leichtesten von allen in den Pflanzen sich findenden Phosphaten aus dem sekundiren Magnesiumphosphat abgespalten werden kann. Er hatte die merkwiirdige Giftwirkung von oxalsauren Salzen, welche nur fiir héhere Thiere und einige Pflanzen bekannt war, nicht nur bei verschiedenen Phanerogamen,* sondern bei den héheren Algen und niedersten Thieren beobachtet. In des Wirkung der Oxalate auf niedere Wasserthiere liessen sich nun grosse ° ~ Unterschiede erkennen, der Tod trat bei einigen Arten (Asseln, Copepo- 2 Archiv fiir Anatomie und Physiologie 1got. 9 . - > A£.2 ® Flora 1892, 368-394. 2 Von Interesse ist hier die Thatsache, dass jedoch geringe Mengen léslicher Oxalate normalerweise n manchen Pflanzen vorkommen kénnen, 146 M. Toyonaga: den, Rotatorien) sehr bald, bei anderen (Wasserkafern, Wassermilben und Nematoden) aber weit spiater ein, was wohl mit der verschiedenen Schnelligkeit zusammenhiangen mag, mit der jene Salze zu den wichtigeren Organen vordringen kénnen. Ino.5 proc. Lésungen neutralen Kalium- oder Natrium-Oxalats sind Asseln, Copepoden und Rotatorien in 30-50 Minuten todt, dann folgen Egel und Planarien, hierauf Insektenlarven und Ostracoden, wahrend nach 24 Stunden noch leben Wasserkafer, Wassermilben’ und einzelne Nemato- den. In einem Controlversuch mit neutralem weinsaurem Kali lebten fast alle jene Organismen noch nach 24 Stunden, viele noch nach mehreren Tagen. In einer 0.1 proc. Lésung neutralen oxalsauren Kalis starben Asseln, Copepoden und Rotatorien nach 3-4 Stunden, kleine Planarien nach 3 Tagen und Ostracoden waren darin noch nach 8 Tagen lebendig. Fadenlagen, wie Zyguema, Mougeotta, Vaucheria, Sphaeroplea, Cla- dophora, Oedogonium sterben binnen 24 Stunden unter Verquellung der Chlorophyllkérper in einer 0.§ proc. Lésung von neutralem oxalsaurem Kali ab. Bei Spirogyren lasst sich sehr gut beobachten, dass zuerst der Zellkern angegriffen wird. Derselbe quillt in einer 0.5 proc. Lésung nach ciniger Zeit auf und wird 6fter zu einem unregelmissigen zackigen Gebilde. Liisst man aber eine 2 proc. Lésung auf diese Algen einwirken, so gewahrt man schon nach 5 Minuten, dass die Kerne sich auffallend stark contra- hiren und nach 1o Minuten kein einziger Kern mehr intact geblieben ist. Das Cytoplasma ist allem Anschein nach noch véllig unverletzt, doch erholen sich die Zellen nicht wieder, wenn sie nach 10 Minuten wieder in kalkhaltiges Quellwasser zuriickversetzt werden, die Zellen sind nach 24 Stunden in allen Theilen abgestorben. Der Kinfluss der 2 proc. Oxalat- l6sung macht sich bei den Chlorophyllbindern der Spirogyren in ca. 30 Minuten geltend, wobei eine Verinderung der Conturen durch Verquell- ung sichtbar wird.? 1 Wassermilben starben erst nach 20-22 Stunden in einer 1 proc, L.Gsung oxalsauren Natrons. 2 Bei Controlversuchen mit cbenso starken T.ésungen von schwefelsaurem oder weinsaurem [Kali blieben jene Ercheinungen aus, Ueber die Vertheilung des Kalks im thierischen Organismus. 147 Von Wichtigkeit ist es, bei der Thierftitterung solche Pflanzen als Nahrung auszuschliessen, welche lésliche oxalsaure Salze in geringen Mengen enthalten ; es wird einerseits der lésliche Kalk im Verdauungs- tractus ausgefallt und durch den mangelhiaften Kalkgehalt entwickelt sich Knockenbriichigkeit und andererseits treten durch Resorption ins Blut andre krankhafte Erscheinungen auf, die raschen Verfall und Tod herbei- fiihren. In den Riibenbliattern ist z.B. die Oxalsiure theils an Natron, theils an Kalk gebunden. Als Endresultat vieler Versuche ergab sich: Oxalsiure enthaltendes Futter ist in geringer Menge, wenn es nur kurze Zeit gegeben wird, als unschadlich auzusehen. Werden jedoch die Bedingungen fiir die Unschadlichkeit des Futters (Zusatz von Calcium- carbonat) nicht erfiillt, so entwickeln sich die Symptome der chronischen Oxalsiéure-Vergiftung, wobei zuniichst die Nieren und Knochen in Mit- leidenschaft gezogen werden. Diese allgemcine Giftwirkung neutraler oxalsaurer Salze bei niederen, sowohl wie héheren Thieren erklart sich am einfachsten, wenn wir die bei Pflanzen sich ergebenden Schliisse, dass die Zellkerne zu ihrer Organi- sation Kalkverbindungen von Nucleoproteiden bediirfen,' auch auf die thierischen Zellen anwenden. Loew hat den Satz aufgestellt: ,,Je grdsser die Zellkernmasse in einem Organ, desto mehr Kalk enthalt sie.” Dieser Satz hat aber noch keine Beriicksichtigung bei den Physiologen gefunden. Vergleicht man jedoch bei verschiedenen thierischen Organen den Kalk- gehalt mit der relativen Grosse der Zellkerne, so findet man eine auffallende Uebereinstimmung mit jener Folgerung, soweit die bis jetzt vorliegenden Analysen Material zu Vergleichen liefern. Leider sind aber manche Organe noch gar nicht im Bezug auf ihre Mineralbestandtheile quantitativ untersucht, wie z. B. die Lunge, die peripheren Nerven, die Nebenniere, die Speicheldriisen und die Hoden: Von einigen Organen sind erst in neuester Zeit Aschenanalysen bekannt geworden. Aus dem obigen Satz wiirde z. B. folgen, dass Driisen kalkreicher scin miissen, als Muskeln, weil sie gréssere Zellkerne haben, ferner dass Muskeln niederer Thiere mehr Kalk enthalten, als die der Warmbliiter, 1 Ausgenommen sind nur die niedersten Algen- und Pilzformen, 148 M. Toyonaga: da erstere ebenfalls gréssere Zellkerne besitzen. Loew! hat einige von Katz? vor einigen Jahren publicierte Analysen dcs Muskelfleisches héherer und niederer Thiere verglichen sowohl unter sich, als auch mit dem vorliegenden Analysenmaterial von Driisen und fand seine Folgerung bestitigt. Wir wollen im Folgenden simmtliche Analysen von Kats beriicksichtigen. Dieser Autor fand in 1000 Th. frischen Muskels von Warmbliitern : Calcium. Menschenfleisch se pl hilags Gah, gia cis eRe eae Schweinfleisch ...) ««s .-,2 o=ce stun aoe Geen Gee ee ene ee Kalbfleisch...5 e302) “sostce cote cee eee Hirschfleisch., ~ .0. -<2«,. paid) Jeol Geis See ee) Kaninchenfleisch -... “ .... .c¢ (<8 See) eee Hundefleisch ...:, ...., ns.=\s25) [esc acne 2 Katzenfleisch « cis: 20 ) odie aged, gene a Hiilinerfleise lt): .3° ad% go00, <2< 0 peieeeee ee ee Durchschnitt ~ ..; 0. CR aoe) eee Diese Zahl ist etwas héher als die von Awnge* schon frither gefundenen Zahlen 0,086 und 0,072 CaO, entsprechend 0,061 und 0,053 Ca. Anderer- seits fand Ovdtmann in der Leber, dieser gréssten Driise der Siugethiere, 0,284 Theile Calcium in 1000 Theilen des Organs oder nahe 33 mal so viel als jener Durchschnittgehalt beim Muskel betrigt. Bei Kaltbliitern fand Katz in 1000 Theilen Muskel: Calcium. beim ‘Froschyi.. it 2b a, ee Schel lfisclt + sctyetvehsese me aet! Gem, oie eee ee Aa. 4 «cds wae eae aie oie tee a st ‘ Hecht sex. szegeleas opin een lade o esa) |) eee Durchschnitt eh Bae 6, 2 re 1 ‘The physiolog, Role of mineral Nutrients, Bulletin No, 18, U.S. Department of Agriculture, Washington 1899. 2 Jahresbericht f, Thierchemie 1896, p. 479. % Zeitschrift f, physiolog, Chemie 9, p. 60, Ueber die Vertheilung des Kalks im thierischen Organismus. 149 Wir finden somit, dass der Kalkgehalt der Muskeln von Batrachiern und Fischen weit grésser ist als der bei den Muskeln von Siaugethieren, in Uebereinstimmung mit obiger Folgerung. Fir den Magnesiumgehalt ergiebt sich umgekehrt, dass derselbe bei den Muskeln von Siugethieren grésser ist als bei denen von Batrachiern und Fischen ; namlich in 1000 Th. frischer Substanz : Magnesium Menschenfleisch ie mes oth .. -O:- 2116 SUneeMCISC Ms ee has cen ew tas bs. 0,2823 PecaPCISCI ew eten Pearse ans see oo 0,2434 PPO ISCIEN ee fe ese Ne ee vee wes tes |O,3044 PE CISCI Ran ce ke ek ee es ess 0;2006 PAOIGMCHIICISCIN ig fe. fos cae cee a0, ) oe. O,2869 ee een ee i es aa ok wae «-- O;2370 PeeseMMCISCUMET AN Gas Ges) usa seg cess 4s. , 0,2863 EegeumetnC Meee OP UE ROWER Ghat) ace sec ode ves, O,3713 PG Se Ot sehen rds e! Wied Base “aes: | a0 02793 In Uebereinstimmung damit fand schon Bunge fiir 1000 Theile Fleisch 0,412 und 0,381 Magnesia, oder 0,249 und 0,230 Magnesium. Vergleichen wir hiermit den von Katz gefundenen Magnesiagehalt der Muskeln von Batrachiern und Fischen, namlich: In 1000 Theilen: Magnesium Eee ereroON an i ek os ees tk, 2S ts | 0, 2353 Schellfishfleisch RC eM Feat! isha | cin, 0},2670 ees ey ct bc es es 0,8 782 PiOMEISC DWM GLE lat akO7 GANG Tag aes civ. ws. O,3TO2 PRIS NIECA NtE Pvt em wee aces vas) ve O,2227 so sehen wir, dass der Magnesiagehalt hier geringer ist als dort, wenn auch der Unterschied fiir Magnesium geringer ist als der gegentheilige fiir Calcium. Ozdtmann* fand auf 3,62 Theile Kalk in der Leber der Sauge- thiere nur 0,19 Theile Magnesia, oder auf 0,284 Theile Calcium in 1000 1 Citiert von Haddidurton, S, 538 seiner chemischen Physiologie. 150 man M, Toyonaga: ~ Theil Organ nur 0,017 Theile Magnesium. Es ist somit der Kalkgehalt weit grésser als der Magnesiagehalt, wahrend beim Muskel umgekehrt der Kalkgehalt geringer ist als der Magnesiagehalt. Gossmann! fand ahn- liche Zahlen fiir die Pancreasdriise und Niere wie Ozdtimann fir die Leber. In 1000 Theilen Organ sind enthalten : Pancreas Niere : Or eee p ee reece as Decaag Or 015 0,1184 Rind 4 Mgte:c scm Seer are (O08 S8 0,0410 Cases, bie ee oes 0,2008 Mensch } se (Me ct es eu toe: ees sarees 0,0472 Seither sind weitere Analysen einiger thierischer Organe erschienen. So hat Liueng die anorganischen Bestandtheile der Pancreasdriise von zwei alten Frauen quantitativ bestimmt und fand 2,569 Calcium und 1,48% Magnesium (wahrscheinlich in too Theilen® der Asche). Azbaut? hat ferner die Milz in dieser Beziehung untersucht, wobei er jedoch die Pulpa vom Bindegewebe durch Auspressen trennte. Die Pulpa betrug 41,5; 60,4 und 73,0% der getrockneten Organe. Er erhielt folgende Werthe auf Trockensubstanz bezogen : Calcium, Magnesium. Ga 7 Me: I I] It Il. Tu IU, I I Il 0 07 O07 0/7 oO of : P 7 70 4/9 70 70. 79 Ganze Milz 0,12 0,153 0,141 0,054 0,058 0,055 2,38 2,62 2,56 Pulpa 0,247 0.183 0,158 0,070 0,082 0,067 3,51 2,24 2,36 Bindegewebe 0,046 0,105 0,098 0,026 0,025 0,023 1,76 4,34 4,26 Auch A/oy* hat gefunden, dass in Milz, Pancreas und Niere, ferner in Knorpel und Bindegewebe der Kalkgehalt grésser ist als der Magnesia- gehalt. Derselbe fand, dass in Gehirn und Muskel das Verhaltnis es M kleiner als 1 ist, wahrend er fiir Milz, Pancreas und Niere die VWerhalentese fand 6.79; 4.03; und 1.84. Adoy scheint keine Kenntnis von den oben bereits discutierten Beziehungen zwischen Kernmasse und Kalkgehalt 1 Jahresbenicht f, Vhierchemie 30. 5, 419. Ibid, S. 386. 3 Thid. S. 492. i] ¢ Jahresbericht f, Thierchemie 30, S. 492. Ueber die Vertheiiung des Kalks im thierischen Organismus. IST gehabt zu haben, sonst ware er von seinen Resultaten weniger tiberrascht worden. ; Was meine eigene Untersuchung betrifft, so habe ich zunachst die weisse und graue Gehirnsubstanz separat auf Kalk- und Magnesiagehalt untersucht. Diese beiden Gehirnsubstanzen unterscheiden sich bekannt- lich sehr bedeutend in mehreren Beziehungen. Die graue Substanz besteht aus Nervenzellen mit einem wohl entwickelten Nucleus, wahrend die weisse Substanz lediglich aus Nervenfibrillen besteht, welche aus Nerven- zellen hervorgehen und nicht wie diese wohl ausgebildete Kerne haben. Obwohl zahlreiche Untersuchungen beider Substanzen vorliegen, was die organischen Bestandtheile betrifft, existirt doch noch keine Analyse der Asche von jeder dieser Substanzen fiir sich. Trotzdem mussten solche Analysen von hohem Werth sein, mit Riicksicht auf die grosse Zellkern- masse der grauen Substanz. Der totale Aschengcehalt des Gehirns variiert nach verschiedenen Autoren von 0,1 zu 1,0%, und nach Geoghehan! enthalten 1000 Theile Total- Gehirn: Ganze Asche Kk Na Mg Ca Cl POx COz) [HEPO,)- 2,95-7,08 | 0,58-1,77 | 0,45-1,11 |0,017-0,072,0,006-0,022) 0,43~-1,32 | 0,95-2,0I | 0,24—-0,79 | 0,01-0,09 Wahrend Geoghehan das Lecithin und damit eine betrachtliche Menge Phosphorsaiure vor dem Einaschern des Hirns mit Aether entfernte, somit auch weniger Totalasche erhielt als andere Autoren, fiihrt auch das directe Einaschern des Hirns zu Fehlerquellen, da viel Phosphorsaure aus dem Lecithin frei wird, die nun wahrend des Einascherns mit der gliihen- den kohligen Masse zu lange in Bertihrung bleibt.2. ‘Ich habe, um Verluste zu vermeiden, und den Verbrennungsprocess zu erleichtern, bei dem Einaschern eine bestimmte Menge Soda (meist 5 g.) zugesetzt und die Menge nach dem Veraschen und Wiegen wieder abgezogen. Allein auch diese Methode liefert keine fehlerfreie Bestimmung der Gesammt- 4 Citirt in Haddburton’s Chemical Physiology 1891, p. 517. Nach Z. f, physiol. Chem, Bad. 1. S. 335: 2 Geeghehan zeigte tibrigens, dass diese freie Phosphorsiiure auch Carbonate zersetzt, welche beim Einaschern des Gehirns erhalten werden, Ein von Lecithin becfreites Gehirn liefert namlich nach hm eine Kohlensiiure haltige Asche, 152 M. Toyonaga: asche, weil wahrend des Veraschens, ein Theil der zugestzten Soda in Folge der Bindung der Lecithinphosphorsiure Kohlensaure verliert. Woch mir kam es lediglich auf die Bestimmung der Kalk- und Magnesia- mengen fiir 1000 Theile frischen Gehirnes an und ich habe desshalb das Problem, eine méglichst einwandfreie Bestimmung der Gesamtmineral- stoffe im Gehirn zu liefern, noch einstweilen bei Seite gelassen. Ich habe sowohl das Gehirn vom Pferde als auch vom Kalb unter- sucht, nachdem ich die weisse Substanz so gut als mdglich von der grauen trennte.!_ Eine frisch abgewogene Portion wurde zunachst im Wasserbade eingetrocknet, zuletzt im Luftbade. Nach Zusatz von Soda wurde ein- geaschert, nach Extraction mit Wasser die Asche weissgebrannt und nun Kalk und Magnesia in itiblicher Weise bestimmt. In gleicher Weise bestimmte ich ferner diese Basen in den peripheren Nerven und der Lunge des Pferdes. Die Resultate sind aus folgender Tabelle ersichtlich. ‘Angewand- Kalk. , Magnesia. ites Gewicht) 446 = an frischer}| ~~ Sr aiae g, In der In 1000 Inder | In 1000 eae Be Asche |Thl.frischer} Asche |Thl.frischer oP g. Substanz, g. Substanz, Graue Hirnsubstanz. $9,50 3 0,0329 0,368 0,0227 0,254 Kalb | Weisse Hirnsubstanz, 158,00 0,0692 0,058 0,0094 0,060 Graue Hirnsubstanz. 23,970 0,0261 1,089 O,OI1TI | 0,463 Pferd - Weisse Hirnsubstanz, | 68,106 5 0,0045 0,052 0,0135 0,203 Periphere Nerven des Pferdes, 39,720 0,0315 0,794 0,0239 0,602 Lunge des Pferdes, | 61,050 0,0313 0,513 0,0274 0,449 Is hat somit die Analyse in Analogie mit den obenerwahnten Verhilt- nissen auch hier fiir die graue Substanz mit ihren zahlreichen Zellkernen einen grésseren Kalkgehalt ergeben als fiir dic weisse, an Zellkernen weit 1 Die graue Substanz macht 37,7-39.09¢, die weisse 61,0-62.39% des Gesammthirns aus, Ueber die Vertheilune des Kalks im thierischen Organismus. I airmere!, Ferner tiberwiegt in der grauen Substanz der Kalkgchalt iiber den Magnesiagehalt, wahrend in der weissen das Umgekehrte der Fall ist. Der Nucleingehalt ftir die graue und weisse Substanz ist noch nicht separat chemisch bestimmt worden. Nach Faksch enthalt des Totalhirn in 1000 Theilen nur 3 Theile Nuelein, nach Geoghehan gar nur 1,34-1,62. Wenn nun auch die Grundlage dieser Berechnungen keine ganz sichere ist, so bleibt soviel jedenfalls richtig, dass der Nucleingehalt nur gering ist. Damit wiirde also auch der relativ geringe Kalkgehalt des Totalhirns stimmen. Doch diirfte immerhin die von Geoghehan hiefiir gefundene Zahl 0,006 Ca auf 1000 Theile Gesammthirn auf einer mit Fehlern behafte- ten Analyse beruhen. Magnesia und Kalkgehalt des Gehirns scheinen betrachtlichen Schwankungen unterworfen zu sein; wahrscheinlich tiben Art und Alter der Thiere, sowie der wechselnde Fett- und Wassergehalt? hierauf, wie auf den Procentsatz an Gesammtasche einen wesentlichen Einfluss aus. Wie schon erwihnt, fanden verschiedene Autoren den Aschegehalt des Gesammthirns zu O.I-1.0 Procent; ferner wurde das specifische Gewicht der grauen Substanz von 1.029-1.039 schwankend gefunden. Bei krankhaften Zustin- den ferner werden weit gréssere Schwankungen eintreten kénnen. Um nur ein Beispiel zu erwahnen, kann bei Erkrankungen von Blutgefassen (Aorta) der Kalkgehalt bis zu dem 20 fachen des normalen ansteigen.* 1 Auch wenn wir den verschiedenen Fett- und Wassergehalt beriicksichtigen, bleibt dieses richtig. Es ergibt sich fiir too Theile trockne fettfreie graue Substanz beim Kalb =0o,.2g0 Thi, CaO; ditto weisse Substanz 0,075 Thl. CaO. 2 In Hammarsten’s Lehrbuch der physiologischen Chemie, III Auflage, S. 353, heisst es: .,die Menge des Wassers im Gehirn ist grésser bei jiingern Individuen als bei Erwachsenen.” Wie aus der darauf folgenden Tabelle ersichtlich ist, gibt es Ausnahmen. 3 Gaszert, Jahresbericht fiir Thierchemie, 1900. S. 511. 154 M. Toyonaga: Ueber die Vertheilung des Kalks im thierischen Organismus. Zusammenfassung. In Uebereinstimmung mit der von O. Loew gezogenen Folgerung, dass der Kalkgehalt mit der Masse der Zellkerne wichst, steht das Resultat meiner Untersuchung, dass die graue Hirnsubstanz relativ kalkreicher ist, als die weisse. On the Digestive Power of the Intestinal Canal, BY S. Sawamura. After the experiments of Thtry and Quince the digestive function of the -main part of the lower intestins was regarded to be of minor importance, but recent investigations disproved this view. Besides the accelerating action on the enzyms of the pancreatic juice observed by Schepowalnikow ' the variety of the enzyms found in the intestinal juice is of special interest. A diastatic enzym was observed in the human intestinal juice by Demant,? in the small intestines of the swine by Browz and Heroz,* in the small intestine of the dog by A7viiger,* Griinert,’ and Schepowalnikow,® and in the coceum and colon of various animals by Ludw¢g Vella? On the other hand, an énzym acting on inulin was proved to be absent in the human intestinal juice by Demant.8 Sucrase was shown to exist in the human intestinal juice by Demant,® in the small intestines of the swine by Brown and Aeron, in the intestines of the rabbit by Pavy,?! in the coecum and colon of various animals by Ve//a,!? in the small intestines of the dog by Griinert,13 Kriiger,14 and Bastianelli,+*® and in the small intestines of man by J/iura.1® Maltase was observed in the small intestines of the swine by Arown and Heron,1* in the intestines of 1 and © Jahresbericht ftir Tierchemie vol. XX, XT. p. 370. 48 and ® ibid. vol. IX. p. 222, SeeoPange=? abid.. vol. X. p.-77. 4 and 14 ibid. vol, XX. VIII. p. 337. 5 and 13 ibid. vol. XXI. p. 273. 7 and 18 ibid. vol, XV. p. 297. 1a ~ + ibid. vol. XIV. p. 294. 46 Jahresbericht fiir Agrikulturchemie. 1890. p. 523. 16 Jahresbericht fiir Tierchemie, Vol. NXNV, p. 288. 156 S. Sawamuras the rabbit by Pavy,! in the coecum and colon of various animals by Vella,? and in the small intestines of dogs and cattle by Pamutz and Vogel ;* Lactase, in the intestines of various young animals by Rémann and Lappe,4 Pautz and Vogel,®> Portter,® Orban’ and Wetnland.® Pautz and Voge/® found an enzym which acted upon raffinose in the intestines of the dog and cattle. As to the occurrence of cytase, the opinions differed, but it was finally decided in the negative. As to the behavior of digestive juices to various hemicelluloses, our knowledge is still imperfect. Hauber!° in Voits laboratory, in experi- menting with a dog to investigate the digestibility of mucilages, found | that they were digested and absorbed. He also observed that the elycerin-extracts of the stomach and pancreas of the same animal trans- formed mucilages into sugars, while ptyalin had no such effect. Stove and Foues,' and Lindsey and Holland!? observed the digestibility of pentosan by the rabbit, Wezske15 by the sheep and Kéntg and Reznhartd by man.?445 But V2/sou!® observed that no sugar was formed from lichenin by digesting it with the gastric and pancreatic juices at 36°C, for 24 hours. As to the digestion of fat by the intestinal juice Ve//a'* observed that it is only emulsified by them, while the absence of a special enzym was 1 Jahresbericht ftir Tierchemie. vol. XIV. p. 294. = and 45 ‘Ditto. vol. XV. p. 207. 3) 5-and 2 Ditto; voll ex lV pe sod 4 Ditto. vol. XXV. p. 286. 6 Ditto, vol, XX. VIM. p. 723 7 Ditto, vol. XX. IX. p. 384. 8 Ditto, vol. XXIX. jp. 382. 10 Ditto. vol. IV. p. 375. 41 Jahresbericht ftir Agrikulturchemie, 1893. p. 373. t2 andl 43) Ditto vol enso sane oie 14 Ditto. 1893. p. 53. 45 Chemisches Central-Blatt. 1902. vol. 1. p. 673. 16 Tferbivora can digest pentosan better than omnivora, and as it is always hydrated by digestion, there must be present a special kind of cytase in the intestines, Zeitschrift fir physiol, Chem, Vol. 36. p .65-66. 17 Jahresbericht fiir Tierchemie, vol, XV, p. 297. On the Digestive Power of the Intestinal Canal. I wt N proved by Demant! in the secretion of the intestinal tract, and by Schepowalnikow? inthe intestines of the dog. A proteolytic enzym was proved to be present in the intestinal secretion of the dog by Griinert) Gachet,* and Schepowalnikow,® but Demant® (in the human intestines) and Kriiger® (in the intestines of the dog) questioned its presence. Although various kinds of enzyms were hitherto observed in the intestines, little attention has been paid to the special parts of the intestines in which they are produced. Since both, Lezeberkithn’s and Brunuer’s glands, decrease in numbers gradually towards the rectum and especially in some animals the latter glands are found only in the part of the small intestines near the stomach, there must be some difference in the production of enzyms between the small and large intestines.’ I have directed my attention to the occurrence of enzyms that can saccharify mannan and galactan in the different parts of the intestines. In Japan a preparation consisting chiefly of mannan and derived from the root of Conophallus Konyaku and further the common agar are consumed generally by the people and since those hemicelluloses are digested we have to infer the occurrence of mannase and galactase in the intestines. The question seemed to me of some importance whether such enzyms are produced in all the various parts of the intestines. I compared in this regard the small intestines, coecum and colon, testing these three parts at the same time separately upon the production of the above enzyms as well as of others also. The intestines serving for these experiments were taken from a horse, well washed with water, and exposed to the air fora day. 50 ers. of the small intestines, coecum and colon cut into pieces, were digested with about five times their weight of dilute alcohol (359%) for a week. The filtered alcohol extracts were precipitated with ether-alcohol, the precipitate 2 and 5 Ditto. vol, XX. IX, p. 379. 3 Ditto, vol. XX. p. 273. : Ditto. vol, XX, VII. p. 377. 7 Ditto, vol. XXVIII, p. 357. ® LEllenger, Uistologie der Taussiiugethiere, p, 694. 158 S. Sawamura: dissolved in a 0.25% solution of sodium carbonate, and with the addition of some thymol served for the- following experiments with crude fibrin, neutral olive oil, starch solution, cane-sugar, mannan and cellulose (filter- paper). The results observed after three day’s digestion at 36°C were as follows :— Iéxtract from the small Materials used Eads eee Extract from the coecum Extract from the colon intestines x Kibrin A little attacked Not dissolved Not dissolved “ae Oil | No acid ert No acid reaction No acid reaction ie Mauch sugar Some sugar ; Litile sugar ; i eet Sugar reaction positive Sugar reaction a Sugar fees positive wi Canesugar Inverted ; Not inverted Not inverted : " ihe No sugar « No sugar No sugar The identity of the sugar produced with mannose was proved by the production of the difficultly soluble characteristic phenylhydrazon. The enzym-production is therefore not quite the same in the various parts of the intestincs. As to the enzym which acts upon mannan,—the mannase—it exists in all three divisions of the intestines. A second experiment was perfermed with the intestines of a swine. The extracts were prepared by digesting respectively 50 grs. of the duodenum, coccum., colon and pancreas (as a control) in 200 cc. of 20% alcohol for a week. The experiments were carried on with the addition of some thymol. The results observed after digestion at 36° C. for three days were as follows!:— 1 ‘The extract was proved to be free from any reducing sugars by testing with Zcding’s solution. On Digestive Power of the Intestinal Canal. 15 Ne) Fibrin Oil Extract from the duodenum. Not attacked No acid reaction Extract from the coecum, Not attacked No acid reaction Extract from the colon. Not attacked No acid reaction Extract from the pancreas. Dissolved completety Acid reaction ~ Starch Sugar reaction positive Sugar reaction positive Sugar reaction positive Sugar reaction positive Mannan Sugar reaction positive Sugar reaction positive Sugar reaction positive Sugar reaction positive Galactan Sugar reaction negative Sugar reaction negative Sugar reaction negative Sugar reaction negative Saccharose Tnverted Not inverted Not inverted Not inverted The sugar produced from mannan was proved to be mannose by test- ing with phenylhydrazine. As will be seen in the above table proteolytic and lipatic enzyms were absent in the intestines, while diastase and mannase were present in all the parts of the intestines.! Contrary to our expectation the absence of galactase was proved in this experiment.” We may conclude from these result as follows :— 1. In the intestines and pancreas of the higher animals, there is present manuase besides the other enzyms already observed. 2. The enzym-production is different in the small intestines, coecum and colon, the most notable being the absence of swerase in caecum and colon in the case of the horse and swine, and of ¢rypséz in the case of the horse. 1 Diastase was also present in the rectum, 2 A mucilage consisting of mannan, galactan,. and araban was digested with the extract of the intestines of the horse and swine, and it was found that some arabinose was formed in both these cases, as Was proved by phloroglucin and hydrochloric acid in the alcoholic extracts of the digested mucilage. 160 S. Sawamura: On Digestive Power of the Intestinal Canal, 3 Cytase was not observed in this experiment. 4 The function of the intestines in digestion must be regarded to be very important, since they secrete enzyms such as swcrase that are not contained in the pancreatic juice. On the Action of Manganese Compounds on Plants, BY O. Loew and S. Sawa. With Plate XII. The almost universal occurrence of manganese in the ashes of plants is a fact known long since, but as plants have been raised in water-culture to perfection in absence of manganese, this element is considered to be without any intrinsic value for the life of the plants. But it is nevertheless of interest to note the relatively large quantity occurring in the plants, exceeding often that of the related and so important iron. Thus, in the ash of beech leaves was. found in one case 11,25% Mn;Q, and only 1,07% Fe,O;.1. It was found in the most different organs, even in the’ pollen- grains,” further also in the ash of parasitic fungi feeding on the sap of trees. Young shoots and Ieaves are especially rich in it.3 It deserves to be pointed out that in one case mangano manganic oxid, Mn, O,, formed even the chief constituent of a plant ash. This being really an extraordinary case we mention the composition of the said plant ash. LT. Schréder* found on analysis of the ash of various parts of a pine tree (Pinus abies,) among other things the following result : Leaves Bark (Ab oe ere ewes i 1.805 % In 100 parts of pure ash ies Gene cia sie ery eS a 20.4696 INaSOS = os... EER 0.67 0.38 i A Cin VT oe, EE 14.72 1 IVolg’s Tables of Plant Ashes, I, p, 121. 2 Kamann found it in the pollen grains of the pine, Botan, Centralbl. 1898, 8 Fichard, Compt, rend ; vol 126, p. 550. * Forstchemische und pflanzenphysiologische Untersuchungen, Tharand, 1878. Also: Jahresbe- richt fur Agriculturchemie 1878. » \- ’ 4 4 q foz G. Leew and S$. Sawa: MeOs Si one Ponte! Oe. eee 7.14 Fe, O, = id, hea e eae ARO a eiegs S« oe, ee 3.61 Mn; OF. 22 ee Sea er ce eee 41,23 Ps Oso ee tbe ee ee GNOL Bt) lose ee eee 6.73 DOS tae chee eae GEO Sine le gens 2) Sen eee 2.69 Si. Oy = ..02 Se ees OF ck a. Als ee The manganese content, as Mn, Oy, for the dry matter of these leaves was therefore 1.089 and that of the bark 0.669, corresponding to 0.69 and 0.429@Mn respectively.— Animal organs contain much less manganese than vegetable organs. Rzche found only 0.5 milligrams Mn, O, in one kilo ef blood and according to other authors it is often completely absent. Wurzer (1833) observed it in the ash of the liver and teeth, Wetdenbusch in the bile, Horsford (1851) in urine, Pol/acce (1871) in milk and eggs, Maumené (1883) in hairs and bones, Péchard (1898) in molluscs, crabs, sardines, pigs blood and hens eggs. The general occurrence in animals forms apparently a contrast to the highly poisonous properties it shows on subcutaneous and intravenous injections. Eight milligrams of manganous oxid in the form of sodium-manganese citrate represents according to Kodert the letal dose per kilo body weight of a dog. On introduction into the stomach even large doses of manganese compounds prove harmless, on account of deficient absorption by the intestinal walls.? In regard to the behavior of plants towards manganese compounds, but few experiments have been made and these show, that manganese cannot replace the related iron in regard to the production of chlorophyll, and that manganous and manganic phosphate suspended in culture solutions can exert an injurious effect.2. Recently Gzg/éolz3 applied peroxid of manganese as an addition to various manures cn fields and observed in 1 Tn recent times manganese compounds commenced to play a réle in therepeutics, A manganese- iron-pepton preparation is frequently recommended for certain disorders, 2 Cf, Birney and Lucanus, Landw, Yersuchs-Stat, vol. 8, p. 128 and Wagner, ibid. vol. 13, p. 69 and 278, 3 Ann. Della R. Scuola Sup, di Portici 1g00. ‘The experiments were made with wheat, The peroxid of manganese was applied in the proportion of 1,14 ctw. per ha, Cf, also Centralbl, f, Agricultur- chemie 1902, No, 3. On the Action of Manganese Compounds on Plants. 16 some Cases a moderate increase, in others a decrease of the harvest. The result was not decisive, which will not create surprise since peroxid of manganese is a compound which hardly can be attacked and dissolved by the rootlets. The influence of manganese compounds in high dilutions has not been studied, although some action or other might be expected, considering the chemical properties of these salts, so closely related to the salts of iron, considering further the occurrence of manganese in the ash of oxidizing enzyms (Lertrand) and in that of certain nucleoproteids (A sd).} In order to observé the character of the injuries caused by manganese, young pea plants, 16-17cm. high were placed ina solution of 0,259 man- ganous sulphate but this concentration injured the plants within five days so considerably that no characteristic symptoms could be observed. Most leaves had lost their turgor, some had even perfectly dried up, and no trace of new rootlets became visible. The control plants, however, remain- ed perfectly healthy and had commenced to develop water rootlets. In the following experiment young barley plants 15—18 cm. high were placed in a 0.1 percent solution? of the same salt and kept ina heated room near a window. In this dilution of the manganese compound the injury develop- ed more slowly. After seven days, however, a gradual change from green to yellow was evident, and this phenomenon became very marked two days later. Some water roots had developed, although much smaller ones and fewer as in the control case. A checking influence of the manga- nese was quite evident. Onthe ninth day further observation was given up and comparative tests were made as to the color reactions upon oxidiz- ing enzyms. Five grams of the upper halves of the leaves were finely triturated with addition of pure quartz sand and gradual addition of 50 cc. water. This liquid had a weak acid reaction but this was still weaker in the control case. A portion of the colorless filtrate (f) was exposed for several hours to the air, whereby it assumed a reddish hue which was not observed in the control case. One cc. of the filtrate (f) was diluted with 20 cc. distilled water and five drops of a 2% alcoholic guaiac solution added * Bull. College of Agriculture, Tokyo ; vol. 4 No. 3. 2 Such data refer in this article to the anhydrous compound, not to the crystallized salt. 164 0 Loew and S. Sawa: whereby the blue color produced was much more intense than in the control case. A considerable difference in the intensity of the guaiac reac- tion for peroxidase after killing the oxidase by heating to 75°C., and adding some hydrogen peroxid was also observed. But still more striking were the differences in the tests with guaiacol and with paraphenylendiamine in presence of: hydrogen peroxid. One cc. of the filtrate (f) was diluted with 20 cc. of water and five drops of a 19% aqueous solution of guaiacol and three drops of dilute hydrogen peroxid added. A red-brown color of great intensity set in at once, while in the control case a much weaker coloration was slowly developed. A colorometric tést fifteen minutes after adding the reagents showed that in the latter case tlie intensity of color was less than one half that of the former. In an analogous manner the test with paraphenylendiamine hydrochlorid (to which a little sodium acetate had been added) and hydrogen peroxid was carried out. The sreen color in the control case was but half as intense as in the case of the manganese plants. The undeniable fact that the reactions on the oxidizing enzyms are more intense with the manganese plants than the control plants can give as also an account for the fading out of the green color of the leaves. Our result corroborates the statement of Bertrand! that the oxidizing enzyms act more powerfully in presence of manganese compounds than in their absence, and that iron compounds cannot produce an effect of the same intensity.? Theeffect of the manganese seems to be the same, as an increase of the oxidizing enzyms by certain stimulants secreted by parasit- ary insects (Aphides) and fungi. The yellow spots thus produced on leaves yield according to Albert I. Woods more intense reactions upon oxidase and peroxidase than an equal surface of the healthy leaves. Woods also observed a higher content of oxidases in etiolated shoots, than in normally green ones. 1 Compt. rend,, vol. 124, p. 1032. 2 There may exist cases in which oxidizing enzyms are not associated with manganese or iron but they will be less powerful in that case, Cf. also Savthow, Journ, Pharm, Chem, vol 11, p. 583 [1900.] 3 C, Bakt, II Abt. 5, S. 745 [1899.] ——— On the Action of Manganese Compounds on plants, 165 In our next experiments! the manganous sulphate was applied in a much higher dilution in the cxpectation to diminish the injurious effects toa minimum. At the same time the mineral nutrients were offered to the plants in the following proportions ; ei MMGAbe .caa. <6 eocs ewer te ee een 0.0495 PU eM eS MO SU PACE gs a sae os cle aye oe «2 os 0.01% ES SUL AL On uren aideyls eevee eee os ss = «0103 99 Monopotassium phosphate...............0.02% PERIIOMUUMSUP MALE. Los view eae wes. - 0.01% To one portion was added 0.01% ferrous sulphate (control solution), to another 0.02% manganous sulphate and toa third 0.01% ferrous sulphate plus 0,029 manganous sulphate. Shoots of barley and soy bean were placed in these solutions and kept near a window in a room whose temperature ranged from 4—12°C. during the first three weeks of observation. After a series of days the shoots in the solution containing manganese and iron jointly exhibited an increased growth. The measurement revealed even considerable differences (see Table); gradually however these shoots durued yellowzsh, their as- similation power was consequently depressed and in further consequence of this the decreased nutrition led toa relaxation in the speed of growth, as seen from the measurements of the soy bean plants. Experiment with Barle gy. Two shoots were placed in each of the solutions above mentioned March 27. 4 ye also have made experiments with, algae but they failed on account of development o parasites, 106 0. Loew and S. Sawa: Date of Measurement, cm. : Increase after 25 days. ea March 27 April 9 April 22 um A 34.0 46.0 5E5 51.4 B 28.0 47.2 55-5 46.5 Experiments with Soy bean. Three shoots were placed in each solution, March 25. Date of Measurement, cm. March 25. April 8. Apri! 22. April 30. May io. | \ 9-3 26.0 37.0 45.0 Mn.4B 10.2 22.0 25.4 : 40.5 |. 7.0 22.5 31.0 dead. 56.5 Mn + Fe 435 — | | | 51.5 | | EE | eee On the Action of Manganese Compounds on Plants. 167 The gradual increase in the speed of growth upto April 22 and the following diminution of that speed with the plants that received iron and manganese is still clearer seen from the average : March 25. April 8. April 22. April 30. May to. Mn+Fe 8.9 25.3 30.4 45.8 50.5 cm. Fe 8.5 2275 33.5 42.5 50.2 cm. From the yellowing of the leaves under the influence of manganese might be inferred that this is a strong poison for plants. But this conclusion would not be justified, since our further observations have demonstrated beyond any doubt that at summer temperature the plants are capable to overcome the yellowing effects of the manganese, if this is present only in small quantities. It appears that by the increased activity of the protoplasm a part of the dissolved manganese in the cells is transformed into insoluble compounds!, a process, that probably takes place in nature, when plants absorb manganese compounds from the soil. Thus it may be rendered possible that only such small quantities remain dissolved in the cells as can exert a beneficial effect. Experiment with Rice in Soil Culture. In the following experiment with rice in soil culture the yellowing was not observed at all. The soil came from the experiment grounds of our College of Agriculture. Each pot containing eight kilo air dry soil was manured with 16 g. superphosphate, 10 g. potassium carbonate and 16g. sodium nitrate. Pot I received no further addition. Pot II] was watered with 200 cc. of ao.1 per cent solution of ferrous sulphate, Pot IT with this solution and further with 2co cc. of a Or per cent solution of manganous sulfate. The seed was sown on May 24 and the number of shoots reduced four weeks later to seven about equally large ones. The rice was cut on Nov. 10 with the following result : 1 The observation of so (l.c.) that a nucleoproteid contained besides iron also manganese may furnish us a clue in the right direction, Z. 168 0; Loew and §. Sawa: T, Control, lhe sO; II, Fe SO,4+MnSo, Number of stalks 19 | 20 18 Average length 58.6 cm, 59-7 cm. 64.6 cm. Weight of straw ANG AC, 46.5 g. 48.7 g. Weight of grains 5.7 g. 7.0 &. 1,219; These numbers show that ferrous sulphate alone exerted a manuring effect, an observation also formerly made by others. Soils which contain iron ina difficult soluble condition will respondin this manner. Much more striking was here however the effect of the manganese. Although the number of stalks was smaller, the weight of straw and especially that of the grains was larger than in the control cases Iand JI. The stimulat- ing effect of the absorbed manganese was exhibited in an unmistakable manner. Nevertheless the inference would not yet be justified that every soil would respond in this manner. Probably many soils with a high natural fertility contain manganese in an easily absorbable condition, in which case a further supply of manganese salts would be of no avail.’ It would be of interest, however, if in the analysis of soils more attention would be paid to the manganese content the determination of which is often wholly neglected. Experiment with Pea in Soil Culture. The soil was here the same as in the last experiment. The main and the control pot held this time 2300 g. soil and each was manured with 3 g. sodium nitrate, 3 g. potassium carbonate and 4.6 g. common superphos- phate. On Febr. 21 fifteen seeds were sown and later on the young plants reduced to five equally large ones in each pot. The main pot received 1 One of us (Z.) has mentioned two years ago a case, in which tobacco plants showed no increase of the oxidizing power of the oxidases, after being irrigated with 0.1 per mille Mn SO, solution (Report No, 65 of the U. S. Dep’t of Agriculture, p. 22). — Pi On the Action of Manganese Compounds on Plants. 169 highly diluted solution of manganous sulphate on March 11 and 25; April 14, 21, and 28; and on May 6. The total amount of this salt was 0.036 g.; the first two times also o.oo1 g. ferrous sulphate dissolved in 100 cc. water was added. The plants commenced flowering on April 22, the ripe fruits were harvested on June 2. A photograph, taken on May 17 (see Plate XII.), shows the more luxuriant development under the influence of manganese very distinctly. The ripe fruits were weighed in the fresh state, further the peas isolated and weighed well dried at summer temperature. The straw was weighed in the air dry state. The results were as follows ; Manganese plants. Control plants. Weight of the fresh fruits 2 aR ae ee BO. athe tn Wheight of air dry seeds So eS Weight of the air dry straw ee en Ce OWT nica prainats These results leave no doubt of a stimulating action having been produced by manganese on the development of the pea plant, but the differences in the seed harvest were not so large as in the case with rice. The nodules on the roots were rather scanty with both the pea plants. Experiment with Cabbage. A small plot of 12,5 squ. Meter received 3 g. manganous sulphate (anhydr.) dissolved in 15 Liter water.!| The land had received the previous year twice barn-yard manure and had served for cultivation of barley and radish. This year it received only ammonium sulfate at the rate of so g. to 12,5 square Meter. Cabbage seed was sown ona part of this plot on April 25. Germination proceeded very slowly and many seeds failed to germinate. In the beginning of June, however, the difference in develop- 1 Two grams on April 24; rg. on May ar, 170 9, Loew and S. Sawa: ment of the plants on the manganese plot compared with the control plants became very striking. On June 14 numerous plant lice made their appearance and damage by beetles threatend, hence the plants were collected, the roots washed and by gentle pressure freed from the adhering water. All control plants larger than 12 cm. from the base of the trunc, thirtheen, weighed united in the fresh state, 56,0 g., while the same number of the manganese plants weighed 94,9 g. Hence a_ very favourable influence of the manganese is also here evident. The question may be raised: How is the remarkable stimulus cf growth by manganese to be explained? Can some light be expected by reviewing the characteristic properties of manganese compounds ? The fact that in many other cases it seems almost impossible to find an explanation for the stimulant action ought not to deter usin every new case from searching fora clue. Now, it is well known that light retards growth. This hitherto unexplained phenomenon forms a strange contrast to the great chemical work the light performs with the aid of the pro- toplasm in the chlorophyll bodies. .One and the same agency then increases the organic food on the one hand anc suspends the utilization of that food on the other. It is in the absence of light that growth proceeds and the products of the sun’s work are chiefly consumed. The absence of light has therefore the same effect as the presence of manganese. It seems as if under both these conditions a check is removed which the sun’s rays exert. This check might be due to the action of certaim noxious compounds produced in the cells under the influence of light. Such compounds (Hemmungs-Stoffe, Ermiidungs-Stoffe) are frequently produc- edin the course of the metabolism.1 It is the office of the oxidizing enzyms, as one of us (Z.) has suggested, to destroy noxious by-products of the benzene series, a view verbally expressed as follows?: “The writer’s view on this subject is that as the living protoplasm can oxidize carbohydrates and fat, but does not attack or attacks only with difficulty compounds of the benzene group, and, on the other hand, as 4 Cf, Ly, Reinitzer, Berichte d. botan, Gesellschaft, Vol. 11, p. 531 [1893]. 2 Report No, 59 of the U. S, Dep’t. of Agriculture, Washington 1899. p. 27. t ; On the Action of Manganese Compounds on Plants, 171 just the opposite takes place with the oxidizing cnzyms, it may be inferred that there exists between the protoplasm and the oxidizing enzyms a certain division of labor, the former oxidizing the compounds of the methan series and the latter those of the benzene series. The former provides for the kinetic energy of the cells; the latter destroys by partial oxidation noxious by-products. The oxidations in the former case are generally complete, but in this latter only partial.” If the checking compounds are gradually changed by partial oxidation, without being produced anew in darkness, we can understand the increased growth in the absence of light. Since, however, as we have seen above, the presence of manganese increases the oxidizing power of the oxidizing enzyms, the destruction of the checking compounds may be accomplished as quickly as they are formed and thus an explanation can be furnished why in presence of manganese the growth proceeds day and night while in absence of manganese only at night time. Future investigations will show whether this explanation is the correct one. It was in this connection of some interest to note that fungi show no enhancement of growth under the influence of small quantities of manganese although traces of other salts, as zinc salts (Richards) and copper salts (Oxo) can exert a stimulant action. A remarkable stimulant influence on the growth of Aspergillus under the influence of traces of sodium fluorid was also observed by Ono. These observations are in full accordance with Hitppe’s biological law. The different behavior of fungi towards manganese in this regard seems to indicate that the enhancement of growth of phenogams under the influence of manganese is not due to a direct stimulation of the protoplasmic activity, not to an irritation as it is observed by poisons in very high dilutions, but it must be accounted for by a different cause. The explanation just given by one of us (Z) would agree with this inference. The first experiments on the influence of manganese salts on fungi were made by AH. AMolisch.1 He concluded that manganese cannot enhance the development of fungi like iron salts can, the latter even being 2 Wiener Akad, Ber, October 1894. 172 @. Loew and S. Sawa: On the Action of Manganese Compounds on Plants. indispensable. ‘Recently Mr. Yakahashzt from this College made some further experiments in the same direction. As culture solution served a sake wort prepared from boiled rice by the action of Aspergillus oryze. Tothis wort was added after sterilization a sterilized solution of manganous sulfate, to a second flask ferrous sulphate and to a third ferrous and manganous sulphate jointly, while a fourth served as control. These liquids were infected with a pure culture of sake yeast while in a second series the flasks were infected with a trace of spores of Aspergillus oryze. Both series were kept in diffuse day light. After three weeks the fungus mass was collected on a weighed filter and dried. The result was as follows: Salts added. Weight of yeast. | Weight of Aspergillus. Manganous sulphate o.1 p.m. 0.764 | 1.122 Ferrous sulphate 0,1 p. mille. 0.778 1.394 Manganous and ferrous sulphates 0.05 p. mille each. 0.467 1.216 Control. 0.813 1.285 These results agree well with those of J/olzsch, there is no distinct stimulant action of manganese on fungi. It is true that the weight of the yeast failed also to show an increase by the ferrous sulphate, but here it must be born in mind that the original culture solution contained already some iron. Summary. Manganese exerts in moderate quantity an injurious action on plants, consisting in the bleaching out of the chlorophyll. The juices of such plants show more intense reactions for oxidase and peroxidase than the healthy control plants. Manganese exerts further a promoting effect on the development, still observable in high dilution, while the injurious effects disappear under this condition. It is probable that soils of great natural fertility contain manganese in an easily absorbable condition, and that this forms one of the characteristics of such soils. oF ae VHOMME, MGIC” (COOVEIER, WAVES V4 PLATE ALL. Table showing the influence of manganese on pea, I Manganese plant : I Control plant. To page 160. Ueber die Wirkung des Urans auf Pflanzen. VON Oscar Loew. Tafel XIII. ‘ Die Lichtempfindlichkeit der Uransalze’ liess es von Interesse erschei- nen, die Wirkung derselben auf griine Pflanzen zu verfolgen, da méglich- erweise Spuren von Uran im Chlorophyllkorn die Umwandlung von Licht in chemische Energie beférdern und damit die chemische Leistung vermehren konnten. Es wurde von Seekamp beobachtet, dass Bernsteinsaure unter Vermittlung von Uransalzen durch Licht in Propionsaure und Kohlensaure gespalten wird. Ferner bilden die Fluorescenzerscheinungen und die in neuester Zeit beobachtete Radioactivitaet? mancher Uranverbindungen interessante Beziehungen zum Lichte. Bis jetzt scheint nur ein einziger Versuch tiber die Wirkung von Uran- verbindungen auf Pflanzen ausgefiihrt zu sein und zwar durch Awof,* welcher Uranylphosphat als Aufschwemmung in der Niahrloesung ver- wendete. Kxop zog aus diesem Versuch den Schluss, dass wegen der grossen Schwerloeslichkeit des Uranylphosphats dasselbe ohne jede Einwirkung sei. Sorgfailtige Vergleiche mit Controlpflanzen scheint er nicht angestellt zu haben. Zwar gehdrt das Uranylphosphat mit zu den schwerléslichsten Phosphaten, doch geben Uransalze in einer Verdiinn- ung von o.1 pro mille keine Fallung mehr mit Monokaliumphosphat, sondern nur eine schwache Opalescenz. In solcher Verdiinnung kénnte t Bekanntlich werden Uransalze auch in der Photographie verwendet. 2 Radioactive Substanzen wirken im Dunkeln auf die photographische Platte ein und wenn nach Monaten diese Eigenschaft erlischt, so kann sie durch Belichtung mit Kathodenstrahlen wieder hervor- gerufen werden. Die Radioactivitaet scheint Beziehungen zu den Bequerelstrahlen und zur Phospho- rescenz zu haben, $ Jahresber, f. Agricultur- Chem, 1884, S. 139. 174 Oscar Loew: demnach wohl Uran auch in Gegenwart der Phosphate des Bodens von den Wurzeln aufgenommen werden, Zunachst wollte ich einige Daten betreffs des Giftigkeitsgrades! sammein, dann die Wirkung bei sehr grosser Verdtinnung beobachten. Auf junge Erbsenpflanzen wirkte Uranylnitrat schon in 3 Tagen sehr giftig ein, als diese in cine 0.2 procentige Loesung gesetzt wurden. Wurde diese Loesung bis auf 0.05% Uranylnitrat verdiinnt, und junge Zwiebel- pflanzen eingesetzt, so war nach sieben Tagen das Hauptblatt fast tiberall von der Spitze abwarts bis nahe zur Halfte der Lange gelb geworden und partiell verdorrt; die jiingeren Blatter wurden erst spater afficirt.2 Die Wurzeln hatten eine gelbliche Farbung angenommen, keine neuen Zweige, keine Wasserwurzeln waren erschienen, wahrend bei den Controlpflanzen in biosem Wasser dieses der Fall und tberhaupt das ganze Ansehen noch ein normaies war. Ganz ahnlich war die Wirkung auf junge Gersten- pflanzen von 12-18 cm. Hohe. Diese Pflanzen hatten versucht, neue Wasserwurzeln zu treiben, diese waren aber schon als kurze Stummeln wieder abgestorben, wahrend die Controlgerstepfanzen in blosem Wasser in sieben Tagen neue Wurzeln von bis zu 2 cm. Lange getrieben hatten. Wurden nun Erbsen- und Gerstenpflanzen in Nahrloesung gesetzt, welcher o.o1 per mille Uranylnitrat zugesetzt war, so liess sich selbst nach Wochen keine schadliche Wirkung mehr wahrnehmen. Nun wurde (Febr. 16) ein Topfversuch mit Erbsenpflanzen ausgeiiilrt, denen bis zur Beendigung der Bliitenperiode sechsmal je zwei Milligramme Uranylnitrat in 100 cc. Wasser gelést, gegeben wurde.? Die jungen Pflanzen im Haupt- und Controltopf wurden auf fiinf médglichst gleich grosse reducirt. Die Bliitenperiode dauerte vom 21 April bis zum 12 Mai. Am 17 Mai wurde eine Photographie aufgenommen welche auf Tafel XIII - reproducirt ist und die tippigere Entwicklung der Uranpflanzen deutlich 1 Auf Thiere wirken bekanntlich Uransalze sehr giftig und rufen Diabetes, Degeneration der Leber und Paralyse hervor. 2 Ein Vergleich mit Manganoxydulsulfat zeigte, dass dieses weit weniger schidlich wirkte als das Uranylnitrat. . 8 Dieser Versuch wurde gleichzeitg mit dem Manganversuch an Erbsen, und unter denselben Bedingungen angestellt (siehe den vorhergehenden Artikel). Ueber die Wirkung des Urans auf Pflanzen. | erkennen lisst. Auffallend war, dass die Uranpflanzen zur Zeit der Reife der Friichte neue Zweige aus dem Boden trieben, welche Bliten bildeten, was bei keiner der andern zur selben Zeit beobachteten Pflanzen der Fall war.! Am 2, Juni wurde geerntet, die Samen enthiilst und diese sowohl als das Stroh lufttrocken gewogen. Die Wurzeln hatten in beiden Fallen nur wenige Kndllchen entwickelt, doch die Controlpflanzen immerhin etwas mehr als die Uranpflanzen. Das Resultat der Wagung war wie folgt: Fiinf Uranpflanzen. | Fiinf Controlpflanzen. i 20,5 3 AG 510) ae L7AOres 10,7 g. Ein stimulirender Einfluss des Uranylnitrats mit Vermehrung nicht nur des Strohs sondern auch der Samen ist demnach zweifellos. Es ist dieses eine interessante Thatsache, doch verbietet der hohe Preis der Uransalze deren praktische Verwendung. Zugleich mit diesem Versuche mit Erbsen wurde unter ganz gleichen Verhaltnissen ein Versuch mit Hafer angestellt. Ernte am 3, Juli, Stroh und Samen (unenthilst) wurden im lufttrocknem Zustande gewogen, mit folgendem Resultat : Uranpflanzen., Controlpfianzen, SHOUCeAIMeS.. si. Gs. ek 11 9 GET Piya) sks ans tans 49.5 | 45.2 Korner mit Hiilsen ... ... ... 26.7 21.4 Die fordernde Wirkung des Urans ist somit auch hier unverkennbar. 4 Ausser mit Mangansulfat waren Pflanzen mit hoch verdiinnten Loesungen von KJ und NaF behandelt worden (siehe in diesem Heft Mr, ts0’s und Swsuki?s Artikel), ee ‘ - ae ~ 5 = . Pac, . ’ : ia a ; ‘ee Pd rr) 4 ‘ meres : by pe, ta a ct 2 ¥ Ped a : +s “a rw 1 ae - « ‘ “ a 7 ~ tel g s* | a A 3 ty “ “a iY es | pg we ; * « i ad - 37 am co t 4 a} “4 = ’ 4 b. . r fer : Ps P t { 1 . = i & : ‘ i bay be ; . 7 : i é} a ‘ re - : =) ole » = 5 ' » 4 — - 3 } ; a ‘ i f " “ . 7 : 5 7 “es < 4 - ’ oo a « ' ‘ 4 : ‘ { Poe Ps 4 ' A é : - Fi ee A 4 7 . oe H > 4 : ” om ‘ J F, 7 : . a F - “A y ? : f ‘ a : | : ! H " ‘ i > } 4 , 1 . i 4 7 2 aw ‘ ’ f *) 3 = 7 eE- . “1 j : i : : P - ,s i. sal i . 4 \ . 1 ; 4 i ‘in we ‘ +“ * 2 ¥ ; 4 ‘ . a { a . ? ’ ais gs ’ ‘ : i Seek, : Y , : . a \ - v * 3 ~ x ‘ ' i ‘ : - ; ‘ ; . ~ = ’ = .s * : as ao * : i vs a we : : is on . i a f - a oe — 4 ~ “ i ' u . . ve ' . - t-* ‘ ‘ f pe : > 3 ‘ ' ‘ : . _ i, \ . Ne ‘ ' ; ae , 2 - < } é = % ° , oy : a ; ¥ , ) ' t 4 : me x . " . ri Gok ] 7 : ’ “ws ‘ 4 iy . ‘ a en - re ot ; Domne - y 7 c a . ‘ se > ae y 5 ; ‘ - ws ‘s hd ’ . ’ - a “ 1 = . 4 3 . 4 4 ‘ i ° - ’ . ’ 4 . ‘ . ns! 1 . “y Lining ¢ } < : ! A M 5 " . ar ‘ ‘ . - : . : Fi ; a ‘ i : 2 a) : © Y : oe » ee : ; o ~ ’ H ‘ o 4 Pe) aire of i] 1 oe F A? : ‘ . . . 1 1 3 , ~ i . : ¢ A = . a ‘ a 1 < - K i Pre 4 5 . . * =e Fe - : fy : Ee ik ue = ey ic ‘ ; 4 “* Ap ; os —S ae . > : . ) *. JEN Mog, MCLE, (HOVLILR TAOME SW THGATELE. XALT, Die Tafel zeigt den Einfluss des Urans. I, Boden erhielt 0.012 nitrat IT. Controlpflanze. Zur Seite 174. é On the Physiological Influence of Manganees Compounds on Plants, With Plates XIV—XVI1I. It is a well known fact, that plants can develop normally in absence of every trace of manganese in water culture and further that manganese which is of frequent occurence in plants can not replace iron in the production of chlorophyll. But since certain metallic salts, as those of zinc, cobalt and nickel! can exert a stimulating effect on the growth of fungi when applied in high dilution, it seemed of interest, to observe also the action of manganese, so frequently present in the soils, on the growth of agricultural plants. Experiment with Radish. On Nov. 26, 1901, shoots of radish, 5-9 cm. high, were placed, two shoots in each flask, in the following solutions : A. 0,029% Mn SO, + trace of Fe SO, B. 0.029% Mn SO, + 0.02% Fe SO, (a. 9.02% Fe SO, Fach flask contained further the following nutrients : Ie cl eee Sue ues ues O.2 1) SE Se ee a TY "4 BO cs wy ag Es, owe OOK Y Lo a eres Tt (NH,). SO, see eee see eee eG 0.05.96 9 Cf, Ono, Journal of the College of Science, Imp, Univ,, Tokyd, Vol, XIT, part J., aso Rrcka and others, 178 K. Aso: In a second series, (a), (b), (c), the above solutions (A), (B) and (C) were diluted with ten times the volume of water, while the mineral nutrients were present in one fifth the quantity as in the first series. These shoots were kept in a cold room with a winter temperature of o°-6°C. After two weeks, the difference in development was very striking, as will be noticed from the accompanying photograph (Plate XIV) taken on Dec. 12. On Dec. 14, the following determinations were made: Number of leaves. Length, Fresh weight. { 4 11,2 cm, TNs Rhea sn 1.2 grm, | 4 10.0 ,, | 4 8.4 ,, ) Bice cee: : 0.65 5, 4 1 ss i ; ( 3 6:8: 5; Grae sancities 0.35 + | 3 61053 ] | 4 TES » 55 22 eR roeonane - 1.3 ” | 4 10.5) 3: ( 4 9.8 ,, De Sed scat 0.9 ” ( 4 8.8 Ae \ 3 8.2 ” Corn ieee 0.45 5, | 4 8.3 ” This result shows doubtless a most remarkable stimulating effect of the manganese. An undesirable feature was the gradual yellowing of the leaves, which however turned gradually again to a normal green, on transfering the plants to a heated room. A fungus now appeared on the roots, hence the experiment had to be terminated. Since Bertrand has repeatedly observed that the ash of oxidizing enzyms contains manganese and that in presence of manganese compounds the oxidizing effect of these enzyms is considerably increased, it was of interest to compare here those effects. Pieces of leaves of (a) and (c) of equal surface (5 x 7 m.m.) were well crushed in a mortar with addition of a On the Physiological Influence of Manganese Compounds on Plants. 179 10 c.c. of water. This extract (2 c.c. in each case) served for the following mtests ;+ 1. Upon addition of one drop of a 2% guaiac tincture, the blue color produced was more intense in the case (a), than in that of (c), and this difference became much more marked after one minute. 2. On addition of one drop of guaiac tincture and two drops of a 19% hydrogen peroxid solution, after killing the oxidase by heating, the blue reaction for peroxidase appeared, but the difference was not so striking as in the former case*with oxidase proper. 3. On addition of 1 c.c. of a 1% guiacol solution and two drops o hydrogen peroxid, the red reaction produced was more intense with (a) than with (c). 4. On addition of a few drops of dilute sodium acetate, parapheny- lendiamine hydrochlorid and hydrogen peroxid, a green reaction of much higher intensity was produced in (a) than in (c). 5. This difference was also noticed with the violet reaction which was obtained with tetramethyl-paraphenylen-diamine and hydrogen peroxid. Furthermore, two sections of equal size (4x6 m.m.) of the leaves of (A), (B) and (C) were ground with 15 c.c. of water. The tests (with the exception of 5) carried out as just mentioned, showed also in this case that the plant containing manganese (A and B) yield a juice which exertsa more powerful oxidative power than the plants without manganese (C). Experiment with Barley. On Nov. 26, barley shoots (7-8 cm. long) were placed in solutions of the same composition as in the experiment with radish. The influence of Manganese was here noticed not so early as with radish, but was very marked nevertheless, as seen from the following table, containing the determinations made on Dec. 14, and from the photograph taken, Dec, 12, (Plate XV). 1 Cf, also the paper of the writer, “On oxidizing Enzyms in the Vegetable Body,” in this Bulletin. 180 K. Aso: Number of leaves. Length. Fresh weight, | 2 19.5 €m. 7 Ne 8 eee _ | 3 ee? |) oes . ( 2 172), A SeE COCO f | Peds é { 2 TALI =; arn eee ; re \ 2 23-3 9» a Neneteo anecc 4 0.70 grm, ( 3 15.0 , ( 2 18) ee [asemeaetaae 5 OOS l 2 TO!St sss \ 2 | ES 5ess cs me | OSs, | 2 13.6 5, The series (A), (B), (C), was no further observed, since some shoots commenced to show injury, and demonstrated beyond a doubt as also did the above described experiments of Loew and Sawa, that barley in water culture suffers—at least at the low winter temperature—from the concent- ration of 0.02% manganese salt. But in regard to the series (a), (b), (c) in which manganous sulphate was applied in a concentration of only 0.002%, the observations were continued, after the plants were transfered (Dec. 14) from the cold room to a heated room. The following table shows the results: Jan. 9. Febr. 2. Bebra5. March 3. March 15. April 14. $e.) reat ef] rene Af rane N38 gee conn] cen Sere oe cm, pide cm, ey a ee a ee rn cm, a 8 27.0 4 27.1 4 30.5 8 35.5 b 7 20.2 3 20.2 3 26.0 7 52.5 c 8 18.5 2 22.0 5 23.4 8 Grek On the Physiological Influence of Manganese Compounds on Plants. 181 The fresh weight. Bebr. 2: March 3. April 15. a 4.0 grm. 9.7 grm. 16.8 grm. b Bea 5, G5, EZ. «5, 255 PP a ee Wea eee On Febr. 2, a difference in the color of the leaves was not yet noticed, but the yellowing of leaves in (a) was clearly observed on Fcbr. 15. The solutions were renewed on Febr. 3, March 3, and March 15. The roots in the manganese culture solutions turned gradually brown, so also did the lower leaves in (a), whereby the brown color was more especially con- centrated in certain points, which had also been noticed in the series (A), (B) and (C). On microscopical examination the membranes of the epidermis cells, and here and there also what seemed to be the nuclei proved to be deeply brown. For barley in water culture an addition of 0.01 per mille Mn SO, seems to be the highest concentration which is applicable without injury. The colorimetric tests for oxidizing enzyms were here made with equal weights of the fresh leaves of (a) and (c), and thus ascertained as in the case of radish that the yellowish leaves of the manganese plants gave reactions of higher intensity than the green leaves of the control plants. But the difference was here not so great as in the case of the radish shoots. Experiment with Wheat. At the same time at which the barley experiment was begun, one with wheat shoots, 6-7 cm. long, was started under the same conditions. The observations on the series (A), (B), (C), are contained in the following table: 182 K. Aso; Dec. 20. Febr,. 2. Number of = ae: Number of stalks, Length. Fresh weight. sfailicss Length, \ 3 230 CM. ( 0.g0 germ, 5 25.0Cm, 25.0 The leaves of this series gradually turned yellowish, and since one of the two plants in B and C died off, the observations on this series was discontinued, while those on the series a, b, c were continued until May 14. The data relating to this series are shown in the following table. Dec. 20, Pebrs ag: Fresh weight. No. of stalks. No, of stalks. Fresh weight. Fresh No. of Length, weight. stalks. Length. Length, germ. cm. erm, cm, grm, lee) 0.85 28.8 6.6 33.0 9.1 March 3. March 15. March 29. April 14. a rs) 38.5 19.5 $ 43.7 8 60.2 34.5 9 67.0 b 8 39.0 21.8 9 44.7 10 62.0 46.5 10 92:7 fe 9 36.5 19.0 10 45.0 10 59.8 40.5 10 62.5 a x! ——oe On the Physiological Influence of Manganese Compounds on Plants. 183 The ears developed in the following number: May I. May 3. May 6. May 7. a 2 6 6 7 b 3 6 6 7 O O O On May 6 a photograph was taken (see Plate XVI). The leaves of the plants (a) were paler than those of (b) and (c) and many of the lower leaves turned brownish! what was less the case with (b) and not at all with (c). Also the roots of the manganese plants turned gradually brown.? The solutions were renewed on Febr. 3, March 3, 15, 29, April 14 and 28, increasing the amount of MnSO, from March 3 to April 14 to 0.004%. Since now a parasitic fungus appeared on the leaves, the experiment was terminated on May 14. The final observations were as follows: Number Fresh Total weight| Weight Weight Weight Length, of weight of of of of ot ears, ears, dried straw. | living leaves. | dead leaves. | dried roots. cm. grm. grm, grm, grm. | grm, a 59.08 7 2.4 5.6 3.2 as 1.2 b 59.09 8 3.5 8.0 7.0 1.0 3.0 c 48.48 4 1.0 7.4. 6.5 0.9 2.2 The stimulant effect of manganese on the wheat plant becomes therefore very evident, when the plants (b) are compared with the plants (c). The plants (a) suffered evidently from the want of sufficient amount of iron, which was more evident in the later stage of development. It deserves to be pointed out especially, that the wheat plant can overcome the injurious effect of manganese much more readily than the so closely related barley plant. 1 The brown points did not here appear as distinctly as in the case with barley. 2 In comparing (A) with (B) and (a) with (b), it appears that the increase of iron had a counteract- ing effect upon the influence of manganese, not only in regard to the yellowing of the leaves but also in regard to the stimulating effect, produced by the manganese salts. This same inference can be drawn from the observations made on the radish shoots (see above p. 178). The brown color of the roots was due to some adhering MnQ,. Experiment with Pea. On March, 8, shoots of pea (3-4 cm. long) germinated in saw dust were placed in the following solutions and observed during the first period of development, that is, until the mineral and organic nutrients of the cotyledons were consumed. The plants were kept in a warmroom. Of mineral salts, ferrous and manganous sulphates only were applied. a. 0.0029 MnSO, b. 0.002% MnSO,-+0.002% FeSO, Gi 0.002% FeSO, The following observations show the growth of the shoots: Length Fresh weight. March 22, | March 27. | March 31. April 7. April 20. cm, cm cm, cm cm orm 2d 15.0 18.0 23.0 30.0 39.2 6 b 11.6 14.0 16.5 21n5 30.5 1.2 € 11.6 13.8 17.5 23.0 33.0 elt The accompanying photograph (see Plate XVII) was taken on March 31. It deserves mentioning that the yellowing of the leaves, observed with radish and barley, did here not make its appearance in this first state of development, which may be explained by the presence of a sufficient amount of iron in the reserve stores. On the Physiological Influence of Manganese Compounds on Plants. 18s Conclusions. Manganese salts exert on the one hand an injurious action and on the other a stimulant influence on plants ; with increased dilution the former diminishes while the latter increases. Thus a dilution can be reached in which only the favorable action of manganese becomes obvious. Manganous sulphate added in a dilution of 0.002% to culture solutions! exerted a stimulant action upon radish, barley, wheat, and pea. Iron seems to counteract to a certain degree the action of manganese. The intensity of the color reactions of the oxidizing enzyms of the manganese plants exceeds that of the control-plants. 1 This was of course transformed into phosphate in the culture solution. t ’ . ve of ye i \ ; e : * ‘ , ; - - . ‘- as <4 , . i ’ ; x , Ea) + § | ae 4 3 = * ‘ - te - - ’ is é ' i - © on ace ¢ a v i a= ee it 7 es | : ad aia bys °oL1 odud OF, - JOOYS YSippva UO Osouvsuvut Jo oou INYPUL OLY AULWOL Wd walle } FF aa bd ah 0 ; patil A a vel me 7 ) : | a i o” im, “y° | va ATX HLV Td ZL LOMA. LLOD ica) ILL: Wh) JOOY AQP AUC lo PSIUVOULUE tO WUT AX ALFTd wl LOA TZO). DINODF~ 77 ng VEN MeN hi SHENK CS (HOMIES WA OV by VA EL AD ES AOL SRT i= > AUILML, SMEIKMCS (LOVE) 6, WOVE, Wa. LIAL EE, SVL Plate showing the influence of manganese on pea shoots, To page 18 On the Action of Sodium Fluorid upon Plant Life. BG K. Aso. With Plates X VITI—XIX. Although it is very well known that sodium fluorid exerts a poisonous action on plants and animals, the writer has made some further experi- ments with the view of determining the point of dilution at which that poisonous action disappears and a stimulating action sets in. Ono has inthis regard observed that sodium fluorid in a dilution of 0.000.03% has a stimulating action on the development of alga. Experiment with Seeds. Seeds of rice, wheat and mustard (50 of each kind) were steeped in solutions of 1%, 0.5%, 0.25%, 0.1% and 0.059 NaF for 48 hours and then, superficially dried, left to germination in purified sand. The number of germinated seeds were counted after 6 days with the following result : NaF, Wheat. Mustard, 1 % I oO 0.5 % 2 oO 0.25% 2 oO oO. % 2 6 sf 0.05 % 7 14 Control, 16 23 It will be noticed that the rice seed had more resistance power toward the poison than that of the wheat and mustard, what is probably due to 1 Journ, College of Science ; Imp. Univ., Tokyo, Vol, XIII. part I, (1900), With fungi, a stimu- lating action was observed by him in the concentration of 0.005% NaF, Yeast is injured by 0.019% NaF. 188 K. Aso: the thicker cover, preventing the ingress of so much poison as entered in the other cases. The shoots were largest in the control case but these were closely followed by the shoots from the seeds treated with the 0.05% NaF solution. Very striking, however, was the rapid falling offin length between these and the shoots from the seeds treated with the 0.19% and the stronger solutions. Ina second trial, seeds of soy-bean were compared in regard to the resistance power with seeds of wheat. They were steeped in solutions of NaF of the following concentrations : 0.05 % O1On" 5 0.005 ,, 0.001 ,, After 24 hours, the solutions were poured off and the seeds left to germination, 20in each case. After g days, the following data were observed : Number of germinated sceds. Average length of plants. Sodium fluorid, Fe Wheat. Soy-bean, Wheat. Soy-bean. plumule. radicle. 0.05 % 7.ccm. 8.2cm. 3.0cm, Gor 4 FO. 9.6 5, ©:5irs: 0.005 ,, Si5) ss TO SOs, 0.001 ,, 8.5/5 TO%,s 6.5 5 Control ,, HeOnss 10.5 95 9.0 4 Also in this case, therefore, like in the first, sodium fluorid in the cons centration of c.959§ acted injuriously upon the seeds. Experiment with Soy-bean Shoots. Shoots of soy-been were (March 18) placed in solutions of sodium fluorid of the following concentrations : a? On the Action of Sodium Fluorid upon Plant Life. 189 NaF. Length of shoots. L OI % Io Cm, b C.05 , tomes c 0.01 i) >. d 0.005 ,; IO » c Control ,, | 9 After three weeks, the shoots (a) and (b) had withered while with (c) and (d), the leaves remained green and perfectly healthy and the height of the shoots did not essentially differ from that of the control case. The cotyledons (c) and (d), however, had lost their green color and their turgor, and had become yellow. A slight touch sufficed to cause their dropping off. This indicates that the poison was retained to such an extent in the cotyledons that only traces reached the leaves above. Experiment with Pea. On March 6, 1902, pea shoots 2 cm. long were placed in solutions of 0.01, 0.001 and 0.0001% sodium fluorid. The results are shown by the following,table : | Length. a Fresh w eight dete on April 22 March 31. April 14. April 22. 0.01 % 17,0cm, 20.4 cn), 20.4 cm, r.2 grm. 0.001 ,, 19:0 .;, 26.5 32.5 1.3 0.0001 ,, 30.3 > 36.5 I.4 Control. 29.0 , Ly fe ee I.4 A poisonous action on fea shoots is therefore produced by 0.01 and 0.0019§ NaF, but it is not any more noticeable when the dilution reaches 0.0001 9%. 190 K. Aso: Experiments with Barley Shoots. I. Shoots of barley were placed (March 25) in a culture solution to which was added NaF in the following proportions : a 0.05 % b COO lem Bi, c 1005s: d O.OBT 5, e€ Control case. The result is seen from the following table : Length. Number of stalks, April 4. April 8. April 16. April 25. April 8. I I I pri 25 P April 16, March 25. | — | | a 29.3 cm. 29.2 cm. 28.2 cm. 27.0cm, 4 4 b 37-5» 37-2» 37-5 40.0 ,, 4 4 c HOO) BO) gp. 40.0 ,, SEOs op 4 4 d 28.2 5, 30.2, 33-5 435 » 4 7 35-5 99 On March 30, the leaves of (a) lost their turgor while with (b) the tips of the leaves became yellowish. On April 1, the lower leaves (a) had died, while with (b) they were injured and with (c) they showed yellow tips. No new rootlets had appeared with (a), while a few with (b) and more with (c) and (d). On April 4, the general appearance of shoot (d) was perfectly normal. In this case, therefore, a stimulating effect of sodium fluorid in regard to the increase of the number of stalks of barley at a dilution of 0.005 and 0.001% was quite evident. II. On October 15, 1901, barley shoots 1o—12 cm. long were placed in a culture solution to which were added : 1 The measurement relates only to that part that still was healthy. On the Action of Sodium Fluorid upon Plant Life. 191 a 0.01 % Sodium fluorid. b CHOOT «<2 Pe 0.0001 ,, " Control. The following table shows the observations made Length, Number of stalks, J rst Wei, Fresh Weight Octaar. | Dec. 12. | Febr. 6. DEG, 12% | Febr., 6. oe Febr. 6. a 15.3¢m, 26.1 cm. 27.0cm, 5 9 "agli TY b 1G;015 nekciere Sy Melee 7 10 c 19.5» 39.0 ;; 5 = d E735 = 35 34. 35 5 7 The plant in (c) which developed most was injured by the severe cold in the winter. Very many root-hairs and rootlets appeared in (c), also they developed very well in (b), but poorly in (a). The remarkable effect of sodium fluorid in very high dilution upon an increase of the stalks was also here observed like in the former case with barley as the table shows. The accompanying photograph (Plate XVIII) was taken on Dec. 12. During this experiment, the solutions were renewed on Nov. 5, 25, and Dec. 17. Experiment with Wheat Shoots. On May 18, shoots of wheat 6.5—7 cm. long, were placed (3 in each case) in solutions of 0.001% NaF (a), and 0.oo01% NaF (b) to which the necessary mineial nutrients were added. Towards the end of May, it became evident that the growth of the plants (a) was much slower than that of (b) and of the control plants. On June 12, the following data were observed: 192 K. Aso; Average number of leaves Average length of one Total fresh weight of one shoot, shoot, of 3 shoots, a 3 PTA ‘Cit, 0.8 grm. b 4 204 ~,, URS 5, Cc 4 2Oe1...55 0 see A poisonous action of sodium fluorid in the dilution of 0.001% was therefore quite evident, while in the 10 times higher dilution the poisonous character was not observable. Since a fungus commenced now to show on the leaves this experiment had to be terminated. Experiment with Rice. I, Shoots of rice 8—1ocm. long were placed in solutions of 0.01% and 0.05% of NaF, but in the latter solution no development took place and the tips of the leaves turned yellow after a few days. In the 0.01% solution, some growth was observed, but less than in the control case, demonstrating the injurious influence at the dilution of 0.01% upon rice. II. On June 12, shoots of rice 22—25 cm. long were placed im the same solutions as the wheat shoots (2 shoots in each flask). After five days, some difference of development was noticed, especially with the roots. After 10 days, the following data were observed : Total number of ‘Total number Length. new rootlets, of leaves. a. 0.001 % NaF 32 10 32.0 CM. De (0.0001... a7. 8 30:4. 55 c. *Contiol: 20 6 33:00 ys A certain stimulant action is therefore noticeable in the case (a) and it appears therefore that rice is not so easily injured by NaF as wheat. Experiment with Flower Buds. On Febr. 4, three plum branches bearing 5, 7 and 10 buds respectively were placed in solutions of Nal of various concentration. The following data show the result: =— BP Mb bew oro Gus wy On the Action of Sodinm Fluorid upon Plant Life. IC Ww Number of flowers developed. Febr. 24, Febr, 25. Febr, 27. a. 0.01 9% Sodium fluorid. O O fe) Be O00. , sg 3 4 6 Ex O00! ,, As a 3 3 d. Control. 5 O 2 3 There had no bud opened in the solution of 0.019% NaF, which shows a poisonous influence in this concentration. Very striking was the different size of the flowers; the petals in the control case were much larger than those in (b) while those in (c) had an intermediate size. Hence we observe on the one side a stimulating action of sodium fluorid as to the time of development of the flowers, but on the other, a diminishing effect on the size of the petals, a notable parallelism to the. case of the barley shoots (see above). Experiment with Leaf Buds. Branches of Cornus macrophylla JVal/, of equal length and size bearing 4—5 winter buds were placed (March 4) in solutions of sodium fluorid of the same concentrations as just mentioned. The result after 37 days was as follows: Original Number of buds number of buds. opened. Remarks, — eee March 26. March 27, April ro. a. 0.01 % Sodium fluorid 5 O fe) I sa mae COOL ds 4 4 4 4 =. O.000! ,, . 5 4 5 5 d. Control. 4 3 3 4 The size of leaves was largest in (c), smaller in (d), and smallest in (a) and (b). Here also a certain accelerating influence of sodium fluorid in very high dilution upon the development of leaf-buds was evident.! t It may be mentioned here that also some experiments were made with injections of a 0.2% solution of sodium fluorid into young branches and buds, but besides the death of the tissue around the point of injection no striking effect was noticed. 1904 K. Aso: Experiment with Pea in Soil Culture. Two pots each holding 2.3 k. soil served here for the experiments with peas, of which 15 seeds were sownon Febr. 19. The young plants were reduced however, on March 27, to five equally large ones in each case. Each pot had received the following manures : 4.6 erm. Common superphosphate. 250" 5, Potassium carbonate. B10 sds Sodium nitrate. While the plants in one pot were treated with sodium fluorid, the other pot served for control. The pots were kept in the green-house almost all the time. The amount of sodium fluorid supplied on each application was only 0.001 grm., dissolved in 100 c.c. water. The days of application were as follows: March 11, 25, April 14, 21, 28 and May 6. The total amount of sodium fluorid was therefore 0.006 grm. and still in spite of this small quantity, a stimulating effect was gradually noticed and was finally also recognized by the weight of the seeds produced. The formation of flowers commenced on April 22, and was ended on the 16th of May. One day afterwards a photograph was taken (see Piate XIX), which shows that under the influence of sodium fluorid the plant had reached a greater height than the control plants.! Up to the flowering period, almost every day 300 c.c. water for irrigation was applied, later on the quantity was increased to 500 C.c. The fruits had ripened on the 2nd of June, and were weighed in the fresh state, while the straw was weighed in the air dry state. The results wer2 as follows: 4 On April 23, some plant-lice made the appearance on ihe leaves and from now careful search was kept up and every louse noticed killed by touching the insects with a little brush moistened with a 196 carbolic acid solution, On the Action of Sodinm Finorid upon Plant Life. 105 Sodium fluorid, Contro Werohtoftreshi fruits, .....0.:-...->. 71.7 grm. 60.5 grm. Weight of air dry seeds ............ BZN 35 23.2 WWEIGME OL SUVA Witenes «cs canedaces+e oe Wess 10.7 This result doubtless shows that a stimulating action by this small quantity of fluorid had taken place. —_—— — <3> - — - C- ° § F etioeentin § s ; ‘ ; - ' _— . oan pe ba a /_ ; j . . - t 4 3 a yw y oo“ . ; > : . ° — — é 1O1 onud OT, *AgpAvq UO SUOIVAQUNUOD JUMOYIpP UL Pony UINIpOs Jo ddan YUL 94) SULMOYS oyLI{ 4 Ny TMAX PIVITd LOM AIO) DI LIE SL ' ? ! ’ ‘ t IC Mell, NE KM, (OYE, VWAQVG, VW PEATE. GON: iy I. Plate showing the influence of sodium tiuorid on pea. T Fluorine > IT control plant. To page ro4. \ ~ . On the Action of Sodium Silico-fluorid upon Plants. K. Aso. The foregoing observation with sodium fluorid made it very probable, that sodium silico-fluorid would also prove a poison even in a considerable dilution. In order to observe however whether this salt would in still higher dilution exert a stimulant action, the following experiments were made with shoots of soy-bean and barley (about 25 cm. long). To the culture solutions were added: a. 0.05 % Sodium silico-fluorid. b. C.01 ,: ¢ ag G. 0,005 ,, Pe ‘3 d. 0.00I-,, * ¥ e. Control The observations were as follows: — “wee _oq_cswo_1€é_e—oxJycewm i cq Soy-bean. Barley. March 27, Shoots (a) and (b) lost turgor, ae Shoots (c) lost turgor, The tips had all turned yellow with the shoots (b) and (c), while with (a), all leaves had withered, Leaves with (d) still normal. 30 Shoots (d) stationary. Considerable E * | development in the control case, Almost all leaves with (b) and (c) are dead; only April 1, Shoot st turgor : avy Gee pril i root (d) lost turgor and withered, shoot (d) and the control shoot were still healthy, Control shoot normal, has grown 1o cm, shoot (d) 8. Only control s still alive oD 1 . oe nly control shoot still ative, still alive, but stationary, RE 198 K, Aso: On the Action of Sodium Silico-fluorid upon Plants. It will be noticed that sodium silico-fluorid is a strongerspoison than sodium fluorid when the result of the foregoing article is compared with these. In the solution containing even only 0.005% sodium silico-fluorid, the barley as well as soy-bean shoots had been killed almost completely in 6 days. Buta remarkable effect was noticed with barley shoots (d), i.e. where the dilution of sodium silico-fluorid was 0.001%. While here growth in hight was very sluggish there had developed up to the 8th of April from the originally three stalks as many as seven new stalks, while not a single new one had started in the control case ! This forms a third instance of this kind of action of a fluorine compound, since sodium fluorid produced the same phenomenon with barley shoots in the two cases described in the foregoing article. It deserves notice, further, that soy-bean is more easily injured by highly diluted sodium fluorid and silico-fluorid than barley. — ——_. f | F | The first reaction (a) was oe ao + + ae ae + stronger and the fifth (f), y: weaker than the rest. Germinated ; | wheat. * 3 "i | Si a | ii Germinated | rice, oF + + | + B |. o-oo | (OL ee Germinated soy-bean, + + = 8 23 P The violet color with ; Aspergillus (f) appeared along the eee oF 3 rim, where the fungus boiled rice. : _ The plumules of the germinated plants were about 1 cm. long and the tests obtained showed the distribution of the oxiding enzyms to the whole Ls extent of the sections. The tests obtained leave no doubt that the oxidase which gives a blue reaction with guaiac tincture (laccase?) increases during germination while spermase decreases, which is in accordance with the observations of Griiss, that the reaction for spermase which at first increases, decreases later on, until it fails altogether. | The second series of tests were made with various vegetable juices. The objects were crushed in a mortar with addition of some quartz sand and extracted with some water. The filtrates behaved as follows: K. Aso: 216 “ec “IO]OI an{q v sonpoad 0} JUSIOYJNS Jou sie aiNyOUy oeIeNS jo sdorp Moy v ‘(v) YIM Suyso} UT *ASUDJUT JII.M SUOIIVII ]PV “ ‘Quoye (2) asvq oy} YA UOT ON *ISUDJUT .1O.M SUOTIVAL [TV ‘poarvadde 10j09 JQJOIA B QuETR (9) aseq OY} UTA "SALUT -uoAuoydeaed -uofAuaydeared -Ayjouresja J, *[eaynou SOOUY) ‘prow Aqure sy *yerynou soupy “poe Apysys £19, “ouOTL yoo aarnyouy ovieny | aanjou ovieny| jo wove “q oe *s}ooys-ooquirg “PIE JO yma pousdny "ysIpey Jo 00Y “vyLeq Jo JOoyy “O}L}OI jo som 217 the Vegetable Body. Enzyms in idizing On Ox “paso, spalqo aquyadaa [jv ul judsoid sea asepeyes » *S}S9} OSOY} LOJ POAJOS “Oye 9[VP V UL paayossip ‘oyrydioaad oy. pure “oyoore yA payeydioaad sea yovsyxa snoanbe AL e *s]S9} Of} AOJ PAAAIS “DWM IAI] V UL POAOssip ‘aqeyidioaad oyoyoore ay} doUdzY “suONOLVaA dso. [[C YWA\ soadptoyUy UFUUL} OUTS ‘asuye;vo Joy ylaoxo uoNOveA Auv aad you pip oof ysaay ayy, z “auoye (9) aseq oy} WEA porvodde uoyovas yYysYS *Ayuenb oyesropout ut asvypeyey "UONOLVOA DSUDJUT OAVS asryLyeD “AyQuenb oyerspour ul yasvpeyeo “O9SUD}UL AIO.M SUOTPIVOA [PW ayeydysoad gayjoyoore ayy UEAK ‘sadud snuryiy yy poysay sea uOnOvaL OY, 4 + + + + + px seen soupy 990.1) = 99v.] D9v.y = § d9vy iG « + be ate + fe av ae “ | ats + ; F ot ae 3 cen ‘pre Apurey HEWN 2 oy “ysvadk-1994 “Ayeyy wtoay o99%q0} paan,y *“o99Rq 0} asourdef paing (“ysaay) “oo0B qo], Jo SOATIT "BAT, JO SAATIT 218 K. Aso: Also an animal secretion, saliva, was compared in regard to these tests with the following results. The samples came from different persons. | ; a. b. c. d. e, 2 guaiac A eect actin tage tetramethyl- guaiac nctare. guaiacol. | paraphenylendiamlne. paraphenylendiamine tincture, +H,0, +H,0O, +H,0O, +H,0O, ie _ trace, + trace. trace. I - - ae fo) fe) | ioe] = — 4 fe) fe) IV trace. -- 4 = fe) | | V. trace. a + itEace: trace. While the guaiacol reaction (c) was intense in every instance, the fourth (d) and the fifth reaction (e) appeared merely in traces. The guatac reaction for oxidase and peroxidase was obtained only exceptionally in certain samples of saliva. An aqueous extract of cow’s liver and horse kidney did not yield the guaiacol reaction, while extract of cow’s pancreas gave it, although weaker than saliva. The pancreas also yielded a blue reaction with guaiac tincture and hydrogen peroxid, while liver and kidney in this case failed to produce it. Exist there several peroxidases ? Although I was unable to find any vegetable object which would give the guaiacol reaction in absence of peroxidase, the behavior of saliva shows clearly that the guaiacol reaction is—at least in this case— not due to the common peroxidase, recognized by the blue reaction with guaiac resin and hydrogen peroxid. We have therefore to distinguish two 1 Control tests were made with boiled saliva, and in no case a coloration sets in. On Oxidizing Enzyms in the Vegetable Body. 219 kinds of peroxidases, one that gives the red reaction with guaiacol and hydrogen peroxid, the other that gives a blue with guaiac 1esin and hydrogen peroxid. It seems further probable that there exists a third peroxidase! which gives a violet coloration with tetramethyl parapheny- lendiamine in the presence of hydrogen peroxid ; the reactions mentioned in the above tables are in favor of this view. Though a certain parallelism between the intensities of these color reactions was observed in some cases, it was not recognized in others, hence the view that each reaction might be caused by a separate enzym, is justified. On the other hand, one might object that while there exists a separate peroxidase that gives the red guaiacol test in saliva, there is no proof that the common peroxidase does not give both reactions at the same time, since thus far no object was observed that would give the blue guaiac—H,O, test without giving also the red guaiacol test. In order to test this objection I have made the following comparisons as to the killing temperature of oxidizing enzyms. The fact that the acidity of the plant juice, the degree of dilution, the duration of heating and the presence of certain salts have a modifying influence on the height of the temperature at which the change of an enzym to the inactive modification takes place, was pointed out by Loew? in his ‘ Physiological Studies of Connecticut Leaf Tobacco.’ Since those influences have not always been paid careful attention to, the existing discrepancies between the data obtained can hardly surmise us. The following table shows the various data obtained thus far : 1 It seems to me very probable that the reaction with paraphenylendiamine and hydrogen peroxide, and that with tetramethylparaphenlendiamine and hydrogen peroxid might be caused by the same enzym, 2 Report of U.S. Department of Agric., No. 65. 1900. p. 21. Name of the Temperatur which kills the Duration of Remarks, Name of the author, aN ating. enzym, enzym. Heating An oxidase in a ; . Qe , —— os Urushi, 63 iC Voshida. An oxidase in an A 5: above 70°C a ——— Bertrand, Urushi (Laccase). hear . a c ears Injured at Tyrosinase, below 70°C an 50°C. % Oenoxidase. rae. 4 min. a Martinana. 33 55cC. 1} hours, ——_ » ‘ Oxidase in the stalks of the 60°C, oe —— Raciborski. sugar cane, ie F I part dry leaf Oxidase in a tobacco an : Bak tae 66—67°C, 3 min. was exiracted Loew. leaf, 9 . with 2oparts H,O - é = . f ion - Oxidase in tea-leaves. 76—77°C. 5 min. The solutior . The writer. : was neutral. Peroxidase in the stalks = P ‘ dase in the stalk 95°C. SS Raciborski, of the sugar cane. Peroxidase i E ors P eroxidase in tobacco 87°C. 3 min, leaf. : The reaction of Lover, Peroxidase in 33% val : : 99/0 or. a few seconds, sol alcohol solution, 7 the solution - was neutral, 5 Peroxidase in 10% 20° after a short / 93 . ti ime, (NH,), SO, On Oxidizing Enzyms in the Vegetable Body. to to _ = Temperature 3 4 = Name of the . which kills the ee re Si Remarks, fear? of the _ enzym, énzym: eating. author, oe cocoa: on boiling. Still active. Hunger. Salive oxidase which P produces guaiacol g2°C — - = ie Dupouy. reaction, : B-catalase in a tobacco 2 more than us leaf 72°C. 15 min, Loeu is ACG: very soon, ——— = The greater part a 75°C. iar ade caiered. a -a-Catalase in a tobacco : Faint trace of the leaf, 75°C. Soe reaction, ss se I min, An oxidase in milk which gives Storch’s reaction, In order to decide whether the reactions caused by hydrogen peroxid with guaiac resin, guaiacol, paraphenylendiamine, and tetramethylpara- phenylendiamine disappear at one and the same degree of temperature, the following tests were carried out. Potato-, bambooshoot-, barley leaves-, and radish root juice were heated for a short time to 80°C or over with the following results : (+) This forms a noticeable exception from the general rule that enzyms are killed below the boiling heat of water. According to Zinossier peroxidase of pus is not destroyed at 120° when dissolved in a weak acid medium, (La semaine med. vol, 18), A similar observation was made by Spifszer with peroxidase from animal organs (Pfliig, Arch, vol. 67, p. 615). N N “Oy R.19 pout *O]R.19pout *prxorod uasorpAy pure aumueipuaAuaydeard] Ayjouresj9} YA JY} UO vor yoJOIA OY} £ prxoszod udZo01pAy pue oururipusjfuoydeaed yy yey Uoyover ud913 ay} { prxosrod ussorpA ture Ipuos] I [AY 1}! l JO!A 9U} + Pl pay Pp TwWeIpus]AUdy UH |} Uo! Ui + pl pAt pur Joorrens yy uorovoas ay} suvoUT UOIPLAI pat oY} sNYy} ! poonpord AojOd dy} 0} Surpxogdv_paourvu ov SUOTPLOI dSdY} OPLIADIGde O} JopsO UT + “SUISvOIOUL Ayenpras *Q}v.19 pout ynq ‘yvom *O}VINpOU “DUO elual *3u0.a4s “QUOU Paya royet *SuoT4s “QuOU “uU0U *8u0.19S “ou0U bau *3u0.1]s “.U0U “QUOU “Q9vA} WSS | “oov17 WSs "url £ “UN S “ur € “088 ‘Dof8—z8 ‘08 ‘uoyores prov Ayysys AroA ‘oor ysrpea Jo son fi —— ae “ou0u *ouOU Palnel *“Suo0r4s "20RI} “DOVAP JULSIYS ‘ouou ‘Je8S-=28 “plow yrom ‘saavoy -Agjaeq Suno¥ jo oon [ "uy § "UM $ “9.08 “088 *jeajnou jsouye *ploe van ‘sjooys-ooquieq jo soinf ‘goint 072}0q i a q UOT UdeIT) ‘ISLPIXOIIT jo uojjeinp pue a0159(T On Oxidizing Enzyms in the Vegetable Body. 223 The influence of concentration upon the killing temperature was observed with the enzyms of cured tobacco-leaves.1 A leaf of cured tobacco (5 g.) was extracted with 100 c.c. water, and tested with the following results : Degree and 80°C, £4°C, 85°C, 86—87°C, 90°C, duration of heating, oF 5 Min 5 Min. 5 Min. 5 Min. 5 Min. ee Oxidase. trace, none, rone, none, none. Peroxidase. strong, weaker, weaker, weaker, trace, Green reaction. strong, weaker, weaker, © weaker. none. Violet reaction. strong. weaker. weaker, weaker. none, Red reaction. strong, weaker, weaker, weaker. trace. A portion of that extract was diluted with an equal volume of water (a), and another with twice its volume of water (b). Upon heating to 87°C for 2 minutes, the following result was obtained : Original solution. Oxidase. none, none, Peroxidase. trace. none. Green reaction, none, none, none, Wiolet reaction, none, none, ‘ none, Red reaction, strong, moderate. 3 trace. Though the killing temperature is not a constant magnitude under all conditions, neverthless these comparisons show very clearly that the red reaction caused by guaiacol and hydrogen peroxid is due to a separate 2 The reaction of the extract of the cured leaves was neutral. 224 K, Aso: enzym which has more resistance-power towards heat than the other related oxidizing enzyms. The enzyms which cause the green and the violet reactions have an intermediate resistance-power between that of oxidase and that Of peroxidase. That there exist also quite a number of different oxidizing enzyms acting in absence of hydrogen peroxid, can be inferred from observations of Bertrand and of Bourquelot. The latter has observed among other things that when the extract of the fungus Russula delica was kept with some chloroform, at first will be lost the oxidizing action upon tyrosin, later that upon guaiacol, and finally after eight weeks also that upon guaiac.! Influence of Foreign Substances upon the Color Reactions. The investigations on the behavior of oxidizing enzyms towards foreign substances are very incomplete. The power of the oxidizing enzyms of an animal organ is not injured by certain poisons or by freezing. Only hydrocyanic acid and hydroxylamine hinder the action. 80% alcohol does not kill oxidizing enzyms, but a stronger one will injure them slowly. Acids and alkalies act injuriously. While chloroform may promote the action of certain oxidizing enzyms,a prolonged contact will act injuri- ously. Of some interest is the comparison of the behavior of catalase? with that of other oxidizing enzyms : Salts of a strong acid or alkaline reaction injure this enzym while salts of a neutral reaction do not. A remarkable fact is the depression of the activity of the soluble or -catalase by nitrates, while the exzym itself is not injured by them. In general, the retarding influence of a salt is increased with the amount of the salt. Sodium carbonate attacks B-catalase slowly ; 29¢ sodium fluorid and 59% dipotassium oxalate solutions cause no injury. 5 potassium sulphocyanid and thiourea interfere with the catalysing action, but have no direct injurious influence upon the enzym. Mercuric chlorid acts very injuriously. While highly dilute acids 1 Bourquelol: J. B. f. Thierchem, 26. p. 886. Cf, also his observations on the potato (ibid). 2 Joew: Catalase, an enzym of general occurence. U.S. Dept. of Agric, Report No. 86. On Oxidizing Enzyms in the Vegetable Body. NO iS) vi retard the action of catalase, dilute alkaline solutions promote it. When the amount of a mineral acid in the solution reaches more than 0.5%, catalase is soon killed. At the ordinary temperature, 0.19§ acetic acid does not injure Catalase in one hour, but gradually at 55°C. 2% sulphuric acid destroys the power of catalase in fifteen minutes. Saturated baryta solution injures a- catalase slowly, and kills -catalase intwo days. While 0.19% caustic soda Causes no injury, 196 of it kills catalase instantly. Alcohol above 309% has a retarding influence while more dilute alcohol or absolute alcohol does not injure catalase within twenty hours. Little chloroform and ether have no direct influence on the action of catalase: 19 phenol retards the action of the enzym; 5% formaldehyde destroys its action in a very short time; 0.4% nitrous acid injures the enzym considerably in one day. a-catalase shows considerable resistance to the action of hydrocyanic acid of 2%, while #-catalase is gradually killed by it. Afier the evaporation of hydrocyanic acid, regeneration of the action of 8-catalase is only observed, when the amount of this acid has been very small. Hydrogen sulfid and also phenylhydrazine injure the insoluble or a-catalase but slowly at the ordinary temperature, also a 5% hydroxylamine solution does so. In view of the interest connected with the chemical behavior of enzyms I undertook a series of analogous experiments with the oxidizing enzyms mentioned above. My observations were as follows. 1. JLnufluence of salts upon the color reactions mentioned. 5 c.c. of a dilute juicet of radish root were mixed with various salts and left with some drops of ether for 48 hours before testing. In carrying out the oxidase reaction I c.c. of guaiac tincture was added ; for the green reaction, I drop of sodium acetite, 1 drop of para- phenylendiamine and 2 drops of hydrogen peroxid; for the red reaction, an equal volume of guaiacol solution and 2 drops of hydrogen peronid ; for the violet reaction, 2 drops of an alcoholic solution of 1°/.) tetramethyl para- 2 It is necessary to dilute the juice moderately to obtain the color reaction of a degree saitable * for comparison, 226 phenylendiamine and intensity of the color produced was compared with that of the control solution without salts. K. Aso: drops of hydrogen peroxid. In each case the Salt. Conceniration. | Oxydase. Red reaction. | Green reaction. Sodium chlorid. 5% weaker. weaker, weaker, Potassium e ees Bn weaker, weaker, weaker, nitrate, a Magnesium slightly avid > O7 ous y oir = rong strong nitrate. 570 weaker. gay, ens. Calcium a nitrate. 570 2? . 9 Magnesium Pia ae ce Or strong sir strong sulphate, 5% strong. strong. strong. Ammonium oe sulphate. 5A 33 ”? > Sodium : 0 sulphate. 5% 2? 3 » : : io! Dipotassium ay aaVer a.wiolet color phosphate, We weaker. weaker, appeared i instantly. Monopotassium iy fea phosphate, 570 strong. strong. strong. Sodium t 1 quickl = 5 0/ PRE i A Aas urned quickly to carbonate. “70 weaker. weaker, violet, Potassium ov een reaile not green, but oxalate. 1% Caker, weaker, violet, Ammonium Ov. at : ' ' y TOU sti op we Secale 2% strong, strong. strong Sodium ati . moderately moderately fudsid 2% none = : u : strong. strong. Sodium by ine nayaaee ; silicofluorid, O37 hia ea ae —— ne ow a Violet reaction. weaker, strong, strong. 3”? 2? ” 9 weaker. strong, slightly weaker. almost none. weaker, moderately strong. none, On Oxidizing Enzyms in the Vegetable Body. 227 It will be seen that sodium chlorid, potassium nitrate, calcium nitrate, magnesium nitrate and dipotassium phosphate in a concentration of 5% injure these enzyms, or weaken at least the color reactions caused by them; of special interest is here also the influence of potassium nitrate, since this depresses the action also of catalase, without injuring the enzym itself.+ Sodium carbonate (2%) and potassium oxalate (1%) weaken also these color reactions. The influence of sodium chlorid (5%) and potassium nitrate (5%) was tested once more, this time after 24 hours, with a more concentrated juice of radish and also with milk, and a decisive depression observed, especially in the case of chlorid of sodium and in regard to the red reaction. The striking influence of sodium fluorid and of sodium silicofluorid on the appearance of the color reactions led me, not only to repeat the tests with radish juice, but to make also some further tests ; 3.5 grms. of air dry tobacco leaves were crushed in a mortar and extracted with 300 c.c. water. The filtrate served for the following tests : Concen- Red Green Violet - : $ = Oxydase. | Peroxydase. : : a Time of testing tration, ) ) raection. reaction, reaction. S Sore a ee J SS Sodium - fluorid. trace, weaker. weaker. }| weaker. | weaker. Immediately. After about one hour. none, weaker, weaker, weaker, weaker, Immediately, weaker weaker | weaker Sodi weaker than — : yea jum dene ae een than in than in than in silicofluorid, . of NaF. the case the case the case Pair of Nak. of NaF. of NaF. | After about none. . 2 ” ’ ’ ’ one hour. ce ee a ce Re none, ‘ ‘ . Immediately. K. Aso: In the next test, 50 grms. of malt were crushed and extracted with 300 c.c. water and a similar effect observed with this filtrate. Oxydase. Peroxydase. Time of testing. Sodium fluorid 5% none, weaker than control, tested, immediately. Sodium silicofluorid 0.3% none, slight, It follows therefore that sodium fluorid and _ silicofluorid have a injurious influence on oxidizing enzyms, especially on oxidase proper. The sodium silicofluorid acts fuither more energetically than the fluorid. 2. ILnfluence of dilute acids and alkaltes on the color reactions. The juice of radish root rendered slightly alkaline with caustic soda, yielded no blue reaction for oxidase or peroxidase. The greenish colora- tion produced appeared also in the absence of the enzyms. The red reaction set in, but slightiy, while the green and violet reactions failed to appear. Upon acidifying these solutions, however, with acetic acid, the oxidase and peroxydase reactions as well as the other reactions mentioned appeared with great intensity. Moreover, when these solutions were made again weak alkaline with caustic soda, these colors disappeared again, and reappeared on adding some acetic acid. 100 germs. of a fresh radish root were crushed and extracted with 300 c.c. water. The filtrate served for the following tests, which always were made after neutralization of the juice. 1 Other enzyms seem to have more resistance power toward sodium fluorid, Cf. 47/Aws and Lhiber, Jahresb, Thierchem, 1893, p. 640. NN On Oxidizing Enzyms in the Vegetable Body. 229 Acids or Concen- Time Red Green | Violet : : ae Oxydase, | Peroxydase, a Se : alkalies, tration, | after mixing.| ~*~) a YeAse- | yeaction, | reaction, reaction. ; | ee drochlori | , Hy a ‘a c 5 hours, none, none. trace, trace. | trace. See SS ef d%a See Nitric : acid ” none, none, none, none, none, _ Sulphuric = acid, ” ” ” 2 7 | ” *- Acetic acid. 3 ” ” cry ’ 2 ° Oxalic acid, Et ” ” " ” ” Caustic soda, 29 aS or) ” ” ” = aa Vl | Tartaric pate | - acid, 24 hours, ” ” : ” | ” * This was repeated with the water extract from a cured tobacco leaf with the difference that the tests were made 2— > 3 minutes after Concen- Red Green | Violet : Oxydase, | Peroxydase 2 : : tration, bi J “| reaction, reaction. | reaction, | —_-_—-—-— - -- —_——_—--_- -— ST none. trace, trace, none. none, trace. trace, weaker. trace, weaker. | weaker, strong. strong, strong. strong. ver. a none, weaker, weaker, weaker, weaker, weaker, i | : none, none, none, none, 230 Kk. Aso; 3. Lnfluence of Potsons. Influence of hydrocyanic acid: Two leaves of cured tobacco (about 10 grms.) were extracted with 200c.c. water to which 2% hydrocyanic acid was added. Hereby all the color reactions mentioned were prevented, but after removing the hydrocyanic acid by a current of air, they could be reproduced,! except in the case of oxidase. Influence of hydrogen sulphid: Radish juice and an- aqueous extract of cured tobacco were saturated with hydrogen sulphid. Upon replacing this substance by air immediately afterwards, all reactions were still obtained, But, when after standing for 48 hours the hydrogen sulphid was replaced by air, these reactions appeared very much weaker, and that upon oxidase not ot all. Behavior to phenylhydrazine: 10 grms. of cured tobacco leaves were extracted with 200 c.c. of water. To 100 c.c. of this extract, some hydrochlorid of phenylhydrazine and sodium acetate were added, whereby some yellowish flocculent precipitate was formed. After 24 hours, much absolute alcohol was added to precipitate the enzyms and separate them from phenylhydrazine, since this might have interfered with the color- reactions. The precipitate was collected in a filter, washed thoroughly with alcohol and dissolved in a little water. All color reactions failed in this case. Do Sugars Prevent the Color Reactions? It is a well known fact that tannin interferes with the guaiac blue reactions and also with the actions of myrosin and emulsin. Recently, Hlunger® observed that a reducing sugar present in the milk of the cocoa- 1 Cf, also Zfstein, Archiv. f. Hygiene, vol. 36 p. 140. Prussic acid prevents the coloration of beet juice exposed to air, while after expulsion of that acid by a current of air this oxidative process sets in again. 2 Hunger expressed in a recent article (Ber, Deutsch, Bot. Ges vol. 19, 1901), the supposition that in my tests for oxidase and peroxidase in ripened and unripe kaki fruits, it was the sugar and not the tannin which interfered with the reaction, This is however not so. On Oxidizing Enzyims in the Vegetable Body. 231 nut prevents guaiac blue reactions of oxidase and peroxidase. Moreover he observed that the more intense the guaiac-blue reaction is obtained with the sugar cane, the less sugar is present.’ 1 observed, however, that the guaiac reaction for oxidase is not interfered with by the addition of 10% glycose, fructose or cane sugar, but it requires a little increase of the guaiac tincture. Just as little as these sugars, albumin and pepton prevent the guaiac reaction. In 100c.c. of a dilute radish juice, 10 grams of egg albumin or pepton were dissolved and this mixture tested after 24 hours. The above- mentioned reactions were still obtained. © » Do zymogens of oxidizing enzyms exist? ~The ‘‘regeneration” of enzyms depends upon the presence of zymogen. The zymogens of the vegetable enzyms have been studied * but little. It is already known that there exist zymogens of pepsin, _ trypsin, rennet and diastase in the animal body. As for vegetable enzyms, zymogens of a proteolytic enzym in Vepenthes and lupin-seeds, of inulase in the resting tuber of artichoke, of lipase and rennet of the castor oil seed and of a diastasic enzym in barley and wheat have been observed.? investigated, although they must exist, as the following observations will — show. I. Juice of barley was heated to 80°C. for 5 minutes, whereupon the “oxidase reaction failed to appear, the oxidase having been changed. Was obtained. After 24 hours standing, a weak oxidase and a strong ‘peroxidase reaction were again observed. The reaction of the barley is Flinger, Het optreden der oxydasereactie in verband met de localisatie der glycose in het ‘Suikeriet, Archief voor de Tava-Suikerindustrie, 1901. : 2 Cf. RX. Green: The Soluble Ferments and Fermentation, p. 389-391. 232 K. Aso: was very faintly acid. Similar observations were made by Advert F. Woods. 2. Prof. Loew has observed that the juice of bamboo shoots which on boiling lost every trace of active enzyms, showed again a reaction for peroxidase and the red guaiacol reaction after 24 hours standing. But when, after the boiling, 19% acetic acid was added, no ‘‘ regeneration ” was observed. 3. Juice of batata diluted with a moderate quantity of water and of perfectly neutral reaction was boiled for a minute, whereby all power of reactions disappeared. After 24 hours, the power of giving the reactions was regenerated. Prof. Loew observed that the oxidase reaction could thus be three times ‘‘ regenerated.” 4. A moderately diluted radish juice was boiled for a few minutes : After 24 hours; sow oe no regeneration. sk ASs, Bsworcieise . eee peroxidase, red and violet reactions appeared slightly. c 4 GaSe: ks a12s ee tee the same. 5. The same tests were carried on with milk which was boiled for a few minutes : Srehy i meee no regeneration. sa~ NAST S5a0 oo eine ERE oe all original reactions reappeared, but weaker, a AiGAaYS, Awitiea ci cin ete eee the same. 6. The water extract of cured tobacco leaves boiled for a few minutes was tested as follows: After 24 hours: no regeneration. ea Comet trace of peroxidase and red reaction reappeared, the latter stronger than the former. 1 Also this author jinferred the existence of corresponding zymogens, These facts as well as the observations of S/ow!zoff (Z. physiol. Chem, 31.) and Efstein (Arch, Hyg, 30.) seem to fully establish the enzym nature of the oxidases. Recently, however, Kas/Ze and Loewenhart (Amer, Chem, Journ, 24) have tried to prove them to be organic peroxides and supported their view by showing that these also are very sensitive towards such poisons as kill enz | On Oxidizing Enzyms in the Vegetable Body. 233 After 3 days: Peroxidase and red reactions reappear- ed weak. » §5days: The two reactions mentioned appeared ; also a faint trace of oxidase and the violet reaction. 7. For the ‘‘regeneration” of catalase, the following experiment was made: 100c.c. of beer yeast paste was heated to 85°C. to kill the catalase. After 3 days standing in presence of some ether the result upon addition of hydrogen peroxid was as follows : Oxygen developed. After 5 min. After 30 min. PeTEG Eerie 4.0% win ohare dive. cain’ 2Os1 \C.C. 30.9 C.c. RACE Nie an cs ow pom aft Ee, AS ie.c: ‘“‘Regeneration”’ of Catalase seems to have taken place ina small degree but further experiments are necessary to draw a safe conclusion. In general, the existence of zymogens of oxidizing enzyms appears to be highly probable. The duration of the heating will of course much influence the result, since zymogens also will gradually thus be destroyed. Separation of the Oxydizing Enzyms from each other. Since the nature of the oxidizing enzyms is not quite fully established, isolation meets with some difficulty. Prof. Zoew has already shown that oxidase and peroxidase are not nucleoproteids, but behave like albumoses. All oxidizing enzyms may be precipitated with alcohol, hence the fractional precipitation with alcohol might be of some value, as_ the following experiments will show : One volume of the juice of radish root was mixed with two volumes of absolute alcohol, whereby a white precipitate was produced. With the filtrate all reactions were obtained with the exception of that of oxidase, The precipitate, however, dissolved in a little water, yielded an intense oxidase reaction, but the other four reactions above-mentioned also were obtained. 234 kK. Aso: Upon addition of three volumes of absolute alcohol to the radish juice, the filtrate failed to show the reactions, while the precipitate yielded all the reactions very strong. Therefore, a separation of the peroxidase from oxidase is possible only in the first case mentioned. A similar observation was made by Behrens with the juice of tobacco-leaves. Upon mixing with double the volume of strong alcohol a filtrate was obtained, which gave only the peroxidase reaction. All oxidizing enzyms tested are precipitated by saturation with ammonium sulphate. The juice of radish was saturated with ammonium sulphate. The filtrate did not yield any color reactions while the residue contained the substances which produce all color reactions caused by the oxidizing enzyms. On addition of 19§ acetic acid to the radish juice, a small quantity of a white precipitate was obtained, but all reactions could be obtained with the filtrate. Hence the substances which cause such color reactions belong in all probability to kinds of albumoses and not to the nucleoproteids. Another method of separating peroxidase from oxidase may be based on the behavior towards sodium fluorid and sodium silicofluorid, but much caution is here necessary, since after the early destruction of oxidase, the peroxidase is also gradually attacked by sodium fluorid and sodium silicofluorid.! General Conclusions. 1. Various vegetable objects which yield the well known guaiac reactions for oxidase and peroxidase, also yield a red reaction with guaiacol and hydrogen peroxid. 2. Storch's reaction on milk with paraphenylendiamine and hydrogen peroxid is also obtained with many vegetable objects. Generally a green color appears first, but in certain cases it changes soon to violet, while in most cases, this change is very slow. 1 Tused 5% solution of sodium fluorid and a saturated solution of sodium silicofluorid and tested after well shaking the mixed solution, On Oxidizing Enzyms in the Vegetable Body. 235 A new reaction for an oxidizing enzym was found, which consists in the production of a deep violet color on the addition of tetrame- thylparaphenylendiamine and hydrogen peroxid. This reaction is obtained with various vegetable objects. This new reaction may be used to distinguish fresh milk from Loiled one. The spermase reaction found by Gréiss is also obtained with several seeds, resting as well as germinated ones. In the case of resting seeds, only the embryo yields this reaction. The red guaiacol reaction is caused by a separate enzym which is more stable then even the peroxidase.! Sodium fluorid and sodium silicofluorid interfere with all the color- reactions. Oxidase is killed sooner than the other oxidizing enzyms. The green and violet reactions might be caused by enzyms different from oxidase and peroxidase, sinee their killing temperatures lies between that of oxidase and peroxidase. This influence becomes also probable by the comparison of the resistance power to injurious compounds. Sugars do not interfere with the color reactions caused by oxidizing enzyms in any notable degree. Neither interfere soluble egg albumin and pepton. However tannin interferes very seriously. The presence _of zymogens of the oxidizing enzyms is very probable. The separation of peroxidase from oxidase may be accomplished by adding two volumes of absolute alcohol to one volume of a plant juice. Hereby oxidase is precipitated while the main part of the other oxidizing enzyms is present in the filtrate. The oxidizing enzyms which produce these color reactions resemble albumoses. ey oo J fp ‘ - | ; es i : 2 i = i 3 : fis ay 4 fe : = * 5 ' ; ; E ed i) t a . ; > “ . ’ - ‘ On the Curing of the Kaki Fruit. BY S. Sawamura. The fruit of Dospyros Kaki L. is an article of food extensively consumed in Japan. The flesh of this fruit contains glycose and fructose, while the reserve carbohydrate in the seeds is mannan.! The fruit was Sweet variety. | Astringent variety. EP IPR re Gas 20 shite Swe Gwe a%e.0-4. 5 82.03 83.65 Mitrogenous matters .........:.. 0.61 0.58 COS 9 SUE ie ie oe Renee 0.02 0.02 Sugar and other N free extract .. 13.62 12.56 ecmanducernels 2 of... sec va) 3.29 2.76 ee Mere In Tere, a.0 o6 oS ae ods 0.43 0.43 There exist sweet and astringent varieties of this fruit. In the unripe state both contain much tannin, but only with the former variety this tannin, and consequently the unpleasant taste, disappears in the ripening process. _ This change is brought on by oxidizing enzyms which act on the “tannin as Aso* has shown. As to the astringent variety artificial means ~ are resorted to, to remove the unpleasant taste. These means are: I. Keeping the fruits for 12 hours in a barrel containing vapors of 1 Jshit, These Bulletins, Vol. Il. No. 2. 1896. 2 According to the analyses of the Tokyo Sanitary Exp, Station. 8 Gerber found that when this fruit is allowed to ripen in a confined atmosphere, it yields 10% of ethyl alcohol, mixed with other alcohols, among which amyl alcohol was copious, and further that 2 the aromatic principle was a mixture of amyl and ethyl acetates with traces of oenanthylates and pelargonates. (Greer : Fermentation p. 351.) + Botanical Magazine, Tokyo, 1900, 238 S. Sawamura: On the Curing of the Kaki Fruit. alcohol. Generally Sake-barrels just emptied are used for this purpose. II. Keeping the fruits for 12 hours in warm water. III. Subjecting the pealed fruits to a dessicating process in the sun. It will be seen at once that the methods intend to kill the cells. Hence there can be no doubt that the disappearance of the tannin taste can not be due to the action of the protoplasm. I have kept fruits for control in a flask containing vapors of chloroform, and also in this case the tannin taste, which had disguised the sweetness of the fruits, disappeared. My quanti- tative tests further showed that the change did not consist in the trans- formation of tannin into sugar.2. There remains, therefore, only one conclusion in regard to the effect of the curing process, and this is that the tannin is changed to a tasteless substance by partial oxydation brought on by the oxidizing enzyms present in the fruits. These emzyms are confined to the cytoplasm, while the tannin to the vacuole. By killing the cells the osmotic properties of the cytoplasm are changed, and the oxidases can now pass into the vacuole, where they can mix with the cell sap and exert their action upon the tannin. 1 30°-40° C, are sufficient to effect the change. 2 For this purpose a ripe fruit, which had still an astringent taste, was divided into two equal parts, one of which was cured by exposing to vapors of chloroform, while the other left without any treatment, In the former 55.919 of sugar but no tannin was found, while in the latter 56.189% of sugar and 8.239% of tannin were contained. —~ —_ On the Different Forms of Lime in Plants. BY K. Aso. In the quantitative determination of mineral constituents of plants no attention was thus far paid to the different forms in which these compounds occur in the plants. The usual way to incinerate the plants before the analysis of the mineral constituents was carried on, did not permit any distinction. But, since the occurence of different forms may be of consider- able interest, I made some determinations of lime and magnesia in this direction. Lime may occur in plants 1) as salts easily soluble in water, 2) as salts difficult soluble in water, but easily soluble in dilute acetic acid and further 3) as salts insoluble in water and dilute acetic acid, but soluble in hydrochloric acid. In this last mentioned case, only calcium oxalate comes into consideration. Finally 4) compounds of lime with organized matter may occur, from which the lime also can be extracted by dilute acetic acid but not by boiling water. Since the juices of plants have generally a more or less acid reaction and since I applied a relatively very large amount of water in the first extraction, the second form of lime salts will probably be almost entirely removed by the first treatment with a relatively very large quantity of boiling water. The plants serving for my investigations were collected in the morning while poor in starch and kept in darkness for a few days until the iodine test showed that the last portion of the starch was consumed. This was done to reach comparable results, since the amount of starch varies so greatly in different periods of the day that the results of the analysis would be too much influenced by it. As objects were sclected: 240 K. Aso: 1. Potato. (Leaves and stems, collected before flowering, on Nov. 9. 1900). ; 2. Buckwheat. (Leaves and stems, collected after ripening, on Nov. 8. 100). 3. Wild clover. (Leaves and stems, collected before flowering, on Apre26. 190m) 4. Barley. (Leaves and stems, collected before flowering, on Apr. 26. 190). 20 grams of air-dried finely powdered substance were extracted twice with one liter of boiling water,! and the residue thoroughly washed with hot water until every trace of soluble lime was removed. This residue was extracted with one liter of 5% acetic acid in the cold for 24 hours, frequently shaking the mixture, filtering and washing the residue with distilled water until the filtrate had lost every trace of acid reaction. The residue thus obtained, was treated with one liter of 5% hydrochloric acid for 24 hours with frequent stirring.? In each of these extracts the lime and magnesia content was determined separately. The results obtained were as follows: In tco parts of dry matter, Objects. CaO, soluble in: Total. Water Acetic acid aha ga 1 Fo} til] £0) donee heoepapedsto. 0.332 0.875 1.586 2.793 Buckwheat pone-cewce: 0.056 0.367 1.524 1.947 * Glovers 3 ee eee 0.858 0.742 0.489 2.089 Barley oe eee 0.438 0.259 trace 0.697 1 ‘The aqueous solution had a slight acid reaction in each case, 2 In the final residue, lime and magnesia were absent or present only in very minute traces, CaO MgQ which shows that magnesia plays also here a very important réle in the fruiting process. % The lime factor for buckwheat before flowering is 3, while in the time of fruiting it is 1.3, On the Different Forms of Lime in Piants. 241 Iu 100 parts of dry matter. Objects. MgO, soluble in: 7 Total Water Acetic acid | Hydrochloric acid, | | 1.617 0.550 0.217 2.384 1.050 1 0.417 trace | 1.467 | 0.491 0.162 trace : 0.653 0.307 0.094. | trace | 0.401 We see from this table, that the quantities of the different forms of lime vary considerably. In the case of potato and buckwheat, only a sma!l quantity of lime compounds soluble in water are present; more lime is found in the acetic extract and still more in the hydrochloric acid extract. Much calcium oxalate is produced in these leaves, while those of barley? contain only a trace of it. As to the magnesium compounds, they are either soluble in water or in acetic acid. Only a trace of magnesia remained in the residue. Church's investigation? with albino leaves have demonstrated that lime is more abundant in the green leaves than in the white. In order to see whether this pathogenic albinism shows a chemical analogy to the normal albinism, I have determined the lime also in the white and grcen ‘ parts of the leaves of Avundo Donax separately.* 1 Probably present as secondary phosphate, in which form it is not poisonous for the nuclei. T have made also analogous determinations of lime in maize-stalks with the following result : In 100 parts of dry matter ; soluble in Water, Acetic acid. Hydrochloric acid. CaO 0.130 0.128 trace MgO 0.241 0.120 trace But, as these stalks had been dried and were exposed to the rain on the field after harvesting, the result is not a normal one. 2 Calcium oxalate is absent in most Graninee, 3 Journ, Chem, Soc, 1878 and 1886. 4 This separation by scissors was made as carefully as possible, nevertheless it was not absolutely complete. 242 K. Aso: On the Different Forms of Lime in Plants. The analysis was carried out in the way above mentioned. In 100 parts of dry matter, total ash : White -parts:: 77 —se% Seem nce nek eee) «oe eee Green parts res ware oe tees ee ee, Se eee CaO soluble in | | + areca Water Acetic acid Hydrochloric acid. White parts ............ 0.213 | 0.216 trace 0.429 (reeni pants meaeaeeete 0.313 0.226 trace 0.539 MgO soluble in Total, Water Acetic acid Hosa acid, Witte pairtsieeasemeneee 0.387 0.068 — 0.455 Greens paises 0.444 0.069 — 0.513 White parts. Green parts. Lime-factor > Gini ve. pees 0.9 Tei From this table, it is clearly seen that the total ash and lime-content of the green parts exceed those of the white parts, and also that the lime- CaO MgO smaller than 1. Hence the inference that the amount of lime increases factor in the green parts is larger than 1, while that in the white with that of the chlorophyll bodies, other things being equal, has again found confirmation. 2 > it oe oS On the Alcohol Production in Phenogams, LY T. Takahashi. Since the interesting discovery of E. Buchner that the expressed juice of yeast can cause the alcoholic fermentation of glycose and fructose, the question has been raised, whether zymase is also produced in phznogams, .and whether the alcohol production in the process of intramolecular respiration of cells of phanogams is due to zymase or to the action of the protoplasm itself. Former observations made by Srefe/d! were not in favor of the assumption of zymase. He had observed that the expressed Jutce of grapes the surface of which had previously been sterilized, did not show any alcoholic fermentation while the sterilised ¢xtact grapes them- selves formed some alcohol by intramolecular respiration, like many fruits rich in sugar do. Recently Godlewskt published an exhaustive investigation on the intramolecular respiration of the pea.? He stated the interesting fact that peas and other seeds kept under water can form considerable quantities of alcohol.? In order to decide whether the living protoplasm itself or zymase causes this alcohol formation, God/ewshi ground the peas to a fine powder, whereby the protoplasm would be killed, but zymase can remain intact. In this condition hardly a trace of carbonic acid (and consequently also no alcohol) was formed during two days,* while the same weight of 1 Landw. Jahrb. 5. (1876). 2 Bulletin de l’Academie des Sciences de Cracowie, 1901. 8 The pea seeds seem to produce more alcohol than many other seeds. Goeverwsty mentions: ,, Die Menge des Alcohols, welche bei der intramolecularen Athmung der in reinem Wasser liegenden Erbsen- samen sich bildet, kann bis zu 22% der urspriinglichen Trockensubstanz der Samen erreichen.” 4 Later on bacterial action was noticed, 244 Y. Takahashi: entire seeds produced 5 cc. carbon dioxid in 24 hours. But although this result apparently proves the absence of zymase, Godlewskz hesitated to draw this inference, and declares among other things: ,, Es ware méglich, dass durch das Zerreiben der Pflanzenmasse irgend welche Substanzen aus gewissen Zellen frei gemacht werden, welche, sei es durch Nieder- schlagen der Zymase, sei es auf andere Weise, die Wirkung derselben auf- heben.” I have made the following experiments in regard to the intramolecular respiration of the pea. One hundred seeds, weighing 33,3795 g. in the air dry state were left for one hour in a 1 per mille solution of mercuric chlorid in order to destroy, all adhering germs, then washed with sterilized water and trans- ferred to a sterilized Erlenmeyer flask nearly filled with sterilized water and connected with a small flask containing baryta water. The room in which this flask was now observed for 38 days showed mostly on average a temperature of 16°, sometimes however only 9°. Four days after the start of the experiment little bubbles were observed rising from the peas and this slow development was continuous with the exception of those days, on which the temperature had sunk to 9° During the 38 days of observation, the water above the peas had remained perfectly clear, proving that the original sterilization was perfect. A careful microscopical examination of the surface of the peas at the close of the experiment also proved the absence of mucor, yeast and bacteria. A determination of alcohol by distillation yielded 2 grams, calculated from the spec. grav. of the destillate at 17,5°=0.99646. The iodoform test proved further that the alcohol was indeed ethyl alcohol. This result fully confirms God/lewskt’s observation. The quantities of alcohol, however, were larger in Godlewski's experiment, what can be accouuted for by fhe higher temperature (17,4-24,7°) in the latter case. 3 A further test proved that a number of the peas from the flask had stzll retained their germinating power. The water, further, in which the peas had remained and from which the alcohol had been distilled off served for the determination of the solid matter it had extracted from the peas. The residue weighed 1.146 g.=4,01% of the dry matter of the a On the Alcohol Production in Phanogams. 24 wr peas.4 Of this nearly one fourth consisted of mineral matter. In order to decide whether zymase was the cause of the alcohol? production, the skin of the peas was removed and kernel and skins separately placed in a sterilized 10% solution of glycose and kept at 31°C. Even the smallest quantities of zymase would thus have been betrayed by some development of carbon dioxid, but xot a single bubble was noticed even after one day. 1 must infer, therefore, that symase Zs absent and cannot be the cause of the alcohol production in the intramole- cular respiration of the pea.* Protoplasm.itself is the producer, but it works very much slower than does zymase, and at a temperature of 31°C. seems to stop that action. A few words may here be permitted in regard to the relation between the intramolecular and the normal respiration. Goedlewski arrives at the view: ,,Die intramoleculare Athmung im Sinne der alcoholischen Giarung bildet unter normalen Bedingungen aller Wahrscheinlichkeit nach das ~ erste Stadium der normalen Athmung in allen denjenigen Fallen, wo sicl dieselbe auf Kosten der hydrolysirbaren Kohlenhydrate volizieht.” This view seems to me not justified. In the first place we would have to assume two different ways of respiration one for fat, another for sugar; the fat would be capable to be burnt up directly, the sugar not directly but only after transformation into alcohol. This would contradict all our conceptions in regard to the chemical character of fat and sugar. The latter is much easier oxidized than the fat, why should the protoplasm not be capable to oxidise sugar directly, but the fat? To transform the sugar into alcohol before the combustion takes place would not only be entirely superfluous but would render the respiration more difficult ; it is, e.g., well know that sugar is oxidized much quicker in the animal organism than alcohol is. In the second place it would follow from God@/lewshi’s view, that those seeds that can produce more alcohol by intramolecular 1 The fresh peas contained 28.55 %% of dry matter, 2 It may be mentioned here that Afasé has found some alcohol in pea seedlings that had germi- nated under normal conditions at 24°C, for 48 hours, The view of Z7roné that zymase exists in the pea (quoted by Green, Fermentations) finds no support by my experiments. 246 T. Takahashi: On the Alcohol Production in Phenogams. respiration should also show a more energetic normal respiration, the pea should excell therefore many other seeds, what is not the case as far as our knowledge gees. There may be other products formed in other seeds from sugar, when they are forced to intramolecular respiration, as fat, lactic acid, etc.— There exists certainly a connection between the normal and the intramolecular respiration but this relation is different from that entertained generally. Doubtless a high degree of chemical energy exists in the living protoplasm. When this is transferred upon the imbedded molecules of sugar, a certain lability is produced in them which leads to direct combustion when free oxygen is present, but to various other decomposi- tions when free oxygen is absent. This is in a few words the theory of O. Loew. 1 Cf. O. Loew» Die chemische Energie der lebenden Zellen (Chapter XII), Mtinchen 1899. = —- 69> ee Can Alcohols of the Methane Series be Utilized as Nutrients by the Green Plants ? BY S. Sawa. While alcohols in moderate concentration act poisonously on the higher plants, as Zswkamoto has shown,! it seemed to me’ probable that in proper dilution various alcohols might bea nutrient for them. It is known that methyl and ethyl alcohol occur in small quantities in certain green plants. Methyl alcohol was observed by Gufze?t as well as by Magquenne in the distillate obtained from juices of Evonymus, Hedera, Lolium, Urtica, Galium, Helianthus, Syringa, Dahlia, Acorus, Heracleum, and Pastinaca. Ethyl alcohol» has been found in Heracleum, Pastinaca and Axnthrts- cus. P. Mazé has observed that ethyl alcohol is a normal product of vegetation in the germination of seeds. He found alcohol, for instance, in germinated peas kept for 48 hours at 24°C.2 He thinks that this alcohol was formed from glucose by a kind of fermentation process in the cells. It has been known long ago further that alcohol may be formed in the interior of sweet fruits. It has also been shown by Sofkorny that plant-cells can form starch from methylic alcohol under the influence of light. This fact renders it probable that the starting point for the preparation of starch in the leaves is the formic aldehyde, the next 1 Journal of the College of Science Tokyo, 1895. 2 Compare also Ged/ew shi, Jahresber, f, Thierchemie 1897, p. 700. * In darkness this process does not take place, 248 S. Sawa: oxidation-product of methyl atcohol.1 It seemed to me of interest to compare, therefore, the action of methyl alcohol on the growth of the plant with that of some other higher alcohols. For the following experi- ments served young onion plants grown from the seed which were kept at first in 0.1% solutions of methyl, ethyl, butyl, and isobutyl alcohols in order to observe, whether there were any poisonous actions in that dilution, and since after 10 days no injurious action was observed, they were now placed in the following nutrient solution which was obtained by mixing 10% solutions of the nutrients in the following proportions : 45 cc, Calcium nitrate, 15 ,, Magnesium sulphate, 24 ,, Potassium nitrate, 6 ,, Mono-potassium phosphate, trace of ferrous sulphate. 5 cc. of this mixture were added to 100 cc. of the 0.1% solutions of the alcohols on the 26th of March. The length of the leaves was measured at the same time and the solutions renewed as often as a_ turbidity due to the development of yeast and bacteria was observed. There was soon a considerable difference, as seen from the following table contain- ing the results. The experiment lasted for 29 days. Ihe letter 9o “in the table signifies the leaves present at the starting of the experiment ce while the letter ‘‘n” signifies the new leaves formed. The temperature of the room varied between 12-22°C. The flasks containing the plants were placed ona table well exposed to the diffused day-light, but not to the direct sun-light.2 Some of the leaves dried off gradually at the tips and’these dried-off parts were not considered in the measurement. 1 Kinoshita has further found that methylic alcohol can be used by green plants for the formation protein (Bul. College of Agriculture, Tokyo 1895 vol. II. No. 4.) Zeezw had observed before, that it can be used by bacteria as food, even in absence of other organic material, 2 Perhaps the nutrient effect of assimilation under very bright light would have obscured the nutritive effects of methyl alcohol. ee ee ne Can Alcohols of the Methane Series be Utilized as Nutrients by the Green Plants? > 49 Length on March 16th. Length on March Increase in absence of 26th, mineral nutrients, Alcohol, ee Individual Individual |Summed | Absolute popes sae leaves cm, Summed up, leaves cm, 1 | cm, Relative % et eee en Ree —_—_—_—_——————— —_—_—_—_—_————_. (o) 21.5 | | ° 18.3 55-9 A eo 3H | J | JO Methyl! alcohol = ba) LL | oe eee ———— (0) = aifes B : | 14.6 37-7 o’ 19,2 OW 27.7) (in) A 4.6 | 10.4 of ers Ethyl alcohol meted —— Oo 24.8 Beye sage, 48 98 4 Ot: TG oO 16.5 | A ‘ 5.0 16.7 O23 55 , Butyl alcohol pee | ee | Se ee | fo) 18.2 B ; 7.0 24.6 oe 10.2 7 j | 9 20.1 | hes m3 | 377 e 9.9 (3) Tsobuty! alcohol 2 eee o 8618.2 | , 35.4 7.1 20,1 “: eo 6172 ? oe x80 | ; a o dead 0 14.9¢4)]) A 28.4 o” 268 10.4 36 of 13.5 n 12.0 Control Pee a Oo 24.5 | : 31.7 40.5 $8 27.7 B fo ine x o” . 16.0 : sk Ethyl alcohol Butyl alcohol Control I | 250 Alcohols, A Methyl alcohol B A WW We Length on April 24th, Individual leaves cm, MS ] — dead 30.2 29.0 25.0 EO, 20.5 11.5 (5) 535 clead 26.0 | dead dead 60.5 Summed up S. Sawa: 79.7 Absolute cm, Increase after mineral nntrients were given, Aleeeys OZ Relative % 135.6 Remarks, (5) A bud was formed on April 17th. (1) 1.7 cm on the tip had dried off. (2) 3.0 cm on the tip had dried off, (6) A bud was formed on April 18th. (3) 5.0 cm on. the tip had withered. (4) 3.3 cm on the tip chad died off. (7) A bud was formed on April 2oth, (8) A bud was formed on April 19th. F’ " _ Se) ete ee ; 7 Can Alcohols of the Methane Series be Utilized as Nutrients by the Green Plants? 257 The result is therefore that methyl alcohol in 0.19¢ solution has acted as a nutrient and this was even plainly visible during the first period of the 10 days when no mineral nutrients were added so that the mineral nutrients previously absorbed had come alone into play. Ethyl alcohol had less nutritive value while butyl] and isobutyl alcohols had a retarding effect on the growth.1 1 The higher alcohols are also more poisonous tor plants than the lower as 7’sz/amoto had already observed, —_——~ er = . = f - . os it = : 7 _ » - — 1 2 = ne oo ee 2 he 7 ; | > = ; : ne On the Occurrence of Mannan. C. Kimoto. The peculiar horn-like consistency of the seeds of Lrachycar pus excelsa, a palm tree, frequently Serving in Japan as an ornamental tree, led me to the supposition that it might be rich in mannan, especially since the test with iodine showed the absence of starch. Although small quantities of mannan have been observed in many seeds, there exist on the other hand not Many cases in which it forms the only or the principal reserve carbohydrate in seeds. Instances of this kind are the nuts of Phytelephas and the seeds of Diospyros Kahi.! Recently Bourquelot and H. fférissey observed a considerable amount of-mannan in the seeds of Phoentx Canariensis. Of considerable interest is further the observation of L'sijz that the root of ¢ onophallus Conyaku (Amorphophallus Rivier 2), used in Japan as an article of food, is exceeding- ly rich in mannan. Tsukamoto has shown further that this plant contains also mannan in the leaves and stalk.’ Of special interest is the fact discovered by Gabriel Bertrand‘ that while xylan is present in the wood of Angiosperms it is replaced in the Gymnosperms by mannocellulose. The wood of Abies pectinata yielded 9.6% mannose, on being boiled five hours with hydrochloric acid of 5°. Gnetacee which from the transition between the Gymnosperms and the Angiosperms gave thus no or very little mannose. In order to test the seeds of Zyach yearpus excelsa for mannan, the seeds were finely cut up after removing the shells, and 2.07 ¢.(=*1. 49 g. 1 Ishii, Bul, vol. IL No, 2 of Imp, College of Agriculture, University of Tokyo, ® Ibid page 103 and Atnoshita, Ibid No, 4. * Ibid, vol, IIT, * C.r,, vol. 129, page 1023. 254 C. Kimoto: On the Occurrence of Mannan. dry matter) boiled for three hours with sulphuric acid of 39%, replacing the water lost by evaporation. The filtrate was neutralized with barium carbonate, the liquid evaporated to moderate concentration and acetate of phenylhydrazine added, which produced a voluminous crystalline pre- cipitate. After standing one day it was collected on a filter and recrystal- lised ; it melted at 190°C. and weighed 0.822 g., corresponding to 0.528 g. mannose and or 0.475 g. mannan, and to 31,3699 of the dry seed. The observation of Aertrand mentioned above on the occurrence of mannan in coniferous trees led me to look for mannan also in the wood of Cryptomeria. 1 boited small chips of the wood (dry matter=9Ig.) with dilute sulphuric acid of 3% for three hours, and proceeded as above, whereby I obtained indeed a hydrazon which had all the properties of mannosephenylhydrazon. From the quantity obtained—10,2 g.—it follows that the wood of Cryptomeria contains 6,359 mannan. The seeds of Rhodea Faponica were also examined for mannan, since starch is absent in them. After removing the shells and pulverising the seeds, the treatment was essentially the same as above mentioned. The analytical data are as follows: Original substance so45-% shed ate ee ee 11.2444 2. Dry amattet «4 s2)4 aac ape eae a 8.7141 g. Mannose phenylhydrazon .=.....). ope 21252 ee. corresponding to 1.3645 g. mannose. Hence this result shows that the dry matter of the seed of Rhodea Faponica contains 14.28 % mannan. While I was occupied with the investigation of the reserve carbohy- drates in the seeds of ducuba Faponica, I noticed in a recent number of Comptes Rendus de l’Académie des Sciences, November 25, 1go1., that Mr. Champenots had made an investigation on the same object and found in these seeds galactan, mannan and pentosan. A similar investigation was published in the first December number by G. Duédat, who found mannan also in the seeds of some Li//aceae. ee a nate ee On the Genera! Occurrence of Bacillus Methylicus in the Soil, BY T. Katayama. A very important function of bacteria in the soil is the oxidation of organic matter, whereby carbonic acid is produced which may partly be absorbed by the root and carried to the leaves for assimilation, and partly serve to dissolve carbonates and phosphates of lime and magnesia, and thus facilitate the absorption of certain mineral nutrients by the roots. Many kinds of bacteria have thus far been observed to occur in soils, but one kind of bacterium which is present in the air of Japan as well as in Europe has not yet been looked for in soils, it is Bactllus methylicus which is obligate aerobic.! This microbe has the characteristic faculty to assimilate salts of formic acid and certain related compounds, as oxymethyl-sulfonate and methyl-sulphate of sodium and methyl alcohol, all containing only one atom of carbon in the molecule. The isolation, therefore, of this bacillus is rather simple, since the other microbes thus far known cannot subsist on formates. Bacillus methylicus occurs frequently in putrefying liquids, as can easily be ascertained by infecting from such liquids, a culture solution containing $%% of sodium formate as the only organic nutrient. Bacillus methylicus® alone will thus develop, forming reddish films. Since it appeared of some interest, to ascertain whether this exquisite aérobic t This microbe was first observed by O, Zoew, Central Blatt. f£. Bakteriologie. 1892. 2 According to the nomenclature of A.B. Zefmann this name would have to be changed to Bacterium methylicum, since it forms no spores, 256 T. Katayama: microbe is of general occurrence in soils I have examined soils from different parts of Japan in this direction. The samples of soils were well shaken with water (50 gr. with 100 c.c.) and 10 c.c. of this liquid was added to the following solution, that had previously been heated to 100° C, Sodium: formater prsewics ese tee ae 0.5% Dipotassium phosplate-—=..40 ees ener OLA Diammonium phosphate 4.4.6 450 eeee Gis Magnesium: sulphate. 27 er. 0a O:8L 5, In some cases a thin growing film of red color made gradually its appearance, especially along the rim of the solutions, while in other cases the film was very pale or white. Of this a second infection was then made into other flasks containing also this solution, in order to exclude the other soil bacteria, that had been suspended in the to c.c. soil extract applied in the first solution. A plate culture on gelatin was finally prepared and from a colony thus obtained the following tests for identification were made. 1). On potato; slight red elevated colonies, giving a wormlike appearance above the streak. 2). In bouillon it grows in the form of a coherent skin which on shaking sinks to the bottom; the bouillon liquid itself remains clear. 3). Instab culture, it grows only on the upper surface of the canal formed, not in the depth, liquefying the gelatin on the surface gradually. 4). On a gelatin plate, the colonies grow in elevated round forms and of a very slight reddish color, and begin to liquefy the gelatin after a few days. 5). In the formate solution, the cell has the form of a short straight rod, generally 1 yz. thick and 2-3 y. long, but in bouillon and on gelatin, it grows longer and shows sometimes the form of the comma bacillus. | In a number of cases, I have observed, however, in making the gelatin plate culture from the second infection of the formate solution that besides red colonies more or less white colonies of the same character also appeared. i in a6 ae 4 @ 2a oie On the General Occurrence of Bacillus methylicus in the Soil. 257 On inoculating from these white colonies in bouillon and making potato-cultures, I observed that this white bacillus resembled in every respect, except the color, to the red Pacillus methylicus, and I think it highly probable, therefore, that this is a variety of it. I examined altogether 20 soils from the depth of 3 cm. and found the Bacillus methylicus in every instance. The results are seen from the following table ; Locality. This College, Komaba. bby ” ” Noda, Shimosa province, Near the river Tone, Musashi province. Kumagai, Musashi province, Kumagai, Musashi province, Kind of soil. light loam \ iB (mulberry plantation) | 2 light loam (experimental field, without manure for 14 years) light loam | I (forest) | ot fertile loam (1 (farm) (2 sandy soil (1 (rice field) lo fertile, clayey loam {1 (farm) | 2 un-manured, clayey \t loam 4 (near rail road) lo Date. (were inoculated in the formate solution.) 14 January 18 January * This sample was taken from the depth of r em. t This sample was taken from the depth of 5 cm. Color of film. (after 2 months.) red slightly reddish (few samples) white (many samples) white slightly reddish white white 258 TT. Katayama: On the General Occurrence of Bacillus methylicus in the Soil. Locality. Kind of soil. fertile loam. I Ueda, Shinano province. (mulberry plantation) ( 2 un-manured gravelly i I ” ” ” soil (mulberry plantation) | 2 Nagano, fertile, clayey loam. { I eae sce (mulberry plantation) | 2 Date. (were inoculated in the formate solution) 19 January ss fertile, clayey loam, (1 Nagano, Shinano province 7 Shinano pro : (forest) 5 z clayey loam, I age < ee ea % (experimental field, a iS aa a without manuring {[ 2 for many years) Kawasaki, fertile, sandy loam, | I Musashi province. } (orchard) = fertile, sandy loam. Kyoto, d y Yamashiro province, (forest of oak trees) Clor of film, (after 2 months) white ” ” ” 1i March red It was noticed very clearly that manured fertile soils yielded also the Bacillus methylicus ina greater number than sterile poor soils, to judge from the great difference in the time of film developement in the formate solution. Since this bacillus occurs in putrefving manure, in the air and in every one of the examined soils, it can be concluded that it is of general OCCULENGE: I intend to make further investigations in regard to this microbe and to its colorless variety. | _ On the Liquefaction of Mannan by Microbes. BY S. Sawamura. Since I had repeatedly occasion to observe the loss of viscosity of a certain mucilage used in the manufacture of Japanese paper, I was led to examine the action of microbes on mannan. The mucilage in question was derived from Hydrangea paniculata Sieb. var. minor, and consisted to a considerable extent of mannan containing also some araban and galactan. The paper :manufacturer, being not acquainted with bacterial action could not explain the rapid loss of viscosity when the diluted mucilage was kept for some time.! My examination of such spoiled mucilage soon revealed the presence of numerous micro-organisms to which no doubt also was due the observed production of acidity. Since the mucilage in question contained chiefly mannan and since we know that galactan is not liquefied by bacteria as the experience with agar cultures has demonstrated long ago, it seemed to me of considerable importance to test various kinds of microbes upon the power of liquefying mannan jelly. In Japan occurs in commerce a food called ‘' Konnyaku,” prepared from the root of Conophallus Konyaku by treating with dilute milk of lime, and resembling starch paste, which consists almost exclusively of mannan.? For my experiments I prepared a much more diluted product dissolving 3% of the refined dry preduct of the said root ina hot solution containing 199 of pepton and 0.19% of magnesium sulphate 1 This calamity was easily avoided by antiseptic means, which the writer proposed. ® Tsuji. Mannan as an Article of Human Food. Bulletin of this College. Vol. II. No. 2. p. 103. Atnoshita, On the Occurrence of Two Kinds of Mannan in the Root of Conophallus Konyaku, bid, Vol. II. No. 4. p. 205. Tsukamoto, ibid. Vol, 11, No, 7. p- 406. 260 S. Sawamura: and dipotassiumphosphate. This solution gave on cooling a transparent jelly resernbling the agar jelly, used for bacteriological cultures. After the mixture was sterilised in the usual way it was infected with various kinds of bacteria and yeasts, and kept at 36°C. for two days. The results are seen from the following table. Names of the Microbes infected. Liquefaction of Mannan Jelly. Saccharomyces cerevisiz. -- 2 apiculatus, pas from the mucilage. = 39 Micrococus from Koji. ~ Streptecoccus from silk-worm. - Bacillus capsulatus, = ” cyanogenus, = Tfueppe. Fs ” » megatherium, = s, | Mmesentericus ruber. —- vulgatus. + 39 ” pyocyaneus. 5 prodigiosus., , typhi murium Zenkerei » subtilis. ” On the Liquefaction of Mannan by Microbes. 261 It was only Baecllus mesentericus vulgatus of the sixteen species tested that liquefied mannan in two days, the same microbe which also can saccharify starch. But according to my observation it does not saccharify araban. The above named microbe cultures were observed for several weeks further, but it was only Bactllus prodigiosus that showed a weak action on mannan within that time. The careful examination of the spoiled mucilage mentioned above convinced the writer, that the loss of viscosity was caused by Bacillus mesentericus vulgatis. But this phenomenon is considerably accelerated when a certain wild yeast propagates luxuriantly in the mucilage.! This influence is difficult to explain, since this yeast in itself has no action on mannan and further does not ferment the mannose formed by the bacterial action.? l infected sterilised mannan jelly with Bacillus mesentericus vulgatus alone, and in a second case with this bacillus and that wild yeast together, and ina third case with that bacillus and beer yeast. After keeping the jelly at 36°C. for some days the sugar formed was determined with Fehling's solution. -The results were as follows :— Sf 4 eee in 07 WiaroWes: Length of Strength of Sugar found in 9% Sanex, Culture, Solution, of mannan. - | | | Vulgatus, 4 days. | 10% 13.782.% = Vulgatus + the Wild Yeast. = | 17.675 .% 28.23% Vulgatus, 2 days. | 5% 5-742.% == 1 | Vulgatus + the Wild Yeast. f. | 8.614.% | §5002% = ——— 7 ae —— ‘| == = Vulgatus, I day, 5% 3.676% — Vulgatus + Beer Yeast, as | fe 4.923% 38.93% 1 The accelerating action of yeast on diastase was observed by Morris, Central-Blatt fiir Agricul turchemie 1902. p. 286. 2 0,04 vol. % of alcohol was formed by cultivating it in 109% glucose solution at 36°C. for 12 days. S. Sawamura: On the Liquefaction of Mannan by Microbes. i) O* bo The amount of alcohol formed from mannan by beer yeast alone was so minute that it could not be quantitatively estimated. Further investigations are necessary to explain satisfactorily the accelerating action of the yeasts on the liquefying action of Pacitllus mesentericus vulgalus. Summary. While thus far no microbe was observed liquifying galactan, there exists an exception as regards mannan, since Saczllus mesentericus vulgatus can easily liquefy mannan jelly. Laecllus prodigtosus appears to contains also traces of this enzym. I must here express my thanks to Prof. Dr. O. Loew for his useful suggestions made to me during this and other investigations, and to Prof. Dr. Y. Koza¢ who kindly provided me with the pure cultures of bacteria used in this experiment, and also to Mr. 7. Yamasaki, Assistant of this College. ee ee a Chemical Note on a Singular Phenogamic Parasite. BY T. Suda. In the province of Tosa and in the southern part of Kiushiu in Japan a phznogamic parasite frequently develops on the roots of Symplocos and allied plants. Sometimes it occurs also in the Idsu province and around Nikko. Of this interesting phenogam, however, only female plants have thus far been found in Japan. In Tosa and Kiushiu people prepare from it a sticky mass called ‘ Torimochi’ resembling bird lime, by steeping the rhizom in water and crushing it well. This mass is of black color. This singular plant is a kind of Balanophora but not identical with Balanophora dioica occuring in India and Java where people dry the plant and use it directly like candles for illuminating purposes, since it contains a resinous substance in considerable quantities. F. D. Hoocker* states that Balanophora dioica in India has not the long branching rhizomes and not so much resin-as the related Balanophora elongata growing in Java. Th. Poleck published some studies on this latter species and found the melting point of the resin (Balanophorin) go-95° C. Several points in regard to the Japanese species of Balanophora seemed to me of some interest, it was the amount of resin present and further, the amount of lime and magnesia, since phenogamic parasites require less lime than green plants. The material furnished to me contained 20.95% dry matter and this yielded 7.819§ of ash. The amount of the resinous compound present, extracted by ether, was 15.87% of the dry matter. For determination 1 Transactions of the Linnean Society. Vol, XX. Part I. (1863). 264 T. Suda: Chemical Note on a Singular Phenogamic Parasite. of lime and magnesia in the upper part 10 grams of dry matter were incinerated and the analysis carried out as usual. I obtained: CAO Pere oe one esi dans oe 0.129% MEO recess cyt a Se eee 0.244% This shows that also this phaenogamic parasite like the well known Cuscuta is poor in lime compared with green plants and that the amount of magnesia is larger than that of lime, while in the leaves of green plants the reverse is observed. ————— “a. we On the Action of Formaldehyd on Pepsin. BY S. Sawamura. Formaldehyd exerts an injurious action on enzyms, as was first observed by O. Loew.4 Pepsin and diastase are killed in one day when left in a neutral 599 solution of formaldehyd. Pottevin? observed an injurious action on rennet and sucrase, the latter being injured in already one hour at 54°C. by a formaldehyd solution of even less than 59. At low temperature, however, sucrase resists formaldehyd more than other enzyms do, as Sokorny? observed. This author* also stated that a formaldehyd solution of 19% kills maltase in 24 hours, one of 59% in half an hour, and that one of 0.5% prevents the action of rennet. Catalase is killed in one hour by a solution of about 4% of formaldehyd. Bliss and Novy® stated on the other hand that pepsin and diastase are not injured even after weeks by diluted solutions of formaldehyd, and this statement as far as it relates to pepsin was corroborated by Peke/- haring™ who writes: ,, Mit Formal zu einem Gehalt von 2-3% versetzte Loesungen von Pepsin in Salzséure kénnen ohne merklichen Verlust an verdauender Wirkung Tage lang aufbewahrt werden.” ,, Beim Priifen muss der Gehalt an CH,O mittelst Verdiinnung oder Dialyse herabegesetzt werden, damit nicht das Fibrin selbst fiir die Verdauung ungeeignet gemacht werde.” 2 Journ, f, prakt. Chem. 37, p. 104, (1888). 2 Annales de I’ Institut Pasteur 8, p. 796 (1894). 8 Pfliig. Arch, 85, p. 267. 4 Ibid. 5 Loew; Report No, 68. U.S, Dept. of Agriculture (1901), 6 Journ, of Exper, Med. 4, p. 47 (1898). 7 Z, physiol, Chem. 35 p. 29 (1902). 266 S. Sawamura: On the Action of Formaldehyd on Pepsin. These statements, contradicting apparently the observations of others, induced me to make some experiments with pepsin. Fife grams of the commercial pepsin were dissolved in 50 c.c. of a 10% solution of formal- dehyd, while another 5 g. were dissolved in distilled water containing a little thymol. After 24 hours standing both solutions were precipitated with strong alcohol and the precipitates after well washing with alcohol dissolved in water containing 0.2% hydrochloric acid. These solutions were kept with some fibrin? at 36°C. for 24 hours with the result : Normal pepsin : Fibrin dissolved completely. Pepsin treated with formaldehyd . Fibrin not attacked at all. In a second experiment the mucous membrane of a hog’s stomach was digested with four times its weight of 0.2% HCl for a week. To one part of the filtrate was now added 20% formalin (8% formaldehyd) while to another the same amount of water with some thymol. After one day these solutions were tested as above with the same result. The result of my first experiment is more decisive than that of the second, since the adhering formaldehyd had been carefully separated from the pepsin before the fibrin served for the experiment. Two causes may be responsible for the discrepancy between my results and those of Pekelharing. In the first place, my formaldehyd solution was of higher concentration, and in the second place, that author tested a solution of pepsin in 02% hydro- chloric acid, while I had a perfectly neutral solution. It may be that the hydrochloric acid protects just those labile groups which otherwise enter in combination with formaldehyd. —- <0> -—- 1 This fibrin after being freshly prepared was kept in glycerol, Before application it was kept for some time in very dilute hydrochloric acid and washed, oai2P 108. Wie eas <., ., Ueber die Einwirkung des Hara-Brennens, VON O. Shishido, Ringakushi. Einleitung In alter Zeit war das japanische Inselland vermuthlich vollkommen mit Wald bedeckt, und die Bewohner suchten ihren Lebensunterhalt im Walde; Friichte miissen ihnen ganz unentbehrlich gewesen sein. Aber mit der Zunahme der Bevélkerung kam zuerst die Frage der Ernahrung derselben ; man musste also seine Lebensmittel kiinstlich gewinnen, und so entstand unsere Landwirthschaft; daraus folgte dass die Wald-Bestinde, welche sich in den Ebenen und Gebirgen befanden, mit der Zeit nach und nach abnehmen mussten; ja die Bestiande wurden als ein Hinderniss fiir die Landwirthschaft sogar vielfach angesehen. Da diese Abholzung grosser Arbeit und Zeit bedarfte, so fing man an, die Bestiinde durch das Feuer zu vernichten; Verbrennung des Waldes und Fillung der Bestiinde dauerten bis in spatere Zeitalter fort. Auf diese Weise entstanden die heutigen kahlen Gebirge und bestands- losen Ebenen, welche wir “ Hara’ nennen. Diese Fliche an Hara ist in Japan ausserordentlich gross, die folgende Zusamenstellung gibt uns eine ungefahres Bild davon: (December 33 Meiji, 1900) Staatswaldungen ......... Rape eks Att oixkee. 13072602 cho MERE MMACLUTICCOTD 5 , cs cuits wah a el ck iG ache de «. 2091784 chd Privatwaldungen, peremeindewaldiingen, }: .24.cc.<\asscescdvovcennses 743C091 chéd deer Waldungen SRST AIO hds Siy yas ehh ectide 646d 1305 Jak ah scene toy ERE ERT I CAO 268 0. Shishido: Staats-Hara 2... cess Wi eid sa cng eta ACCC ee Krone arias: 2 Sed ee eon sec ace ee a A ee Privat- Hara ‘Gemeinde = avi 2a... eae ee tS ach 1053462 chd ee andere Hara SUNT Lee ae tere ener eee eee er eee .. 2045302 cho — (i cho=oo172 Bs) demnach betragt die Flachen-Summe der Hara in Japan 2645302 ché. In Japan herrscht die Sitte, jahrlich das Hara-Gras zu brennen; es gehért somit zu den wichtigsten Fragen, die Erfolge des Brennens zu untersuchen, besonders interessirt dieses Thema den Forstmann, der vielfach unter dieser Brennsitte zu leiden hat. Aus diesem Grunde untersuchte ich die Vegetation, die Boden- und die Wuchs-Verhaltnisse auf verschiedenen Hara. Fir meine Arbeit fand ich hinreichenden Stoffin der Gegend der Kiyosumi-Schulwaldungen. I. Abschnitt. Die Geschichte der “ Kiyosum-Hara.” yy) Wie erwahnt, haben ausgedehnte Wald-Bestande in alten Zeiten das ganze Land bedeckt; diese Walder wurden aber vielfach abgeholzt oder abgebrannt. Solche Zerstérungen des Waldes dauerten bis in die neuesten Zeiten herein; Ebene und Gebirge sind solcherart in grossem Massstabe in Kahlflichen (Hara) umgewandelt worden. Die Hara, welche nun auf diese Weise entstanden ist, benutzt man hauptsichlich zu landwirthschaft- lichen Zwecken, namlich zur Gewinnung von Grasdung, oder gebraucht sie als Wiesen; der letzte Fall kommt aber in Japan sehr selten vor. Die genante Kiyosumi-//ara ist getheilt in zwei Geimeinde-Hara, d. h. jene von Amatsu und von Tojo und einen Theil, weleher Ga den Schuiwaldungen gehért. Von diesen Gemeinde-Hara’s existiren keine urkundlichen Nachweise. Niemand kennt deren Entstehungs-Geschichte ; nach der Ansicht der Gemeindevorsteher in Amatsu und TOjo sind diese Hara nicht in derselben Weise entstanden wie die meisten Hara in Japan Ueber die Einwirkung des Hava-Brennens, 269 sich bildeten. In der Zeit, als die Schogun aus dem Hause Tokugawa iiber das ganze Land herrschten, bestand in den meisten Gegenden die Sitte, kahle Gebirge dem Privatbesitz zu tiberweisen ; aber in den Provinzen Awa und Kazusa (wozu die Kiyosumi-Hara gehort) existirte diese Sitte nicht. Diese Gemeinde-Hara sind in den altesten Zeiten noch teilweise mit Bestanden bedeckt zu denken und verblieben der Gemeinde als Freigiiter. Man benutzte die Hara hauptsichlich, um eine Grasart, sogenanntes “Kaya” (Miscanthus sinensis Anders) zu gewinnen; um ein mdglichst gutes Wachsthum desselben zu erzielen, brannte man diese Hara, jahrlich. Nur tiber die Geschichte der Hara, welche zu den Schulwaldungen gehort, hat man einige Kenntniss; der Wald an Sannodai und derselbe an der Nordseite von Suzuriischi, also die jetzige Hara, sind im Jahre 2 Tempo ginzlich abgeholzt worden, seitdem sind sie szezmal gebrannt worden, einmal im 24 Jahre Meiji (1891) und das zweitenmal in Meiji 32 (1899). Die Gemeindevorsteher in Amatsu und Tojo sagten mir, dass man sich jetzt wiederum entschlossen hat zur Aufforstung dieser Hara. Ii. Abschnitt. Zustand der Kiyosumi-Hara. I. KAPITEL: SAodensustdnac. A) Lage und Fliche. Die Kiyosumi-Hara umfasst drei Hara, nimlich jene im Schulwald, dann jene von Amatsu, Uchiura, und endlich die Hara von TOjo, Seijé, Hiroba und Hamaogi; die ganze Flache der zwei letzteren Gemeinde-Harea hat 391,0325 a; die Flache der in den Schulwaldungen gelegenen Hara betriigt ca 20 ha. Diese Hara befindet sich an der Nordostseite von Awa, angrenzend an die Provinz Kazusa; das Bergland, auf welchem diese Hara liegen, erstreckt sich von dem Platze der Schulwaldungen gegen das Meer zu. Der Lage nach gehdért diese Hara wie Prof. Honda bestimmte, der subtropischen 270 QO. Shishido : Waldzone an. Diese Hara ist auf Gebirgsausliufern steiler Ausformung, welche bis zum Meere reichen, ausgebreitet und hat keine nennenswerthe ebene Fliche aufzuweisen. B) Boden. (Grundgestein) Der Boden, aus welchen die Halbinsel Awa gebildet ist, besteht hauptsachlich aus drei Gesteinsarten: Tuff, Schieferthon und Sandstein. Diese Gesteine sind im Allgemeinen rauh und weich, leicht verwitterbar und bilden viele Bodenmodificationen. Th KAPITEL:) Das tke: Das Klima ubt natiirlich auf den Pflanzenwuchs einen grossen Einfluss aus. 1. Die Temperatur. Der ‘‘Kuroshio”” oder warme Meeres-strom, welcher an die Siidost- Kiiste der japanischen Insel, von Siidwest nach Nordost bespillt, tibt auf die Lufttemperatur einen grossen Einfluss aus; namentlich wirkt er auf das Klima der Halbinsel Awa in héchstem Grade ein, weil sie sich in dem Ocean hineinstreckt ; es ist also kiihl im Sommer, warm im Winter, d. h. es ist das Klima ein sogenanntes Seeklima mit abgestumpften Extremen. 2. euchtigkert Die Halbinsel Awa hat eine grosse Luft-Feuchtigkeit, wodurch die Regenmenge ausserordentlich anwachst, im August erreicht die relative Luftfeuchtigkeit ihr Maximum und im Februar ihr Minimum. a: wDer.Vebel. Wo warme und kiihle Str6mungen des Meeres sich mischen, wie an der Ostseite der Halbinsel, werden hiaufig dichte Nebel erzeugt; diese treten meist in den Monaten April bis September auf. Ueber die Einwirkung des Marc-Brenucus. 271 Bit BOSE. Er kommt in Kiyosumi sehr selten vor, Frosttage sind in Kiyosumi in einem Jahre nur 20 und zwar meist in der Zeit von Ende Oktober bis Anfang Mirz. 5. Der Wind. Wind hat auf den Pflanzenwuchs eiren grossen Einfluss., deshalb muss sein Einfluss beachtet werden. a) Hauptwind. Von April bis September (5 Monate) herrscht Westwind, wihrend in den 7 anderen Monaten sich hauptsichlich Nordost oder Nordwestwind einstellen ; im Allgemeinen sind Nordwinde die Hauptluftstrémungen. 6) Sturm. Es gibt in dieser Gegend oftmals Stiirme; nach den Untersuchungen in Choshi und Fura (32, Meiji, 1899) herrschen die “Aeftigex Winde” meist in Winter, wahrend in Februar, Marz, Spitwinter und Anfangs des Frihjahres “ Stirme” auftreten. Das Klima ist in Kiyosumi fiir Menschen und Pflanzen giinstig zu nennen, da die Temperaturdifferenzen zwischen Sommer und Winter sehr gering und die Feuchtigkeitmengen, welche zum Pflanzenwiichse nétig sind, reichlich vorhanden sind. WII. Abschnitt. Cultur der Gegend bet Kiyosumi. Auf der Halbinsel Awa und Kazusa befinden sich zahlreiche niedrige Hohenziige, zwischen denen in den Thiilern eine recht miissige Fliiche an Ackerboden vorhanden ist. In der Kiyosumi Gegend, wo das Hiigelland seine grésste Héhe erreicht, findet man Ackerland nur an der Kiiste in geringer Ausdehnung ; es ist eben kein Land vorhanden, welches vortheilhaft landwirthschaftlich bo “J iS) 0. Shishido: benutzbar wire, wenn auch hie und da die Landwirthschaft bis zum Berghange sich ausdehnt, wo dessen Gefalle minder steil ist. Die Flachen der Hara des forst-und landwirthschaftlich benutzten Bodens_ beziffern wie folgt sich: Namen. Ackerflache. Waldflache. Zarafliche, Summa. pik Kamogawa 198.31 ha. 10.84 ha, 8.36 ha. 217. 5ite las 38 % Amatsu 149533) 45 981.20 ,, ZT SslOianss 1448.72 ,, ZEON oes Kominato LOS:35 "G5 sia S. se 192-27) 2, TO52:470 us, 18:35, Tojo 395.88, 1180.45 5, 22507 Tees 1805.04 ,, 1207) es Seij6 300.40 ,, 50740) Us O23 5h. QIO.15 5, TW 2 G5 Kameyama 544.19 ,, HiR2-5OlGs 390-6250, 2088.31 ;, ES: 7 35 Kururi PGR | ef 1527.64 ,, 20.4010 TS83121:, 1-69 15; Omi FAQS? 4; BA OT sa: 69.48 ,, ACTRE aie 1ytaae Oikawa 262.46 ,, WASOLGOR 211 el BT 7-4 5a le 233 0:27a las EOTGS Ges Nischihata 65:92 3: 904.99 ,, 1992.01 _ ,; 2554192, 3 stom Otaki 333-83» 33:70? 1; 2520O 659:63)'h: 326)1 3) Fusano STiye aaa MO332551 | 35 476.84 ,, 2328510) 15; 20:5) 53 Fusamoto 447.81 ,, DOTA ier, 19240) o3, o3niGn ie 2ONTE ss Katsuura 204.53: HOU-S3) 159.90 ;, S557 tess 287i 0s Seikai 2AOMGMEEs 58617 Gur. 140.03; 22:97; EGS. 53 Ueno 569.34 5» 709350) 953 49.00 ,, IQS7203) ss Owes, Man sieht, dass der Waldgrund im Allgemeinen die Ackerkulturflache an Grésse iiberwiegt, namentlich hat Amatsu eine Waldfliche von 67,7%, daher ist auch die Hauptbeschiftigung der Leute dortselbst die Verkohlung des Holzes, Holzschlag, Holztransport u. s. w. ferner bemerkt man die grosse Ausdehnung der H/ara; diese Hara werden zwar theils zur Futter- und Kayagewinnung benutzt, aber thatsichlich geben die “ mzezsten” Theile der //ara in diesen Gegenden nahezu keinen Ertrag. to Ueber die Einwirkung des Hara-Brennens. NI oS) IV. Abschnit”. Die Hara und thre Besttzverhiltnisse. I. KAPITEL: Besttzarten - (A) Der Universitét gehérende Hara. Ein Theil derselben befindet sich in Kiyosumi und anderer liegt in Kiwada, an der Nordseite der Schulwaldungen; die ganze Fliiche der Univ. Hara beziffert ca 20 ha; sie soll jedoch baldigst in Wald umgewandelt werden. Unter diesen Hara gibt es einen Theil, die Hara Musadogadai, deren Benutzungsrecht den Leuten von Kiyosumi vergiins- tigungsweise zur Gewinnung der Kaya (Miscanthus sinensis Anders uberlassen wird, dafiir miissen sie dann der Universitit gewisse Gegenleistungen stellen, (B) Die Gemeinde-Hara. Den Haupt-Theil der sogenannten Kiyosumi-Hara nimmt die Gewreinde- Hlara ein, von der zwei Arten unterschieden werden: erstens die Hara von Amatsu und Uchiura deren totale Fliiche 318 ha. betrigt, sweitens jene von Tdjo, Seijd, Hiroba und Hamaogi mit einer Fliiche von 88.9 ha. Die erstere bildet die sogenannte Mukodmine-Hara welche nérdlich an die Schulwaldungen grenzt, und sich nach den Kiisten Amatsu und Uchiura erstreckt; die zweite Hara reicht von den Schulwaldungen gegen Tojo hin. Il. KAPITEL: Gewinnung der Haraprodukte. Auf der Gemetde-Hara kénnen die Berechtigten nach ihren Bediirfnissen, den Anwuchs der Hara ernten, es gibt keinerlet Beschrinkung darin, sei es dass Jemand seinen Ernteantheil an Gras verkaufen oder einem Andern unentgeltlich iiberlassen will; nur die Gewinnungszeit der Produkte ist etwas beschriinkt; es muss niimlich die Gewinnung der Kaya im December, und jene der anderen Griiser, von Juli bis August statt- finden; in der der Universitiit gehérenden Hara ist die Gewinnung dieser 274. 0. Shishido: Griiser principiell wzcht gestattet, nur in Musadogadai, welches ebenfalls zu den Schulwaldungen gehért, kénnen die Leute in Kiyosumi, nach Erforderniss Kaya bekommen, gegen gewisse Gegenleistungen. V. Abschnitt. Das Brennen der Hara. TI. KAPITEL: Zweck des Brennens. (A.) Entstchung aus alter Sitte. Die Entstehung der Hara ist im Allgemeinen dem Niederbrennen des Waldes zuzuschreiben, man wollte diese Flaiche eben nicht wieder Wald werden lassen und brannte sie deshalb von Jahr zu Jahr, um Gras als Futter und Diingungsmittel zu bekommen, namentlich war Kaya das Haupt- produkt. Die Sitte des Harabrennens dauert bis Heutzutage, so werden z. B. die meisten Theile der Kiyosumi-Hara noch jetzt jahrlich gebrannt. Der Vorsteher in Amatsu sagte mir, dass die Leute in diesen Gegenden der Ansicht huldigen, dass die Pflanzen der Hava durch das Brennen kraftiger werden. (B.) Landwirthschafthche Benutzung der Hara. Da wie erwiihnt in diessen Gegenden nur gering Menge an Ackerflache sich vorfindet, so bedarf es hier wenigen Grases als Diingmittel; auch erhalten die Leute grosse Mengen Fischdiinger, welcher fiir den Acker- boden wirksamer ist. (C.) Werth des Harabrennens sur Gewinnung vou Futtergriasern. In Kiyosumi u. Umgegend wird eine grosse Zahl Rinder gehalten welche fiir den Transport des Holzes nicht entbehrt werden kénnen ; man bedarf fiir den Unterhalt dieses Viehes viel Futtergras, welches allein auf der Hara gewonnen wird. Um nun Griser fir Futterzwecke in méglichst grosser Menge zu erhalten, brennen die Bauern die Hara alljahrlich. ee Ueber die Einwirkung des 77ava-Brennens. 275 II. KAPITEL: Verfahren des Harabrennens. Wer Haragras brennen will, hat dies bei dem Gemeindevorsteher anzumelden, welcher dann Nachricht davon an die Polizeibehérde gibt. Wenn die Erlaubniss ertheilt ist, beginnt man mit dem Brennen des FHlaragrasses, nachdem die Absicht u. den Zeitpunkt des Brennens den benachbarten Bodeneigenthiimern mitgetheilt wurde. Ill. KAPITEL: Zeit des Harabrennens. Die Ernte des Kaya (Miscanthus sinensis) erfolgt in jener Jahreszeit, wo das Kayagras seine maximale Hohe erreicht hat, das ist in der Regel Mitte December der Fall; die iibrig bleibenden Griiser auf der Hara brennt man dann im Monate Februar; bald nach dem Brennen begriint sich die Hara wieder, und schon Ende April ist die Hara mit neuem Graskleide bedeckt. IV. KAPITEL: Die Methode des Harabrennens. Sobald die Polizeibehérde dem Eigenthiimer einer Hara die Erlaubniss zum ffarabrennen gegeben hat, beginnt derselbe mit dem Brennen der Hara an einem stillen Tage, wobei ein Polizei- und ein Gemeinde-Beamter sowie der benachbarte Bodeneigenthiimer gegenwartig sein sollen. Auf den Grenzlinien stellen sich Leuten an um benachbarte Hara oder Wald vor dem iiberlaufenden Feuer schiitzen zu kénnen. VI. Abschnitt. Phystkalische Eigenschaften des Bodens. I. KAPITEL: Jn Kiyosumi. A) Ttefe. Die Tiefe des verwitterten Bodens ist nach Ortsverhiiltnissen verschieden, nimlich in den Thiilern ist er tiefer als auf den Bergen, weil der Boden am Abhinge nach und nach thalwiirts herabgefiihrt wird durch Regen und andere atmosphiirische Niederschlige. Die Tiefe der Verwitterungsschicht welche ich auf verschiedenen Probefliichen untersucht habe ist folgende : 276 0. Shishido: Namen der Probefliche. | Fliiche. | Neigang Standort. darchsehnittliche ‘Tiefe. Kiridéschi (Nordseite) | 20 [). m. 250 am Mittelhang 0.29 Mm. (Stidseite) - eee al " Ola Gy as Kajisaka (Nordseite) : B9n) al ae 0.41 ,, 2 (Sudseite) 53 ga + 0285; Omiyama _(Nordseite) . 34° C275 I. (Stidseite) ss | 34° 2 0.50 4; bs 2. (Stidseite) > 36° : OATTA Sannodai (Nordseite) . 35° +: OTe; bs (Siidseite) A. Saiet || zs OOIy. Suzuriischi (Nordseite) = 32° z) O23: = (Stidseite) = 356 | s 0.29 >» Musadogadai (Nordseite) 93 B20 =: ©3533; 2 (Stidseite) * OD el * 0.44 > Nanamagari (Nordseite) 3 250 . 0.45 3 & (Nordseite) A Boone | * @.02 Sengen =| = (Stidseite) be 34° ONTiaeres durchschnittlich. mitteltiefgriindig 0.434 m. Immer wird man auf oft gebrannter Hara einen seichten Boden finden, wihrend der selten gebrannte Haraboden oder Waldboden mzztteltzef- bis tiefgriindig ist. Auf Grund meiner Untersuchungen komme ich zu dem Schlusse dass der Haraboden in Kiyosumi im Allgemeinen mitteltief-griindig ist. B) Bindigkett. Die Bindigkeit des Bodens ist in den einzelnem Theilen der Hara etwas verschieden, je nachdem gewisse Parthien der Hara jihrhich oder pertodisch gebrannt wurden; die zéhrlich gebrannten Theile haben lockeren oder grob gekritmelten Boden, wihrend auf selten gebrannter Hara die Struktur feiner und gekriimelter ist. a eee eee i) NS] NS Ueber die Einwirkung des Hara-Brennens. Im Allgemeinen aber kann man den Boden auf der Kiyosumi-Hara wegen der Luftfeuchtigkeit als ziemlich mild oder locker ansprechen. C) euchtigkett. Die Hara, deren Bodeniiberzug jahrlich gebrannt wird, miisste cigentlich starker ausgetrocknet sein, weil der Boden dfter und langer direkt der Luft ausgesetzt ist; aber er ist dennoch durchweg ziemlich feucht, der Grund davon ist ebenfalls darin zu suchen, dass die Luftstr6mungen auf dieser Halbinsel an sich sehr feucht sind, so dass disser Unterschied nicht so fithlbar wird. II. KAPITEL: Awssere Zustinde der Kiyosumi-Hara. Die Hara in Kiyosumi wurde frithe schon von Wald-Bestiainden entblosst, jetzt ist die ganze Flache derselben hauptsichlich von Kaya Grass (Miscanthus sinensis Anders) bedeckt. Aber die Menge der Kaya ist natiirlich je nach dem Wiederholungszeitraume des Brennens verschieden ; in allen jenen Theilen, welche zeniger oft gebrannt werden, mengen sich Baumarten (wie hauptsichlich Quercus glandulifera, Quercus serrata, Quercus acuta, Quercus vibrayeana, Machilis japonica, Evonymus europeus var. hamiltonianus, Rhododendron indicum var. Kaempferi, Salix Sieboldiana, Spiraea callosa n. s. u.) und Halbbiume den Miscanthusarten dev. Aus diesem Umstande kénnen wir leicht erkennen dass die Hara, wenn man sie fir langere Jahre in Ruhe liesse, wieder zum ihrem eigentlichen Urzustande kommen, d. h. in Wald sich umwandeln wiirde. So kann man z. B. in Nanamagari, wo die Hara seit 10 Jahren wicht gebrannt wurde, foleende Baum- und Halbbaumarten beobachten : Quercus glandulifera Quercus serrata Evonymus alatus var. subtriflora Spiraea callosa Eurya Japonica Diervilla glanduliflora Viburnum dilatatum 278 Q. Shishido: Machilus Thunbergii Clethra barbinervis Smilax china Quercus acuta Quercus myrsinaefolia Rosa multifiora Rhododendron indicum var. Kaempferi Rubus palmatus Litsea hypoleuca Salix Sieboldiana ete: Auf“ alvdéhrlich” gebranntener Hava konnen nur Grasarten vorkommen, welche ich in einem spateren Abschnitte besonders anfiihren werde. Ill. KAPITEL: Das Verhdltniss swischen Boden und Klima. (Verwitterungs process) Der Boden ist das Verwitterungsprodukt der Grundgesteine, und die Verwitterungsgrésse hangt von dem Klima ab, welches auf dem betrachteten Platze herrscht. Die Verwitterungsschicht bedeckt den Boden in den Gebirgen bei Kiyosumi in sehr verschiedener Dicke. Da der Boden der Kiyosumi //avra aus den Gesteinen der Tertiarperiode gebildet ist, so sollte die Verwitterungsschicht eigentlich ziemlich bedeutend sein, aber die Wirklichkeit zeigt uns eine grosse Ungleichkeit des Harabodens. Er ist in seiner Verwitterungsschichte verschiedentlich sehr flachgriindig, namentlich ist dies am Kamme und in oberen Hange zu bemerken; der Grund hiezu liegt wohl in der Steilheit der Gebirge, wodurch bei Regengiissen die verwitterten Bodentheilchen, welche keine geniigend schiitzende Pflanzendecke haben, abgeschwemmt werden. Diese Erschein- ung muss natiirlich auf der Hara intensiver als im Walde sein, weil der Haraboden direkt frei der Atmosphire ausgesetzt ist, wahrend im Walde der ganze Boden durch die Kronen der Baume eine Uberschirmung erfahrt. Die Verwitterungsschichtendicke wurde in einzelnen Haratheilen folgendermassen gefunden : Uober die Einwirkung des ara-Breunens. 279 Namen, Jahre des Harabrennens, eect Mittel. Kiriddshi (Nord seite) alljahrlich gebrannt | 69.6 m. m. 3 (Stid seite) = 3 | Taek sat 33 | Ea) Kajisaka (Nord seite) a sf Pe 52S ox wy te | 60.0 ,, 3 (Stid seite) fe rs (5 fe ae cha Omiyama (Nord seite) fiir drei Jahre nicht gebrannt By} 2, | ] “ yan i | 121.2 Nanamagari (Nord seite) fiir 10 Jahre nicht gebrannt | FIO] 33 | 110.7 Musadogadai (Nord seite) tiber 10 Jahre nicht gebrannt 98.1 | 08.1 Sengen (Nord seite) Urwald | SOAl aa ss | ‘ AS (Stid seite) i 97.2 : sade (D’e Neigung der einzelnen Hara ist in der vorgegangenen Tabelle bereits bezeichnet worden.) Die Verwitterungsschichten u. damit der produktive Boden werden auf der ungeschiitzten Hara nach und nach immer seichter durch die Abschwemmung der feinen Theile thalwirts. Die Niitzlichkeit u. Not- wendigkeit der Verwitterungsschicht fiir den Pflanzenwuchs brauche ich kaum weiter zu besprechen. VII. Abschnitt. Vergleich der Effekte des Brennens der Hara in verschiedcnem Brenn-Terminus. Um den Einfluss des brennens Hara besser zu beobachten ist néthig, dass man ausfthrliche Untersuchungen auf verschiedenen Heara’s macht. Die meisten Theile der Gemeinde-Hara in Kiyosumi sind, wie ich schon 220 Q. Shishido: erwihnte, aljahrlich gebrannt, wahrend die der Schulwaldung gehérende ¢ oO Hara nur sehr selten ebrannt wurde; die Hava in Kiwada, welches auch der Universitat gehért, wird ebenso alljahrlich gebrannt. Ich untersuchte nun folgende Theile : i 9 6. lod fe Kiriddshi und Kajisaka. Omiyama. Omiyama (Siidseite) und Musadogadai (Siidseite). Suzuriischi (Siidseite). Sannodai und Suzuriischi (Nordseite). Nanamagari. Musadogadai (Nordseite). alljahrlich gebrannt. seit 3 Jahre nicht eebrannt. seit 6 Jahre nicht gebrannt. iiber 8 Jahre nicht gebrannt. bis 24 Meiji (1891) alljahrlich und spiater im Jahre 33 Meiji (1900) gebrannt. seit 10 Jahren nicht gebrannt. iiber 10 Jahre nicht gebrannt. ausserdem habe ich den Urwald Senge zum Vergleich herangezogen. Um I. KAPITEL: Priifung muttels Probeflichen. den Antheil der verschiedenen Grasarten an jeder Hara festzustellen, habe ich Probeflichen von 2 L|. m. genommen, und gebe nun die Mittel aus je drei Proben. Zahl der Kiridodshi (Nordseite). (alljahrlich gebrannt) (Neigung = 35°) (Datum 10/4 u. 25/4) lateinischer Name. Miscanthus sinensis Anders, Sanguisorba officinalis L., Senecio krameri Fr, et Sav. Thalictrum minus L, var, elatum Lecoy. Arundinella anomala Steud. Plectlanthus glaucocalyx Max, var, japonicus Max. Hikiokoshi japanischer Name. Grdser % Susuki . 318 38.6 Waremok6 80 9.7 Yaburegasa | 74. 9.0 Akikaramatsu 67 8.1 Todashiba 51 6.2 37 4.7 os wie Ueber die Einwirkung des Hava-Bronnens. Lespedeza bicolor Turcz. Saussurea Tanakae Fr, et Sar, var, phyllolepis Max. Pteris aquilina L, Potentilla fragarioides L, var. ternata Max. Lysimachia clethroides Duby, Viola silvestris Kit, var. gryposeras A, Gr, Serratula atriplicifolia B, et H. Dioscorea Tokoro Makino, Polygonum cuspidatum S., et Z,. Cirsium japonicum DC. Atractylodes lyrata S. et Z, Carex breviculmis R, Br. Poa trivialis L, Seseli libanostis Koch, var. daucifolia DC, Cirsium spicatum (Max). Euphorbia Esula L, Manettia ignita (Vell), Heteropappus hispidus Less, var, isochactus Fr, et Sav. Viola japonica Langsd. Allium japonicum Rgl. Aster indicus L, Artemisia vulgaris L. var. indica Max. Disporum sessile Don, [Hagi] Tohiren Warabi Mitsubatsuchiguri Okatoranoo Tachitsubosumire Kumatoribokuchi Onidokoro Itadori Noazami Okera Aosuge Himeichigotsunagi Ibukibéfu Yamaazami Hagikuso Kwaens6 Yamajinogiku Ko sumire Yama rakkyd Yomena Yomogi H6chakusod Kiriddshi (Siidseite). (alljahrlich gebrannt) (Neigung = 33°) (Datum 10/4 u. lateinischer Name, a Miscanthus sinensis Anders. Imperata arundinacea Cyr, var, Koenigii (Benth). 25/4) japanischer Name, Susuki Chigaya Griaser in 2{]. m,| 344 _— 173 ~~) Zahl der | : o- £2 Y. Shishido: Brachypodium silvaticum R. et S. Yama kamojigusa 140 12.1 Inula salicina L, Kasens6 64 5.5 Miscanthus sacchariflorus Hack. Ogi 62 5.4 Sanguisorba officinalis 1, Waremok6 54 4.7 Lysimachia clethroides Duby. Okatoranoo 40 a5 Viola Patrinii DC, var. chinensis Ging, Sumire 36 3.1 Serratula atriplitifolia B. et H. Kumatoribokuchi 35 3.0 Pteris aquilina L, Warabi 33 2.8 Smilax china L. [Sankirai] 25 7 | Thalictrum minus L, var, elatum Lecoy, Akikaramatsu 24 2.0 Athyrium nipponicum Bak. TInuwarabi 18 1.6 Senecio Krameri Fr, et Sav. Yaburegasa 16 1.4 Plectranthus inconspicuus Mig. Yamahakka 16 1.4 Euphorbia Sieboldiana Mor, et Dec Natsutodai 15 1.3 Lespedeza Sieboldi Miq. [Hagi] iS) Cirsium spicatum Max. Yama azami 7 Aster indicus L. Yomena 6 } Polygara japonica Houtt. [Hime Hagi] 6 Patrinia scabiosaefolia Link. Ominaeschi 6 Dioscorea Tokoro Makino, Onidokoro 5 3.6 Potentilla fragarioides L, Kijimuschiro 5 Polygonum cuspidatum §, et Z, Ttadori 5 Vitis Thunbergii S, et Z. Ebizuru 4 Seseli Libanostis Koch, var, daucifolia DC, Tbukibofu 4 Gymnadenia conopea R, Br, Chidoriso I Aus den Tabelien, ersicht man “dass auf jihrlich gebrannter Hara” hauptsachlich die folgende Grasarten auftreten : Miscanthus sinensis Anders. 1157 100 Sanguisorba officinales L. Serratula atriplicifolia B. et H. Senecio Krameri Fr, et Sav. Pteris aquilina L. to CO Ww Ueber die Einwirkuag des Hara-Brenuens, Thalictrum minus L. var. elatum Leroy. Polygonum cuspidatum S. et Z. Imperata arundinacea var. Koenigii (Benth.) Potentilla fragarioides var. ternata Maxim. Brachypodium silvaticum R. et S. etc. Diese Grasarten sind vorzugsweise Lichtgrdser, d. h. sie konnen meist nur auf nacktem Boden gedeihen, oder kommen wenigstens 6fter u. vorzugs- weise in nahrungsarmem Boden vor. Auf der Hara, welche seit dre? Jahren nicht gebrannt ist, sind dagegen die folgende Grasarten anzutreffen gewesen. Omiyama (Nordseite). (fiir 3 Jahre nicht gebrannt). (Neigung = 34°). (Datum 11/4 u. 26/4). 27 ) i ; ; Zahl der | lateinischer Name. japanischer Name. | Gr, in y 4 2} m. Miscanthus sinensis Anders, Susuki 295 | 25.2% Smilacina japonica A. Gr, Yukizasa | Sr | 69 Aster japonicus Miq. Yamashirogiku 65 | . 56 Carex breviculmis R, Pr. Aosuge + i oe Artemisia vulgaris L. var. indica Max. Yomogi AE itn Be Potentilla fragarioides L, var, ternata Max, Mitsubatsuchiguri | 39 3:3 Cypripedilum japonicum Thunb, Kumagaiso 37 3-2 Plectranthus inconspicuus Miq. Yamahakka 34 | 2.9 Cirsium japonicum DC. Noazami | Sas. To ae Petasites panies Miq. Fuki 31 | 2.6 Carex conica Boott. Himekansuge | 25 2.2 Viola Patrinii DC. var, chinensis Ging, Sumire 20 | 1.7 Kraunhia floribunda (Willd) Taub, Fuji 18 1.5 Viola silvestris Kit. var, grypoceras A, Gr, Tachitsubosumire 17 | 1.4 Polygonum cuspidatum S, et Z. Itadori 17 1.4 » 284 0. Shishido ; Lysimachia clethroides Duby. Okatoranoo 16 1.4 Euphorbia sieboldiana Mor. et Dec. Natsutodai 16 1.4 Thalictrum minus L, var. elatum Lecoy. Akikaramatsu 16 1.4 Crepis japonica Benth, Onitabirako 16 1.4 Dioscorea Tokoro Makino, Onidokoro 15 1.4 Poa trivialis L. Hime ichigotsunagi 15 fea Lactuca squarroba Miq, forma indivesa Max, Honba akinonogeschi 15 leg Polygara japonica Houtt. [Himehagi] 15 Te Erigeron annuus Pers, Himejoon 15 eel Aspidium dissectum Met. Hoshida 15 13 Disporum sessile Don, Hochakus6 14 1.2 Disporum pullum Salist. Tochikuran 14 1.2 Cirsium spicatum Maxim, Yamaazami 14 ior Clematis recta L. var, paniculata Thunb, Senninso 13 oll Carpesium abrotanoides L, Yabutabako 13 sit Eupatorium japonicum Thunb, Sawa hiyodori 12 1.0 Acanthopanax pivaricatum §S, et Z. Ukogi 12 1.0 Arundinella anomala Steud. Todashiba 12 1.0 Gentiana scabra Bge. var Buergeri Max. Rind6 12 1.0 Senecio krameri Fr. et Sav. Yaburegasa 12 1.0 Pieris aquilina L, Warabi 12 1.0 Sanguisorba efficinalis L. Waremok6 12 1.0 Plectranthus graucocalyx Max, var japonicus Max, Hikiokoshi if) 0.9 Saussurea Tanakae Fy, et. Sav. var. phyllolepis Max. | ‘Tohiren 8 \ Serratula atriplicifolia B. et 11. Kumotorihokuchi ) Manettia iginta (Vell.) Kwaens6 8 Atractylodes lyrata S. et Z. Okera il Aster indicus L, Yomena 6 a2 Patrinia villosa Juss. Otokoeshi 5 Brachypodium silvaticum R, et S. Yamakamoji gusa 2 Osmunda regalis L, var japonica Milde, Zenmai 3 1170 100 ii Ueber die Einwirkung des H7/ava-Brennens. Omiyama (Siidseite). (fiir 3 Jahre nicht gebrannt) (Neigung = 34°) (Datum 11/4 u. 26/4) lateinischer Name, Miscanthus sinensis Anders. Chrysanthemum sinense var. japanicum Max. Disporum sesile Don, Carex breviculmis R. Br. Agrimonia viscidula Bge, var, japonica Miq. Houttuynia cordata Thunb. Plectranthus inconspicus Miq. Cirsium spicatum Maxim, Thalictrum minus L, var, elatum Lecoy, Senecio Krameri Fr. et Sav, Artemisia vulgaris L, var. indica Max. Athyrium nipponicum Bak, Carex lanceolata Boo't. Oxalis corniculata L. Poa trivialis L, Dioscoria Tokoro Makino, Gentiana scabra Bge, var. Buergeri Max, Paederia tomentosa BI, Andropogon micranthus Kth, Dianthus superbus L, Crematis recta L, var, peniculata Thunb, Astilbe thunbergii Mig. Crepis japonica Benth. Lysimachia clethroides Duby. Lygodium japonicum Sw. Lactuca thanbergiana (A, Gr.) Maxim, japanischer Name. Susuki Ryunogiku Hochakus6 Aosuge Kinmizuhiki Dokudami Yamahakka Yamaazami Akikaramatsu Yaburegasa Yomogi TInuwarabi Hikasuge Katabami Hime ichigotsunagi Onidokoro Rindd Hekusokazura Hime aburasusuki Kawaranadeshiko Senninso ToriashishGma Onitabirako Oltatoranoo Tsurushinobu Nigana Zah! der Gr, in 2|_].m. 2 “I ~ to 2 Nv Co Ww ; eg tte tu ty H : ~ tu I] we 286 Q. Shishido: Aster japonicus Miq. Yamashirogiku 7) 1.4 Aster scaber Thunb. Shirayamagiku 16 Tes Pteris aquilina L, Warabi 16 192} Vitis thunbergii S, et Z. Ebizuru 15 1.3 Tricyrtis japon‘ca Miq. Tfototogisuso 15 1.3 Rubus incisus Thunb, [Nigaichigo] 15 1-2 Lespedeza pilosa S. et Z, Nekohagi 15 12 Brachypodium silvaticum R. ct S. Yamakamojigusa 15 eZ Potentilla fragarioides L, Kizimushiro 15 1.2 Smilacina japonica A, Gr. Yukizasa 14 2, Carpesium abrotanoides 1. Vabutabako 14 L2 Sanguisorba officinalis I, Waremok6G 13 vies Atractylodes lyrata S. et Z. Okera II 0.9 Patrinia scabiosaefolia Link Ominaeschi 8 Carex conica Boott. Hime kansuge 8 FEupatorium japonicum Thunb, Sawa hiyodori 6 = Saussurea Tanakae Fr, et Sav. var. phyllolepis Max. To hiren 3 1212 100 Es wird daraus klar, dass diese Hara (Omiyama) verhiltnissmissig viele Grasspecies enthalt; es sind hier die erwaihnten Lichtgriiser seltener, wahrend andere Arten in grésserer Zahl gefunden werden. Aus dem Aufwuchse kann die Bodenbeschaffenheit beurtheilt werden; es gedeihen auf schlechtem Boden eben schlechte Grasarten und auf gwtem Boden die besseren Arten. | Auch die gréssere Zahl der Gewichsarten deutet an, dass ein Boden nahrungsreicher ist; in nahrungsarmem Boden kénnen nur wenige Species fortkommen bei sonst gleichen Verhiltnissen. Man kann daher schliessen. dass die suerst genannte //ara (Kiriddshi) nahrungsirmer ist als die szvezte (Omiyama). I]. KAPITEL. Folgen des Brennens einer Hara. (A) Verdnderung der Gewichsarten durch das Brennen. Die Verinderung der Gewiichsspecies lisst sich aus Untersuch- ungen in Kiyosumi erkennen. Diese H/ava wurde, wie schon bemerkt, auf Ueber die Einwirkung des Ifava-Brennens. 287 verschiedenen Theilen in verschiedenen Zwischenriumen gebrannt. Die ae | folgenden Tabellen beleuchten in ihren Ergebnissen diese Frage (die einzelne Probefliche=20 LJ. m. gross.) Kajisaka (Nordseite). (aljihrlich gebrannt) (Neigung = 39°) (Probeflache=20 Li. m.) | (Datum 10/4 u. 26/4) lateinischer Name. japanischer Name. % Miscanthus sinensis Anders. Susuki 28 Sanguisorba officinalis L, Waremok6 s Carex Duvaliana Fr, et Sav. Kesuge S Brachypodium silvaticum Rk. et S. Yamakamajigusa 7 Pteris aquilina L. Warabi 5 Gerbera anandria Sch, Bip. Senbonyari 5 Thalictrum minus L, var. eltaum Lecoy, Akikaramatsu 5 Phegopteris Totta Mett, Mizoshida 5 Astilbe Thunbergii Miq. ToriashishGma | 4 Artemisia japonica Thunb. Otokoyomogi 4 Patrinia villosa Juss, Otokoeshi 3 Polygonum cuspidatum S. et Z. Itadori 3 Carex lanceolata Boott. Hikagesuge 2 Atractylades lyrata S, et Z. Okera 2 Centaurea atriplicifolia (DC,) Yamabokuchi 2 Senecio krameri Fr, et Sav. Yaburegasa I Aster scaber Thunb, Shirayamagiku I Euphorbia sieboldiana Mor, et Dee. Natsutidai I Aster trinervius Roxb, var, japonica Max, Konkiku Osmunda regalis L, var, japonica Milde. Zemmai | Angelica decursiva Mig. Nodake | Serratula coronata L, Cirsium spicatum Maxim, Tamaboki Yamaazami 288 QO. Shishido: Sanicula elata Miq. | Umanomitsuba Aconitum sinense S, et Z. Torikabuto Sedum kamtschachicum Fisch. Kirinso Disporium pullum Salish. Toéchikuran Cypripedilum japonicum Thunb, Kumagaiso 6. Saussurea ussuriensis Max, Kikuazami Coelopleurum Gmelini Ledeb. Shishiudo Adenophora verticillata Fisch. var, verticillata (Fr. et Sav.) | Tsurigane ninjin Saussurea affinis Spr. Kitsuneazami Petasites japonicus Miq. Fuki Hosta Sieboldiana Engl. TO gibdshi Solidago Virga-aurea L. Awadachiso Patrinia scabiosaefolia Link. Ominaeshi Leonurus macranthus Maxim, Kisewata Smilax china L, [Sankirai] 100 Kajisaka (Siidseite). (alhéhrlich gebrannt) (Neigung = 32°) (Probeflache = 20 L1. m.) (Datum 10/4 u. 26/4) lateinischer Name. japanischer Name. % Miscanthus sinensis Anders, Susuki 34 Brachypodium selvaticum R, et S. Yamakamojigusa 6 Arundinella anomala Steud. ‘Todashiba 5 Thalictrum minus L, var. elatum Locoy Akikaramatsu 5 Sanguisorba officinalis L, Waremoko6 4 Potentilla fragarioides L. var. ternata Maxim. Mitsubatsuchiguri 4 Plectlanthus glaucocalyx Max, var, japonicus Max, Hikiokoshi 3 Phegopteris Totta Mett, Mizoshida 3 Artemisia japonica Thunb, Otoko yomogi 3 Poa trivialis L, Ilime ichigotsunagi 3 Ueber die Einwirkung des Hara-Brennens, 289 Aster scaber Thunb, Shirayamagiku 3 Senecio krameri Fr, et Say, Yaburegasa 2 Euphorbia Sieboldiana Mor, et Dec, Natsutédai 2 Viola Patrinii DC. var, chinensis Ging, Sumire 2 Taraxacum officinale Wigg. var, glausescens Koch. Tanpopo | 2 Pteris aquilina 1, Warabi | 2 Lysimachia clethroides Duby, Okatoranoo | 2 Aster japonicus Miq. Yamashirogiku | I Disporum sesile Don, H6chakus6 | I Cirsium spicatum (Maxim.) Yama azami I Saussurea ussuriensis Maxim, Kiku_azami I Polygonum cuspidatum §, et Z, Itadori I Cozlopleurum Gmelini Ledeb, Shishiudo Seseli Libanostis Koch, var. daucifolia DE: Ibukibdfu Ixeris Thunbergii A, Gr, Nigana Picris hicracicides L, var, Japonica Rgl., Kozorina Viola japonica Longsd, Kosumire | Dioscorea Tokoro Makino, Onidokoro | Melandryum firmum Rohrb, Fushiguro Senecio campestris DC, Sawaoguruma Calystegia sepium R, Br, Hirugao Allium japonicum Rgl, Yamarakkyd Serratula corronata L, Tamuraso ey Se Agrostis perennans Tuck, Nukabo Lilium Maximowiezii Rel. Koniyuri Rumex acetosa L, Suiba Hotarukazura Lithospermum Zollingeri A. DC, Cryptogramme japonica Prantl, Tachishinobu Polygonatum giganteum Dietr, var, Thunbergii Max. Narukoyuri Inula salicina L, Kasensd Cimicifuga foetida L, var, simplex Huth, Sarashinashoma Vicia unijuga Al. Br. [Nantenhagi) Rubus parvifolius [Nawashiro ichigo) Graminea Sp, Graminea Sp. 290 0. Shishido: Omiyama (Nordseite), (seit 3 Fahre nicht gebrannt) . (Neigung = 34°) (Probeflache=2 (©. m.) (Datum 12/4 u. 26/4) lateinischer Name. Miscanthus sinensis Anders, Aster japonicus Miq. Carex breviculmis R, Br. Carex Duvariana Fr. et Sav. Arundinella anomala Steud. Disporum sesile Don. Serratula coronata L. Potentilla fragarioides L. var. Ternata Max. Arteneisia vulgaris L. var, indica Max. Kraunhia floribunda (willd) Taub. Picris hicracioides L, var, japonicayRgl. Adenophora verticillata Fisch. var.|verticillata (Ir. et Sav.) Gentiana scabra Bge, var. Buergeri Max. Artemisia japonica Thunb, Agrimonia viscidula Bg, var, japonica Miq. Euphorbia Sieboldiana Mor, et Dec. Pteris aquilina L, Senecio Krameri Ir, et Sav. Chrysanthemum sinense Sab, var, japonicum Max. Aster scaber Thunb. Angelica decursiva Miq. Clematis recta L, var, paniculata (‘Thunb.) Brachypodium silvaticum R., et S. Patrinia villosa Juss. Rumex acetosa L, japanischer Name. Susuki Yamashirogiku Aosuge Kesuge Todashiba Hochakuso Tamaboki Mitsubatsuchiguri z ; Yomogi Fuji Kozorina Tsuriganeninjin Rinds Otokoyomogi Kinmizuhiki Natsutodai Warabi Yaburegasa Ryanogiku Shirayamagiku Nodake Senninso Yamakamojigusa Otokoeshi Suiba G2 Einleitung. 261 Brachypodium japonicum Miq. Kamojigusa | Solidago virga-aurea L. Awadachiso | Cirsium spicatum Maxim. ; Yamaazami | Calanthe discolor Lind]. Ebine Saussurea ussuriensis Maxim. Kikvazami | Petasites japonicus Miq. Fuki | Coclopleurum gmelini Ledeb. Shishiudo Phytolocea acinosa Roxb, var, esculenta Max, Yamagobo | Penthorum sedoides L. var, chinense Max. Sawa shion | Viola silvestris Kit, var. grypoceras A, Gr. Tachitsubosumire Astilbe Thunbergii Miq. ToriashishGma ‘a Osmunda regalis L. var. japonica Milde. Zemmai Dioscorea tokoro Makino, Onidokoro Lysimachia clethroides Duby. Okatoranoo Polygonum cuspidatum S, et Z. Itadori | Viburnum dilatatum Thunb, {Gamazum‘] Quercus glandulifera BI. [Konara] | Quercus serrata Thunb. [Kunugi] | Rhododendron indicum Sw, var, Kaempferi Max. [Yamatsutsuji] Rubus parvifolius L, | [Nawashiroichigo] | Smilax china L. | [Sankirai] | Lespedeza bicolor Turcz. | [Hagi] Spiraea japonica L. f. | [Shimotsuke] Omiyama (Siidseite). (seit 3 Jahren nicht gebrannt) (Neigung = 34°) (Probefliche =2 L1. m.) (Datum 10/4 u. 26/4) lateinischer Name, japanischer Name, Miscanthus sinensis Anders. Susuki Phegopteris totta Mett, Mizoshida 292 Aspidium aristatus Sw. Carex conica Booit. Serratula coronata L. Disporum sessile Don, Agrimonia pilosa Ledeb, Chrysanthemum sinense Sab, var. japonicum Max. Carex Morrowi Boott. Houttuynia cordata Thunb. Potentilla fragarioides L, var. ternata Max. Picris hicracioides L, var, japonica Rgl. Viola silvestris Kit. var. grypoceras A. Gr. Artemisia japonica Thunb, Angelica decursiva Miq. Arundinella anomata Steud. Sodum kamtschoticum Fisch, Carex duvariana Fr, et Sav. Aspidium lepidocaulon Hook, Eupatorium Kirilowii Turez. Brachypodium silvaticum R, et S, Aster scaber Thunb, Angelica polymorpha Maxim, Euphorbia sieboldiana Mor, et Dec. Lactuca Thunbergiana (A. Gr.) Maxim. Gentiana scabra Bge, var, Buerger! Max. Pteris aquilina L, Senecio crameri Fr, et Sav. Osmunda regalis L. var, japonica Mild. Athyrium filix femina Roth, Artemisia vulgaris L, var. indica Maxim. Cryptogramme japonica Prantl. Patrinia scabiosaefolia Link, Patrinia villosa Juss. Ophiopogon japonicus Ker, Dianthus superbus L, 0. Shishido: Hosoba kamawarabi Himekansuge Tamaboki Hochakuso Kinmizuhiki Ryunogiku Kansuge Dokudami Mitsubatsuchiguri Kozorina Tachitsubosumire Otokoyomogi Nodake Todashiba Kirins6 Kesuge Orizurushida Sawahiyodori Yamakamojigusa Shirayamagiku Shiranesenkiu Natsutodai Nigana Rindo Warabi Yaburegasa Zemmai Meshida Yomogi Tachishinobu Ominaeshi Otokoeshi Yomohige Kawaranadeschiko Oo Ww WO W] W Einleitung. 29 Saussurea ussuriensis Maxim. Kikuazami Erigeron annuus Pers, Himejoon Lonicera japonica Thunb. Nindé Cypripedium japonicum Thunb, Kumagais6 Plantago major L, var, asiatica Dene. Obako Crematis recta L, var. peniculata Thunb. Sennins6 Tris japonica Thunb, Shoga Petasites japonicus Miq. Fuki 17. Lysimachia ceiroides Duby. Okatoranoo Arisaema japonicum BI. Tennansho Vitis Thunbergii S. et Z, Ebizuru Coelopeurum gmelini Ledeb, Shishiudo Clematis japonica Thunb. Hanshézuru Rubus parvifolius L, Akebia quinata Dene. Rosa Wichuraiana Crep, Akebia lobata Dene. Polygara japonica Hautt, Deutzia gracilis S, et Z. Spiraea Thunbergii Sieb. Smilax china L, [Nawashiroichigo] [Akebi] [ Terihanoibara} [Mitsubaakebi] [Himehagi] [Himeutsugi] [Iwayanagi] [Sankirai] Acanthopanax ricinifolium S. et Z. [Bodara] Quercus glandulifera Bl. [Konara] Quercus serrata Thunb, {[Kunugi] SSS Omiyama (Siidseite). (seit 6 Jahre nicht gebrannt) (Neigung = 36°) (Probefliche=2 0. m.) (Datum 12/4 u. 26/4) lateinischer Name, japanischer Name, % EE ee ee, ee ee fe Miscanthus sinensis Anders, Susuki 20 294 ©. Shishido: Carex duvariana Fr, et Sar. Poa trivialis L. Cryptogramme japonica Prantl. Viola silvestris Kit. var. grypoceras A, Gr. Carex confertiflora Boott. Trycyrtis chirta Hook. Polygonatum giganteum Dietr, var, Thunbergii Max, Phegopteris Totta Mett. Eupatorium japonicum Thunb, Thalictrum minus L, var, elatum Lecoy. Gentiana Zollingeri Fawe. Gentiana scabra Bge. var. Buergeri Max, Hypericum sampsoni Hee, Salvia japonica Thunb. var, bipinnata Fr, et Sav. Brachypodium silvaticum R. et S. Potentilla fragarioides L. var, ternata Max. Aster japonicus Miq. Rubia cordifolia L. var, mungista Miq. Gynostemma pedata Bl, Pteris aquilina L, Ophiopogon japonicus Ker, Aconitum fischeri Reich. Artemisia vulgaris L, var, indica Maxim. Patrinia scabiosaefolia Link. Patrinia villosa Juss. Polygonum cuspidatum S, et Z. Arisaema japonicum Bl], Poa acroleuca Stena. Dianthus superbus L. Osmorhiza japonica S, et Z. Senecio Krameri Fr, et Say. Cirsium spicatum Maxim. Viola japonica Langsd. Clematis recta L, var. paniculata Thunb. Kesuge Hime ichigotsunagi Tachishinobu Tachitsubosumire Shirasuge Hototogisuso Narukoyuri Mizoshida Hiyodoribana Akikaramatsu Fuderinds Rindo Tsukinukiotogiri Akinotamuraso Yamakamojigusa Mitsubatsuchiguri Yamashirogiku Akane Amachazuru Warabi Janohige Torikabuto Yomogi Ominaeshi Otokoeshi Ttadori Tennanshd Mizo ichigotsunagi Kawara nadeshiko Yabuninjin Yaburegasa Yamaazami Kosumire Sennins6 wo -»- PP fh nO NWN oP) N Einleitung. Asarum caulescens Miq. Aster scaber Thunb. Plectranthus inflexus Vahl. Clematis apiifolia DC, Aspiduim sculeatum Doell, var. japonicum (Fr. et Say.) Taraxaogum officinale Wigg. var. glauceescens Koch, Lonicera japonica Thunb, Petasites japonicus Miq. Platanthera chlorantha Cust. Inula salicina L. Saussurea ussurriensis Maxim, Bromus japonicus Thunb, Astilbe Thunbergii Miq. Calanthe reflexa Maxim, Hedera helix L, Rubus Buergeri Miq. Viburnmu dilatatum Thunb. Akebia quinata Dene. Akebia lobata Dene. Euonymus alata K. Kock. var. subtriflora Fr. et Sav. Quercus glandulifera Bl. Rubus palmatus Thunb. Aucuba japonica Thunb, Torreya nucifera S. et Z. Quercus myrsinaefolia 131, Spiraea japonica L, f. Lespedeza bicolor Turcz, Smilax china L, Vaccinium bracteatum Thunb, Deutzia gracilis S. et 7. Litsea glauca Sieb, Rhododendron indicum Sw, var, Kaempferi Max, Lindera selicea BI. Clethra barvinervis S. et Z. Diervilla grandiflora S, et Z, Kan aoi Shirayamagiku Yama hakka Botanzuru Inode Tanpopo Nindo Fuki GinbaisG Kasenso Kikuazami Suzumenochahiki ToriashishOma Natsuebine [Fuyuzuta] [Fuyuichigo] [Gamazumi] [Akebi] [Mitsubaakebi] [Komayumi] [Konara] [Kiichigo] {Aoki} [Kaya] [Shirakashi] [Shimotsuke] (Hagi] [Sankirai] (Shashanpo] (Himeutsugi] [Shirodamo] [Yamatsutsuji] [Kuromoji} [RyS6bu] (Hakoneutsugi] 206 0. Shishido: Musadogadai (Siidseite). (seit 6 Jahre nicht gebrannt) (Neigung = 29°) (Probeflache =20 L1: m.) (Datum 11/4 u. 27/4) lateinischer Name. Miscanthus sinensis Anders. Carex breviculmis R. Br. Carex duvariana Fr, et Sav. Viola silvestris Kit. var. grypoceras A. Gr, Artemisia vulgaris L, var, indica Maxim, Angelica anomala Pall. Aster trinervius Roxb. var. adustus Maxim. Disporum sessile Don, Viola patrinii var, chinensis Ging. Brachypodium japonicum Miq. Aspidium dissectum Mett. Houttuynia cordata Thunb. Thalictrum minus L, var. elatum Lecoy. Patrinia villosa Juss. Lysimachia clethroides Duby. Aster japonicus Miq. Viola japonica Langsd. Aster scaber Thunb. Pteris aquilina L. Arimonia viscidula Bge. var. japonica’ Miq. Cirsium spicatum Maxim. Rubia cordifolia L, var, mungista Miq. Plectranthus glaucocalyx Max, var, japonicus Max, Galium asprellum Michx. Senecio Krameri Fr, et Sav, japanischer Name. Susuki Aosuge Kesuge Tachitsubosumire Yomogi Yoroizasa Konkiku Hochakus6 Sumire Kamojigusa Hoschida Dokudami Akikaramatsu Otokoeschi Okutoranoo Yamashirogiku Kosumire Schirayamagiku Warabi Kinmizuhiki Yamaazami Akane Hikiokoschi Obayaemugura Yaburegasa Einleitung. Scrophularia patriniana Wydl. Euphorbia sieboldiana Mor, et Dic. Vitis Thunbergii S, et Z. Serratula coronata L, Plantago major L. var. asiatica Dene. Belamacanda chinensis Lem, Potentilla fragarioides L, var, ternata Maxim. Galanium nepalense Sweet. Dianthus superbus L, Lactuca Thunbergiana (A. Gr.) Maxim. Angelica decurtiva Miq. Petasites japonicus Miq. Clematis recta L. var, paniculata Thunb. Osmunda regalis L, var. japonica Milde. Platanthera mandarinorum Reich, f, Cirsium joponicum DC, Akebia quinata Dene. Clematis apiifolia DC. Rosa multiflora Thunb. Polygala japonica Hautt. Rubus parvifolius L, Akebia lobata Dene. Lespedeza bicolor Turcz. Deutzia gracilis S, et Z. Quercus glandulifera Bl. Smilax china L, Rubus palmatus Thunb, Spiraea japonica L, f, Kraunhia floribunda (Willd) Taub. Viburnum dilatatum Thunb, Diervilla grandiflora S, et Z. Spiraea Thunbergii Sieb, 207 Hinano usutsubo I Natsutodai | I Ebizuru I Tamuraso Obako Hidgi | Mitsubatsuchiguri Furosd Kawara nadeshiko Nigana Nodake Fuki Sennins6 Zemmai Yamasagis6 Noazami [Akebi] Botanzuru to WM [Noibara] (Himehagi] [Nawashiroichigo] [Mitsubaake bi] (Hagi] (Himeutsugi] [Konara] [Sankirai] [Kiichigo] [Shimotsuke] [Fuji] [Gamazumi] [Hakoncutsugi] [Iwayanagi] _— Suzuriischi ©. Shishido: (Siidseite) (liber 8 Jahre nicht gebrannt) (Neigung = 33°) (Probefliche = 20 LI. m.) (Datum 12/4 u. 27/4) lateinischer Name. Miscanthus sinensis Anders, Carex lanceolata Boott. Phegopteris Totta Mett. Viola silvestris Kit. var. grypoceras A. Gr, Dianthus superbus L. Ixeris Thunbergii A. Gr, Aspidium aristatum Sw. Artemisia japonica Thunb. Tricytis hirta Hook. Muhlenbergia Huegerii Trin. Liliam Maximowicizii Regel. Sedum Kamtschaticum Fisch. Viola Patrinii DC. var. chinensis Ging. Carpesium cernuum I. Brachypodium silvaticum R, et S. Woodwardia radicans Sm, var, orientalis}]irs, Osmunda regalis L. var. japonica Mild. Potentilla fragarioides L. Pteris cretica L. Rubia cordifolia L. var, mungista Miq. Euphorbia Sieboldiana Morr, et Dene. Cirsium spicatum (Maxim) Pteris aquilina L. Aster japonicus Miq. Gentiana scabra Bge. var. Buergeri Maxim, japonischer Name. Susuki Hikagesuge Mizoshida Tachitsubosumire Kawaranadeshiko Nigana Hosoba kanawarabi Otokoyomogi Hototogisusd Onezumigaya KOniyuri Kirinso Sumire Sajigankubiso Yamakamojigusa Komochishida Zemmai Kijimushiro Obainomotoso Akane Natsutddai Yamaazami Warabi Yamashirogiku Rindo G2 oP) | | Ueber die Einwirkung des Hara-Brennens. Chrysanthemum sinense Sab, var. japonicum Max. Arimonia cordifolia L. var. mungista Miq. Artemisia vulgaris L. var, indica Maxim. Solidago Virga-aurea L. Senecio krameri Fr. et Sav. Saussurea japonica DC, Seseli Libanostis Koch, var. daucifolia DC. Serratula coronata L. Houttuynia cordata Thunb, Clematis japonica Thunb. Astilbe Thunbergii Miq. Polygonatum giganteum Dietr. var. Thunbergii Max. Petacites japonicus Miq. Thalictrum minus L, var, elatum Lecoy, ' Cymbidium virens Lindl. Lonicera japonica Thunb. Inula salicina L, Atractylis lancea Thunb, Saussurea ussuriensis Maxim, Rhodea japonica Rhot. Ligularia Keempferi S, et Z, Rubus parvifolius L, Polygara japonica Hautt, Rubus incisus Thunb. Hedera helix L, var, colchico C, Koch, Akebia lobata Dene. Rubus palmatus Thunb, Quercus glandulifera Bl, Deutzia gracilis S, et Z. Deutzia scabra Thunb, Diervilla’ grandiflora S, et Z, Berberis Thunbergii DC, Lindera selicea Bl. Rhododendron indicum Sw, var, Keempferi Max, Ryundgiku Kinmizuhiki Yomogi Akinokirins3 Yaburegasa Himehigotai Ibukibdfu Tamurasd Dokudami Hanshozuru ToriashishOma Narukoyuri Fuki Akikaramatsu Shunran Suikazura Kasenso Okera Kikuazami Omoto Tsuwabuki [Nawashiroichigo] [Himehagi] [Nigaichigo] [Fuyuichigo] [Mitsuba-akebi] [Momijiichigo] [Konara] ([Himeutsugi] [Utsugi] [Hakoneutsugi] [Megi] [Kuromoji} [Yamatsutsuji] to oO \O (oe) 0. Shishido: Smilax china L. [Sankirai] Eurya japonica Thunb. | [Hisakaki] Lespedeza bicolor Turcz. [Hagi] Clethra barvinervis S. et Z. [Rydbu] Litsea glauca Sieb. [Shirodamo] Spiraca japonica L., f. [Shimotsuke] Aucuba japonica Thunb. ~ [Aoki] 100 Suzuriishi (Nordseite). (bis 24 Meiji (1891) alljahrlich und spater im Jahre 33 Meiji (1900) gebrannt) (Neigung = 32°) (Probeilache =20 Ll im) (Datum 13/4 u. 27/4) lateinischer Name. | japanischer Name, % Miscanthus sinensis Anders. Susuki 12 Phegopteris totta Mett. Mizoshida 5 Chrysanthemum sinense Sab. var. japonicum Max, Ryunégiku 4 Gentiana scabra Bge, var. Buergeri Maxim. Rindo 4 Carex duvariana Fr, et Sav. Kesuge g Carex confertiflora Booit. Shirasuge 3 Brachypodium silvaticum R. et 5. Yamakamojigusa 2 Carex brunnea Thunb. Nakirisuge = Cirsium spicatum Maxim, Yamaazami 3 Kraunhia floribunda (Willd) Taub. Fuji s Disporum sessile Don, Hochakus6 2 Trityrtis hirta Hook, Hototogisus6 2 Carpesium cernuum L, Sajigankubiso 2 Clematis heracleifolia DC. var, stans S. et Z. Kkusabotan 2 Lactuca denticulata Maxim. Yakushiso 2 Patrinia villosa Juss. Otokoeshi 2 — ae Ueber die Einwirkung des Hlava-Brennens. ‘Thalictrum minus L, var. elatum Lecoy. Campanula punctata Lam, Solidago Virga-aurea L, Lysimachia clethroides Duby. Patrinia scabiosaefolia Link, Dianthus superbus L. Osmunda regalis L, var, japonica Milde. Picris hieracioides L, var. japonica Rel. Aristolochia Keempferi Willd. Serratula coronata L. Heteropappus hispidus Less, Cryptogramme japonica Prantl, Eupatorium japonicum Thunb, Inula salicina L, Woodwardia radicans Sm, var. orientalis Lirs. Asteromeea indica BI, Clematis recta L. var. paniculata Thunb, Petasites japonicus Miq. Aster scaber Thunb. Senecio Krameri Fr. et Sav. Potentilla fragarioides L, var, ternata Maxim, Houttuynia cordata Thunb. F Coelopleurum gmelini Ledeb, Astilbe Thunbergii Mig. Euphorbia sieboldiana Mor,et Dic, Artemisia japonica Thunb. Seseli Libanostis Koch, var. daucifolia DC. Plantago major L, var, asiatica Dene. Ligularia Keempferi S, et Z, Cymbidium virens Lindl. _ Allium japonicum Rgl. Lonicera japonica Thunb, Rubus Buergeri Miq. Spirsea japonica L, f. Okikaramatsu Hotarubukuro Akinakirins6 Okatoranoo Ominaeshi Kawaranadeshiko Zemmai K6zorina Oba umanosuzukusa Tamuraso Yamajinogiku Tachishinobu Hiyodoribana Kasenso Komochishida Yomena Senninso Fuki Shirayamagiku Yaburegasa Mitsubatsuchiguri Dokudami Shishiudo Toriaskish6ma Natsutédai © Otokoyomogi Ibukibéfu Obako Tsuwabuki Shunran Yamarakkyod Nind6 [Feyuichigo} [Nawashiro'chigo] Ww lo 301 302 @. Shishido: Rubus parvifolius L. ' [Shimotsuke] Polygara japonica Hautt. Himehagi Akebia lobata Decne. Mitsuba-akebi Ficus foveolata Wall. Ttabikazura 25 Quercus glandulifera Bl. Konara Quercus serrata Thunb. Kunugi Smilax china L, Sankirai Rubus palmatus Thunb. Kiichigo Clethra barvinernis S, et Z, Ryobu Diervilla grandiflora S, ct Z. Hakoneutsugi Deutzia gracilis S, et Z. Himeuisugi Deutzia scabra Thunb. Utsugi Lindera selicera bl, Kuromoji Torreya nucifera S, et Z. Kaya Aucuba japonica Thunb. Aoki Eurya japonica Thenb Hisakaki Spireea Thunbergii Sieb. Iwayanagi Z Ardisia crenata Sims. Manryo Rhododendron indicum Sw, var. Keempferi Max. Yamatsutsuji Macleya cordata R. Br. Chanpagiku Poa trivialis L, Himeichigotsunagi Pteris serrulata L, f. Tnomotos6 Pteris aquilina L. Warabi | Sannodai (Nordseite). (bis 24 Meiji (1891) alljahrlich und im Jahre 33 Meiji (1900) gebrannt) (Neigung = 35°) (Probefliche=20 LI. m.) Datum 14/4 u. 27/4) lateinischer Name, | japanischer Name, | % | | Miscanthus sinensis Anders, | Susuki | 16 ; Ueber die Einwirkung des 7iava-Brennens. Carex breviculmis R. Br, Picris hicracioides L. var, japonica Rgl., Carex brunnea Thunb, Viola silvestris Kit. var, grypoceras A, Gr, Tricyrtis hirta Hook. Gentiana scabra Bge, var. Buergeri Maxim, Salvia japonica Thunb, var, bipinnata Fr, et Sav. Carpesium cernuum L, Artemisia vulgaris 1. var, indisa Maxim, Aster japonicus Miq. Crawfurdia pterygocalyx Hemsl. Carex conica Boott. Polygonatum lasianthum Maxim. Lactuca danticulata Maxim, Aspidium lacerum Sw. Asarum Blumei Duch. Eupatorium japonicum Thunb, Aspidium crythrosarum Eat. Aristolochia Keempferi Willd. Osmunda regalis L.. var, japonica Mild. Patrinia villosa Juss. Astilba Thunbergii Miq. Cirsium spicatum Max. Senecio Krameri Fr. et Sav. Cypripedilum japonicum Thunb, Cinnbidium virens Lindl, Patrinia scabiosaefolia Link, Clematis japonica Thunb, Lilium auratum Lindl, Houttuynia cordata Thunb. Pteris aquilina L, Brachypodium silvaticum R, et S. Liguralia Keempferi S, et Z. Rhodea japonica Rhot, Aosuge K6zorina Nakirisuge Tachitsubosumire Hototogisusé Rindo Akinotamuraso Sajigankubiso Yomogi Yamashirogiku Tsururind6d Himekansvuge Miyamanarukoyuri Yakushiso Kumawarabi Kan-aoi Hiyodoribana Benishida Oba-umanosuzukusa Zemmai Otokoeshi Toriashish6ma Yamaazami Yaburegasa Kumagaiso Shunran Ominsacsni * PRanshoézurn Yamayuri Dokudami Warabi Yamakamojigusa Tsuwabuki Omoto O04 Clematis recta J., var, paniculata (Thunb). Mitchella undulata S. et Z. Lindera umbellata Thunb. Quercus glandulifera Bl. Rhus succedanea L. Rosa Wichuraiana Crep. Hedera belix L. var. colchica C. Kech. Akebia quinata Dene. Spiraea japonica L. f. Trachelospermum jasminoides Lemaire. Ficus foveolata Wall. Rubus Buergeri Miq. Akebia lobata Dene. Vaccinium bracteatum Thunb. Pieris japonica D, Don. Illicium anisotum L. Thea japonica (L.) Wois. Eurya japonica Thunb, Rubus palmatus Thunb. Rosa multiflora Thunb. 0. Shishido: Rhododendron indicum Sw. var, Kzempferi Max. Osmanthus aquifolium B, et H. Lindela selicea BI. Diervilla grandiflora S, et Z. Deutzea gracilis S. et Z, Rubus parvifolius L. Smilax china L, Litsea glauca Sieh. Zanthoxylum schinnifolium S. et Z. Senninso Tsuruaridoshi [Kanakuginoki] [Konara] [Tsutaurushi] [Terihanoibara] [Fuyuzuta] [Akebi] [Shimotsuke] [Teikakazura ] [Itabikazura] [Fuyuichigo] [Mitsubaakebi] J 2 ios) [Shashanpo] [Asebi] [Shikimi] Tsubaki Hisakaki Kiichigo Noibara Yamatsutsuji Hiiragi Kuromoji Hakoneutsugi Himeutsugi Shimotsuke Sankirai Shirodamo Inusansh6d Ueber ice Einwirkung des lara-Brennens. Sannodai (Sidseite). (bis 24 Meiji (1891) alljahrlich und im Jahre 33 Meiji (1900) gebrannt) (Neigung = 31°) (Probeflache = 20 (1. m.) (Datum 14/4 u. 27/4) lateinischer Name. Miscanthus sinensis Anders. Oxalis corniculata L. Carex brevicurmis R. Br. Viola Keiskei Miq. Carex confertiflora Bott, Phagopteris totta Mett. Disporum sessele Don. Carex brunnea Thunb, Salvia japonica Thunb. var. Buergeri Max. Brachypodium silvaticum R, et S. Viola silvestris Kit, var. grypoceras A, Gr, Carpesium abrotanoides L, Eupatorium japonicum Thunb. Crepis japonica Benth, Bothriospermum tenellum Fisch et Mey, var, aspergoides Max) Aster japonicus Miq. Eupatorium Kirilowii ‘Turcz. Cirsium spicatum Max, Euphorbia sieboldiana Mor, et Dec. Aspidium aristatum Sw. Peracarpa circeeoides H, Fee, Artemisia vulgaris L. var, indica Maxim, Senecio Krameri Fr, et Sav. Lactuca debilis (Thunb), Maxim, japanischer Name, Susuki Katabami Aosuge Marubasumire Shirasuge Mizoshida Hochakus6 Nakirisuge Akinotamuras6 Yamakamojigusa Tachitsubosumire Yabutabako Hiyodoribana Onitabirako Hanaibana Yamashirogiku Sawahiyodori Yamaazami Natsutodai Hosoba-kanawarabi Tanigikyd Yomogi Yaburegasa Jishibari ty Ww 306 0. Shishido: Pteris aquilina L. Asplenium liniolatam Thunb. Cypripidilum japonicum Thunb. Gnaphalium multiceps Wall. Rubia cordifolia L. var, Mungista Miq. Thalictrum minus L. var, elatum Lecoy. Cryptogramme japonica Prantl. Cirsium japonicum DC, Petasites japonicus Miq. Patrinia villosa Juss, Astilbe Thunbergii Miq. Cremasira Wallichiana Lind]. Gentiana scabra Bge, var, Buergiri Maxim, Lactuca denticulata Maxim, Cymbidium virens Lindl, Rhodea japonica Rhot, Clematis apéifolia’ DC, Akebia quinata Dene. Rubus incisus Thunb, Duchesnea indica Fock. Clematis japonica Thunb, Sambucus racemosa L, Myrica rubra S. et Z. Eurya japonica Thunb, Rhododendron indicum Sw, var, Kzempferi Max. Torreya nucifera S, et Z, Pieris japonica D, Don, Deutzia gracilis S, et Z. Lindera selicea Bl. Quercus glandulifera BI, Thea japonica (L,) Nois, Machilus Thunbergii S. et Z. Rosa multiflora Thunb. Acanthopanax ricinifolium S, et Z, Warabi Herashida Kumagaiso Chichikogusa Akane Akikaramatsu Tachishinobu Noazami Fuki Otokoeshi ToriashishOma Saihairan . Rindo Yakushiso Shunran Omoto Botanzuru [Akebi] [Nigaichigo] [Hebiichigo] [Hanshoézuru] [Niwatoko] [Yamamomo] [Hisakaki] [Yamatsutsuji] [Kaya] [Asebi] Himeutsugi Kuromoji Konara Tsubaki Tabu Noibara Bodara 1 f : II Ueber die Einwirkung des Hara-Brennens, 307 Litsea glauca Sieb, Shirodamo Rubus palmatus Thunb, . Kiichigo Diervilla grandiflora S, et Z, Hakoneutsugi Clematis recta L, var, as‘atica Dene, Sennins6 Lonicera japonica Thunb, Nind6 Nanamagari. (seit to Jahren nicht gebrannt) (Neigung = 35°) (Probefliche =20 U1. m.) (Datum 15/4 u. 26/4) lateinischer Name. SS ee ee eee japanischer Name. tg Miscanthus sinensis Anders, Susuki II Artemisia japonica Thunb, Otokoyomogi 5 Palygonatum officinale All, Amadokoro 4 Carex incisa Boott, Kawarasuge 4 Carex duvariana Fr, et Sav, Kesuge 4 Cryptogramme japonica Prantl, Tachishinobu 3 Picris hieratioides L, var, japonica Rel, Kozorina 3 Chrysanthemum sinense Sab, var, japonicum Max, Ryundgiku = | Viola japonica Langsd. Kosumire “ Aristolachia Kaempferi Willd, Oba-umanosusukusa 3 Disporum sesile Don, Hochakuso 2 Carex conica Boott, Himekansuge 2 Brachypodium silvaticum R. et S, Yamakamojigusa 2 Serratula coronata L, Tamurasd 2 Aster japonicus Miq, Yamashirogiku | 2 Thalictrum minus L, var, elatum Lecoy, Akikaramatsu 2 Potentilla fragarioides L, Kijimushiro 2 Polygonatum giganteum Dietr, var, Thunbergii Max, Narukoyuri | 2 Sedum kamtschaticum Fisch, 2 Kirinso | 308 ®. Shishido: Pteris aquilina L, Angelica decursiva Miq. Cirstum spicatum Maxim, Euphorbia Sieboldiana Mor, et Dec. Astilba Thunbergii Miq. Clematis recta L. var, paniculata Thunb, Patrinia villosa Juss. Osmunda regalis L, var. japonica Milde. Potentilla fragarioides L. var. ternata Maxim. Artemisia vulgaris L, var, indica Max, Saussurea ussuriensis Max, Senecio Krameri Fr, et Sav. Arisaema japonicum BI, Lithospermum zollingeri A, DC, Asarum Blumei Duch, Cimicifuga foetida L. var. simplex Huth, Lonicera japonica Thunb, Petasites japonicus Miq. Salvia japonica Thunb. var. bipinnata Fr, et Sav, Dianthus superbus L, Lilium auratum Lindl, Gentiana scabra Bge. var, Buergeri Max, Luzula campestris DC, var, capitata Miq. Plectranthus glaucocalyx Max, var. japonicus Max, Rubus palmatus Thunb, Rubus parvifolius L. Akebia lobata Dene, Vicia unijuga Al, Br. Enonymus alata K, Koch, var. subtriflora Fr, et Sav. Spiraea japonica L., f, Quercus glandulifera BI, Eurya japonica Thunb. Diervilla grandiflora S, et Z. Viburnum dilatatum Thunb, Warabi Nodake Yamaazami Natsutddai Toriashishoma Senninso Otokoeshi Zemmai Mitsubatsuchiguri Yomogt Kikuazami Yaburegasa Tennanshd Hotarukazura Kan-aoi Sarashinash6ma Ninds Fuki Akinotamuras6 Kawaranadeshiko Yamayuri Rindo Suzumenohiye Yamahakka [ Momijibaichigo] [Nawashiroichigo] [Mitsubaakebi] [Nantenhagi] [Kkomayumi] [Shimotsuke] [Konara] [Hisakaki] [Iakoneutsugi] [Gamazumi] 30 | | | Ueber die Einwirkang des Mava-Brennens. 309 Machilus Thunbergii 5, et Z. [Tabu] Clethra barvinervis S, et Z. [Ryobu] Smilax china L, [Sankirai] Quercus acuta Thunb. [Akagashi] Quercus myrsinaefolia BI, [Urajirogashi] Rosa multiflora Thunb. [Noibara] Rhododendron indicum Sw, var, Keempferi Max. [Yamatsutsuji] Rubus palmatus Thunb, [Kiichigo] Lindera selicea Bl, [Kuromoji] Spireea Thunbergii Sieb. [Iwayanagi] Musadogadai (Nordseite). (iiber 10 Jahre nicht gebrannt) (Neigung = 32°) (Probeflache = 20 WU). m.) (Datum 13/4 u. 27/4) lateinischer Name, japanischer Name, % Miscanthus sinensis Anders, Carex Ringoldiana Boott. Pertya Scandens Sch, Bip, var, ovata Maxim, Brachypedium silvaticum R, et S, Phegopteris Totta Mett. Carex duvariana Fr, et Sav. Viola silvestris Kit, var, grapoceras A, Gr, Carex brunnea Thunb, Chrysanthemum sinense Sab, var, japonicum Max, Aster trinervius Roxb, var, adustus Max, Tricyrtis hirta Hook, Saussurea japonica DC, Viola Patrinii DC, var, chinensis Ging, Ixeris Thunbergii A, Gr, Susuki Juzusuge Koyaboki Yamakamojigusa Mizoshida Kesuge Tachitsubosumire Nokirisuge Ryunogiku Kongiku Hototogisuss Himehigotai Sumire Nigana 310 Arimonia cordifolia L, ae mungista Miq. Salvia japonica Thunb, var. bipinnata Fr, et Sav. Aristolochia Keempferi Willd. Gentiana scabra Bge. var, Buergeri Maxim, Angelica decursiva Miq. Cirsium spicatum Maxim, Woodwardia radicans Sw, var. japonica Lirs, Polygonum cuspidatum 5. et Z. Aster japonicus Miq. Senecio Krameri Fr, et Sav. Cirsium japonicum DC, Potentilla fragarioides L, var. ternata Maxim, Astilbe Thunbergii Miq. Clematis recta L, var, paniculata Thunb, Thalictrum minus L, var, elatum Lecoy. Artemisia vulgaris L. var, indica Maxim, Lonicera japonica Thunb. Plantago major L, var. asiatica Dene. Lysimachia clethroides Duby. Lilium auratum Lindl. Liguleria Keempferi S, et Z. Belamacanda chinensis Lem. Calanthe discolor, Lind]. Clematis heracleifolia DC. Saussurea ussuriensis Maxim, Houttuynia cordata Thunb. Clematis heracleifolia DC. var. Stans (S. et Z.) Polygonatum giganteum Dietr, var, Thunbergii Max, Petasites japonicus Miq. Taraxacum officinale Wigg. var. glauciscens, Koch, Hedera helix L, var, colchica C, Koch. Akebia lobata Dene. Rubus parvifolius i, Rubus Buergeri Miq. @. Shishido: Kimmizuhiki Akirotamuraso Oba-umanosuzukusa Rindo Nodake Yamaazami Komochishida Itadori Yamashirogiku Yaburegasa Noazami Mitsubatsuchiguri ToriashishoOma Senninso Akikaramatsu Yomogi Nindo Obako Otokoeshi Yamayuri Tsuwabuki Hidgi Ebineran Tsuriganeso Kikuazami Dokudami Kusabotan Narukoyuri Fuki Tanpopo Tuyuzuta Mitsubaakebi Nawashiroich'go Fuyuichigo — ae Ueber die Einwirkung des Hara-Brennens. Clematis japonica Thunb, Acanthopanax spinosum Miq. Trachelospermum jasminoides Lemeire. Celastrus articulatus Thunb, Eurya japonica Thunb, @uercus myrsinaefolia BI, Quercus glandulifera BI. @uercus serrata Thunb. Torreya nucifera S, et Z. Viburnum dilatatum Thunb, Machilus Thunbergii S, et Z. Lindera selicea Bl, Deutzia gracilis S, et Z, Diervilla grandiflora S, et Z, Rosa multiflora Thunb, Rubus palmatus Tnunb, Eurya ochnaceae Szysz, Ardisia crenata Sims, Spiraea Thunbergii Sieb, Rhododendron indicum Sw, var, Kaempferi Max, Rubus parvifolius L, Aukuba japonica Thunb, Vaccinium bracteatum Thunb, Lespedeza bicolor Turcz, Hanshoézuru | Ukogi Teikakazura cA veo Tsuruumemodoki Hisakaki Urajirogashi Konara Kunugi Kaya Gamazumi Tabu Kuromoji Himeutsugi Hakoneutsugi Noibara Kiichigo Sakaki Manryo Iwayanagi | Yamatsutsuji | Shimotsuke | Aoki Shashanpo | Hagi 311 NS ae | Aus den gegebenen Tabellen ist erkenntlich, dass Arten und Zahl der Pflanzen auf den Fliichen durch das haiufige Brennen der Hara nach und nach vermindert werden. Auf jeder Hara kommt das Susuki oder Kayagrass im Vorrang vor, weil es cin starkes Anpassungsvermégen_ hat und tiberall noch gedeiht, wo der Boden durch Zufall verwiistet und Gewiichse nicht mehr gut zu wachsen vermdégen, es macht eben geringsten Anspriiche an den Boden. Auf selten gebrannter //ara fiillt der Procentsatz an Kaya ab, d andere dice agegen findet man grosse Mengen von Baum- und Halbbaum-Arten, namentlich 212 O. Shishido: Quercus glandulifera, sowie auch mancherlei Grassarten, welche man auf oft gebrannter //ara kaum antreffen wird. //ava-Boden welcher lingere Zeit nicht gebrannt wird, geht in seine urspriinigliche Vegetationsform (den Wald) wieder tiber. Sengen (Nordseite). (Urwald) (Neigung = 35°) (Grassarten der ganzen Nordseite) (Datum 15/4 u. 28/4) lateinischer Name. japanischer Name, Ordnung, Viola silvestris Kit. var, grypoceras A. Gr, Tachitsubosumire Saxifraga contusaefolia S. et Z. Prenanthes acerifolia Maxim. Cardiandra alternifolia S, et Z. Carex brunnea Thunb, Tricyrtis hirta Hook, Ligularia japonica Less. Polygonatum lasianthum Maxim. Aspidium lacerum Sw. Heloniopsis japonica Maxim. Aconitum Fisheri Reich. Ophiopogon japonicus Ker, Ainslizea acerifolia Sch, Bip. Carex Morrowi Boo't, Angerica polymorpha Maxim, Aspidium erythrosorum Fat. Carex duvariana Fr, et Say. Disporum sesile Don. Ifymenophyllum barbatum Bak, Senecio Krameri Fr, et Sav. Astilbe Thunbergii Miq. Daimonjiso Fukuw6so Kusaajisai Nakirisuge THototogisuso Hankaiso Miyamanarukoyuri —. Kumawarabi Shirobana-shojobakama Torikabuto Janohige Momijihaguma Kansuge Shiranesenkiu Benishida Kesuge Hochakuso Koyakokeshinobn Yaburegasa oriashishiGma nach der Haufigkeit in abstergender Skala geord- net, | Ueber die Einwirkung des Hara-Brennens, 313 Gentiana scabra Bge. var, Buergeri Maxim, Rindo Aster japonicus Miq. Yamashirogiku | Anemone hepatica L, Suhamas6 Cacalia delphiniifolia S, et Z. Momijigasa Carex conica Boott. Ilimekansuge Cirsium spicatum Maxim, Yamaazami Cypripedilum japonicum Thunb, Kumagaiso Hosta Ccerulea (Andr) Tratt. Gibdshi Cimicifuga japonica Spr. Plagiogyria enphlebia Mett. Peris cretica L, Obainomotos6 Clematis japonica Thunb, Hanshozuru Calantha reflexa Maxim, Natsuebine | Cypripedilum debile Kchb. f. Koatsumoriso Arisaema japonicum BI. Tennansho | Asarum Blumei Duch, Kanaoi | | Kikenshima Kijinoo Calanthe discolor, Lind}. Ebine Lycopodium serratum Thunb, Togeshiba Cymbidium virens Lindl, Shunran Sengen (Siidseite). (Urwald) (Neigung = 34°) (Grassarten in der ganzen Siidseite) (Datum 17/4 u. 28/4) lateinischer Name, japanischer Name, Ordnung, Ainslizea acerifolia Sch. Bip. Momijihaguma Carex conica Boott. por era pai in | abstetgender | Skala geord- Aspidium aristatum Sw, Hosoba kanawarabi Angelica polymorpha Maxim, Shiranesenkiu net. Saxifraga contusaefolia S, et Z, Carpesium cernuum L, Himekansuge | nach der Sajigankubiso | Daimonjiso | ‘ 314 0. Shishido: Caidiandra alternifolia S. et Z. Kusaajisai Ophiopogon japonicus Ker, Janohige Cryptogramme japonica Prantl. Kanshinobu | Lilium cordifolium Thunb, Ubayuri | Woodwardia radicans Sw, var. orientalis Ltrs, Komochishida . Asarum Blumei Duch, Kanaoi Arisaema japonicum Bl. Tennansho Hosta Sieboldiana (H, K.) Engl. Togibdshi | . Polygonatum lasianthum Sat. Maxim, Miyamanarukoguri | Adianthum monochlamys Sat. Hakoneshida Carex Morrowi Boott. Kansuge Calanthe discolor Lindl. Ebine Cirsium spicatum Maxim, Yamaazami ‘ Cymbidium virens Lindl. Shunran Astilbe Thunbergii Miq. Toriashishoma Tris japonica Thunb. Shaga Senecio Krameri Fr, et Sav. Yaburegasa Aster japonicus Miq. Ni amashirogiku Disporum sessile Don. Tidchakus6 Cimicifuga foetida L. var, simplex Huth. Sarashinashoma dl Anemone hepatica L, Suhamas6 Solidago Virga-aurea L, Awadachiso Heteropappus hispidus Less, var, isochaetus Fr, et Sav. Yamajinogiku Patrinia scabiosaefolia Link. Ominaeshi Chimaphila japonica Miq. Umegasaso Rhizogonium Dozyanum Lacost, Ttachinoshippo Usnea longissima Ach, Saruogase Im Wald “Sengen” gibt es also nur geringe Mengen von Grass aus natiirlichen Grunden und jene Lichtgriser, welche auf jahrlich gebrannter lara vorherrschen, k6nnen hier nicht gefunden werden. Beim Eintritt in einen geschlossenen Wald wird man wahrnehmen, dass fast kein Grass auf dem Waldboden sich findet, wahrend in geringer eeschlossenen Bestiinden das Grass sich mehrt. Der Grund davon ist, darin Ueber die Einwirkung des Mara-Brennens. 315 zu suchen, dass die Sonnenstrahlen in geschlossenem Walde, das ganze Jahr hindurch den Boden wenig erreichen kénnen. Die Griiser, welche in der jungeren Zeit der Bestinde noch zahlreich sind, werden durch den sich verdichtenden Schluss der Bestande von Jahr zu Jahr weniger, da ihnen die zur Existenz néthige Lichtmenge allmahlig zu fehlen beginnt; Aus gleichen Griinden kommen auf der Hara, welche tiber 10 Jahre nicht gebrannt ist und einen grésseren Antheil an Baumen Striiuchern anfweist, verhaltniss- missig wenige Griser vor. 1. Verminderung der Gewdehsarten. Die Tabellen zeigen ferner dass die Menge der Gewichsspecies auf oftmals gebrannter Hava abnimmt, wahrend sie in der selten gebrannten Hara zunimmt, bis auf einer Hara die Baumflora vorherrschend wird. Ge- wachsarten, welche durch hiaiufiges Brennen auf einer Hara an Zahl abnehmen, sind : Disporum sessile Don, TGchakaso Salmia japonica var, buergeri Maxim. Akinotamurasd Pieris hieracioides var, japonica Rge. KGzorina Carex brunnea Thunb. 4 Nakirisuge Tupatrium kirilowii Furez. Sawahiyodori Eupotrium japonicum Thunb, Hiyodoribana Asarum blumei Duch. Kanaoi Polygonatum giganteum var. thunbergii Max, Narukoyuri Clematis heracleifolia var, Stans, Kusabotan Chrysanthemum sinense var, japonica Max. | Ryundgiku Ixeris thunbergii A. Gr, Nigana Artemisia japonica Thunb. Otokoyomogi Kraunhia floribunda Taub. [Fuji] Dianthus superbus L, Kawaranadeshiko Tricyrtis hirta Hook. Hototogisusd Phagopteris totta Mett. Mizoshida Aster japonicus Miq. Yamashirogiku 316 0. Shishido: Carex conica Boott. Potentilla fragarioides L. Carpesium cernuum L, Cardiandra alternifolia S. et Z. Artemisia vulgaris var. indica Max, etc. Himekansuge Kijimushiro Gankubiso Kusaajisai Yomogi ete: 2. Unter dem Brennen nicht letdende Gewdchsarten. Sie sind folgende: Miscanthus sinensis Anders. Potentilla fragarioides var, ternata Max, Serratula atriplitifolia B. et H. Sanguisorba officinalis L. Thalictrum minus var, elatum Lecoy. Lysimachia clethroides Duby. Plectranthus inconspicuus Miq. Pteris aquilina L. Senecio krameri Fr. et Sav. Polygonum cuspidatum 5. et Z. Euphorbia siebo'diana Mor, et Dic. Imperata arundinaceae var, Koenigii, Astilbe thunbergii Miq. Gerbera anandria Sch, Bip. Athyrium nipponicum §, et Z. Atractylodes lyrata S, et Z. Dioscorea tokoro Makino. Arundinella anomala Steud. Cai (er, Susukt Mitsubatsuchiguri Kumatoribokuchi Waremoko Akikaramatsu Okatoranoo Yamahakka Warabi Yaburegasa Ttadori Nats utda: i Chigaya ToriashishOma Sebonyari Inuwarabi Okera Onidokoro Todashiba etc: diese Grassarten sind eben jene Sichtgraser, welche nur auf alljaihrlich gebrannter //ara sowie auch auf anderem beschidigten z. B. verhagelten Boden noch gedeihen. Ueber die Einwirkung des Hava-Brenneus. 317 (B) Grasarten der verschiedenen Standorte. 1. Grasarten auf dem Riicken der Berge. Gerbera anandria Sch, Bip. Senbonyari Sanguisorba officinalis L. Waremoko6 Serratula atriplicifolia B. et H. Kumatoribokuchi Potentilla fragarioides var. ternata Max, Mitsubatsuchiguri Pteris aquilina L, Warabi Lysimachia clethroides Duby. Okatoranoo Imperata arundinacece var, Koenigii. Chigaya Astilbe thunbergii Miq. ToriashishOma Brachypodium silvaticum R. et S. Yamakamojigusa Euphorbia sieboldiana Mor, et Dic. Natsutodai Euphorbia Onoei Fr, et Sav. Takatodai Arundinella anomala Stiud. Todashiba Agrimonia viscidula var, japonica Miq. Kinmizuhiki Phytolacca acinosa var. esculenta Max. Yamagob6 Taraxacum officinale Wigg. var, glaucescens. Tanpopo etc. ete, Diese Arten benétigen zu ihrem Gedeihen eine geringere Feuchtig- keitmenge. Auch sind sie gegen extreme atmosphirische Einwirkungen weniger empfindlich. 2. Grasarten am Mittelhange. Senecio krameri Fr, et Sav. : Vaburegara Polygonatum cuspidatum 5, et Z. Itadori Thalictrum minus L, var. elatum Lecoy. Akikaramatsu Aster indicus L, Yomena Osmunda regalis L, var, japonica Milde, Zemmai Plectranthus graucocalyx Max, var. japonica Max. Hikiokosht Aster scaber Thunb, Shirayamagiku Patrinia scabiosaefolia Tae Patrinia villosa juss, Seseli livanostis var. daucifolia DC. Artemisia japonica Thunb, Cursium spicatum (Maxim.) Miscanthus sinensis Anders. Solidago virga aurea L, 0. Shishido: Ominaeshi Otokoeshi Tbukibéfu Otokoyomogi Yamaazami Susuki Awadachiso Serratula coronata L, Tamaboki Angelica decursiva Miq. Nodake Plectranthus inconspicuus Miq. Yamahakka etc, ete. 3. Grasartenim Thale. Aralia cordata Thunb. Udo Petasites japonicus Miq, Fuki Oenantke stolonifera DC. Seri Acorus gramineus Ait. Sekisho Astragalus sinicum L, Rengeso Tris japonica Thunb. Shaga Equisctum arvense L, Sugina Clematis herachipolia var, Stans S, et Z. Kusabotan Vicia hirsuta Koch, Suzumenoendo Achillea sibirica Ledeb. Nokogiris6O Ranunculus acer var, japonicus Max, Umanoashigata Osmorhiza japonica $, et Z. Yabuninjin Rumex japonicus Meiser. Gishigishi Angelica miqueliana Max. Serimodoki_ Geranium nepalense Sweet. Furoso Carex confertiflora Bott, Shirasuge Carex gibba Wahl, Maskkusa ete. ete. Vorgenannte Griaser sind hauptsachlich auf alljahrlich gebrannter Hara wahrzunehmen. Ueber die Einwirkung des Harva-Brennens. 319 Am Bergkamme gedeihen nur diejenigen Grassarten, welche grosse Menge von Sonnenlicht zu ihrem Wachstum benéthigen; wahrend im Thale jene Arten fortkommen, welche zu ihrem Gedeihen weniger Sonnen- licht dagegen mehr Feuchtigkeit nothig haben. Beim Vergleich mit wenig gebrannter Hara zeigt sich nun die auffallende Thatsache, das auch jene Grasarten, welche sonst nur geringere Sonnenlichtmenge beanspruchen hier ebnfalls am Kamme gefunden werden. Die Eigenschaften resp. Exis- tenzbedingungen der Griser am Mittelhangen haben entsprechend ihrer Lage weniger ausgesprochene Besonderheiten. Il. KAPITEL: Wauchszustinde der Gréser auf gebrannter Hara. Auf selten gebrannter Hara ist die Bodendecke dichter, sic schiitzt den Boden vor direkter Einwirkung der Atmosphiirilien und gibt in den Verwesungsprodukten ihrer Pflanzen gewissermassen den Diingstoff fiir dic spatere Generation wieder. Solche Bodendecken enthalten eine grosse Menge von Feuchtigkeit, u. verhindern weiterhin eine starke Austrocknung des Bodens durch die Sonnenstrahlen, unter solchen gunstigen Umstanden leben dann auch Gewachse, welche zu ihrem Gedeihen eine gwewisse wenn auch miassige Menge von Nahrstoffen und Feuchtigkeit unumgiinglich nothig haben. Auf nackter Hara kénnen nur bestimmte Pflanzen leben, denen neben starkem Sonnenlicht, hohe Wiarme und ein sehr geringer Procentsatz an Nihrstoffen und Feuchtigkeit néthig ist. Wenn man z. B, die Entwickelungsphasen von Miscanthus sinensis, Pteris aquilina, Potentilla fragarioides, Euphorbia sieboldiana, und Sanguisorba officinalis autmerksam beobachtet, so mégen sie oft zwar zahlreich vorhanden, aber arm an Arten sein, sowie von sehr kleinem oder zwerghaftem Wuchse. Ich unter- suchte das Lingenwachstum des Artwuchses auf verschiedenen Hara’s. Solche Beobachtungen miissen natiirlich in der Zeit gemacht werden, wo die Grass-Pflanzen ihre Lebensthitigkeit eben vollendet haben. Die Entwickelungszustinde der betr. Gewiichse sind im Einzelnen je nach- den Standortsverhiltnissen und Jahreszeiten ganz verschieden. So z. B. die Lange-Entwicklung von Miscanthus sinensis. Dieses hat im Spitherbst 320 (0. Shishido: des vorigen Jahres seine Lebensthatigkeit vollendet und steht jetzt (am Ende April) absterbend auf der Hara. Ich dehnte meine Messungen sowohl auf die 7ékrlich gebrannte Hara “ Kiridéschi,” als auch in auf “Qmiyama” (woselbst seit 3 Fahre nicht gebrannt wurde) und auf bd ‘Nanamagari’’ woselbst seit 1o Jahren kein Brenner. stattfand, aus Kiridéschi (Nordseite) Linge (das Mittel von 100) Kamm 1.6033 m. Mittelhang 1.9433 m Thal 2.0900 m. Mittel 1.8789 m. Omiyama (Nordseite) Linge (das Mittel von 100) Kamm 1.9013 m. Mittelhang 2.3200 m. Thal 3.0833 m Mittel 2.4349 m. Nanamagari (Nordseite) Lange (das Mittel von 100) Kamm 2:21.35 m- Mittlhang 2-5OLS 1. Thal 3-4022 m. Mittel 2.7057 m. Fir andere Grasarten konnte ich kein passendes Untersuchungs- material finden; an der kleinen Untersuchung tiber Miscanthus sinensis jedoch lisst sich der ungiinstige Einfluss des jéhrlichen Brennens schon deutlich erkennen. Richtiges, dickstengeliges Kayagras kann man nur auf selten gebrannter Hara antreffen. Sn ae ee Ueber die Einwirkung des Zara-Brennens. 321 IV. KAPITEL: Verdanderung der Bodenbeschaffenheit durch das Brennen der Hara. (A) Verdnderung der phystkalischen Eigenschaften des Bodens. Die Pflanzen bilden sowohl als lebende Individuen, wie auch durch ihre abgestorbenen KG6rpertheile unter den gew6hnlichen Verhialtnissen eine Decke, fiir jede Bodenart. In dieser Decke finden dann andere Pflanzen wiederum ihr Gedeihen. Es tiben zweifellos verschiedene Decken auf den Pflanzenwuchs und den Boden einen giinstigen oder ungiinstigen Einfluss je nach dem aus. Auf viel gebrannter H/arafliche z. B. in “ Kiyosumi,” wo man durch das Feuer die Pflanzen-Decke des Bodens oft vernichtete, inderten sich alle Verhiltnisse besonders die physikalischen Eigenschaften des Bodens in ungiinstiger Weise weil er nunmehr schutzlos den atmosphirischen Einwirkungen direkt wiederholt ausgesetzt wurde. Die flach- und mittelseit griindigen Bodenarten finden sich in der Regel, entweder auf den Riickenplateaus oder an den oberen Gehingen der Gebirge, namentlich wenn die letzteren waldlos sind; der in den unteren Regionen und am Fusse der Gebirge lagernde Boden dagegen ist mittel- ’ griindig oder sogar tiefgriindig; inbesondere bei steilen Gebirgsgehangen wird dies deutlicher erkennbar. Das kann auch auf der Hara von Kiyosumi nachgewiesen werden, besonders auf der haufiger gebrannten //ara. Ich habe schon angedeutet, dass die Gebirge in Kiyosumi sehr steil sind, meist tiber 35° Neigung ; wird nun die Pflanzen- Decke auf der Hara durch hiufiges Brennen wiederholt fast gianzlich vernichtet, so gelangt der feine gute verwitterte humose Boden immer mehr durch die Regengiisse y yi in grosser Menge thalwirts, so dass die einen Riicken bekleidende Hara (d. h. deren Boden) sich als ausserordentlich flachgriindig u. nahrungsarm nachweisen lisst; nicht selten tritt sogar bald das Grundgestein zu Tage. Im Thale dagegen ist durchgiingig der Boden tief- ja oft sogar sehr tief- griindig zu finden. Hara-boden muss also umso seichtgriindiger werden, je Ofter die Hara gebrannt wird, Auf selten gebranntem Hara-boden wird durch die lebenden 322 ©. Shishido: und abgestorbenen Pflanzentheile der Boden gehalten, was in grésserer Tiefgriindigkeit zum Ausdruck kommt. (B) bendigkeit. Wie erwahnt sind im Allgemeinen die Verwitterungsb6den der Tertiarperiodengesteine zu den fruchtbaren zu rechnen. Sobald aber, wie auf der Kiyosumi //ara, der Bodeniiberzug 6fters abgebrannt wird, tritt auffallend rasch Verschlechterung ein; alljahrlich gebrannte Hara, zeigt sehr lockeren oder oberflachlich grob gekrimelten Boden, welcher gegen Pflanzen- Wuchs sich ungiinstig verhalt infolge seiner grossen Austrock- nung. Weiterhin hangt die Empfanglichkeit eines Bodens fiir die Erwarmung durch die Sonnenstrahlen zum gréssten Theile von der Beschaffenheit seiner Oberflache ab; in dieser Bezichung gilt, dass, je grobkérniger das Gemenge eines Bodens ist, desto schneller und starker er sich erhitzt durch die Insolation, aber dass er auch um so schneller die hierdurch erhaltene Warmesumme wieder an die Atmosphire abgibt, sobald die Sonne seine Oberflache nicht mehr bescheint; je fezxkriimeliger oder dichter dagegen ein Boden-Gemenge sich zusammenstezt, umso langsamer erwarmt es sich, aber um so langer halt auch der Boden die einmal aufgenommene Warmemenge fest. Der erste Fall ist zweifellos dem Pflanzengedeithen sehr schadlich, wahrend der /etztere als giinstig bezeichnet werden muss. Da die selten gebrannte Hara den feingekriimelten und milden bindigen Boden enthialt, so gedeihen daher auch die Pflanzen auf dieser Hara besser, wahrend die jahrlich gebrannte //ara mit ihrer schlechten Boden- struktur, nur schlechte Gras-Arten, mit extremen Anpassungsvermégen cin Fortkommen gestattet. Wasser ist ein wichtiges Lebensmittel fir alle Pflanzen. Ohne Wasser, keine Pflanzen! Es ist also der Boden, worin die Pflanze wurzelt, nicht . blos ihr Schiitzer gegen Winde, ihr Erwarmer in der Kalte, sondern auch ihr Wasserspender, wenn die Atmosphire ihr kein Wasser darreicht; dieses wichtige Amt vollbringt der Boden einerseits durch sein Wasseransaugungs- vermoégen und anderseits durch seine Wasserhaltungskraft ; das erste ist ul oo ie Og CE ae bi, Ueber die Einwirkung des ZZara-Brennens. 3 to Ww eine Thiatigkeit der Capillaritat, welche sich umso giinstiger bemerklich macht, je feinkrumiger der Boden ist, im zweiten Sinne wirkt am besten ein an Humus- und Thonsubstanzen reicher Boden. Den besten Standort fiir die meisten Gewachse bieten diejenigen Bodenarten, welche unter den gewodhnlichen Verhiltnissen bei starker Wasser- Aufsaugung das in ihnen angesammelte Wasser missig festhalten und missig verdunsten. Auf der Kiyosumi Hara aber, dort wo das Gras jahrlich gebrannt wird, kann der Boden diese wichtige Aufgabe nicht leisten, weil die Decke ja beinahe ganzlich vernichtet wird und der Boden direkt den atmosphirischen Einwirkungen preisgegeben wird. In der Oberflachenstruktur des Bodens sind grobe Kriimmel keineswegs dem Pflanzengedeihen zusagend. Je hiaufiger die Hara gebrannt wird, desto mehr trocknet der Boden aus und desto gréber wird seine Struktur durch Auswaschung werden; es herrschen Extreme von Trockniss und Feuchtig- keit in solchem Boden. Auf anderer nur selten zum Brande gelangter Hara ist der Boden dem Pflanzenwuchse zusagender, weil feinkorniger und mit grossem Wasser- Aufnahme u. Erhaltungs- Vermégen ausgestattet. (B) Verminderung der organischen Substanzen tm Boden. Die organischen Substanzen im Boden bilden fiir die Pflanzen niitz- liche ja unenthehrliche Stoffe; man nennt daher einen Boden, weicher an organischen Substanzen reich ist, einen produktiven Boden. Die organischen Substanzen im Boden entstehen durch langsame Oxydation abgestorbener Pflanzentheile der Bodendecke unter dem Einflusse der Bodenfeuchtigkeit und werden “ Hummus” genannt. Auf solcher Hara, wo das Gras alljahrlich gebrannt wird, vernichtet das Feuer fast die ganze Bodendecke mit den Humusstoffen, welche die Pflanze so néthig zum Wachsthum fat. Bei meinen Untersuchungen in Kiyosumi stiess ich demgemiiss stets auf nachtheilige Wirkungen des wiederholten HYava-Brennens. ‘UsTye7Z apussjoy qes1o Uspeg UdSIDAIP Usp UL UdzZURYsqnuS JOYysIUeSIO UNSUD;Y JOp SunyonsioyUQ VI “epiNM yuURIGIS [eUyo v4VET dUID UNDA “IeqsIOMYILU Ie]Y ‘Uopoq-v4vz_F sop YyesyssunyeyJosseA\ Jop SunJopulu4sA sIp ys! 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Sane be ‘ == gee Feinerde, | ausgetrockneter | bindungswasser _bindungswasser 3 st. aah en ee in Feinerde, im Originalboden, (B—A) Feinerde. & Se ahs SS Kiridéschi (Nordseite) 0.3790 gr, 14.4513 % 12.6333 % 12.8331 % a (Stidseite) 0.3792 ,, 14.1889 ,, 12.6400 ,, 12.2583 , Kajisaka (Nordseite) 0.3994 ,, 15.2956 ,, 13.3133, 12.3338 ss (Stidseite) 0.3734 5; TA S220. 3 12.4467 ,, 12.1343 Omiyama (Nordseite) 0.3976 ,, 18.6567 ,, 16.5900 ,, 15.9630 ., 53 (Stidseite) 0.3956 ,, 14.8276 ,, 13.1866 ,, 12.8913 326 0. Shishido: Musado (Siidseite) 0.3966 ,, NS AUS Olas 13'2200 15, 12/Q2 0500s: Omiyama s 0.3893», rise Gee 12.9766 ,, 127220 —. Suzuriischi + 337780555 1423122) 55 12.6000 ,, 12.2509 ,, Sannodai _ (Nordseite) 0.4199 ,, W5tOS mess 13.8966 ,, 13.4984 ,, Ai (Stidseite) 0.3609. ,, 13.6075 3; 12.0300 ,, 11.7641 ,, Suzuriichi (Nordseite) , 0401! 5, TS 5550; 138 700Nss 13.2304. 5, Nanamagari 0.4180 ,, 16.3020 ,, Tea.0323me" U3 7O5At ts Musado 5 O:M4ARR EE. 1y/A0 LOO ss 14.8433 > 14.3817 Fe Sengen = O:5L1Om., 19.5411 ,, 19,0333 is 16.8426 ,, x (Stidseite) 0.5069 5, 19.8631 ,, 16.8966 ,, 16.6040 ,, Aus der Tabelle kann entnommen werden, dass in jahrlich gebranntem Hara-boden eine starke Verminderung der organischen Substanzen eintritt, wahrend in selten gebranntem Hara-boden diese zunimmt. Vill. Abschnitt. Die Einwirkung des Hara-brennens in dkonomischer Bestehung. In den meisten Gegenden herrscht der Glaube, dass durch das Feuer die Wachsthumskraft der Hara _ gesteigert werde. Diese Annahme scheint nur auf ebenem Felde als richtig, wahrend sie an_ steilem Abhange nicht zutreffen kann, da im letzteren Falle die Pflanzen- Asche nicht langer am Hange bleibt, sondern durch die Regengisse thalwarts gefiihrt wird. Man hat eben nur nach bestimmten Artem gesehen, welche sich unter Umstinden auf alljahrlich gebrannter Hara in der Mehrzahl finden, aber nicht nach der allgemeinen Beschaffenheit der Bodendeckenvegetation und ihren Wuchszustinden. Miscanthus sinensis, welches den Leuten als Dach-Deckmittel unentbehrlich scheint, kann auf gebrannter Hlara zwar in Masse vorkommen, doch wenn man genauer zusicht, so findet man, dass der einzelne Stamm kiurzer und diinner ist, als auf anderem Platze, der selten gebrannt wird. Diese Thatsache beweist, dass das Brennen seinen Zweck nur mangelhaft erfiillt, es kann solch zwerghafte Ueber die Einwirkung des Hara-Brennens. Miscanthus dem Bediirfnisse der Leute die richtigen und langen Halme gew Auch die Griiser von alljahrlich gebr auf die Dauer nicht gerecht werden: innt man nur auf ungebranntcr Hara. annter Hara sind vorzugsweise “saure”’ Graser und fiir das Vieh anim gentessbar.. Betrachtet man also die Sache Brennen der Hara nichtsweiter als ej unbefangen von allen Seiten, so ist das ne schlechte, durch Nichts begriindete Gewohnheit und « unwirthschaftlich YS 2 nennen, weil der Boden, dem sein natiirlicher Humus genommen wird, successive zur Inproduktivitit herabsinkt. In Kiyosumi hat man sich in Erkenntniss dieses Umstandes zur Aufforstung in planmissiger Weise entschlossen, da dadurch ein weit grosseren Gewinn erzielt werden wird, als bei der bisherigen Hara- wirthschaft, welche fiir das Land nicht die mindeste wirthschaftliche Bedeutung, hat sondern lediglich als eine “ schlechte Gewohnheit’ werden muss. : bezeichnet XT. Abschnitt. Die Einwirkung des Brennens der Hara im landwirthschaftlichen Sinne. Das Hara-Brennen in Japan entstand hauptsachlich aus den Bediirfnissen der Landwirthschaft, man wollte dadurch eine grosse Menge Griindiinger bekommen, aber das Brennen der Hara hatte wie iiberall so Kiyosumi Hara nur das Gedeihen “ schlechter” welche fiir den Boden als Diingmittel k auch auf der Grasarten, zur Folge aum wirksam zu nennen sind. Auf jener Hara dagegen, wo selbst das Gras sehr selten gebrannt wird, finden sich gréssere Mengen von guten Species unter anderm z. B. Legminosen arten in grosser Zahl. Diese Letzteren enthalten gro sse Mengen von Stickstoff, welche sie aus der Luft absorbieren, wodurch sie als Diinemittel sehr wirksam sind. In der Asche, welche durch das Brennen der Bodeniiberzuges erzielt wird, finden sich zwar fiir die Pflanzen einige gute Nahrstoffe, z. B. das | p . } ; Kali; aber an steilen Abhingen, wie auf der Kiyosumi Hara wird es 328 ©. Shishido: zusammen mit den feinen produktiven Bodentheilen durch die Regengiisse thalwarts gewaschen und dadurch erst recht ein nahrungsarmer Boden geschaffen. Als Futtermittel sind jene schlechten sauren Grasarten, welche sich auf (durch das Feuer) oft verwiisteter Yara befinden, fiir das Vieh kaum verwendbar, wahrend die auf guter, bezw. selten gebrannter Hara wachsenden Arten als ein sehr gutes Futtermittel bezeichnet werden miissen. Durch das Brennen der Hara werden also gute nahrungsreiche Grasarten vernichtet und der Boden landwirthschaftlich werthlos gemacht, die Hara erfillt also gerade ihren eigentlichen Zweck umso weniger, je ofter sie gebrannt wird. xX. Abschnitt. Die Linwirkung des brennens der Hara in forstlicher Hinsicht. I. KAPITEL: Gefahrlichkett des Brennens der Hara. Die Forstwirthschaft bedarf im Allgemeinen grosser Vorrath-Kapitalien und langere Zeitraume zu ihrer Produktion, um gute Ertrage nachhaltig in ihren Waldungen abzuwerfen. Unter verschiedenen Gefahren wie Insekten, Sturm etc, welche den Wald bedrohen, ist auch das Feuer zu erwahnen. In Japan sind Waldfeuer sehr hiufig, was hauptsichlich auf die achtlose Behandlung des Feuers zuriick zu fiihren ist, namentlich ist es das Brennen der Hara, welchem grosser Schaden im angrenzenden Wald alljahrlich zur Last zu schreiben ist. Die Hara wird hauptsiachlich im Frihjahre (von Februar bis Ende Marz) gebrannt; die an solche Hara angrenzenden Waldtheile stehen in grosser Gefahr. Es gibt zwar gesetzliche Vorschriften ttiber das Brennen der Hara, wonach die Hara ohne zuvor erholte Erlaubniss tberhaupt nicht gebrannt werden soll, auch muss dem benachbarten Grundstiick ein gewisser Schutz gegeben werden durch Zichen eines sogenannten Schutzgrabens. Da diese Vorschriften nie beachtet werden, tritt das Feuer tiber die Schutzzone in den benachbarten Wald nur zu oft iiber. Ueber die Einwirkung des Hara-Brennens. 329 Die Hara in Kiyosumi grenzt an ihrer ndrdlichen Seite an das Schulwald-Areal an, welches in letztes Zeit aufgeforstet wurde; es bedarf deshalb einer besonderen Aufsicht, wenn man das Brennen gestatten will. Die Vernichtung von Wald ist eine constante Begleiterscheinung des Brennens der Hara und auch aus diesem Grunde méchte diese Boden- nutzungsmethode zu verwerfen sein. Il. KAPITEL: Verunichtung der Bodendecke. Wie ich schon bemerkt habe, bildet die Pflanzen- Vegetation durch die Humusbildung eine Decke, welche der Erhaltung jeder Verwitterungs- schicht giinstig ist. An abhingigen Lagen bildet sie einen das Fortschlim- men der Erdtheile verhindernden Widerstand und an sonnigen Stellen schiitzt sie den Boden gegen die Sonnengluth und bewahrt so den von ihr bedekten Boden vor Ausdo6rrung, im Sommer, wie vor den Einwirkungen des Frostes im Winters. Sie lasst starke Regenmengen nicht direkt zum Boden dringen, sondern gewihrleistet deren allmahlige und mehr gleichmiassige Aufsaugung in den Boden. Das zu rasche Abfliessen des Wassers mit seinen schadlichen Folgen wird hauptsichlich verhindert. Durch das alljahrliche Brennen von Hara-flachen wird nun die Humus- und Pflanzendecke ginzlich vernichtet, sie kann also auch ihr wichtiges Amt nicht mehr vollbringen. Ill. KAPITEL: Verminderung der Nahrungsstoffe im Boden. Man unterscheidet zwei Arten von Nahrungsstoffen im Boden namlich die organischen und anorganischen oder mineralischen Stoffe. Die org- anischen Substanzen im Boden enstehen durch den Verwesungsprocess der abgestorbenen Organismen (d. h. Pflanzen und Thiere). Auf Harv und Waldboden sind sie meist von abgestorbenen Pflanzentheilen gebildet. Der Boden enthilt cine um so gréssere Menge von Humussubstanzen, je grdsser die Zahl der dort lIebenden und absterbenden Pflanzen ist; auf Hara aber, welche alljahrlich gebrannt wird, gibt es nur eine geringe Menge abgestorbener Pflanzen; auf der schutzlosen Hara bringen 330 0. Shishido: die Regengiisse etc. unaufhérlich Humussubstanzen, feinen Sand und erdige Theile von den Berggehangen herab in die Thaler zur Ablagerung oder » in die Bache zum weiteren Transport; wenn man also an die Aufforstung oft gebrannter Hara geht, so fehlen dem Boden jene Nahrungsstoffe, welche zum Gedeihen der jungen Bestande so notwendig sind; man kann daher auf solchen schlechten Béden auch nur jene Arten von Baumen pflanzen, . welche die geringsten Anspriiche an die Nahrkraft des Bodens stellen, auf die besseren Baumarten muss man also von vorn weg verzichten, was einem erheblichen wirtschaftlichen Nachtheil gleichkommt. Auf die traurigen Folgen der Geschiebefithrung der Bache und Fliisse infolge der Hlara-bedeckung der Gebirge sie hier nur kurz hingewiesen. IV. KAPITEL: TJvockenheit des Bodens. Wo dem Boden die néthige Menge von Wasser fehlt, da bleiben die Nahrungssubstanzen in ungeléster Form, was wiederum Nahrungsnoth fur die Pflanzen bedeutet; auch kénnen die Pflanzen ihre zu ihrem Gedeihen nothige Transpirationsgrésse bei Wassermangel nicht aufrecht ertalten u. sterben ab. Ist nun der Boden mit Pflanzen und Humusdecke geschiitzt, so kann er eine bedeutende Wassermenge aufnehmen u. festhalten, welche langsam aber sicher u. wirksam den Pflanzen zu Gute kommt, wahrend auf nackten Hara’s die meteorischen Niederschlage oberflachlich rasch abfliessen und der Boden der Trockenheit ausliefern. Trockenheit des Bodens aber ist fiir die Forstwirtschaft oft ein sehr grosses Hinderniss; gute Bestiinde kénnen nur auf jenen Béden gedeihen, wo ihnen eine gewisse Menge Feuchtigkeit nicht fehlt. Nachdem die Hara in Kiyosumi, alljahrlich gebrannt wird, hat sie auch nur eine schwache Pflanzen- & Humusdecke. Aus diesem Grunde kommt dann der fast nackte Boden durch Einwirkung der Atmosphire, namentlich durch die starke Besonnung mehr u. mehr herab. Wollte man solchen Boden sofort aufforsten, so wiirden die Pflanzen die néthige Wasser- und Nahrungs- menge im Boden kaum finden und zu Grunde gehen. Dagegen hat man - < stets leichteres Spiel mit jener Hara, welche seltener gebrannt ist, sie setzt der Aufforstung viel weniger Schwierigkeiten entgegen. Veber die Einwirkung des MWara-Srennens. we ir — Schiuss. Aus den bisher mitgetheilten Beobachtungen mag ersehen werden, dass die Kiyosumi Hara, da sie in der subtropischen Zone mit deren grosser Menge an Luftfeuchtigkcit bei seltenem Frost und Schnee liegt, in Bezug auf das Pflanzengedeihen zu den giinstigen Platzen gerechnet werden muss. Trotzdem wird zur Zeit von den grossen Flachen der Kiyosumi Hara in ihrer “ Hara’’—Form, nur geringer Nutzen gezogen, sie sind de facto Nichts weiter als unproduktive Flachen. Oftmaliges Brennen der Hara verursacht folgende Nachtheile : 1. Die Pflanzen, Bodendecke ist durch das hiufige Brennen der Hara fast ganz zum Verschwinden gebracht. lo der Boden wird direkt den Atmosphirilien greisgegeben. 3. die Regenwasser fliessen oberflichlich ab und verursachen keine Zunahme der Bodenfeuchtigkeit. 4. der Boden geht in grobe Kriimmelstruktur iiber. 5. die produktive Bodenkrumme wird durch die Regengiisse thalwirts abgefiihrt. 6. Die Bodenverwitterunsgschicht wird dadurch seichter und seichter. 7. Es tritt im Laufe der Zeit Humus-Verarmung im Boden ein. 8. die Lichtgriiser kommen auf viel gebrannter Hare in Uberzahl vor. g. die Grasspecies werden durch das Brennen in ihrer Zahl vermindert. 10. die Gewiichse auf oftmal gebrannter Hara neigen zur Degenerirung. 11. Baumarten verschwinden sobald eine Hara ofter gebrannt wird. 12. wenn eine //ara lingere Zeit nicht gebrannt wird, so geht sie wieder in Wald iiber, zuerst erscheint “ Quercus glandulifera” in hiesiger Zone wieder. 13. das Hauptprodukt der Hara, (d. h. Kaya) Gras, wird durch oftmaliges Brennen weder in richtiger Bonitit noch Menge erzeugt. 14. die Asche, welche durch das Brennen der Griser entsteht, kann auf dem geneigten Hara-boden nicht liegen bleiben, sic hat also als Diinger auch keinen Werth. to Q. Shishido: ioe) oP) 15. das Hara-brennen ist in der Nahe von Waldungen am gefahrlichsten. 16. die Wachsthumsenergie der Graser sinkt durch oftes Brennen herab. Die Hoffnung, durch Brennen Boden in seiner Produktivitat zu heben, ist im Gebirge wenigstens (wie in Kiyosumi Hara) eine irrige. Die japanische Hara, welche die Haupterscheinungsform der- Bodenbenutzung im Berglande ist, verandert den Boden in nachtheiliger Weise, fuhrt Verarmung des ungeschiitzten Bodens durch Auswaschung und Verhagerung iiberall herbei, ja nicht selten tritt das nakte Grundgestein auf japanischer Hara zu Tage. Ich schliesse meine Beobachtungen damit, dass ich behaupte, dass das Flara-brennen vom Standpunkt der Landwirthschaft wie, der Forstwirth- schaft betrachtet, einen entschieden nachtheiligen Einfluss auf die Hara selbst ausiibt, ja dass es geradezu den Hara-boden der Verédung preisgibt. Es waren nach meiner Ansicht die meisten Hara in Japan, weil sie als Hara nicht nur okxe wirthschaftliche Bedeutung sind, sondern eeradezu landverwiistend wirken, a@/sbald aufzuforsten. Wo dies nicht moglich ist, kbnnen gute Graser nur nach Authdren jedes Brennens erwartet werden. + tee at esi > INHALT. Einleitung I Abschnitt Die Geschichte der ‘* Kiyosumi Hara.” IJ. Abschnitt Zustand der Azvyosumi Hara. I. Kapitel: Bodenzustinde. (A) Lage und Fliche. ... (B) Boden (Grund gesteine). II. Kapitel: Das Klima. Ill. Abschnitt Cultur der Gegend bei Kiyosumi. IV. Abschnitt Die Hara und ihre Besitzverhiltnisse. ... I. Kapitel: Besitzarten. II. Kapitel: Gewinnung des Haraprodukte. V. Abschnitt Das Brennen der Hara. ... I. Kapitel: Zweck des Brennens. (A) Entstehung aus alter Sitte. ... (B) Landwirthschaftliche Beniitzung der Hara. (C) Werth des Harabrennens zur Gewinnung von Futter- grasern. ... II. Kapitel: Verfahren des Harabrennens. III. Kapitel: Zeit des Harabrennens. IV. Kapitel: Die Methode des Harabrennens. VI. Abschnitt Einfluss auf physikalische Eigenschaften des Boders. I. Kapitel: (a) Tiefe, (b) Bindigkeit, (c) Feuchtigkeit. II. Kapitel: Aussere Zustinde der Hara. III. Kapitel: Das Verhiiltniss zwischen Boden und Klima (Verwitterungsprocess.) 334 Inhalt. VII. Abschnitt Vergleich der in verschiedenem Brenn-Turnus gebrannten Hara- PIAtzOe jon ca ee OL ee ae As FL 2 er I. Kapitel: Untersuchungen im Einzelnen, Probeflichen. .. 280. Il. Kapitel: Folgen des Brennens der Hara... 2), (A) Verdnderung der Gewichsarten. 30 4 ae 1. Verminderung der Gewachsarten. .. eee 2. Unter dem Brennen nicht leidende Gewiichs- Arten. soe a IO OEL ASE anit (Bb) Grassarten der verschiedenen Standorte. 1.) “... “G1 307. 1. ‘Grasarten aufiden Rucken der Berge. fy 2 een 2. Grasarten'am Mittelhange".. |‘ UR cet Ri ego 3. Grasarten'im Thale.:. =... <2. 22) °)aei gee If. Kapitel: Wuchszustande der Griaser auf gebrannter Hara. ...0 i. ... On | ice IV. Kapital: Veranderung der Bodenbeschaffenheit durch das Brennen der Hara. vie USD eae ae (A) Verianderung der physikalischen Eigenschaften des Bodéns.. 4.2 vou nee) bee tierce = Guee | oe (Bb) Verminderung der organischen Substanzen im Boden. Vill. Abschnitt Die Einwirkung des Harabrennens in 6konomischer Beziehung. — .. 326. IX. Abschnitt Die Einwirkung des Brennens der Hara in landwirthschaftlichem SInNe. «20. 4s. MG eae)! ee ao eS X: ‘- Abschnitt Die Einwirkung des Brennens der Hara in forstliicher Hinsicht. ... 328. I. Kapitel: Gefahrlichkeit des Brennens der Hara. vad i500 980: II. Kapitel:.-Vernichtung ‘der Bodendeeke= \2:.c.7 1a) Same goa Ill. Kapitel: Verminderung der Nahrungsstoffe in Boden. ... 329. IV. Kapitel: Trockenheit:des*Bodens-)\... :i02igke eee Schiuss. — —40>- o—— Die zukunftige Bewirtschaftungsform des japanischen Waldes ! VON Dr. Hefele. Nel. bayr. Forstmeister u. Professor in Toko. oO 2 Die Umwilzung des Jahres 1868 ist auch von grossem Einflusse auf die japanischen Waldungen gewesen. Wahrend friiher wohl die meisten der Feudalherren die ihnen unterstellten Kronforste und Staatswaldungen wenigstens in einigermassen gutem Zustande erhielten, wenn auch cine besondere Waldwirtschaft sich nicht entfaltete, so ist doch in den letzten Jahrzehnten manches Stick trefflichen Staatswaldes der Begehrlichkeit insbesonders biuerlicher Kreise und den schwankenden, wechselnden Auschauungen iiber die Lésung der volkswirtschaftlichen Fragen nach der Restauration in Japan zum Opfer gefallen. Ausserdem besass die neue Centralgewalt vielfach weder die Macht noch die Organe, ihre Autoritat geniigend geltend zu machen, um den durch Sikularisirung von Kloster- giitern, Einverleibung von Gemeindsforsten etc. vermehrten Staatswaldbesitz vor Forstfrevel zu schiitzen. Auch der Privatwaldbesitz unterlag manigfachen Veranderungen. Armuth und Verschuldung vieler Grund-Besitzer, welche bei der Umegestaltung der allgemeinen Verhiiltnisse, sich der Neuzcit nicht anzupassen vermochten, fiihrte hiiufig zum Verkauf, zur Zerstiickelung und Vernichtung von Wildern. Namentlich haben aber Theilungen von Waldgrund unter die ehedem nur zu Nutzungen Berechtigten am vorher gemeinsamen. Walde (Gemeindewalde) zur vorauszuschenden — sicheren Vernichtung, Devastirung oder wenigstens Verschlechterung der Theilstiicke gefiihrt. Auch will mich bediinken, dass die hier auf dem Papier so geordnet und genau festgestellte Oberaufsicht des Staates tiber die Gemeindewaldungen in praxi nicht geniigend strenge zur Anwendung kommt. Der factische Zustand der meisten biiuerlichen Gemeindewaldungen spricht hierftir. 336 Dr. Hefele: Habsucht und mangelndes Verstandniss veranlassen ferner gar manche kleine Privatwaldbesitzer zu einer Nutzungsweise, welche in vielen Fallen direct der Zerstérung gleichkommt. Die Eigenthiimer gvdsserer Privat- waldungen sind bis zu einem gewissen Grade indessen bereits zum tieferen Verstindnisse des Werthes und der Bedeutung des Waldes gelangt und wirthschaften im wohlverstandenen eigenen Interesse etwas mehr conservativ. Am besten wurde der Wald wohl da conservirt, wo die fehlende Aufschliessung der Gegend und die geringe Bevélkerungsziffer ihn von selbst bis heute geschiitzt haben und es gilt dies nicht blos fiir den Privat-, sondern auch fiir den Staatswald. Es mdéchte fraglich erscheinen ob nicht finanzielle Schwierigkeiten des Staates den dort vorhandenen reichen Vorraithen ein schnelles Todesurtheil gesprochen hatten, ware ihre Lage zum Verkehr nur ein bischen giinstiger gewesen. Darin beruht auch heute noch die Gefahr, dass das Bestreben hohe Ertrage wm zeden Preis aus einem Walde herauszuholen, die heiligste Pflicht gegen das nationale Wohl vergessen lasst, nemlich die Sorge fiir die ungeschmiilerte Uberlieferung eines werthvollen Waldbestandes an die Nachwelt durch entsprechende Form der Nutzung & Verjiingung. Liegt ja doch der Werth der Walder nicht blos in ihrem Rentenertrag; die Uberschwemmungen und Wasserkatastrophen, welche der Vernichtung des Bergwaldes z. B. allenthalben zu folgen pflegen, reden eine zu deutliche Sprache, als dass sie misszuverstehen wire. Wohl sind Gesetze zum Schutze solcher Forste erlassen, aber ein Blick in die reale Wirklichkeit zeigt zur Geniige, dass das Wollen dem Ké6nnen noch nicht die Wagschale halt. Der Staatswald leidet abgesehen von einigen grésseren Complexen auch nicht wenig durch die Zersplitterung seines Areales; es wird eine stete Sorge der Regierung sein miissen, cine Verbindung der zerstreuten Theile auf dem Wege des Ankaufes oder Austausches herzustellen. Wenn man sich mit Recht zur Abstossung ciniger sehr abseits liegender Kleinflachen entschliesst, so sollte doch Verschleuderung unter allen Umstinden vermicden werden. Eine intensivere Pflege muss zweifellos durch den néthigen gesetz- miissigen Druck der Gemeindewald im getheilten und ungetheilten Besitze Die cukiinftige Bewirtschaftungsform des japanischen Waldes! 337 erfahren. Auf welche abschiissiger Ebene man sich hierin momentan befindet, zeigt jede Excursion durch das Land. Das Beispiel, welches der Staat den Privaten durch seine cigenc Wirtschaft zu geben berufen ist, setzt aber voraus, dass er sich selbst iiber seine einzuschlagende Richtung klar sei und mit allen Mitteln auch Ernst machen will, sowohl durch die néthige durchgreifende Reformirung seiner Verwaltung als auch durch consequente Befolgung gewisser ]W7rtschaft- grundsitze. Die wichtigste Frage ist hiehei zweifellos jene nach der zweckmissigsten Art der Nutzung und damit zusammenhingend die Untersuchung der den Verhiltnissen am besten gerecht werdenden Ver- jingungs-und Bestandsform. in Ubung sind in Japan in der Hauptsache zur Zeit zwei Hauptnutzungsarten, einmal Kak/schlag auf grésserer Flache, und dann eine Art von auszugsweiser Nutzung, welche /da/schlich den Namen eines Plenterbetriebs erhalten hat. Dem Kahlschlag auf grosser zusammenhingender Fliche musste friiher da, wo durch einen Wass2rlauf die Waldungen giinstige Abfuhrbedingungen gewisse Berechtigung zugestanden D fir ihre Schlagergebnisse hatten, eine werden, immerhin sind die aus ihm hervorgehenden Nachtheile vom waldbaulichen Standpunkte so bedeutend, dass man von ihm iiberall, wo es nur irgend méglich ist, abzukommen trachten wird. Nicht blos sind alle Gefahren wie Frost, Dirre, Unkraut (Bambus) im erhdéhten Masse bei dieser Verjiingungsform gegeben, sondern die nachgezogenen Bestiinde nahezu gleichen Alters auf Flichen von grosser Ausdehnung bilden auch einmal spiter fiir den Wirtschafter sehr schwierig zu realisirende Hiebsob- jekte; umso schwieriger je weniger etwa die Aufschliessung der jetzigen grossen Verjiingungsschlige solcher Plaitze bis zur seinerzeitigen Haubarkeit derselben fortgeschritten sein sollte. Man darf nicht vergessen, dass es nicht Aufgabe ciner Wirtschaft sein kann, nur Wald schlechthin nachzuzichen, sondern die Gegenwart hat die heiligste Verpflichtung, dem Stande des Wissens entsprechend, auch technisch vollkommene Bestandsbilder der Nachwelt zu tiberliefern. Die Gewissensberuhigung, welche vielfach darin gesucht wird, dass man die grossen Schlige mit Nadelholz in irgend einem Verbande durch Pflanzung in Bestockung bringt, ist in vielen Fallen ein grosses Armuths- 8 Dr. Hefele: (Op) 2 eo) zeugniss fehlender’ Uberlegung und Klarhcit itber die Grundziige einer geordneten Wirtschaft. Ziellose Aufforstung gibt wohl Wald, ob aber den nach Betrachtung aller Verhialtnisse rzchtzgex Wald, darnach fragt man, wie mir auf Grund meiner cigenen Anschauung durch Reisen zweifellos ist, viel zu wenig. Dem fiinstlichen Kulturwald gleschen Alters aber haftet ein grosser Nachtheil an; man hat wenigstens in Europa diese Erfahrung aus bitteren Verlusten gesammelt, dass nemlich die Insektencalamititen in erdsserem Umfange und verderblicher aufzutreten pflegen, als sonst. Japan ist zwar in der gliicklichen Lage von solchen Catastrophen bisher verschont zu sein, ob aber die unter dem Drucke der wirtschaftlichen Notwendigkeit eintretende Umgestaltung seiner Waldungen nach innerer Verfassung und nach Holzarten nicht dieselben mit sich bringen werde, dafiir fehlt vorderhand jede Erfahrung. Wie erwahnt wiirde man gut daran thun, die Méglichkeit zu beriicksichtigen. Auch die Verlegung des Schwergewichtes nur nach ezzer Holzart hin, z. B. der Cryptomerie, hat, wenn zu uniform zur Geltung gebracht, manche recht bedenkliche Seite. Man sollte stets darnach streben Misch-bestinde zu erziehen, um bei einer ev. Anderung der Conjunktur des Absatzes nicht auf das Trockene gesetzt zu sein und cine bessere, intensivere Bodenausnutzung zu erzielen. Das stellenweise bemerkbare ginsliche Verdringen der Laubwaldungen durch Nadelholzer ist ein spater schwer wieder gut zu machender Fehler. Zweifellos ist ja der Nadelwald in Japan itberwiegend werthvoller und man wird deshalb Nichts dagegen einzuwenden haben, wenn der Laubwald praktisch in die Inferioritat gedrangt wird, aber ihn ganz vom Zukunftswalde auszuschliessen, wie man das an verschiedenen Plitzen bereits bemerken kann, ist ebenfalls verfehlt. Ich glanbe man gibt ihm am besten die Rolle cines Zwischen-und Unterstandsgliedes, dadurch wird die Bodenthiatigkeit besser gewahrt als durch die reinen Nadelholzbestande. Der reine Laub- wald, wo er allein angangig erscheint, wird zweifellos im hochwaldartigen Mittelwalde seine beste Wirtschaftsform haben. Auf gecignetem Boden und bei entsprechenden sonstigen Verhialtnissen ist auch sicher der Laubwald noch rentabel, sobald nur auf Erziehung eines schr werthvollen Materiales hingearbeitet wird. Schlechte Béden sind fiir Die zukinftige Bewirtschaftungsform des japanischen Waldes! 339 Laubwald als Hauptnutzholzwald aufzugeben, da hier die erwahnten Voraussetzungen hochwertiger Produktion nicht vorhanden sind. Die grosse Ausdehnung der Flache nach, nimmt in Zukunft der Laubwald in Japan sicher nicht mehr ein, und mit Recht, denn eine bessere Wirtschaft sucht Rentabilitét, aber auch waldbauliche Gesichtspunkte insbesondere Vielseitigkeit und Nachhaltigkeit mehr in Einklang zu bringen, das fuhrt dann unmittelbar zu ecmzschten Waldungen. Fir gemischten Wald ist aber die Grosskahlflachenverjiingung die wugcetguetste und muss ihr auch aus diesem Grunde der Werth, den sie in der Vergangenheit hatte, vollig abgesprochen werden. Ebenso unpassend ist aber der grosse Kahlschlag, der im Wesentlichen seine Heimath in-der Ebene hatte, fir gedirgige Lander, wo eben die Gefahrdung des Terranis durch die meteorischen Niederschlage in ganz anderer Art sich dussert. So viele verédete Plitze des Berglandes, welche heute in ihrem zerrissenen, den Abschwemmungswassern preisgegebenen Béden, die Zufuhrquelle der Geschiebe zu den verderblichen Wildbachen und Flissen darstellen, sind sicher grossentheils nur auf Kahlschlagwirtschaft im grossen Stile zuriickzufiihren. Wir hitten gewiss noch viel traurigere Bilder der Verwiistung des Bergterrains in Japan vor uns und eine noch viel drohendere Gefahr der Wildwasser, wenn nicht die beispiellose Reproduktionskraft des Bodens mit ihrer unendlich manigfaltigen Pflanzendecke hier zu Hilfe kame. Dass man aber damit auf die Dauer nicht auskommt, das beweisst zur Geniige eine Augenscheinnahme im Gebirge und die Hochwasserschaden- statistik. Die Extreme beriithren sich und nach diesem Grundsatze scheint man auch in Japan vorgehen zu wollen, denn nachdem zweifellos die Fehler der alten bis heute im Schwung befindlichen Grosskahischlag-damit kiinstlichen Verjiingungs- sowie Aussugshauungs- Politik sich nicht mehr laugnen lassen, will man sich zum direkten Gegentheil bekehren, namlich zur natiirlichen horst-und gruppenweisen V erjingung. Die Nutzung der Altholzbestiinde in langen Verjiingungszeitraumen und die Nachzucht der neuen Generation auf natiirlichem Wege in Gruppen und Horsten, unter Zuhilfenahme stellenweisen kiinstlichen Anbaues, entspricht ja auch der Bewegungsrichtung welche die moderne waldbauliche < 340 Dr. Hefele: Entwicklung in Europa cingeschlagen hat, somit cin Grund mehr, so meint man hier, sofort in radikalster Weise vorzugehen und zwei Treffer auf einmal zu erzielen, namlich besser zu wirtschaften und dann sich im Glanze des Bewusstseins zu sonnen, dass man auf der Hohe der Zeit ist. Dieser Ansicht wird man heute in Japan nicht selten begegnen und die Verfechter derselben fuhren als Argument an, dass diese Form der Bestandsnutzung und Ver- jungung nicht einmal vez sei, sondern lange Azer zu Lande geibt. Als Beweis wird dann eine vorhandene sogenannte Plenterwirtschaft theoretisch und bei Gelegenheit auch in praxi vorgezeigt. Wenn man eine vegellose Ausraubung des werthvollsten Materiales aus alten Bestanden und eine vél/7ge Gleichgiltigkeit tiber die zznzere Beschaffen- heit und den werthschaftlichen Werth der Nachzucht, eine Plenterwirtschaft heisst, dann allerdings haben die Anhinger dieser Richtung Recht. Der Hinweis auf die Natur, welche ja auch in ahnlicher Weise, wenn sie durch die Hand des Menschen nicht gestort wird, den Wald regenerirt hat, ist umso weniger gerechtfertigt, als sie eben den Wald xur als Selbstzweck weiter produzirt und verjiingt, ove sich um Zeit und Werth zu kiimmern. Die Frage nach Zeit u. Werth ist aber das Grundelement einer heutigen Waldwirtschaft, in der Beriicksichtigung dieser Faktoren liegt das Characteristicum des Begriffes ‘‘ Wirtschaft.’ Eines der Hauptziele einer wirklich 6konomischen Betriebes muss die Erreichung des desten Effektes unter Aufwand gcringster Mittel sein. Das technisch Vollkommenste nach den méglichen Verhaltnissen zu leisten, xzcht aber blos #berhaupt Production zu treiben, ist Aufgabe der modernen Forstmannes. NVatiirliche horst-und gruppenweise Verjingung, (Fehmelschagform) und bet Ausdchnung des Verpiingszettranmes auf den ganzen Umtrieb (Femel oder Plenterform) kann das Vorhandensein eines guten und entwicklungsfihigen Verwuchses oder die Schaffung schoner guter Verjiingungsgruppen nicht entbehren. Wo sie kritiklos zeden natiirlichen Vorwuchs beniitzt, auch wenn er mit 50 oder 70 Jahren héchstens 1 meter hoch ist, da verdient ein solches Verfahren den Namen einer Wirtschaft iiberhaupt zzcht mehr, es wird blanker Raubbau, dessen blenden-sollende geringe Kultur-Kosten nichts weiter als blanke Tauschung Unerfahrener bezwecken. Die zukiinftige Bewirtschaftungsform des japanischen Waldes! 341 Glaubt man sich aber damit beruhigen zu k6énnen, dass aus solchen schlechten, oben erwahnten Vorwiichsen ein gesunder geschlossener Wirtschaftswald sich entwickeln werde, so wiirde das die Wahrheit des Satzes voraussetzen, dass eine Jahrzehntelang wuterdriickte Baumpflanze bei Gewahrung von entsprechendem Lichtgenuss und Kronenfreiheit sich ebenso zum Hauptstamm entwickeln k6nne, wie eine gesunde, durch Druck zich degenerirte. Wie grundfalsch eine solche Annahme ist, zeigt schon das Fiasco des auf ihnlichen Voraussetzungen basierenden Borggreve’schen Durchforstungsverfahrens. Vollends in den héheren Regionen der Berge ist ein solcher Fehmelschlag oder Plenterbetrieb zwecklos. [ze/ Material kann nicht gewonnen werden und wo man nur mehr einzelne Stimme nutzen kann, da ist man meines Erachtens in die Region des Schutswaldes eingetreten oder ihr bedenklich zake gekommen und da muss, so lange die Waldungen der tieferen Lagen noch keiner richtigen Wirtschaft unterstellt sind, einfach vorerst die Hand davon gelassen werden. Man iibersieht aber auch an dex Platzen, wo aus natiirlichen Griinden horst und gruppenweise Natur- Verjiingung moéglich wire, ein paar Hauft- bedingungen einer solchen Wirtschaft vol/kommen. Sie setzt nimlich, wenn sie richtig betrieben werden will, einmal eine weztgehende Aufschliessung der Waldungen durch Wege etc. voraus und kann ferner auch nur von einem erfahrenen und geschulten Personal executiert werden. Ob diese conditiones sine qua non grundlegender Natur fiir Japan zutreffen oder nicht, dariiher glaube ich besser kein Wort verlieren zu sollen. Dinge, welche weitschauenden Blick, Umsicht und praktische lung der technisch herangebildeten Beamten sowie eine vorziigliche Dressur der Vollzugsorgane erfordern, kénnen meines Erachtens nicht wie eine Maschine einfach gekauft und in Gang gesetzt werden und im Decretieren ist man zweifellos weniger beengt, als in der Durchfitthrung in praxi. Die ungeheure Verantwortung aber, die darin liegt, dass man ein Nationalgut, wie den Wald, der, bei richtiger Behandlung, eine der besten Einnahm- quellen des Staates sein kann und in andern Liindern auch ist, zu gewagten Experimenten mit zweifellos sch/echtem Ausgang beniitzt, gibt doch wohl auch zu denken 342 Dr. Hefele: Die ganze Frage der Rentabilitét des Waldes in Fapan ist eine Frage der Entwicklung der Communicationslinien ! Welche Nutzung und Verjiingungsform ware demnach empfehlenswerth, bis dieser Entwicklung des Verkehrs durch Wege etc. mehr Rechung getragen ist ? Zweifellos die Kahlschlagsform in relativ schmalen Schligen, im Bergterrain der Hangrichtung folgend, also Sawmschlag mit nachfolgender kiinstlicher Verjingung durch Saat oder Pflanzung. Damit vermeiden wir ecinerseits die Gefahren des Grosskahlschlages, und bekommen mehr Material auf jedem gegebenen Hiebsplatze, als wie dies bei richtiger Fehmelschlag oder Fehmelwirtschaft méglich ist; davrauf muss aber gesehen werden, da eben die geringe Ausdehnung des Wegenetzes fiir manche substituirend eintretende Bringungsmethoden einen Aznreichenden Materialanfall als Bedingung fiir die Rentabilitat der Bringungsanlage erfordert. Die Saumkahlschlagform ist ferner technisch einfach und also mit dem vorhandenen Personale eher durchfiithrbar, sie ermédglicht die Bringung tiber die Schlagflache se/ds¢ in ungenirter Weise, und ist z. B. die giinstigste Abholzungsform in Berglandern, denen gute Hangaufschliessung durch Wege mangelt. Unter Zuhilfenahme entsprechenden Hiebswechsels kann auch die sichere Nachzucht von Bestanden jeder Holzart oder Mischung div. Holzarten sehr gut ausgefiihrt werden. Das m. E. nach den dermaligen Verhialtnissen auf lange Zeit hinaus in Japan Erretchbare ist damit gekennzeichnet. Wird einmal ein wohl- iiberlegtes richtiges Fundament, ein solider Unterbau, fiir die weitere Entwicklung durch Reorganisation der Waldwirtschaft Grundsatze mit zielbewustem Ersatz der diberstiirtsten, weder geniigend gekannten, noch tn threr Wirkung geniigend geschitsten Massnahmen gegeben, dann wird auch die Zeit sich nahern, wo der primaren /undamentalaufgabe, der Auf- schliessung der Waldungen durch Wege etc., ohne Schwierigkeit eine natirliche Verjingungsform europ. Stils wird folgen koénnen. Techniker, welche selbst practische Wirtschafter sind, miissen aber erst das Personal fiir diese hohe Auforderung mittlerweile schulen, was bei localer /angsamer Uberleitung (der Aufschliesung entsprechend) vom Saumschlag zur femelschlagweisen Verjiingung und suerst an kleinen Die zukiinftige Bewirtschaftungsform des japanischen Waldes! 343 Objecten probirt, bei consequenter richtiger Methode auch erreicht werden kann. Schwierigkeiten k6nnen nicht schrecken, wenn es das Wohl des Staates gilt, aber Hindernisse in waldwirtschaftlichen Dingen lassen sich nicht uberspringen, sondern nur langsam und schrittweise bestegen ! SS ec lll OOO . Wald und Wasserwirtschaft. VON Dr. Hefele. Kgl. bayr. Forstmeister u. Professor in Tokio. (Vortrag gehalten in der deutsch, Ost- Asiat, Gesellshaft fiir Natur u. Vilkerkunde in Tokio, “ Aus den Mittheilungen der D, O. A. G. f. N. u. V."’) Jeder Kulturfortschritt im Leben der Volker beruht nicht in letzter Linie auf einer intensiveren Ausniitzung der naturlichen Giiter, wie sie durch klimatische, oro-und hydrographische Verhaltnisse des von einem Volke beswohnten Landes dem Menschen von der géttlichen Vorsehung beschieden wurden. | Die notwendige Voraussetzung hiefiir ist zweifellos die Erkenntniss des Werthes derselben und die menschliche Einwirkung ist mitunter in ausgedehntem, ja staunenswerthem Masse mdglich, haufig findet sie aber auch ein schnelles Ende, je nachdem sie in Ubereinstimmung oder in Dissonanz mit den ‘“‘ewigen Gesetzen” der Natur ihre Wege ecinschlug. Das Grundfundament eines waren Fortschrittes muss also die Erforschung der Ursachen einer Erscheinung bilden und so sehen wir, dass mit der zunehemenden Durchleuchtung bisher dunkler Gebiete durch die alles durchdringenden Strahlen einer logisch vorgehenden exacten Wissenschaft, natirlich auch der Weg zum Ziele gerader und freier von irrefiihrenden Seitenpfaden wird. Gerade die Entwicklung der Naturwissenschaften in ihrem rapiden Gange hat der unanthaltsam vorwirts hastenden Zeit bei der Kulturmission die unschatzbarsten Dienste geleistet, wie ein Blick ins tigliche Leben ohne weiters lehrt. Dass manch alte liebgewohnte Vorurtheile oder Annahmen dabei zu Fall kommen und manche trotzige Burg einer Scheinwahrheit ihre 346 Dr. Hefele: Jahrhunderte alten Grundfesten untergraben findet, ist nur natirlich, aber ungern trennt sich der Mensch von einer durch Alter geheiligten Uberlieferung. Die Besprechung des vorliegenden Themas wird zeigen, dass es in seinem Gegenstande zu einem der heissurmstrittenen gehdért, da eben eine allseitig einwandlose Lésung der damit vorknipften Fragen noch weitererer exacter Forschungen bedarf, wenn auch das Endziel in groben Umrissen schon jetzt £/ar vor Augen steht. Wie aber die Ungleichartigkeit von Kraften allein eine heilsame Storung des starren Gleichgewichtes veranlasst und damit als Grundelement der Bewegung im Sinne der Bildung neuer Phasen betrachtet werden muss, so kann auch nur durch den Widerstreit des Meinungen, basirt auf griindlichen Untersuchungen, die Vorbedingung geschaffen werden ftir das Aufsteigen des Phénix ‘“ Wahrheit und Fortschritt”? aus der Asche der im Kampfe zerstorten Vorurtheile. Der Grund warum bei der eigentlich beabsichtigten Ausprache tber die “ Wasserwirtschaft”? und deren Werth fur ein Land, der “ Wald” als eleichberechtigt, ja voranstehend genannt wird, liegt in der Abhangigkeit der Letzteren zum grossen Theile von den Verhiltnissen des Ersteren. Im Wasser ist den Menschen eine Naturgabe von der Schopfung verliehen, welche nicht nur in fundamentalster Weise das organische Leben auf dem Erdball beinflusst, sondern in weiterer Ausgestaltung seiner Benutzung dem schwachen Geschlechte der Erdenbewohner jene Riesenkrafte leiht, welche zur Durchfihrung der immensen Plane seiner nimmer rastenden geistigen Capacitat eines der gréssten Hilfsmittel darstellt. Gewissermassen schon der blosse Anblick der Weltkugel scheint uns die Bedeutung des Wassers auf unserem Planeten ziemlich eindringlich zum Bewusstsein bringen zu wollen, denn 3/4 der Oberflache ist allein von den Meeren eingenommen. Die Bedeutung dieser internationalen Handelsstrasse der Volker wiaichst mit jedem Tage und die Ausgestaltung der Verkehrs- mittel durch die Technik macht sozusagen die Welt nach und nach kleiner unddie Mission der Kulturstasten grésser und grésser, bringt das Zzel der geistigen und sittlichen Hebung der Volker der Erde naher und naher. Um aber in diesem Austausch der realen und geistigen Produkte einen Wald und Wasserwirtschaft. 347 durch historische Vergangenheit namentlich aber den verliehenen Talenten entsprechenden Platz im Rathe der Volker fiir den Verkehr iiber die Meer zu behaupten, ist die Entwicklung der Produktion im Innern der Linder eine bindende Voraussetzung. Und hier kommen die Binnengewisser die Seen, die Fliisse und Biche zur Geltung. Sie bilden den Kernpunkt der Giitererzeugung sei es direct durch Fructificirung der im natiirlichen Boden vorhandenen Niahrstoffe fiir das Wachsthum von Feldfriichten, wie im Agrarstaat oder indirect, als Lieferanten motorischer Kraft fir die Maschinen des Industriestaates. Stets aber ist die Erzielung einer segensreichen Wirkung gekniipft an cin gewisses Ebenmaas, eine gewisse continuirliche Bestindigkeit, nirgends sind Extreme nach dem Zuviel wie Zuwenig von solch einschneidenden Folgen begleitet, wic hier. Die Austattung eines Landes mit zahlreichen guten Fliissen und Wasserliufen gibt einen guten Masstab der Beurtheilung seiner Entwick- lungsfahigkeit gegeniiber cinem anderen Lande bei sonst gleichen Beding- ungen. Je grésser die Stréme, je tiefer dieselben sind, je leichter und je weiter sie vom Meere landeinwirts befahren werden kénnen mit grossen Schiffen, desto werthvollere Verkehrsmittel stellen sic dar und, wenn es auch eine Zeitlang schien, als ob ihre Bedeutung durch die Eisenbahnen in den Hintergrund gedringt wiirde, so hat eben die neueste Zeit mit ihren Riesenprojekten, z. B. dem Mittellandkanal in Deutschland bewiesen. wie gerade bei stark entwickeltem Verkehr und wirtschaftlichem Aufschwung die Wasserstrasse der grossen Fliisse u. Kaniile wegen ihrer hohen Leistungs- fihigkeit und .Billigkeit, speciell bei im Eigenwerth niedrigen Giitern, nicht entbehrt werden kann. Wo Mangel an natiirlichen Wasserwegen herrscht, wird sich das EKisenbahnnetz bei ‘giinstiger Aussicht der Produktion verdichten aber immer wird es auch mit dem Nachtheile grosserer Kosten behaftet sein Die gute Benutzbarkeit der Wasserstrassen hingt jedoch nicht zum geringsten Masse von regelmiissigen IWoasserstdénden und méglichst w«z- verdnderlichen Sohlenverhiltnissen ab. Da es bei den grossen Strémen vor allem der Oberlauf und die Seitenfliisse sind, welche die giinstigen oder uugiinstigen Verinderungen veranlassen, und cine Einflussnahme von 348 Dr. Hefele: Seiten des Menschen im Grossen Ganzen /der cinsetzen muss, so ereiebt sich von selbst deren grosse Wichtigkeit im Systeme der Wasserwirtschaft. Die im Nachfolgenden betrachteten wertschaftlichen Vortheile der kleineren Fliisse und Wasserlaufe werden von den grossex Verkehrstrémen, bis zu cinem ausgiebigen Grade getheilt, so dass thre gemeinschaftliche Anfihrung zweckmiassig erscheint. Wahrend bei den kleineren Flissen und Wasserliufen der Werth als grosses Verkehrsmittel des Handels wegfallt, leisten alle Wasserlaufe fur Landwirtschaft und die /udustric nach mehrfachen Richtungen Unersetz- liches. Die Landwirtschaft kann sie nicht entbehren im Sinne: 1) der Entwasserung. 2). Zum Zwecke der Bewadsserung © * & Fructification des Bodens, wahrend die vom Gewerbe u. der Industrie so sehr gesuchte O2//z¢¢ Betriebskraft und dic Méglichkeit eines billigen Lokaltransportes fur Rohprodukte, bei ¢7déssercr Entwicklung des landwirtschaftlichen Betriebes selbstverstandlich von demselben durch die Zuhilfenahme der Maschinen etc. im Sinne der gleichen Vortheile, wie sie die industrielle Dhatigikent sucht, ebenfalls getheilt wird. Die Lutwsdéserung des Landes durch die Iliisse ist gewissermassen ihre natirlichste Aufgabe, denn das metcorisch niederstro6mende Wasser muss diesen Ausweg haben, wenn nicht cin Hinderniss der Kultur und damit der Bewohnbarkeit eines Landes durch Versumpfung geschaffen werden soll. Die Adfussgrésscn der Fliisse sind von verschiedenen Umstinden bedingt. In erster Linie von der Jlenge der Jallenden Niederschlige in ihrem Einzugsgebiete. Wo diese in einem Lande nur gerizg sind, wird man vergeblich nach Wasserliufen von einiger Bedeutung suchen. In Aden z. B. fallt durchschnittlich alle drei Jahre cinmal ausgiebiger Regen’ und die Vorkehrungen durch kiinstlichen Ausban natiirlicher Tels- becken im Gebirge als Wasserreservoire, um nur das nothige Trinkwasser fir dic spiarliche Bevélkerung zu erhalten, sind weltbekannt. Der Missisipi der gewaltigste Strom Nordamerikas mit dem ungeheuren Einzugs- ecbiete von 3,150,000 |. Km. fihrt dagegen von den pro Jahr ca 90,000. Millionen engl-Cubicfuss = betragenden Niederschligen auf diesem Wald und Wasserwirtschaft. 349 Territorium, ca 19,500 Millionen Cubicfuss dem Meere zu. (25% der Nicder- schlige.) Die Riesenstr6me Siidamerikas (Amazonen & Laplata-Stom) und des Ob, des gréssten asiatischen Stromes in Russland, des. Yantzckiang in China lassen in ihren gewaltigen Wassermassen einen Riickschluss auf dic Menge der Nicderschlige in threm Einzugsgebiete sowie auf dessen Grésse zu. Freilich ist die Vertheilung der Niederschlagsmengen in den angefiihrten Stromeinzugsgebieten selbst cine umso ungleichmiissigere, je klimatisch und orographisch verschiedenere Regionen umschlossen bzw. drainirt werden. Das Verhdltniss zwischen Niederschlagsmenge und Abfuhrimassen ist keineswegs cin koustantes sondern, wie leicht erklarlich, von den variablen Verhiltnissen des Bodens nach natiirlicher Form, piysikalischer Beschaffenkeit etc. abhiingig und am mezsten bedingt durch das Feh/en oder Vorhandensein einer Pflanzendecke und deren Zasammensetsung. Bei der Besprechung des Einflusses des Waldes werde ich hierauf ansfiihrlicher zuriickkommen. Was dte drainierinde Thitigkeit der Fliisse stért, wird naturnot- wendiger Weise auch dic giinstige Wzrkung der Drainage becintriichtigen. So sehen wir denn iiberall, namentlich in den mittleren und unteren Flussliufen mit den geringen Gefallen, woselbst in der Regel dic Ebene das Charasteristicum der anstossenden Landschaft bilden wird, durch Verwilderung der Fliisse, also bei mangelndem Fingreifen des Menschen Uberschwemmungen, Ejisstopfungen beim Eisgang, Neigung zur Bettverle- gung, Versandung cte, in allen Fiallen aber cine starke Verminderung der Abfiithrung des Wassers ecintreten. Es muss also die Sorge einer Landes- verwaltung auf cine Correction und Uberiwwachung ihrer Wasserliufe ecrichtet sein, nattirliche Abflusshindernisse miissen bescitigt und kiinst- liche Einbauten auf ihre Wirkung in etwa nachtheiligem Sinne zuvor wohl eepriift werden. Die zweite wichtige Aufgabe des Flussliiufe niimlich der “ etwdsse- rung” des Kulturlands hat cine weitaus ¢réssere Beachtung von Scite des Menschen gefunden und wird dies Verhiiltniss wohl auch in Zukunft beste- hen; die Méglichkeit der Fruktificirung ungeheurer Flichen ist in viclen Fallen nichts weiter als cine Frage der Einleitung von geniigend Wasser 350 Dr. Hefele: um mineralisch tauglichen Boden durch die Zuftihrung der Pflanze néthigen, bisher fehlenden, Feuchtigkeit, in ertragsreiche Kultur umzuwandeln. Das grésste Project der Neuzeit dieser Art, dirften die Nilsperr- dimme bei Assuan bezw. Szuvt im oberen resp. mittleren Aegypten sein, welche in Verbindung mit den bereits bestehenden bei Kairo eine systematische Ausniitzung der Nilfluth im Frijahre in grossartiger Weise bezwecken. Durch Sperrdimme sollen Stauseen geschaffen werden, deren Wasser nach Bedarf zur Abgabe kommt und die Bodenbenutzung in ausgedehntem Masstabe, namentlich ftir héher gelegene Liindereien, welche mit den bisherigen Staumitteln Wasser nicht erhalten konnten, garantirt. Den riesigen Kosten von 2 Mill Pfund stehen dauernde Erhéhung des Nationalvermégens, der Steuerkraft des Landes und Einnahmen aus dem zunehmenden Verkehre in tiberlegener Grésse gegentiber. Bereits ertragsfahiger Boden wird in seinem Ertrage durch dauernde Bewdsserung gesteigert, das wird namentlich dort zur Geltung kommen, wo sonstige giinstige klimatische Faktoren noch unterstitzend cingreifen. In Europa hat Frankreich den 31. Theil, Italien den zwangigsten Theil seiner Landfliche zur Bewisserung eingerichtet, wahrend in unserem Heimathlande Deutschtand diese Ziffer nicht erreicht ist. Je mehr bei wachsender Bevélkerung und nicht beliebig vermehr-bezw. cultivirbarem Boden geringere Boden der Kultur unterworfen werden, desto mehr wird auch das billige Diingemittel des Wassers in Anspruch genommen werden. Im Allgemeinen sind es auch hier wiederum die kleineren u. mittleren Wasserliiufe welche ftir solche Zwecke in Betracht konnen, denn die grossen liisse bediirfen zu ihrer Nutzbarmachung kostpieliger Anlagen, welchen nicht immer die entsprechenden Gegenwerthe gegeniiberstehen. Was endlich den Werth von Wasserlaufen fiir Gewerbe und Industrien betrifft, so ftreben Gewerbe, wie industrieller Betrieb im engeren Sinne nach Ersatz der menschlichen Arbeitskraft durch den J/otor, dessen Antrieb von einer méglichst billigen, leicht erhiltlichen und nachhaltigen Kraftquelle i. e. dem Wasser erfolgen soll. Auch diesen Auforderungen entsprechen die geringe- ren Wasserliiufe besser im Durchschnitt als grosse Stréme, da letztere wohl immense Krifte liefern aber auch enorme Anlagen zur Realisirung derselben Wald und Wasserwirtschaft. 3 wt — erfordern. (Nutzbarmachung des Niagarafalls in Amerika.) Die Bedeutung der Kraft des Wassers als Antriebsmittel nimmt zu, je mehr der Grossbetrieb decentralisirt wird, was hinsichtlich einer Anzahl Industrieen z. B. Holzindustrieen schon der Fall ist. Die Moglichkeit der electrischen Kraftiibertragung ist zweifellos als die Hinwegraumung cines Hindernisses fiir die Nutzbarmachung mancher Wasserkraft zu betrachten. Die Wasserleitungen der grossen Stadte greifen endlich zuriick bis in die Quellengebiete, also den Anfang aller Wasserliiufe, und die Wirkungen derselben in sanitarer Hinsicht bedtirfen keiner niiheren Begriinduneg. Den Seen Kommt jene Bedeutung wie den Fliissen & Biaichen weniger zu, sie sind mehr lokalisirt, es fehlt ihnen die Modulation der Liingenent- wicklung, obwohl sie unter Umstinden im gleichen Sinne wie laufende Wasser nutzbar gemacht werden kénnen. . Diesen segensrcichen Wirkungen des Wassers der Binnenfliisse etc. von denen eine einzige geniigt, um ihnen die Kigenschaft der Unentbehrlickeit zu verleihen, stehen Verheerungen und Katastrophen gegeniiber, welche in manchen Lindern durch ihre haufige Wiederkehr die Bewohnbarkeit und Cultur weiter Jandesstrecken annullierten und ganze Gegenden der Verédung preisgaben. Je mehr die V6lker der Erde dieselbe occupieren, je dichter sie auf gegebenem Raume sich concentriren, desto mehr wird man sich der Bekiimpfung des den Boden bedrohenden Ungeheuers widmen. Wohl wird es niemals méglich sein, cigentliche Katastropfen gréssten stils, welche in Stérungen der Atmosphiire cte. ihre Ursache haben oder einer Verinderung von bisherigen Gleichgewichtsfaktoren im Weltsysteme aur Last fallen mégen (man denke nur an die Theorie der allmihligen Abkiihlung der Erde bis zur Vereisung, an den Einfluss der Sonnenflecken etc.) abzuwenden, aber bis zu einem gewissen Grade besitzt der denkende Mensch Mittel, um sich zu schiitzen und zu wehren gegen den Vernichtungs- kampf, den ungeziigelte Naturkriifte gegen ihn fithren, u. wenn auch nach nach dem Dichterworte “denn die Elemente hassen das Gebild von Menschenhand,” alles irdische Streben citel zu sein scheint, so diirfen wir doch die Hiinde nicht in den Schooss legen und miissen das Jenschen- 352 Dr. Hefele: mogliche zu erreichen streben. Man hat’sich in neuerer Zeit nicht mehr begnigt mit der Bekimpfung der Folgeerscheinung, dem Hochwasscr als solchem, sondern ist der Frage nach den Ursachen nahergeriickt. Hier findet man nun in der Litteratur den vom Volksglauben lingst als Ursache bezeichneten Hauptfaktor, nemlich den Zustand des Ursprungsgebietes der ePiltsse hinsichtlich seiner Pflanzendecke, und in der Hauptsache ist der “ Wald” damit gemeint, in Herz und Nieren gepriift auf den Zusammenhang mit den verheerenden Wirkungen der Wasser in den Fliissen. Wir sind damit unwillkirlich in jenes Gebiet der Wasserlaufe cingetreten, das uns Forstleute naturgemiiss am meisten interessirt, in den Odcrlauf, das aber auch, wie ich darzuthun hoffe, die meiste Aussicht auf erfolgreiche Becinflussung der heute so brennenden “Wasserfrage” gibt. Ich beschranke mich daher im grossen Ganzen auf dic Besprechung der mdéglichen Kinwirkungen auf die Wasserliufe, im /zstehungs-Gebicte, welches wohl meistens im Gebirge zu suchen ist und auf die Bekiimpfung der Ubel in den obcren im Gebirge gelegenen Theile der Fliisse, wo die gvossen Gefalle und insbesondere die Geschichecfiihrung die meisten Schaden verursachen, wihrend im mittleren und unteren Laufe der Str6me mehr die Menge der zugefihrten Wassermassen verderblich wird. Der erstere Theil besitzt vieleicht nicht immer jene raumliche Ausdeh- nune der Schiiden, wie sic die Niederungen der Miindungsgebiete und die Mittellaufe der Fliisse aufweisen, aber an /xtensztét der Verwiistung kommt kein anderer Theil ihm gleich. Hier ist das Terrain der Wildwasser, der “ [Waldbdche” und ihrer Vermuhrungen und jeder Erfolg der hier errungen wird, kommt den Niederungen der bene zu Gute und zwar in potenzirtem Maase. rigt man zunichst, che man den Wert der “ Bewaldung” untersucht, ob und in wéewert cine Pflanzendecke im Ursprungsgebiete ttberhaupt einen KKinfluss ausiibt auf das Regime der Wasser, so kommt man auf Grund der Untersuchungen des bayrischen Prof. Ebermayer, des leider zu friih verstorbenen It. Wollny in Miinchen u. Anderer zu dem Schlusse, dass jede Pflanzendecke (Gras, /ara) etnen Vorsug gegeniiber kahlem Gebirge bedeutet. Wald nnd Wasserwirtschaft. 2 wi > Unschwer ist cinzusehen, dass jeder Bodeniiberzug perennirender Gewiachse einen gewissen Ausgleich in der Wasserfithrung hervorruft, indem die ober-wie unterirdische Wasserableitung verz6gert wird, was in der Gesammtwirkung gleichmiéssigerer Wasserstinde scinen — schliesslichen Ausdruck findet. Eine Verlangsamung der oberirdischen Abfuhr bedeutet cben cin besseres Lznsickeren der meteorischen Wasser in den durch dic Pflanzen- wurzeln physikalisch ginstiger d. h. lockerer gemachten Boden, wodurch Durchléssigkeit und Aufnahmsfihigkcit sich steigern. Fir die wichtige Frage der Quellenbildung- u. Erhaltung an den Hingen der Gebirge kommt in erster Linie das langere Verweilen des oberirdischen Wassers auf der betr. Flache in Betracht, denn je mehr davon in den Boden einsickert, desto desser wird die Sfetsung der Ouellen erfolgen, die ja nichts weiter sind, als tiefer zu Tage tretendes Sickerwasser boherer Lagen. Sicht man von “ Wald” vorerst ab, so ist im Ubrigen bei der Héhen-Lage der Linzugsgebicte meist nur cine Buschvegctation nud in der Hauptsache “ Gras” als Bodendecke vorhanden. Gras erfiillt aber den Zweck der Wasserverlangsannung u. der Verhiit- ung des Terrainangriffes nur sehr mangelhaft, denn cin dichter Grassfilz ist nicht genugend durchlassig und bringt namentlich in steilen Lagen cin rapides oberfiichliches Abfltessen mit sich, ist also zweifelios fiir Gleich- missigkeit der Wasserfiithrnng der Fliisse und fiir Quellen von énferiorem Werthe, wenn auch besser als nakter Boden. Lockere Grasdecken (Hara sind aber keineswegs gegen den Angriff des Wassers gesichert, wie der Augenschein in Japan Iehrt. Als man in Frankreich 1864 hoffte, cine Beruhigung der gefihrdeten Flachen im Berglande durch BERASUNG der vou Wald entbléssten, durch die meteorischen Wasser in Bewegung gerathenen, und dic Wildbiiche mit Schutt fiillenden Berghiinge zu erzielen, da erwiesen sich die daran gekniipften Hoffnungen als triigerisch und es blicb nichts anderes iibrig als zur dufforstung zu greifen. Ich denke, das ist cin deutlicher Fingerzeig fiir die Zukunft der wir z. B. in Japan entgegenst suern, wenn die riicksichtlose Verschlechterung der //era in steilen Lagen weiter gefiihrt wird, abgeschen davon, dass, wie erwihnt, der Graswuchs an sich, auch wenn er den 354 Dr. Hefele: Boden im conereten Falle halt, dennoch fiir eine grossere Regelmiassigkeit der Wasserstinde im Sinne der Verlangsamung des Abflusses des Atmospharilien und somit der seztlichen Verthetlung der Wassermassen starker Regen cte. zichts Hervorrgendcs \eistet. In Japan muss aber der Frage siwischen dem Zusammenhange der Bedeckung der Gebirge mit entsprechender Vegetation und den Wasser- verhdltnissen cine erhéhte Aufmerksamkeit geschenkt werden aus sez Griinden. 1) Infolge der schmalen Ausformung des Landes mit seinen der Langsrichtung folgenden ziemlich hohen Gebirgsziigen ist die Langenentwicklung der meisten Fliisse eine sehr geriumge, es herrscht das bei grosser Lange von Fliissen in anderen Landern auf den oberen Lauf beschrankte s¢ez/e Gefaille und der “ Weldbach- artige Charakter” vor, ja verlasst sie zumeist zzchkt bis zu ihrer Einmiindung ins Meer, wic man an den Verhiltnissen einer Anzahl derselben Fujigawa, Oigawa, Kisogawa cte. cte. beobachten kann. 2) Die Ausformung der Hinge im Gebirge ist in diesem volkanischen Lande eine extra sfez/c ; die erodierende Thitigkeit des auf diese Art rascher wie sonstwo zum Abflusse kommenden Niederschlags- wassers wird im wrzgiinstigen Sinne noch unterstitzt durch die - durchgingig sehr weiche Verwitterungsdecke alter Schiefer etc, welche dem Grundgesteine aufliegt. Wenn unter der bisherigen Benutzung als Hara ein grosser Theile des Berglandes in Japan scheinbar noch zxcht so sichtbar gelitten hat, als man nach dem Gesagten vermuthen kénnte, so ist zu bedenken, dass wir fur de dem Durchschnittsreisenden zu Augen kommenden niedrigeren Vorberge eine durch das Klima dedingte ausserordentliche Regenerationskraft der Vegetation haben. Diese Kommt aber fiir hohe, kihlere Lagen nicht mehr so sehr in Betracht. Deutlich erkennbar sind auch schon fiir den Laien die umfangreichen Terrainzerst6érungen im Bergland des mittleren und siidlichen Japan. Von Kobe bis Shimonoseki priisentiren sich die Berge im ra@udigen Zustande d. h. es blinkt der rothgelbe, nakte Grund der Hinge durch die zerstorte Alara und den Wald. Wer aber erst einen Blick in das /nunere Wald und Wasserwirtschaft. OW wn wr gethan hat, dem ist kein Zweifel tiber die unheimliche Thitigkeit des Rachegeistes des untergegangenen Waldes. Es bleibt nun zu untersuchen, in wie weit der “ Wald” als der Ausdruck der h6chsten organischen Gestaltung der Bodendecke der Erde einen Einfluss auf die Wasserverhdlinisse eines Landes hat. Diese Frage lasst sich nicht beantworten, ohne dass man die Sache in zwei Abschnitte theilt, namlich in die sog Il. “Waldklimafrage” d.h. den Zusammenhang des Waldes mit dem Klima eines Landes im Allgemeinen oder auf concreten Ortlichkeiten und, Il. Die Wirkung des Waldes, u. zwar des Bergwaldes in erster Linie, auf Regelung des Wasserabflusses u. Geschiebefithrung. Der Glaube an eine heilsame Wirkung des Waldes hinsichtlich einer Verbesserung des Klimas durch Milderung der Temperaturextreme, Ver- mehrung von Niederschlagen, Verminderung der Hagelgefahr etc., kurz als klimatischen Factor ist ein sehr alter, nichtsdestoweniger jedoch jetzt als eines der unter dem Ansturm der exacten wissenschaftlichen Forschung gefallenen Vorurtheile zu betrachten. Man hat auf die Trockenheit resp. Regenarmuth der Mittelmeerlinder im Zusammenhange mit ihrem geringen Waldreichthum hingewiesen, das Beispiel von Italien & Griechenland angefiihrt, wo grosse Linderstrecken durch die mangelnden Niederschlige zu :Wiisten geworden seien, als man den schiitzenden Wald zerstért hatte und kein Geringerer als ein Alexander von Humboldt hat auf die Abnahme der Regenmenge und der Luftfeuchtigkeit durch die Zerstérungen der Waldungen aufinerksam gemacht. Die reiche Zahl der Anhiinger dieser Anschauungen wies die hervorragendsten Namen der Wissenschaft auf, aber als man der Sache durch die meteorologischen Stationen und deren Beobachtungsresultate nahertrat, namentlich aber durch die Ergebnisse der Untersuchungen iiber die Existenzbedingungen des Waldes, da gelangte zu dem anfangs Staunen er- regenden Resultate, dass nicht der Wald einen Einfluss auf dass A7éma habe, sondern, dass die Baum- und Waldvegetation vollstiindig von den klimatischen Verhiltnissen einer Gegend abhiingig sei, so dass die bisher G1 iiblichen obenerwahnten Annahmen nur theilweise richtig sind. 356 Dr. Hefele: Der Wald ist niamlich weder natiirlich vorhanden noch jemals kiinstlich bei sonst giinstigsten Verhaltnissen dauernd begriindbar, wenn z. B. wahrend der Hauptvegetationszeit (in der nérdl. gemassigten Zone Mai—August) nicht in minimo 50 millimeter Niederschlag auf die betreffende Gegend fallen. In Aden 3 B. wird niemals eine Baumvegetation ohne kiinsthche Bewdsserung denkbar sein; die Ebene Californiens verdankt ihren Ruf als Obstkammer Amerikas nur den kiinstlichen Bewadsserungsanlagen; die ersten Ansiedler trafen kezze Baume an, da eben das xéthige Minimum an Wasser dem Boden bei sonst eminent klimatischen Vorziigen nicht geboten wurde. Baumwuchs ist aber trotz geniigender Bodenfeuchtigkeit (:Niederschlige:) ebensowentg denkbar, wenn die notwendige relative Luftfeuchtigkeit von mindestens 50% wihrend der Vegetationszeit nicht vorhanden ist. Da wir auf diese zweite Bedingung kaum je Einfluss gewinnen, so ist das Bestreben der Griindung von Wald adsolut aussichtslos in diesem Falle, mag die Bodenfeuchtigkeit noch so reichlich vorhanden sein. Beweise fiir diese Thesen finden sich durch die ganze Welt in prignantem Ausdrucke. Die Grasprarien im Centrum Nordamerikas und die Pampas Siidamerikas und Australiens, die Wiisten Afrikas, Aszens, (Gobi in China) sie kénnen nie und nimmer durch des Menschen Hand auch mit Aufwand aller Mittel, Baum oder Waldvegetation tragen, da die Luftfeuchtigkeit in geniigender Menge nicht gegeben werden kann. Das Sinken der relat. Luftfeuchtigkeit unter 509) wahrend der Vegeta- tionszeit kann nur von Gras oder Staudengewichsen ertragen werden, (Prarien Amerikas n. Australiens) wahrend ein Herabgehen der Luftfeuchtigkeit unter 40% rel. Feuchtigkeit und eine Niederschlagsmenge unter 20 m/m wahrend der Haupt-Vegetationsmonate die vegetationslosen Wiisten (Asiens u. Afri- kas) zur Folge hat. Nur auf den Grenzgebieten von Pririe und Wald ist anscheinend eine geringe Verschiebung in Sinne der Ausdehnung des Waldes méglich, aber bei genauerer Untersuchung zeigt sich immer, dass man ehemaligen Waldgrund vor sich hat, der durch irgend eine gewaltsame Einwirkung zu Grunde ging, was beispielsweise in Amerika im ausgiebigsten Massstabe bemerkt werden kann, indem es durch die Hilfe des Feuers leider a ee Wald und Wasserwirtschaft. 35 NI gelang, den Wald auf einer direkt nord-siidlich streichenden Linie, parallell dem Missisippilaufe zuriickzudriangen u. zwar um volle 10 Lingengrade (100°-go° westl. L.), eine gewiss nicht zu unterschitzende Leistung ! Dass man fzerzsulande in z. B. Hokkaido durch das schonungslose und unvernitinftige Anziinden der Bodeniiberziige des Waldes in der Nihe der der Ackerkultur gewonnenen Thiler das amerikanische Beispiel, wenn auch im schwicherem Masstabe, aber mit einer fiir den Nadelwald ebenso tétlichen Wirkung wiederholt, habe ich schon in einem fritheren Vortrage erwihnt. Weder die hohen Jemperaturgrade im positiven Sinne, welche in den sonnigen Wuisten Afrikas herrschen, schliessen bei gegebenen Beding- ungen der Feuchtigkeit den Wald aus (Oasen), noch Wintertemperaturen von—30 u. 40° C, wie man sie zz America besbachtet hat. Ein grosser Theil Sibiriens mit seinen Wintertemperaturen von oft- 45° C. ist voll der reichsten Waldschitze. Erforderniss ist einzig und allein eine mittl Temperatur von +72 bis rg? C. wahrend der Hauptvegetationszeit. Wird diese nicht erreicht (+8 bis 72° C), so sinkt der Wald zur Stauden u. Buschform herab. Tritt eden Monat des Jahres Frost ein,so ist Vegetationslosigkeit die Folge. Jnnerhalb dieser natirlichen Existenzfaktoren gliedert sich der Wald nach form und Zusammensetsung aus Arten, den lokalen klimatischen Verhdaltnissen entsprechend, und da die jeweils nérdlicher bezw. siidlicher vom Aequator gelegene Zone wut einer néher gelegenen, aber tn grésserer Elevation befindlichen correspondirt, so ist leicht zu erkennen, wie nach relativ einfachen Gesetzen die Bedeckung der Continente durch Waldfora zu denken ist. Wenn dem Walde auch ein genereller Einfluss auf das Klima iber ganze Flussgebiete und Landstriche oder Continente adgesprochen werden muss, so darf doch die Méglichkeit einer Fznwirkung aut klimatische Verhiltnisse im Sinne /oca/er Modulation ebenfalls nicht iibersehen werden. Ein sogenanntes “ Lokalklima”’ kann, iihnlich wie an einem grésseren See besondere Luftstr6mungen herrschen, vom Walde éetnflusst werden und wird sich natiirlich dies umso mehr geltend machen, um je gréssere Waldflichen es sich handelt. Die Klarlegung aller dieser Verhiltnisse muss den eingeleiteten Versuchen iiberlassen bleiben und wenn der Kampf der Meinungen dariiber 358 Dr. Hefele: auch in den berufenen Kreisen noch heftig tobt, an dem entscheidenden Hauptmomente, dass der Wald klimatische Katastrophen zzch¢ verhindern kann und keznen Einfluss auf Vermehrung u. Vertheilung der Niederschlige etc. hat, ist mit allen Consequengen kezz Zweifel. Der Hauptwerth der Waldbestsockung fir geregeltere Wasserstande der fliessenden Gewiasser und damit ftir die Verhinderung der zerst6renden und verderblichen Hochwasser oder fiir Erhéhung der fiir Kultur und Industrie gleich misslichen, zu ntedrigen Wasserstinde, liegt auf einem anderen Gebiete. Bewaldung erhéht namlich unter Bedingungen einerseits die Szcker- wassermenge, was von hervorragendem Einfluss auf die Spetsung der Quellen ist, und stellt anderseits von allen Bodendecken das bedeuteudste mechanische Hinderniss gegen die Abschwemmung der Bodens u. Abrut- schung der Scheedecke bet Lawinenbildung etc dar. Die erste Behauptung der Erhéhung der Sickerwassermenge, also der QOuellenvermehrung, kann in ihrem wollen Umfange nach dem heutigen Stande der Wissenschaft zzcht mehr aufrechterhalten werden und damit fallt wiederum eine seit alten Zeiten gehegte Ansicht. Wie weit sie noch Geltung hat, werden wir sehen. Die Untersuchungen von Ebermayer in Miinchen und Ototzkij’s in Petersburg haben den Nachweis geliefert, dass der Untergrund unter Wald in der Edene unter allem Umstinden a&rmer an Feuchtigkeit ist, als auf freiem Lande, dass also dem Walde der Ebene ein Einfluss auf Vermehrung der Bodenwasser im Sinne etwa der Erhéhung des Grundwasserspiegels u. der besseren Speisung von Quelle zzcht zukommt. Das scheint im ersten Augenblicke stark zu contrastiren mit Ergebnissen von Versuchen, welche zahlreiche Autorititen hinsichleich der Wasser- Absorption und Retention des Waldes und namentlich seiner Strexu angestellt haben. Durch die Uberdeckung des Bodens mit Wald wird in erster Linie dessen oberflichliche Verkrustung, teilweise erzeugt durch mechanische Wirkung (Festschlagen durch Regentropfen) verhindert, ferner, als eine Folge der durch die Humuserzeugung der Strendecke bewirkten besseren Krimmelung und Lockerung der Struktur des Bodens bei sonst gleicher mineralischer Beschaffenheit die Au/nahmsfihigkeit fir meteorische Nieder- Wald nnd Wasserwirtschaft. 359 schlige erhéht. Die herrschenden xiederen Temperaturen im Walde wahrend der hauptsiachlich wichtigen wairmeren Jahreszeit,—im Winter ist der Boden an sich durch Frost undurchlissig oder die Bedeckung mit Schnee als eine momentan latente Wasserquelle zu betrachten,—und die dazu kommende aus der Transpiration der Pflanzen resultierende gréssere Feuchtigkeit der Luft (3-10% im Mittel mehr gegeniiber dem Freilande) wirken im Sinne einer Vermehrung der Wassermenge des Bodens indirekt, indem dadurch die Ver- dunstung des atmosphirisch zu Boden gelangten Wassers verhindert wird. Hine gute, in richtigem Zersetzungsgrade befindliche Streudecke, welche im wohlgepflegten Walde niemals fehlt, vermindert die Verdunstung des der Bodenfeuchtigkeit nochmals ganz bedeutend, so dass die Verdunstung im Walde etwa nur 20% von jener einer Freilandsfliche betriagt. Diesen giinstigen Faktoren gegeniiber, welche die S7ckerwassermenge experimentell nachgewiesenermassen um nicht weniger als 24% fiir den _ streubedeckten Waldboden gegeniiber dem waldlosen erhéhen, steht nun der aufstockende Holzbestand bis zu einem gewissen Grade fezndlich gegeniiber. In erster Linie werden von dem atmosphirischen Niederschlagswasser Ca. 25% desselben durch die Baumkronen absorbiert, welche tiberhaupt z7cht an den Boden gelangen kénnen und ist diese Ziffer natiirlich nach Waldzustand Holzart, Alter etc. einer gewissen Veriainderung unterliegend. In dichten Fichtenbestiinden werden die Absorptionsprocente der Kronen bis zu 40 und 45% ansteigen und in lichten Lanbwaldbestinden ungekchrt niedrige Betrige aufweisen. Das bisher nicht geniigend in Rechnung gezogene Abflusswasser an den Zweigen und Schiften, welches bei langer dauerndem Regen nachtriglich diese Procente um einige Einheiten verbessert, ist wegen der Schwierigkeit der Untersuchung fir verschiedene Verhiltnisse noch zcht hinlinglich einwandfrei bestimmt. Die Vervielfiltigung der Oberfliche durch die Blatter namentlich aber die unzihligen Nadeln der Baume ist der raschen Verdunstung des aufgefangenen Wassers und damit dem definitiven Verluste desselben fiir den Boden ungemein giinstig. Von dem—nach Abzug von etwa 25-30% des Niederschlagswassers, welches an den Kronen der Waldbiume hiingen blieb, und weiteren 8-70%, welche durche Verdunstung aus dem Boden in die Luft verloren gehen,—iibrigen Quantum von etwa rund 60% werden ca 360 Dr. Hefele: 25% durch den Soten entgiltig aufgesogen, u. der Rest zur oberidischen Abfuhr gebracht, Jeder pflanzenbedekte Boden nun, ganz besonders aber der durch die Vegetation des Waldes in Anspruch genommene, erfahrt eine kolossale Verringerung seiner Feuchtigkeit, denn die Biume sind die grdssten Wasserconsumenten und darauf ist -eine Thatsache zuruckzufithren, welche gecignet ist, durch falschliche Auslegung eine gewisse Unruhe in waldkonservativ handelnden Kreisen hervorzurufen, ja vieleicht in der Hand gewissenloser Volksagitatoren und habgieriger Waldschlachter zu einem allerdings nicht einwandfreien Sturmmittel gegen die Erhaltung der Wéalder benutzt zu werden. Prof. Ototzkij in Petersburg fand, wie bemerkt, die wtbrigens aus kleineren Versuchen langst vermuthete Thatsache, dass der Grund- qwasserspiegel in der Ebene unter Waldungen eine bedeutende Senkung erfahre und somit praktisch das Gegentheil von der behaupteten Bodenfeuchtigkeitsbewahrung durch den Wald darthue. Sicherlich kann nicht erwartet werden, dass diese oft sehr bedeutende Differenz des Wasserstandes in tieferen Schichten zwischen Freiland und Wald zu Guusten einer reichlichen Speisung der Sickerwasser und damit der Quellen durch den Wald spricht. Bei gleicher geo-physikalischer Beschaffenheit leistet also die baumlose Adecxe mehr fir eine continuirliche Unterstiitzung des Grundwasserstandes und der Quellen als der Wald. Man ist fiir den ersten Lugenblick erstaunt itiber die drainirende Wirkung der Waldvegetation, aber cinige Zahlen mégen beweisen, wie kolossal der Wasserverbrauch durch die vegetativen Processe der Béume sich stellt. An sich ist die Produktion der organischen Substanz pro Jahr bei den Baumen schon grdsser als bei allen zdrigen Kulturgewichsen und daraus erklart sich auch der entsprechend héhere Wasserverbrauch durch Transpira- tion zur Erfiillung dieser gesteigerten Arbeitsleitung. Die im Baumkérper u. Blattern selbst aufgespeicherte Wassermenge ist ebenfalls sehr bedeutend, sie betragt nach Ebermayer bei einer kraftig entwickelten 85 jg. Fichte (im Holzkérper und in den Nadelen) ca t-ooo Liter; eine gleichalterige ‘Tanne hatte 1-200 Liter Wasser. Zur Produktion der organischen Substanz verbrauchte eine grosse Wald und Wasserwirtschaft. 361 Birke in 6 Monaten nicht weniger als 7080 kg. oder pro. Tag. 38 Liter im Verdunstungswege und eine 115 jg. Rothbuche beanspruchte ca 50 Liter in gleicher Zeit, wahrend bei jiingerem Alter (50--60 Jahre) eine Rothbuche pro. Tag. 10 Liter bendthigte. Ein Buchenhochwald producirt auf gutem Standorte jahrlich durchschnittlich 7057 kg. Trockensubstanz, was einer jahrlichen Wasserconsumption von etwa 2,187,670 kg. oder=218 m/m Wasserhohe gleichkommt. Diese Zahlen er6ffnen Einblick in den Haushalt der Natur von geradazu verbliiffender relativer Einfachheit und lassen die entsprechenden Schliisse auf den zzmeren Zusammenhang der iusserlichen Erscheinungen zz: Nimmermehr wird es gelingen einen schénen alten Buchenbestand zu erziehen, zo ihm das Minimum seiner zum Wachsthum néthigen Wassermasse nicht zu gute kommen kann. Es daszr¢ somit die ganze Abstufung der Bonitat bei gleicher physikalisch-chemischer Bodenbeschaffenheit fiir eine Species zu einem guten Theile auf den Grundwasser-bezw. Niederschlagsverhalt- nissen. Was nun fir die Ldene vom Einfluss der Waldes auf die Wasserverhialtnisse gilt, ist etneswegs zutreffend mit Zunahme der Erhebung des Bodens, also im Berglande und Gebirge ! Mag in den Ebenen immerhin der Wald verringert werden auf Grund dieser Beobachtungen, besondere Nachtheile gvossex Stils werden daraus nicht erwachsen, héchstens dass einige Quellen niederen Ursprunges versiegen. Grdssere Quellen verdanken ihre Entstchung und ihre Speisung meist dem Druckwasser der héheren Lagen und werden dadurch, dass trefere Platze entwaldet werden, kaum in ihrer Stirke und in dem hervorragenden Werthe, der ihnen fiir Wasserversorgung der Fliisse, Wiesenbewisserung etc zukommt, beintriichtigt werden. Mit zedem meter hoherer Bodenerhebung aber gestalten sich die Verhiltnissheinsichtlich der Bodenfeuchtigkeis im Walde giinstiger, cwdchst die Bedeutung, der Werth des Waldes. Nicht nur dte Niederschlige mehrer sich, sondern auch durch die Adnahme der Temperatur und damit der Verdunstung, sowie durch die gréssere Leckerheit des Waldbestandes die Swmme der der dem Boden verbletbenden Feuchtigkett im Wachsen be- griffenitst. Die Vegetationsdauer in den Hochlagen ist siirser und damit die Aunspriiche der Baumevegetation an das Bedenwasser geringer, die sinkende 362 Dr. Hefele: Durchschnittstemperatur und Zunahme der relativen Feuchtigke:t bewirken auch eine geringere Transpirationserésse. In ze héhere Lagen man daher in den bergen anstetgt, desto weniger wird die in der Ebene an sich dedeutende Differenz im Boden-Wassergehalt einer bewaldeten und unbewaldeten Flache, desto mehr entkraéften sich die Vorwiirfe gegen der Wald als einen Feuchtigkeitsverzehrer ohne Gleichen, desto mehr bekommt der Wald das Recht der Existenz und iberall, wo man gegen dieses Recht siindigte, waren die Folgen schreckliche. Die koustante Speisung der Bache & Flisse ohne excessive schiédliche Extreme wich stets zach der Entwaldung den abnormsten Gegensitzen und die Geschichte der Schweiz, Tirols, des siidlichen Frankreichs (Provence) Italiens, Griechenlands (und teilweise auch Japans) lehrt jedem Unbefan- genen die traurigen Folgen, welche die Vernichtung des dem Menschen von der Natur gegebenen schiitzenden Waldes nach sich zieht. Im Bergland und Gebirge besitzt der Wald eben nicht blos das Recht der Existenz, sondern er wird zur swingenden Notwendigket, da sich der Faktor des mechanischen Hindernisses gegen die oberfliichlich sum Abfluss kommenden Wasser hinsugesellt. Die Menge des in threm Oberflichenablaufe zerstorend wirkenden Niederschlagswassers zs¢ ex- perimentell auf ca 35% der Totalniederschlagsgrésse ermittelt und einer Vergrésserung oder Verminderung in sweifacher Weise unterworfen. Je absorptionsfihiger der Boden ist, auf dem der Oberflichenabfluss exfolgt desto, mehr wird sie verréngert und die Aufsaugungmenge erhoht, von der hinwiederum das fir die Quellenspeisung so wwéchtige Sickerwasser abhangt, welches im Durchschnitt 17-20% des Niederschlages beziffert. In positivem Sinne wirkt nun auf das oberflachlich zum Ablaut kommende Wasserquantum und zwar in gaz bedeutendem Masse die Mezgung des in Frage kommenden Terrains. Je weniger lang infolge der Schwerkraft das Wasser auf dem concreten Boden verweilt, desto weniger versickert, desto mehr nimmt die Oderflichenablaufwassermenge zu. Wo Wald fehlt, ist im geneigten Terrain, also im Bergland, ein Ansteigen der oberflichlich abfliessenden Wasserquanta auf 25-55% der Niederschlige, je nach dem Terrainwinkel, zu erwarten uud in vegetationslosen Gebieten der Gebirge gehéren 609§ und mehr keineswegs zu den Seltenheiten. Die a Wald und Wasserwirtschaft. 363 zerstorende Kraft bewegten Wassers wachst natiirlich mt seiner Menge und alles was dieser Thitigkeit einen Hemmschuh auferlegt, muss in conservirenden Sinne wirken. ; Hier greift nun der Wald wie keine andere Vegetationsdecke wirk- sam ein, theilt durch sein Wurzelsystem und seine Streudecke u. durch die hiebei implicite geschaffenen tausendfachen mechanischen Hindernisse die abfliessenden Wasserfaiden und zwingt sie durch Erschépfung zhrer Kraft zur Unschddlichkeit. Runsenbildung, Anrisse, Abschwemmung der Feinerde, Auswaschung der Verwitterungsschicht, kurz zede Art von Terrainangriff mit all den nachtheiligen Folgen werden durch sesn Vorhandensein vermicden und die Umwandlung von normalen Bdchen in Wildbache mit ihrer verderblichen Geschiebefihrung und den gefihrlichen Begleit— und Folgeerscheinungen verhindert. Unumstosslich ist durch die direkte Beobachtung und lange Erfahrung bewiesen, dass der Zerstérung des Bergwaldes die Vertrockuung, Verédung der Hinge und im Weiteren unter dem Einfluss der meteorischen Nieder- schlige die vapid zunchmende Abfuhr der Bodenkrumme bis zum nakten Felsen folgt. Damit entfallen in erster Linie die Szckerwassermengen fir die Qzellenspeisung und gleichen Schrittes mit der Deterioration des Terrains ist die Verstegung dteser fiir den Haushalt und die Bewdsserune von Ackergriinden etc so nithigen und wichtigen Feuchtigkeitsspender su konstatieren. Das fallt umso schwerer ins Gewicht als die oberflaichlich liegenden und durch kein so wiederschlagsreiches Einsickerterrain unter- stiitsten Quellen der tiefercn Lagen erfahrungsgemiss wahrend der trockenen Jahreszeit ihre Thiatigkeit erheblich verringern oder gar einstellen, wenn in den tieferen Lage etwa der Wald fehlen sollte. ~ Die unschiidlichen Bergbiche der dewaldeten Epoche erleiden eine successive Veréinderung tn-trockene Rinnsale welche nur zu Zeiten heftiger Regen Wasser fiihren, dann aber in gewaltiger Masse, da die zeitlich und raumlich verzégerte Zufuhr der Feuchtigkeit zu ihnen von den Hiangen nach dem Untergange des Waldes, proportional sum Verwilderungs und Ver- kahlungsprocess thres Einsugsterrains, impetuos und kurs dauernd wurde. Diese Umwandlung sv echten Wildbichen mit immenser Geschiede- fiihrung tritt naturgemiiss zuerst im Oberlaufe aller Fliisse auf, bleibt se 304 Dr. Hefele: lange dortsclbst bis su cinem gewissen Grade localisirt, bis der Hauptab- flusskanal im Gebirge mit all seinen Scitenbachen durch seine crodirende Thatigkeit, als Folge der grossen trreguléren Wasser und Geschicbefiihrung immer wefer und weter dic seinen Lauf begleitenden Einhange und Ufer in ahnlichem Masse dceznflusst, wie die feinsten Runsen es erstmals auf dem chemaligen Waldterrain thaten. Uferanbriiche, Hangetnstirsc, kurs weitgehende Veriwiistungen durch Untcrwaschung und Corrosion fihren immer wehkr Geschiebe und Fels_ trimmer dem Mittel- und Unterlaufe zu, bis endlich die Schutt und Triimmerwelle dic Ebene erreicht, und der an sich viel weniger schadlichen Uberschwemmung des Landes die Udcrlagerung mit Schuttmassen hAénzu- He esellt. Die Zeitdauer bis dicses Stadinm errcicht ist, bet dem die Aufmerksam- keit der Lewohkuer der Ebene auf dic unaufhaltsam progressive katastrophale Gestaltung der Wasserfithrung gelenkt wird, hangt natiirlich ab von der Grosse des Einzugsgcbietes und namentlich von dessen ¢cologischer l-ormation. Weiche Schicfer und Mer gclgebirge oder sandige Verwitteruugsschichten untermuscht mit groberen Gesteinstriimmen, geben nicht selten Veranlassung zum Niedergang einer “ J/whre.” Hier mengt sich das rasch abfliessende Nicderschlagswasser so stark mit dem leicht abschwemmbaren Detritus bei kurzen aber heftigen Gewitterregen (Wolkenbriichen), dass ein lava- artiger Brei statt cines Wasserlaufes zu Thale eilt. Dvese spesijisch schwercre Masse besitzt cin fotensirtes Angriffs-und Transportvermogen fiir /oses Gestin jeder Grosse vom hausgrossen Felsstiick bis zum kleinen Kiesel und die Verwiistungen des ihren Lauf einsiumenden Terrains sind entsprechend gesteigerte, abgeschen davon, dass kein Wasser tm Stande wére, solche Steinmengen nach Zahl und bes. Grésse 7x gletcher Zeit* zu Thale zu forden. Treffen solche clementare Gewalten mit ihrem Endeffekt in dcwohute Gegenden so ist die Verwistung von Kigenthumwerthen und die Gefahrdung von Menschenleben im ausgicbigsten Maase zu fiirchten. * Demontzey berichtet uns, von einer solchen Muhre, welche in eézem Gange mit 65000 cbm Wasser nicht weniger denn 169000 cbm, feste Masse zu ‘Thal brachte. Wald und Wasserwirtschaft. 305 Die Schweiz Tirol und Frankreich weisen genug verwiistete Plitze an den Miiudungen solcher Wildwasser auf, der Ortschaften sind nicht wenige, welche aus dem gleichen Grunde verlassen werden mussten ! In gemiassigt kiihleren Klimaten oder in den warmeren, wo die Elevation der Gebirge cine geniigend grosse ist, fallt die Verlangsamung der Schnecschimelse durch Waldbestocknng ebenfalls bedeutend ins Gewicht ; die wngehceuren Hochwasser aus naktem Terrain bei rascher Schneeschimelsze und die Gefahren der Lawinen unterstiitzen den aus dem vorigen wohl schon geniigend gerechtfertigten Auspruch auf Bewaldung des Ber glan tes. Ich denke, der Hinweis auf die Volgen der Entwaldung reicht hin, um das Mittel zu zeigen, wie all diesen Ubelstiinden begegnet werden kann, wenn auch die Zerst6rung Ieichter war als die Wiederherstellung. Man muss im jedem bergigen Lande, das eine gesunde und produktive Wasserwirtschaft im Interesse von Ackerbau Industrie und Landwirtschaft aufrecht erhalten will, eine entsprechende Behandlung und Bewirtschaftung des Waldes als fundamentale Forderung fir die Erreichung dieses Zicles verlangen. Wo der Wald im Berg u. Hiigelland auf seinem natiirlichen Standorte durch Eingriff des Menschen verschwand und verschlechtert wurde, muss die schleunigeste Wiederaufforstung in weitmbglichstem Umfange befiirwortet werden, wenu sich nachweisen lisst, dass Verschwinden von Wald und locale Stérungen der Wasserfithrung von Bachen und Fliissen im ursachlichen Zusammenhange stehen. Dabei sollte wohl gedacht werden, dass cine Bedeckung mit [le@/d schlechthin nicht geniigt, um die entsprechende Schutzwirkung auf das Terrain auszuiiben; sch/echte Waldbestockung hat eben auch nur ferdierse die giinstigen Wirkungen zu verzeichnen, welche dem gut geschlossenen und gepflegten Walde zukommen. Alle Manipulationen und Neben-Nutzungen, welche den Wald in seinem Gedeihen beintriichtigen, sind auch a/s e¢ne Beintrachtigung seiner Il trkung aufsufassen. Ubermissige und unverniinftige Weide hat in Frankreich wie in Italien, Tirol nnd der Schweiz den Bergwald herabgebracht und damit auch seine wohlthitige Kraft nicht zum kleinsten Theile ¢//userisch gemacht. Die Schutswaldecsetsgebung, welche cine Nutzung des Waldes 366 Dr. Hefele: beschrankt in solchen Lagen oder direkt verbietet und auf welche zs. B. auch in Japan mit solchen Stolze hingewiesen wird, hat ww einen Werth, wenn sie sich auch auf einen wzrkiichen Wald und nicht auf das Zerrdz/d eines solchen bezieht und notabene auch der Wel/e und die Organe vorhanden sind, um seine richtige Behandlung zu tiberwachen. Es ist nicht de Nutzung als solche in der Regel, welche Nachtheile zeitigt, sondern ade Art der Nutsung. Von diesem Standpunkt ist auch die Bewirtschaftung solcher Waldungen zu regeln. Kahlschlége sind namentlich in hohen und steilen Lagen unter allen Umstiainden zu vermeiden; ein regelloser Planterbetrich mit seiner schlecht verdeckten Ausschlachtung uad Verwahrlosung verdient eben so wenig den Namen einer Wirtsciaft wenn er gar noch wie hierzulande, oft unter dem Deckmantel ciner natiirlichen Verjiingung, besser gesagt Verwistung, segelt. Der Saumhieb mag auch hier wohl die besten Dienste leisten, wenn die Aufforstung zim auf dem Fusse folet. Wo die Wiederaufforstung nicht umgangen werden kann, wird man zu unterscheiden haben o—-—...- — 2 ———————<” 3 q ' E Wald und Wasserwirtschaft. 371 - » Astotov piv vVd@s” Beniitzte Literatur :—Weber, Anfgaben der Forstwirtschaft. Ney: Der Wald und die Quellen. Frauenholz: Bessere Beniitzung des Wassers und der Wasserliufe, Coaz: Lawinen. Koch: Das schnelle Anschwellen der Gebirgswasser, Ebermayer: Einfluss der Walder auf Bodenfeuchtigkcit etc ete. Wese: Uber die Wasserabenahme in den Quellen ete. Wang: Grundriss der Wildbachverbauung, a Bewegungsgesetze des Wassers, Demontzey : Wiederbewaldung der Gebirge, eG. Clic. Ueber Entstehung und Vertheilung des Kamphers im Kampherbaume. VON Homi Shirasawa, Ringakuhakushi. (mit Tafeln XXI—XXIIL.) I. Hinleitung. Der Kampher verdankt seinen Ursprung dem Sekret von Cinnamo- mum Camphora Nees. Diese Pflanze wichst in tropischen und subtro- pischen Lindern. In Japan kommt sie ungefahr bis 36° nord]. Breite vor, besonders aber an der Meereskiiste ; Shikoku, Kiushiu, Izu, Suruga und Kii bieten ihr den besten Boden. In diesen gegen Kialte gut geschiitzten- Gegenden, kommt sie neben anderen immergriinen Laubhélzern in ansehn- lichen Mengen vor. Leider gehen mit der immer zunehmenden Vermehrung des Kampherbedarfes diese Bestinde allmahlig zu Grunde; die grossen prichtigen Exemplare trifft man nur mehr in Tempelhainen. Manchesmal erreicht der Kampherbaum eine Hohe von 20 m. und einen Durchmesser vor iiber 2m. In Formosa kommt er in grossen Bestiinden in den Urwaldern vor; die Kamphergewinnung ist dort eine bedeutende Einnahmequelle der Regierung. Der Kampherbaum wiichst auch im déstlichen China in Tschi King und Kiangsi auf der Insel Chusan, siidlich von Shanghai, Itschung in der Provinz Hupe und auch der Insel Hainan. In diesen Gegenden kommt er in mehr oder weniger grossen Massen vor, wihrend Formosa als das produktivste Land in Kampher betrachtet wird. 374 Homi Shirasawa: Ueber die Ausfuhr von Kampher und Kampheroel von Japan vom Jahre 1888 bis 1go1 gibt uns das Kaiser]. Jap. statistische Bureau folgende Angabe: aus Japan ; aus Formosa ; Kampher, Kampheroel Kampher, Kampheroel Kin Kin 1888 4,555,469 —‘1, 149,299 1889 4,971,849 867,414 1890 4,463,881 778,902 ISQI 4,429,050 O2 1537 1892 3,064,005 699,836 1893 2,487,485 444,184 1894 2071876 427,251 1895 2,238,386 516,792 1896 1,617,660 558,858 4,395,921 5,701. 1897 2,608,242 1,094,910 3,174,206 65:75 1898 2,434,028 684,037 2,292,098 15,954- 1899 2,758,625 1,100,226 3,198,740 6,440. 1900 3,280,715 450,973 1,571,200 1901 4,165,757. 1,561,970 (1 Kin ist ungefahr 1 englisches Pfund.) Da von fast allen Lindern der Welt, Japan die Hauptkampherproduk- tion hat und der Kampherbaum hier so weit verbreitet ist, dirfte es fiir Forsttechniker und Botaniker speziell in wissenschaftlicher und prak- tischer Hinsicht von grésstem Interesse und von Wichtigkeit sein, ueber Herkunft, Entwickelungsweise und Vertheilung des Sekretes,, resp. des Kamphers etwas zu erfahren. Bei der Wichtigkeit des Kamphers fiir die Technik und die Weiterentwickelung seiner Produktion insbesonders, wiinsche ich mit diesem Gegenstande mich zu beschiftigen. Was nun die Behalter des Sekretes in den Pflanzenorganen anbelangt, so schrieb zuerst de Bary! in seiner ,, Vergleichenden Anatomie“: ,, Nach dem Bau sind die Sekretbehiilter zu unterscheiden: in schliuche d. h. aus’ 1 De Bary, Vergleichende Anatomie 5, 143,152 und 209, Ueber Entstehung and Vertheilung des Kamphers im Kampherbaume. 375 Zellen, welche ihre Wiande beibehalten, hervorgegangene, daher gewohnlich als Zellen bezeichnete und intercellulare, ihrer Gestalt nach entweder, Giinge oder Liicken, Hohlen zu nennende.“* Erstere unterscheidet er nach den Formen in ,,kurze und lange‘S und lIectztere in ,,schizogene* und »lysigene™ ,oder rhexigene’. A. Tschirch! welcher sich in den Ictzten zehn Jahren speciell mit den Sekreten der Pflanzen beschiftigt hat, nennt dic oelfiihrenden Zellen ,,Oelzellen‘? ; die schizogenen Liicken ,,Ocl-oder Harz- behalter“ (auch Oelraum) ; die lysigenen ,,Oel oder Harzlucken‘. Fiir dic schizogen entstandenen und spiter auf lysigene Art erweiterten Behalter hat er den Namen ,,Schizo-lysigene Raume* ecingefiihrt. ,,Oblitoschizogenc Ginge“ hat er sic genannt, wenn die Sezernicrungszellen der Oclbehilter erst nach Bildung des Belages obliterieren. Ueber diese Sekretbehilter, speciell dic Enstchungsorte des Sckretes, haben in letzter Zeit Tschrich? und dessen Schiiler cingchende Studien gemacht. Becheraz, ,,Ueber dic Sekretbildung in schizogenen Giangen", Bern 1893; Sieck, ,,Die schizogenen Sekretbehilter, vornchmlich tropischer Heilpflanzen“, Bern 1894; Lutz, ,,die oblito-schizogenen Sckretbehialter der Myrtaceen“, Bern 1895; Bierman, ,,Ueber Bau und Entwickelungs- geschichte der Oelzellen und die Oelbildung in ihnen*, 1808. Unter diesen Arbeiten hat Bierman im Speciellen ttber Oelzellen und die Oelbildung in ihnen bei den Pflanzen von den Familien der Lauraccen, (Cinnamomum Camphora ausgenommen) Canellaceen, Valerianaceen, Zin- giberaceen, Magnoliaceen, Myristiaceen, Piperaceen, Calycanthaceen, Um- belliferae, Convolvulaceen, Papilionaceen, Untersuchungen angestellt. In gar manchen chemischen und pharmazeutischen Werken finden wir Abhandlungen iiber den Kampher ; aber da die Quellen zu diesen meistens dieselben sind, beschriinken sie sich auf seine chemischen Eigenschaften und seine Anwendung, etc. Was aber die Entstehung und Vertheilung des Kamphers im Kampherbaum anbetrifft, so ist eine cingehende Untersuch- ung, besonders die mikroskopische Forschung, noch nicht bekannt. = Tschirch, Angewandte Pflanzen Anatomie 5, 460-530. 2 A. Meyer hat frither diesen Namen cingeftihrt ; Wissenschaftlich Drogenkunde 183, 5. 79. Tschirch, Harz und Harzbehiilter, 1900. Tschirch und Oesterle, Anatomischer Atlas der Pharmakognosic und nahrungsmittelkunde, 1909. 376 Homi Shirasawa: Flickiger! schreibt: ,,der Kampher findet sich auskrystallisiert in Spalten des Stammes, sowie aufgelést in dem Oel, welches in allen Theilen des Baumes (mit Ansnahme der Bliiten) verbreitet ist. Pfeffer? hat dic Ansicht, dass das aetherisches Oel der lebenden Zellen des Kampherbaumes * durch cine postmortale Sauerstoffaufnahme im Kampher verwandelt wird. In Weismer’s Buch steht? ; alle Theile des Baumes enthalten in besonderen Sekretzellen ein aetherisches Oel, aus welchem zum Theil schon in der lebenden Pflanze der Kampher im krystallinischen Zustande sich aus- scheidet. In Bezug auf den Gehalt von Kampher im Kampherbaum haben neuerdings Moriya? und Mimura durch Destiilation von Holz und Blattern werthvolle Ergebnisse, besonders mit Rucksicht auf Kamphergewinnung gehabt. Sie schreiben: ,,der Kamphergehalt im Kampherbaum ist sehr verschieden je nach dem Alter und Standorte des Baumes. Bei demselben Exemplar nimmt cr vom Wurzelstock nach den Aesten hin ab. Das Kernholz enthalt mehr Kampher als das Splintholz. Aus dem aus Formosa stammenden ungefahr 500 jaihrigen Holz haben sie 7.13 proz. Rohkampher gewonnen, wahrend sie aus dem 70 Jahrigen Holz, welches in der Provinz Bingo gewachsen ist, nur 4,73 proz. Rohkampher gewonnen haben. Die lufttrockenen Blatter enthalten durchschmitlich 3,75 proz. Rohkampher und 1,25 proz. IKkampheroel. Ueber den Kamphergehait in den Blaittern von Cinnamomum Cam- phora theilt uns M. Kelway Bamber*® im Royal Botanical Garden, Ceylon mit, dass er durch Destillation von lufttrockenen Blattern durchschnittlich 2,080 bis 2,425 proz. Kampher und 1,96 bis 1,05 proz. Kampheroel gewon- nen hat. In meiner vorlicgenden Arbeit wird es sich hauptsachlich darum handeln, nach mkroskopischer Untersuchung folgende Fragen zu beantworten. 1 Pharmacognosie des Pflanzenreiches, 1891, 5. 150. 2 Pflanzenphysiologie I, 2 Auf. S. 501. 8 Die Rohstoffe des Pflanzenreiches 1900. ‘ Journal of the chemical Seciety Tokyo, Vol. 21 No, 6 und Ringakushikwai-ZasshiZ1, 5 ‘The Indian Forester Vol, 28 No. 4, 1902 5. 163. Ueber Entstehung und Vertheilung des Kamphers im Kampherbaume. & NI NI 1. Aus welcher Substanz scheidet sich der Kampher im Kampherbaum aus ? 2. Wann macht sich das erste Produkt, aus dem sich der Kampher bildet, bei der wachsenden Pflanze bemerkbar ? 3. Inwelche Form geht das zuerst entstandene Produkt secundir iiber, und was sind seine mikroskopischen Eigenschaften ? 4. Wo wird das erste Produkt angelegt und wodurch charakterisiert es sich? 5. Wo vertheilt es sich in den Pflanzenorganen und in welcher Bezichung zu klimatischen und sonstigen Verhialtnissen steht seine Bildung ? 6. Kann man vom Standpunkte der Kamphergewinnung aus, den Kam- phergehalt im Baume mikroskopisch bestimmen ? Diese Untersuchungen wurden im Jahre 1900-1901 in der forstlichen Versuchsanstalt in Miinchen und hauptsachlich im Jahre 19q01-1902 im pharmazeutischen Institut der Universitat Bern und spiter im forstlichen Versuchsgarten in Meguro bei Tokio, ausgefihrt. Das lebende Versuchsmaterial stammte aus dem botanischen Garten in Miinchen, und Bern sowie auch aus dem forstlichen Versuchsgarten in Meguro bei Tokio; das trockene Material aus der Sammlung des Herrn Prof. Dr. A. Tschirch und aus meiner eigenen. Herr Prof. Dr. H. Mayr tiberreichte mir auch das trockene Material aus der Sammlung, die er auf Seiner Reise in Japan angelest hatte. Mein College Herr a. o. Prof. Mimura, Tokio hat mir verschiedenes Holzmaterial aus der Provinz Fukuoka besoret. Es ist mir eine angenehme Pflicht diesen Herrn, vor Allem Herrn Prof. Dr. A. Tschirch, unter dessen freundlicher Leitung ich hauptsichlich diese Untersuchungen im pharmazeutischen Institut der Universitat Bern ausgefithrt habe, dem inzwischen verstorbenen Herrn Prof. Dr. R. Hartig. und Herrn Prof. Dr. H. Mayr an der Universitat Miinchen, welche wahrend meines Aufenthaltes in Miinchen fiir diese Arbeit ihre giitige Hilfe mir gewahrten, meinen wiirmsten und verbindlichsten Dank auszusprechen. Tokio, September 1902. Oo “IJ o/8) Tfomi Shirasawa: , II. Entstehung. Knos pen. Um festzustcllen, wo sich die erste Anlage der Oelzellen vollzieht, und wo man ihre fritheste Entwickelungsgeschichte wahrnehmen kann, habe ich zuerst an den Knospen Untersuchungen ausgefihrt. Dazu diente mir frisches Material aus dem botanischen Garten Bern. Die diinnen wie auch die dickeren Schnitte in der Quer- und Liangsrichtung wurden direct in etwas verdiinnte 1 proc. Osmiumsiiure oder stark verdiinnte Alkannatinktur ge- taucht. In ersterer Lésung wurden die Schnitte nach einigen Minuten, in letzterer nach cinigen Stunden untersucht. Bei meinen mehrmaligen Unter- suchungen konnte ich die Oelzelle oder eine der letztern entsprechende Zelle niemals finden. Die flachkegelige Spitze wird vollstandig aus weichem meristematischen Gewebe gebildet. Schon direct hinter dem Vegeta- tionspunkte treten in unregelmiassiger Vertheilung einige Zellen auf, welche sich durch Grésse und rundliche Form von den ibrigen Zellen auszeichnen. Sie reagieren noch nicht mit Osmiumsaure oder Alkanma- tinktur, und der Zellinhalt ist etwas durchsichtig. Betrachtet man diese Zellen an den in Alkohol liegenden Priparaten so bemerkt man, dass ihre Zellraume mit einer farblosen homogenen Masse erfiillt sind. Diese Masse quillt durch vorsichtiges Zufliessenlassen von Wasser und schwindet in Alkohol. Mit Jodjodkali nimmt sie eine hellgelbe Farbung an. Dieses deutet auf Schleim.-- Wir wissen ja durch die Untersuchungen von Tschirch! und Biermann?, dass die Oelzellen als Schleimzellen angelegt werden. Unmittelbar hinter dem Vegetationspunkte, besonders an den Randpartien der Knospenachse finden sich die ganz differenzierten Oelzellen, welche durch Osmiumsiure oder Alkannatinktur nachgewiesen werden kénnen, Bic: 1 Unter denselben machen sich aber noch andere Zellen bemerkbar, 1 Die Harz und Harzbehiilter, 2 Ueber Bau und Entwickelungsgeschichite der Oelzellen, Ueber Entstehung und Vertheilung der Kamphers im Kampherbaume. 379 welche in den verschiedenen Stadien der Entwickelung der Oelzellen stehen. In das erste Entwickelungsstadiun, also in die Zeit des Uebergehens der Schleimzelle zur Oelzelle, fallt das Auftreten der ,,resinogenen Shicht* Tschirch’s,—diese Schicht stellt eine feinkérnige vacuolige Masse dar. In der Zelle ist die Wand mit einer Schleimmembran dicht belegt, wahrend die resinogene Schicht inwendig derselben aufliegt, Fig. 2, a. Die von den verschiedenen Partien des Schleimbeleges herausgehenden, bandfoérmigen Schlauche treten in der Mitte des Zellraumes zusammen. ito: Im Zusammenhang mit der successiven Entwickelung der resinogenen Schicht steht das allmahlige Verschmelzen des Schleimbeleges. Es ist jedoch auch moglich, dass diinne Schichten des Schleimbeleges oder Reste desselben zuriickbleiben. Das erste Auftreten von Oeltropfen konnte ich bald nach dem Ent- stehen der resinogenen Schicht feststellen. In letzterer fanden sich die Oeltropfen hiufig in der Mitte, jedoch auch in beliebigen anderen Punkten. Diese Oeltropfen wachsen bald zu ansehnlicher Grésse heran, was bewirkt, dass die betreffenden Stellen der resinogenen Schicht aufgeblasen werden und das Ganze in den Zellraum hineintritt. Die aufgeblasenen Partien zeigen manigfache Gestaltungen Fig. 3—7. Im weiteren Fortschritte der Oelbildung treten die genannten Partien miteinander in Contakt und erfitillen den ganzen Zellraum, lassen aber nicht selten Lumen in der Mitte odar an der Zellwand zuriick, Fig. 8—9. Diese Masse,—sammt dem in ihr befindlichen Oel—ist ziemlich fest. Bei Unter- suchung der Knospenachse bei trockenem Material konnte ich sie mit dem Messer zertheilen und ohne Formveriinderung aus der Zelle herausnehmen In einigen Fallen habe ich im Zellraum die resinogene Schicht stark aufgeblasen gesehen, welche dadurch ein beutelf6rmiges Ansehn erhielt, Fig. 10. Bringt man eine solche Bildung mit Alkohol in Beriihrung, so schrumptt sie bald zusammen. Ohne Behandlung mit einen Reagenz hat die obenerwahnte Masse ein hellgelbes, homogenes etwas lichtbrechendes oder schaumiges Aussehn. Osmiumsiiure fiirbt diese Masse dunkelbraun und kontrahiert etwas. Mit Alkannatinktur nimmt sie eine intensive rothe Fiirbung an. Durch die 350 Homi Shirasawa: Behandlung mit Alkohol. dehnt sie sich aus und es bleibt eine feinkornige Masse zuriick. .Zusatz von Wasser hat zunichst zur Folge, dass die Masse zu einem ovalen oder rundlichem Gebilde kontrahiert wird, welches ein triibes Aussehn besitzt. Diese Tatsache, welche den allgemeinen Gesetzen entgegengesetzt zu ‘ sein scheint, lasst sich vielleicht auf folgende Art erkliren. Bei Hinzufiigung von Alkohol tritt momentan Volumenvermehrung der schleimigen Masse durch die Lésung des festes Oels ein. Setzt man Wasser hinzu, so wird infolee Quellung die Oellésung herausgepresst, und tritt eine Zusam- menziehung derselben ein. Durch fortdauernde Einwirkung von Alkohol und Wasser wurde sie nach und nach klarer und zuletzt blieb eine schwammige, oft grobporige oder netzartige etwas lichtbrechende Masse zuriick. Diese Masse speichert sehr begierig Anilinfarben auf. So farbte sie sich mit Jodgriin-Eisessig leicht griin, auch nach Abwaschen mit Alkohol oder Glycerin wurde die Farbung ganz energisch zuriick gehalten, wahrend das andere Gewebe farblos blieb. Mit Jodgriin und Fuchsin firbte sie sich sch6n violet. In Chloralhydrat, conc. Schwefelsiure oder Salpetersiaure ist die Masse unlés- lich, Chlorzinkjod gab ihr eine gelbe Farbung. Diese Reactionen berechtigen mich, sie als ,,resinogene Schicht* zu betrachten, welche Tschirch als das Laboratorium der Oelbildung (Harz- bildung) bezeichnet hat. Was fir eine Rolle diese resinogene Schicht in der Oelbildung spielt, ist noch ein Ratsel, wir miissen heutzutag noch der Kraft des lebenden Protplasmas die Fahigkeit der Oelbildung zusprechen. Zur Zeit kann ich jedoch als sicher erkliren, dass der Ort der Sekretbildung bei dem Kampherbaum, wie bei den anderen Lauraceen in Tschirch’s resinogener Schicht zu suchen ist. Bei den Untersuchungen von Knospen Jhabe ich bei der ersten und zweiten Blattanlage, d. h. bei den innersten Schuppen nur Schleimzellen bemerkt. Auf dem Liingsschnitte sind sie meistens am iusseren Teile in verticalen Reihen augeordnet. In einigen Fallen aber waren bei der dritten Blattanlage die bereits ausgebildeten Oelzellen oder die Schleimzellen mit Ueber Entstehung und Vertheilung des Kamphers im Kampherbaume. 351 der resinogenen Schicht bemerkbar. Von hier nach den dusseren Blattan- lagen zu ist die Oelzellenbildung eine intensive. Was die Entwickelungsgeschichte der Oelbildung anbetrifft, so lisst sich ein wesentlicher Unterschied bei diesen zwei beschriebenen Fillen nicht erkennen. Blétter. Bei dem kaum entfalteten Blattchen von etwa 1!/,—2cm Gesammtliinge sind bereits die Schleim- und Oelzellen vollstindig ausgebildet und zwar in Blattspreite und Blattstiel. oO > Die Oelzellen sind mittelst Alkannatinktur oder Osmiumsiure nach- weisbar. Die Zahl der Oelzellen ist jedoch meistenteils noch verhiltniss- missig gering, wahrend sich vielmehr die Schleimzellen in grésserer Zahl bemerkbar machen. Bei den einjahrigen Blittern, welche sich bereits véllig entwickelt haben, weisen die Oelzellen schon ihre charakteristische Ausgestaltung und vollig entwickelte Grosse auf. Die verschiedenen Stadien der Oelbildung sind jedoch bei diesen nicht nachweisbar. Die Oelbildung in der resino- genen Schicht hat zur Folge, dass dieselbe stellenweise aufgeblasen wird und nach innen in den Zellraum eindringt und zwar von der Seite der Zellwand, der die resinogene Schicht entweder unmittelbar angelagert oder vor der sie durch eine Schleimmembranschicht getrennt ist. Diese blasen- artigen Gebilde sehen rundlich, eiférmige oder wurstf6rmig aus, Fig. 11, 12, 13, 14, 15, 16. Manchmal sind eine grosse Menge derselben beieinander angelegt und sie erfiillen dann den Zellraum, Fig. 17. Ohne Zusatz eines Reagenz haben sie ein fast farbloses oder hellgelbes, stark lichtbrechendes Aussehn. Durch Zufliessenlassen von Alkohol kontrahieren sie sich bald und bleiben hierbei Fetzen der resinogenen Schicht zuriick. Dieser Fetzen des resinogenen Hiutchens fiirbt sich mit ciner Liésung von Jodgriineisessig, aber die Farbung ist keine intensive, mit Jodjodkali nimmt er eine orangen- gelbe Farbung an. Unter den Sekretzellen dieser Blatter trifft man noch viel Oelzellen, welche auf ihrer Innenseite mit ciner dicken Schleimmembran nebst der ; 382 Homi Shirasawa : resignogenen Schicht versehen sind. Bisweilen erscheint die resinogene Schicht beutelf6rmig. Hiebei bliebt die farblose Membran durchsichtig und die resinogene Schicht sieht etwas trib aus. Mit Jodjodkali farbt sich die erstere hellgelb, wahrend die letztere eine orangegelbe Firbung annimmt, wodurch man das Vorhandensein der Schleimmembran und zugleich die Abgrenzung zwischen den beiden leicht wahrnehmen kann. Bei den zweijaihrigen Blattern sieht man an der Blattspreite ueberhaupt, und auch an dem einzelnen Geweben z, B. Epidermis, Zellwand, etc. eine bedeutend dickere und festere Struktur als bei den einjihrigen Blattern. Die Oelzellen sind bei zweiahrigen Blattern noch in den verschiedenen Stadien ihrer Entwickelung. Sie zeigen noch meistens Schleimmembran oder die Reste desselben, obwohl diinner und weniger zahlreich als in einjahrigen Blattern. Tropfenweise vorhandenes Oel habe ich nur in wenigen Fallen gefunden. Es bildet sich wie bei den einjahrigen Blattern in der schwammigen resinogenen Schicht, oder in blasenartigen Beutelchen. Ich konnte einen grossen Unterschied in der Entwickelung nicht bemerken ; ein Unterschied besteht jedoch darin, dass in den zweijaihrigen weniger Schleim, hingegen mehr Oel in der resinogenen Schicht vorhanden war als bei den einjahrigen. Ich bin daher der Ansicht, dass der Prozess der Oelbildung in den Blittern cin fortdauernder ist und zwar wahrend ihrer eanzen Lebensdauer, denn unter den zu meinen Untersuchungen beniitzten zweijahrigen Blattern zeigten sich stets die gleichen Entwickelungserschei- nungen, wiewohl einzelne der Blatter schon die auf baldiges Abfallen hindeutende charakteristische mattgelbe Farbung hatten. Meist war entsprechend dem Entwickelungsstadium der Blatter cin successiver Fortschritt in der Oelbildung zu bemerken. Bei den Blattstielen steht das Fortschreiten der Oelbildung fast immer im entsprechenden Verhaltniss zu den zugehorigen Blattern. Bei diesen habe ich wiederholt den tropfenformigen Oelinhalt in den Oéelzellen bemerkt, Fig. 18, und zwar ist derselbe in zweijahrigen Blattern reichlicher als in einjahrigen. In mehreren Fallen war aber die resinogene Masse mit Oel durchtriinkt vorhanden. o>) ve Lo) Ueher Entstehung und Vertheilung des Kamphers im Kamphorbaume. Durch Behandlung mit Alkohol lésten sich die Oeltrépfchen sehr bald, wahrend in den resinogenen Schliuchen sich das Oel erst unter dauernder Einwirkung von Alkohol léste. Schleimmembran oder die Reste derselben bleiben zuriick, jedoch fehlten sie auch manchmal, oder waren _ iiber- haupt diinner und geringer als in den Blattern. Die Blattstiele geben immer ein sehr zweckmiissiges Material fiir Untersuchung der Oelzellen, da wir bei denselben je nach Belieben einen diinnen oder dicken Schnitt bekommen kénnen und da auch wegen des geringen Vorhandenseins von gefarbtem Zellinhalte im Gewebe immer ein klares Bild wahrzunehmen ist. Bei der Untersuchung von Blattern und Blattstielen des trockenen Materiales aus Java stammend, habe ich auch Oel in tropfenformigem Zustande gesehen, Fig. 19. Auf Osmiumsaure reagiert es schwach, aber mit Alkannatinktur fairbt es sich sch6n rot. In Alkohl lést es sich bald, unter Zurticklassung von nur wenigem Riickstand. Reste der Schleimmembran waren bei diesen kaum bemerkbar. Auffallend ist, dass man oft in Knospenachsen, Blattstielen, Rinde, auch im Holz, netzformige oder hautartige resinogene Masse findet, welche fast die ganzen Zellriume ausfiillt. Bei Blattern fehlt dieselbe in der Oelzelle fast immer und ist bei frischem Material durch dicke Schleimmembran ersetzt. Fiir den Nachweis der Verkorkung der Oelzellwand, auf welche Zacharias! zuerst hingewiesen hat, habe ich Schwefelsiure, Chlorzinkjod oder Jodjodkali benutzt. Bei den jiingeren Pflanzenorganen wurde durch conc. Schwefelsiure das ganze Gewebe zerstért, wobei die feine Zell- wandlamelle der Oelzellen zuriickblieb. Bei den alteren Organen farbte sich die Zellwand durch Jodjodkali orangegelb. Rinde. Die von mir vorgenommenen Untersuchungen erstrecken sich aut frisches und trockenes Material, und zwar auf die jiingsten Teile einjahriger Triebe. 1 Botanische Zeitung 1879, 5. 617-645. Homi Shirasawa. wo IS Beziiglich der Oelbildung treten fast die gleichen Verhiltnisse hervor, wie bei den einjahrigen Blattern. Gelegentlich der Untersuchung der Rinde 5 jahriger Triebe war zu constatiren, dass der Prozess der Oelbildung schon zum Abschluss ge- kommen war. Hellgelb aussehende oft schaumige Oelmassen erfiillten das Zellinnere. Durch Behandlung der Oeltropfen mit Alkohol lésen sich dieselben unter Zuriicklassung einer farblosen schwammigen oder einer hautartigen Masse Hots. Ueber die Entwickelungsgeschichte der Oelzellen bezichungsweise die Oelbildung in demselben erfahrt man nur weniges, dagegen kann man bei demselben ausschliesslich die interessanten Umwandelungsformen des ent- standenes Oels zum krystallinischen Kampher studieren. Im Wasserpriparat des jungen 2, 3 oder 5 jahrigen Holzes sieht man nur den hellgelben Oelinhalt in der von den Nachbarzellen ausgezeichneten Oelzelle. Er erfullt den Zellraum oder einen Teil desselben. Mit Alkannatinktur farbt das Oel sich rot, durch Osmiumsiure wird es sebraunt. Alkohol lést das Oel leicht und bleiben manchmal membran- artige oder schwammige Klumpen zuriick. Beim ungefiihr 80 Jahre alten nnd vor 12 Jahren gefaillten Holz, welches aus einer bekannten Kampherproduktionsprovinz stammte, habe ich hauptsichlich die Bildung vom Kampher untersucht. Wenn man diinne Schnitte dieses Holzes im Wasser betrachtet, so sieht man vier verschiedene Formen: a. Dunkles oder orangegelbes Ocl b. hellgelbes Oel c. farbloses Oel d. - Krystalle. Das dunkle Oel ist cine balsamartige Masse; bei der mikros- kopischen Untersuchung tritt es oft aus der Zelle aus, wobei es seine > atk eh me A <-> whee ete eee a he ee —_————————— Ne Ee a ee Ueber Entstehung und Vertheilung des Kamphers im Kampherbaume. 385 urspriingliche Form beibehalt. Dasselbe hat meistens ein schaumiges, triibes Ansehn, Fig. 20 und erfillt fast immer den ganzen Zellraum. Alkoho! lést es langsam unter Bildung von zahlreichen feinen Hohlraumen. Durch andauerndes Zufliessenlassen von Alkohol nehmen diese an Zahl und Grosse zu, und bleibt zuletzt ein schwammiges netzartiges Skelett zuriick. Durch Aether oder Chloroform geht die Lésung schneller vor sich. Mit Alkannatinktur firbt es sich intensiv rot, mit Osmiumsiure braun. Das hellgelbe Oel ist dagegen etwas durchsichtiger und schliesst meistens in der Mitte oder in den der Zellwand anliegenden Partien grosse Luftblasen ein, Fig. 21. Alkohol lést es leichter als das vorige, bei der Lésung bleibt ebenfalls ein Riickstand. Das farblose Oel ist cine ganz durchsichtige homogene sehr fliichtige Flissigkeit. Es kommt in der Zelle tropfenweise vor, und habe ich nur in zwei Fallen ganz mit demselben erfillte Zellriume angetroffen. Es kommt bald in grossen bald in kleinen Tropfen vor, die in der Regel, der Zellwand anliegen, Fig. 27. Der Rand der Tropfen ist stark lichtbrechend. Oft treten in dem Tropfen kleine Luftblasen auf. Seine fliichtigen Eigenschaften kann man gut feststellen. Unter dem Mikroskop betrachtet verfliichtigt es sich schon in einigen Minuten, wobei ein in Alkohol léslicher kleiner Klumpen zuriick bleibt. Nicht selten findet man in der Mitte dieser Oeltropfen oder am Rande derselben kleine Krystalle, Fig. 28. Bringt man diese Krystalle oder die Tropfen selbst mit Wasser direct in Berithrung, so drehen sie sich sehr lebhaft und verschwinden schliesslich vollstindig. Dieses Oel firbt sich sehr schwach mit Alkannatinktur oder Osmiumsiiure. In Alkohol oder Aether ist es sehr leicht léslich. Die Krystalle treten 6fters in den Oelzellen des Holzparenchyms auf, wihrend in demselben Schnitte die Oelzellen der Markstrahlen und des Libriforms mit gelbem oder farblosem Oel erfiillt sind. Diese Krystalle sind farblos, weich und ist die Krystallform sehr undeutlich. Viele dieser Krystalle sind zu Aggregaten vereinigt, Fig 29, a. b. Dieselben lésen sich sehr leicht in Alkohol oder Aether ohne Riickstand. Durch Erwirmen verfliichtigen sie sich vollstiindig. (os) io) OV Homi Shirasawa: Die bei der Untersuchung angewendeten Methoden und Reaktionen sind folgende : 1. Kochen. Kleine Holzspane wurden in einem Becher zwei Stunden lang mit Wasser stark gekocht. Das farblose Oel und die Krystalle waren vollstandig vorfliichtigt, aber das gelbe Oel fast unverandert. Bei der Untersuchung von im eisernen Kessel mit Wasserdampf destillierten Holzspanen, habe ich dieselbe Erfahrung gemacht. 2. Erhitzen. Die Schnitte wurden auf dem Objecttrager einige Minuten lang langsam erhitzt. Hierbei verfliichtigte sich das Oel und die Krystallmasse gianzlich, wahrend der in Alkohol unlésliche mit Jodgriin und Fuchsin farbbare schwammige Klumpen zuriickblieb. ' 3. Sublimation Erhitzt man kleine Schnitte des Holzes im Sublimierglaschen schwach und langsam einige Stunden lang, so sublimiert die krystallinische Substanz (Kampher) und bildet feine Krystalle auf der kalteren Flache des Glaschens ; zugleich bilden sich dort farblose Oeltropfen, wihrend das gelbe Oel fast unverandert in der Zelle zuriickbleibt. Anstatt des Sublimierglaschens lassen sich auch ausgehdhlte Object- triiger verwenden, die Héhlung muss mit cinem Deckglaschen bedeckt werden, welches sorgfaltig anzudichten ist. Der Kampher krystallisiert auf der Innenseite des Deckglases, wo sich auch das fliichtige Oel niederschliagt. 4. ldrbungsreagentien. a. Alkannatinktur. Die in der sehr verdiinten fast alkoholfreien Lésung tiber ro Stunden Oo aaf eee Ueber Entstehung und Vertheilung des Kamphers im Kampherbaume. 397 lang liegenden Schnitte zeigen das gelbe Oel intensiv rot gefairbt. Die Krystalle und das farblose Oel sind durch das lange Liegen in der Tinktur nicht mehr sichtbar. b. Osmiumsiure. Die 1% Lésung derselben briunt des gelbe Ocl, dagegen wirkt sie kaum auf das farblose Oel ein. Jedenfalls ist die Einwirkung auf das in den jiingeren Organen enthaltene Oel stirker als auf dasjenige des alten Holzes. 5. Ldsungmittel. Die Léslichkeit des im alten Holz vorkommenden Oecels ist sehr ver- schieden. Im folgenden ist das Verhalten des Oels gegeniiber den einzelnen Lésungsmitteln wiedergegeben (Die Léslichkeit des Handelskamphers ist bei jedem Loésungsmittel angegeben). a. bey te) The Micrococcus present in the Eggs. Form. The cell cultivated in bouillon for 24 hours is 1 w in diameter. It occurs always in packet-form in nutritive fluids. But in the intestinal juice of the larvz it occurs in the form of diplococcus. Gram’s method: Positive. Oxygen: Aérobic. Bouillon: Bouillon becomes turbid little on the seventh day of inocula- tion, and a light yellow precipitate is formed after 20 day’s culture. Gelatine plate: Surface colony is yellow-moist, bright, elevated, round and sharply defined. By weak magnification it is granular. Deep colony is a yellow point. Gelatine is liquefied. Glatine streak: A yellow, clevated colony is formed along the in- oculated line, gelatine being quickly liquefied. Gelatine stab-culture: Colonies are formed discontinously along the inoculated line. Gelatine is liquefied at first in the shape of a nail, but after wards in the shape of a cylinder. ———E————— ee ee Investigations on Flacherie. 413 Agar plate: Surface colony is yellow, moist, bright, non-tenacious, elevated, round, sharply defined, and has a point on the centre. By weak magnification granular consistence is visible. Deep colony is a white point. Agar streak: An elevated, especially in the central line, moist, homo- geneous colony, the color of which is yellow shadowed with black, is formed. Potato: A yellow, moist, homogeneous colony is formed along the inoculated line on sixth day at 23°C. Milk: Milk is coagulated with much production of acid. Gas-production: Gas is not produced. fio "hi,o is not formed. Reduction: Nitrate is reduced to intrite. Acid: Acids produced in 14 day’s culture in glucose-bouillon at 20°C. was 0,33% calculated as lactic acid. Yellow pigment of the micrococcus is insoluble in water, alcohol or ether, but soluble in potash solution, which turns pale red by warming with addition of HCl. By these properties this microbe is recognized as Sarcina lutea Fliig-ge. Chapter Ill. The results of the experiments. Since in the intestinal juice of the diseased larva, an abundant growth of bacteria takes place, it is certain that this malady is caused by these microorganisms. But as these bacteria make luxuriant growth only in the intestinal juice and never invade considerably the tissues or blood, the pathogenic action will perhaps be due to the production of a certain toxin. Hence some experiments were undertaken to test this suggestion by using a solution, containing toxin, prepared in the usual manner from the culture d of the micrococci commonly found abundantly in the diseased larva. Experiment I. This experiment was performed in this College in October of tgo1. At 414 S, Sawamura. this time it was rather cold and since flacherie happens more rarely in cold than in warm weather, the silk-worms used for the experiment were reared in a large box constructed to keep the larva at a somewhat elevated tem- perature. This box and other apparatus used in the experiment were steri- lized with the vapors of formalin. The material used for this was prepared from Micrococcus LI cultured in bouillon for 9 days at 36°C. The filtrate was prepared from the above culture by filtering through Chamberland’s filter; and as in some cases toxin is not secreted from the living bacteria cells, a part of the culture was heated to 65°-70°C. for 30 minutes to kill the bacteria-cells. Oct. 29. 3 P.M. The original culture, the filtrate and the heated cul- ture were given together with mulberry-leaves to the larve of the second day of the fourth age. The number of the larve used for each experiment was 20, and the quantity of the materials used was 1,5 cc. to 100 ers. of mulberry-leaves. The larvae showed a very good appetite, and then they were treated as usual. The temperature in the box was 21°C. Oct. 30. When they were examined in the morning, there were no diseased larve found. Hence the culture, the filtrate and the heated liquid were given again to the larve as before, and the temperature was raised to 25°C. and water was besprinckled in order to increase moisture, because high temperature and moisture are favorable to the development of flacherie. But as the arrangement to keep the temperature high was imperfect, it fell too low during the night. Oct. 31. No symptom of the disease was observed in all the sections. Nov. 1. All the larvae, except one in the control experiment that had died, span healthy cocoons. An excreta of the larve fed with the culture of the micrococcus was put into bouillon, in which the micrococci made lux- uriant growth after a few days, proving that micrococci had entered and passed the digestive canal of the larva. Experiment 1. The negative result obtained in the former experiment might have been due to the low temperature. So this experiment was performed to repeat Investigations on Flacherie. 415 the former one using higher temperature. The culture used for this was also that of Micrococcus 7[ cultured in bouillon for 3 days at 36°C. Filtration and heating were performed as in the former experiment. Nov. 8. In the afternoon the materials were given twice respective- ly to 20 of the larvz of the fourth day of the fourth age in the same manner as in the former. The intestinal juice of the diseased larvze was also given to 10 larve. At 4 P.M. they were put ina thermostat and kept at 27°C. In the thermostat the ventilation was rather poor and the moisture content high, so that moulds grew on the excreta. The larve soon got into the stage of ecdycis, and on 1oth they ended ecdycis. From this day on death took place. Nov. 11. The silk-worms were transfered to a room of 21°C. The number of the dead larve will be seen from the following table. The heated | The intestinal Date. Control, The culture. | The filtrate. fee culture. juice. Nov. 10 I 4 I oO 7 EL oO ce) ce) Oo fe) Te fo) I fe) I fe) 13 fe) I fo) o 2 14 fe) I fe) fe) 15 fe) fe) fe) fe) I 16 fo) fo) fe) fe) re) 7 I fe) 2 oO fe) 18 fe) fe) fe) re) oO 19 fe) fe) fe) fa) re) 20 fe) fe) fe) re) re) 21 I 4 fe) fe) o Lc | a 3 fe) 4 I Ce) The remainder formed coccoons. As soon as the larve died, their intestinal juice was examined with a 416 S. Sawamura. microscope, and according to the microbes present in the juice and also other symptoms, the disease of the dead larve was grouped as follows :— Heated cul- Control. Culture. Filtrate. aa Intestinal juice, ure, Blacherie™ TE 4... ...0c: 6. S) i I — Seige (HL a9) Ben dae I (Grasserien es etersscore oO Pebring: Soi. poh eete 2 SE Gtaleee Qussceee 3 Flacherie in % of total TARVES 2 a2s2s ceaccuthecens 5 Contrary to the former experiment many flacherie-patients were pro- duced in this. It can, therefore, be concluded :— (1), that flacherie takes place when temperature and moisture are high and ventilation is insufficient, in short, when the conditions are injurious to the health of the silk-worms ; (II) that pathogenic action is not due to the production of toxin. Expertment ITT. This experiment was performed to confirm once more the result of the former ones. The cultures used in this experiment were prepared from Micrococcus IT cultivated in bouillon for 10 days at 36°C. and from JZicro- coccus J cultivated in bouillon for 34 days at 36°C. The filtrate was how- ever prepared only from the former. Nov. 14. At noon the cultures and other materials were given to the larve on the second day of the fifth age, taking 20 larve for each ex- periment. The temperature of the room was 15°C. and moisture 54. They were kept to the 18th, no diseased one being observed. They were therefore placed in a thermostat and kept at 27°C. and on the 22nd they 1 Jlacherie I denotes that in which micrococci were abundant, and flacherie If where bacilli were abundant, | Investigations on Flacherie. 417 were again transfered to the former room, and on the 26th they formed coccoons. The number of the dead larvz during the experiment was as follows :— Culture of Culture of ate. trol, : ; The filtrate, aes oe Micrococcus I, | Micrococcus II, are The disease was grouped as follows :— Control. Micrococcus I, \ Micrococcus Ll, Filtrate. iplgeberie: Ty .c.occces cases 3 5 2 7 =, © IDNR AgaKa GEG 2 4 I ° AGEOSSERIG PEE Aesth os fe) I ce) o IRE DEINE aes eeke incu esac 2 fe) o ° gS) ae 7 10 3 7 oT 7 of Flacherie in %% of the 25 45 1s ae COta le aRVEe Jc sec eesectes as ‘ a It will be seen from these tables that flacherie was more in the larve that were not infected artificially, than in those that received the bacteria. From this fact it can be learned that the bacteria, that cause flacherie, are already present in the vicinity ofthe larva and even in their intestines, wait- ing for an opportunity for development. Since many patients appeared among the larvz fed with the filtrate, flacherie would seem to be caused by 418 S. Sawammra. some toxins. But this can not be sure, because in the intestinal juice of the diseased larvae many micrococci were present which certainly had caused the malady. The results obtained in this and other experiments disprove the infec- tiousness of flacherie, which is against the belief held by -the sericulturists of the present day. Experiments IV. This experiment was performed to investigate once more the pathogeny of the micrococci. 1902. May 8. Jfecrococcus ff cultured in the decoction of mulberry- leaves for 7 days at 36°C. were given four times to 100 larve (Aohzki variety) of the first day of the first age. After this they were kept in the usual manner till the fourth age, without observing any symptoms of flacherie, The number of the larve examined on the first day of the fifth age were as follows :— Healthy Dead Lost Control 89 6 5 Inoculated on 3 6 The average temperature and moisture during the experiment were as follows :— Temperature Moisture First age 10,G°C. 68,4 Second. ;, elie 71,4 Third-'-,, 22,0 7593 Fourth ,, 20,0 78,1 Experiment V. Since flacherie occurs usually more in old larva than in young ones, the negative result obtained in the former experiments might be due to the fact Investigations on Flacherie. 419 that the bacteria were fed to young larve. Therefore this experiment was repeated, using old larve. The bacteria used for this experiment were Micrococcus [/ isolated this year from a diseased larva and the sarcina! isolated from the eggs of silk- worm. They were inoculated with the following materials. I. The micrococci, cultured on agar, suspended in water. II. The filtrate obtained from the decoction of mulberry-leaves cultur- ed for a week at 36°C. III. The above culture heated for 30 minutes to 65°C.? IV. The same to which formalin was added in the proportion of 1 drop tor xo cc. of the ‘culture.? May 22. 3 P.M. The materials above described were given together with mulberry-leaves+ each to 100 larva (Akahzki variety) on the first day of the third age. They were kept in the usual manner till the fifth age without observing any symptoms of the desease. The average temperature and moisture during the experiment were as follow :-— Date. Temperature. Moisture. May 22 22.50C 70,0 22 22; 78,0 24 2251 80,0 25 yA! 75,0 26 22,0 82,0 27 220) 69,0 28 18,5 7457 29 22,2 78,7 SS u Sarcina iutea Fliivoe ree 55 fe Nn Sterilized, 7 Sterilized, 4 1,5 cc. of the materials to 100 grs. of mulberry-leaves. 420 S. Sawamura. Experiment V1. As the result of the former experiments were all negative, it seemed doubtful that the bacteria used in these experiments were not the pathogenic ones. This experiment was therefore performed to observe the infective power of the intestinal juice of a diseased larva. May 28.3 P.M. The intestinal juice, obtained respectively from a dead larva whose body was softend and elongated, and from that whose body was contructed, were fed four times together with mulberry-leaves each to 10 larve (Akahzki variety) of the second day of the fifth age, In the intestinal juice there were of course bacilli and micrococci in great number. They were then fed in the usual manner for a week without observing any symptoms of the disease. The average temperature during the experiment was as follows :— Date. Temperature, May 28 17,07. 29 18,0 30 20,0 31 19,5 June 1 21,5 2 19,8 3 2059 From these experimental results it is clear that silk-worms do not become ill from flacherie when the surrounding conditions are favorable to their health, and they have resistance-power. These results agree with that reported by the Austrian Agricultural Experimental Station. Experiment VII. Since the negative results obtained in the former experiments might have been due to the insufficiency of the number of the bacteria, a further a Investigations on Flacherie. 421 experiment was made by injecting various bacteria directly into the intes- tines through the anus by means of a syringe, the point of which was care- fully rounded off. The bacteria cultured on agar were suspended in water and 0,05 cc. were injected. The species of the bacteria used were as follows :— Micrococcus J. ‘3 ALL, Coli-bacillus (isolated from the diceased larva). Bacillus mesentericus vulgatus Fliig- ge. » - Juscus r. Bacillus subtilis Cohn. June 7. 2 P.M. The above materials were injected into the larve (Aohzki-variety) on the second day of the fifth age. After the silk-worms had recovered the normal state which took about five hours, 10 lively larve were selected from each section, and at the same time 100 larve were kept as control, among which no disease appeared during the experiment. According to a previous experiment it was known that flacherie is produced after about three days even by injecting pure water into the in- testines through the anus, when the temperature is high, but when bacteria are injected the malady is produced more quickly. A slow development of the disease at a high temperature therefore would give naturally no decisive result. The results of the injection must be observed within 3 or 4 days. The results after three days were as follows :— 1. Distilled Water. The larve injected behaved very lively and showed a very good appetite. On the third day two of them died; in their intestinal juice many micrococci were found. 2. Micrococcus I. The silk-worn s lost appetite. On the afternoon of the second day four 422 S. Sawamura. of them died of flacherie ; in the intestinal juice micrococci and Lac. mega- therium were found in large number. On the same night one died, in whose intestinal juice only the micrococci were found. On the night of the third day two more died, in which also only the micrococci were found. 3. Micrococcus I. The larve lost appetite. On the afternoon of the second day one died of flacherie, in which much of Bac. megatherium and little of micrococci were found. On the second night three died, in which a great number of micrococci was found. 4. Micrococcus IT, The larve lost appetite. On that night two died of flacherie, in one of which the micrococci prevalent, while in the other Bac. megatherium exceeded the number of micrococci. On the second day five died of flache- rie, in which a great number of micrococci was found. 5. Colt-bacillus. The larve lost appetite completely. On the afternoon of the second day six died of flacherie in which coli-bacilli were found in great number. On the second night three died, and on the afternoon of third day one died. In the former only coli-bacilli, while in the latter Bac. megatherium was found. 6. Bacillus mesentericus vulgatus Fliig-ge. The larvee lost appetite completely. On the afternoon of the second day two died, in one of which Bac. mes. vulgatus prevailed, while in the other micrococci were more abundant. On the second night seven died, in six of which Lac. mes. vilgatus, but in one only micrococci were observed. 7 d Investigations on Flacheric. 423 7. Bacillus mesentericus fuscus Fliigge. The larvz lost appetite. On the second night seven died of flacherie, in four of which only Bac. mes. fuscus, but in three this microbe together with micrococci were found. On the afternoon of the third day one died, | in which Bac. mes. fuscus alone was abserved. 8. Bacillus subtilis Cohn. The larvze did not lose appetite so completely as the others. In the first night one died of flacherie, in which much Bac. subtilis was found. On the forenoon of the third day four, and on the afternoon one died of flacherie ; in the former Bac. subtilis prevailed, while in the latter micrococci. The results of the experiments may be summerized in the following table. First day. Second day. Third day, ies pele OU) MS | ne oe ee fe) fe) o = (EU ae fe) fe) 2 20 MIR OLOCELIS Elo Basho e'ctecde ness: fe) 5 2 es s LOE CR ee EMEC fe) 4 e) | 40 5 LA ry ee oe Peo 2 5 o 7° Coli-bacillus .........esseecesees 0 9 1 | 100 Bac. mes, vulgatus 0... 0.0... fe) 9 o | go to ee fe) 7 I So 03 (OE 7? (2 ee I fe) 5 60 From the results obtained in this experiment the following conclusions can be drawn :— 1. Many species of bacteria can propagate in the intestinal juice of the larve and cause flacherie. 424 S. Sawamiura. 1S) Disorder in the digestive organ such as injection of water causes flacherie. 3. From the above facts it is clear that bacteria, that’ can cause flache- rie, are present at all times in the intestinal canal of the larve wait- ing for an opportunity for development. 4. That flacherie caused by the injection of the bacteria is not due merely to the disorder in the digestive canal, is proved by the following facts. a. The bacteria injected into the intestines multiplied therein. b. When bacteria were injected, flacherie was produced more quickly and frequently than when water is injected. Experiment VIII. This experiment was performed to test once more for the production of toxin by injection. The materials used for the experiment were JZcro- coccus ITT cultured in a decoction of mulberry-leaves in absence of air for 7 days at 36°C. It was filtered through Chamberlana’s filter and a part of it was neu- tralized with Na, CO,. June 9. 2 P.M. 0,05 cc of the original and the neutralized filtrates were injected into the intestines of the larvae (Aofzki variety) on the fourth day of the fifth age. The results were as follows :— 1. Zhe Original Filtrate. Eleven larvae which received this material remained inactive for two hours. One of them was killed and the intestinal juice was examined, in which Bac. megatherium was found in large number. On the next morning eight larvae died, in two of which micrococci abounded, but little of Bac. megatherium was present; and in three others Bac. megatherium abounded, while micrococci were few; while in three others only Bac. megatherium was found. On the 11th two died, in which Bac. megatherium abounded, but few micrococci were found. Ww wi Investigations on Flacherie. 4 2. The Neutralized Filtrate. On the forenoon of the 1oth nine out of twelve larve used for the opera- tion died, in which Bac. megatherium abounded. On the 11th three died, in which both Bac. megatherium and micrococci were found. Experiment IX. Since some bacteria produce a powerful toxin only when they are mixedly infectcd, all the bacteria isolated from the diseased larvae were cul- tured together in nutritive glucose solution for 24 hours at 36°C. A culture of coli-bacillus also served. June 10. 11 P.M. o,1 cc. of,the original and neutralized filtrate of the above cultures were injected into the larve (Aohzki variety) on the fifth day of the fifth age. The number of the dead was as follows :— Number of or oF the the larve | First day. [Second day.| Third day. |Fourth day,| 70 = ate tested. | dead. Weiter atec ade hea k ces 2b 10 fe) fe) 2 fe) 20 Filtrate from _ coli- 7 bacillus teat cs. Bt 1p 9 P | 4 a ta The same neutralized. 10 fe) 4 | 4 2 Ico Filtrate from the mix- is é 6 ‘5 = @deculture: 2. ten: : The same neutralized, ie) fe) 5 2 2 90 According to the kinds of the bacteria found in the intestinal juice they may be grouped as follows :-— : ; rae Bac, megatherium Much Bae. Bac, megatherium | Bac, megatherium) 4 eoli-bacillus megathertunt, +miciococci, | +coli-bacillus, 4 micrococci. Sr | ee f VS ae o 2 o oO Filtrate from coli-bacillus, I 7 oO 2 The same neutralized ... fe) 3 2 5 Filtrate from the mixed : § 2 c Ls + - s The same neutralized ... 4 I fe) + 426 S. Sawamura. As many died of flacherie in this experiment, it might be supposed that the malady was caused by toxins, but that is very improbable, since water alone might have produced the same result.! Moreover flacheric is caused by various kinds of bacteria as was shown in the previous experiments. This makes it very improbable that a specific toxin is the cause of the malady. Experiment X. From the results obtained in the previous experiments there is no doubt that flacherie is not caused by a special toxin. But since the malady is caused by the multiplication of bacteria in the intestinal juice, the cause of the disease must be due to some action of bacteria and since many kinds of bacteria can produce this disease, the injurious action must be one common to all these bacteria. The vital action common to all of them and suspicious of injury to the silk-worm is the formation of acid, because the digestive enzyms of silk-worm are active only in an alkaline solution. The micrococci found in the diseas- ed larve and that in the eggs as well as Bac. megatherium, Bac. colt, Bac. subtilis, Bac. mes. vulgatus and fuscus, all produce acids in a solution con- taining carbohydrates, which were comfirmed by direct experiments. More- over, since the reaction of the intestinal juice is neutral or faintly alkaline, and the fluid excreted in flacherie is sometimes quite acid, there must exist some relation between flacherie and the formation of acids by the bacteria. Hence the effect of injection of acids was studied. 0,1 cc. of distilled water, 3% normal sodium carbonate solution, 29 acetic acid, 2% lactic acid and 2% butyric acid were respectively injected, as in the former experiments, into the intestinal canal of the larva of the fifth age. By this operation some vomited fluid, especially many ‘of those injected with water and sodium carbonate solution. All the larvae seemed somewhat inactive, but those injected with water and sodium carbonate solu- tion showed very good appetite after a few hours.? 1 Compare also the above experiment, p. 420. 2 The Jarvee injected with distilled water did not die within 24 hours, Investigations on Flacherie. 427 Those injected with the acids died with vomition and diarrhea after about 10 hours, the dead bodies softened, the third and fourth segments being elongated ; in short showing the close resemblance to those died of flacherie. Those injected with 0,1 cc. of 10% lactic acid died instantly without vomition or diarrhea, the bodies contracting and becoming rather hard. But even in this case the intestinal juice of the dead did not show an acid reaction, but still was alkaline, what shows that the silk-worm even dies when the alkaline reaction of the intestinal juice is a little weakened. By this experimental results it may be explained, why the appearance of the dead bodies of the larva in one case is different from that in the other. Vomition and diarrhea characteristic to flacherie is probably due to the fact, that as the intestinal juice is neutralized by the acids produced by the bacteria, the patient secretes more juice to restore the alkaline reac- tion on the one hand, while resorption is stopped on the other; hence the quantity of the fluid in the intestinal canal increases so much as to cause vomition and diarrhea. 1! Experiment XT. But the bacteria seem to produce a certain poison, although it is no toxin. It is a well known fact that the coli-bacillus reduces nitrate to ni- trite. But the production of nitrite in the decoction of mulberry-leaves by Bac. megatherium and the micrococci were also proved by the writer.? This experiment was performed therefore to observe the effect of nitrite on the silk-worm. July 4.9 A.M. 20 larve of the second day of the fifth stage were fed with mulberry-leaves moistened with a 109¢ solution of sodium nitrite for a day. Onthe next morning a larva died. 2 In higher animals also the secretion cf the intestinal juice is much accelerated by presence of acids, znge, Physiol. Chemie. 2 Mulberry-leaves contains often much nitrate, 3 1.5 cc of the solution to 100 ers, of the leaves. The larvze did not eat the leaves as usual, 428 S. Sawamura. They were then fed with the normal leaves, but on the sixth day two died with vomition and diarrhea, but bacteria were not observed in the in- testinal juice as in the case of flacherie. By injecting 0,1 cc. of 19§ solution of sodium nitrite in the usual manner, six out of seven larvze used for the experiment died instantly, the bodies of which were softened and stretched. Those to which only 0.05 cc. were injected, were, for 10 hours after the operation, in a somnolent condition. Then they became again active, but on the third day five died; the dead bodies becoming softened. With the intestinal juice of the larvz that died of flacherie, the usual nitrate reactions can sometimes distinctly be obtained. These facts make it clear that nitrite formed by bacteria is one of the in- jurious products that may contribute to the development of flacherie. Experiment XT, Since Bac. megatherium or Bac, coli are of general occurrence it is no wonder that they propagate also in the intestines of the silk-worms. But as to the micrococci it is different. As a micrococcus and Bac. megatherium exist in the interior of some eggs, they might come from the eggs as Pasteur and Macchiati supposed. But flacherie is usually prevalent after the fourth stage. Therefore it is very improbable that the micrococcus remains in the digestive canal for so long a time without developing the malady. More- over the micrococcus found in the eggs was quite different from those usually found in the diseased larve. 1902 June 23.1. Agar-plates were infected with small fragments of a mulberry-leaf. The colonies formed after two days were as follows :— The original plate: Colonies of a large bacillus (Bac. megatherium or Bac. subtilis ?) The second dilution: Numerous colonies of the large bacillus and micrococcl. 1 It rained two days before, Investigations on Flacherie. 429 The third dilution: Colonies of the large bacillus and white colonies of micrococcus. June 24. The former experiment was repeated, as on the previous day there had been a heavy rain. The results were as follows :-— The original plate: Colonies of the large bacilli and micrococci in- termingled. The second dilution: Colonies of the large bacilli and white and brown colonies of micrococcus. June 27. The experiment was repeated with mulberry-leaves of Hara- juku where silk-worms were never reared before. The results were as follows :— The original plate: Colonies of various bacteria covered the whole surface. The second dilution: Yellow and gray colonies of micrococci be- sides those of other bacteria. The third dilution: White and light brown colonies of micrococcus. To decide whether the micrococci of mulberry-leaves are the same as those of flacherie, it was necessary to observe their action on the silk-worm. July 7. 9 A.M. The micrococci isolated from mulberry-leaves and those of the diseased larvz were inoculated in the usual manner into the larvee of the fifth day of the fifth stage, and as they died, their intestinal juice was examined with a microscope. The results were as follows :— Number |... Second Third Total 96 of the of larvae, [First day.| gay, day. otal. dead. VETER? | OE henner oe ae 10 fe) I 2 | 5 3° Micrococcus TT of silk-worm ...... 10 fe) 5 2 7 7° Bae. megatherium from silk-worm, ae) 2 8 fe) | fe) 100 Micrococcus { from mulberry- i Vc Ee 10 ° 4 6 x0 aad Micrococcus LI from mulberry- * TRUE MERCER RR Sar c ck oésc ok ace = > 5 _ = at PPO VER. A cc'os sig ce en casey aks 36 ) 0 oO Gj < Micrococcus J from mulberry-leaves formed a white colony, while A/ferecocens ZZ a light brown colony on agar, 430 S. Sawamura. The dead larvze were grouped according to the species of the bacteria found in the intestines as follows :— Bac. megath- Bac. megath- re oe) ; ree , pee een erium and |24€- fnenalh mn anid Few few ony micrococci. | 6747 OM'Y. | coli-bacillus. acteria, NEMS Tee ae sonacae ae gcoscaac ce I I I fo) fo) Micrococcus IT of silk-worm, 2 3 I I fe) Bac, megathertum 6.00.10... oO fo) | 8 fe) 2 Micrococcus I from mul- rs 4 = 5 é berry leaves 2.) e = | | Micrococcus If from mul- ee eee 9 I fo) fo) fe) bery-leaves hae2:7=22-28-ss4-- From these results it follows that the micrococci present on the mul- berry-leaves can cause flacherie in just the same manner as those from the diseased silk-worms. It is therefore very probable that the micrococci found in the intestinal juice are the same as those found on mulberry-leaves. Experiment XIII. 1902 October. In order to observe whether the micrococci found in the diseased larve exist also on mulberry-leaves or not, micrococci were isolated from these leaves and their properties were examined. 1 ING: «41: Form: The diameter of the cell cultivated in bouillon for 24 hours is 1 x. Commonly two are united. The cells are colored by Gram’s method. Bouillon: At 15°-17°C. bouillon becomes turbid on the second day of inoculation, and on the seventh day a white ring is formed on the wall of the tube and a white precipitate is formed, the supernatant fluid becoming clear. After 20 days a feeble scum appears. Gelatine plate: Surface colony is white, round, sharply defined, lipped, 1 The leaves came partly from the College farm in Aomada, and yartly from a garden in Zokio where no silk-worms were kept for 30 years, a: - —oO Eee nyestigations on Flacherie. 431 moist and has porcelain-like lustre. A point of light brown color is in centre. By weak magnification the appearance is the same, showing a curled con- sistence. Deep colonies appear as white points. Gelatine streak: Colony light brown, folded, a film is found along the inoculated line, gelatine being liquefied. Gelatine stabculture: At 12°C. after 20 day’s culture, colonies are formed along the inoculated line, liquefying gelatine. Agar streak: White, homogeneous, moist, elevated, tenacious colony. Potato: At 23°C. elevated white colonies are found which on the sixth day turn brown and show granular consistence. Milk: At 30°C. milk is coagulated in 24 hours, acids being formed. Oxygen: Growth is better in presence of air. Gas: Gas is not evolved by cultivating in a nutritive solution contain- ing glucose for 7 days at 15°C. H2S: H2S is not observed in bouillon cultured for 24 hours. Reduction: Nitrate is reduced to nitrite. Acids: Azolithmin is turned red in pepton-water cultures containing 5% of glucose. This micrococcus is therefore probably AZcrococcus corona- tus Fliigge. No. 42: Form: The diameter of the cell cultured in bouillon for 24 hours is 0.8 p. Usually two are waited. The microbe is colored by Gram’s method. Bouillon: On the fifth day it becomes turbid, and onthe seventh day a yellowish brown precipitate is formed. After 20 days a feeble brown scum appears. . Gelatine plate: It did not grow on gelatine within 13 days at room- temperature (winter). Gelatine streak: Granules of white and light yellowish color are formed intermingled along the inoculated line. Their color changes afterwards respectively to yellow and deep brown. Gelatine is slowly liquefied. Gelatine stabculture: Thread-like growth to the bottom and liquefac- tion in the form of a nail. Agar plate: At 30°C. the surface colony is light brown moist, bright, 432 S. Sawamura. round, lipped, sharply defined, and the centre is somewhat elevated. By weak magnification its consistence seems to be homogeneous. Deep colonies appear as white points. Agar streak: White, moist, granular, non-tenacious colony which as- sumes after Potato a few days a yellow color. : On the second day a flat, dry, light brown, granular colony along the inoculated line. Milk: It is coagulated showing alkaline reaction. Oxygen: Aérobic. Gas: Hg Gas is not evolved. H,S is not formed. Indol reaction: Faint reaction. Reduction: Nitrite is formed from nitrate. ’ Acids: Acids are formed in glucose solution. This micrococcus is Aftcrococcus bicolor, Zimmermann, Form: Nos” 3 The cell cultured in bouillon for 24 hours has a diameter of 0.8 np. Two are usually united, but sometimes four, isolated cells are rare. It is colored by Gram’s method. J Zouillon: At 23°C. on the second day a little white precipitate is form- ed, the supernatant fluid becoming clear. It is the same after 20 days. Gelatine plate: Surface colony is dirty white, round, convex, with a white ring. becomes lighter towards the margin. Gelatine is slowly*liquefied. : By weak magnification, the centre seems deeply colored and it Gelatine streak: Hjsis nottormed: Reduction: Nitrate is reduced to nitrite. Acids: 0.013% of acid, culculated from ;the dissolved Ca O as lactic acid, is formed by cultivating in pepton-water containing glueose and Ca Co, for 14 days at 15-20°C. The yellow pigment is insoluble in water or alcohol, but soluble in potash solution. The color is not destroyed by HCl or H,50,. This micrococcus is the same as Wfcrococcus [11 of the silk-worm, and is probably Streptococcus bombycts of Macchiatt. No. 8. Form: The cell cultured in bouillon for 24 hours has a diameter of tz. Commonly two are united but sometimes four. It is colored by Gram’s method. Bouillon: At 15°C. on the fourth day it becomes a little turbid, and on the sixth day a precipitate settles. It remains the same after 20 days. Gelatine plate: Surface colony is white, round, elevated, sharply de- fined, and moist. By weak magnification it appears homogeneous. Deep colonies appear as white points. Gelatine streak: An elevated, white, mo‘st, homogeneous colony is formed. Gelatine is not liquefied. Gelatine stabculture: Thread-like growth to the bottom. Agar streak: A moist, bright dirty white, homogeneous colony along the inoculated line. — Nes aris alls ys Investigations on Flacherie. 437 Potato: An clevated, white, moist, bright, homogencous colony. On the central line it is more elevated. Milk: Milk is coagulated, acids being formed. Oxygen: Aérobic. Gas: Gas is not formed. Pi elsS, is formed. Reduction: Nitrate is reduced to nitrite. Acids: Acids are formed in glucose solution. This is the same as Micrococcus IT of the silk-worm, and is probably the Streptococcus Pastorianus of Krasstlschtschik. No. 10: Form: The cell cultivated in bouillon for 24 hours is somewhat oblong and 0.8 w. in the longer diameter. Usually two are united. It is coloured by Gram’s method. Bouillon: On the seventh day little white precipitate is formed, the fluid remaining clear. It is the same after 20 days. Gelatine plate: Surface colony is deep yellow, round, elevated, sharply defined and moist. By weak magnification it appears granular. Deep colonies appear as yellow points. Gelatine is not liquefied. Gelatine streak: Deep yellow, rather dry, homogeneous colony, which is much elevated on the central line. Gelatine is not liquefied. Gelatine stabculture: Thread-like growth to the bottom. Agar streak: It is the same as on gelatine, but the color is fainter. Potato: At 23°C. on the fourth day flat, light yellow, moist, homo- geneous colonies are formed along the inoculated line. Milk: Milk is coagulated, acids being formed. moxyeen: A€crobic. Gas: Gas is not evolved. Elest i f1.5 is formed. Reduction: Nitrate is reduced to nitrite. Indol reaction: A faint reaction. Acids: 0.01% of acid, culculated as lactic acid, was produced in pepton- water containing 5% of glucose and some Ca,CO, for 14 days at 15°20°C. 438 S. Sawamura. This micrococcus is JZicrococcus aurantiaca, Cohn. No.) 30: The colony of this micrococcus is dark purple. The properties are not yet examined minutely. Experiment XIV. Nov. 11. 1g02. On 10A.M. 0.05 cc. of water in which the micrococci above described, cultured on agar, were suspended, were injected into silk- worms of the fifth day of the fifth stage, as in the former experiments. As control distilled water was also injected. All the larve except those that received water, lost appetite and on the next morning excreted liquid feces. They were kept in the sitting room and after Nov. 19 they were placed near a stove. The number of the dead larve and the temperature during the experi- ment were as follows :— Number C 2 Pasi epuoeune of the 2/12} 6 bs 20! 21 | 22|22/24)\2 ‘Total. lead larvee, |22{12]13/14]15]1 \# 18] 19| 20] 21 | 22| 23 | 24).25 dead, morning | 14] 11| 11} 18] 16] 11} 9,5) 11 | 20] 13 | 18| 20| 20] 20 | 20 Temperature (c)1 evening | 22|—|—| 24] 20/13 | —|18|—]/18| 25] 20| 20] 20| 20 | i | | Gontroliee st sare- 15 © | 0} 9,10 |.0)}|/O") 0,0 On|"Os "Ol TOs KOR RON as are 13 Wiatelscacutves | 10.1.0] .0.| 0} \2 |'2 |:o | odio 1,o-/pa, pie ner en souleee 60 | STOR en Oy ae aeneree once 10 04) 0}, 04}.3 |, 0} 0 ]L0: [Orly hi} OF} Take Om osOnlan 70 | INO. 2 te hocuseesatess 10 OO: ] OO tes INOS ae ae gts 9 O | :0 |:0°) 1).4,) © | ake] 0'|,04] Oullo |) os ton Ros tones 100 ING; shies tes tarsnae 10 O/| 10 | |r 16) or |r. 10 01/0) Tao | hou tOnaoeheo 100 IN OR es eearccesses 10 OUT Sai) Tale ive 10 100 ING IG) Ss seanges damn aa 8 © | 504] 025/93, 1 aie | 04 |50.| 208 | hOn| amaer 8 100 NOP adae courses 10 ©) |.) tal Tae) |, |r kOe ones fe) 100 ING) Biden ba retanenes 10 On) O10) 0 fs Ou Or Oh eae ee 10 100 INO Ds se parr eRe 10 O10. | Osan s2a) 44) Os}s081 10" 2 Io | 100 Micrococcus from 10 Bales hice 1 | 10 100 the OQGS bicnesnes ; eile | 4 Temperature after the 19th was very low during the night, as the stove was not used at night, a Investigations on Flacherie. 439 The conditions of the larve in each section of the experiment were as follows :— Control. 15 silk-worms were kept for control. They were healthy and span cocoons on Nov. 25. During spinning two died of pebrine and flacherie. In the latter case many micrococci were found among green compressed frag- ments of mulberry-leaves. The average weight of the cocoons was 0.487 gr. Water. The larvz recovered after a few hours. Nov. 14. On the morning a larva died vomiting a yellow fluid, the body of which was shrunk. In the intestines large bacilli and diplococci were numerous. On the evening another died, fore-part of the body shrunk and back-part expanded. The fragments of mulberry-leaves in the intestines were green; here only large bacilli were found. Nov. 15. Onthe morning one died, body softened and stretched out. Large bacilli were numerous, and also some diplococci were found. On the evening another diced, body expanded in the middle part. The pieces of mulberry-leaves in the body were brown; streptococci were numerous. Nov. 20. On the morning one died, body softened, back-part black. No fragments of mulberry-leaves present ; some diplococci in the intestines. Nov. 21. Onthe morning one died, body softened and stretched out. Some slender bacilli were present. Nov. 25. The remainder (4) span coccoon, the average weight of which was 0.411 gr. No. T. After the operation the larve lost appetite considerably. Nov. 14. On the morning one died, the third and fourth segments _clongated, excretion of soft brown dung of a faint acid reaction. The frag- 440 S. Sawamura. ments of mulberry-leaves in the intestines were not compressed ; many large bacilli and some diplococci present. On the evening two died, one with shrunk fore-part and green compressed leaf-fragments, and bacilli and diplo- cocci; the other with brown leaf-fragments and numerous bacilli of various size. Nov. 19. On the evening one died, body softened and expanded in the middle part. Leaf-fragments not compressed ; diplococci numerous. Nov. 21. On the morning one died, body softened and expanded. The intestinal canal was filled only with liquid with numerous diplococci and few bacilli. Nov. 22. On the morning two died, one with swollen segments and black back-part. In the intestines few leaf-fragments with some diplococci. The other with softened blackened body and green leaf-fragments; diplo- cocci and bacilli present. Nov. 25. Three remaining larve span cocoons, the average weight of which was 0.390 er. By preparing a plate-culture from the intestinal juice of the diseased larva many white colonies of the micrococcus inoculated besides various others appeared. No. ae The larvee lost appetite after the operation. Nov. 13. On the morning five died. The first of them with some folds on the body; a few brown leaf fragments in the intestines and numerous diplococci. The second also with folds, brown lcaf-fragments and numerous diplo- cocci and some streptococci. The third also with folds, brown leaf frag- ments, numerous diplococci and some streptococci. The fourth resembled the third; streptococci were more numerous, while with the fifth diplococci again prevailed, all the other conditions being the same. Nov. 15. The remainder were unfortunately lost by an accident. By preparing an agar-plate from the intestinal juice of the dead larva many colonics of the micrococcus inoculated were formed. Investigations on Flacherie. 441 Now .3) Nov. 14. On the evening one died, body softened and stretched, faint brown leaf fragments and large bacilli and diplococci in great number in the intestines. Nov. 15. On larva died, body was faintly yellow, with some folds. Leaf fragments brown; diplococci numerous. On the evening three died, bodies softened and stretched. In the first the leaf fragments green, large bacilli and diplococci present. In the second brown leaf fragments, diplo- cocci numerous. In the third no leaf fragments but a few diplococci were found in the intestines. Nov. 16. Onthe morning one died, body stretched and leaf fragments green, diplococci numerous. All the other larve excreted liquid feces. Nov. 17. Onthe morning one died, fore-part of body somewhat trans- parent, back-part thin. The few leaf fragments green, few diplococci and pebrine-organisms present. Nov. 22. On the morning two died, both with softened body and many diplococci, the one with brown leaf fragments; the other with empty inte- stines, the back part of the latter larva was black. No. 4. The larvz showed poor appetite after the operation. Nov. 14. On the morning one died, body rather hard, leaf fragments brown and compressed, diplococci numerous. Nov. 15. Onthe morning three died, fore-part of bodies shrunk. Leaf fragments brown, numerous diplococci. On the evening three died, bodies softened, leaf fragments green, diplococci numerous. Nov. 17. On the morning one died, body softened, leaf fragments green, streptococci numerous. Nov. 21. On the morning one died, leaf fragments compressed ; no bacteria were observed. 442 S. Sawamura. Nov. 25. One died, body softened. Few fragments of leaves, many diplococci. By preparing an agar-plate from the dead larva numerous light brown colonies of diplococci were formed. Nia: = 15). The larvz lost all appetite by the operation. Nov. 12. At noon one died, excreting a brown fluid of a faint acid reaction from the anus. The intestinal canal was full of brown fragments of leaves ; numerous diplococci present. Nov. 13. On the morning two died, excreting a brown fluid of a faint- ly acid reaction from the anus. One contained many large bacilli and few diplococci; the other few small bacilli and micrococci. Nov. 14. On the morning one died vomiting a brown fluid, the third and fourth segments were elongated, the middle part of the body expanded, black lines appearing on the fourth and fifth segment, leaf fragments brown and compressed, numerous diplococci and some large bacilli present. Nov. 15. Onthe morning four died, bodies stretched and containing brown leaf fragments. In the first the back part of the body black and streptococci,numerous. Inthe three others streptococci were numerous. Nov. 16. On the morning two died, bodies softened. Leaf fragments brown, diplococci numerous. By preparing an agar-plate from the intestinal juice of the dead larva yellow colonies of diplococci were produced in large number. No:.. 6: The larvze lost completely appetite by the operation. Nov. 13. On the mording one excreted a light yellow fluid of a faintly acid reaction from the anus. Nov. 14. On the morning two died. One with stretched and softened body, and green leaf fragments. The second with shrunken body had ex- creted a yellow fluid of a faintly acid reaction from the anus, Leaf frag ments green, numerous diplococci present, Investigations en Flacherie. 443 Nov. 15. On the morning two died, bodies faintly yellow. In one of them leaf fragments were few and compressed ; diplococci and streptococci numerous. In the other leaf fragments brown, numerous streptococci and some large bacilli present. On the evening one died, the middle part ex- panded, leaf fragments brown and compressed, diplococci numerous. Nov. 16. On the morning one died, body was softened, leaf fragments green, diplococci numerous. Nov. 21. On the morning one died, body shrunk, diplococci present in large number. Nov. 22. On the morning one died, the fore-part shrunk, and back part blackened. Leaf fragments brown, numerous diplococci present. Wo: ! 927: Nov. 13. On the morning one died, body shrunk, leaf fragments brown and compressed. There were found a few micrococci. Nov. 14. On the morning one died, body softened, leaf fragments brown, many large bacilli and few diplococci present. Nov. 15. On the morning two died, one with softened body and few green leaf fragments, streptococci numerous. In the other leaf fragments were brown and compressed ; numerous micrococci and few streptococci present. Nov. 16. Three died, bodies expanded in the middle part. In the first leaf fragments green, large bacilli present. In the second the leaf fragments brown, streptococci and large bacilli numerous. In the third the leaf frag- ments brown, diplococci numerous. Nov. 17. On the morning one died, body softened and leaf fragments brown and compressed, diplococci numerous. Nov: 20. On the morning two died; one with softened and faintly purple-colored bodies, leaf fragments brown and compressed, bacilli nume- rous. In the other body softened, the back part blackened, leaf fragments brown. Some saccharomycetes and large bacilli were found. 444 S. Sawiamura. ING. 8: Nov. 15. Onthe morning one died, body expanded in the middle part, leaf fragments green, streptococci numerous. Nov. 16. On the morning three died. In one the middle part of the faintly yellow colored body expanded, leaf fragments brown with many diplococci. In the second body softened, leaf fragments green and com- pressed, short bacilli numerous. In the third body softened, leaf fragments green, numerous streptococci present. Nov. 20. On the morning two died, with softened bodies and brown leaf fragments ; diplococci numerous ; in one also bacilli present. Nov. 21. On the morning two died, one with contracted fore-part and a few diplococci; the other with the fore-part elongated and many diplocecci. Nov. 22. On the morning two died during the spinning of cocoon; bodies softened, the back part black, leaf fragments green. In one bacilli were numerous but diplococci few; in the other bacilli and diplococci were equally numerous. By preparing an agar-plate from the intestinal juice of the dead larva only white colonies of micrococci were formed. hh fo eae Nov. On the morning three died, two with the body shrunk, and one with the body clongated. One with many bacilli and few diplococci, and brown leaf fragments; the second with numerous diplococci, and green compressed leaf fragments; the third with brown leaf fragments, numerous diplococci and some large bacilli. Nov. 15. On the morning one died, body black, leaf fragments brown, diplococci numerous but streptococci few. On the evening one died, body softened, leaf fragments green, diplococci numerous. Nov. 16. Onthe morning four died, bodies, softened. With three of them small black spots appeared on the body; intestinal contents were ee ee eS ee Investigations on Flacherie. 445 green. In one of these diplococci, streptococci and some pebrine-organisms were observed. With the second large bacilli and diplococci. With the third streptococci. With the fourth green compressed leaf fragments, and streptococci numerous, large bacilli few. Nov. 20. On the morning one died, body softened with small black spots allover. Leaf fragments brown and compressed, diplococci numerous. By preparing an agar-plate from the intestinal juice of the dead larva, yellow colonies of diplococci were exclusively formed. Micrococcus from the eggs. Nov. 14. Onthe morning two died, in one the body stretched, the second and third segments yellow, and leaf fragments brown and compress- ed, few diplococci were observed. Inthe other the body was shrunk and hard, leaf fragments also brown and compressed ; sarcina was found exclu- sively. On the evening two died, leaf fragments brown and compressed, diplococci present. Nov. 15. Onthe morning five died, bodics softened with brown leaf fragments and numerous diplococci in every case. Nov. 16. One died, body softened, leaf fragments brown, few diplococci. By preparing an agar-plate from the intestinal juice of the dead larva, yellow colonies of sarcina were exclusively formed. From the results of these experiments the following conclusions were drawn. 1. The micrococci on the mulberry-leaves can cause flacherie, No. / being the least able to multiply in the intestines of the silk-worm. 2. Flacherie can be caused by this micrococcus, what can be proved by the fact that by preparing a plate-culture from the diseased larve the colonies of the inoculated micrococcus were formed in greater number or exclusively. 3. Since flacherie is caused by injecting water into the intestines, it is clear that the bacteria that can cause flacherie exist always in the intes- tinal canal, which fact proves also that the mulberry-leaves are the carriers of the germs. 446 S. Sawamura. wei he micrococcus in the eggs can cause flacherie. 5. The decrease of appetite of the silk-worm by the injection of water or bacteria can be observed also from the diminished weight of the cocoons formed. Average weight of a cocoon. Gontrtals. tr cactus Sere eos eee eee eer 0.487 gr. Water injected? 2) iat autarake teat eae nee O.411 Micrococcus Nao iwocwlared’ cs. eee 0.390 6. Any constant relation between the bacteria inoculated and the symptoms of the malady was not observed in these experiments. 7. When mulberry-leaves in the intestines are green, the reaction of the juice is alkaline, though weaker than in the healthy animal, while the brown color indicates that the reaction is neutral or faintly alkaline. 8. Bac. megatherium seems to multiply usually after the micrococcti had developed juxuriantly. 9. At low temperature the bacteria do not bring on flacherie very soon.!_ The cause will probably be due to the slow growth of the bacteria at the low temperature. General Conclusions. Irom the results of the series of the experiments above described the following conclusions were drawn. [. There is no doubt that flacherie is caused by the growth of bacteria in the intestinal juice. Il. The bacteria usually found in large number in the intestinal juice of the diseased larvae are various kinds of micrococci and two kinds of bacilli. In most cases micrococci only or together with few large bacilli are found. A short bacillus is also usually found along with the micrococci ; however cases in which the short bacillus alone is found are very rare. Besides these microbes there are many other kinds which exist in a small number in the diseased animals. 1 Compare Experiments VIL and NII, Investigations on Flacherie. 447 III. The large bacillus was identified with Bacillus megatherium, De Bary, and the short one with the coli-bacillus. IV. There exists in the interior of the eggs of the silk-worm usually a micrococcus and a large bacillus. The former was identified with Sarcina lutea, Fltigge and the latter with Baczllis megatherium, De Bary. V. Various kinds of micrococci usually adhere to the mulberry-leaves. The writer isolated from mulberry-leaves 10 species of micrococci. Nine of them were used for the experiments, by which it was decided that flache- rie is caused by these micrococci and the sarcina isolated from the eggs. The micrococci isolated from the dead larvz were identified with those iso- lated from mulberry-leaves. VI. Itis clear that the sources of the bacteria, which multiply in the intestines of the silk-worm and cause flacherie, are the mulberry-leaves ser- ving as food. But when the larve are healthy they resist the action of the bacteria. However when silk-worms are reared at high temperature or any disorders are produced in the digestive organs, the microbes multiply and cause the malady The greater number of micrococci in the intestinal juice is due to their abundance also on mulberry-leaves. VII. Flacherie is, as above explained, not caused by any special bac- teria, hence Macchiatis’ and Krassilschtschtk’s assumption can not be com- firmed. My observations agree with those of the Austrian Experiment Station that flacherie is not infectious. VIII. The true cause of the disease is the increase of certain products formed by the undue and rapid multiplication of various microbes. These products are in all probability no toxins, but they may consist of ammonia formed by protein decomposition, or of nitrite formed from nitrate con- tained in the leaves, or of acids produced from carbohydrates. Very prob- ably these noxious substances are sometimes acting together. I hope to settle this question satisfactorily by further investigations. 448 S. Sawamura. The author must express here his sincere thanks to Prof. Sasaki who kindly translated Italian articles for him, further to Prof. Loew and Prof. Kosat, and to Mr. Honda and Mr. Hayashi, Experts of Tokio Sericultural Institute who furnished him the larva and eggs of the silk-worms, and finally to Mr. Yamasaki, Assistant of the College. —_————> <0 <—____ Zur Physiologie des Bacillus pyocyaneus, Il, VON O. Loew und Y. Kozai. In Fortsetzung unserer friherer Versuche,' cine mdédglichst giinstige Nahrstoffloesung fir den Pac. pyocyaneus zu finden, in welcher trotz lecbhafter Vegetation keine Schleimbildung aber reichliche Enzymbildung statthabe, fanden wir folgende Loesung diesen Bedingungen entsprechend : Pepton. on A Glycerin. Clea: ss Magnesiumsulfat. 0:01. Dikaliumphosphat. oy ee Natriumbicarbonat. ou aie Chlornatrium. Ons 5, Das Magnesiumsulfat wurde sterilisirt bei der Infection zugesetzt. Wir vaviirten in dieser Loesung die einzelnen Bestandteile mchrfach und jedes- mal war das Resultat entweder cine langsamere Vegetation oder cine Verzogerung der Wiederauflcesung der Bacterienmassen.? In dieser Loesung lauft die Vegetation in 18—20 Tagen bei 25 —28°C. ab, wenn die Kolben nur sur Haélfte voll sind was behufs reichlicher Enzymproduction notig ist und jeden Tag kraftig ungeschiittelt wird, wobei unter Sauerstoft- absorption die gelbe Loesung tief griin wird. Est vom 13. Tage ab hort die Reduction des griinen Farbstoffs auf. Dic anfainglich reichlichen Bacterien- massen lésen sich bis auf cinen geringen Bodensatz allmalig wieder auf. 1 Siche diese Bulletins, Bd. 4, No. 4 und No, 5. 2 Wir erhéhten z, B, das Pepton auf 19%, das Dikaliumphosphat auf 0.4%, wir eliminirten das Natriumbicarbonat und setzten endlich die Chlornatriummenge auf 0.2 und 0.1% herab; auch versuchten wir dieses durch Natriumsulfat zu ersetzen, 450 ©. Loew und Y. Kozai. Diese zuerst von Enimerich und Law boebachtete Wiederaufloesung wurde von Conrad als cine Antolyse aufgefasst, was aber wohl nicht dem wirklichen Vorgange entspricht; denn Autolyse! ist die Gesammtheit der in einem Organ (oder Organismus) nach dem Tode stattfindenden fermentativen Vorginge, es wird also als characteristisch angesehen, dass irgend cine vorherige Secernirung von Ensym nicht stattfindet. Andernfalls ist der Vorgang eben lediglich eine gewohnliche Verdauung ; denn es ist doch z. B- ganz und gar irrelevant, ab ‘cin Magensecret den Magen verdaut, der es abgesondert hat, oder cinen anderen Magen. Bei der Wiederaufloesung der gewachsenen Bacterienmassen durch die secernirte Pyocyanase muss erst eine gewisse Anhiufung der letzteren in der Culturflissigkeit erreicht worden sein. Dann erst kann der Angriff auf die Nucleoproteide? der Bacterien- leiber Erfolg haben und wird dann auch das weitere Wachstum einge- schrankt und endlich ganz verhindert.3 Wenn aber die Bacillen auf festem Nahrboden (Glycerin-Agar) cultivirt werden, so bleibt jedenfalls das Enzym in den Zellen, wenigstens grossenteils. Dafiir spricht die Beobachtung von Krause,* dass der Presssaft des B. pyocyancus milzbrandheilend wirkt.* Die Bacillen waren auf Agarplatten cultivirt und die Vegetation nach 48 Stunden mit Platinspatel abgenommen worden. Nebenbei bemerkt muss dieser Presssaft kaum Toxin und auch nur wenig Pyocyanolysin enthalten haben; denn 3 cc. waren nach Arause cinem Kaninchen nicht schidlich. Bei dem Interesse, welches sich an die Pyocyanase knipft, suchten 1 Theobald Smith Vat bereits i. J. 1894 die verdauende Wirkung in sterilen Geweben von Thieren beobachtet; spiter haben Sa/kowsky, Facobi, Magnus-Levy, diese Erscheinung weiter verfolgt, Besonders interessant sind dic Resultate Conradi’s, 2 Krawkow (fefmeisters Beitrige 1, 530) hat Pyocyancus-Zellen mit verdiinnten Natron extrahirt. mit Essigsaéure dic Lozsung gefiillt und nach dem Reinigen das Necleoproteid analysirt, Er fand darin: C=52.73% ; H=6.91%; N=16.50%; P=2.11%; S=1.0%. In den Membranen) tard) ep C=46.2% ; Il=6.79§; N=88. Dieser Stickstoffgehalt deutet auf eine chitinartige Substanz, ‘Was auch von Lvimerling fiir die Membranen des Dac, fluorescens liquefaciens vermutet wird (Ber, Chem. Ges. 3.5, 702). ® Nach Svegwart (C, Bakt. 30, 573) werden die Nucleoproteide der Baeterien auch von Pepsin verdaut—aber erst nachdem die Bacillen ge‘étet sind, was jederfalls auffallerd ist, * Centrbl. f. Bakt. 57, No. 14, 5 ‘Typhus konnte beim Meerschweinchen damit nicht geheilt werden. tn — Zur Physiologie des Bacillus pyocyaneus, I, 4 wir nach ciner Mcthode, welche bei grosser Kinfachheit doch cin reineres Product licfert, als bisher méglich war. Unser Ziel ist noch nicht crreicht worden, doch mégen immerhin cinige Beobachtungen der Mitthcilung wert scin. Vor cinigen Jahren hat / Neuenberg) ber crfolgreiche Be- handlung der Staphylomykosis mit der Pyocyanase (Rohfermentloesung berichtet. Derselbe stellte, wic K. Vaerst? in seinen erfolgreichen Ver- suchen der Mitzbrandbchandlung mit Pyocyanase, diesclbe in ctwas verschiedenen Weise dar, wie Lmmerich und Loew, namlich durch Aus- salzen nach Erhitzen auf 58° (6 Stunden). Auf 1 L. der sechswoéchentlichen Bouilloncultur wurden 500 g. Ammonsulfat gegeben,? nach 24 Stunden das Ausgeschiedene ciner mehrtigigen Dialyse ttberlassen und dann dic Loesung im Vacuum zur Trockne gebracht. Dic alkalische Locsung wurde also nicht erst neutralisirt, und in der Tat haben uns vergleichende Versuche gezcigt, dass dieses vorzuzichen ist. Beim Versetzen mit Essigsaure* wird Kohlensiure frei, welche beim nachfolgenden Aussalzen in Blasen festge- halten wird, so dass eine sehr schaumige Masse erhalten wird, welche schweer weiter zu behandeln ist. Newenberg sowohl wie Vaerst verwendeten Bouillonculturen. Diese liefern aber cine sehr schleimige Flissigkcit,* welche beim Aussalzen auch den Schleim ausscheidet, der nun cinen grésseren oder gcringeren Theil der Pyocyanase mit sich reisst. Wird nun diese Ausscheidung der Dialyse unterworfen, um das Ammonsulfat zu entfernen, so bemerkt man cine auffallende Abnahme des Schleims, so dass man zur Vermutung kommt, es’ habe cin mit ausgeschicdenes Enzym (wegen nun grésserer Concentration) den Schleim durch Hydrolyse in nicht schleimige Producte verwandelt. Wenn die dialysirte Loesung dann im Vacuum cingedampft wird, so wirkt kein Schleim mehr st6rend, beim Loesen, resp. Injiciren des Products. Wir haben aus 1 Liter Bouilloncultur 1 Tabilitationsschrift, Bern 1900, 2 Centrbl. f. Bakt, 37, No. 7. 3 Eine miissige Vermehrung des Salzes bringt nur noch cine geringe Mehrausscheidung zu Wege. + Es ist nahe zu 1 promille Essigsiiure behufs Neutralisation notig. Diese Schleimbildung beruht vielleicht auf der Gegenwart milchsaurer Salze. Auch essigsaure Salze und Asparagin liefen schleimige Culturen, Pepton aber nicht, 452 0. Lew und Y. Kozai. nur 1.5 g. des ‘Rohferments erhalten. Prof. Nitta beobachtete, ‘nach Darreichung von o.1 g. desselben per os, bei einem Meerschweinchen keine Spur eine Temperaturerhéhung oder irgend welchen andern Effect, es waren also keine Substanzen vorhanden, die perv os hatten schadlich wirken k6énnen, was von cinigem Interesse sein mag, falls cinmal digses Rohferment zur Bekampfung von Bacillen (Cholera) im Darm zur Verwendung kommen sollte. Wir haben nun die Aussalzmethode auch bei Culturen angewandt, welche zicht schletmig werden, speciell bei der ecingangs erwahnten Cultur- loesung. Zehn Liter der 18 tagigen Cultur wurden zunachst mit Chloroform versetzt und cinen Tag stehen gelassen, um etwa noch vorhandene lebende Zellen abzutéten. Am folgenden Tage zeigte die Flissigkeit einen inten- siven Geruch nach Tsonitril, es musste also ein primares Amin in der Cultur gcbildet worden sein. Die klare Loesung wurde abgegossen, der letzte Theil filtrirt und in die Gesammtmenge der alkalisch reagirenden Fliissig- keit (10 L.) sechs Kilo Ammonsulfat ecingetragen und unter haufigen Umrihren bei 6—8° stehen gelassen. Es schied sich nach einiger Zeit eine flockige Masse an der Oberflache ab, welche abgenommen und durch Filtration und Pressen von der anhangenden Ammonsulfatloesung so gut wie médglich getrennt wurde. Durch dreitagige Dialyse wurde der Rest des Ammonsulfats entfernt. Schon beim Anrihren mit Wasser wurde bemerkt, dass sich ein grosser Teil nicht wieder léste, trotzdem wurde dic Gesammtmasse in den Dialysirschlauch! gegeben. Der unldsliche Theil war von ciner melaninartigen Substanz schwarz gefarbt, und enthielt neben blaugriinen Pyocyaneusfarbstoff noch cinen geringen Anteil héherer Fett- sauren, und etwas Proteinsubstanz. Das Filtrat wurde zuniachst auf verdauende Wirkung gepriift, aber nur cine ausserst schwache Wirkung auf gequollenes Blutfibrin beobachtet, selbst als noch 0.2% Soda zugesetzt wurde. Daraus durfte wohl der Schluss gezogen werden, dass die Pyocya- nase keine Albumosenatur besitzt,? sonst ware sie mit ausgesalzen worden. Bei den Versuchen von Newenberg und von Vaerst musste wohl die volumi- 1 Zu antiseptischem Zwecke wurde auch etwas Chloroform zugesetzt, 2 Wahrscheinlich tihnelt sie den Peptonen, ET ee a ee > _ a Zur Physiologie des Bacillus pyoeyaneus, IT. 453 (i. 2 hlcimmasse, dic ausgesalzen wurde, viel Enzym mit niedergerissen desshalb wohl der Schluss gerechtfertigt, dass bei nicht schlei- en dic Abdampfmethode (im Vacuum) der Aussalzmethode ist, da sic sicher dic Gesammtmenge des Enzyms licfert. Rens mT pmly oosdy Pa bi Savi | att Em iththsy ‘ 7 Hip reg! Fi rs) rare’ rio Py me “ f Uber den Kalkgehalt der Milchdriise, VON M. Toyonaga. Ich habe in meiner friiheren Arbeit iiber den Kalkgehalt der grauen und weissen Hirnsubstanz darauf hingewiesen, dass die Driisen im Verhialtnis zur Magnesia viel mehr Kalk enthalten als andere Gewebe des Tierkérpers, was jedenfalls mit der grésseren Zellkernmasse zusammenhinet. Es war in dieser Beziehung natiirlich von Interesse diese Untersuchungen fortzu- setzen, insbesondere weil in Bezug auf die verschiedenen Organe des Tierkorpers auffallend wenige Aschen-Analysen vorliegen, wiahrend in Bezug auf den PAanzenkorper diese ausserst zalreich sind. Ich habe zunachst die Milchdriise in Betracht gezogen, welche ins- besondere deshalb Beachtung verdient, weil ihr Secret in Bezug aut Mineralbestandtheile ganz ausserordentlich von dem Blute differiert ; so fand Lunge:— 100 Theile Asche enthalten : Tundemileh, Tundeblut. ig, © EOe7 Rel Na,O 6,1 45,6 Ca O 3444 0,9 Mg O 1,5 0,4 Fe,O, O,14 0,4 P, O; 3755 13.3 C] 12,4 35,6 Wir crsehen hieraus in Bezug auf den Kalkgchalt ganz cnorme Unter- schiede. In der Hundemilch berechnet sich das Verhiiltnis NeO; CaO= Fy 22.93 in Hundeblute MeO; CaO=)%, 2,25 456 M. Toyonaga. Ich beschriinkte mich bei der Analyse auf die Bestimmung des Kalks und der Magnesia, da besonders dieses Verhiltnis fiir die verschiedenen Gewebe sehr charakteristisch, und die Asche der-Milchdrtise iberhaupt noch nicht untersucht ist. Ich trennte bei der Milchdriise einer Kuh so gut als méglich das Bindegewebe von der eigentlichen Driisensubstanz ab und bestimmte zu- naichst den Wassergehalt, derselbe betrug 66.7%. Nun wurden 88,641 ¢ Trockensubstanz mit 5 g wasserfreiem Natrium- carbonat gemischt und verascht wobei das Weissbrennen wie gewodhnlich sehr lange dauerte. Die Masse wurde zunachst mit Wasser extrahiert und nach Entfernung des kohlensauren- und phosphorsauren Natrons der ausgewaschene Riickstand mit Salzsiure gelést, wobei cine Minimalmenge Kieselsiure ungeldst blicb hierauf die Lésung mit Ammoniak bis zu alkalischer Reaktion versetzt und dann mit Essigsaéure bis zu schwach saurer Reaktion vermischt. Hierbei bleibt ein geringer flockiger Niederschlag von phosphorsaurem Eisen ungelést. Aus dem Filtrat wurde nun der Kalk mit oxalsaurem Ammoniak gefallt und das eingeengte Filtrat vom Kalkniederschlag zur Magnesiabestimmung verwendet. Es wurde erhalten: Cas =O) F005 ie =0,2231 g CaO Me. JO; =Or5aa = =0,0566 ¢ MgO hieraus berechnet sich fur 1000 eile frischer Driise : 100 Teile der Trockensubstanz CaO =0,8401 Tcile 6.2507 eile MeO =0,2rs1 ”;, 0,0639 ,, = E ; ; : ; oo ic xo Se ? Vergleichen wir das sich hicraus ergebende Verhaltnis——— mit den fir Mg Milz und Niere von Aloy! gefundenen Zahlen und mit den Zahlen fiir das Muskelfleisch von Saugetieren, so ergiebt sich: Milchdriise, Milz, Pankreas, Niere. Siiugetier-Muskel, Ca ? Me 2497 6,79 4,05 1,84 0,34 allen Iss ist somit auch bei der Milchdriise wie bei der anderen Driisen der 4 Jahresbericht f. Thierchemie 30, 5, 492. Uber den Kalkgehalt der Milchdruse. 457 Calciumgchalt grésser als der Magnesiumgchalt, wahrend fir das Muskcel- ~ gewebe der Warmbliiter umgekehrt der Magnesium gchalt grésscr ist als der Calciumgehalt. Vergleichen wir noch die Mengen von Ca und Mg im Muskel mit denen in Milchdriise und Milz, so hat man ftir dic organische Trockensubstanz: Sdugethier Afuskel (Katz), Alidehdriise. Mi/z (Ribaut) Ca ———0.033% G73. % 0.141% Me— 0.109 ;, G.036';, 0.056 ,, Es ergibt sich somit, dass nicht nur der Calciumgchalt absolut groésser ist - in der Driise wie im Muskel, sondern auch dass der Magnesiumgchait dort LP weit geringer ist als hier. Ich werde meine Untersuchungen fortsetzen. ——_—_—~- ep -——____- -- Der Erntequotient, VON Oscar Loew. Inallen vollstandigen Ernteberichten aus der Praxis sowohl, wie den Ver- suchs-Stationen, wird ausser dem wesentlichen Erntebestandteil, wie Knollen, Wurzeln, Friichten auch noch dic Menge des Krautes oder Strohes angege- ben. Aus diesen Zahlen ersicht man abcr nicht sofort, ob sich das Ver- haltniss zwischen diesen Bestandteilen dem Mittel oder einem Optimum nahert. Behufs ciner sofortigen Beurtcilung dicses Verhialtnisses moéchte ich den Begriff des Erutequotienten cinzufihren vorschlagen. Er gestattet sofort zu erschen, ob unter den gegebenen Bedingungen (Boden, Dingung, Wetter, ctc.) cin mittleres oder optimales Verhaltniss erziclt wurde. Er gibt dic Hauptlcistung der Blatter, der wichtigsten Producenten organischer Materie, in vergleichbaren Zalen an, cr zeigt, ob dicse Organe ihre Aufgabe voll und ganz erfillt haben. Dieser Erntequotient a 100 S driickt die Ernte des wesentlichsten Bestandtcils k = Korner, Knollen, Wur- zeln, in Procenten der Blattsubstanz, des Stroh’s, s, aus. Man kann fir die Zwecke der Praxis dice Gewichte des lufttrocknen Krautes oder Strohs zu Grunde legen, wahrend fiir rein wissenschaftliche Zwecke das Gewicht der absoluten Trockensubstanz zu dicnen hatte. Is wire von cinigem Vorteil, den absoluten Erntewerten pro ha auch den Erntequotienten, der sich bei normalen Pflanzen oft zwischen genau bestimmten Granzen bewegt, beizu- figen. So betraigt derselbe im Mittel bei Gerste 73, wahrend er im Optimum, wie es wohl in der Praxis nicht erreicht wird, too betragen kann.! Bei Bohnen liegt er in der Regel weit itiber too, bei Erbsen hiiufig-iiber 200. 1 Jieliriege? teilt mit, dass er unter sehr giinstigen Bedingungen im Glashaus bei Gerste gleiche Gewichte Stroh und Korner geerntet habe, Andererscits beschreibt Z. Wolf Feldversuche, welche auf 100 Stroh nur 60 Thi. KGrner gaben und bei Weizen gar nur go, 460 Oscar Loew. Ferner wiirde cr. sich im Mittel aus zalreichen Daten ergeben fiir Weizen Bena eR yuna et nh ee 53 AIOE: ee ONG eg eas cea rack ae een ee 66 IVES 2 So Ge ee rac aera te: wien er rae ae sa ina, OO Seti: Re ee ee lee ces eects ea 52 Buch weizen:. ©. i 2 cee eee ea eet oo eee eee 54 Wohl haben schon verschiedene Forscher den Kornerertrag hie und da auf 100 Theile Stroh bezogen, aber es ist systematisch weder der mittlere noch der optimale ,, Hrutequoticnt “ bestimmt worden. Besonders war es P. Wagner, welcher seine Resultate mit Cerealien in dieser Form ausdriickte. So fand er z. B. dass bei verschieden starker Stickstoffdiingung auf 1oo Thl. Stroh resultiren kénnen bei Hafer 51-87 Thl. Korner, bei Roggen 46-53, bei Weizen 33-63. Ferner hat er bei Hafer auf 100 Thl. Stroh 52 Thi. Kor- ner erhalten, als er Chilesalpeter bei der Einsaat gab, aber 64 Thl. Korner, wenn er diesen bei beginnendem Schossen zuftigte. ! Gewisse Verhialtnisse fithren zu einem Uebermass von Blattproduction, andere wieder erméglichen den Blittern, dic von ihnen bereiteten organi- schen Niahrstoffe in ausgicbigster Weise der Ausbildung der Friichte zu- kommen zu lassen. Diese Arbeit mit einer Zal auszudriicken, beabsichtigt der Ernutequotient. 1 Die Stickstoffdiingung der landwirtschaftlichen Culturpflanzen, 1892, 5. 164. — 2 JInauguraldissertation, Erlangen 1897. EE ———— Ueber die physiologische Wirkung des Chlorrabidinms auf Phanerogamen. 403 Versuch mit Brassica chinensis. Drei Tépfe mit je 1 Kg. Boden wurden gediingt mit: 1 g. Kaliumnitrat, 0.5 g¢. Ammonsulfat, und 0.5 g. Monokaliumphosphat. Ausserdem erhielt Topf a, 10 Milligramm Rubidiumchlorid. pene, Sor 83, ~ 5 ¢. diente zur Controlle. Der Chlorgehalt des Bodens entsprach nahe zu 0.05 g. Na Cl per Kilo, er war ein lehmiger Boden, zum Teil aus vulkanischer Asche bestchend. Am 21, October wurden 10 Samen pro Topf ausgesit und am 5. No- vember die jungen Pflanzen auf je drei méglichst gleich grosse, 6-7 cm. hohe, reducirt. Gegen Mitte November ergab sich, mit Ausnahme einer Pflanze in b. fiir die Rubidiumpflanzen ein besseres Wachstum als fiir die Control- pflanzen, ein Unterschied, der mit der weiteren Entwicklung immer bedeu- tender wurde. Am 17. December ergaben die Messungen fiir das lingste Blatt jeder Pflanze Folgendes : — | b. | c. Controlpflanzen. 21.0 cm. L420 | 16.5 ZQA- 55 FGI | 17.0 Poros 8 25.5 | A Am 22. Dec. wurden die Pflanzen ausgezogen, die Wurzeln gercinigt und mit Fliesspapier gut abgetrocknet, und die ganzcn Pflanzen gewogen im frischen Zustande, mit foleendem Ergebniss :— a b. vet WP ae Ae 6.1 10.1 L7,, 14.0 10.2 18.8 ,, 25.2 15.0 Mittel: 16.6 Reet 11.8 464 Oscar Loew. Es war somit ein stimulirender Effect des Rubidiumchlorids zweifellos, doch war dieser bei Erhdhung von 1o Milligramm auf 50 pro Kg. Boden nicht vermehrt worden, die Pflanzenmasse war im Gegenteil in Jetztrem Falle etwas kleiner als im ersteren. Versuch mit Gerste. Zwei Topfe mit je 1 Kg. lufttrocknem Boden erhielten ais Grunddiing- ung je 1.5 g. Ammoniumsulfat, 0.5 g. Monokaliumphosphat, 1.5 g. Calcium- superphosphat, 1.0 g. Kaliumcarbonat! und 0.5 g. Natriumnitrat. Einer erhielt ausserdem noch o.2 g. Rubidiumchlorid, der andere die aequivalente Menge Natriumchlorid. In jeden Topf wurden am 14 October 10 vorher gequollene Samen ausgesit und die Entwicklung im Glashause wie beim vorigen Ver- such beobachted. Am 21. October wurden die Pflanzen auf 4 pro Topf re- ducirt, so dass alle von méglichst der gleichen Héhe waren. Gegen Ende November zeigte sich cin deutlicher Héhen-Unterschied zu Gunsten der Rubidiumpflanzen, der stets zunahm. Die Messung am 17. December ergab :— enn EE ya ne Une Rb- Pflanzen. | Control- Pflanzen. 44.6 cm. | 39.1 cm. 46:95 HOO, AOS’. 5; 47-0 55 54-3» 49-5 » Die Héhen-Unterschiede nahmen zu, wie die am 19. Januar anfgenom- mene photographie (Tafel XXV) gut erkennen lisst. Dabei waren die Rubi- diumpflanzen vollstiindig normal.? Wegen Auftretens von Pilzen wurden die Pflanzen schon bald nach der Bliitenperiode geschnitten. Das Gewicht betrug : 1 Pas Kaliumearbonat wurde spiiter separat dem Boden einverleibt | 2 Ob Buchweizen und andere Pflanzen hiebei ebenfalls normal bleiben, soll noch gepriift werden. Ueber die physiologische Wirkung des Chiorrubidiums auf Phancrogamen., = 405 Rubidiumpflanzen, Controlpflanzen. eminent, WE SISCHOCWICHE 52.11. 0.2. ens noes Poe ie +e asa eee cy as Meeeetee Blather, {SCH 2 iec) sco ee fps ense dene es ORAS ye Bryn eh 6 hy ae mpeestorbene Blatter, lufttrocken; ......:..... Ne Uae AD 3 Ein dritter Versuch wurde mit Sfzxacea oleracea angestcllt. Alle Ver- haltnisse waren hier die gleichen wie oben bei Brassica. Bei den Rubidium- pflanzen erhielt der Boden 50 Milligramm Rubidiumchlorid per Kilo. Als der Samen reif war, wurde geschnitten und die besten Exemplare frisch ro) RKubidiumpflanzen., Controlpflanzen, Pe wieme cet erossten Fflanze, Varietat/l ...... 18.2 ¢.............. 12.0 g. Die zwei gréssten Pflanzen der Varietat Il. ... 16.5 g....... hee BRL2 Es hatte somit in allen diesen Fallen cin stimulirender Effect des Rubi- diumchlorids stattgefunden, was wohl von betriachtlichem theoretischen In- teresse ist. [tir die Zwecke der Praxis jedoch ist eine Anwendung des Salzes ausgeschlossen, da dessen Preis ein zu hoher ist.! 1 ks kosten roo g. Rb Cl=1z2 Mark (ca. 6 Yen). =—— »~oo —-—-— On the Stimulating Action of Manganese upon Rice. BY M. Nagaoka. In our last Bulletin the observation was communicated that small doses of manganese administered as sulphate had a very favorable action on the development of various plants. This made it very desirable to carry on a field experiment with rice which is the most important agricultural plant in Japan. Thirty six wooden frames cach representing an area of 0.826 square Meter were placed, three fect apart, into the paddy field of our College farm to a depth of 60 cm., leaving 6 cm. above the ground. The soil had not re- ceived any manure the previous three years’ and was now manured! in the ratio of 100 Kg. N_ per ha, as ammonium sulphate Pere ic. ,, 4, as potasstum carbonate ee ee. 5. 9,, ‘a8 double superphosphate. The potassium carbonate was separately applied (June 23) and the other two salts four days later. On June 29 manganese was applied as manganosulphate in such quanti- ties that the amount of manganic oxid corresponded to the following propor- tions, three series being observed in cach case :— 1 Before manuring the soil was sifted, and all remnants of former vegetation removed. 468 M. Nagaoka, OL Mn, O, per ha, Mn, O, per frame, No of wooden frames. Kg. Gram, ees or a | z Rees 1 13 | 25 re) o 2 14 26 fo) | fe) 5 15 27 10 0.833 4 16 28 15 | 1.250 5 17 29 20 1.666 6 1S 30 | 25 2.083 7 19 51 30 2.499 8 20 a2 35 2.916 | 9 21 33 | ite) 3:332 10 22: 34 15 3-749 1] 23 35 So 4.165 12 24 36 55 4.582 NSS SS a ee On July 7 the young rice plants (55 days old) from the scedbed, were transplanted into the frames, each receiving 16 bundles of twelve healthy individuals of equal size.1 The treatment (irrigaton, etc.) did not differ, from that usually observed with the rice fields in Japan. The weather con- ditions were not favorable this year for this crop in the whole Empire of Japan, but the relatively low summer temperature diminished on the other hand the dangers from fungi and insect pests with this crop. Our frames re- mained free from such pests. The crop was harvested on November 29 with the following result, obtained by weighing in the air day condition. 1 The variety was the Sa/swma, characterized by its resistance power and medium duration of vegetation, _—_- On the Stimulating Action of Manganese upon Rice. 409 Mecot Mn, O, | Full Empty | Straw. | AVERAGE. ‘oa per ha. | grains, | grains, | oe | Babe i on kg. | gr | oe gr. grains. | grains, ci 1. rot I Ke manure | 151.6 Be 193.0 13 | and no | 142.6 3.0 | 171.0 | 150.3 3-0 1850 338.3 25 | Mas Os) ) (556.5 27 | O10. | | | | | | | 2 He Mi Oste 177-0.. | 4.6 ley 924270 | ar | 227.8 | 6.3 | 312.5 | 20> 5a 5.4 269.6 477.5 26 | | 202.6 | Gee | 254.0 | | | | | | 3 | 250.6 6.8 319.6 | Bena to. ||) °23016 5.8 285.6 247.3 7.0 | 3087 564.0 27 | 251.8 | 8.4 321.0 4 249.9 6.3 401.6 16 15 | 257.6 | 4.8 BO7-5 256.7 4.4 329.1 590.2 28 | 262.6 | 5.1 278.0 | | 5 | ecto ia bs3 354.6 | yen eae, ©2604 | 78 341.6 | 264.3 6.3 327.7 598.3 29 | | 245.6 | 5.) 287.0 = 6 279.0 | 10.3 330.6 18 25 | 264.5 5.1 335.0 | 272.1 6.8 348.5 627-4 30 | 272.7 | ne | 326.0 | | | | 7 | 270.0 i ay 19 | 3° | 256.5 | 4.4 316.0 | 267.7 4.2 340.9 612.8 31 | 276.8 5:3 | 332.0: | aera | [treads a8 Wie 8 | | 264.6 | 8.2 325.8 20 35 | 2678 | 4.4 | 334.0 267.3 6.0 322.9 596.2 32 | 269.4 | 555 | 309.0 ( 470 M. Nagaoka. * 7 : Sb ae AVERAGE, Na ae Mn, O, Full Empty Straw. per ha. orains. grains, Full Empty frames. ke. or, er, er, outa eS Straw Total. : : : grains, grains. 9 261.8 8.0 364.6 21 40 269.3 5.6 308.0 272.3 6.9 338-5 617.7 33 285.9 7.0 343.0 10 256.5 9.0 340.6 22 45 286.5 7.1 338.0 271.9 7.1 334-9 | 613.9 34 272.8 5.3 326.0 s 2 | : II ? (198.6) (gio) |) are5 | | as 50 270.6 6.7 399.0 278.1 6.6 | 35o:5 (| 2 Gag 35 287.6 6.5 367.0 | 12 254.5 6 331.4 24 55 279.4 5.2 364.1 272.6 4.1 345.2 621.9 36 283.9 6.4 340.0 | a The application of manganese had therefore a considerable influence upon the yield, which will be noticed more conveniently by the following table, in which we take the yield in grains of the manured plot without manganese as a unit: Mn,O,, Kg. per ha. Ifarvest of full grains. (Average) 1109) 9 (=e ee OIE Onceomae Por cpor se (Cri soe. 1.00 DS) caine cate ww mates times v0s.sfor stabi cy as he Magis ore eee aa 1.20 LD” oe tat ka mes ore re wep aren 2) 014 lee ere Kuo « e\babinisck) 4 oor ceiekata tage ocean 1.30 BG! Ys apeustak POS aeyRd aes ae isda ree ee 1.34 20) ciuvead awe gem uimnidwelsies QisiWrimin'es sx o6iee'e axe shed faretn Seine [32 Se lh sain stance Saal Cecio n rcs Fares hae Tae {ae LO vencccccreererser reeves eareorenccerereectsisserueres 1.34 ND ccceureecnceseteeseneetreessers creer tne Meneateeenenss [.34 SO seccccrerereveerensencensearecseeteeeceseerceseusanees 1.37 On the Stimulating Action of Manganese upon Rice. 471 It will be noticed from these figures, that a» moderate dose of 25 Kilo Mn,O, per ha led to an increase of the harvest of one third and that higher doses of Mn,O, did not influence essentially this result under the eiven conditions. It is further of some interest to examine whether the average ratio between the weight of grain and straw is affected to any extent by the in- fluence of manganese. The quotient of yield! reece which expresses the percentage of grain relatively to straw is for the different cases :— MANURED PLOTS. Amount of Mn,¢ y, per ha. Quotient of Yield. INOM Mc SAUCESE 3 ceeseaccessges Meets soceetee te. 75 10 Ke. So \ j 1S 7> | 20) 5, 82 | 25 79 => iol! yy 1] 3 35 S2 2: I 40 So nr 45s S} SORT bs5 77 | Bey as 79 ; The application of manganese had therefore—ceter?s partbus—a favor- able influence on the quotient of yicld. Let us now determine by calculation whether the application of mangano- sulfate would be profitable for the farmer. The price of 100 Kilo of pure crystallized manganosulphate is according to the latest pricelist of Theodor Schuchardt= 110 Mark or 53 yen.? The average production per ha, of grains of rice with husk is= 3525 Avlo and of airdry straw = 5250 Kilo. 1 On the Quotient of Yield (Erntequotient) see the article of O, Zeezw in this Bulletin. ( ] 2 1 Mark =38,38; sen, latest quotation. 472 M. Nagaoka. The wholesale price of,crude rice grains is 9,9 sen per Kg., of airdry straw=1.2 sen, per Kg. hence the average yield per ha. has a value of 349 yen in grains and 63 yen in straw=412 yen. An increase of one third would have an additional value =137.33 yen while the cost of the mangano-sulphate required would be=30 yen. Hence the application of this salt on soils poor in manganese would be of advantage. The impure manganous chlorid of commerce would fulfill the same purpose and would cost less than 10 yen in the above case. On the Physiological Action of lodine and Fluorine Compounds on Agricultural Plants. BY S. Suzuki and K. Aso. A. On the Influence of Potassium Lodid on Oats. By S. Susuki. I have demonstrated in a former article’, that potassium iodid in exceedingly high dilution can exert a stimulant action on plant growth. The fea had served for that experiment. I had, however, at the same time commenced an experiment with oats, the result of which are described in the following lines. Soil and manure were exactly the same as in the former case: each pot contained 2300g. air dry soil and was manured with 3g. Na NO,, 3g-K,CO, and 4. 6g. common superphosphate. The seeds were sown (15 in each pot) on Feb. 21 and the young shoots reduced on March 7, to five per pot of equal height. Pot No. I. received on March 11 and 25, April 14, 21 and 28 and May 6, each time 0.01g. potassium iodid dissolved in 100 c.c. water ; further pot No. II. o.oo1g. and No. III. o.ooor1g. of that salt, while No. IV. served as control. Those quantities of potassium iodid expressed in percentage of soil are: Nea 0. Lo - = O60 2606" 9 ING, : IT. ==) 0.600, 2609 .9¢ No. III. = 0.00002609 % Bull. College of Agricultare, Toyko Vol. 5, No. 2. p. 199. 474 S. Suzuki and K. Aso. In the beginning of May, the tips of the leaves of No. I. turned reddish yellow and further growth was retarded, but an increase of shoots made up for the loss in height, leading finally to an increase in the yield compared with the control plants (compare the photograph, Plate XXVI). The plants were irrigated almost daily with 300c.c water until the flowering stage was reached, after that with 500c.c. The flowering period was over on May 16. The plants were cut on July 6. The straw and the grains, unhusked, were weighed in the air dry state with the following result : ah Sanne Ue II. II. IV. Number of Stalks. 15 14 14 9 Weight of grains, un- r3 prea, Mes husked, g. Ee Blo, 21-2 ah Weight of straw, g. 48.5 56.6 58.4 45.2 The result undoubtedly proves a stimulant action of iodine, even if present in such a small quantity as 2. 6g KJ in 10000 Kilogram of soil as in No. III. The increase however, is with oats not so large as with the pea (These Bulletins, vol. V p. 199). I had mentioned already in my former article, the experiment of A. Velcker who soaked seeds of wheat and barley for a short time in a 1% solution of sodium iodid and observed with such seeds, an increase of yield. The quantity of sodium iodid that penetrated into those seeds must then have been exceedingly minute, otherwise a poisonous effect would have shown itself. I have repeated that experiment with oats. The seeds were soaked for 24 hours in a 1% potassium iodid solution, washed and then sown in two pots, 15 seeds in each. Later on the young shoots were reduced to five of equal height. After a few weeks, it became clear that the plants did not so well develop as the control plants. This may be due to more iodid having entered into the grains than in the case described by Veeleker. This difference is probably caused by the prolonged soaking in my case, Also differences of temperature during the soaking process can influence the result. The plants were cut on July 13, and the straw and grains, unhusked, weighed in the air dry state with the following result : On the Physiological Action of Iodine and Fluorine Compounds on Agric, Plants. 475 I II Control, Weight of grains, unhusked, 18.1 g. | 16.8 g. | 21.4 g. } | Weight of straw, 24.4 5, 22-5: 35 | AS 2s, This shows that the amount of KI absorbed in the soaking was large enough as to cause a retarding influence, which was much greater, however, in regard to the production of straw than in that of grains. A field experiment further was made with oats. On 3 plots, each measuring 20 square meters, an equal amount of oats grains previously soaked for two days in water was sown on March 21. On April 15, the young plants had reached 3—4 cm. and were treated now the first time with potassium iodid solution!. The treatment was repeated on Apr. 22, May 7 and 22, and June to. The total quantity of potassium iodid applied to the plot No. I. was 0.25¢., to the plot No. I. 0.025g. On June 26, flowering commenced, on July 16, some spots of rust became visible. On August 6, the plants were cut, but owing to several storms, some loss of grains had occurred ; hence the final weight is somewhat below the actual production. The straw and grains, unhusked, were weighed in the air dry state with the following result : L IT. Control. Weight of straw and grains. 6.96 Kg. 6.08 Kg. 6.05 Kg. Weight of grains. ©1605; DSi css 0.83 A small increase of yield had therefore taken place by the application of 0.25 g. KI for 20 © Meters, while 0.025¢ had no influence. 1 The solution was highly diluted, each dose of potassium iodid being dissolved in to titres of water, 476 S. Suzuki and K. Aso. B. On the influence of potassium todid on radish By S. Suzuki. The same plots! which had served for the culture of oats just mentioned served for this experiment with radish. One plot received 0.5 g. potassium iodid in one dose that is double the quantity of that of the last ex- periment wish oats, the next plot received 0.05 g. potassium iodid in one dose (also double of the last experiment). The radish seeds were sown Oct. 1 and the young plants were thinned out on Nov. 4. After four weeks a considerable difference in favor of the iodine plants was noticed. On each plot (20 square meters) were grown 60 plants, which were harvested on Dec. 24. The results are as follows :— 0.5 KJ. 0.05 KJ. Control, Number. 19 23 fe) Large plants. Average periphery + Weight. 5440 ¢. 7500 g. 27708. of roots=9.5 c.m. | Weight of roots, 2370, 3360.5; 1440,, Number, 24 20 15 Middie sized plants. Average periphery} Weight. 4020 g, 4220 g. 3020 g. ot roots = 75 cm, Weight of roots, T3705; 1540 ,, IOIO,, (Number 17 17 25 : : a Small plants. | Periphery of roots; Weight. iP 1960 &, 1980 g. 3110g. 4.7¢c.m, and less, | \Weight of roots. 520,, 510;, 790 55 Total weight of plants. 11420¢. 13700 g. 8900 g. * Pe =. LOO: 4260 ¢. S4iIog. 3240 g. 1 Each plot was manured with 200 g. double superphosphate, 312.5 g. (NH,), SO,, and 312.5 g. wood ash, the latter being given in a highly diluted state ten days later, On the Physiological Action of Iodine and Flucrine Compounds on Agric. Plants. 477 This result shows a very favorable influence of potassium iodid in small quantities on the yield with radish. A calculation as to the outlay and profit is of some interest. KI applied for 20 square meter =3/10.05'°¢; Corresponding for 1 ha 2.5) o. its value ==/0-O07en The increase in harvest per 20sq.m. = 2170¢. root, Corresponding per ha = 1085000 g. ,, 289 Kwamme. its value ==) 2280 yen Hence it would certainly be profitable to apply small doses of potassium iodid to the field; the costs would be however very trifling, if we would substitute the crude ash of seaweeds for the purified potassium iodid!. It might be here also called attention to the interesting fact that the farmers along the coast of Japan apply sea weeds as a green manure with very much success, which very probably is not only due to the small quantities of potassa, nitrogen and phosphoric acid, but also to some extent to the small doses of iodine present. Finally I might point out that it might not be advisable to make an application of iodine compounds every year on the same field, since the iodine might gradually be increased to a point where the stimulating action ceases and a noxious action commences. An application on only every second or third year might therefore be preferable. C. On the influence of sodium fluorid on oats. By K. Aso. In a former article was shown that fluorine in the form of sodium fluorid applied in exceedingly high dilution on barley, wheat, rice, soy-bean 2 Since this ash contains about 5 per mille iodine, 5 Kilo of it would suffice to supply the necessary quantity per ha. 2 Bul. College of Agriculture, Tokyo, Vol. 5. No. 2. p. 181. 478 S. Suzuki and K. Aso. and pea plants, can exert a stimulant action.2 In the following lines another experiment with oats will be described. All the conditions in regard to soil, manuring, time of sowing, kind of seed, the number of shoots, watering and harvesting were exactly the same as in the above described pot-experiment made with potassium iodid éy S. Susukz; hence the reader, is referred in this regard to the introductory remarks of the above communication. Pot No. I received on five days 0.01g. sodium fluorid in 100.c.c. water, No. II. o.oo1g and No. III. o.ooo1g, while No. IV served as control. The applications of the highly diluted solutions of sodium fluorid were made on March 11, April 14, 21 and 28, and May 6. On May 20, it was noticed that the plants of No. II. developed best, then followed those of No. I. There was hardly noticed any difference between the plants of No. III, and No. IV. The color of the leaves of No. I. was a little paler than that of the control plants. On May 209, the number of ears was: No. }: & Now: 4 No. III. 4 Neo: Vi eontrol) 2 The plants were cut on July 6. The straw and grains, unhusked, were weighed in the air dry state with the following result : I | IT Ill. IV 2 coer | a” : Number of stalks. 8 | 9 10 9 Weight of grains, un- ~ | yas ep J husked, ¢., Oe: 24.2 25.5 21.4 Weight of straw, ¢. 50.1 45.6 48.6 45.2 This result undoubtedly shows a stimulant action of fluorine in the proportion of 2.17g. in 10000 kilo soil as in No II, although the differences are here not so large as in the case of the pea, described in a former sulletin. 2 Although the presence of fluorine has to be assumed almost in every soil, it is of spec‘al interest that it occurs naturally in wines from certain countries (Holzman). 2 Cf, Bul. V. No, 2. 2 eee On the Physiological Action of Iodine and Fluorine Compounds on Agric. Plants. 479 D. On the influence of sodium fluorid on radish. Bi eke Asa: Two plots, each measuring 10 square metre, had received during the summer, 0.6g and 0.o6g NaF., and again shortly before sowing the seeds of radish, they received o.8g. and o.08g. sodium fluorid respectively. The manure was the same as mentioned in the above field experiment with potassium iodid. The radish was sown on October 1, and the young plants thinned out on Nov. 4. Towards middle of December a difference in development between these plants and control plants was very plain. The plants were harvested on Dec. 24 with the following results :— a. b. control. (0.14 g. NaF) (1.4 g. NaF) Total weight 79708. 64908. | 3814¢. Weight of the ten largest = roots, S | 20508. 1470g. 617 A stimulating action of considerable magnitude is therefore quite evident and it is of special interest that the smaller quantity .o.14g. sodium fluorid has produced a better result than the ten times larger quantity. The cost of production of the increased amount of radish is to be seen from the following calculation: NaF applied for 1o square metres = 0.14 ¢. 5, corresponding to 1 ha. =) 2AOw: Its cost =) 6.4)Sen. increase ot harvest per 10 sq.m. = 4.156 Kg. » corresponding to 1 ha. tii 4 6G. Its value == J10,6:yen. —~ 30> — On the Chemical Nature of the Oxidases. BY K. Aso. The oxidases are considered generally as kinds of enzyms and indeed various of their properties are in favor of this view. Also the observation of Slowtzoff1 and of Epstein? seem to fully establish the enzym-nature of the oxidases. Recently, however, ¥. H. Kastle and A. S. Loevenhart* des- cribed experiments which seem to indicate a certain analogy between the behavior of oxidases and that of organic peroxids towards certain antiseptics and poisons. These authors conclude therefore: ‘the oxidizing ferments are peroxids, formed when autoxidable substances come in contact with air and these peroxids give up a part of their oxygen to other less-oxidizable substances present in the cell.” “In other words, that the process of rendering oxygen active by the living cell, is probably brought about in essentially the same way that this is accomplished by phosphorus, benzal- dehyd and other oxygen carriers, viz, as one phase of autoxidation.” Further, these authors hold that “the function of hydrogen peroxid in the guaiacum hydrogen peroxid reaction, is to react with some one or more of the organic substances present in the plant or animal extract to form an organic peroxid.” ys Physiol, Chem, 31. S/ewtzoff observed that the action of laccase is proportional to the square- root of its quantity. He considers the laccase as a peculiar protein substance which is not changed by pepsin or pancreatin. In the pure state, it is killed already at 5o0°c, in presence of mineral substance however between 65-70°c. 2 Arch. Hyg. 36. p. 140. Z£pstein observed that the presence of hydrocyanic acid in small quantities prevents the action of oxidase and that after the removal of hydrocyanic acid, the activity of oxidase is restored, 3 Americ, Chem, Journ. XNXVI. No, 6. Dec. 1gor. A482 K. Ase. These authors based their view principally on the follewing special observations : 1. Benzoyl-phthalyl-and succinyl peroxids give directly guaiacum blue with tincture of guaiac. Hydrogen peroxid alone gives a faint blue color and that only on heating with guaiacum tincture. The authors mentioned further, that lead dioxid and manganese dioxid give the blue color. But from these facts certainly does not follow that every substance which can produce a blue color with guaiacum must be of the nature of a peroxid. Indeed lead dioxid and manganese dioxid are no genuine peroxids like barium peroxid is and further we find that not only every weak oxidizing agency (nitrous acid, ferric chlorid, potassium ferricyanid), but also oxid of silver and further quinone may produce the blue guaiacum reaction at once.! Some other observations of these authors are however of considerable interest, namely the bluing of the guaiacum tincture by benzoylperoxid can be prevented by hydroxylamine, and phenylhydrazine. In the same way, also the oxidizing action of the potato juice on guaiaconic acid can be inhibited. However, it might be objected that hydroxylamine and phenylhydrazine might merely destroy the guaiacum blue, while they do not counteract the oxidizing activity itself. Indeed, I have already observed some time ago that the blue color produced with guaiacum tincture by the action of oxidase disappears, when a little free hydroxylamine is added. Also phenylhydrazine (0.5) decolorized the freshly formed guaiacum blue. The inhibitive effect of hydrocyanic acid on the action of oxidase was observed again by Aastle and Loevenhart? after Epstein and also myself had made the same observations. A further observation of those authors relates to the inhibiting action N 100 solution of the thiosulfate was added, the blue color with guaiacum was of sodium thiosulfate. When to 2c.c. of a potato extract 0.5c.c. of prevented. Since other enzyms are not injured by sodium thiosulfate, it was inferred that the oxidase is no enzym proper. But here it might be objected * Quinone produces a blue color also with the tetra paper of Wurster, The common quinone is probably a diketon and not a peroxid as formerly believed (Vittig), * They observed that ‘9 parts prussic acid in 10 million parts of juice very nearly mark the limit of the poisonous effect of that acid on the oxidizing substances in the potato, 1 On the Chemical Nature of the Oxidases, 483 that an oxidizing enzym must naturally contain differently constituted active atomic groups than the other merely hydrolyzing enzyms, devoid of any oxidizing power ;! further it might be objected that the oxidase caused the oxidation of thiosulfate sooner than that of guaiaconic acid. I had observed a year since that the guaiacum blue may easily be decolorized by certain compounds present in some plant juices as, e.g., in that of radish and it requires often a certain excess of guaiacum tincture to preserve the blue guaiacum reaction for some time. Also tannin not only prevents the usual guaiac reaction of peroxidase, but in certain quantities can also bleach out again the blue color after it has made its appearance.? Such facts might also apply to the observation of Kastle and Loevenhart that the onion bulb gives no guaiac blue reaction. I can confirm this statement, but if we precipitate the enzyms of the onion with alcohol firs ‘and thus remove compounds that interfer (allylsulfid?), the guaiacum reactions for oxidase and peroxidase, can be obtained with the aqueous solutions of these enzyms, although these reactions set in here more slowly than in other cases. I might further add that the onion juice shows an unusually strong acid reaction and that after neutralization also the guaiacum blue reaction can be slowly produced with the juice itself. Recently Bach and Chodat observed that the juice of Lathraea squamaria yielded on addition of some diluted baryta water, a precipitate which after treating with dilute sulphuric acid produced at once an intense blue color on paper impregnated with starch paste containing potassium iodid.* Their conclusion is, ‘‘ Die sofortige Jodausscheidung aus Jodkalium konnte daher nur von einem acylirten Hydroperoxid herriihren.” 1 A similar observation was made by the writer in regard to the behavior of enzyms toward sodium fluorid ; while most enzyms are not injured by it, the oxidase proper (laccase) is killed easily. 2 Compare my article, *On the Réle of Oxidase in the preparation of Commercial Tea; Bul, College of Agric. Tokyo, Vol, IV. No, 4. p. 256. 8 Schénbein had observed as early as 1864 (Journ, fiir prakt, Chem, Bd. 88.3.460) that certain aqueous extracts of plants give a blue reaction with acidulated iodid of potassium starch, which reaction he supposed to be due to nitrous acid. Many plant juices however yielded that reaction only after standing for a series of days. In the latter case, nitrite might have been produced from the nitrates, frequently present in plants, by bacterial action. 484 K. Aso. But we must take into consideration that iodine can be very easily liberated from potassium iodid by the most different oxidizing influences, in presence of an acid reaction. These authors also observed that some plant juices will lose the property of liberating iodine within a few minutes, If this is so, we have already a clear proof before us that this oxidizing principle is not identic with the oxidase characterized by the guaiacum blue reaction, since this can still be easily observed in a plantjuice after a few days, although the reaction will then be weaker. Also the further interest- ing observation of these authors that in the wilting of a plant the iodine reaction disappears first, militates against the identity of this oxidizing principle with the common oxidase (laccase). I have made a series of tests with the juices of potato tubers and the root of radish, which yield the guaiacum reactions for oxidase and peroxidase very well. But, with these juices, I could not observe the iodine reaction. As I supposed that these juices might contain some substance which interfered with the formation of iodine starch, or absorb the iodine immediately after being liberated I have treated those juices with an excess of absolute alcohol and after washing the precipitates, containning the oxidizing enzyms, with alcohol they were dissolved again in some water. These solutions also yielded the guaiacum reactions upon oxidase and peroxidase very well, but not a trace of the iodine reaction. I applied for one volume of this solution = bes volume of a 2% starch paste to which 1% potassium iodid and 0.5 % acetic acid was added. These mixtures yielded even after twenty four hours standing in darkness, no trace of any blue reaction, while the guaiacum blue reaction even in absence of hydrogen peroxid was still obtained with great intensity.! Bach and Chodat recommend to add some mangano-sulfate in those cases in which the iodine reaction with plant juices fails. But in the above mentioned cases with the juices of potato and radish, this sulfate did not 1 Tn one case I had applied intentionally a potassium iodid solution not freshly prepared, but one which had been exposed in presence of air for a few days to sunlight, In this case, a blue reaction was gradually observed, evidently due to slight traces of free iodine formed in this solution, On the Chemical Nature of the Oxidases. 485 change the result. However after such mixtures were left for a series of hours to themselves a weak reaction set in. But also in some control tests without plant juices, I observed that mangano-sulfate alone in presence of some acetic acid can gradually cause the liberation of some iodine. In order to decide whether the oxidizing enzyms are really organic peroxids, I have made the following experiments relating to the special oxidizing enzym, which produces a red color with a 196 guaiacol solution of weak acid reaction. The juice of the leaves of radish contains besides oxidase and common peroxidase, also a peculiar oxidizing enzym which produces the red reaction just mentioned.! This juice was mixed with > of its volume of a hydrogen peroxid of about 29% and of a faint acid reaction. After five minutes standing, about four times the bulk of absolute alcohol was added and the precipitate very well washed with alcohol This precipitate was then dissolved in some water and tested with guaiacol. but zo reaction whatever was taking place. If Kastle and Locvenhart’s view was correct, then the supposed organic peroxid would be formed almost instantaneously when hydrogen peroxid comes in contact with the proper organic material in the juice. This supposed organic peroxid would consequently be also present in the alcoholic precipitate containing all the oxidizing enzyms, hence the aqueous solution of this precipitate ought to give now without the further aid of hydrogen peroxid, the red guaiacol reaction, but the fact was: zo reaction in absence, but an intense reaction in presence of hydrogen peroxid. What is true for this kind of peroxidase (3-guaiacolase) is very probably also true for the common peronidase characterized by the blue coloration with guaiacum tincture and hydrogen- peroxid,*® but thus far I was not able to prove it in the way just mentioned, , 1 Since Bourguelot observed in the fungus Azssudz, an oxidizing enzym which produces a red color with guaiacol even in absence of hydrogen peroxid, I propose to distinguish this peculiar enzym as a-guaiacolase, from the above-mentioned enzym which T call B-guaiacolase. About this reaction compare also my article, ‘On Oxidizing Enzyms in the Vegetable Body’ Bul. College of Agric. Tokyo, Vol. V, No. 2. p. 207—235. 2 On heating the solution of the enzym precipitate above mentioned for 5 minutes to 75°, the oxidase and the common peroxidase are killed, while the guaiaco!-hydrogenperoxid reaction was still obtained a’though weaker, 486 K. Aso, since I encountered some difficulty in the preparation of a peroxidase precipitate snfficiently pure. It cannot be denied that a transient formation of an organic peroxid takes place when the oxidase causes the oxidation of a certain other compound. Such peroxids are then the first products of an oxidation caused by the oxidising ensym and this opinion seems to be also that.of Bach and Chodat, and differs essentially from the hypothests of Kastle and Loevenhart, according to which oxidases themselves are the peroxids. It must be remembered, moreover, that the liberation of iodine from potassium iodid not only may be due to different oxidizing influences but also that on the other hand, it is not a specific property of all organic peroxids. Thus neither dicthylperoxid nor dibenzoylperoxids will liberate iodine, but denzoylhydroperoxid can do so. But it is a very striking fact that this peroxid can also liberate iodine from potassium iodid in the presence of sodium bicarbonate and not only in presence of free acid. Such hydroperoxids as can liberate iodine are exceedingly powerful compounds,' resembling hypochlorites in their actions ;? hence the amount of such poisons the cells can only be exceedingly minute. Since I have now proved that the iodine reaction does not go parallel to the blue guaiac reaction and since further there exists no proof that organic peroxids are the cause of the iodine reaction in many vegetable objects, it was important to decide the nature of the iodine liberating substance. Two suppositions seemed to deserve some consideration, either there might exist certain organic ferric compounds in some objects or traces of nitrites. The following lines will doubtless prove of some interest in regard to this question. 1 Recently, A. //. Luge (Amer, Pat. 717016 of 30, Dec. 1902) described acetyihydroperoxid : _—@) CH,—C_0_0-H which has a strong odor after hypechlorous acid and has a powerful bactericida action, 2 Compare in regard to the data here mentioned, the articles of Baever and Tidiger in Berichtel der Deutschen Chemischen Gesellschaft, 1899 and 1900, especially in the latter volume, page 1578. ——— On the Chemical Nature of the Oxidases. 487 Experiments with Buds. Sections of potato-buds yielded directly the iodine reaction in presence of some acetic acid.! Also the blue guaiac reaction was directly produced. The cold prepared extract behaved alike. In a second case, with buds from other potatoes, however, the iodine reaction failed, although oxidase, peroxidase and -guaiacolase? were present. Of considerable interest were the observations made with the tubers and buds of Sagittaria sagittefolia. The cold prepared aqueous extract of the bulb gave no iodine reaction, but it gave the blue guaiac reaction, while the extract of the buds yielded directly both reactions. Eight buds of Sagittarta were extracted with 100c.c. water ; a portion was tested directly and another after boiling for 1/, minute. In the former case, oxidase, peroxidase and 3-guaiacolase were easily recognized by their reactions, while in the latter case, no trace of this reaction was more obtained. But very different were the phenomena in regard to the iodine reaction. Not only the unbotled, but also the boiled juice yielded this reaction with great intensity after addition of some acetic acid.* Even boiling for 2 minutes did not alter this result.4. It is therefore undeniable that hereby another proof is furnished that the substance which gives the guaiac reaction for oxidase is not identic with the substance that give the iodine reaction. It was of interest to me to decide whether in the bulbs a compound would be present that can prevent the iodine reaction which so easily and intensely is obtained in the dvds. Hence four bulbs were crushed after removing the skins and macerated with 100c.c. water. The filtrate was mixed with alcohol, whereby a considerable precipitate 1 The tubers did not give this reaction, as was mentioned above. 2 See above p. 485. 3 A blind control test with acetic acid and potassium iodid-strach paste showed no reaction whatever, 4 Bach and Chodat mention that heating to 80°C prevents the iodine reaction, ‘This is, however, probably only the case when the acidity of the juice is more marked than in the case of the Sagttfaria buds. It can then be very easily explained that nitrons acid set free reacts upon amido-compounds and is destroyed with development of nitrogen. 488 K. Aso. was obtained. This was filtered off, the residue washed and after well pressing between filter paper and evaporation of the alcohol at common temperature, extracted with water and tested again. The iodine reaction failed however while the guaiac reaction was obtained. Further tests convinced me that among other substances soluble albumin as well as pepton can prevent the appearance of the iodine reaction which very easily can be understood, since these compounds can bind some iodine, thus rendering formation of iodine starch imposible. Since, the juice of the bulb contained some soluble albumin, it was not surprising to find that the juice of the du/b was capable to prevent the iodine reaction with the juice of the bud, and further that the Jdozled juice of the bulb did not prevent any more that iodine reaction with the juice of the bud. It was further tested whether the juice of the bulb itself would yield the iodine reaction after removing the soluble albumin. But after a few seconds boiling whereby the albumen separated in flocculi, no iodine reaction was obtained in the filtrate, although short boiling does not destroy the active compound as I have mentioned above. The resistance of the active principle towards boiling heat suggested to make a careful test for nitrites and indeed to my great surprise the reaction of Griess for nitrites yielded at once a very decisive result. Hence the liberation of iodine is due not to any enzym nor to any peroxid, but to nitrites. It is very strange that the occurence of nitrite in plants thus far was overlooked. It is true that Schénbeiu more than thirty years ago had supposed the existence of nitrite in plant juices and further that Berthelot had assumed the formation of nitrate in leaves and shoots from ammonia. But some authors did not agree with these observations. The occurrence of nitrite in plants is indeed surprising, since we know that nitrites are very poisonous for plants with an acid plant juice.! But in this regard we must not overlook that the quantity of nitrite present in these shoots is only very small, and that nitrous acid can here not exist in the free state since the acidity of these shoots is exceedingly weak. 1 ©, Loew, Natiirliches System der Giftwirkungen, p. 61 and p, 109. On the Chemical Nature of the Oxidases. 489 Since the question is of some interest whether this nitrite is formed by reduction of nitrate or by oxidation of ammonium salts, I have tested the bulbs with diphenylamine, but no reaction was obtained. The boiled juice of the buds was also poured carefully on the surface of diphenylamine solution in concentrated sulphuric acid, and here soon observed a blue ring, probably due to the small quantity of nitrites present.! A strong reaction for nitrites could not be expected in this manner, since we know that nitrites are in presence of some strong acids and amido-compounds very quickly destroyed with evolution of nitrogen. We can therefore infer that nitrous acid in the buds in analogy to nitrification process is formed by oxidation of ammonia. Summary. It is very improbable that the oxidase and peroxidase of plant juices are organic peroxids. The liberation of iodine by plant juices was proved in one case to be due to traces of nitrite and it is probable that these are present sometimes also in other plant juices. The iodine and the guaiac reactions do not show any parallelism. 1 The reaction of Griess sometimes may be prevented by the presence of certain benzene compounds, like tannins, . Can Sulfo-derivatives of Hydroxylamine Serve as a Source of Nitrogen for Plants ? BY S. Suzuki. It is well known that hydroxylamine as well as diamidogen are not only incapable of furnishing nitrogen to pheenogams but they are directly poiso- nous.1 But thus far no tests have been made with the sulfoderivatives of these compounds. It seemed to me of some interest to test one of such derivatives in this line. I selected the sodiumsalt of a—f8 hydroxylamine- disulfonic acid which preparation was kindly furnished me by Prof. T. Haga.?® This compound has the formula. H ae Na aA), SO; . Na. I prepared the following solutions :— a) 1 per mille Calcium acetate. Boe ee Magnesium sulfate (anhyd.) ger ee Potassium chlorid. Mes Ferrous sulfate (anhyd.) b) 2 per mille Dipotassiumphosphate. oe Sodium a ~ 8. disulfhydroxylamate. and for the control plants : ¢) Same as a). 1 Loew, Ein natiirliches System der Giftwirkungen, 1893, p. 41. 2 This interesting salt was prepared by Prof, Haga from oximido-sulphonate of sodium by a com- plicated process which will be published later by that author. AQ2 S. Suzuki. d) 2 per mille Dipotassium phosphate. O22 Ammonium sulfate. ! The application of two different solutions instead of a single one contain- ing all mineral nutrients was necessary for the following reason. The above named derivative contains two sulpho-groups which on being set free by a decomposition would probably form an acid salt which in itself would be injurious. Hence I applied the secondary potassium phosphate in place of the usually applied monopotassium phosphate. But since that phosphate would precipitate the lime and magnesia of the nourishing solution, it had to be applied separately together with the above named derivative. Barley shoots (about 13 c.m. high) were placed on Nov. 21. in the solutions a) and c), on the following day in the solutions b) and d). This manipulation was repeated in this way for nearly 7 weaks. Two-series with two shoots in each flask were observed. On Dec. 15 the plants were transferred to larger flasks and all solutions renewed. At this time it was evident, that the control plants showed a much better development. From then to Jan. 1oth, a decided starvation was noticeable; all the old leaves died off, while young leaves developed but remained of very small size. The experiment was closed on Jan. roth, since there was no further growth observed and only very few leaves had remained green, while in the control case the plants looked vigorous and healthy. It seems further that the plants can not regenerate hydroxylamine to any noticeable degree from the salt, since a very poisonous action would have soon become noticeable. The state of affairs at the close of experiment is seen in the following table :— Number of Number of leaves. Length of the Weight of the stalks. living dead longest leaves, fresh plants. Solution of a and b series. A { I Ey dees me ae Circ, merce [Go ener | 2 3 6 tTy 8 | I PR rey Pee ee ae Oreiaivapraen 12. HyieG) ae | 2 eee fic SW 8 A eae Pi facneeeee Lr as, | 1 This ammount of ammonium sulfate corresponds to that of the soliam salt of a B. hydroxyl- amine disulfonic acid, in the solution (b), in regard to the amount of nitrogen, Can sulfo-derivatives of hydroxylamine serve asa source of nitrogen for plants. 493 Control plants. I Scere es ee ee Soret ee ee dame ate € | OZ Ss, 2 A EE oe GA Sa ye eee, Rees J I Oi kegs a tae cs 1S CT Seat 7 A ea ar D ; : r son SAP iss ng Abad Suita eh Qe le ao 15-55 Experiment with fungi. A culture solution was prepared, consisting of 200 c.c. Water. 2 g. Canesugar. 0.4 ,, Hydroxylamine disulfonate of sodium. O22 ueihs FO: GOiteare wie SO... Two flasks, a and b, each containing 100 c.c. of this solution were after sterilisation infected with: a) Penicillium glaucume. b) Bac. methylicus. An infection in bouillon from the same source served as control. After 14 days there was no trace of development noticed in the main flasks, while the control flasks showed luxuriant growth. My general conclusions are therefore :— 1. The a. 8. hydroxylamine disulphonic acid is no direct poison for the barley but it is incapable to furnish nitrogen and hence the plants undergo starvation when nitrogen is supplied in this form.! 2. Development of fungi is impossible, when in the culture solutions the nitrogen is offered in the form of a—~8—-hydroxylamine disulfonic acid. 1 It is of some interest to compare with this result the poisonous character of amido-sulf fonic acid, these Bulletins, vol. II, page 487 (1897). On the Influence of a Certain Ratio Between Lime and Magnesia on the Growth of the Mulberry-tree. Since sericulture is one of the most important agricultural industries in Japan, much attention is paid to the cultivation of the mulberry-tree, and various investigations have been published in relation to it. A short review of some of these may be not out of place. The composition of the ashes of healthy mulberry-leaves was found in average, as follows :! Lig Che 9 Chak SG eee ee 12.02 % ee decane ces ep eee es 31.47% INS (OS Seta gol Re ce ae ee 3.14% ee a 33:15.% IG LO) oe see Stda eee eee en 12.48% aN Aree fetes ui ge ge hE Me a oon ee es 4.64% IL as ea ie eee Praesens siete wake ney vas 0.06% SO EAR Oe SS 1.45% LETS ACT Sh OR ee RLS a re 1.59% On analysis of the bark of the healthy mulberry-roots (var. Nezumigae- shi) collected on Dec. 4., I have obtained the following result: In 100 parts of the crude ash, Jf, ODS fa RSS Se AE Lee 18.48 \acecy, WOUND Reeth Roe tec tt pen in ne gee 9.39 OS © SUE RE iced Cane em ee Re Sere 35-50 [Le Cis oA healt ate em een Sele SR Se SE 7.34 1 Nagaoka: Chemical Tables for Daily Use, p. 84. 496 K. Aso. SO eee oa ee es ee 1.38 Si Oey eee eee Dist ae oe we Stes rae Dame eas 2.40 He .Os ovnd. ostiaick gene aictin te tae 8.73 U. Suzuki! has determined the lime and magnesia content of healthy and dwarf-diseased leaves without however observing great differences ; the diseased leaves contained, like the healthy, from 2 to 4 times as much lime as magnesia, although in most cases, the ratio between these was somewhat ereater in healthy than in diseased leaves. Maeno? observed in mulberry-leaves after liming the soil, a moderate decrease of the percentage of woody fibre and increase of the non-nitro- genous extract; further by applying lime, sodium nitrate and calcium sul- fate, not only some increase of the non-nitrogenous extract, but also of the protein and fat. Since the so-called dwarf-disease (Schrumpf-Krankheit) causes an im- mense damage to the mulberry plantations in Japan, it seemed to me of in- terest to look also into the composition of such soils as seemed especially favorable for the development of the disease, that is, causing such a condi- tion of the plant as would render it more susceptible for that disease. I restricted myself to the determination of those quantities of lime and mag- nesia which are available to the root, and for this purpose I have treated the soil with cold hydrochloric acid (10%) for 48 hours. Our experiments with other plants had sufficiently shown that the ratio between lime and magnesia in the soil has a most powerful influence on the development. My analyses, indeed, have shown that the amount of magnesia predominated over that of lime, which is a very unfavorable condition. In 100 parts of dry soil, LOCALITY. CaO Mg O Odakaramura, (Aichiken) civisctiiseesssoues 0.232 0.332 Jotan Sericultural School, (Kyotofu)............ O.1TS 0.259 Angamura: (Ky Gtota) “5. .,., TE 1.5 As the development of mulberry-roots is very vigorous, it could be assumed that these amounts of lime and magnesia might be assimilable for this tree. I altered the ratio between lime and magnesia in this soil by mixing calcium carbonate or magnesium carbonate with it to reach the following ratios :— Pots Ga © : Mg O a I 3 b I 2 c I : I (original) d 2 : I Cc 3 2 i f 4 ; I The surface of the pots measured 0.0495{.]}m. As general manure served : 7.5 g. N inthe form of ammonium sulfate and 5-6 g. P, O, in the form of potassium phosphate for each pot. These salts were applied in solution. Young mulberry plants (var. Shi- h6zaki) of equal size, weighing 976.7 g.—1014.3 g ec. and of stem-length of D> about 30 cm. were planted on April 21. On June 6, and Sept. 19, the following observation was made :— | : Date. I; I. ITT, IV. \ VI. | } June 6, Leaves Leaves Developed | Control, very small, small, best, | | | Sept. 19. One plant Leaves Developed Less well died, With develoy ed to 7 very W ell developed another the | some extent, nearly, same than V. leaves are as IV. very small, OO Oo oo ee On the Influence of a certain Ratio between Lime and Magnesia- 499 On Oct. 1. A photograph was taken (plate XX VII) which exhibits the great difference of development at once. On Oct. 2 the following obser- vations were made :— No, of |} CaO | Number | Fresh weight of Average Number of total leaves. weight of of Remarks. pot. MgO | leaves. g. one leaf, branches. ; He 8 %, Branches were ; 0.33 BS on 2 very small. 10 05 16 6.8 0.41 4 | LN I 21 9.8 0.45 ai ‘- One branch was IV, 2 30 31.9 1.05 7 longest of all. ni Average development We 3 38 36.4 0.98 | § of branches was here better than in IV. Wile 4 20 13-5 0.68 5 Taking now in consideration, that the plant with the lime factor 2 had the longest branch all that other branches were smaller than with the lime factor 3, which latter had also one branch more, it will be safest to conclude that the best ratio CaO: Mg O for the mulberry tree lies between 2 and 3. If follows further that an excess of magnesia over lime depresses the growth considerably ; the leaves become smaller, but true symptons of dwarf- disease are not observed. ere —— On the Influence of Different Ratios between Lime and Magnesia upon the Development of Phaseolus. BY G. Laikuhara. The knowledge of the physiological functions of lime and magnesia is not only of theoretical but also of practical value, as shown by the recent publications of Loew, May, Aso and Furuta. O. Locw has named the ratio a most favorable for plant development the lime factor, taking the absolute quantity of the available magnesia as the unit. Thus it was found that the lime factor for buckwheat is 3, for cabbage 2, for oats I. I have sought to determine this limefactor for Phaseolus and also to observe whether an increase of the absolute quantities of those bases would have any modifying influence upon the result. Thirty small zinkpots of about two Liters capacity served for this ex- periment. Each received 2,5 Kilo pure quartzsand, mixed with the carbo- nates of lime and magnesia in the following quantities and ratios :— Total quantities of Ca CO, +Mg CO, Ae B. Ce for the air dry sand : 0.05% 0.1% | 0.2% = x |—_—__——— ae I par J 3. I I I il Limefactor : Tl oe: ree = 1 I | Ca O =e So | Ss == ee MgO Ill ahs. cs as I I I IV 85 | oS aS I I I \ 0.33 0.33 0.33, I I 502 G. Daikuhara. As general manure for each pot served :— Kg POs ert serene eee one ee 0.1% KP Oy ee ee eee 0.1% FINO ge ore rt cos ee a Se ee 0.2% NE J 2S, cites oe ee re eee 0.19% Be SO). odie eee ee eee ee 0.001 % On Sept. 9th small plants of Phaseolus, grown in sand, and of equal size, were planted into these pots, two in each. After three weeks a con- siderable difference was _ noticed. In the three series the develop- CaO ment was best, where the ratio ————=- was=2. Mg O The following table gives the measurements taken at this time :— CaO pate A B. G Mg O 3 13.5 IS 13.5 Le ot] ot stem 2 18.5 17.0 15.5 I 14.0 14.0 Lies Cc) 0.50 13.0 15.0 10.0 (@) 33 13.0 _ ——- 2 5.0 6.0 5.0 Leneth of the largest 9.0 7.3 5°55 5 I 78 6.0 55 leaf, cm. 0.50 6.0 6.5 5.0 0.33 0.5 pe” 4 2.0 4.0 3.0 Breadth of the largest 2 4.9 3.5 3:2 I 4.0 3.0 3.0 leaf, c 0.50 28 3.5 2.5 0.33 3.5 — On the Influence of Different Ratios between Lime and Magnesia. 503 : ; ei oe mG This table shows clearly not only an tnflucnce of the ratio of Veo : P poo ee upon the height of the plant but also on the size of the leaves. The best vatio ts here 2: r, at least before the fruiting stage of this plant. The plants No. V of B and C had died at the time the measurements were made, very probably from the excess of magnesium carbonate. Unfortunately the experiment had to be terminated soon afterwards on account of fungi making their appearence on the leaves. On the Behavior of the Phosphoric Acid in the Soils Towards Different Organic Acids. BY G. Daikuhara. Many investigations have been carried out to determine how much phosphoric acid in a soil is available for the plant roots. Nearest to the truth came B. Dyer who published an elaborate investigation on ‘ The Analytical Determination of probably available Mineral Plant Food in Soils,” and proposed to apply a solution of 19 citric acid to determine whether a soil is in need of phosphatic manure. He suggested that “when a soil is found to contain as little as about 0.01 percent of phosphoric acid soluble in a I percent sclneoin of citric acid, it would be justifiable to assume that it stand in immediate need of phosphatic manure.”’ I believed to be of some interest to compare citric acid with other acids in this regard and also to compare different soils. I. Application of organic acids in 1 per cent solution. The samples of soil were taken from the experimental paddy field (sandy loam) of the Kinai Branch of the Imperial Agricultural Experiment Station at Kashiwara, Osaka, which were manured every crop with different quantities of phosphoric acid during three years as follows :— No. I. No. P,O, Upland G 2 ae Notes, 37.5 isp Poe as Superphosphate per ha. Now Tx 93.75. Ke P.O. 5, ss er No: 1.) “No: PLO] Paddy : : : Soil No, II. 37.5 Kg P,O, as Superphosphate per ha, S s~“5 » ory 2 Journ, of the Chem, Soc., London, 65, 115 (1894). 506 G. Daikuhara. Each plot had received moreover 112.5 Kilo N as NH,Cl and 93.75 Kilo K,O as K, COQ, per ha. The extractions were carried out according to B. Dyer’s method. The following table shows the result of analysis in the percentage of dry fine earth :— PO, Soluble in oee—\> eee 1% acetica. 19% tartaric a. 19% citrica. 1% oxalic a. /, Nos Ga Non kt Or 0.00832 0.04190 0.08381 0.12955 Upland | = i Z ie Soil. | No. Uh 375 ee Ese Ce 0.04798 0.09596 0.16929 | No. Ill. 93.45KgP,O, 0.01184 0.06978 0.09916 0.18901 7 aNOsee Io Now ©z 0.00096 0.00925 0.01823 0.04606 . Paddy |. Le : 13. / No, AT. 37.5 Ke BLO. oor12s 0.01408 ©,02495 0.05342 Soil. | No. Ill. 112.5 Kg P,O; 0.00192 0.01823 0.04526 0.09660 5 The weakest acid was therefore acetic, the strongest oxalic acid. In other cases, however, tartaric and citric acids extracted a little more than oxalic, as seen from the following table :— / » 3 oils towards different Acids. ‘ we On the Behavior of the Phosphorie acid in the %%GFS00'o 9% SLoo'o ‘opty Aaa | “9vA] %EC'O cane we) %SLroro % 6bzo'O 9% gSo00'o %oOL'O oo ee ne ee Dobibero % Stzo"o o% EI0'0 ia | iz é ‘ A Fi : ‘ ie %BbzO'O %oLI0'0 -3 ANIA %goro %ggL1'0 ; % gr60"0 % me ee £o'o Se %oz:0 =i a DES @ Coxe) | %g900'0 | else MB LEO ‘plor a]exXO "p98 ILL “ploe O1tezIT 7, *plov 9109 MI uraiqnyes “Otd OLE seu ul apqnjos oO" d ‘tava ouy 9G g61L Surureyu0s pos ppy Appr z "ypva ouy BPS'TE Burnezu0s pos pray purydy + *KakvD “LULO'T *cueoy Apurg *KoAtTD “tuvoy Apurs *APUTG “UTA TLC | “(Ouvag nYSoLysy " “YOURIET NYOWIYS *L9]OVA vu) | ‘yourig ofuLg "YURI, 0-07, 3 (jros Appeq) zYOUvIg ursy ; (‘ros pueda) MEN Ny pyuLAg wuryy “WUDANTN(] "MONS [e.VUIAD OAYOT, *UO/JLULIOF suOnLyS “1odxa jeasojoory AO STIOS 508 G. Daikuhara. If. LAxtraction of Soils with Organic Acids of Different Strength. The soil serving for these experiments contained 1.635% of hygroscopic water and 0.1727% of P, O; soluble in boiling hydrochloric acid (sp. gr. 1.25). The method of extraction was also here that of Dyer. The following table shows the results :— Of the acids. Acetic acid. Tartar’e acid. Citric acid, Oxalic acid. 0.25% —_—— — _ a ce) 1326% 0.50% a 0.0403 6 0.08557 0.1609 7 1.00% | 0.035576 0.0495 %6 0.0948 76 0.179826 | 2.00% | 0.0499 % 0.0704.% 0.1029 % 0.1954.% 5.00% | 0.0586% 0.cg18% 0.1173,% 0.1964.% In this case the extractive power of oxalic acid for P, O; in the soil was strongest, next in order came citric and tartaric acids, and finally acetic AlGTG): — 30 —— Can Boric Acid in High Dilution Exert a Stimulant Action on Plants ? BY IM. Nakamura. It has been shown by various authors that the soil of certain districts contains small quantities of borates, hence also the plants grown on such soils contained some boric acid. &. Hotter! proved the presence of boric acid in many plants by extracting their ashes with water and transforming the boric acid into its methylic ether which was distilled off. It is especially the fruits in which the boric acid accumulates; in 10000 parts of the dry matter of fruits the amount of boric acid was found to vary from 2,2 to 12,8 parts. Callison? made similar observations. Of some interest is also the observa- tion of Crampton that boric acid occurs in grapes grown in California. A. Herzfeld and E.v. Lippmann have made further observations on the occurrence of not insignificant traces of boric acid in lemons and other fruits. F. Schaffer* observed recently its normal occurrence in wines. Hotter determined to which extent boric acid and borates will exert a poisonous action on plants. When borates are added to the amount of one per mille to culture solutions, the growth was very much injured and the plants died after 20 days. Even an amount of 1o milligram boric acid per liter can exert some noxious action; some difference in resistance power was, however, noticeable with various plants.5 The source of boric acid in the soils is probably turmalin which mineral contains about 10% of boric acid K Jahresbericht f. Agricultur Chemie, 1890, p. 203, also Zeitschrift fiir Nahrungsmittel, etc. 1895. 2 Jahresbericht f, Agricultur Chemie 1890. % Jahresbericht f, Agricultur Chemie 18809. » Schweiz. Wochenschr. Chem, Pharm, [1902,] 4, p. 478. 5 In regard to algae (Spirogyra, Vaucheria) Loew mentions in Flora, 1892, p. 374 that they are not injured within several weeks by adding 0.2 per mille boric acid to the culture water, 510 Can Borie Acid in High Dilution Exert a Stimulant Action on Plants 2 and frequently occurs in crystalline rocks and granular limestone. Since poisons exert a less powerful action in the absorbed condition in the soil than in the dissolved state in a culture solution, and moreover since poisons in small doses can exert a stimulant action, I observed cultures of barley in soil to which I added 10 milligr. and 50 milligrams respectively of borax per Kilo. These pots were manured with 1g. NaNO3, 1gr. K2CO3 and 1,2 g. double superphosphate. Ten young barley shoots were planted, October 24, into each pot and after the young shoots had reached about 15cm they were reduced to 4 of nearly equal height. The pots were kept in the glass house in which even in the late autumn the temperature on bright days reached sometimes 25°C. Measurements of the shoots to the tip of the longest leaves were made on Nov. 9, Dec. 1 and Dec. 12 with the following results: Measurements Cm, Nov. 9 | Dec. 1 Dec. 12 a | = > RA SASiscacr 31 | 39;¢ 45 Dice seat 27 | 35,5 42 | 5O ng 7 5 | 2 ges Dia Oey ces 26 34 4! [Wola BSR By, A iisaiseiveaedes 28 35,5 46 Average | 28 36,8 43,5 Peet PL xe 30 4I SI | Bhecgraes 24 37 45 ; Soa deemamee 33 2 I Control's, .,205, 7 id 43 5 We aes Seam 25 37 40 Average 29 3935 46,75 The percentage of increase was therefore from Nov. 9 to Dec. 12 the following. Ly gy ) 35976 ; 50 me Borax: ix, “YN average = 35,55 Ay 1h 0 3 36,5. Of +) 39;9/% M. Nakamura. 51L 1) 41,2% 2 6,6% Cava a ) = average = 38,5 % 3) 3553% | 4) 30,096 A photograph was taken on February 15. It is reproduced on Plate XXVIII and shows that 50 milligrams of borax acted very injuriously on the developement of barley, and even as little as 10 milligrams per kilo soil did some dammage. On February 16 were added 0,5 g. ammoniumsulphate in high dilution to each pot. On March 3 the control plants showed developement of three ears, while even 8 days later there was no sign of ears observed with the borax plants. On April 23 the plants were harvested with the following results, showing an injurious action of even the small amount of 10 milligrams borax per kilo soil. Total wt. Number of Number of Average length grains branches of branches Control 45.0¢ 132 8 63,5 cm 10 mg. Borax 2 See 30 4 eo) 50 mg. Borax 1 768ibs 24 4 46,0 ,, In the following experiments, commenced February 1, with pea and spinach the amount of borax was reduced to 5mg. and img. per kilo soil respectively. 10seeds were planted into each pot and the young shoots were reduced to 4 per pot in the case of the pea. On April 24 the following results were observed : Pea Average length Number of flowers 5mg. Borax per kilo soil............ 62 cm 2 rmg. Borax per kilo soil............ 86,25 » 6 MATER rer ote cacewakesscccsavecseccsssace- 69,5 x 3 512 Can Borie Acid in High Dilution Exert a Stimulant Action on Plants? De eee EEE EEE Spinach i Average wt. of plants Average length of leaves 5mg. Borax per kilo soil........... 10,35 g- ; 38,2 cm Controls cs ee Wee eee ee eee 7,2 Z. 34,0 55 { It will be observed that one milligram of borax per kilo soil exerted some stimulant action with the pea plants and 51mg. also with spinach plants. The high degree of poisonous qualities of borax, which even injures plants in doses of 10 milligrams per kilo soil are certainly unexpected. This is of especial interest at present as, a discussion is now carried on as to the admissibility of borax for purposes of preservation of articles of food. In this discussion the remarkable poisonous character of borax for animals is pointed out. /. Hofmann’ inferred from his experiments with dogs and rabbits that boric acid is ‘“‘ein starkes Zellgift.’ Rost’? observed that the body weight decreases continuously by the use of borax and vomition and diarrhea may result. £. Késter* and also G. Merkel' observed that 1—2 grams of boric acid can produce injuries of the stomach and diarrhea. Such an opinion was also expressed by H. Mayer.’ Onthe other hand Liebreich and Gerlach deny the injurious charactar of borax and boric acid in small doses. However, when we take the highly poisonous character of borax for plants into consideration, we must admit also the dangerous character of borax for animals and man. 1 1D, Med, Wochenschr, 1902, No. 46, 2 Ibid. 1903, February. % Zeitschr, Hyg. Igor. 4 M. Med. Wochenschr. 1903, No. 50, p. Ico. 5 Hyg. Rundschau 12, 1230. It must also be mentioned here that Doane and Price reported that calves fed with milk containing borax lost their hairs. Marryland Agric, Exper, Station Report No, 86. On the Action of Vanadin Compounds on Plants. BY S. Suzuki. Although vanadin compounds occur very rarely in nature, vanadin was nevertheless discovered in the ash of the sugar beet by Ed O. v. Lippmann. Observations on the action of vanadin compounds on plants have not been made to my knowledge. It was, however, very probable that in moderate concentration they would act poisonously. Since poisons, however, often exert a stimulant action when applied in very high dilution, I have instituted a series of experiments. In order to observe at first the degree of poisonous action, shoots of barley (20 c.m. high) were placed in solutions of vanadin sulphate of 19% ; 0.1% and 0.01 per cent. After 5 days the shoots in the solution of 19% were dead, while in that of 0.1% the leaves wilted. But the shoots in the 0.c1% solution were still heabthy even after 12 days. Water cultures in Knop’s solution® were also started to which 1.0 and 0.1 per mille of that sulphate was added. A third experiment was made with a soil culture, 10 mg. of the hypovanadic sulphate being added per kilo soil in one pot, 50 mg. per kilo in a second. A third pot served as control. Each pot contained 10 kilo soil and was sown on Dec. 13 with winter barley, 15 seeds in each, which were reduced to 7 of equal size in each pot on Jan. 20. Jar Dery: y 1 Berl. Ber, Vol. 21, p. 3492. * I applied the bluish green sulphate of commerce, the so called hypovanadic sulphate, V, O, (SO,)5. This salt has a strong acid reaction. 3 Cnly the amount of magnesium sulphate was a little increased. * Each pot was manured with ammonium sulphate 2.3 g., sodium nitrate 2 g., potassium sulphate 2 g., sodium phosphate 2.5 g. an sodium chlorid 1 g, 514 Water culture. in 6 flasks.' a and a, b and b, Barley received ” S. Suzuki. shoots (16-17 cm. high) were placed (Dec. 4) o.1 per mille vanadin sulphate. OOF ys » c andc, served as control. ” The observations made on Jan. 20 were as follows :— Lengh of | Number of stalks Number of leaves Length the longest ea we es ae of the leaves. thick thin living dead roots. ot 16.0 c.m I 5 4 9 2.0 C.m. Ms 15.5 I 3 5 8 15 es iverage 15.8 ,, I 4 5 fe) 1.8 b 21.0 C.m., 5 3 22 2 20.0 ¢.m by 210" "5; 5 I 7, 3 20:0. 5 average 21.0 ,; 5 2 20 5 2010) 5 c 18.0 c.m 6 2 21 3 17.5 ¢.m es 22: Onrs 4 2 15 3 LOO average 20.5 355 5 2 1s = 13.8 A very weak stimulant in the case b and b 1° Remarks, \ No root hairs visible. Development stopped ; fresh leaves appear but the old leaves die off. —,, Normal, | | | Normal | action on the roots seemed to have taken place so decisive that the plants were no longer observed. were made on Feb. Length of arf Number Number 27 with the following results :— Length the longest of thick of living of the leaves, thin stalks. dead leaves, roots. y 32.5 cm, 3S 76 10 25 ly, 30.0 Is 70 5 25 veragt 31-30» 20 1 73 9 25 ( 34.0 cn 21 76 13 30 Cc, ole Ken | Is 66 IO = crag ioe 20 71 12 27 This experiment proves that in a normal water much injured by the addition of 0.1 per mille vanadin But in the cases a anda, the poisonous action was The final observations Weight in a fresh roots upper portion state. 39-7 $: 27-48: Sr S 29.555 38.7 », 27.9 45 37:7 8 2598 42.7 45 Boe ss 40:2 49 27-1 3. culture barley is very sulphate, and further The solutions were renewed on Dee. 20, Jan, 8, 20, 31 and Ieb, 21. —— On the Action of Vanadin Compounds on Plants. 51 wu. that when applied in the further dilution of o.or per mille no decisive stimu- lation takes place, although no injurious action is any more exerted. Soil culture. The shoots above mentioned were measured several times. The observations were as follows :—- Average length of the longest leaves. Average number of stalks, g g z Jans 23° March 27. April to. March 27. April ro. o.1 g. vanadin em. Gm: Gin: Pots): (Sanphate per ) 8.6 26.4 57-7 3 3 10 kilo soil. Oma (iso ey) 8.9 20:3 59.1 3 3 Pot IIT, (Control) ; 9.1 Byn2 62.8 4 4 RoteliVies (\33-) 9.3 Be 63.5 4 4 This experiment plainly shows that vanadin sulphate even in a very small quantities has no stimulating action on barley. ooo —____ Can Potassium Ferrocyanid Exert any Stimulant Action in the Soil on Plant Growth ? BY S. Suzuki. In a former article I have shown that potassium ferrocyanid even in a very high dilution acts poisonously on plants in water culture.t~ The ques- tion, however, seemed to be of some interest whether this compound could exert a stimulant action when incorporated in a small quantity into the soil. Four Wagner’s pots each containing 10 kilo soil served for the experiment. Each pot received as manure :— s\ishrausuoiliti) ASW 3 Ghee Meee eC pea Bree, SUC) OT ES Se ee 2.0): 55 [PORES Sai el Bile eer tees eee Br a SMM POO S MAGeN(EPYSt.) “22. .-.2. sec. 3. +e sea sees neve Ee se Two pots served as control while one pot received o.1 g. potassium- ferrocyanid and another 1 g. Fifteen seeds of barley were sown in each pot on Dec. 13. and the young shoots reduced to 7 of equal size on Jan. 20. After a few weeks a decided difference was noticed in favor of the pot that received 1 g. potassium ferrocyanid. Measurements were made on March 7 and 27 with the following results :— t These Bul. Vol. V, No. 2. 518 S. Suzuki. Average length of the longest leaves. Average number of stalks, March 7. March 27 March 7 March 27, Pot’ £ (o.1.gK, Fe €y.)>| 14.4 Cm, a1. fea: | 5 5 Pot Ti: (re. RpReCy,)) |) §s68 5 = | eet. 4 5 Pot III. (Control). 12:3, 37-2 9 4 + Rot DVie (ine. h): nS (oh 5 BS lees, 4 4 The question arised whether the favorable effect in pot II was due to the potassium ferrocyanid as such or to the nutritive action of its decompo- sition products. It was possible that the soil bacteria decomposed the salt, whereby the iron was liberated as ferric hydrate,! nitrogen as ammonia and potassium as carbonate. In order to decide this question 20 g. of the soil of the pot II were extracted on March 17 with water and the filtrate tested with ferric chlorid, but only an exceedingly feeble reaction was obtained. In a second test diluted hydrochloric acid served for the extraction, but with no better result. A control test with 100 g. unmanured soil moistened with a dilute solution of 10 m.g. potassium ferrocyanid showed further that this small quantity is entirely absorbed. It remains therefore for the present undecided whether in the case above mentioned the potassium ferrocyanid acted favorably as a stimulant or by the products of its decomposition as a source of nutrients. ¥ Previous experiments had shown that a small addition of ferrous sulphate to the soil in question increases the yield of rice and of oats, Are Soluble lodids Absorbed by the Soil ? BY S. Suzuki. My experiments on the stimulating action of potassium iodid on agri- cultural crops! made it desirable to know whether the soil can retain iodids in a certain measure by absorption. In regard to chlorids, absorption by adhesion has been observed by various authors. The interesting experi- ments by B. Dyer? on the field of Rothamsted, ¢. g., have shown that chlorids are to a certain degree retained by clay soils. He writes: ‘Now the average quantity of chlorine which falls annually in the rainfall at Rothamsted, as calculated on observations for 22 harvest years, 1877-1878 to 1898-99, was 14.75 pounds.” ‘ Yet we sce that the soil of plat 5 in the Broadbalk wheat-field retains, on the average, within cach depth of 9 inches down as far as go inches, a quantity of chlorine equivalent to that which falls upon its surface each year in the form of rain.* In other words, down to a depth of go inches the soil, though continually subjected to the wash- ing influence of the rain, contains a quantity of chlorids equivalent to that falls upon it during ten years, neglecting the very few pounds annually sup- plied to it as impurities in the mauures.” “It would seem that the clay enters into some sort of combination with the chlorids from which they are only dislodged by a very free application of water.” “ The difficulty of 1 These Bulletins Vol, V. No. 2 and p. 474 in this number. 2 Office of Experiment Stations, Bul, No. 106, U. S. Depart, of Agric. p. 82 and 83. % These quantities are in certain countries comparatively large, Thus in Bardades were found by Albuquerque per million parts of rain water from 6 to 38.5 parts of chlorine, while the nitrogen as am- monia varied between o.or5 to 0.212. (Report of the Agric. work in Barbados, Government Exp. Station, 1902.) 520 S. Suzuki. removing chlorids from the soil by percolation except when a relatively very large quantity of water was used, was demonstrated in some experi- ments described in the paper on the rain and drainage waters at Rothamsted.” In my experiments with potassium iodid I compared “the behavior of this salt in the soil with that of potassium chlorid, 1 per mille solutions of both these salts serving for the filtration through the soil. As reagent for iodine served starch paste to which freshly neutralized hydrogen peroxid and a trace of ferrous sulphate was added. By this reaction of Schénbein very small traces of iodine can be discovered, in the form of the blue iodine starch. First test :— The stratum of soil was 8.5 c.m. high and 5.8 c.m. wide. 200 cc. of each solution were poured gradually on the surface of the soil contained in a cylindrical vessel. After 35 minutes the first drops appeared at the lower end. While now in the case of potassium chlorid already the first 2 cc. showed a moderate and the second 2 cc. a considerable reaction for chlorine? with silver nitrate, there was no iodine reaction obtained in the first 25 cc. of the filtrate. After this a moderate reaction appeared in the next6 ce: and a strong reaction in the following 2 cc. Second test:—Here the column of the soil was higher, namely 15 c.m., but the diameter was smaller than in the former case, namely 3 c.m. While the weight of the fine soil used in the first case was 200 g., it was here only 84g. The solutions were added in this case in such a manner that the surface of the soil was constantly covered by it in a height of 2c.m. The total quantity of solution added was too cc. After about one hour the first drop appeared at the lower end, and while the chlorine reaction was obtained with the first 2 cc. there was no iodine reaction noticed in the first 15 cc. After this the next 3 cc. showed a weak and the following 3 cc. a strong reaction for iodine. Both tests proved decisively that an iodid is much better absorbed in the soil than a chlorid. The calculation shows for ! A control test was made with a distilled water free from chlorine. ‘The first few ce. of this filtrate showed a weak reaction for chlorine owing to the chlorid already present in the soil, but the tur- bidity was much lighter than in the case of potassium chlorid solution, Are Soluble Iodids Absorbed by the Soil ? 521 st experiment that 100 g. soil absorbed 0.012 5 g. potassium iodid and ‘ond case 0.018 g. The effect depends naturally much on the ’ the soil stratum. ' * BULL. AGRIC. COLL. VOL. V. PLATE XXIV. Fig IJ Fig II Fig. I. Agar-plate from the digestive juice of a silk worm 30 days at room temperature. Original plate, Fig, II. The same. Second dilution. BiUEL AGRIC COLL. VOL. V. ELA TIE XV, Plate showing the stimulating action of rubidium chlorid upon barley. I Rubidium plants. II Control plants. To page 464. PLATE XXVI, FMEA ACS (ADILIE. IO EV De ML Ib, II] f iodid of potassium on oats. I, [I] and III, Todine plant s; IV, Control plants. Plate showing the influence ¢ l'o page 474. XXVIII. v7) PLATE WAO a, I BIIKCD (CLOVE TE, TROUT EP AG y plants. To page 499. ios of lime and magnesia upon mulber =e t BiULETAGKIC. COLL. VOLE. V: PLATE XXVIII. I Il III Plate showing the injurious action of borax on barley. I, 0.05 g borax per Kilo soil. II, 0.01 g borax, III, Control. To page 511. TH E BULLETIN OF OTHE SOMmeEGE OF AGRICULTURE, TOKYO IMPERIAL UNIVERSITY, JAPAN. VOL.) VI. 1904—1905. CON TENTS, OF VOLUME VI. Soe On the Wax-producing Coccid, Ericerus pe-la, Westwood. By Prof. C. Sasaki. - On the Feeding of Silkworms with the Leaves of Cudrania triloba, Hance. By PiGiwe.podeakios = Va) 2 SH ee Se eee ee ee ‘Corean Race of Silkworms. By Prof. C. Sasaki. - - - - - --- - - - - The Beggar Race (Kojikiko) of Silkworms. By Prof. C. Sasaki. - - - - - - Double Cocoon Race of Silkworms. By Prof. C. Sasaki. - - - - - - - - On the feeding of the Silkworms with the Leaves of Wild and Cultivated Mulberry- Sitees sey Promeesasakin= =VNefie TeSys Rea ee ee _ Some Observations on ~Anthercea (Bombey) Yamamai, G. M. and the Methods of its-Rearing in Japan. - By Prof..C. Sasaki. .- - - - - - - - - - = /A*New -Field-mousein Japan. By Prof. C. Sasaki. - - - - -.- - - - - “studies-on the Lability of Enzyms. By K. Aso. - - - - - - - - - - - ‘Ueber fungicide Wirkungen von Pilzculturen.. Von Y. Kozai und O. Loew. - - ‘Zar Frage der Existenz des Pyocyanolysins. Von O. Loew und Y. Kozai. - On the Microbes of the Nukamiso. By Sawamura. - - - - - - - - - - ‘Ueber den Kalkgehalt verschiedencr tierischer Organe. Von M. Toyonaga. - - “On the Influence of Different Ratios of Lime to Magnesia on the Growth of Rice. eee eee om EOE TS Sa a ee Se -On the Determination of the Assimilable Amounts of Lime and Magnesia in Soils. yeeeaiaam t= (= Sa eae e BLIP EY se) Se ee ee ee “Ueber den Einfluss des Mangans apf Waldbaéume. Von Oscar Loew und Seiroku eee ane = eS ee Hl Uhl On the Practical Application of Manganous Chlorid in Rice Culture. By K, Aso. - On the Stimulating Action of Manganese upon Rice, Il. By M. Nagaoka.- - - On the Influence of Manganese salts upon Flax. By Y. Fukutome. - - - - - Can Potassium Bromid Exert any Stimulating Action on Plants? By K. Aso. - - Can Thorium and Cerium Salts Exert any Stimulating Action on Phanogamous IJnigtsyeine ASQ = =) = = = = = = = = - = == = = - = Can Salts of Zinc, Cohalt and Nickel in High Dilution Exert a Stimulating Action OnenenculturasPlants?: By M. Nakamura, -<-- - - - = - - - - Can Lithium and Cesium Salts Exert any Stimulating Action on Phxenogams ? yee Nakada - = = =*- = 5 Se = + = = = = ee On the Stimulating Effect of lodine and Fluorine Compounds on Agricultural lapis.) By K. Asoand.S. Suzgukii- -°- = - = - - - +--+ - > On the Tieatment of Crops by Stimulating Compounds. By Oscar Loew. - - - _On the Action of Sodium Nitro-prussid upon Plants. By Rana Bahadur. - - - On the Behavior of Guanidine to Plants. By I. Kawakita.- - - - - - - - Physiological Observations on Bacillus Methylicus. By T. Katayama. - - - - On the Occurrence of Bacillus Methylicus II. By T. Katayama. - - - - - -_ — - — -_ WwW Ww WH Ww Ww oo Oe A | io PAGF. On the influence of liming upon the action of phosphatic manures. By M. Nagaoka. On the action of various insoluble phosphates upon rice plants. By M. Nagaoka. - On the effects of soil ignition upon the availability of phosphorie acid for rice culture in paddy fields. By M. Nagaoka. - - - - - = - = = = = On organic compounds of phosphoric acid in the soil. By K. Aso. - - - - - On the behavior of the rice plant to nitrates and ammonium salts. By M. Nagaoka. On Different Degrees of Availability of Plant Nutrients. By O. Loew and K. Aso. On the Injurious Effects of an Excess of Lime Applied to the Soil. By S. Suzuki. Is the Availability of Phosphoric Acid in Bonedust modified by the Presence of Gypsum? By T. Katayamay*=*= =) =| =.= = =") =.= Ueber den Kalkgehalt verschiedener tierischer Organe. By M. Toyonaga. - - - Ueber das Verhalten von Fluornatrium zu Blut. By M. Toyonaga. - - - - - On the Flowering of Bamboo. By Oscar Loew. - - - = = = = = = = & Further Observations on Oxidases. By K. Aso. - - - - - - - - = - - On the Large Bacillus Observed in Flacherie. By S. Sawamura. - - = - - - Some New Varieties of Mycoderma Yeast. By T. Takahashi.- - - - - - - Can Nitrite Provide Oxygen in Anzrobic Culture for Bacteria? By T. Takahashi. - On Manuring with Kainit. By S. Suzuki.- - - - - - - - = = = = = On the Influence of Various Ratios of Phosphoric Acid to Nitrogen on the Growth of Barley. .By Rana Bahadur. - - - => =|) 9)=55 =e Can Aluminium Salts Enhance Plant Growth? By Y. Yamano. - - - - - - On the Application of Freezing in the Preparation of Certain Articles of Food. By T. Katayama: = =. - 7-9 =) ==) ==) 2 = ee Notes on the Detection and Determination of Fusel Oil. T. Takahashi,- - - - Is Germination Possible in Absense of Air? By T. Takahashi. - - - - - - 105 215 263 47) 285 35 347 353 357 361 365 371 375 387 403 495 42 429 433 437 439 4 ¢ in ] On the Wax-producing Coccid, Ericerus pe-la, Westwood. BY Prof. C. Sasaki, Rigakuhakushi.. Agricultural College, Imperial University, Tokyo, Japan. WITH PLATES I.—II. The waxy product secreted by the coccid insect, which is well known among us, is vulgarly called “ Chiuhakuro” (Insect’s white wax), and is economically employed for certain purposes. It is usually found on two sorts of trees, viz:—Ligastrum Ibota, Sieb. (Jap. Ibotanoki), and Fraxinus pubinervis, Bl. (Jap. Toneriko) ; but Ranzan Ono mentions in his celebrated work,! that it may also not rarely be found on Ligustrum Japonicum, Thunb. (Jap Nedzumimochi). Mr. Stanislas Julien? mentions in his ‘“‘ Nouveaux renseignements sur la cire d’arbre et sur les insects que la produisent, etc. Extraits des Auteur chinois,” the three kinds of trees (Rhus succedaneum, Ligustrum glabrum, and Hibiscus syriacus), on which the Chinese rear the wax-insects. Further, from the article “ Insects a cire” of the same author,? I quote the following lines :—‘ Les insectes a cire sont d’abord gros comme des lentes. Apres l’epoque appelie Mang-Tchong (aprés le 5 Juin), ils grimpent aux branches de l’arbre, se nourrissent de son suc et laissent échapper une sort de Salive. Cette liquer s’attache aux branches et se change en une grasse blanche que se condense et forme la cire d’arbre. Elle a l’apparence du givre. Aprés l’epoque appelée Tchouchou (aprés le 23 Aout, on l’enléve en raclant et on l’appele alors la-tcha, cést-a-dire sédiment de cire.”’ “ Apres l’epoque appelée pe-lou (aprés le 7 Septembre), cette cire se trouve agglutinée si fortement 4 l’arbre qu’il serait fort difficile de l’enlever. On fait fondre cette matiére, et on la purifie en la passant dans une sorte de filtre en étoffe. Quelques personnes la liquéfient & la vapour et la font découler dans un Vase. Lorsquélle est figée et reunie en masse, elle forme C. Sasaki. i) ce qu’on appelle la cire d’arbre.” ‘Quand les insectes sont petits (Cést- a-dire viennent de naitre) ils sont de colour blanche. Lorsqu’ils ont produit de la cire et qu’ils sont atteint leur vicillesse, leur couleur est ronge et noire. Ils se rapprochent entre eux et s’attachent par paquets aux branches des arbres. Dans le commencement ils sont gros comme des grains de millet et de riz, des que le printemp est venu, ils craissent peu a peu et deviennent gros comme des ceufs de poule. Ils sont de couleur violette et rouge. IIs se tiennent par grappes et enveloppent les branches; on dirait que ce sont les fruits de l’arbre. ‘‘ Lorsque cet insecte est sur le point de poudre, il se forme une coque (litteralement une maison) qui resemble aux loges des mantes qu’on voit sur les Murier. Cette coque s’appelle communément la tch’ong (cire graine), ou La-tseu (cire fils). L’interieur est rempli d’ceufs blanc qui resemblent a de petites lentes. On les trouve reunis par paquets qui en renferment plusieur centaines. A l’epoque appelée li hia (le 6 de Mai), on recueille ces ceufs, on les enveloppe dans les feuilles de gingembre, et on les suspend a différentes distances aux branches de I’arbre a cire......... fi The work of Mr. V. Signoret* has proved a great convenience to my study of this interesting subject; but the descriptions of the female coccid as well as its life history appears to me imperfect on certain points. In 1876 Mr. Daniel Hanbury® made the following statement on the coccid: “ Chung-pih-lah ; Chinese insect wax ; pun-tsan, Fig. 837 secreted by Coccus pela, Westw. upon the branches of Fraxinus chinensis, Roxb., which is cultivated for the purpose, and possibly upon other trees, some accounts of the habits of the insect by a comptent observer are much required, the Chinese statements on the subject being extremely obscure.” Further Mr. S. Uyeno® has published some account of the coccid, chiefly extracted from the Report of the English consul at Tchong-King-Fou in China. A brief extract with regards to the coccid is as follows :— “ The wax secreted by the coccid is collected in June, and from the mixture of the former with the fat of the ox, the Chinese prepare candles.” Mr. K. Minemura, who has travelled in Sichuen in China, last year, has brought me some specimens of the coccid and its food plants as well as the att Pies On the Wax-producing Coccid, Ericerus pe-la, Westwood. 1S) transporting sacs for the insect used for the purpose of propagation by the Chinese. On comparing the specimens with those of our indigenous species, we could not find any difference between the two; but the food plant in China is Fraxinus chinensis, Roxb., while in our country, both Fraxinus pubinervis and Ligastrum Ibota, Sieb. are known to serve as the food plants. It s said that the Chinese cultivate the plant specially for the purpose of breeding the coccid, and at a proper season, they carry the female to various localities, where the plants are either cultivated or growing wild, for the purpose of collecting the wax. The wax harvested in a year, amounts to 600,000 Chin (100 chin=nearly 88 yen*). For the last four years, I have devoted some time to the study of this interesting wax-producing coccid (Ericerus Pe-la Westwood), specimens of which have been collected by my friends Messrs. M. Shimidzu, Y. Miyoshi, T. Tsuchida and also by myself, at various localities of our main island as well as in the island of Shikoku. In the following lines, I shall state the results of my study and observations made on the coccid collected from the stems or branches of Fraxinus pubinervis, Bi. planted on the ridges separating rice fields in a village named Okudomura in Chibaken, not far from the city of Tokyo. Female coccid :—The full grown female coccid is pretty large, nearly globular in form and either found solitary or in aggregations; in the latter case, it is more or less deformed naturally by mutual pressure (Fig. 1. PI. I.). The diametre of the largest specimens measures about 11 mm. and the height about 9 mm. (Fig. 2. Pl. I). The dorsal part which forms the larger portion of the body, is dark reddish brown in color, while the flattened ventral surface, by which the insect is firmly attached either to the stems or branches, is yellowish white. The posterior end of the globular body is marked with a deep incision. The dorsal surface is marked with a number of lightly colored transverse ridges, which indicate the abdominal segments. Besides these ridges, there may be found, all over the surface, blackish patches of variable size, and irregular outlines (Fig. 3. Pl. I.). Close to the smaller patches, there opens a very fine * One yew equals to 2.5 francs nearly. 4 C. Sasaki. pore, from which is secreted a very fine light greyish yellow filament, a large number of the filaments accumulate on the body and forms a loosely interwoven filamentous covering. Just beneath the pore lies a large oval glandular cell, which may be seen through the skin. The larger blackish patches, which are less numerous than the smaller, bear some light orange yellow roundish spots, and very often, two of these spots are united leaving only a slight constric- tion between them, or in other cases, two of them are united together by a shorter or longer streak running between them. These light orange yellow roundish spots are the seat of secretion of a sticky transparent mucous fluid. Generally the secretion, accumulating into a light orange yellow drop, hangs down from the body of the insect, and finally they drop down on the ground (Fig. 1, a. Pl. I). The covering of such secretion on the body seems to protect the latter from other insect enemies. The sticky secretion bears a peculiar odor, which is very similar to that of the cedar oil. The ventral flattened surface of the insect is almost oval in shape, but its large central portion becomes gradually concave as the eggs are deposited, and finally this concavity becomes deeper and deeper so as to form a large hollow space, wide cnough to protect many thousand eggs. If we remove the insect from the stems or branches the eggs may freely fall off. The scar left after the removal of the insects from the stems or branches, is an oval greyish yellow ring, whose central oval depression is occupied with a white cottony secretion. The eggs are elongated oval, light yellow, with the diametres of 0.432 mm. and 0.216 mm. (Fig. 4. Pl. I.). Male coccid :—Cylindrical, somewhat tapering towards the two extremeties (Fig. 5. Pl. I.). Head nearly triangular, light orange yellow in color; the dorsal surface is marked with a broad greyish brown median band, whose posterior end, becoming broader, reaches the front edge of the occiput. On either side of this broader end are again some large patches of the same color. There are five pairs of blackish simple eyes, which can be seen when one looks at the vertex of the head. A pair of large round ocelli lies on the dorsal, and also a pair of large oval ones on the ventral side, while the remaining three pairs of smaller roundish ones, lie at the — On the Wax-preducing Coccid, Ericerus pe-la, Westwood. ut sides of the vertex between the dorsal and ventral larger ocelli (Fig. 6. PI. I.). The antenne are long and composed of ten segments, and covered with long hairs. The segments are long except the two basal ones, which are shorter and stouter than the rest. The last or terminal segment bears at its tip three digitules. The thorax is large, elongated and broader than the head. It is of the same color as the head; but the mesothorax bears two dark reddish brown broad bands, which lie close to the lateral sides so as to enclose a nearly hexagonal light orange yellow area. The meta-thorax is marked on its side with a dark brownish oblique streak. The meso-sternum, which occupies the larger ventral surface of the thorax is dark brown and hexagonal in shape. Legs are comparatively long, light greyish brown, and covered with long greyish hairs. The first pair of legs lie far apart from the remaining two pairs. The tibia of each leg is nearly twice as long as the femur, and bears a single spine at its distal end. Two pairs of digitules are present near the insertion of the claw on the tarsus. Fore wings are long oval, nearly transparent, but the costal margin is light brown. The posterior edge bears a single small lobe very close to the insertion of the wing. Balancers are long and stout, of a brownish color, and have each three long slender hooks at the tip. Abdomen is of nearly equal length to the thorax, and its anterior segment is closely attached to the thorax by its entire breadth. It is of a light greyish green color, the abdominal spike or the sheath of penis is rather short and pointed. From the sides of the last abdominal segment grow out two long slender snowy white filaments, which are much longer than the body. The length of the body is 3 mm. Expansion of wings is 5 mm. The male insect appears from the end of September to the beginning of October. They fly about the young female coccid, which is already attached to the stems or branches, and copulation is effected by projecting the abdominal spike beneath the body of the female from outside. After copulation, the male soon dies. Metamorphosis of the Coccid. The female coccid begins to lay eggs from the first part of May, and the young larve begin to hatch out at the begining of June. 6 C. Sasaki. The newly hatched larva is light orange yellow, long oval and depressed (Fig. 7 and 8. Pl. I.). It is about 0.61 mm. in length and 0.37 mm. in breadth. The body is composed of eleven segments. The antenne are short and stout, and composed of eight segments instead of six as described by Mr. V. Signoret+, and all the segments except the single basal one, bear a few long fine hairs. The first and second segments are short and stout, the third is about twice as long as the second; the fourth and fifth are nearly equal in length, and exceed not more than one half the length of the third segment. The remaining three segments, that is the sixth, seventh, and eighth, are much smaller than the others, and the seventh segment possesses an excessively long hair. Eyes simple, roundish, brownish red. The rostral sete, forming an elongated loop on the ventral side of the body, lie beneath the epidermis of the larva, through which the sete can be seen very distinctly. | fhe sthoeraeic as well as the abdominal segments are distinct, but the boundary line between the throax and abdomen is indistinct. The three pairs of legs are of moderate size, and nearly equal in length. The coxa is somewhat stout and long. The femur and tibia are nearly equal in length; but the former is stouter than the latter. A large and long tarsus bears a single claw at its end. At the insertion of the claw on the tibia, there lie two pairs of digitules. The segments of the legs except the trochanter and coxa, bear a few simple hairs. The last abdominal segment is marked with a wide indentation, within which lies, dorsally at each side, a membranous blunt swelling bearing a single long filament. Between these swellings, opens the anus around which are six stout hairs while beneath the indentation, there lies a small semicircular plate, which seems to be the rudiment of the last abdominal segment. The larve distribute by crawling about nearly every young branch, and after moulting passes to the 2nd stage of growth. The distinction of the sexes probably appears during the first stage; but I have failed to recognize it. Male Larva of 2nd stage :—Body oval, depressed, pale greyish brown, with a wide indentation at the posterior end (Fig. 9. Pl. L.). It is about 0.70 mm. in length, and 0.42 mm. in breadth. The dorsal surface of the ; ' ——E———= — OC OO On the Wax-producing Coecid, Ericerus pe-la, Westwood. 7 body s densely covered with snowy white, entangled filaments secreted by the dermal glands (Fig. 10, a. Pl. I.), while the periphery of the body is provided with a row of sharply pointed transparent spines of variable length. Within the indentation or cleft at the posterior end of the body, lies a fleshy protuberance on which the anus opens. Dorsally at the base of this fleshy protuberance, lies a pair of nipple-like appendages each having two small spines at the tip. The roundish, dark brownish red eyes lie ventrally close to the lateral margin of the head. The rather short antennz, which are composed of three segments, are provided with a few long hairs, and lie also ventrally on the head just in the wide space between the eyes. The rostral setae, which have now become free and long thread- like in form, are deeply thrusted into the bark of the host plant. The legs are all rudimentary, and lie close to the ventral surface of the body by their entire length. The first pair of legs, which lie far anteriorly close to the insertion of the rostral sete, is wide apart from the remaining two pairs,- which lie very close to each other. In the last part of August, the male larve (of the 2nd stage) is com- pletely imprisoned within an oval cocoon formed by snowy white filaments (Fig. 10, b. Pl. I.) secreted by the dermal glands as mentioned before. Usual- ly a large number of the oval flattened cocoons lie in irregular masses or completely surrounding the stems or branches, thus forming snowy white patches or a sort of broad girdle (Fig. 12. Pl. II). These white masses of cocoons are again covered with white long and flossy filaments. Within the cocoons, the larve undergo the second moulting, and pass to the third stage. Male larva of the 3rd stage :—Body oval, depressed, light yellow, with a shallow indentation or cleft at the posterior end (Fig. 13, 14; 14, a. and 14, b. Pl. II.). Dorsally the segment lines of the body are conspicuous, and along either side of the median line, runs longitudinally a dark purplish brown wavy band, which meets its fellow at both ends. Anteriorly the unit- ed band soon divides into two broad branches, which again subdividing into two, run far forwards to the anterior margin of the body. Eyes small, greyish, and lie wide apart at the front edge of the body. Antennz lie ventrally at the front end of the body, and is composed of nine more or 8 C. Sasaki. less stout segments, bearing sparsely some long hairs, there being four or five of them on the terminal segment. The rostral setz are long, filament- ous, and of a dark brownish color. Legs moderately long and are com- posed of five segments. The trochanter small, nearly triangular, and closely attached to the side of the proximal end of the femur. At the insertion of the claw on the tarsus, there are a pair of long digitules. The first pair of legs lie far in front nearly at the sides of the insertion of the rostrum, while the second and third pairs, Jying close to each other, are widely separated from the first. About the beginning of October, the larvz of the third stage change into an elongated, dull greyish brown pupa, with a light colored abdomen the ventral surface of which is light yellowish green. Antenne, wings, and legs are all free. The length of the body is 2.2 mm. A few days after remaining in the pupa state, the winged insect appears through a slit-like opening at the free edge of the cocoon. It usually comes out at the posterior end, which bears two long snowy white filaments (Fig. 15. Pl. IJ.), a large number of the males coming out at the same time. The aggregation of the cocoons give the appearance of their being covered all over with long white filaments. Female larva of the 2nd stage :—I have failed to examine exactly the larva of this stage ; but it is probably very similar to the male larva of the same stage. About the end of August, there may be found many young female coccids, lying in groups on the stems or branches, more or less apart from the groups of the male cocoons (Fig. 12. Pl. II). The female larva probably undergoes only two moultings before attaining the final stage. Young female coccid:—Body oval, dorsally conical, and ventrally flattened. The longer diameter of the body is 1.5 mm. and the shorter .35 mm. (Fig. 16. Pl, II.).. The dorsal surface is light greenish yellow, with more or less depressed punctuations. The exuvize lie excentrically on the dorsal surface in the form of an elongated narrow ridge. The posterior end of the body is marked with a deep narrow cleft. The caudal lobes lying on either side of the cleft are long oval and crimson red in color. The margins of the body are thickened, and are provided with a series On the Wax-producing Coceid, Ericerus pe-la, Westweod. 9 of long transparent spines, whose base is supported by a short, stout, chitincus, greenish yellow process, while the tip ends with a transparent pointed cone (Fig. 17. Pl. II.). The transparent spines, thus projecting radially from the margins of the body, firmly attach the insect to the host plant. The ventral surface of the body is much lighter in color than the dorsal. Close to the front margin of the same surface lie two small blackish eyes wide apart from each other. The central portion of the ventral surface is occupied with an oval depression, in which lies a broad lIcngi- tudinal swelling, whose sides are symmetrically constricted so as to form several paired blunt processes, which indicate probably the rudimentary segments of the body. The rostral setz are borne on an elevation lying at the front portion of the central depression. The antenne (Fig. 18. Pl. II.) lie somewhat apart from each other on the flattened ventral surface just above the front edge of the central depression. They are each composed of eight segments, of which the 1st and 2nd are short and stout, the 3rd much elongated, and the remaining ones are gradually reduced in size towards the end, only the three terminal segments bearing a few long hairs. Legs are small, nearly equal in size, and almost rudimentary (Fig. 18, a. Pl. II.). The first pair of legs, lying at the sides of the anterior swelling, where the rostral setz are inserted, are far apart from the remain- ing two pairs. The tarsus is single, and ends with a blunt claw, at the insertion of which are two pairs of digitules. Two pairs of spiracular depressions lie on the broad even ventral area lying between the central depression and the periphery. They have the appearance of a milky white streak. In January, the female coccid grows in size, but still retains an oval shape, with an elevated dorsal and flattened ventral surface (Fig. 10. Pl. If). The longer diameter of the body is 5 mm., the shorter 3.3 mm. and the height 1.32 mm. The dorsal surface of the body is now light greyish orange yellow, and is covered all over with dark punctuations as well as sparsely scattered short spines (Fig. 19, a. Pl. II.) while its ventral surface is provided with small secretory pores. The abdominal segments are indicated by obscure segment lines. The cleft at the posterior end of the body is deep and narrow, and the caudal lobes are spindle-shaped ol (. Sasaki. and of a dark brownish color. The periphery of the body is much thicken- ed, of a hard and brittle nature, and bears a single row of transparent spines (Fig. 20. Pl. II.), which are longer than in the younger stage. The dorsal skin is provided with scattered short spines, and the ventral with loosely arranged minute pores, whose snowy white dusty secretion forms an oval ventral scale on the bark on which the coccid lies. This scale is marked thickly with radial striations, and also by some dark broader streaks, indicating the position of the spiracular depression as well as the deep cleft at the posterior end of the body (Fig. 21. Pl. IL.). About the beginning of May, the female coccid grows larger, and is now light green, with blackish punctuations over the surface. The body is almost conical with a round base. The summit of the cone is tinged with yellow, and from it four or more yellowish lines run towards the base (Fig. 11. Pl. I.). Later, the coccid becomes mature as shown in the Figs. 1 and 2. Pl. I., and begins to deposit eggs, as mentioned before. Product of the coccids:—The wax of the coccids collected by the Chinese is nothing else than the white cocoons formed by the male larve of the coccid, and the Chinese is said to employ it as a material for prepar- ing candles and several ornamental images. Our people also collect the cocoons at some localities for certain limited purposes. In China, the coccid and the host plants are cultivated largely for the purpose of procuring the wax. In order to breed the coccid, the Chinese collect the mature female in April, and they pack them up in a triangular sac made of a leaf of Sterculia sp? (Fig. 22. Pl. II.). Each two of these sacs are tied together by the petioles of the leaves or some other material, and are thus transported to some localities where the breeding is carried on. Some years ago, I have also tried the transportation of the coccid by keeping them in wooden or tin boxes, and was successful. The female as well as the male pup of the coccid are largely infested by a parasitic chalcid fly, apparently of the genus Encyrtis (Fig. 23. Pl. II.). The fly appears in the latter part of August. The females are larger, and are about 2 mm. in length. The body is somewhat long and depressed, and of a dull brown. The head is vertically depressed, and its breadth is nearly equal to that On the Wax-producing Coccid, Ericerus, pe-la, Westwood. II of the thorax. Eyes oval, dark reddish brown. Ocelli light yellow, and arranged on the occiput widely separated from each other. Antenne are rather long, eleven segmented, light brown and lie very close to the upper portion of the mouth. The basal segment is long and cylindrical, the remaining ones are smaller, but gradually increase in size towards the end. The 7th and 8th segments are white, the 9th, 1oth and 11th segments dark brown. The thorax is neariy equal in length to the abdomen, and stout. The scutellum:is large and nearly triangular. The fore wings are large and broad in the outer half, the marginal and submarginal veins dull yellowish brown. They are covered thickly with cilia except for a smaller portion of the inner area. A single clear hairless line runs from the naked inner area towards the outer margin of the wing. The larger hair bearing portion of the wing is marked with three dusky brown patches, which run from the front to the hinder margin of the same. The hind wings are clear transparent, and covered sparsely with cilia. The fringes are somewhat longer than those of the front wing. The distal end of the marginal vein ends at a slight projection on the front margin of the wing, on which are three hooks. Legs are greyish brown and nearly equal in length, but a spine on the distal end of the tibia of the second pair of legs exceeds much in size those of the other pairs Abdomen is sessile, subcylindrical, of nearly equal breadth to the thorax, and light yellowish brown in color. The end of the abdomen is abruptly pointed, and the ovipositor appears hardly beyond it. Spiracular hairs on the abdomen are of variable length. The length of the body is 2mm.; the expansion of wings 4mm. Males are very similar in general aspect to the other sex; but somewhat smaller in size, and differ from the latter in the following points:—The body is dark bluish black, antennz colored uniformly light brown; the front wings lack the light brownish patches. The male genitalia are pretty long and project beyond the end of the abdomen. Length of the body is about 1.2 mm.; expansion of wings about 3 mm. As the results of my study on this interesting coccid insect, 1 may conclude that it is the native not only of China, but also of Japan, where it is widely distributed. Further, the food plants differ in the two countries ; 12 C. Sasaki. Ligastrum Ibota, Sieb., and Fraxinus pubinervis, Bl. in Japan, and Fraxinus chinensis, Roxb. in China. List of References. 1. R. Ono, Honzokomoku Keimo Vol. XXXV.P.9. 1810. 2. Comptes Rendus. Tom. X. P. 619.. 1840. aloe: Citry,.624- 4. V.Signoret, Essai sur les Cochenilles. ¢. Daniel Hanbury, F. R. S., Science Papers, Chiefly pharmacological and Botanical. 1876. 6. S. Uyeno, Shina Boyeki Bussan Jiten. 1888. (Japanese.) ——> o> +— Fig. Fig, Fig. Fig. Fig. Fig. Fig g. Fig. Fig. Fig, Fig. ~ . Q rT: SE Ie GR oS On the Wax-producing Coccid, Ericerus pe-la, Westwood. Explanation of Plate I. Aggregation of Female coccid on the branch of Fraxinus pubinervis, Bl.; a, oil drop. Mature female coccid; a, top view; b, side view; c, looked from behind. 1/1. Dorsal view of ditto, Zeiss B/I. Eggs. Zeiss A/I. Male coccid, Zeiss aa/I. Head of ditto. Side view; Fig. 6, a, Top view. Zeiss B/I. Newly hatched Larva. Dorsal view. Zeiss B/I. Ditto, Ventral view. Zeiss B/I. Larva of the 2nd Stage. Dorsal view. Zeiss B/I. Section of the dermal gland of male larva of znd Stage. Fig. 10, a. filaments secreted by the gland, Zeiss D/4, Femal coccid collected at the beginning of May. 1/1. 1A (. Sasaki: On the Wax-producing Coecid, Ericerus, pe-la, Westwood. Explanation of Plate IT. > Fig. 12. Branch of Fraxinus pubinervis, Bl. with the group of cocoons, and with young and mature female coccids. 1/1. Fig. 13. Male Larva of the 3rd Stage. Dorsal view, Zeiss A/I. Fig, 14. Ditto. Ventral view. Zeiss A/T. Fig. 14, a, Antenna; Fig. 14, b. Leg. Highly mag. Fig. 15. Winged males getting out of coccons. 2/1. 16. Young female coccid. Dorsal view. Fig. 16, a. Side view. Fig. 16, b. Ventral view. Zeiss, aa/t. Fig. 17. Long transparent spines along the periphery of ditto. D/I. Fig. 18. Antenna. Fig. 18, a. Left fore leg. D/I. Fig. 19. Female coccid collected in January, Dorsal view. 10/1. Fig. 19, a. Small spines on the dorsal surface. Zeiss F/I. Fig. 20. Spines on the periphery of ditto. Zeiss D/I. Fig. 21. Ventral scale left on the branch after the removal] of coccid. 10/I. Fig. 22. ‘Triangular sac for transporting mature female coccids. 43. Fig. 23. Parasitic chalcid fly (Encyrtis sp.?) 20/1. BULL. AGRIC. COLL. VOL. V~, PLATE 1 Sasaki et Yekoyama del. BULL. MGRIC. COLL. VOL. Vs. PLATE ii. fig./4, a | - Sasaki et Yokoyama del. : On the Feeding of Silkworms with the Leaves of Cudrania triloba, Hance. BY Prof. C. Sasaki, Rigakuhakushi. Agricultural College, Imperial University, Tokyo, Japan. August 1903. (With Plates III and IV.) Some years ago, I tried several times to feed our native silkworms (Race Awobiki) with the leaves of Cudrania triloba, Hance (Figs 1. 2. PI. Ill and 1V.) which has been cultivated in the farms of our college since its introduction from China, but my attempts were then unsuccessful, as no cocoons could be procured. My friend, Mr. Minemura, who travelled in the central part of China last year, has collected for me some eggs of silkworms largely cultivated in Si-chuen in China, where the Chinese feed them partly with Cudrania and partly with mulberry leaves. This year, I have succeeded to rear the Chinese race by the careful aid of my special student, Mr. Y. Tsuji, and was able to procure cocoons. The ‘results obtained by rearing it with Cudrania, are as follows :— First stage: The larve or silkworms of the Cudrania race’ came forth on May 5th at the temperature of 65°—68° F. They were fed with finely chopped leaves of Cudrania, regularly six times a day. The average length of ten largest worms at the end of this age was about 1 cm. Moulting began on May 12th, and ended on 13th. : 2nd stage: On May 14th, all the worms completed the Ist moulting. The leaves of Cudrania were chopped a little larger than in the previous age, and given regularly six times a day. Average length of ten largest worms at the end of this stage was about 2.1 cm. On the 22nd May, the worms began to moult. 3rd stage: On May 22nd, all the worms completed the 2nd moulting. The leaves of Cudrania were chopped still larger than in the previous stage, 1 Ihave named the Chinese race feeding on Cudrania triloba, Hance “ Cudrania race.” 16 C. Sasaki. and given regularly six times a day. The average length of ten largest worms at the end of this age was about 4cm. Moulting began on the 2oth. 4th stage: On the 31st, all the worms accomplished the 3rd moulting or casting. From the beginning of this age, the worms were separated into two groups A and B. To A, the leaves of Cudrania (chopped or entire) as usual, and to B, those of the mulberry (chopped or entire) were given regularly five times a day. On June 6th, the worms of the two groups attained maturity, and commenced to spin cocoons. The average length of ten largest worms fed with mulberry leaves was 7.1 cm., and their average weight 2.6 grms., while that of the same number of the worms fed with Cudrania, was 7.7 cm. and their average weight 3.4 grms. In each of these two groups of worms, one fed only with Cudrania and the other with Cudrania and mulberry leaves, there appeared two sorts of worms differing in coloration of the body as well as of the cocoons. The first sort of worms, which are more numerous than the 2nd and which are taken as the materials for my expriments, is white tinged more or less with a yellowish shade. The markings on the anterior three body segments are light greyish yellow with a pair of dark greyish spots on either side of the median dorsal line of the 2nd segment. A pair of hook shaped markings of a light purplish hue lies on the 5th and 8th segments. The ventral side of the body as well as the legs are light yellow (Fig. 3. Pl. IV.). The second sort of worms, which are less numerous than the first is white, with a faint bluish color; but no trace of yellowish shade. The markings on the anterior two body segments are dark grey. The two pairs of spots on the 2nd body segment, are conspicuously black. Phe“ sre body segment bears no markings. The markings on the 5th and 8th segments are small and indistinct, but are still of a pale purple. The ventral side of the body as well as the legs are not yellowish (Fig. 4. PI. 1V.). The worms of this race, without passing the 4th moulting or casting, which is usually the case in other silkworms, became mature on June 6th, and spun cocoons. On the 23rd and 24th of June, the moths issued from the cocoons and laid eggs as usual. The cocoons are long spindle shaped or elongated oval in shape (Fig. 5. Oa the Feeding of Silkworms with the Leaves of Cudrania triloba, Hance. 17 6.7. P1.1V.). Their length varies from 37 to 43 mm. and their breadth is 15 mm. on the average. The coloration is of two sorts—white and orange yellow. The whitish cocoons (Fig. 5. Pl. IV.) are formed by the worms, which have no trace of yellowish color on the body and legs; and the orange yellow ones, by those having a yellowish color in their body and legs (Fig. 6.7. Pl. 1V.). The cocoons are destitute of a contriction or depression at the middle, which is present nearly in all of our native races of silkworms. The wrinkles on the surface of the cocoons are moderate. The tissue of the cocoons is more or less hard and compact, but in some cases, they are soft and thin. The average weight of 20 empty cocoons (pupa removed) formed by the worms fed throughout with Cudrania is 0.2135 grms., while that of 20 empty cocoons formed by the worms fed with mulberry from the beginning of the 4th stage, that is, after the 3rd moulting is 0.2065 grms. Thus the different meals given to the silkworms after the 3rd moulting, does not affect their health as well as their growth to any larger extent, and moreover, we see from the above that the empty cocoons of the worms fed with Cudrania are sensibly heavier than those partly fed with mulberry. If we compare the duration of the larval period of the Cudrania race of China with that of our native race Awobiki, there is not any great difference between them, although the former becomes mature and commences to spin cocoons already after the 3rd moulting, instead of the 4th, which our native races generally pass through. The following shows the duration of each age of the two races of silkworms :— | Cudrania race (fed only with Cudrania, and | Awobiki race with it and mulberry) Wee pase | ESS dl SCENE SaKce i ceczs.cccasea es cevesvase 8 days 7 days MEIN Rida ds accaxsesentevesseconse 2 a ma 5 ’ REE EPCs Pins tvns oc cs sxensnvseces 9 6 eet eas vanes sic cudwanav ines cess 74s 7 MR tea y wend ccys deus senses eescvas — » gs 32 days 33 days 18 C. Sasaki. From the above table, we see that although the Cudrania race passes through only three moulting, that is, one moulting less than our native races, the duration of each age is more or less longer than the latter, and thus the number of days of the larval period is nearly similar in the two. Accordingly the quantity of the meals they consume is nearly similar in both. The qualities of the average ten filaments of cocoons taken from the two sorts of silkworms fed throughout with Cudrania, and with Cudrania and mulberry are as follows :— Physical nature of the filaments|}Physical nature of the filaments reeled from the cocoons of reeled from the cocoons of silk- silkworms fed with worms fed with Cudrania Cudrania only. and mulberry. Aver. length of ten filaments............ 516 aunes 527 aunes (613,04 metre) (627,13 metre) Aver. weight of ditto at the length of ATOOPEAILI(ESS | Benppcemcsugsucoacdesc Bice sels 0,147 grms. 0,146 grms, WAVED EEE OL GIO 22 .5..ce sea sentient seo 1,96 denier 1,96 denier Aver. number of duvets of ditto ...... Tal 0,5 Aver. number of ruptures of ditto ... 0,2 0,3 It is very interesting to observe that the parasitic maggot (Larva of Ugimyia sericariz, Rond., which does terrible harm to our silkworms) is entirely absent in the Cudrania race fed absolutely with Cudrania triloba, Hance, while that which is fed with the same plant and later with mulberry trees, is more or less infested by this pest. Consequently if we feed the Cudrania race exclusively with Cudrania triloba, Hance, it will be entirely ree from the parasite, and the crops will be superior than if fed exclusively or partly with mulberry. The results obtained from the foregoing experiments may be summariz- ed as follows :— ist. The Cudrania race of silkworms, which passes only four stages instead of five, has nearly the same length of larval period with our native races, and the consumption of the meals is also similar to the atter. 2nd. The quality and quantity of the filaments reeled from the cocoons of the Cudrania race, are never inferior to our native races. On the Feeding of Silkworms with the Leaves of Cudrania triloba, Hance. 19 3rd. Ifthe Cudrania race is fed exclusively with C. triloba, Hance, it is entirely free from the parasitic maggot, which does great harm to our native silkworms. Explanation of Plates. Plate III. Fig. 1. Branch of Cudrania triloba, Hance 1/2. Plate IV. Fig. 2, Largest leaf of Cudrania triloba, Hance 1/1. Fig. 3. Mature silkworm forming a yellowish cocoon 1/1, Fig. 4. = = +» a whitish cocoon 1/1, Fig. 5. White cocoon 1/1, Fig. 6. Yellow cocoon 1/1. Fig. 7. Ditto of varied coloration. 1/1. BULL. AGRIC.COLL. VOL. Vi. PLATE Jil. Yokoyama del. PLATE Ip. r Fi £¥ 4 - Figs,3-7) Yokoyama del. ; Corean Race of Silkworms. BY Prof. C. Sasaki, Rigakuhakushi. Agricultural College, Imperial University, Tokyo, Ja Fune 903. (With Plate V.) For the last two years, I have tried to rear the Corean race of silk- worms, which I have procured from a Corean friend in Japan. The Corean cartons on which the eggs are laid, are soft nearly rectangular pieces of paper of about 24cm. by 17cm. The surface of the cartons on which the eggs lie, is evenly covered with the ashes of mulberry leaves. The covering of the eggs with the ashes, according to the Coreans, protects spontaneously the coming forth of the silkworms during the summer months. In this con- dition, the cartons are kept during winter, and early in spring, some days before the hatching of silkworms, the ashes are washed away from the cartons and hatching then takes place. The worms came forth at the beginning of May, and after passing the 3rd moult, they become mature and commence to spin the cocoons, thus they lack the 4th moult, which most other races of silkworms generally undergo. In rearing the Corean race, I employed the temperature of 70° to 75° F. in the breeding chamber. The length of each of their ages and number o meals given them each day are as follows :— Number of days Number of meals given taken for breeding. Date. Ages. per day. I 7 Ne 1903 I 2 2 Soe was , 7 3 9 ” ” ” 7 4 10 5, ‘ | ~ C. Sasaki. 22 Number of day . Number of meals given taken for breeding. Date. Ages. per day. 5 Ir V. 1903 I 7 6 Eo inst a: ; Zp 7 13 3° bh) ” I 8 T4 SU Ae = fo) 9 Thess il 2 be) MG oss.) 35 ” 6 II eas eee . 6 12 Too ss 5 6 13 HG) ay, Ee 35 6 14 200s » 6 15 Tee ae * 5 16 iy See te ie 3 17 Oe eae ” 5 18 2A es x 5 19 25 yo > 5 20 AS tye ey 7 5 21 oh EE ee + 5 22 28a ae + ° 23 20) ag5ns9 + fo) 24 30! yyy ass IV 4 25 a1. ; a 4 26 TV “ 4 27 7 er 53 4 28 ee lage 4 4 29 ae , 5 3° By ay 4 31 On became mature ZO. gh) iiss imago appeared From the above table, we see that the Corean race, though it passes through only four ages instead of five, which is usually the case with our silkworms, takes nearly the same length to complete larval life, Corean Race of Silk worms. 22 and consumes nearly the same quantity of leaves as our silk- worms. From the Corean race, I have been able to separate five varieties ac- cording to coloration or markings, which shows that the Coreans are satisfied with breeding such a mixed race, and do not care to select the best varieties from them. The five varieties I have obtained so far are :— Ist. Body whitish with a pale bluish shade at the junction of the body segments. Head dull greyish brown; the dorsal shield of the 1st segment light pinkish grey; no particular markings on the 2nd and 3rd segments. The markings on the 5th and 8th segments are pale bluish purple, and those on the former are imperfectly horse-shoe shaped, while those on the latter are simply curved lines. The tip of the anal horn on the 11th segment as well as the free end of the 12th and the free edges of the last abdonimal legs are faintly tinged with a light brownish yellow. The length of the mature silkworms varies from 6.7 cm. to 6.9 cm. (Fig. 1. Pl. V.).. Cocoons are all white. Their shape is very variable; but the normal ones are long oval, spindle shaped, or perfectly globular. The long oval ones have rarely a slight constriction at the middle of their length. Besides these, there are found many double cocoons, which are mostly larger and very irregular and variable in shape, and thev contain often more than two chrysalids. 2nd. Body whitish with a pale bluish shade all over its surface. Head and the dorsal shield of the 1st segment colored normally. The 2nd seg- ment has a short greyish median dorsal line, and two pairs of small greyish markings. The posterior half of each marking of the inner pair tinged black. Besides the markings of the 5th and 8th segments, there is also a pair on the 9th segment. Those on the 5th are nearly oval, pale purplish blue ; those on the 8th, and oth are small greyish yellow dots. The tip of the anal horn and the free edges of the 12th segment and the last abdominal legs are deep brownish yellow. The length of the mature silkworms same as in the Ist variety (Fig. 2. Pl. V.). Cocoons are either white or light green. The shape is also very irregular. Double cocoons are produced abundantly. 3rd. Body whitish with light bluish and yellowish shades, The head 24 C. Sasaki. and the dorsal shield of the 1st segment colored normally. The 2nd seg- ment bears dorsally a broad light greenish band, which gradually broadens behind. At each side of the broader end of this band lies a simple blackish spot. The median dorsal line on the 2nd segment is short and greyish. The dorsal raised wrinkles on the 3rd segment are yellow. The markings on the’ 5th segment are simple and light greyish purple, while the 8th segment lacks any markings; but the 9th segment possesses dorsally a pair of greyish yellow spots. The tip of the anal horn on the 11th segment as well as the edges of the 12th segment, and of the last abdominal legs are deep brownish yellow. The length of the body same as in the Ist variety GEies )- 3492 tae dele’ NA). : Cocoons are deep yellow, and their shape irregular. Double cocoons are very variable in shape. 4th. Body white with light bluish shade. The coloration of the head same as in others; but the dorsal shield of the 1st segment light greyish yellow. The 2nd segment possesses dorsally a pair of large broad blackish patches, whose outer side is occupied by an orange reddish area. The front edge of this area is lined with black, and there is a single tiny blackish dot in the centre. The joints of the remaining segments are embroidered with a dark greyish band, and the joint between the 11th and 12th segments with two bands. Each of these bands except the Ist, 2nd, and 4th, is marked dorsal- ly with two pairs of small blackish dots ; but the 5th segment bears dorsally a pair of black hook shaped markings, whose broader end lies within the band running between the 4th and 5th segments. Again each of the anterior three bands are provided on either side with a single light reddish dot, and each of the remaining segments with the pair of the same. From the 4th to the oth segment, there are one or two dark greyish dots or patches below the spiracle. The free edges of the 12th segment as well as the last abdominal legs are light greyish yellow. The length of the body same as in the Ist variety (Figs. 4. 4, a. Pl. V.). Cocoons are snowy white, and of variable shape; but normally they are oval and without a constriction at the middle. 5th. Body pale bluish white. Head and the dorsal shield of the 1st segment colored normally. The 2nd segment is marked dorsally with a Corean Race of Silkworms. N wu dark greyish broad band, which broadens posteriorly. Beside this band, there lie on the posterior half of the same segment, two pairs of blackish markings. A narrow line lying between the two blackish markings on either side of the 2nd segment is light orange yellow. The dorsal raised wrinkles on the 3rd segment are also light orange yellow. The markings on the 5th segment are oval, and enclose a single blackish spot, and their outer edges are lined with black, while their inner halves are of a light orange yellow. Besides the small roundish markings on the 8th segment, there is also a pair of similar markings on the 7th, and both are colored orange yellow. The free edges of the 12th segment and the last abdominal legs have a light greyish yellow color. The 2nd, 3rd, and the anterior half. of the 4th, segments as well as all the remaining segments have a light greyish shade on the pale bluish white ground color. Moreover, the dorsal surface of the 4th to 11th segments is marked with a broad longitudinal dark bluish grey band, which is lined on either side with a pale blue. On the median dorsal line of the 5th to 1oth segments, there lies, on each, a x shaped pale bluish marking. The length of the body same as in the Ist variety (Fig. 5. Pl. V.). The cocoons of the above mentioned five varieties are equally very variable in shape, and there is no any fixed shape among them; and moreover, double cocoons are more numerous as compared with our white native races. The principal shapes of the cocoons are oval, conical or spindle shaped, but they may be often nearly or perfectly spherical or rarely oval with a slight constriction at the middle (Figs. 6—11. Pl. V.). The color of the cocoons has some relation to the coloration as well as the markings of the silkworms, thus :— the cocoons of the Ist variety are all white ” ” ind PANE hey » White or light green ” ” ah EY 5s » deep yellow » ” ye eels. £55 »» snowy white ” - evil ae » White or light green. The cocoons of the 4th variety are much superior in their brilliancy. The double cocoons, which amount to about 50 % of the total, are irregularly roundish or oval, and more or less depressed. Their diameter 26 C. Sasaki: Corean Race of Silkworms. varies from 23 to 27 mm., and their height from 12 to 18 mm. These shapes of the double cocoons, which are usually not found in our native races, are very peculiar to the Corean race (Figs. 12—15. Pl. V.). The following gives some of the qualities of the cocoons of the five varieties selected from the Corean race. 1st Var. and Var. 3rd Var. Aver. length of 10 filaments (Bave) of cocoons. 363 Aunes. 329 Au. 22 Au. (432 Metre.)| (382 M.) (502 M.) Aver. titres of ditto at 400 aunes, 1.36 Denier. 1.28 D. rarely Aver, duvets of ditto, 1.5 2.0 0.6 Aver. ruptures of ditto, O.I 0.2 0.6 4th Var. 5th Var. 388 Au. 316 Au. (462 M.) (376 M.) As the result of any study on the Corean race, I think, if we spend some time in improving the white cocoons of the Ist and 4th varieties, and the yellow cocoons of the 3rd., it will not be difficult to obtain some excellent varieties of both white and yellow ones from the Corean race studied by me. Explanation of Plate V. Piges et. Corean race Ist variety 1, Fig. 2. Ditto ond! a3 I Big: 23. Ditto Std tsa Big as Ditto Athy a. I Bro. 1G; Ditto RL I Figs. 6-11. Cocoons of Corean race 1/ Figs. 13-15. Double cocoons of ditto 1 ie . ite I. ALE ¥-. MULL. AGRIC.COLL. VOL. V7. 4 VU. fig. ee The Beggar Race (Kojikiko) of Silkworms BY Prof. C. Sasaki, Rigakuhakushi. Agricultural College, Imperial University, Tokyo, Japan. August 1903. (With Plate VI.) Two years ago, I had the opportunity of procuring the eggs of the so- called beggar race (Kojikiko) from Shigaken (prefecture near Kyoto). As the name implies, the beggar race eats voraciously not only fresh and clean mulberry-leaves, but also the withered, spoiled or other waste leaves, which are rejected by other races. Notwithstanding that the meals of the beggar race are of an inferior quality, and its treatment imperfect in every respect, it is still very strong and healthy, and grows well like other silkworms, and moreover, there ap- pear only a few diseased silkworms during its cultivation. However it is very strange that although this race is reared to a limited extent by nearly all cultivators in certain districts as an extra product, it has till now been generally uuknown among most of our cultivators. This race appears twice a year, and its cocoons are yellow, and more or less inferior in quality to the white or green races of our silkworms. The spring breed reared this year (1903) came forth on May 3rd, and matured on the 7th of June. The worms were fed mostly with withered, or spoiled leaves or those which were destined to be thrown away as a waste. The number of feedings each day as well ‘as the ‘quantity given ata time during cultivation, was intentionally made very irregular. The following shows the ages, dates, the number of days required for cultivation, and the number of meals given each day for the spring breed. 28 C. Sasaki. = i Number of days Number of meals per Ages Dates Bee ge 8 ee ee required for cultivation day I 3 V. 1903 I 3 4 5 9» ~ 4 > 3 | 4 wr \ “I | e | 7 3 5 5 > S re, 33 16) 4 | Oia ’ 7 5 TOU) Ass 8 6 Tiles ; fa) 3 I2 ;5 ; 10 Oo 1 IBS Ss II 5 ia 5. 12 4 17 15 6 : IIT, 19 17 2 | 4 On Id 6) 21 ; 19 6 22 Oo 6 23 21 5 DA ee. : 22 5 1\ 25 23 I 7 | 4 e “| “5 5 | | Q | HA } 25 260 5 | 0) 27 5 | Y sy ’ 25 5 | | x1 yy 29 2 . | : | v r Vio to , 30 2 2» , j1 t | | The Beggar Race (Kojikiko) of Silkworms 29 Number of days Ages Dates required for cultivation Vv. Bue 1908 32 ae ‘ 33 Cope 34 6 == 35 Be ets 36 Number of meals per days SSeS On the 7th June 1903, the silkwoms became mature and began to spin cocoons. On the 17th June 1903, the moths appeared and laid eggs. The summer breed came forth on the 4th of July and matured on the 30th. It was similarly treated with the spring breed. The following shows the ages, dates, the number of days required for cultivation, and the number of meals given per day for the summer breed. ——— ee eee Ages Dates Number of days _ required for cultivation ds 4 VIL 1903 ae ate Bees 2 Oto 3 i 4 Shor 2 5 9 » , 6 TOUR Fe ~ ie Piers 7 8 Ele 5 9 13.939) os 10 14 3 II Il. Cee _ TO); Ss 13 NUT ees ss 14 1S! 5 a 15 19 : 16 Number of meals per day Cle eats See ee = 3 i “ mf i) 12) 6 30 C. Sasaki. Ages Dates Number of days | Number of meals = required for cultivation per day iV. 20 VII 1903 17 6 Qe oe, ss 18 6 22s 19 6 23» 29 " 24, ” 21 4 \ 25,55 Wy 22 7 20) We ‘3 23 7 27» , 24 6 2840" ; 25 6 29 » » 26 6 On the 30th July the silkworms matured and commenced to spin cocoons. On the 1oth August the moths appeared and laid eggs. The mature silkworms of the spring breed, which are nearly similar in size and appearance to the summer breed, are about 6.7 cm. in length. There are two varieties of these worms, viz :—1st: Body is white, its anterior and posterior segments of a faintly yellowish hue. Head dull greyish brown, the dorsal shield of the 1st body-segment (thoracic dorsal plate) is light pinkish yellow. The markings on the 2nd and 3rd body segments are entirely absent, those on the 5th is light purple crescent-shaped, and those on the 8th are reduced to merely pale purplish dots. The free end of the last body-segment as well as the last abdominal legs colored light greyish brown. The ventral surface of the body as well as the remaining abdominal legs are also of a yellowish color. (Fig.1. PL. VI.). 2nd.: The color of the head and body as well as that ofthe last body-segment, ofthe last abdominal legs and of the thoracic dorsal plate is similar to that of the Ist variety. At the junction of the 2nd and 3rd body-segments, there lies, on either side, a pair of nearly triangular black markings. The two markings of each pair is separated by a light pinkish streak. The inner marking of each pair is connected together by a blackish streak running transversely on the dorsal surface. The markings on the 5th body-segment are also light purple and crescent shaped and enclose a dark purple dot on their concave side. The The Beggar Race (Kojikiko) of Silkwoms 31 markings on the 8th body-segment are somewhat larger than in the Ist variety and are also purple (Fig. 2. PL. VI.). The cocoons are cylindrical and yellow, having a shallow constriction at the middle (Fig. 3. PL. VI.). The sizes of the cocoons differ in the two breeds, viz: the largest cocoons of spring breed are 2.8 cm. in length and 1.4 cm. in breadth, while those of the summer breed are 3.1 cm. in length, and 1.6 cm. in breadth. Thus, the cocoons of the summer breed are larger than those of the other breed, and the color of the former is deeper than that of the latter (Fig. 4. PL. VI.). The qualities of the filaments and raw silks reeled from the cocoons of the spring breed of Kojikiko are as follows :— Quality of filament taken from a single cocoon. Pvenlenety Giro niaments.... .. .-. .-- 2.95 aunes=351 metre. Aver. titre of 10 filaments for the length of 400 aunes (476 Metres). = 1.35 denier Aver. number of duvets of to filaments for the length of 400 aunes. 0.9 Aver. ruptures of 1o filaments for the length of 400 aunes 0.3 Quality of raw silk. prepared from 6 or 7 cocoons. Aver. titre of 10 samples of raw silks for the length of 400 aunes. =) Lo:2 denier Pvereeenacity Of ditto 7. 4.. %..: -.. ... 40.0 grams Pwervelasticity Of ditto... =... -:. ... ... 10.54 cm. Although the lengths of the filaments as mentioned above are generally less, and the qualities of the same as well as the raw silk of the race Kojikiko more or less inferior to those of our white races, it will be still profitable to cultivate as an extra product on account of its strong resistance to diseases, as one Can procure a sufficient crop without much pain. | Double Cocoon Race of Silkworms. BY Prof. C. Sasaki, Rigakuhakushi. Agricultural College, Imperial University, Tokyo, Japan. August 1903. (With Plate VI.) The double cocoon race of the silkworms is aboriginal to the Riu Kiu Islands, and is largely reared there for the purpose of preparing rude coarse or floss silk ; but not for commercial purposes. As this race is strong and healthy, it can be cultivated by the natives with great ease It appears once a year, and the duration of its cultiva- tion varies from 28 to 30 days at the temperature of 70° to 75° F. in a breed- ing chamber, and thus it matures three or four days earlier than the white races of our main island. There are two varieties of this race ; but both spinn yellowish cocoons. Ist variety :—Body is light bluish white ; head and the dorsal shield of the 1st segment are dull greyish brown. The dorsal surface of the 2nd seg- ment is marked with a broad band of light greyish yellow, whose anterior half has a dark greyish streak in the median line. On either side of this broad band, there are two blackish spots of different sizes. The raised wrinkles on the 3rd segment have a pair of light greyish spots lying apart from each other. The markings on the 5th segment are comma shaped. They are greyish yellow, and are bordered with blackish lines. Within each of these markings lies a blackish curved line. The markings on the 8th segment are round and greyish yellow, the peripheries are lined with black, and a small whitish spot lies at the centre of each. The 5th to rith segments are marked densely with minute greyish dots, and besides these markings, each of the segments bears two pairs of distinct blackish spots. The anal horn and the abdominal legs are tinged yellow. The length of the mature silkworm is 7.6 cm. (Fig. 5. Pl. VI.). 34 C. Sasaki. 2nd Variety :—Body light yellowish white, head and the dorsal shield of the Ist segment colored same as in the rst variety. The dorsal band on the 2nd segment, and the raised wrinkles on the 3rd are light greyish yellow. A pair of blackish markings at each side of the band on the 2nd segment is smaller than in the last variety. A pair of large comma-shaped light grey markings on the 5th segment have each a dark greyish line within. The markings of the 8th segment are imperfectly ring-shaped, and are of a light purplish ashy color. The anal horn as well as the abdominal legs are tinged yellow (Fig. 6, Pl. V1.). The cocoons of this race are almost all double, and the simple cocoons containing a single pupa are much less numerous; while the flossy silk which covers loosely the surface of the cocoons are comparatively abundant than in other races. The simple cocoons are spindle shaped; but they are often deformed. Their color varies from light to deep yellow. The length of the largest cocoons is about 3.3 cm. and the breadth 1.5 cm. (Figs. 7, 8, Pl. VI.). The double cocoons, which are characteristic of this race, are excessive- ly large and very variable in shape; but they are generally hard and com- pact in texture (Figs.9, 10, 11, Pl. VI.). They enclose usually more than two chrysalids, and not rarely even seven or eight chrysalids (Fig. 12, Pl. VI.). The cocoons are mostly ovate-oblong, elongated, triangular and more or less depressed ; but still other forms are often met with. The length of the largest cocoons is 7 cm., and the breadth over 3 cm. ' >a ‘ tar La Double Cocoon Race of Silkworms. 35 i Explanation of Plate VI. Fig. 1. Mature larva of beggar race without markings 1/1. Ris 2) Ditto i eas - » with iif ice Fig. 3. Cocoon showing the coloration, Fig. 4. Showing the sizes of the cocoons of two breeds. a, of spring breed, b, of summer breed. Fig. 5. Mature larva of double cocoon race. st variety. a, dorsal. b, side view. Fig. 6. Ditto and variety. Figs. 7, 8. Simple cocoons 1/1. Figs. 9, 10, 11. Different forms of double cocoons 1/1. ‘ Fig. 12. Double cocoon cut open to show the contained pupce 1/r. BOL. AGRIC. COLL. VOL. Vis. PEATE VI. = ae Sasaki et Yokoyama del. 1-4, Double-Cocoon Reece. 5-12, Begger-Race. On the Feeding of the Silkworms with the Leaves of Wild and Cultivated Mulberry trees. BY Prof. C. Sasaki, Rigakuhakushi. Agricultural College, Imperial University, Tokyo, Japan. August 903. In the year 1coo, I made some experiments on the rearing of our native silkworms (Race Aobiki) with two sorts of mulberry trees—wild and culti- vated,—in order to examine whether or not these different mulberry trees give any effects on the nature of the filaments, which make up the cocoons. The rearings were carried on by my assistant Mr. Y. Bannai and by a special student, Mr. M. Tokunaga, to whom my acknowledgements are due. Before entering into details, I think it will not be entirely useless to mention the methods of plantation of our mulberry trees, of which there are two principal ways, viz.—zegari (cultivated) and ¢akagz (uncultivated or wild). 1. The “ zegarz”’ method is extensively practiced in the north-western districts of our main island, where it is esteemed as an improvement. ? F, bulky, and oval in shape (Fig. 2. Pl. IX.). Incisive foramina is very small but distinct. Of the upper jaw, 1st. molar with 4 closed triangles and an anterior loop; 2nd. with three closed triangles, and a posterior triangle with an closed triangles with an anterior and a posterior open base; 3rd. with 3 3 3 loop, with 4 inner and 3 outer salient angles; of the lower jaw, Ist. molar with 5 closed triangles, an anterior trefoil and a posterior loop; 2nd. with four triangles and a posterior loop, each triangle on the one side confluent with that on the other; 3rd. with 3 long inner and 3 short outer salient angles (Figs. 4; 4,a. Pl. IX.). In general aspects and characters, the present species resembles to a certain degree Arvicola subterraneous, Selys., which is described by Mrs. H. Leunis' and J. R. Bos.*; but it differs in the coloration of the fur, length of the body and tail as well as the size of the ear. This leads us to give it a new name Arvicola hatanedzumi.’ Habits: The field mice are mostly found during winter in the farms of wheat, tea, mulberry trees and other plantations. In the day time, they conceal themselves within the subterranean nests, while at night they come out from their hiding places and search for food. If we feed the mice in an enclosure, they remain quiet during the day, but as night approaches they become very active and emit a peculiar cry. The nest of the mice are usually constructed on the dikes separating rice field or mounds, elevated farms scattered over the same field or along 1 Hf, Leunis, Synopsis der ‘Thierkunde Band I, 2 R. Bos, Tierische Schadlinge u. Niitzlinge. Hatanedzumi (Jap.) means Farm mouse, A New Field-mouse in Japan. 53 road-sides directly exposed to sun-shine; and moreover even on a plain farm on which various stuffs of halms, stalks or roots of the farms are piled up. The nests are constructed in an oval hollow, excavated usually at the depth of five to seven inches below the ground. Its walls are provided with one or more opening, which communicate with tunnels running in various directions and extending to various distances, where the mice may be able to procure their food. These tunnels open at various distances to the surface of the ground by roundish holes, which serve as a clue to the general direction of the tunnels. The tunnels extend as far as where the food plants are to be detected ; and close to the wheat, tea, mulberry and other farms which are preferred by the mice, there open usually one or more holes. In the case of wheat, the mice come out to the surface from the holes opening close by, cut off the stalks or leaves at a height of less than an inch above the ground, and eat them on the spot, or else they carry them to their nests; thus the holes and newly cut stalks or leaves of the wheat indicate, without doubt, the presence of the mice. But in the case of tea and mulberry trees, the mice do injury only to the root by gnawing the cortical layer leaving series of traces of their teeth on the surface of the woody layer (Fig. 5. 15) lB The nests (Fig. 6. Pl. 1X.) are generally oval (length about 22 cm., breadth 14 cm.), or nearly roundish, more or less flattened, consist of a single chamber. The materials employed for the construction of the nests are generally fine strips of straw or fibrous roots of various plants. The inner layer of the nest consists of a much finer and softer stuff than the outer. The nest is provided with the same number of openings as the hollow within which it lies, thus giving the mice free passage towards the tunnels. Close to the hollow in which the nest lies, is excavated another small chamber or hollow mainly used for preserving food. The principal food stuffs, so far as I could find in the store chamber, are strips of the roots of tea or mulberry trees, roots of Lappa major, Gertn., Daucus carota, L., the stalks or leaves of the tobacco plant, ears of the rice plant and others. The roots are cut off nearly to an equal length and piled up horizontally in a regular manner in the store chamber ; and especially the ears of the rice plant are equally cut off to the length of four to five inches, and then they are piled up 54 C. Sasaki. horizontally by arranging regularly the grain bearing end of each ear on the same side so as nearly to fill the chamber. Generally a single nest is constructed at one spot, but sometimes more are found, The interior of the nest is always very clean and free from dusts or excrements. On the surface of the ground below which a nest lies, usually open one or more holes, and in the case of the nests formed beneath the inclined surface of dykes, boundaries &c., the holes are not far removed from the nest, and open always more or less below the level of the nest so as to avoid the entrance of rain water. If the nests are constructed below the inclined surface as stated above, the particles of soils will flow out from the holes on the same surface. When the particles of soils look fresh, the mice are almost always present in the nest, while if not fresh they are usually absent, thus we can easily judge of the presence or absence of the mice by the appearance of the particles. In winter, there may be found several individuals in a single nest ; but during the breeding season, it is most probable that a single pair inhabits a single nest. In capturing the mice, if we dig out the nest slowly, it is always very difficult to find them, for by their acute sense of hearing, they will soon notice the approach of men, and escape through the tunnels running out from the nest. But after having located the nest, let several persons dig out the ground around the nest at the same time at a distance of a few feet from the latter so as not to allow their escape through the tunnels, then the mice can be easily captured. The pairing season of the mice is not yet accurately known, but they seem to breed several times during the warmer months of the year. They are herbivorous in habit, but when starved they do not hesitate to devour their weaker and inactive mates. Bigs Fig, Fig. Fig, Fig, Fig, Fig, Fig, Fig. A New Field-mouse in Japan. 55 Explanation of Plate IX. (Figs sc 1; I,a, 6, drawn by K, Yokoyama) 1. Arvicola hatanedzumi, Sasaki 1/1. I,a. Ditto. 2. Skull of ditto . 1/r1. 3. Right fore foot r/r, 3,a. Right hind foot 1/1. 4. Molar series of right upper jaw 4/1. 4,a. Ditto of left lower jaw 4/1. 5. Root of mulberry tree with the trace of teeth ; a blackish line shows the level of ground. 6. Nest with two openings 1/3. sv VIN/SN (GN AN ‘\ ‘bh ; AS V V (AYN th aval ») (x, 74 /\\ tb > I P, WAIN) ( VI. VOT. AGRIC, COLL, BLL. — Studies on the Lability of Enzyms, BY KK -Aso; The cause of the chemical powers of enzyms has frequently been the object of speculation. According to the theory of O. Loew,! this activity is intimately connected with the lability of the enzyms, in as much there exist in them certain labil groupings which exert chemical energy—a kind of atomic motion—which can cause chemical changes in certain other compounds. A condition for the action of enzyms is that the compound to be acted upon, shows a certain configuration as was shown by £. Fischer. . Various compounds can destroy the activity of enzyms what can be explained by their causing the migration of atoms from the labil to the stable position within the enzym molecule. But, in most of such cases no conclusion can be drawn as to the nature of the labil groups. Thus, for instance, carbonate of soda in 1 per mille solution will soon destroy the action of pepsin and takadiastase. This is a special case of the phenomenon that alkalies and acids can change various labil compounds to stable ones. In order to be able however to draw certain conclusions as to the chemical nature of the labil groupings we must select such compounds that have quite specific actions, even in high dilution and in perfect neutral solution. Loew suspected formerly that the lability of and aldehyde enzyms is caused by the simultaneous presence of amido groups, but his own tests with alkaline silver solution failed.2 The presence of aldehydegroups would provide a plausible view, as Vernon* 1 Pfltig. Arch. 27., 212; Die chemische Energie der Jebenden Zellen, p. 149; Journ. f, prakt, Chem. 1888, p. 194. 2 Pfliigers Archiv, fiir die ges. Physiologie, vol, 27, p. 212. 3 Journ, of Physiology, vol. 29, p. 331 [1903]. K. Aso. 1 has pointed out: “it may, for instance by alternate hydration into CH (OH), groups and subsequent dehyration be able to effect the hydrolysis of proteids, whilst di-amido-or other aldehyde groups by the reverse process may be able to effect the dehydration of caseinogen into casein.” As to the action of zymogens, Vernon expresses inmself as follows: “Let us also provisionally accept Loew's hypothesis that ferments differ from inactive proteids in virtue of their containing aldehyde groups. Then we may assume that the formation of ferments in the cells of digestive glands consists in the activation of ordinary proteid molecules by the reduction of some or all of their COOH or acid groupings into CHO or aldehyde groupings.” It is also possible that according to a later view of O. Loew, the zymogens contain ketonegroups, and that the activation process consists in the opening of lactamgroups in the zymogen molecule, labil amido- eroups thereby being generated.1_ Amidoketones also are very labil bodies as seen from the behavior of diamidoacetone which spontaneously changes to an indifferent substance (Riighezmer and Meschel). In regard to the amidogroups O. Loew infers their presence from his observation that dilute formaldehyde easily destroys the action of enzyms at the ordinary temperature.? It is well known that formaldehyde easily attacks amido- groups of a certain lability, e.g.,: C, HyNH,4+CH,0=C,H,. N= Chae Thus if labil amidogroups in enzyms would be changed in an analogous manner, the activity of this grouping would of course be lost, since the amidogroup as such has disappeared. The following table shows the more or less intense action of formaldehyde on enzyms. 1 Centralbl, f, Bakteriologie 12, p. 445. 2 Journal fir prakt, Chemie 1888, vol. 37. p. 104. Such observations were later on made also by various authors, Studies on the Lability of Enzyms. 59 Time in which idrovataene Formaldehyde.! Beene ia iatiod: Author. Diastasc 1% in 24 hours Bokorny. Myrosin 5% soon y Rennet 0.5% i lia Zymase 0.05 % a cele Sucrase 5% one hour at 54° C. Pottevin 7 Catalase 4% I hour Loew Pepsin 5% 24 hours fe Pepsin 4% 24 hours Sawamura Papain 0.4.% Soon at 40° Vines It is true that formaldehyde will act also. on hydroxylgroups and produce methylene-compounds as, e.g., with the polyvalent alcohols, yielding the so-called ‘formals,’ but in order to accomplish this, applica- tion of heat in presence of hydrochloric acid is required, hence the conditions are far different from those just mentioned. The inference that amidogroups of a certain lability are concerned in the activity of enzyms would receive further support, if it could be shown, that the enzyms take ‘ The commercial formalin or formal contains abaut 40% formaldehyde. 6c K. Aso. up free cyanogen and would lose thereby their activity.1 As to the different lability of amidogroups Loew expresses himself as follows: “ Die Amidogruppe kann unter gewissen Umstanden stabil, reactionsunfahig, unter anderen aber wieder ausserst labil und reaktionsfahig sein. Die Amidogruppe ist Z. B. im Urethan sehr stabil, im Harastoff labiler, noch mehr im Guanidin. Im Hydroxylamin und Diamid aber ist sie so energisch geworden, dass diese Stoffe selbst bei grosser Verdiinnung noch in alles lebende Protoplasma ohne Ausnahme eingreifen kénnen, d. h. Gifte allgemeinen Charakters sind. Folgende Formeln lassen die Einfliisse benachbarter Gruppen auf die Energie der Amidogruppe erkennen:”’ NEY NH, INES Ault as ie CO CO C=NH oleate \NH, \NH, Urethan Harnstoff Guanidin NH, NE; | NH, OH Diamid (Hydrazin) Hydroxylamin When dicyanogen acts on amines, it forms as chief compounds addition products with two molecules of amine. Thus 2(C,H,.NH,)+(CN),=C,H, -N=C—C=N-C,H,? Fo N Ws When, however, it acts on amidocompounds several products may result. Thus the chief product with amidobenzoic acid is cyancarbimidamidoben- zoic acid CN—C—NELC, H, COOEF iI NH 1 Also hydroxylgroups can take up cyanogen at the ordinary temperature as Zeew has shown with pyrogallol. But this is the only case known thus far. (Journ, f. prakt. Chem, Vol, XV, 326.) Very labil methylene groupings as in the acetylacetic ether also can react with cyanogen but only in presence of sodium ethylate C,1MjONa (W, Traube), ‘Thus far, however, only a few such cases have beer desc ribed, 2 This tormula has been shown by 7vemannto correspond better to the behavior of the cyananiline than the former imidoformula, Studies on the Lability of Enzyms. 61 It is noticeable, however, that not every amido compound and amide can combine with dicyanogen, and imidogroupings are not acted upon at all. Neither hydrazobenzol nor asparagine nor urea are acted upon. But Andreasch has shown that methylthiourea enters into reaction. The peculiar behavior of free cyanogen towards highly labil. amidogroupings have induced Loew and Tsukamoto! to test the behavior of a highly diluted aqueous solution of dicyanogen towards living organisms of the most different kind. These tests have revealed highly poisonous properties of dicyanogen rendering the presence of labile amido groupings in the proteins of living matter highly probable. It was now of interest to test whether it could also kill the enzyms. Since most enzyms belong in all probability to the protein group it could hardly be doubted that dicyanoger: would act on enzyms when applied in excess to a concentrated solution, since Loew has shown that cyanogen combines with ordinary albumin.? He applied solutions containing 10—25% albumin. In my experiments with enzyms, the dilution was a much higher one, since it was to be expected that very labile amido groupings would take up dicyanogen in high dilution, hence a reaction under this condition would admit a safer conclusion as to the degree of lability. Experiment with Pepsin. © grams of commercial pepsin? were dissolved in 200 c.c. water and divided into halves.4 Cyanogengas developed from 5 grams mercuric cyanid was passed through one of these bottles which, well closed, was left for 12 hours, Thereupon 10 c.c. of 2% hydrochloric acid were added and some fibrin, previously swollen in dilute hydrochloric acid, and kept for 24 hours at 28° C. The fibrin was dissolved rapidly in both cases 1 Forschungsberichte tiber Lebensmittel, Vol. I. No. 7;—Journ, College of Science, Tokyd, 1896. Cf, also These Bulletins Vol. II. No. 7. 2 Journ, f. prakt. Chem. 1877. % The solution of this sample was acid, + Some ether was added to thecontrol flask to prevent bacterial growth, 62 K. Aso. and neither the precipitation with nitric acid nor the saturation with ammonium sulfate did show any difference ; nor the colorimetric comparison of the biuret reaction. In a second experiment, 2g of pepsin were dissolved in 200 c.c. of water and 20 c.c. of a 2% hydrochloric acid added. One half served as control, while the other half was treated with the same quantity of cyanogengas as before, but in this case, the solution was kept at 35-40°C during the treatment. Moreover this liquid gvas placed in the incubator at 28°C for 20 hours before testing its proteolytic action. Equal quantities of fibrin and thin square slices of boiled egg white were now added to both liquids which were kept at 28°C for one hour. The fibrin was dissolved also here and the egg-albumin was almost wholly digested after 3 hours in both cases. In the third experi- ra) ment the conditions were again changed. Pepsin, 1g, was dissolved in 200 c.c. water and 1g of sodium carbonate (anhydrous) added. 100 C.c. of this solution were treated with the same quantity of cyanogen as before and left for 12 hours. After neutralizing 10 c.c. of 29 hydrochloric acid were added and fibrin. The result showed that the pepsin had been killed by the sodium carbonate. This result is not surprising considering the great sensitivness of pepsin toward alkaline liquids. Green reports that pepsin is injured by 0.002% soda solution after 1-2 hours at bodily temperature. Neverthless, a further experiment was made with the modification that the pepsin solution was rendered but very slightly alkaline with sodium carbonate. The passing of cyanogen from 5 grms of mercuric cyanide on heating took about one hour. Afterwards 10 c.c. of 2% hydrochloric acid were added to 100 c.c. of pepsin solution, the treated one, as well as the control. The same quantity of fibrin was added in both cases, but no digestion took place in either case. In the final experiment the acidity was hardly perceptible to litmuspaper; the further treatment was the same as just mentioned. In both cases, the fibrin was quite dissolved after one hour while the slices of boiled egg disappeared after 12 hours. 1 Langley and Lves found a distinetly inhibitory action to be manifested by the presence of as little as 0.0015 9% of sodium carbonate, - Studies on the Lability of Enzyms. 63 Experiment with Trypsin. I gram of commercial trypsin was dissolved in 200 c.c. of water containing 0.4g. sodium carbonate and divided into halves, one serving as control and the other being treated with cyanogen gas developed from 5 germs of mercuric cyanid. After 12 hours an equal quantity of fibrin and two thin square slices of boiled egg were put in each solution. After two hours at 28°C the fibrin was almost completely digested in both solutions, also the egg slices were very much attacked, but did not disappear completely after 20 hours. In the next experiment, the quantity of mercuric cyanid was doubled, but the result was the same as before. Experiment with Emulsin. A solution of 0.19% emulsin with 0.19 Na,CO, was treated with cyanogen gas developed from 5 grms mercuric cyanid. After standing for 48 hours, o.1g. of amygdalin was added to 10 c.c. of the solution. After one hour at 28°C, the peculiar odor of benzaldehyde was plainly perceptible like in the control case. The decomposition of amygdalin was also shown by the reaction with ammoniacal silver solution and with fuchsin solution decolorized by sulphurous acid, proving the formation of the decomposition products of amygdalin, viz. of glucose as well as of benzaldehyde. Experiment with Takadiastase. 0.2g¢. of commercial takadiastase were dissolved in 200 c.c. of water. This solution had a faint acid reaction and was rendered faintly alkaline by sodium carbonate. One half was treated with cyanogen developed from 5 grms mercuric cyanid while the other half served as control. After 24 hours standing the amylolytic action was compared. 10 c.c. of these solutions were mixed with 10 c.c. of 0.199 starch paste suspension 64 K. Aso. and kept at 30°C for 2 hours. On addition of iodine solution, no blue reaction set in, showing that the starch was transformed equally well in both cases.! On boiling with Fehling’s solution, a strong sugar reaction was obtained and upon warming with ammoniacal silver solution, a silver mirror appeared in both cases. The enzym had therefore not lost its activity by the treatment with cyanogen. Experiment with Oxidases. 45g. of a fresh radish root were triturated with addition of 100 c.c. water. Through 50 c.c. of this extract, cyanogen gas developed from 5 grms mercuric cyanid was passed while 50 c.c. served as control. After standing 15 hours, the cyanogen gas was replaced by air and the liquid tested for oxidizing enzyms in the usual manner, but the treated solution gave the color tests much weaker than the control. The experiment was repeated with the result, that when after 24 hours standing the color tests were made, they failed almost completely. Since dicyanogen in aqueous solution soon forms some prussic acid it was possible that some prussic acid had paralyzed the action of the oxidizing enzyms. Hence in the next trial the treated liquid was left to evaporate at 4o-50°C to remove the prussic acid, whereupon the guaiac and the cuaiacol test for peroxidase were readily obtained, only the guaiac test for oxidase was somewhat weaker. In all the cases here described adicyanogen has failed to destroy the activity of the enzyms, what reveals a great chemical difference between the lability of enzyms and the lability of the active proteins in the living protoplasm. Loew and Tsukamoto (Il. c.) have observed that a fresh solution of dicyanogen in water in a dilution of 1: 5000 kills bacteria and of 1 : 10000 1 Although this solution had a weak alkaline reaction, the diastase was not perceptively injured. Chittenden and also Grittzner have observed an injurious action of smal] quantities of alkalies, but Epstein and Schulze found that hereby this enzym is not destroyed, but merely “ paralysed,’ because by neutralization the activity is again restored, at least partially, Studies on the Lability of Enzyms. 65 phaenogams, algae and lower aquatic animals. Here exists then another striking difference in the chemical behavior of living protoplasm and enzyms. | The apparent indifference of enzyms to dicyanogen shows either that the amido groupings in the enzyms are not of sufficient lability or that they are protected by other neighboring atomic groupings, forming @) steric .obstacle in the molecule: The inference that there are no amido groups present at all would be improbable considering the behavior of enzyms towards formaldehyde. Further tests therefore seemed necessary to demonstrate the participation of the amidogroups in the activity of the enzyms. Here the behavior towards nitrous acid promised to furnish some clue. In my experiment with enzyms I added to highly diluted solution of sodium nitrate and sodium nitrite the theoretically necessary quantity of sulphuric acid.1 With those enzyms which are injured very easily by any kind of acids special care was necessary to apply nitric and nitrous acid in a very high dilution. The cause of the injurious action of nitric acid would of course be very different from that of nitrous acid. The former in high dilution would act like dilute sulphuric acid causing atomic migration of labil atoms while nitrous acid would act on labil amido groups in the following manner: NEeNO” ==/xjJ_N=N-—OH+H,0 and ie OH in the case of aliphatic compounds development of nitrogen would immediately follow the formation of a diazocompound: (x)—N=N—OH ——(x)—OH+N, —_—__—— - Diazocompound Experiment with Pepsin. 1). 0.3g of pepsin were dissolved in 300 c.c. water, and divided ito three equal parts. To one potassium wétrite and sulphuric acid in 1 In this connection it is also an interesting fact that a very high diluted nitrous acid (1 : 100000) is much more poisonous for lower organisms than nitric acid, as Zeew and Bekerny have shown, 66 K. Aso. high dilution were added in the calculated proportions to produce free nitrous acid of 0.19% in the liquid. To another potassium strate and sulphuric acid were added in such proportion as to produce 0:1% nitric acid, while the third portion served as control. The nitrous acid soon caused a yellowing of the pepsin solution, while nitric acid did not. After standing 24 hours,! an equal quantity of fibrin previously swollen in dilute hydrochloric acid and washed, further two square slices of boiled egg white and Io c.c. of a 2% hydrochloric acid were added. | After 1], hour at 40°C, After 1. houryat. Aoi. | | * Fibrin dissolved; egg | Egg white wholly Control. . ea! . white not yet. | dissolved. | Fibrin dissolved; egg 5 : ¢ re : | ; ete: Some egg white still Nitrous acid. | white almost unchang- ; remained. ed. Some pieces of fibrin dissolved; egg white Nitrous acid. gees | ee : beef ese attacked. | remained intact. | An equal quantity of fibrin was again added; After 4 hour, After 1 hour. After 17 hours, After 3 days. Contral Fibrin and egg a : white dissolved. Treated with Fibrin as well as Both almost wwe ten, nitric acid, egg white attacked. dissolved. All dissolved. Treated with Fibrin and egg Both Fibrin dissolved, | Egg white a little nitrous acid, white not dissolved. unchanged. but not egg white. attacked. i ae EE TL 2). In a second experiment 0.29% nitric acid and nitrous acid were applied in the same way as before. In the control solution however, 0.29 sulphuric acid was added to show the effect of the mere acidity. The solutions were kept at the ordinary temperature for 24 hours. ' The solution treated with nitrous acid developed a peculiar odor of certain nitro-compounds, | ; ie Studies on the Lability of Enzyms. 67 Hereupon 10 c.c. of a 1% hydrochloric acid were added to each solution further an equal quantity of fibrin and two square slices of boiled egg white. These solutions were kept at 40°C. After 2 days. - * - a ~ “ - After 4 hour. After 4 hour. After 1 hour. (at ordinary temp.) 0.2% sulphuric Fibrin dissolved fala almost dis- Fibrin dissolved. Egg white : : solved ; Egg white Egg white DES oe acid. very little. a } dissolved. unchanged. unchanged. 0.2% nitric nme ee eee ee eee | eee ved Egg white acid. dissolved. Boeee TE dissolved. unchanged, unchanged, 0.2% nitrous Not Some fibrin dis- Some fibrin and ; 2 cid Bircaived solved. Egg white —— egg white stil! ‘ [ not. unchanged. A further quantity of fibrin and egg white was added and kept at 40°C :: After $ hour. After 1 hour. After 20 hours. 0.2% sulphuric Fibrin almost dissolved, a ouoe BENGE i Ege¢ white dissolved - acid Egg white unchanged ge white hardly very much HS 88 Sees unchanged. 5 ; “Aor yA Ags : All fibrin dissolved 0 ; : Geo wee ee ee eS Egg white hardly ditto. : 88 as unchanged. 0,29% nitrous Much fibrin unattacked. | Much fibrin and all Egg | Egg white unchanged ; acid. Egg white unchanged. white unchanged. Some fibrin still present. 3). In’ the third test, o.1g. of nitrous, nitric and sulphuric acid were added respectively, each bottle holding 100 c.c. of 0.1% pepsin solution, and immediately heated at 35°C for one hour. 10 c.c. of 19% hydrochloric acid, further eggwhite and fibrin were then added. After 2 hours at 40° After 24 hours at 40° 0.19 sulphuric acid. 0.19% nitric acid. 3 + Fibrin dissolved. Egg white almost dissolved. | | ; 0.19% nitrous : Some fibrin. unattacked. acid. Ege white unchanged. 68 K. Aso. 0.1 germ. of nitrous, nitric and sulphuric acid was again added to these solutions respectively, and kept at 40° for one hour. An equal quantity of fibrin was then added. After 1 hour at 40° 0.2% sulphuric acid. All fibrin and all egg white dissolved. 0.29% nitric acid. -< + ” 0.2% nitrous acid. All fibrin and egg white undissolved. Experiment with Trypsin. Into three flasks holding 100 c.c. of a 0.5% trypsin solution, 0-05 grams of nitrous, nitric and sulphuric acid were added and kept at 40° for one hour. These solutions were now neutralized and to c.c. of a 2% sodium carbonate solution added and some fibrin. After 2 hours at 40° After 17 hours at 40° 0.05% sulphuric | Soc . | SG. ae Seeractn | Some fibrin dissolved, All fibrin dissolved. 0.05% nitric acid. | ”? ” ” ” ” ” 0.05% nitrous ae , om . a aa Fibrin undissolved. Fibrin undissolved, Laperiment with Emulsin. This test was made in the same way as in the last mentioned case. After 20 hours, 10 c.c. taken from each flask received o.1g. of amygdalin, After 15 minutes at 40°C, the following was observed. } Studies on the Lability of Enzyms. 69 Odor of benzaldehyde and prussic acid. Reduction with Control. ae ; : : 5 ; Fehling’s solution and with ammonical silver solution, No odor developed. No reduction took place with the 0.1% nitric acid. 1% above-named reagents. 0.19% nitrous acid, 33 9 ” The result did not differ when the amygdalin was added after neutralisa- tion of the liquids. 2). In the next experiment, the quantity of nitric and nitrous acids was reduced to 0.059§, while to the control solution sulphuric acid was added in the same concentration. After keeping for 16 hours at 18° these solutions were neutralized and o.5¢. amygdalin added. On keeping these mixtures now at 40° for three hours, the following was observed: The characteristic odor of benzaldehyd appeared . re ; 0.05% sulphuric acid. very plainly. 0.05% nitric acid. FA + 0.05% nitrous acid. No odor developed. These tests with pepsin, trypsin and emulsin show therefore, that nitrous acid destroys the activity of these enzyms more easily than do nitric and sulphuric acids in the same high dilution. In order to test for ketone groups, experiments were made with hydrazine, methylhydrazine and hydroxylamine. The solutions of the salts of these bases having an acid reaction were neutralised with sodium carbonate. From the amount of salt weighed out, the amount of the free bases was calculated. 1) Aydrasine. Pepsin: 100 cc. 0.19 pepsin solution + 19¢ free hydrazine. After two hours at 40° 0.29% hydrochloric acid and three flocculi of fibrin were added and kept again at 40°C. 70 kK. Aso. After 4 hour. After 2 hours. After 4 hours. 126 free Fibrin not attacked Not dissolved. Not attacked at all. hydrazine. at alle Fibrin wholly dissolved. Trypsin: 19% trypsin solution + 19 free hydrazine. After two hours at 40°C, 0.29% sodium carbonate and three flocculi /0 of fibrin were added, keeping the mixtures in the incubator. After 2 hours. After several days, 1% free hydrazine. Fibrin unattacked. Not attacked. Control, Almost dissolved. All dissolved. Diastase: Solution of 0.1% diastase + 1% free hydrazine. After keéping at 40°C for 2 hours some 0.1% starch paste was added and kept at 40°C, for 2 hours. After evaporating and removing the hydrazine with alcohol the residue was treated with water and tested with iodine dissolved in potassium iodid. 1% free hydrazine. | Blue starch reaction. Control. No starch reaction. Emulsin: o.£% emulsin solution + 19§ free hydrazine. After keeping at 4o°C for 4 hours, o.1 grm. amygdalin was added tO TO. cic; Studies on the Lability of Enzyms. 71 The odor of benzaldehyd and prussic acid % free hydrazine. ee nace developed very weak,* Control. The odor was very strong. These solutions were kept at 40°C and tested in the same way again: After 6 hours. After 8 hours, 1% free hydrazine. A very weak odor appeared. Trace of odor. Control. Very strong odor. Very strong odor, ee ee rr I have observed further that also zymase is easily killed by a 1% solution of hydrazine. 2) Methythydrazine. I. Experiment. Emulsin - : : 10.29 solution + 0.032% free methylhydrazine. Pepsin Trypsin 0.5% , " After keeping at 40°C for 1 hour was added: To emulsin: o.1g. of amygdalin Pebensin « G29 HCl ba ltypsin: —0.2%' Na,CO, Emulsin: After keeping at 40°C for 15 min. the enzym was still active but weakened somewhat by methylhydrazine. * It might be objected, that the diminution of the odor might have been due to the formation of benzyliden hydrazine but the control tests with the fresh mixture and that which had been tested after 8 hours digestion at 40° revealed a great difference in the intensity of the odor. The formation of benzyliden hydrazine from benzaldehyde and hydrazine in such high dilution does not take place instantaneously. 2 K, Aso. eee Pepsin Trypsin. a After 2 hours. Fibrin dissolved. Still some fibrin. Ae ais | 39 All dissolved, Il. Experiment. Emulsin 0.19% + free methylhydrazine 0.64% Pepsin 0.1% + - = After 2 hours at 4o°C was added: To Emulsin 0.2g. amygdalin, To Pepsin 0.29% HCl and 3 flocculi of fibrin. After 15 Min. at 40°, the tests showed that emulsin was still active in the 0.64% solution of free methylhydrazine, but it was weaker than in the control case. After one hour at 40°C, 0.64% free methylhydrazinc—Fibrin not dissolved at all, even not Pepsin after 2 days. Control : All fibrin dissolved. Il. Laperiment. Emulsin 0.19 + free methylhydrazine 0.64%. Kept at 32°C for 20 hours, and o.1 grm. of amygdalin added and warmed. Immediately tested: Control: —Odor of benzaldehyde. Treated :—No odor at all. fee — NI WwW Studies on the Lability of Enzyms, IV. Experiment. Takadiastase 0.1% + free methylhydrazine 0.32%. After 20 hours at 24° 10 c.c. of a 2% ‘starch paste were added, the mixture kept at 40° for 2 hours, then evaporated and extracted with alcohol to remove the methylhydrazine. The residue was treated with some water. Control: —Already after 4 hour, no longer any iodine reaction for starch. Treated :—Blue starch reaction. These tests leave no doubt that hydrazine and methylhydrazine injure enzyms very much or kill them. 3) Hydroxylamine. Pepsin 0.19% ++ free hydroxylamine 1°% free hydroxylamine 0.529 Trypsin 0.5% 4 : ae y» r: %» Diastase 0.19% + hydroxylamine 1% Emulsin 0.19% + > y After keeping at 40° for 2 hours was added: to pepsin: 0.29 HCl and three flocculi of fibrin. to trypsin: 0.2% Na,CO, and e : H to diastase: 0.1 grm. of starch in the form of starch paste. to Emulsin: 0.1 grm. amygdalin. and the mixtures kept at 40°C for 2 hours: Pepsin : Control :—Fibrin dissolved completely. 1% hydroxylamine :—Fibrin undissolved. Trypsin: Control :—Almost all fibrin dissolved. Fibrin unattacked. 0.5% hydroxylamine : 1% ” =e ” ” 1 The fibrin had dissolved already after 30 Min. 74 K. Aso, Even after 24 hours standing at the ordinary temperature, the fibrin was not dissolved where hydroxylamine had been added. Diastase : Control: —No starch reaction with iodine, 1% hydroxylamine :—Strong starch reaction.! Emulsin: immediately tested after adding amygdaline. Control :—Strong odor developed. 19% hydroxylamine :—Slight odor of prussic acid and benzaldehyde. This test was repeated by adding amyegdalin after heating the enzym solution with hydroxylamine to 40° for 4 hours. Control :—Strong odor. 1% hydroxylamine :—No odor. These observation on the injurious action of the hydroxylamine on enzyms are in accord with a former observation of O. Loew on diastase.? The following table shows the results obtained: l 1 Pepsin. Trypsin. Emulsin. Diastase. | Free | Kills at 0.29% in Kills at 0.059% in | Kills at 0.05% in | NO,H one hour at 40°C, | one hour at 40°C. 16 hours at “18°. Bree | Kills at 199 in two | Kills at 19¢ in two | Nearly kills at 0.19 | Kills at 196 in two iN Pls | hours at 40°C. hours at "40°C. in 8 hours at 40°C.} hours at 40°C. Free Kills at 0.649% in | Injures at 0.0329 | Kills at 0.649% in | Kills at 0.32% in NW H..CH,, two hours at 40°C. |in one hour at 40°C.| 20 hours at 32°C. | 20 hours at 24°C, Free Kills at 19% in 2 Kills at 196 in 2 Kills at 196 in 4 | Kills at 196 in two NH,.OH | hours at 40°C. hours at 40°C hours at 40°C. | hours at 40°. Conclusion. 1. Enzyms in high dilution are not killed by small quantities of dicyanogen. Hereby another essential difference between the chemical behavior of the living protoplasm and that of enzyms is established. 2 Small doses of iodine are changed to III by hydroxylamine, hence an excess must be added here to obtain the starch reaction. 2 Journ f. prakt. Chem, 1888. p. 104, Studies on the Lability of Enzyms. 75 2. Nitrous acid in very high dilution is more injurious for enzyms than cqually diluted nitric acid. 3. Hydrazine, methylhydrazine and hydroxylamine in dilute neutral solutions destroy the activity of enzyms. This would be best explained if the active grouping in the enzyms are cither aldehyde or ketone groups. According to Loew's present view, ketone groups alone can come here into consideration. Ueber fungicide Wirkungen von Pilzculturen, VON -Y. Kozai und O. Loew. Es ist seit lange bekannt, dass die Culturen mancher Bacterienarten Stoffe enthalten, welche das Wachstum anderer Bacterienarten hemmen oder verhindern. Bei den Culturen des Bac. pyocyancus ist es ein Enzym, welches geradezu manche andere Bacterien auflést.1 Auch die Ent- wicklung von Schimmelpilzen ist dfters auf Culturfliissigkeiten nicht méglich, in denen sich gewisse Bacterienarten entwickelt haben, obwohl es sich hier nicht um Aufloesung der Mycelfaden handelt. In neurer Zeit haben ferner L. Bourguelot und Heérissey? beobachtet, dass ein Extract von Aspergillus niger die Giartatigkeit der Hefe beeintrachtigt, die Hefe selbst aber nicht tétete. Diese Wirkung wurde selbst nach dem Autkochen der Loesung nicht aufgehoben. Die Beobachtung nun, dass der in Japan unter dem Namen Miso bekannte vegetabilische Kise selbst in der heissesten Zeit des Sommers nicht schimmelt, trotzdem er in hoch feuchtem Zustande dem Staube der Luft ansgesetzt in offnen Laden feil gehalten wird, bewog uns, auch dic Culturflissigkeit des Aspergillus oryzae auf fungicide Eigenschaften zu priifen. Jener Miso wird namlich mit Hilfe dieses Pilzes, resp. dessen Enzymen aus gekochten Soyabohnen dargestellt. Er enthalt 50-60% Wasser, 5-119 Kochsalz, 6-129 Rohprotein, 5-6.5% Fett und 13-249 Kohlehydrate und Extractstoffe. Die Reaction ist meist ganz schwach sauer. Der Kochsalzgehalt ist zu gering, als 1 Siehe R. Emmerich und O. Loew. Z. Hyg. 1898. 2 Jahresber, f. Thierchemie 1895, 5. 623. e) Y. Kozai und Q. Loew. NI A dass derselbe das Schimmeln verhindern kénnte. Wir liessen ihn in einem offnen Becherglase wahrend des Monats August bei einer alltaglich auf 33-35°C steigenden Temperatur stehen und beobachteten dabei eine allmalich sich entwickelnde Hefeschichte, welche dann von Sarcina iiberwuchert wurde. Schliesslich wurde auch diese durch Lac. prodigiosus verdrangt. Es wurde keine Spur von Penzcillium oder Aspergillus entwickelt. Die Reaction war schliesslich alkalisch geworden. Um nun zu prifen ob Aspergillus oryzae Stoffe produciren kann, welche auf ihn selbst sowohl als auf nahestehende andere Fadenpilze schadlich wirken, wurde jener Pilz auf folgender Loesung cultivirt : Pepton — 1% ; Zucker —0.5% K,H,PO, —0.2% MgSO, —0.02% Es wurden 4a Kolben, je 500 cc. dieser Loesung enthaltend, auf- gestellt und nach dem Sterilisiren inficirt, am 18 September. Um die Sporenbildung zu verhindern und die Mycelbildung zu begiinstigen, wur- den die Kolben taglich umgeschittelt. Einer der Kolben wurde am 6 October gedffnet, der Inhalt (der noch etwas unzersetztes Pepton enthielt) durch sterilisirte Filter in sterile Kolben filtrirt und ein Teil direct, der andere nach dem Aufkochen mit Penicilliumsporen inficirt. Nach einigen Tagen zeigte sich in beiden Flaschen eine sehr langsam sich entwickelnde Schimmelvegetation. Es wurden desshalb die anderen Kolben noch langer stehen gelassen, bis das Mycel den Nahrboden erschépft hatte und dem sAbsterben unterlag. Das letztre schien uns aus der allmialich ein- tretenden Schwarzfarbung der Fliissigkeit (Folge von austretenden oxy- direnden Enzymen?) zu folgen. Am 20 November wurden nun zu einem Kolben 2¢ sterilisirtes Pepton gesetzt und dann wie oben verfahren. Es ergab sich, dass diesmal selbst nach 11 Tagen bei 10-16°C. keine Spur einer Schimmel- vegetation eintrat. Auffallig war, dass auch in der einen Moment auf- geckochten Portion sich keine Entwicklung nach der Impfung zeigte. Its muss also die Production eines gewissen fungiciden Stoffes durch Ueber fungicide Wirkungen von Pitzculturen. 79 den Pilz Aspergillus oryzae gefolgert werden. Diese Substanz ist aber fiir verschiedene Pilze nicht in gleichem Maase schiadlich. Im Anhang hiezu wurde noch ein zweiter Versuch mit Pezzccllium sporen gemacht, welche diesesmal auf eine nur schwach alkalisch rea- girende Culturfliissigkeit des Bac. pyocyaneus ausgesit wurden. Es fand auch nach Wochen keine Entwicklung von Penicillium statt, wahrend im Controlversuch dieselbe sehr lebhaft war. Zur Frage der Existenz des Pyocyanolysins. VON O. Loew und Y. Kozai. Bulloch und Hunter hatten vor mehreren Jahren beobachtet, dass . die Culturen des Bac. pyocyaneus einen Kérper enthalten, welcher Blut- kérperchen auflést. Sie fanden ferner, dass dieser Korper vorzugsweise in den Zellen bleibt, so dass Filtrate der Culturen weit schwacher wir- is ken, als sterilisirte unfiltrirte Culturen. Darauf hin haben wir gesucht, Bedingungen zu finden unter welchen dieser Korper, den Balloch und Hunter far ein Enzym hielten und Pyocyanolysin nannten, in besonderem Maase entsteht.2 Wir beobachteten dabei, dass eine Peptoncultur bei reichlichem Luftzutritt die Eigenschaft Blutkorperchen zu lésen besonders stark zeigte, aber fiir Mause ganz harmlos war, wahrend die Bouillon- cultur bei nur geringem Luftzutritt toxische Eigenschaften hatte und in weit geringerem Maase Blut léste. Es war fiir uns allerdings etwas iiberraschend, dass ein Blut lésendes Enzym bei subcutaner Injection harmlos fiir Miiuse sein sollte. Indessen die Beobachtung von Bullock und Hunter waren auch von Weingeroff,® ferner von Nencki und Szeder gemacht worden, Da ferner B. pyocyaneus besonders reichlich enzym- bildend ist’ war auch die Bildung eines blutlésenden Enzyms_ nicht 1 Centralbl. f. Bakt. Band 28, S. 866. Diese Bulletins, Bd. 4, No. 5. Centralbl. f. Bakt. Bd. 29, S. 777. tw i] 4 Briefliche Mitteilung von M. Nencki an den einen von uns. 5 In neurer Zeit constatirte Zi7smann (C. Bakt. Bd, 29, S. 848) die Bildung von Lipase, ferner die eines elastische Fasern lésenden Enzyms (Ibid. Bd. 33, S. 2), durch den Bae, pyocyaneus, Nach Eizykmann erkennt man am einfachsten die blutlésenden Eigenschaften mit Blut-Agar. C. Bakt. 29. g g 9 S. 847. 82 0. Loew und Y. Kozai. unwahrscheinlich, da ferner unsere Culturen nur schwach alkalisch reagir- ten, suchten wir den Grund der Blutloesung auch nicht im Alkaligehalt, um so weinger als nach JZyers! hinreichend schwaches Ammoniak keine Haemolyse verursacht, wenn die Blutkérperchen in physiologischer Koch- salzloesung suspendirt sind.2,) Immerhin war es uns auffallend, dass die haemolytische Wirkung durch Kochen der Loesung nicht aufgehoben wurde und sich keine giftige Wirkung bei Mausen constatiren liess. In neuester Zeit hat nun Fordan*® ebenfalls die haemolytische Wir- kung von Pyocyaneuskulturen beobachtet, aber dieselbe lediglich als Folge des Alkaligehalts der Culturen erklart. Werden die Culturen genau neutralisirt, so bleibt die Haemolyse aus. Wir haben diesen Ver- such wiederholt und kénnen diese Beobachtung im Wesentlichen be- statigen. Als 2 cc. einer 59 Aufschwemmung von Blutkérperchen mit 2 cc. der 15 Minuten auf 60° erwairmten Cultur 24 Stunden im Brutschrank blieben, war Lésung der Blutkérperchen eingetreten; als aber die Cultur vorher genau neutralisirt wurde, war nach 24 Stunden keine Haemolyse cingetreten. Erst nach einen weiteren Tag trat allmilich Loesung ein, doch kann dieses kaum auf cine Enzymwirkung gedeutet werden. Weitere Versuche haben ergeben, dass schon auffallend geringe Mengen von Natriumcarbonat Haemolyse herbeifiihren kénnen. Als 2 cc. einer Blutkérperchenaufschwemmung mit 2 cc. einer 0.0019§ Sodaloesung 24 Stunden bei 32° gehalten wurden, war schon etwa die Hialfte, nach 20 Stunden alles gelést. Es ist dieses um so auffallender, als das Blut im lebenden Thier doch auch eine alkalische Reaction besitzt. LC, Bakt. 25) 9.2975 2 Erst wenn eine gewisse Menge secundires Natriumphosphat zugeftigt wird, erfolgt Haemo- * C, Bakt., Bd. 33, 5. 274. ae Op On the Microbes of the Vukamiso. BY S. Sawamura. The name of Wukamitso is given in Japan to a preparation, resulting from the spontaneous fermentation of a mixture of rice bran, common salt and water. It is used for softening and rendering palatable certain fruits and roots. According to /zouye} it has following composition. Ry aiens cec. eee Seca she c's a 75.6% WA CUIG RACs (acencr cee es « : 2.6 Sua hs. 5 Soe, Beek he eee “A ee Hodim enlorid......--.. i MEN 8.1 Proteids, amidocompounds, fats, : 10,4 mineral matters, starch. The most striking chemical change caused by that fermentation is the increased production of sugar and acids. The writer estimated the quantity of acides and sugar in Mukamiso fermented by keeping the fresh mixture in a warm (20—25°C) place for 20 hours.2 The quantity of normal soda solution necessary for neutralizing 1oocc of the filtrate was 5.2 cc, while the acidity of the original rice bran mixture was only o.4cc. The quantity of sugar calculated as dextrose was as follows :— Ciicinal mixtures sstise....:.... 0.04.1. 96 TLEET URES Dla eckson ots » “Creu GEA.) By preparing a plate-culture of chalk-glucose medium with freshly pre- pared Wukamiso four kinds of bacilli which produced acids were isolated. The microbes have the following properties. 1 This bulletin Vol. IT. No. 4. 2 The mixture consisted of roo gr of rice bran, 50 gr of common salt and 1 litre of water. $4 S. Sawamura: INO tr: Form. The cell cultured in bouillon for 24 hours at 37°C is 0,5 wide and 1—2y long. Two are generally united ; they are motile by peritric flagella. Spore formation is not observed. Staining. Gram’s method negative. Oxygen. Aerobic. Bouillon, A feeble scum and a little deposit are formed. Gelatin plate-culture. A round, light, yellow, moist, bright, sharply defined colony. An elevated point is observed in the centre. It grows to a moderate size. Deep colony appears as a white point. Gelatin streak-culture. A light yellow, moist, homogeneous colony. Gelatin is not liquefied. Gelatin stab-culture. Thread-like growth to the bottom. Agar streak-culture. A white, homogeneous colony, condensed water clear. Potato culture. An elevated, moist, at first gray but afterwards yellow colony. Milk culture. It is coagulated with an acid reaction. Gas. It is evolved in glucose-bouillon. The gas consists of CO2 and H. Indol reaction. Positive. Chemical activity. It does not saccharify starch. Acid produced ina mixture of 10 gr of rice bran and 100cc of water for 3 days at 36°C was 0.780% calculated as lactic acid from the quantity of normal soda VA) solution necessary for neutralisation. Nian 22; orm. The cell is 0.64 wide and 2—4y long, and has rounded ends. It is motile and flagella seem to grow in one end of the rod. Gram’s method. Positive. Oxygen. Aérobic. Bouillon. A feeble scum and a moderate deposit are produced, but in elucose-pepton water a thick scum which finally breaks. Gelatin plate-culture. A round, sharply defined, somewhat transparent, On the Microbes of Nukamiso. 85 homogeneous, moist colony, which never grows larger than 2 mm in diameter. Gelatin streak-culture. A light yellow, moist, homogeneous colony. Gelatin is not liquefied. Gelatin stab-culture. Thread-like growth to the bottom. Agar plate-culture. A round, elevated, somewhat transparent, moist colony which does not grow larger. By weak magnification it is the same. Deep colony is a white point. Agar streak-culture. A yellowish white, moist, bright, homogeneous colony. Potato culture. A dark yellow, moist, bright, homogeneous colony, Milk culture. It is coagulated with an acid reaction. Gas. It is not evolved in glucose-bouillon. Indol reaction. Negative. Chemical activity. Starch is not saccharified, and sugar is not formed also in Wukamzso by this bacillus. Acid produced in a mixture of 10 gr of bran and 100cc of water for 3 days at 36°C was 1.327% calculated as lactic acid, and that produced in glucose-buillon con- taining some Ca CO3 in 3 days at 36°C was 1.007% of lactic acid calculated from CaO dissolved. The acid produced was found to be lactic acid by examining the properties of the zinc salt.! Some alcohol is formed from glucose, which was confirmed by the formation of iodoform. Since the already known lactic ferments such as Bacillus acidi lactici Hueppe, Bacterium acidi lactici Grotenfeldt and Kozaz’s bacilli are all not motile this microbe seems to be a new species. No. 3. Form. The cellis 0.4 wide and ty long, and it is motile by peritric flagella. 1 The nutritive solution of glucose with CaCQ3, in which this bacillus was cultured, was filtered. The filtrate, after having been acidified with P205, was evaporated to dryness, ‘The residue was treated with either and filtered. The filtrate was evaporated but no crystal was formed, the residue being a syrupy mass. It was neutralised with Zn CO3, and by examining the crystal-form of the zine salt and its behavior towards alcoholic ammonia, it was proved to be zinc lactate. 86 Ss. Sawamura: Two are usually united, and spore-formation is not observed. Gram’s method. Negative. Oxygen. Aérobic. Bouillon. It becomes turbid, but no scum is formed. Gelatin plate-culture. A round, elevated, sharply defined, white, moist, bright, homogeneous colony. By weak magnification it is the same. Deep colony is a white point. Gelatin streak-culture. A white, moist, bright, homogeneous colony. Gelatin is not liquefied. Gelatin stab-culture. Thread-like growth to the bottom. Agar streak-culture. A white, moist, bright, flat colony. Milk culture. It is coagulated with an acid reaction. Potato culture. A gray moist, elevated colony. Gas bubbles are formed on the colony. Gas. Gas consisting of CO2 and H is vigorously produced in glucose bouillon. Indol reaction. Negative. Chemical activity. It does not saccharify starch. Acid produced in Nukamiso above described for 3 days at 36°C was 0.654% calculated as lactic acid and that produced in glucose bouillon containing CaCO3 in 3 days at 36°C was 0.453% of lactic acid calculated from dissolved CaO. This microbe can produce the characteristic smell of Wakamiso. This is probably Bacillus chologenes Avase, which resembles very much the coli-bacillus. No. 4. Form. The cell is 0.5—0.74 wide and 2—4y long and is motile by peritric flagella. Spore-formation is not observed, Gram’s method, positive. Oxygen. Aérobic. Bouillon. A stick scum is formed, the medium remaining quite clear. Gelatin plate-culture. A round, flat, gray, roughly defined colony which erows very large. Gelatin streak-culture. It liquefies quickly gelatin. On the Microbes of Nukamiso. 87 Gelatin stab-culture. Thread-like growth to the bottom, the surface being quickly liquefied. Agar streak-culture. A light brown, folded, characteristic colony. Milk culture. It is coagulated with an acid reaction. Potato culture. Characteristic folded colony which is at first faintly red, and changes to gray afterwards. Gas. It is not evolved. Indol reaction. Positive. Chemical activity. It saccharifies starch, and produces 0.296% of glucose in a mixture of 10 gr of bran and toocc of water for 24 hours at 36°C, and acid produced in the same mixture for 3 days at 36°C was 0.770% calculated as lactic acid. A feeble red tint is produced in Nukamiso. By these properties this microbe is regarded as Bacillus mesentericus ruber Glodig. From an old Nakamiso the writer isolated a bacillus which was identified to be Bacillus mesentericus vulgatus Fliigge and a kind of Kahm- yeast, which produces a weak alcoholic fermentation on a nutritive solution containing glucose. In order to see which microbe produces most acid they were inoculated into a sterilized Nukamiso and kept for 24 hours at 36°C. The quantity of normal soda necessary to neutralize 1oocc of the filtrate from the above culture was as follows :— Pet oriiwitn) 2 Oo ees etet 7.9 f.ocu)!- L2ce miter asi Roe ctu ieee geo Nes! ! x.y (205, EMSS aN USEI. wth Phvwte Used “Tjca) IO,;; ICRA MERON Sas COME ks, wax, IZ yy No. 2.+No. 4. kas Moke 5 No. 1.+No. 2.+No. 3.4+No. 4. 25.0,, From these figures it is clear that the chief acid producer is Bacillus No. 2, but the symbiosis with the other microbes increases the production of acid. The smell characteristic for Wukamiso is probably produced by a Mesentericus species, and the organism No. 3. The production of sugar in Nukamiso is solely due to the activity of the Mesentericus species. 88 S. Sawamura: The species of saccharomyces present does not participate in the fermentation, since its fermentative power is nearly nil. The writer observed in the case of saccharification of mannan by Bac. mes. vulgatus, the acceralating action of a certain wild yeast on this bacillus.!_ In order to see whether the function of this wild yeast be analogous to it the mixture of Bac. mes. vulgatus with the saccharomyces was cultured for 3 days at 20°C in glucose-bouillon containing CaCO3 and the quantity of CaO dissolved by the acid formed was determined. Control. btw.) saw lo eaatsedtones Hes) aera Bac. mes. vulgatus+the Saccharomyces. 0.910,, In the second experiment a mixed culture of Bac. mes. vulgatus and that yeast were left to act upon starch, 29§ of which were suspended in bouillon. The sugar formed in one week at the ordinary temperature was as follows :— Control! sg. 65. i ee ae Bac. mes. vulgatus 4-the wild yeast. ... 0.82,, From these figures it follows that the saccharomyces has indeed some accelerating effect upon the action of the bacteria. We may conclude as follows :— I. By the fermentation of Vwkamiso sugar and acids are formed in a moderate quantity. II. In fermented Mukamiso there are present various microbes, of which the writer isolated four kinds of bacilli and a Saccharomyces. III Sugar is produced exclusively by a Mesentericus species. IV. Several microbes that can produce acid are present in Vukamiso, but the chief acid-producer is a bacillus, which seems to be a new species. V. The aroma characteristic for Mukamiso seems chiefly to be pro- duced by a Mesentericus species. VI. The Saccharomyces present in Wuhkamiso seems to have no other effect than to acceralate somewhat the bacterial actions. 1 This bulletin Vol. V No 2, —_—~o1co—_——_ - =". ~~ Ueber den Kalkgehalt verschiedener tierischer Organe. VON M. Toyonaga. Das Muskelfleisch hat nach mehreren Autoren einen hdheren Gehalt an Magnesia als an Kalk, was wahrscheinlich mit der relativ geringen Zellkernmasse zusammenhingt.! Es zeigt sich jedoch ein Unterschied zwischen dem Kalkgehalt der Muskeln von Batrachiern und Fischen einerseits, und demjenigen der Muskeln der Warmbliiter anderseits; so ergiebt sich im Durchschnitt fiir tooo Teile frischen Muskels von Warm- bliitern 0,0954 Teile Calcium, bei Kaltbliitern aber 0,2913 Teile Calcium. : eee Ca re Ferner ist das Verhiiltniss Me der Muskelgewebe beider Tier- SRS) eruppen sehr verschieden. Muskel der Kaltbliiter, Muskel der Warmbliiter.2 Ca “Mge_ 1.26 0.34 Von einigem Interesse schien es nun, die glatten mit den quergestreiften Muskeln in dieser Hinsicht zu vergleichen, da die glatten Muskeln eine niedere Entwickelungsstufe des Muskelgewebes darstellen, und die relative Grésse der Zellkerne verschieden ist. Wergleichende chemische Unter- suchungen beider Muskelarten sind nur spirlich vorhanden. 4 Vergleiche meine fritheren Abhandlungen in diesen Bulletins, Band 5. 2 Durchschnitt aus mehreren Bestimmungen von Muskeln verschiedener Tiere nach A’ass, > gO M. Toyonaga. Von Interesse ist die Beobachtung Vincents,! dass die glatten Muskeln 6-8 mal so viel Nucleoproteid enthalten als die quergestreiften, und dass der Herzmuskel einen Uebergang zwischen beiden bildet. Beide Muskel- arten geben ein Salzplasma, welches entweder von selbst gerinnt oder durch Verdiinnung. Ob das Mehr von Nucleoproteid in den glatten Muskeln ginzlich dem relativ grésserem Zellkern der glatten Muskelfasern zuzuschreiben ist, wire noch zu untersuchen. Immerhin schien es von se yr Cae 2 Interesse, das Verhiltniss Ve 2 beiden Muskelarten von demselben LV (Oy g Tiere zu bestimmen. Ich wihlte die Schenkelmuskeln des Pferdes und verglich sie mit den Bauchmuskeln. Leider stellen die letzteren Muskeln keineswegs nur ein Gewebe aus glatten Muskelfasern dar, sondern enthalten noch quergestreifte Muskelfasern und Bindegewebe. Es kénnen daher die erhaltenen Zahlen nur annahernde Werte fiir die glatten Muskelfasern sein. Ich verfuhr im Wesentlichen nach der in meinen friheren Arbeiten (1. c.) erwahnten Methode und erhielt die folgenden Resultate. In 1000 Teilen frischer Substanz sind enthalten : Ca ‘aO MeO Ca Ig Mg ap ; 0.2 (uergestreifter Muskel des Pferdes, 0,064 0,322 = * , : oO. Glatter Muskel des Pferdes, 0,07 0,292 = Man erkennt hieraus, dass in der Tat die glatten Muskeln, obgleich noch gemischt mit quergestreiften, cinen etwas héheren Kalkgehalt ergeben als die quergestreiften. 1 Zeitschrift f. physiolog, Chemie. Band 34, 5, 417, (1902). ——— Ueber den Kalkgehalt derschiedener thierischer Organe. gI Hodensubstanz. Obgleich die Hoden driisige Organe sind, so erscheint doch die Zellkernmasse derselben geringer als die der Leber oder Pancreasdriise. Kalk- und Magnesiagehalt der Driisen scheint manchmal, vielleicht unter pathologischen Einfliissen, abnormen Schwankungen zu unterliegen, jedoch nicht in dem Sinne, dass der Magnesia-gehalt tiber den Kalkgehalt steigt, sondern dass der Kalkgehalt enorm zunimmt und der Magnesiagehalt abnimmt. So fand, Zimzmg1in der Asche der Pancreasdriisen von zwei krebskranken Frauen das eine Mal (a) Ca=2,55% und Mg=1,48%, das andere Mal (b) Ca=16,94% und Mg=0,37%.? Die Aschenprocente der frischen Driisen waren nahezu gleich, namlich 1,04 und 1,02 resp. Auf 1000 Teile frischer Substanz berechnet wiirde sich ergeben : (a) (b) Ca—o.2663 1.7356 Mg—o.1541 0.0383 Dieses abnorme Sinken des Magnesiumgehalts im Falle (b) beim Pancreas, erinnert an einen ganz ahnlichen Fall, beobachtet an der Leber, von Ordtmann. Dieser’ fand (1858) in der Leber 1.19% Asche und in dieser Asche= 3.62% CaO und nur 0.19% MgO, oder 0.289 Teile Ca und 0.017 Teile Mg auf 1000 Teile frischer Substanz ; es ist also in diesen abnormen Fallen: Pancreas (b). Leber (nach Oidtmann). Me > 45-3 17.0 1 Die anorganischen Bestandteile des Pancreas, Wiirzburg 1899. 2 Die eine Frau (a) litt an Magenkrebs, die andere (b) an Eierstockkrebs, 2 In der Asche der Milz fand dieser Autor auf 7,489°§ Kalk nur 0.49% Magnesia ; es ist also hier , CG das Verh, We =18.1 Q2 M. Toyonaga. Sonst bewegt bei Driisen sich dieses Verhiltniss ‘i zwischen 1 und 6. Aus Hoden von Fischen wurden bekanntlich interessante Substanzen gewonnen, die Protamine, aber es existirt doch noch keine vollstaindige quantitative Analyse des Hodens von Saugetieren,! woraus wir die Menge Albumim, Globulin, Nucleoprotein, Mucin, Lecithin, etc. entnehmen kénnten. Sogar die Trockensubstanz ist nicht genau bestimmt worden. Mitescher giebt zwar an, dass der Wassergehalt 75 procent und die Trocken- substanz 25 procent betrage; wahrscheinlich hatte er aber den Hoden gemischt mit Bindegewebe der Bestimmung unterworfen. Bei meiner Bestimmung entfernte ich das Bindegewebe so sorgfailtig wie méglich und fand dann den Wassergehalt weit bedeutender, naimlich 85.39%. Ich analysirte die Hodensubstanz des Pferdes und des Stieres? mit folgendem Resultat : Tn 1000 Teilen. Ca Total Asche, CaO MgO Mg Pferdehoden 9.550 0.096 0.256 os Stierhoden a) 9.943 0.102 0.214 O.51 I b) 10.109 0.091 0.237 ' Vor Kurzem hat Zevene aus Rinderhoden eine Nucleinséure dargestellt, welche bei Spaltung unter andern Guanin, Thymin und Cytosin lieferte. ? Den Chlorgehalt der Stierhodenasche fand ich zu 0.4189. Bei der Pancreasasche betrigt derselbe 2.5-2.6%. Ueber den Kalkgehalt derschiedener thierischer Organe. 93 Beim Vergleich der Trockensubstanzen ergiebt sich : | Quergestreifter Milchdriise Hoden Muskel des Pferdes. des Rindes, des Pferdes, CaO 0.0323% 0.2517% 0.668% MgO 0.1619 % 0.0639 % 0.1773 7% Ca “Mg 0.24 4.67 0.451 Wir finden daher, dass der Kalkgehalt des Hodens geringer ist als der von Pancreas, Milz und Leber, dass aber andererseits das Verhialtniss Ga Moc D ein weiteres ist als bei den Muskeln der Warmbliiter. Das Secret des Hodens ist aber sehr kalkreich, in Uebereinstimmung mit dem anschn- lichen Gehalt an Zellkernen (Spermatozoen). Es enthalt nach Z. Slowzoff? frisches Menschensperma=o0,90% Asche, ferner 9.689§ Trockensubstanz und 0.199% Nuclein (29§ des trocknen Spermas). Der Kalkgehalt in der Asche von zwei Alkoholfraktionen des Spermas betrug 22,409¢ und 15,08 %. Analytische Belege. Frische CaCO, | Mg,?.0, Ss ie sd Wasser. Ashe. (inder =| = (in der Substanz. Hiilfte). Halfte). Quergestreifter Muskel. 233.6916 g. 187.243 £. 2.032 &. 0.0134. 0.1045 s. Glatter Muskel. 231.0862 ,, 187.1316 ,, 2:1236.,, 0.0147 ,, 0.0938 ,, Ilodes des Pferdes. 120.6913 ,, 103.3148 ,, ISES21); 0.0103 ,, 0.0427 ,, Iloden des Stiers (a) 201.5540 ,, LPR 55 2. . 0.0184 ,, 0.0609 ,, (b) 186.8451 ,, 161.9256 ,, 1.9196 5, 0.0152 ,, 0.0612 ,, 1 Das Verhaltniss Ca_. , :; Meg in der Lunge niihert sich mehr dem in den Hoden als dem in der Leber und Milchdriise. Schmidt fand in 1ooo Teilen Lunge (Mensch) 1.9 CaO und 1.9 MgO, also Ca 1.18 Mg I 2 Zeitschrift f, physiolog. Chemie, 35, p. 358. 94 M. Toyonaga, Behufs ciner Uebersicht iiber die bisher in Bezug auf den Kalkgehalt tierischer Organe erhaltenen Resultate lasse ich folgende Zusammenstellung folgen. In 1000 Teilen frischer Substanz sind enthalten Calcium : Muskel des Warmbliiter 0.057 (Bunge, Mittel). ys “ s 0.095 (Katz [1896], Mittel). Ouergestreifter Muskel (Bferd) 0.046 (Toyonaga | 1902] ). Glatter Muskel (Pferd) 0.050 5 Weisse Hirnsubstanz (Kalb) 0.041 ” A 24 (Pferd) 0.037 5 Graue Hirnsubstanz (Kalb) 0.263 5 . 3 (Pferd) 0.778 a Milchdritise (Kuh) 0.600 * Hioden (Pferd) 0.069 r (Scien) 0.069 n Leber (Mensch) 0.284 (Oidtmann). Periphere Nerven (Pferd) 0.568 (Toyonaga). Es zeigt sich ferner, dass mit Zunahme des Kalks in driisigen Organen die Zunahme der Magnesia keineswegs gleichen Schritt halt, sondern in manchen Fallen sogar sehr gering bleibt. Vergleichen wir das Verhaltniss Ca/Mg beim Muskel und der weissen Hirnsubstanz mit dem in drisigen Organen und der grauen Hirnsubstanz, so ergiebt sich: Ca/Mg. Muskel der Warmbliiter 0.34 (Katz, Mittel). QOuergestreifter Muskel (Pferd) 0.24 (Toyonaga). Glatter Muskel (Pferd) 0.29 “4 Weisse Hirnsubstanz (Pferd) 0.30 9 k * (Kalb) 1.14 Graue Hirnsubstanz (Pferd) 2.80 » 7 a (Kalb) Lye Y Milchdriise (Rind) 4.69 " Niere 1.84 (Aloy). » (Rind) 2.98 (Gossmann). Ueber den Kalkgehalt derschiedener thierischer Organe. Niere (Mensch) Milz (Rind) Milzpulpa (Rind) Bindegewebe der Milz (Rind) Pankreas (Mensch) Lunge . (Pferd) Hoden (Stier) ‘i (Pferd) Periphere Nerven (Pferd) Ca/Mg. 4.25 (Gossmann). 2.52 (Ribaut, Mittel). 2.79 (Aloy). 2.70 (Ribaut). 3-45 ” 1.73 (Liining 1900). 4.75 (Gossmann). 1.20 (Schmidt). 1.36 (Toyonaga). 0.51 . 0.45 - 1.56 “ | | | ) | On the Influence of Different Ratios of Lime to Magnesia on the Growth of Rice. A number of experiments to find the most favorable ratio of lime to magnesia for plant-growth have been made at this Institute, under Prof. Loew. Furuta studied in this regard the behavior of buckwheat, oats and cabbage in soil culture and the writer! that of barley, soy-bean and onion in water culture, and of the mulberry-tree in water as well as in soil culture. Recently, Katayama? has also carried out several sand and soil cultures with onion, oats and buckwheat on this line. In all those cases, it became evident that the maximum yield depended, other things being equal, upon a distinct ratio of lime to magnesia and that the best ratio is not the same with every kind of crops. Since rice-culture is a most important factor® in agriculture of Japan, it occurred to me that this principle should be also studied with the rice crop. The soils differ widely in chemical composition and the farmer never knows whether liming would be in order or not. Generally, however, the farmers of Japan apply too much lime and the injuries thus produced have induced the local government of Kiushiu to issue a law prohibiting the use of lime. Besides the depression of the harvest also a greater brittleness of straw and grains and a relative decrease of protein result from excessive liming. The content of lime and magnesia in the straw and grains of rice plants are the following in an average : 1 Bull, College of Agric, Tokyo, Vol. IV. No. § and Vol, No, 4. 2 This Bulletin, p, 102. 3 The annual production of rice in Japan (without Formosa) amounts in average to 41 millions Koku(=7.4 Mill. Liter.) while the importation amounts to a value of at least 5 million yen annually, 4 Bull. College of Agric, Toky6, Vol. T. No. o. 98 K. Aso. In 100 parts of.air dry matter : CaO MgO CaO: MgO _ (straw 0.26 O.1 EAScot Paddy rice} ; : : ; not whitened grains 0.03 0.09 Tecee straw 0.31 0.24 Lacan Upland rice Pp , : not whitened grains 0.02 0.07 12) 26235 It will be seen from these figures, that the ratio of lime to magnesia in rice is smaller than that in many other plants. My experiment was carried out as follows :— Seven Wagzuer’s porcelain pots were filled with 7 kilo of airdry sifted soil taken from a paddy field which had not been cultivated for several years. The quantity of available lime and magnesia in this soil was deter- mined by extracting the soil with cold 1o % hydrochloric acid for 48 hours with the following result : In 100 parts of dry soil ; CaO 0.70 MgO 0.60 The ratio of lime magnesia was now changed in six pots by mixing the soil with calcium carbonate or pure magnesite! (finely powdered) to reach the following ratios : — Quantity of Quantity of Pots. Calcium carbonate Magnesite CaO : MgO added. added, gr. gr. a 166.24 fe) bores b 122.86 fe) rete € 79.46 fe) Saat d 36.07 fe) 2 tcvat e (original) fo) fe) I : I (nearly) f o) 68.3 ee g ) 127.4 rsa 1 This mineral was imported from Germany and contained only minute quantities of lime. On the Influence of Different Ratios of Lime to Magnesia on the Growth of Rice. 99 As general manure for each pot served : Ammonium sulphate 15 grm. Sodium phosphate I5 grm. Potassium carbonate ! 1o grm. On July 13, the young rice plants? (about 36 cm. long) were transplanted from the seed bed. Each pot received three bundles. One bundle was made up of three individuals of equal size. Although this experiment was carried out in a glass house, the treatment was the same as in the field. Towards the end of August the difference in plant growth became very marked, the plants in e showed the best growth of all. These plants also flowered first (Sept. 9.). On September 18, all plants were in flower and on that day a photograph was taken which is reproduced on plate X and which exhibits the difference in development very well. It might be surmised that some ammonia of the sulfate was transformed into carbonate by the potassium or calcium carbonate added and volatilized; and this loss of some nitrogen might have something to do with the difference in growth. But it must be remembered that not only was the dose of nitrogen a very heavy one (ratio: 600 kilo N per ha), and that much more nitrogen was present than could be possibly utilized, but also that the soil was so rich in humus (11%) that a considerable absorption of ammonia was undisputable. Neverthless further experiments are contemplated with such a modification that even a small loss of nitrogen with be practically excluded,? viz. P,O, will be applied as superphosphate and K,O as sulfate. On November 6, the crop was harvested and left to become airdry. The weight was as follows: 1 This was applied separately, later. 2 The variety name was Satsuma. 3 P. Wagner has calculated from many trials with oats, that in average 1 gram of ammonica nitrogen can yield 56.97g. straw + 41.33g. of grains, while r gram of nitrate nitrogen, 57.03g. straw + 42.53g. of grains, From these data it may be seen, that there was a considerable excess of . nitrogen in my manured soil compared with the harvest of rice, which in all probability would show not a very different behavior from the related oats. 100 K. Aso. ee eer ee nr en. TTT —=$$—$————————— Pots. — Full grains. Empty grains. Straw, Total. 5 gv. Sr &/ ST. a | = 20.5 2.0 53-5 76.9 | 4 b =) 30.5 1.5 59.5 91.5 c ae 44.0 2.0 65.5 111.5 d - 58.5 3-5 96.0 158.0 | e a 98.5 6.5 125.0 230.0 f | 1. 84.0 3:0 95.5 182.5 ¢ a - 79.0 4.0 106.0 189.0 =) It will be observed from these figures, 1) the lime factor! for rice agrees nearly with that of other Gramineae which is between 1 and 2; 2) the rice plant seems to possess a relatively considerable resistance power against an excess of magnesium carbonate? since this does not depress the yield so much as the same excess of lime; 3) Rice-culture demands special attention to the proper ratio of lime to magnesia, since the maximal yield depends to a great degree upon the ratio 1:1. These facts induced me to examine the ratio of lime and magnesia in various soils-of Japan. The analyses of soils were kindly furnished by Dr. Zsuneté and his colleagues of the Geological Survey of the Japanese Empire, who have published a compendious volume on the composition of soils in Japan. These analyses were made in the usual style and do therefore not exactly distinguish between the readily assimilable amounts and the total amounts of lime and magnesia soluble in hot concentrated hydrochloric acid. In examining the long list of CaO analyses I have observed numerous cases in, which the ratio 7— would MgO 1 Bull, College of Agric. ‘Tokyo. Vol, 1V. No. 5. 2 At least in presence of manure of alkaline reaction ; an acil reaction of the manure would probably charge this behavior, On the Influence of Different Ratios of Lime to Magnesia on the Growth of Rice, LO! not be favorable for a maximal yield of rice. Many of these unfavorable cases are mentioned in the following table: , a Z 9 In 100 parts of 3 In 100 parts of Z fine dry soil, Zz fine dry soil. Locality. 5 Locality. | | vy = af ° CaO MgO | So: SNe CaO MgO a | = i + =) Co Kurosakaraura, fe Hasamamura, =F Hinog6ri, 5 0.376 0.986 Oitagori, S88 | 1.445 0.312 Hoki. a Buzen. OH | - | Kitayamatomura, | 2 Fujinemura, = 2 Tkumagori, = 0.308 0.888 Fujigori, dBi 0.389 1.784 Yamato. 5 Suruga. 25 _ hs x . = -) Mikagemachi, = Kanaokamura, | “5 | Mukogori, ~ 0.058 0.488 Suntogun, og 0.450 | 1.735 Settsu, 3 Suruga. Oo : UV - a meet Nishinemura, 2 Tatsukawamura, | = Tgugori, 5 0.412 1.913 Iwatagun, Sz 1.440 | 5.890 ar rg Ke Se aS 3 Iwaki. a Totémi. At | _ = ee 2 ee O | . =? 2 Nakanomura. 2 Onishimachi, Sas lishigGri, 5 Tanogori, Ge 0.778 2.189 Idzumo. & Kotsuke. AR - Lo Sea 2 (ag = = =a Nagayasumura, 5 Kitamura, a5 Nakagori, < Suchig6ri, ss 0.652 ~ )\)\..3.236 Iwami. 5 ; Totdmi. 2s a Oe . UJ ~ | o's o - | Nishinamura, gz Komamura, 3 3 Kamog6ri, = 1.618 6.307 Irumagori, 38 1.610 | 0.480 Idsu. a Musashi, zs O A (3) ' - wana 2 c< 3.0 Doshimamura, | = a _Meijimura, = | 2°5 Kawarumagori, 25 9.130 0.004. Minamiumibegori,| 4 § 0.575 O.112 Iwasyiro, a vy Bungo. =+ UO & af aM ™ Daikonjima, a Kamisannomiya- | 5 Yatsukagori, & 1,259 0 368 mura, Yamagori, | = © 0.391 | 0.019 G ae ; y 3 = ' - Idsumo, fea} Iwashiro. a : 5 | o ) Tsudamura, = Ogimura, is a5 Yatsukagori, & 0.295 0.923 Skusvgori, Soz 1.100 0.407 Idsumo, roa] Noto, Kae te Mes T . — SS mormura, ~ | ) Otamura, a} Nakag®ori, 2g 0.180 | o.S12 | Chichibugori, 3's 0.330 0.940 = . ~ = Iwami. roa) Musashi. Sa | cy | | 102 K. Aso. = In 100 parts of “4 a fine dry soil. Zz Locality. 5 Locality, 5 3 | cad MgO 6 = : A Takidamura, oe unease a . 4 Ns | 97 * > Aa gED z5 0.567 0.033 shikagori, a ie a Shimésa. < ee a) Selo = Nissakamura, 6 Nikaiddmura, is Ogasagori, $2 0.531 272 Yamabegori, = ‘TOtdmi. Og Yamato. = AS < Tokachihara, 3 Hiraidsumimura, = z 2.129 0.490 Nishiiwaigori, 2 Hokkai = ikuchu = okkaido, ra Rikucha. a Yoshiminehara, @ Osumura, 3 Iwasegori, 5 1.516 0.096 Shidagun, 2 Twashiro. 5 Suruga. Z Kikydgahara, = Yoshiwaramachi, = Higashichiku- z 0.448 1.423 Fujigun, 2 magun, Shinano. 5 Suruga, a Kugamura, 2 Sekimura, fa Katorigun, 5 0.706 0.004 Nagoégori, z Shimosa. 5 KKadsusa, e Omagarimura, a | Miomura, a Senhokugun, z 0.351 | 1.616 Abegori, 2 Ugo, z. | Suruga, a Nakazatomura, = Takatagun, z 2.075 0.461 Idsu., = In 100 parts of fine dry soil, CaO MgO 1.545 0.021 0.739 0.200 0.904 0.130 1.670 0.146 1.602 0.867 0.029 0.272 1.428 ‘OOLE UOdN visouUdRW pUL SUT] JO SOLA JUOAD)JIpP Jo ae | ¢ i t FA XALF Td : 66 aodvd OT “duonyur ayy smoys ozeid siy pf 4 3 £ OW I l l ()v ) Die LOA LT ODS aor Lie « On the Determination of the Available Amounts of Lime and Magnesia in the Soil. BY T. Katayama. It has been shown by experiments of O. Loew and W. May in Washington! and of K. Aso and T. Furuta? in Tokyo that the best development of a plant depends, other things being equal, upon a certain ratio of the amounts of lime to the amount of magnesia available to the roots. In order to reach a maximum harvest, it is necessary to determine the available amounts of lime and magnesia in the soil, and then to provide for the proper ratio between these two on the basis of the result obtained, by the addition of the calculated amounts of lime and magnesia compounds. The extraction of the soil with hot concentrated hydrochloric acid yields very probably more lime and magnesia than can be dissolved by the roots, while the proposed extraction with ammonium chlorid will yield in most cases certainly too small numbers.* Also, the size of the particles to be separated from the soil, previously to the treatment with hydrochloric acid, forms a very important question. Some authors separate all particles smaller than 0.5mm. diameter and treat that fraction with hydrochloric acid of 10% or also of 309% at the ordinary temperature, while others apply boiling heat. Furuta separated all particles smaller than 0.25mm. and treated this fine sand+silt+clay’ 1 Bul. No J, Bureau of Plant Industry Washington i901. 2 Bul, of the College of Agriculture, University of Tokyo, vol IV, No. 5. ’ Jmmendorf proposed recently boiling with } normal sulphuric acid for 30 minutes and to determine in this solution the easily available lime and magnesia, 104 T. Katayama. with hot conc. hydrochloric acid. I have tried the following modification : I. The fine earth, <0.25m.m was treated with hydrochloric acid of 5% at the ordinary temperature, in the proportion of 25g : 75c.c, for 24 hours. II. The fine earth, 1 o6o8¢ = | | OS NSas ae fs 0.603 ,; 7 0.474 5 } “ The pots for the cultures with the Kawasaki soil held 5 kilo, which amount of soil contained therefore 21.7¢ Ca O and 27.5¢ Mg O. In order to procure the desired ratios the following additions were necessary : * After this treatment with HCI. 200c,¢ of water was added and heated again to boiling for ro minutes, then left standing for 15 hours before the filtration, > Séderbaum observed recently that a hydrochloric acid of 29% extracted in 48 hours at the ordinary temperature only a part of the availalde plant nutrients, Lime and Magnesia in the Soil. 105 CVO 86.75 ee Sat 1 RROu- i No addition. II. Moore asia0 Ca O = 50.4¢ Ca CO, IY. io 2 60.8g Ca O = 108.6g Ca CO, EX: ‘eos Sage-Ca O = 157-7¢ Ca CO, The pots for the cultures with the soil from Komaba! held 4 Kilo, which amount of soil contained therefore 18.32 CaO and 14.9g Mg O. Hence the following additions were here required : i. se No addition. Ta: to- 11.s¢ Ca O = 21.8¢ CaCO, III. weo7 26.22 Ca O = 46.9¢ Ca CO EVs, ‘07 Biss CaO = 73.69 Ca CO, Now if my determination of lime and magnesia in the soils corresponds really to the available amounts, the results obtained with those soils limed in certain degrees, must agree with those obtained with cultures in quartz- sand or water, in which the lime and magnesia were present in the same ratios and in form of soluble salts. The soils were manured in the ratio of 10g ,of monopotassium phosphate, 6.3g potassium sulfate, 10g sodium nitrate for 5 Kilo. Sand culture. About 15 Kilo of pure quartzsand were left with conc. hydrochloric acid for 3 days, and then well washed with water, until every trace of acid reaction had disappeared. Five flower pots received cach 700 grms of this sand, and 14g of ® This soil from a depth of one foot was analysed some years before by 7. Furuta, while for my analysis and cultures served the surface soil only, hence some small discrepancies are easily accounted for. 106 T. Katayama. precipitated air dry aluminium silicate (29% of the sand) to increase the water holding capacity. The solutions applied had the following composition : Oni, KEI 0.30% K-N@: General Manure : 0.2% K,H,PO, trace Beso, Choe 0.3% CaO = 0.879% Ca (NO,), . 0.3, Mg O= 0895, Mg SO, (2) ae 0.4% Ca O = 1.417%, Ga (NODE ‘ 0.2, MgO = 0.596, Mg SO, (3) ne = 3 0.45% CaO = 1.318% Ca (NO,), 0.15 ,, Mg O = 0.447,, Mg SO, (4) es 0.48% Ca O = 1.406,, Ca (NO,), x o.12,, Mg O =ronsea; Meso, CaO 6) appa 0.5% CaO = 1.465 ,, Ca (NOx). 0.1 ,, Mg O = e2ohs ait ee The pots in each of the four series received 330c.c of the above mineral solution respectively. Since the evaporation of water from sand is rapid, a special arrange- ment was provided to secure a constant supply of water consisting in a band of loose cotton freed from the adhering fat, and immersed with one end in water. These bands encircled the pots; thus moisture due to capillary attraction was present always in the pots. For my experiments the growth of onion plant was observed in these soil and sand mixtures.1! 1 Experiments with oats, and bean and bukwheat ‘also had bean commenced but unfortunately insect pests and parasitic fungi caused so much damage that I was compelled to abandon them. a? Se ee Lime and Magnesia in the Soil. 107 Experiment with the onionplant in sand culture. 10 seeds were sown, on March 13, and the number of shoots reduced to five of nearly equal size on April 1. The height of the young plants was measured, April 29 with the following results : Ratio of CaO ; Mg O. I I 2 I 3 I 4 I 5 I 8.0cm 10. 5 Tek 8.2 8.0 7 S35 10, 2 7.0 8.2 73 7-4 » 9 8 6.8 Flot} 7-O OO) cp 8.6 6.1 Ta, 6.9 SOs 7.9 6.0 6.9 6.8 average 6. 84.,, 9. 30 6. 72 | 752 | 7.20 A photograph was taken, on May 1, see Plate XI This result shows the ratio pe ON as the best for onion. MgO Experiments with Onionplants in Soil from Kawasaki. Fifteen sceds of onion sown on May 2. The rate of germination was as follows : Date es =-2.) (o--+) oat) MeO 3 Gio I MgO ~ 1 MgO ~ 1 May 3 — 5 5 su LO 6 | 6 10 I4 ke S | 10 13 I4 108 T. Katayama. The number of the young shoots was reduced to 5 on May 15. The number of branches and the height of the young plants were measured on June 30, with the following results : Se Gan) Ratio MgO Number of branches Length, cm. a 4 22.5 b 5 29.0 0.75 ; : af I d 4 25-5 e 4 27.0 sum 21 average 25-3 a 5 35-0 b 6 36.0 2 c 5 34-5 : d 4 23.0 . 5 34-0 sum 25 average S355 a 4 31-5 b 4 31.0 : c 5 25.0 J : d 5 24.5 (s 5 29.0 sum 23 average 28. 2 1 4 27-4 b 5 28.0 AF c 4 26.7 I dd 5 32.0 € 3 18.0 sum 21 average 26. 4 $$$ — These onion plants were harvested on July 12, and yielded the following results: Rati Ca O Ratio Mg 0 Length of shoots, em Lime and Magnesia in the Soil. a 29. 40 b 35-70 — c 27.30 : d 30. 30 = 33-75 average 31.29 sum a 42. 90 b 44. 10 = c 42. 60 : d 39. 00 e 41.10 average 41.54 sum a 39. 00 b 36. 60 2 c 30. 60 3 x d 29. 70 e 36. 60 average 36.50 sum a 36. 00 b 36. 60 4 G 33- 20 I d 37. 80 e 21.90 average 32.50 sum Number of shoots Gi. OBR ke wGr on Total weight, g. oO t. °o to ay) [e] sum 14. 40 4. 60 4.45 sum 25-51 4.05 4. 00 2. 60 3-15 4.50 sum Id. 30 st = wv I1IO T. Katayama. Experiments with Onionplant in Sotl from Komaba. Fifteen seeds of onion were sown on May 2. The rate of germination was as follows: The number of young shoots were reduced to 5 on May 15. The number of branches and the height of the young plants were measured on June 30, with the following results ; Number of branches Length, cm average 25.2 ih average 33-5 Lime and Magnesia in the Soil. Number of branches, Length, cm. average 33-0 ~ 1CaO Ratio Mg O to average 39.78 sum 8. sum 24 sum 24. Number of shoots. Total weight, g. ih, T. Katayama, Ratio aa Length of shoots, cm, Number of shoots. Total weight, ¢. H 41. 40 5 5:20 b 39- 90 5 4. 80 3 c 41. 40 4 3-75 ; d 37. 80 5 3-45 € 36. 00 5 275 average 39.34 sum 24 sum —- 19. 95 a 36.6 5 3-39 b 36. 6 5 4.10 c 42.0 5 3°95 OF d 37.8 4 3.10 e 37.8 5 3. 80 average 38.01 c sum 24 sum 18,25 Hence we find in all these cases that the best ratio of aa or the lime factor for the onionplant is = 2. The chief results with two different soils agree therefore very well with the results in sandculture, where the total content of lime and magnesia was certainly present in an easily available form. It may be safely concluded that the modification I proposed to determine the available amounts of lime and magnesia in soils is in close agreement with the actual results and furnishes therefore a reliable basis for the calculations regarding the liming of soils. It might be objected that the conditions of the absorptive powers in certain soils might alter the availability of lime and magnesia for the roots. But in my investigation the two soils applied differed very much in character and nevertheless yielded results which agree with each other and with those of the sand culture. Only soils with a unusually high percentage of clay or humus might yield results differing somewhat from my expectations, but such soils are from the outset not favorable for agriculture. The experiments of the year 1902 just mentioned were repeated in Lime and Magnesia in the Soil. 113 1903. The same pots served for this second series, the soil was not renewed, but it was manured again uniformly with 20g KH,PO, and Is5¢ (NH,),S5O, for each 5 kilo. The pots were kept in the green house. Second experiment with onton plants ix the soil from Kawasaki. I5 seeds were sown per pot, on January 7, and the number of the young shoots reduced, on March 2, to six per pot, all being of equal size. The height of the young plants and the number of branches was measured, on May 1, with the following result: a Ca Ratio MzO Number of branches, Length cm. g a 4 22.0 b 4 33-9 c 4 29-5 0.75 “eo d 3 30.5 I e 5 27-0 f 4 34.0 sum 24 average 31.0 a 4 33-9 b 4 32.5 Cc 5 46.0 2 a: d 5 32.5 e 4 31.5 f 4 34.0 sum 26 average 34.9 a 4 34.0 b 4 32.5 c + 30.5 3 —— d 4 35.0 : 3 e 5 33.0 3 4 29.0 sum 25 average 32.7 114 T. Katayama. a E Ratio Meo Number of branches. Length cm, | a 4 30.0 b 4 30.5 c 3 25-5 oor d 4 32.5 I e 4 29.0 - 4 35-5 sum 23 average 30.5 Caen n ee eee eee een Up to this time, almost every day 200CC water for irrigation was applied, but the quantity was gradually increased to 300CC. On harvesting, on June 7, the following result was obtained: reer ———————o———EEEEEEEEEEEEEEE———— Ratio areas Number of branches. Length cm, Total fresh weight, grm, a 6 58.0 2215 b 6 61.0 Dyes Cc 5 57-0 17.0 0-75 d 6 57-0 30. 5 ; e 7 52.0 14.5 f 6 62.0 22.0 sum 36 average 57.8 sum 124.0 a 6 59.0 24.5 b 8 58.0 31.0 c 7 74-0 49-5 2 d 8 57-0 19. 5 ; c 7 60. 0 28.0 f 7 61.0 25.5 43 average 61.5 sum 178. — a 7 60.0 29.0 b 6 62.0 24.5 c if 63.0 25.0 i d 7 59.0 31.0 ec 6 63.0 25.0 f 6 45.0 by Pl 39 average “58. 6 sum 152.0 Lime and Magnesia in the Soil. II5 Ratio su Number of branches. Length, cm. Total fresh weight, grm. a 5 55-0 25-5 b 6 57-0 25-0 c 7 60.0 22.0 = d 6 55.0 22.0 ; e 5 52.0 16.0 - 7 60. 0 28.0 sum 36 average 56.5 sum 138. 5 These figures, as well as the photograph taken shortly before harvest- ing and reproduced on Plate XI show plainly again the superiority of the ae oa G) 2 t : ratio Mea for the onion plants. Second experiment with onion plants in soil SJrom Komaba. 15 seeds were sown into each pot, on Febr. 10, and the number of the young shoots reduced, on March 20, to six of equal size. The height of the young plants and the number of branches were measured, on May 12, with the following result : oo eeeeeeeeeeeSSeSeSeSeSeSeSeeeeeSSSFSFSseseseF mecca O . Ratio Number of branches. Length, cm, Mg O < ee a 5 36.0 b 5 i7.o c 3 37-9 : 1 4 26.0 € 30, € i é e 4 40.0 r 4 37.0 sum 25 average 37.8 ee ee Bk | 1 6 47.5 b 5 40 oO ¢ 5 45.0 <<. a 5 45.5 1 e€ 6 43.0 t 3 38.0 3° 43.1 116 T. Katayama. Tl Cai@ Ratio “MgO Number of branches. Length, cm. a 5 45 b 3 40 c 5 38 == d 5 40 e 4 42 f 3 38 sum 25 average 40. 6 a 5 40.0 b 5 41.0 c 5 39-5 oa d 4 37-5 e 2 42.0 1k 3 40. O 25 A photograph was taken shortly before harvesting, on June 15; it is reproduced on Plate XI. On harvesting, the following result was observed : Ratio Se Number of branches. Length, cm. Total weight, grm. a 5 45 18. 5 b 5 48 18.5 c 4 47 12.5 —. d 5 45 10. 5 e 5 49 15.0 f 5 47 8.5 sum 29 average art vy sum goes a 7 63 28.5 b 6 53 19.0 c 6 58 22.5 a d 6 58 25.8 e 6 55 22.0 f 5 5 17.2 sum 36 average 57.2 sum 134.7 Lime and Magnesia in the Soil. 117 Ratio aes Number of branches. Length cm. Total weight grm. 59 19.0 53 15.5 50 19.0 3 50 18.0 : 50 21.0 53 18.0 average 52.5 sum 110.5 5° 19.7 52 21.0 54 23.0 : 53 17.0 55 16.0 45 16.0 average 51.5 sum 112.7 Also this result shows that the lime factor for the onion plant is = 2. Second series of Sandculture. A large amounts of sea sand (particles smaller than 1mm. diameter) was soaked in conc. HCI for about a month, and well washed with distilled water until every trace of chlorine reaction had disappeared, then left to dry. Five pots received each 1.5 litres of this air dry sand, and 8oc.c of nutritive solutions which had the following composition : 0.2% KH,PO, Geb anys Na, SO, a a 0.01 ,, Fe SO; he 0.3 9/ Ca O =.0.8799% Ca (NO,), 0.3 » Mg O = 1.098,, Mg (NO,), +a (2) rat =— 4% CaO = 1.117% Ca (NO,). 118 T. Katayama. — 0.459 CaO = 1.318% Ca (NO), : 0.15 ,, MgO = 0.549,, Mg (NO,), +a So 0.48% 'CaO = 1.406% CaINOam i 0.12 ,,M¢g O = 0439,, Me (NO,)2 4a (ages 0.5 % CaO = 1.465% Ca (NO,), Me O I : 0.1 ,, Mg O = 0.366,, Mg (NO,), +a At the beginning of this experiment, the total concentration of the mineral nutrients for each pot was 2.14—2.38 per mille. The pots were kept in the green house. 20 seeds of onion were sown, per pot, on March 6, and the number o young shoots reduced, on April 7, to 8 of equal size. The height of the young plants and the number of branches was measured May 1 with the following result: Ratio aie Number of branches. Length, cm, a 4 21.5 b 5 23.0 c 5 25-5 d 5 23-5 ; e 4 25-3 | f 3 18.5 g 4 22. 7 h 4 DT 2 sum 34 average 22.6 \ 5 30.1 b 4 24.0 c 5 27 $ d 5 26. 2 2 : € 4 24.9 f 5 25.0 g 4 28.9 h 5 26, 2 sum 37 average 26.6 aera > Lime and Magnesia in the Soil. 119g LL Ratio pes OF Number of branches. Length, cm. Mg O a 4 25.2 b 4 27-5 c 4 20. 5 d 5 Zens 5 e 5 18.0 I f 5 26.5 9 g 4 24.0 h 5 23.9 sum 36 average 23.4 Pe eee eee a Is 26.0 b 4 20. 2 c 4 21.5 d 5 25-5 4 e 4 28.3 I f 5 28. 5 $ 4 22.3 h 5 18. 6 sum 36 average 23.9 —————————————————————e a 4 18.2 b 5 21.7 “ 5 26.5 d 4 26.1 2 e€ 4 22.0 I x 4 17.0 g + 18.3 h 3 18. 7 : sum 33 average 21.8 The mineral solutions above mentioned were added in the following proportions : a 120 T. Katayama. Date. March 9 | April 20 May 15 June 7 Quantity. Fs (CAC | 30 40 C.c 40 C.c In the begining of this culture, every day 25c.c distilled water for irrigation was applied, and the quantity was gradually increased to 7oc.c, but sometimes to 100c.c in very warm weather. The plants were harvested June 27, and the following results observed: Pp U. 5 Ratio aoa Number of branches. Length, cm. Total weight, grm. a 8 30. I . 10. 8 b 6 aie S22 c | 25.5 10.7 d 4 34.0 6.3 : e 6 37-2 10.3 f 3 22.3 1.9 g 5 26.1 ae h 5 6.9 Lick sum | average 29.2 sum "Goan a 7 30. 2 9.2 b iS 30. 0 .8 c 7 40.3 9.5 d 7 83.0 10. 0 . e 7 35.2 Eioe f 5 35-3 9.0 g 6 37-4 6.5 h 7 38.0 11.6 sum eet average 34. 9 7 sum 73- 9 U 7 22.1 6.3 b 6 30. 2 10,5 c 5 24.7 ony d 7 35.0 10. 7 i ¢ 6 30. 5 8.8 f 5 38. 1 8.9 g 4 BPG 6.5 h 6 28.9 6.4 sum nai, average 29. 7 sum 66.8: ea. - Pot i Lime and Magnesia in the Soil. I21 Ratio eur Number of branches. Length, cm. Total weight, grm. a 4 30. 3 4.7 b 5 23-5 5-7 c 7 273 7.8 d 7 30. 4 9.0 4 e 8 31.0 8.4 f ) 29. 2 roe g 29.0 10, 2 h 7 28.0 My bee sum 51 average 28.6 sum 64. 5 a 7 29. I 9.7 b 8 29.6 [252 c 6 aig 8.7 d 4 27.0 5-3 5 e 6 23..2 I~ 2 if 4. 29.5 5-7 g 6 26.1 5 Be h 5 30. 2 8.0 sum 46 average 28, 2 sum 63.9 Additional Experiments. Experiment with pea in the soil from Kawasaki. All principal conditions were the same as in the first onion experiments above described. 15 seeds of pea were sown, on Febr 9 and the number of young plants reduced to five of equal size on March 18. The formation of flowers commenced on May 3 and ended on the 24th of the same month. Up to the flowering period, almost every day 4ooc.c. of water for irrigation were applied but later on the quantity was increased to 600c.c. | When the fruits had reached the ripening stage, on June 7, watering was stopped and {22 the plants left to dry. result: T. Katayama. The harvest on June 15 yielded the following number of Rati ECa@ average weight of weight of ee Mae length. branches. | fresh fruits. | air dry seeds. Bed 98 cm 7 18.3 grm 15.7 grm I 2 : 115» ) 33-0» 27-5 35 2 120s sf) 38.3 BUR I 4 1250s. 9 38.5 31.0 weight of air dry air dry straw,] total weight. 11.8 grm 29.0 grm 1 Onmes 49-3 55 21.5 58.0 20-5 57:3 Experiment with pea tn soil from Komaba. The number of plants, time of sawing and harvesting was the same as in the case just described. On harvesting, on June 15, the following results was obtained : Pad Ca@ number of average weight of | weight of | weight of air dry length, branches. | fresh fruits. | air dry seeds.lair dry straw.| total weight. | 120 bs : y 35-5 grm 29.7 grm 18.5 grm 52.8 grm D2 ee II 38 s 31.8 23.0 59.4 Sy] as II o ae ; 33.9 a ‘ 66.7 pet ee ; II en Bie 22.6 Th 58.7 3oth results of experiments with different soils show that the ratio sae is the best for the pea. Experiment with oats in soil from Kawasakt. 15 sceds were sown December 15, and the number of young shoots reduced to six all of equal size, on March 6; all conditions were essentially the same as in the former cases. Lime and Magnesia in the Soil. On harvesting (June 29) the following result was obtained; O ‘ NT. es 7° as f Ratio Seu Average height. oe W ae of Total weight. eet : ; = 94. 26 41.4 118 105 28 46.1 143 ~ 103 28 45.7 141 ; IOI 26 42.9 | 115 Experiment with oats in the soil from Komaba. These experiments were made at the same time as those with onion. The result was as follows ; Ratio ee Average height. Number of Weight of } Total weight. stems. seeds. DUZ 21 40.1 99.5 117 24 43.3 113 114 25 43.8 117 107 18 VAS 103 Both results of experiments with oats in different soils show that the ratios and oad é : 1 In order to procure the ratio —, this pot received 15,9 gr MgO = 33.1 gr 3 2 In order to procure the ratio 2 ~ I are the best. > this pot received 21,7 gr Mg O = 45-2 gr 124 T. Katayama: Lime and Magnesia in the Soil. Conclusion. It will be noticed from the second series of experiments with onions, that the results fully confirm the results of the first series that is: Sand- culture as well as the cultures in two soils differing widely in character from each other yielded the best results when the available amounts of lime and magnesia were present in the ratio 2: 1, in other words the onion has the limefactor 2. Lime and magnesia in the sand culture were added in form of solutions, hence the total amount of these salts were easily available even if precipitated as finely divided phosphates. As to the soil culture the “available amounts” of lime and magnesia were determined according to my modification of the usual method and their ratios changed by adding carbonate of lime in such quantities as to reach the fixed ratios of the sandculture. Since in all my experiments of 1902 and of 1903 the ratio Siro proved the most favorable for the onion plant, the determina- tion of the available amounts must have been made by a reliable method. Hence my modification of the usual determination of the available amounts of lime and magnesia may be stated again: I propose to separate all particles <0.25 m.m, to determine the percentage of this fraction, and to extract this fraction for 50 minutes with boiling hydrochloric acid of OF Fi aye 4 10% in the ratio of 25g : 5o0c.c. oe ke BUL. AGRIC. COLL. VOL. VI. rime ec! 100. To page a == ~ is = = — = \A — — ~ C i o » To page Komaba. experiments of 1got. Soil of ® ‘ i ' $ + F = = Fy . ' . i ‘ Rae ~ Pt . ~ ‘ , p ‘ “ ' ‘ : * aw ~s a Es . a * 4 — : ’ * Pes ‘ # i ae i - ” > . : ‘ y ‘ ? wy » ; io |8 - rs 1 e ~~ b — ‘ , ‘ : - 1 -_ 7 5 ‘ > ; , + 7 7 - : » i ond MW . - ahr. 2 - nd awe = ——— = 7“ as — = a ime 3 = ae Ueber den Einfluss des Mangans auf Waldbaume. VON Oscar Loew und Seiroku Honda. Da Manganverbindungen einen giinstigen Einfluss auf landwirtschaft- liche Gewiichse Aussern,! war es wiinschenswerth, auch tber den Grad des Einflusses auf Waldbaume einige Anhaltspunkte zu gewinnen. Manganoxyd ist schon haufig in der Asche verschiedener Holzer, in Blattern und Friichten verschiedener Baume gefunden worden, die Mengen desselben variirten aber je nach dem Standorte ebenso wie die Mengen des Eisenoxyds betrachtlich,? wie die folgenden Daten, welche mit Ausnahme der letzten Zahlen den Aschentabellen Wolff’s entnommen sind, erkennen lassen: Procente in der Asche. Object. Seo ass aS See Analytiker, Mn, O, Fe, O, Weidenholz 0.15 0.53 Reichard Birkenholz I 3-94 3.00 Wittstein Birkenholz II 4.13 0.59 Berthier Lindenholz 0.88 0.15 9 1 Siehe diese Bulletins, Bd. 5. 2 Nach Wodff (Aschenanalysen, 2 Teil, S. 159) nimmt der Mangangehalt der Baume zu, wenn es an Kalk mangelt, was von Councler bestiitigt wurde (Zeitschr, f. Forst=und Jagdwesen, Bd. 14, S. 113 [1882] und Bd. 35, S. 391 [1903]. Allerdings beobachtete letzterer auch, dass in den kalkreichsten Organen (Nadeln) der Waldbiiume auch der Mangangehalt grésser war als in den anderen Theilen der Baume. 126 Oscar Loew und Seiroku Honda. SS eS ee eee Procente in der Asche, Object. ——_—$—_—_—_—_—- Analytiker, Mn, O, | Be, 0; jt i Fichtenrinde 0.65 é 2.67 Wittstein Nirschbaumrinde 1.02 009 Palm Ulmenrinde 0.68 0.90 Zeyer Buchenblitter I Tires 1.07 Fresenius Buchenblatter IT 1.86 | 12.00 Wittstein Weidenblitter 0.29 : 0.96 Reichard Birkenblitter 673 1.14 Wittstein Kastanienfrucht 5.48 5 HOP Richardson Buchensamen 3.10 2.66 Souchay Fichtenpollen Rote 1.95 Ramann Man ersieht hieraus, dass der Mangangehalt nicht selten den Eisengehalt weit iibertrifft, Dass dieser Mangangehalt irgend einen Einfluss auf die Waldbiume dussern k6énne, war unbekannt; man_hielt ihn fiir unwesentlich und fiir zufallig aus dem Boden aufgenommen. Wir wiahiten zu unsren’ Versuchen die fiir Japan so wichtige Cryptomeria japonica. Am 29 April 1902 pflanzten wir auf Beeten von zwei Quadratmeter Grésse die jungen nahezu gleichgrossen Baume ein. Zu dem Versuch dienten sechs solche Beete, welche von einander durch eine etwa 1 Meter breite mit niederen Brettern abgegriinzte Fliche getrennt waren. Der Boden’ war ein humoser Lehmhoden von nur massiger natiirlicher Fruchtbarkcit. Jedes Beet erhielt anfangs neun Pflanzen, die aber spiiter auf acht reducirt wurden, da einige eingiengen- Auf Beet No. 6 waren nach ciniger Zeit nur siecben Pflanzen erhalten. Die Beete wurden genau wie alle jungen Pflanzungen der Férstereien behandelt und im Winter gegen Frost geschiitzt. Die Loesungen wurden nicht gleich- massig tiber die Beete verbreitet, sondern in eine kleine Vertiefung um jeden Stamm die berechnete Menge gegossen. Die Loesungen wurden zehnprocentig vorratig gehalten und die jedesmal notige Menge auf das hundertfache verdiinnt. Ueber den Einfluss des Mangans auf Waldbaume. 127 Beet No. 1 erhielt von 1 Mai bis 1 November 1902 allmonatlich o.5¢ Mangansulfat, nur bei der ersten Begiessung die doppelte Menge; im Jahre 1903 aber vom 1 Mai bis 1 Nov. (inclusive) allmonatlich 1g. Es erhielt also jede der 8 Manganpflanzen im Ganzen 1.5 gramm _ jenes Salzes. Beet No. 2 erhielt Eisensulfat (Eisenvitriol) in gleicher Weise und Menge wie Beet No. 1 das Mangansulfat. Beide Sulfate waren die krystallisirten, nicht chemisch reinen, Producte des Handels. Beet No. 3 erhielt lediglich die jenen Loesungen entsprechende Menge Wasser, und diente wie Beet No. 5 und No. 6 als Controlbeet. Beet No. 4 erhielt Kochsalz, No. 5 Natriumnitrat, No. 6 Calcium- nitrat in denselben Mengen wie Beet No. 1 das Mangansulfat. Diese Nitrate wurden angewandt, um zu beobachten, ob eine teilweise Diingung eine 4hnliche Wirkung aussern kénnte, wie Mangansulfat. Das Chlornat- rium auf Beet No. 4 sollte Aufschluss geben, ob die Holzbildung beférdert wird ; denn in der Landwirtschaft ist bei einem gewissen Chlornatrium- gehalt des Bodens ein schnellerer Verbrauch von Starkemehl und Zucker zu Gunsten der Ausbildung der Holzfasern beobachtet worden, oder es wird wenigstens ein derartiger Einfluss des Kochsalzes fiir wahrscheinlich eehalten. Vier Monate nach der Behandlung war noch kein deutlicher Unterschied im H6éhenwachstum wahrzunehmen, erst im fiinften Monat war ein Voraneilen der Manganpflanzen deutlich zu erkennen. Dieses Voraneilen nahm aber im folgenden Frihjahr ein rascheres Tempo an, bis im Herbst des zweiten Jahres ein hodchst auffallender Héhenunterschied erreicht wurde, wie aus folgender Tabelle ersichtlich wird. Die Zahlen geben die Héhen in Centimetern an. Osear Loew und Seiroku Honda. 128 — | — if! Sel) Gn gS OC: |e Grex £6 Oe |) vor ger |e o2en lS ex 6g S‘bz | Loz 8 1, o6z | ror | 26 Sof | ogr | of Obes | Gor |) a6 GSe, | LS gor (OEZe | eigrey Zer |SSSe | eer L for || L:z€ || o'oz Te | @2Gensl ain [6 Oe. |) Sigr 66 oof | £07 g6 GTS, ||| eee hix | g2e | S312 9 (Gt |] toytets || oyter 96 Osea) soz 18 Ste || 4°91 Sor | gf | Sor oye i (ence Sy ea Gor | fore |) Si6x S mat |] erAS ol orttrs cyl || sSeT pal) olor 09 of | gli OHI 5 |n0y4z0)|) lor [6 Baise |) Waite Spr | ob | ot + S9 OM Selon z9 Gee | Ogi c6 o'6z | 9°61 oY o61 | Zor Sone || RAS | Woe €f1 | oF€ | L61 ¢ bS bre | S:oz gor | oct | 661 L6 oof | Loz 6g Oem pac or Sg | of O'7z gor | Lb | S61 Zz | OLIe || PROeml eh con | al 4 Oe eet wie Os OMe || SOT Cer eee see Peek Sit ||-S6z), | 4:6ra) Str | S6r | og I | | oe ee fOr OT Cot oor So lon 20/0, CO 80/55 CO) or 0], MU OT oT hd C0/o, zo c £0 lor | é or | zo bs “AON, | “AON, IVIL “AON, | “AON Iv] “AON, | “AON | TRI “AON, | “AON, | IVT “AON, | “AON Iv] “AON, “AON, Iv] azuepg op ara! | . pags 2 ‘be ¥Q¢ 92; ‘be $+ os uy ; }VAPLULUN TO] Vo) PETVIUUAOLTY © NT WOT VUIOTYS) JO-4uUuo_) ae eancnteee Ueber den Einfluss des Mangans auf Waldbaume. 129 Hieraus beerchnet sich fiir die Zeit vom 1 Mai t1go2 bis to Nov. 1903 das durchschnittliche Zuwachsprocent bei Behandlung mit : Mangansulfatiziv-.6 one) 1.226 lve 858.7 Bermmaswiiat nih {qsrio, tie. bol. 445.5 Phiematmemiy linseed. Lb. 2. . 34555 Natriumnitrat ... . ee: a A26:7 Malcincmitrat wet ks... wast. \..2° 340.6 und fiir die Controlpflanzen ... 448. ve) i) Man erkennt hieraus, dass Mangausulfat das Hoheniwachstum stark Jorderte, dass das Ferrosulfat diese fordernde Wirkung nicht hatte und Chlornatrium sowohl als Calciumnitrat hemmend gewirkt haben.! Be- merkenswert ist noch, dass die Diingung mit Chilesalpeter das Héhen- wachstum nicht fo6rderte. Indessen der Ezsenvitriol sowohl als der Chilesalpeter haben das Wachstum der Zweige begiinstigt wie aus dem Vergleich der Totalgewichte erhellt. Am 17 November wurden die Baume am Grunde abgesagt und frisch gewogen mit folgendem Resultat bei: Manganosulfat ® ... ... 5874 gramm. Betesmiabe “2 Sac A. 3 = ; aes 8 Pflanzen. Ghioriatruim.. .... «.. $300 a Wathigmmitrat... 9... ... 3355 Calciumanitrat 2... ..., 2497 » (7 Pflanzen) POM pies css Yes nee, 2535 » (8 Pflanzen) Es ergibt sich daher als Durchschnittsgewicht ftir eine Pflanze: Det Manicanosaiet.F 55°... ... 733.8 ¢ PECREGHEMIAE aaah ey cnc ks | 4245 en SeieotnettroMm se vs §=6173,8 “Sp eglag leurs coe a ry, © Ce PC ONGIOMOTRERQG deus) uc «ee cvs. 356-7 ba RC ORGMOR-\ Lent kee, hs ~ 316.9 2 Auch Loughridge (Calif. Stat. Bul. 133) und Aossowitsch (Journ, f, Exper, Landw 1903, p. 44) beobachteten einen sehr schiidlichen Einfluss von Kochsalz auf Biume, Natriumsulfat ist nach diesen Autoren bedeutend weniger schidlich, 130 Loew und Honda: Ueber den Einfluss des Mangans auf Waldbianme. Die Durchschnittshohe bei der Pflanzung der Baumchen war auf dem Manganbeet=19.4, auf dem Controlbeet aber=17.1 cm. Berechnet man nun das Erntegewicht bei den Manganpflanzen auf gleiche Anfangshéhe wie bei den Controlpflanzen um, so hat man fiir eine Manganpflanze=646.7¢. fiir die Controlpflanzen gleicher Anfangshohe= 316.9¢. Die Manganpflanzen hatten also fiir dicselbe Anfangshihe nach i} Fahren die Controlpflanzen um das doppelte (2.03 fach) an Massenzunahme iibertroffen, wie auch wohl aus der in Tafel XII reproducirten Photo- graphie abgeschitzt werden koénnte.! 1 Von einigem Interesse ist noch der Unterschied in der Wirkung von Natriumnitrat und Calciumnitrat, weil solche Unterschiede zu Gunsten des Natriumnitrats auch in der Landwirtschaft beobachtet sind. Es wirkt eben auch das Natron in der Form des Nitrats stimulirend, wahrend Chlornatrium in Folge des Chlorgehaltes wieder hemmend wirkt, wenigstens in grésseren Dosen. In relativ geringerer Menge kann aber auch dieses eine missig stimulirende Wirkung auf Feld- gewiichse ausiiben. Fichten reagiren nicht so energisch auf Mangan als Cryptomerien, Versuche in grésserem Massstabe wird der eine von uns (Honda) weiter fiihren, BULL, AGRIC, COLL, VOL. VI. PLATE X/JiT. Control plants. Manganese plants. Plate showing the stimulating effect of manganous sulphate upon the growth of Cryptomeria japonica. To page 130. On the Practical Application of Manganous Chlorid in Rice-culture. BY K. Aso. In the last volume of this Bulletin, several communications regarding the stimulating action of manganese upon plant-growth were published. Loew and Sawa! observed this action with plants in water and soil culture and also the author? with various plants in waterculture. Nagaoka further carried out a field experiment with rice in wooden frames and obtained an increase of one third of the harvest in grains by the application of man- ganous sulphate at the rate of 35 kilo Mn, O, per hectar. In all these ex- periments, mangano sulphate was used. But, since manganous chlorid is a cheap by-produrt in the bleaching powder factories, it seemed to me of some practical importance to make also an experiment with this salt. My experiment was carried out in the paddy field with rice in quite the same manner as the practical farmer does. Two square-shaped plots, each of 30 sq. metre, were selected ina field which had not been manured for several years. Each plot received 27 kilo barnyard manure, 15.5 kilo rotten human excrement, 230 grams double superphosphate and aftwards 570 grams wood ash. Besides, one plot received 200 grams crystallized manganous chlorid* (corresponding to 25 kilo Mn, O, per ha.), while the other served as control. On July 3, the young rice plants from the seed-bed were transplanted, 1 Bul. College. Agric. Tokyo. Vol. V. No. 2. 2 ibid. No. 2. 3 ibid. No. 4. + This was applied separately after manuring. 5 The variety was the Satsuma. 132 K. Aso: each plot receiving 305 bundles of twelve equally developed individuals. The irrigation-and the drainage were made in each plot separately in the same manner as practically carried on and care was taken to avoid the passing of drainage water from one plot to the other. Towards the end of July a difference in regard to the development was quite marked and became gradually more noticeable. On September 3, all plants in the manganese plot flowered and four days later in the control plot. The weather conditions were very favorable for rice culture through- out the whole summer, and all possible attention was paid to avoid damages by insect pests. On November 6, the plants were harvested. The harvest was weighed in the air dry state: Manganous chlorid. Control. RotalGheaey est @ tics: eta oes wegen ater 2257 ak 16:73) ke Sa a Sait ea AG W RCT EE ce, ee 12.00} 4s. 8.19 k. SR GitalerGaiins ohana cee eden cee nis eee 11.49 k 8.46 k Bull Cains 25.06 s¢icc<2-8 Oe OR ae 1 L230 8.23 k Bin pty eras) ileseuniins gf. ....anele. pee 6.26 k @:32) FIUSKEG MUNI SOGAINS coc cuenenancccteetee teetaeeate 8.66 k. 6.65 k. Weight of 1 Litre of unhusked full grains. 616 g. 619 g. Now, if the yield of the control plot is taken as unit, the following figures are obtained: Manganous chlorid plot. Total yieldgda.. Sin. isi’ he aie pe amie pea tae ae 1.42 ro A ge et ae ee ee ania Peamentene 1.48 Full grains: (uihusked)........wateasie ose ehes 1.36 Parl opine (RS OGY ise es act's ana dca Skewes 1.30 On the Practical Application of Manganous Chlorid in Rice-culture. 13 oP) These figures show an increase of one third of the grains by the appli- cation of 25 kilo. Mn, O, per hectar in the form of manganous chlorid,’ which is in full coincidence with the result of Prof. Nagaoka who had applied the same amount of Mn, O, in the form of the sulphate. Sinee the area of one plot corresponded to 34, hectar and the quantity of manganous chlorid applied was 200 grams, 66 kilo of this salt would be required per hectar. The cost would be only 4.4 yen,? while the value of the increased harvest is 137.33 yen.* There experiments will be continued for a series of years on the same plots. 2 As to pot cultures manganous sulphate would be preferable to the chlorid, since chlorids often exert an injurious influence on the yield. In field culture—especially with paddy field—this depressing factor is removed by rains and irrigation water. The manganous chlorid of course changes in the soil into other manganese compounds. 2 The price of roo pounds of the crystallized salt Mn Cl, +4 ag. is three yen (about six Mark). 3 Compare Bul. College. Agric. Toky6. Vol. V. No. 4. p. 472. On the Stimulating Action of Manganese upon Rice, II. BY M. Nagaoka. In a former Bulletin (vol. V, No. 4) I had shown that manganese com- pounds can increase the yield of paddy rice. The experiment was repeated under essentially the same conditions in the same frames as before, the only difference being that no fresh doses of manganese sulphate were applied since this time it was the chief object to ebserve any after effects of the first doses given the previous year. The crop was harvested on Nov. 11 and weighed two months later after being well air dry. The results are seen from the following table: Full Empty Average. No. of Straw grains grains | Total Frames. Full Empty Gee ke. er. gr. gr. grains. grains. ie sida I No manure iy 2a) | 217.9 / ne and no i 4.0 288.1 186.7 3.4 272.0 462.1 ] | 25 Mn,O, ; 4.0 309.9 2 |NoMn,O, é 3.1 271.9 14 3:3 295.3 S076 5)" 35. 265.7 1) S068 | 26 4.2 319.9 | re ert: f 3 3:7 293-1 15 3.7 277-4 203.7 | 3.9 289.8 497.4 / 27 4.4 298.9 4 3-6 305.4 16 4.0 289.9 224.9 4.0 304.6 | 533.5 28 136 M. Nagaoka: On the Stimulating Action of Manganese upon Rice, II. Mn,O Full Empt Average. No. of a as Straw || e per ha grains grains Total. Frames. Full Empty Shee kg or or sr ns 5; raw. g. gr. or. gr. grains, grains, 5 238.0 4.1 323.4 17 20 248.0 4.8 219.4 || 230.3 4.4 316.4 551-1 27 205.0 4.3 306.4 | 6 244.0 5.1 324.8 18 25 238.0 4.8 S156 235.5 4.9 308.8 549.2 30 224.5 47 285.9 || 7 247.4 2 329.9 19 30 251.7 4.9 324.2 242.7 5-0 322.2 569.7 31 220.0 4.9 312.4 8 196.5 4.6 295.9 20 35 270.0 4.1 339-7 231.2 4.6 313-5 549-3 The harvest in full grains was therefore greatest in the frame that had received manganous sulphate at the rate of 30 kilo Mn,O, per ha; very near to this comes the frame with the ratio of 25 kilo Mn,O, per ha, which in the first year yielded the greatest weight of full grains. The increase over the manured frame without manganese was 16.9% while the maximum in- crease in the first year was 37%. On the Influence of Manganese Salts on Flax, Y. Fukutome. In former experiments, described in the Bulletins of the College of Agri- culture, was shown that manganese salts can exert a stimulant action on various plants serving as food. It seemed interesting to make further obser- vations also on plants that are cultivated for their fibres. I selected for this purpose flax and compared here the action of man- ganous chlorid with that of ferrous sulphate and Cobalt nitrate. The soil came from the experiment grounds of our College of Agriculture. Each pot containing eight kilo soil was manured with 16 g. superphosphate, Io ¢. potassium sulphate, 8 g. each of ammonium sulphate, and sodium nitrate. 40 seeds were sown in each pot on September 21 and the shoots singled out in October to 15 of equal height (=5 cm.). Pot I. had received o0.4¢ crystallized manganous chloride (Mn Cl,, 4H,O) Pot. If. 5 0.4¢ ferrous sulphate (Fe SO,, 7H,O) Pot III. ‘f 0.4g cobalt nitrate (Co(NO,),, 6H,O) Fo. IV. iy O.G2E |) 44 3 3 Pou. V. a o.4g (MnCl,, 4H,O)+0.4g (Fe SO,, 7H,O) Pot VI served as Control. On Nov. 30 the plants were measured and a photograph taken, repro- duced on Plate XIII. It shows that a stimulating action had taken piace. On Dec. 21 the stem and branches were again measured, whereupon the plants were cut and left to become air dry. At that time only two flowers opened, one had in pot IV and one in pot V. The results were as follows: 138 ¥Y. Fukutome: On the Influence of Manganese Salts on Flax. Nov. 30 Deer 2x eee Height, Number Number Average c cm. cm. of of length of ipheoee 8 average average Buds branches | branches Karery) IBS ea ees, ae 46.9 65.7 2 19 26.7 1LO:7 1 es Sk Mee oie es 47.4 67.8 5 LZ 24.5 11.6 WiCo (eve. 48.6 68.5 3 19 27.4 iG ®) PVC 0) (:02'2:):: 48.9 69.6 5 19 33.8 12.8- V.Mn+Fe ... Fhe 7, Wise 4 19 32.9 120) Vis Controls... 43:4 60.1 fe) 19 Pa 10.5 This result shows that the joint application of iron and manganese had a marked effect on the growth, while each alone but little in the dose of 0.4 g¢ per 8 kilo soil. Also cobalt nitrate in the small dose of 0.02 ¢ per pot exerted a stimulating effect. —____—_——al>—9-c-.2sss2-2eeca. eee 3 g. One pot received 10 mg. potassium bromid, another 100 mg. and the third 500 mg. potassium bromid for each kilo soil, while one pot served as control. On April 23, ten seeds of Phaseolus (Dwarf variety) were sown and on May 25, the plants were reduced to two of equal height in each pot. The plants measured on July 2: Control plants Potassium bromid plants eS >, > a 10 mg per kilo. 100 mg per kilo. 500 mg per kilo. cm. cm. cm. cm. 22 22 22 22 21 24 24 19 1 Cf. O. Loew, Ein natiirliches System der Giftwirkungen, p. 108. ‘ [aa 2 Journ. Roy. Agr. Soc. Engl. (III) rr, p. 566. 140 K. Aso. On July 24, the plants were harvested. Control Potassium bromid I, 10 mg 100 mg 500 mg per kilo. per kilo. per kilo: Number of frurts (ripe)e.j).cessess=- 4 i 6 6 is (unripe)esae-seeee 2 O I 2 Weight of pods (air dried) ......... 5.0.2: L1.O1e: 8.8 ¢g. The Number af ripened» sceds2.-2 15 26 a 21 Weight of ripened seeds (air dried) 4.4 g. 5:52: 6.5 g. 5.7 g. These figures leave no doubt that 10 milligram potasium bromid per kilo soil had exerted a stimulating action and further that this beneficial action decreased with the increase of that salt. But even 500 mg. potassium bromid per kilo soil had still a slight stimulating action.1 Another experiment was made with upland rice. The manuring and the quantity of potassium bromid were quite the same as in the case with Phaseolus. On April 23, ten seeds of upland rice were sown into each pot and the young plants reduced to seven of equal size on June 4. Later on two more were removed from each pot on account of damage by fungi. The height on July 24 was: Control Potassium bromid aaa a 1omg per kilo. toomg perkilo. 500 mg per kilo. cm. cm. cm. cm. 60 SO 69 68 68 82 74 68 So 74 69 72 67 71 60 64 67 78 7O 58 Average length. 66 77 68 68 1 The seeds of these plants yielded again normal plants. ee Can Potassium Bromid Exert any Stimulating Action on Plants? [41 On October 5, the plants were harvested. Control Potassium bromid LN — eee 10 mg 100 mg 500 mg per kilo. per kilo. per kilo Peverase lenoth ........1-:: 81.4 cm. 87.6 cm. $3.6 cm. 75.0 cm. Weight of grains (air dry). 10.5 g. ns &. 8.7 g. 8.0 g. Weight of straw (air dry). 18.5 g. 222 S, 18.2 ¢g 14.5 g. This result shows, like in the former case, that 10 milligram potassium bromid per -kilo soil exerts a stimulating action but this action diminishes with the increase of the amount; 100 milligram potassium bromid per kilo soil depressed the weight of grains and 500 milligram potassium bromid injured the normal growth of the plants. A further experiment was made with fungi. The culture solutions contained: 0.5 % pepton 1.0 % glycerol 0.2 % monopotassium phosphate 0.02% magnesium sulphate. Potassium bromid was added at the rate of 0.199, 0.019% and 0.001%. These solutions were infected with a trace of spores of Aspergillus Orysae and kept in diffused day-light at the ordinary temperature. For control the solutions received equal amounts of potassium chlorid. After four days, the fungus mass was collected on weighed filters and dried. The result was as follows: Weight of the fungus mass. Seer rN at Wee ave cts ce Nats sss 0.013 ¢. 2h ig met Stem eee ae GOES »; ee cece ead me ae « eee: 4 PAS) ass It will be scen that the stimulating effect of Zn was nil and those of Co and Ni minute and uncertain. In all the 4 cases here described I have observed that in the early period of develompment the nickel plants showed the most favorable growth and from the middle period of development cobalt plants gained headway while the control plants continued its growing and flowering for a longer time than the others. As a general result, however, it may be inferred that stimulant actions can be exerted in certain cases by small doses of zinc, nickel and cobalt salts, on agricultural plants, but this effect was not consider- able in my experiments. Can Lithium and Cesium Salts Exert any Stimulant Action on Phenogams ? BY Mi. Nakamura. Lithium belongs to those elements which are present in very small quantities in many soils.!. It was found also in many plant ashes by Bunsen and Kirchhoff.2, Lippmann found it in the ash of the sugar beet, 7ruchot in the ash of tobacco (0.44%). Zschermak observed that lithium especially accumulates in the leaves, and that certain plant species take up lithium salts much more easily than others. He further denies any relation between the lithium contents and the degree of development. MNodde (1871) had observed that lithium cannot replace potassium in the plants, and that lithium salts act even poisonously upon buckwheat.* With alge and fungi no injurious action of lithium salts was observed.4 It remained now to be seen whether lithium salts are capable. to exert a stimulating action on phznogams when applied in very small doses.* This question seemed to me of sufficient interest to justify some experiments in this direction. 1 Truchot C. r. 78. 1022. 2 Ann, Chem, Pharm. Vol. 118 p. 353. 8 The culture solutions of Nobbe contained 104.2 milligr, Li,O per liter, in another case 73.8 milligr, 4 O, Loew, Ein natiirliches System der Giftwirkungen. p. 115, Also Journal prakt, Chem. Bd, 36, S. 284 (1887). 8 According to Ove lithium nitrate can act as a moderate stimulant of growth on algae and fungi (Journal College of Science, Tokyo, vol. 13 p. 165(1900) ):—Thus Li NO, in a dilution of yk x 107¢ in- creased the growth of Frofecoccus in 24 days, from 0.010 g. in the control case to 0.020 g, and the growth of Aspergil/us niger was stimulated by iy x 107? from 0.300 g. in the control flask to 0.408 g. in 17 days. Richards observed stimulant action of 19% Li Cl in the culture solution on fungi (Pringsheims Jahrb. wiss. Bot. Vol. 30, p. 665 ; 1897). 154 M. Nakamura. The soil serving for my experiments came from our College Farm at Komaba and was manured with 1 g. Na NO,, 1g. K,SO,, 0.5 g- KCl, 0,5 g. (NH,), SO, and 1,2 g. double superphosphate per kilo. To one pot was added 10 milligr. lithium carbonate per kilo soil, to the second 100 milligr. per kilo while the third served for the control plants. The pots contained 8 kilo soil each and were kept in the greenhouse. /“¢ Experiment with Barley. Twenty grains of barley were sown January 16 and the young plants reduced later on to five of equal hight in all the pots. At the beginning of May some difference in development was observed and the measurements were as follows: Number of Average length * 2 3 branches 100 mg. Li,CO, per kilo soil 72. 24 Ow ey 3 * PGi iss 24 | Control Taras 22 On May 27 the measurements were as follows: Average length Boose s ‘ S S branches 100 mg. Li CO, per kilo soil 76.0 cm. 27 10) 5 ” » SNE oe 24 Control ph a of Can Lithium and Cesium Salts Exert any Stimulant Action on Phenogams? [ wu war The plants were cut June 13 with the following result : Weight of | Average | Number Total Weight Number | of of | each 100 length of of . weight grains grains | grains | branches branches saci per kilo 104 g. 44.7 &. 887 | 5.28 z. 76.3 cm, 27 ae eee —_ = Sea pos ie Tide 5, ABS. 5s 808 | Bae, Sis. bn 24 Control 97 38.5 | 787 | 4.89 ., 75-3 » 23 Experiment with Pea. The conditions were here essentially the same as in the first experiment. Twenty grains of pea were sown January 28 and the young plants reduced later on to four of equal height in all the pots. In the early period o¢ development the control plants showed the best growth and also showed some flower several days earlier than the lithium plants. Measurements were made on May 5 with the following results: = | r Average length | Number of | Number of of branches | branches flowers | eae 1co mg. Li,CO, per kilo soil 86 cm. | 5 I [Gs 4; os 2 G6. 3°" 4 3 Contro BOD 5. 3 | 4 6 After that time the lithium plants showed a more vigorous development than the control plants. 156 M. Nakamura. The plants were harvested June 15. Air dry plants. Total Weight Number Number Weight of seeds of seeds of branches 100 mg. 65 g. S055 i: 147 5 IO mg. Giie: Sn os 142 4 Control BOs. 20:5) 45 140 4 Both experiments show that /ithium carbonate can exert a slight stimu- lant action. Under the conditions just described an experiment with upland rice was carried out to test the influence of cesium on the development. Pot A re- ceived 10 Milligr. caesium chlorid per kilo soil, Pot B 100 Milligr. while Pot C served as control. Young plants of equal height were selected from the seed bed and planted into the pots a five in each, at the beginning of June. At the time of flowering it was evident that the plants in B exceeded the others as to height. The plants were cut October 2. In pot A no branches were developed in B one branch with yet unripe seed, while in C one branch with ripe seed. The measurements gave the following figures: A B Gg 100 cm. 109 cm. 109 cm. LOD. 1 Cae TOT 4, to." i i Dae 108 ,, LOG 55 LIA 5, Ol as LO5 44, 2 ae 84: >) Average height. 103 _,, 110 5, 99 Can Lithium and Cesium Salts Exert any Stimulant Action on Phenogams? 157 The height of the branch in B was =58cm; in C=75 cm. The total production of ripe seed, weighed in the air dry state was: 7s B ce Ez. 0 ot. 14.6 g. 13-7 g. It can therefore be inferred, that czesium chlorid at the rate of 100 _ mg. per kilo soil had exerted a moderate stimulating effect, especially notice- _ able in the height of the plants. ike J On the Stimulating Effect of lodine and Fluorine Compounds on Agricultural Plants II. BY K. Aso and 8S. Suzuki. In former Bulletins of this College, a field experiment with radish and pot experiments with oats and pea were described which showed the stimulating effect of small doses of sodium fluorid and potassium iodid. We have continued these investigations, upland rice serving now for the test. Fife plots were selected, each of these had ten square meter and received as manure 100 g. double superphosphate, 200 g. am- monium sulfate and 150 g. wood ash, the last mentioned salt being ploughed into the soil one day later than the former. The seed to the amount of 47.5 g. per plot was sown April 30. Plot I received 0.08 g. Na F, II 0.8 g. Na F, III 0.025 g. KI, IV 0.25 g. KI, and V served as control plot. The solutions were applied in high dilution. On June 2, each plot received as top-dressing 50 g. sodium nitrate and 50 g. monopotassium phosphate. The plants on the fluorine plot surpassed gradually the others somewhat in height. A damage done by a typhoon proved to be insignificant. The plants were cut Sept. 8 and left to dry in the glasshouse. The weight was as follows: 1600 K. Aso & 8S. Suzuki: Stimulating Effect of Iodine and Fluorine Compounds ete. y : [ ic Fie Ratio per ha Weight of seeds Weight of straw unhusked, g. air-dry, g. 80 g. Sodium fluorid 2465 1885 800 g. S 3 2090 1800 25 g. Potassium iodid 2300 1900 250§ ” 39 2000 1810 Control 1970 1920 A stimulating effect on the seed production by sodium fluorid and potassium iodid respectively is here evident with the smaller doses of 80 g. sodium fluorid and 25 g. potassium iodid per ha, while the ten times higher doses failed to give a decisive result. On the Treatment of Crops by Stimulating Compounds. Oscar Loew. * At the International Congress of Applied Chemistry in Berlin, June 1903, Professor Gabriel Bertrand from the Institut Pasteuy in Paris read a paper: “Les Engrais Complémentaires” in which was pointed out that, as to mineral manures, almost exclusively compounds of potassa, phosphoric acid and nitrogen are paid attention to, while there exist rarer elements which occur only in exceedingly small amounts in the plants but may never- theless be of a certain physiological signification, leading to an increase of the harvest. Bertrand proposes to call such compounds supplementary manures (engrais complémentaires). It may therefore not be out of place to call attention to the fact that professors and graduates of this College have during the last three years studied the influence of small doses of various compounds upon the growth of plants and yield of crops and published a number of papers on this sub- ject. A short survey of the results obtained may here be in order since some observations are of theoretical interest and some promise even to be- come of practical value. In the first place such elements were considered which occur in small doses widespread in the soil whence they pass into the plants and through them into the animals. These elements are manganese, fluorine and iodine. The manganese content of vegetable and animal organs, t These Bulletins Vol..5, No. 2 and.4; also this number, Recently again Schuetdewind reported from the experimental farm at Lauchstidt that the root crops yieldja maximal harvest only when farmyard manure is applied in addition to mineral manures, Such observations have been made also in regard to tobacco in America, 3 Vol II, p. 16, 4 Report of the Barbados Experiment Station 1901. On the Treatment of Crops by Stimulating Compounds. 173 drochloric acid was found, corresponding to 38 kilo Mn, O, per ha to the depth of 24 cm. It was, mentioned above that also iodine and fluorine compounds might sometimes be used as stimulants in practical agriculture. Some further re- marks are therefore in order. As to iodide of potassium it acts poisonously upon phaenogams in waterculture even in a dilution of 0.002 per cent. Voelker’ observed in a field experiment that a top-dressing at the rate of one half centiweight per acre, corresponding to 62.2 kilo per ha injured wheat and barley. This quantity, however, seems extraordinary great when com- pared with the amount that caused stimulation in our experiments, namely 25 grams per ha! This quantity can be increased to 250 grams per ha without fear of danger, but a further essential increase should be avoided. Hence a dose of 25 g. per ha may be applied for ten consecutive years; if it is taken into consideration, however, that a part is absorbed by the plants and another passes away in the drainage waters, the number of years might be doubled. But after this period a pause of several years should foliow, during which the use of potassium iodid is suspended. The degree of sti- mulation was in some experiments of S. Swsuk¢ considerable! The dose at the rate of 25 gram per ha produced an increase in the weight of radish of 67% compared with that obtained on the control plot and with upland rice an increase of 16% in grains. At the rate of 250 g. per ha the increase in the weight of radish was 31% while there was no increase in seed pro- duction with rice. As to fluorine compounds of potassium or sodium it must be kept in mind that these are in certain respects still more poisonous than potassium or sodium iodid. While the latter exerts a high degree of poisonous action on all plants with an acid cell sap and but a weak one on objects witha neutral cell sap, as e. g. certain algae, those fluorids are very poisonous for all kinds of vegetable objects independent of the reaction of the cell sap or culture. A few observations may here be mentioned. Algae, such as Spirogyra and Mesocarpus are killed within 15 minutes in a 1 per cent solu- 1 Tn pot experiments stimulation was observed at 0.26 and 2.6 milligrams per kilo soil with oats and pea, while at 26 milligrams a depression resulted with rice. 174 Oscar Loew. tion of sodium fluorid, the chlorophyll bands retract their lobes, the nucleus contracts and soon afterwards the cytoplasm recedes from the cellulose wall: In a solution of 0.1% filaments of algae die within 24 hours. Diatoms and monadines become motionless.in a 0.19% solution within 12 minutes. Some monadines recover their power of motion but this is only of short duration, since after 35 minutes no trace of motion reappears. If 1.cc. of a I per cent solution of sodium. fluorid be added to 99 cc. of culture water containing numerous forms of minute organisms, infusoria and diatoms are killed within 2 hours, while some monadines! still were seen alive after this time. At 0.05 per cent sodium fluorid injures the germinating power of seed, while in dilutions of 0.0001 % to o.co1r% it can stimulate growth of phanogams in water culture. . At 0.001% it prevents the action of lactic acid bacilli (Afront) and at 0.15 g. per kilo body weight it proves fatal for animals (Zappe7ner). Attention must here’ be drawn to the socalled Wzborg’s Phosphate which contains fully 1 percent fluorine and is recommended for agricultural purposes. Since phosphates are often applied in large doses as manures, a highly injurious amount of fluorine would soon be accumulated in the soil. This phosphate is manufactured in Sweden by fusing apatite with sodium hydrate and consists chiefly of a sodium—calcium _ silico—phosphate.? Calcium phosphate is now-a-days also frequently added to the food of young hogs in order to promote the formation of bone. There occur, however, in commerce phosphatic preparations containing fluorids which have caused the death of the animals, as Emmerling has shown recently.® The stimulating effects of small doses of sodium fluorid, viz 80—140 g. per ha have been quite considerable in the experiments of K. Aso, who -has observed further that even a dose of 800 g. per ha does not yet act injurious- 1 The monadines are also in other regards of an unusual resistance power which thus far is not satisfactorily explained, 2 IWilborg’s phosphate was recently reported in the Chemiker Zeitung to have the following com- position; P,O,=22%; SiO,=16%; MgO+CaO=35%; Fe, O,=6%; Al, O,=2%; Ky OF Na,O=18%'; Fluorine=1%. * Centralbl, f, Agr. Chem, June 1903. - On the Treatment of Crops by Stimulating Compounds. 175 ly, although the stimulating effect has decreased, or almost vanished, show- ing that a further augmentation of the dose would not be advisable. » ry a wa ‘oyeydins snoursuvu pur SNOdIoJ JO UOIOV CUVe[NWAS 9Yy JopuN syuvid oo9vqo] 9 yf + UA UJ os | [O1}JU07) $$ — fl ma —_—_—- — AX ALVTd TA “TOA TIO): MIO TAT On the Action of Sodium Nitro-prussid upon Plants. BY Rana Bahadur, from Nepal, India. The highly poisonous character of nitro-prussid of sodium for verteb- rate animals was recently demonstrated by Monzes-Diacon and Carquet.} It seemed of some interest to test whether this salt would be also a strong poison for the lowest animal organisms and for the plants. I have observed that infusoria, crustacea and worms were dead after one hour and half in a 0.1% solution of sodium nitroprussid ; only some monadines were still alive. Diatoms also were killed after a short time in that solution. Filaments of JVzte//a were dead after an hour and hal in the 1.% solution. Young barley and buckwheat plants placed in the 0.1% solution were dead after 18 hours, the leaves having completely lost their turgor and being partially dried up. Leaves of radish and mulberry and branches of Polygonum aviculare were also found much affected in that time, the leaves being more or less withered. The leaves of cherry looked more or less brown when held against the light, while the control leaves were perfectly normal. Since, however, the solution of nitro-prussid of sodium decomposes in direct sunlight and more slowly also in diffused daylight, with the pro- duction of prussian blue and prussic acid,? it may be objected that it is the prussic acid formed by this decomposition which killed the organisms. 2 Chem-Centrbl. 1903, p. 510. 2 The characteristic odor of this acid is very soon noticed with these solutions, 178 Rana Bahadur. I have observed that one of the products of decomposition of this salt by sunlight is nitrous acid. Hence the decomposition of nitro prussid of sodium might be represented by the following equation :— Fe (CN), NO Na, + H,O = NOO Na + Na CN-+ Fe(€N)2 eee On account of this decomposition all further experiments were now carried on in darkness. Young barley plants were placed in 0.01%, 0.1%, 1% solutions respectively, and kept in darkness. In the 0.01% solution of sodium nitro-prussid, the plants were not killed even after three days; while the plants in 0.1% solution commenced after twenty two hours to dry up slowly from the tips downwards. (This is quite different from the observa- tion when the barley was kept in daylight, in which latter case the plant completely lost its turgor and dried up within eighteen hours, and finally the plants in 19¢ solution were killed within twenty two hours). Further to a culture water containing numerous lower organisms 0.01% and 0.19% respectively, of sodium nitro-prussid was added and the flasks kept also in darkness. The microscopical examination of the sediment after twenty-three hours showed that diatoms, flagellata and infusoria were still alive, also the ostracodes and small nematodes were seen in their usual motions. Hence it may be concluded that nitro prussid of sodium is in a diluticn of 0.19% not injurious for lower organisms, provided the daylight be excluded and thus the production of prussic acid from the salt be avoided. Some experiments were also made with mould fungi and _ bacteria. In the case of mould fungi, sodium acetate 0.699 was the organic nutrient while ammonium sulphate, monopotassium phosphate, and magnesium sulphate each 0.199, formed the mineral nutrients. In the experiment with bacteria, meat extract served as nutrient. In the former case 1% in the latter 0.59 of nitroprussid were added and the solutions after infection with Penicillium glaucum and Bac. pyocaneus respectively, were placed in darkness. After about one week Penicillium was developed about equally well as in the control case. As to the bacteria, the growth was noticed one day sooner in the control case than in the chief flasks which, however, showed after one week, a luxurient development. On the Action of Sodium Nitro-prussid upon Plants. 179 Conclusion. Nitroprussid of sodium is a comparatively weak poison for lower animal organisms and green plants, and no poison for fungi, provided the daylight is excluded. Daylight decomposes the salt with the production of prussic acid and nitrous acid. Such a decomposition probably takes place also in the higher animal organism which would account for the highly poisonous character of that salt for the vertebrate animals. ——_~ ~0r -—— On the Behavior of Guanidine to Plants. BY I. Kawakita. I. Can guanidine serve as nutrient for fungi? It has been asserted in a paper published some years ago that guanidine may serve as source of carbon in the development of Asper- gillus niger. This seemed however exceedingly improbable since guanidine yields by hydrolysis carbondioxid and ammonia and _ there is no hydrogen atom in direct combination with the carbon. It is most closely related to urea which also is incapable to serve as source of carbon in the nutrition of fungi. Both compounds, however, are good sources of nitrogen for various fungi. In order to test whether really guanidine can serve not merely as source of nitrogen but also as source of carbon, I prepared the following solution : | NTE ETES Sg oa oa Sane ed ae 100 C.c Guanigine bydrochlorid. oi... ... 60... cee. esees i. Si Monopotassium phosphate ...... .............. O.I gr US Sante sh arin stil) FT O.I gr Two flasks containing this solution were after sterilization infected with spores of Aspergillus niger. For comparison a control flask containing glycocoll in place of guanidine was also infected. After 4 weeks there was no trace of development in the guanidine flasks, while in the control flask there was growth. The same result was obtained with spores of Penicillium glaucum. Even Bacillus methylicus, which can utilize such poor nutrients as sodium formiate, refused to grow in the above solutious after they were neutralised. Hence guanidine canf not serve as a source of carbon for fungi. However after addition’ o- 182 I. Kawakita. 0.19 glucose, development of various fungi, as Aspergillus niger and Bacillus methylicus took place readily. 2. Can guanidine serve as source of nitrogen for Phenogams ? Since Sawa! had observed that urea in moderate quantity can exert a poisonous action upon green plants, it was desirable to compare guanidine and biuret in this respect. The following solution was prepared, WABI Ae omrak. meas mie eee. “ashe eee 1000 C.C Calcium sulphate 25): 410. 219305. 2 2eS eee I gr Macnestm sulphate. ... 53.8 35.1 Before entering into the dicussion of these results, we may consider here the proportion of phosphoric acid absorbed by the rice plants from the soil and manures. The chemical analysis of the crops was made by myself in the usual way. On the Influence of Liming upon the Action of Phosphatic Manures. 201 a4 Phosphoric Phosphoric Phosphoric acid absorbed sco acid in the acid in the ee ee of manures, whole crop, per cent of manures gram gram gram ne al 205 Double surperphosphate ............ 4.167 2.948 2.092 50.2 Shimekasu (Sardine)...........00000+ 53 2.308 1.452 34-5 By J 50 ee Feotineenae ; 1.345 0.585 14.0 mhimekasa (FIErring) ..J.....c.cces6 9 2.483 1.627 39:0 ” SEGAL OD. foes saduneceunceves 3 1.100 0.340 8.2 Arakasa (Fish bone) .....c.00cccccses ” 2.709 1.853 44-5 5 FREAD oe ckcdoccuineasavescs = 1.764 1.004 24.1 Steamed bone meal ........... 0000 “p 2.469 1.613 38.7 z TOME O° 0 Pe + 1.070 0.310 7-4 NCP EAIR eee. iy cesses acru ston secseacet > 1.660 0 804 | 19.3 PE COLO rane otisackcancetses 5s 1.175 0.415 10.0 IVABE'CARG Senioa de ctececisdapas 'a-usased 9 1.418 0 562 13-5 Soames Ge CQO cet ckavadterseseet 3 0.866 0.106 2.6 Besa ANGI CANE: 5 y¢Ceiienis cevdicagexess 59 1.739 0.883 2 » of CaO ica eseecee 35 1.060 0.300 7-2 SOY SDEAMICAKGhalicsescecscetestseeastae " 1.389 0.533 12.8 Piidhaste Rndg eta ON sac cies edness . 0.866 0.106 2.8 PO tergastpenciectwc bevucta jade. oO 0.856 Oo | Oo (oa RCE 0 Ne fe) 0.760 fe) o Again, when the above number for the percentage of double super- phosphate, that is 50.2, assumed to be 100, the relative availability of P, O; in the other manures will be found as follows :— 202 M. Nagaoka. Relative availability of phosphoric acid Kind of manures Without Ca O With Ca O Double superphosphate ..:.......0.0c..eccseees shimekasw) (Sardine). .2.. 5: ss.cober case see cena: 28.7 Shimekasu (Herring) suadacoddoaseboaagsodnonoec 16.3 Arakase (Ash bove) 2 Jcs.isate- covdeeecc seco: 48.0 teamed DOME Med <77.cscersesipecscrednusescdee 14.7 Rice ra 7 2ites - ee SCCAMUM CAKE. ee acdicssasaencctecnesisse 14.0 28.0 21.0 3 | 3% MEM CAINE KCN. v caclcsscacie isewsisdosdseceie 18.7 20.3 19.5 v PePAVET AS Clee Siena s Seve sec ciscseacvencecteereusses 18.1 22.1 20.1 These numbers show, with but a few exceptions, namely for Shimekasu (Herring) and sesamum cake, a singular coincidence in the relative diminu- tion on the yield and the relative decrease of the availability of phosphoric acid; and this coincidence appears still more striking in the averages. The average of these two relative series supplies a quotient of the retarding action of lime upon the phosphoric acid contained in various organic manures, suited for the rice plant under the condition similar to that of our experi- mental field in regard to soil and climate. Now, when a comparison is made in regard to the above numbers, it is observed, in a general sense, that the consumption of phosphoric acid and the yield of the rice plant supplied with manures of animal origin, were affected to an extensive degree on account of lime application, while in the case of vegetable manures, the depressing action of lime was not so great. The respective averages of the diminution quotients of both kinds of manures, that is 50.2 for the animal manures and 20,1 for the vegetable manure, show the relative difference between these two manures ; and these two numbers will be practically of a great value in regard to a paddy field rich in line or over limed, especially to a field which is freshly manured with lime in a large dose. Now what is the cause which brought on this great difference in the re- sults of these two kinds of manures? This may to a certain extent depend upon the relative amount of organic matter in the manure and the relative degree of its decomposition.1_ The humus exerts an important influence upon the availability of phosphoric acid from soils and manures. Thus it has been 1 The opinions as to the action of lime on humus and humification process differ, A essowitsch (1902) infers from his experiments that lime retards the humus formation. 204 M. Nagaoka. demonstrated in the application of basic slag or other phosphatic manures rich in lime that the result was more satisfactory in the presence of much humus than in its absence, what it probably due to the formation of organic acids formed in the humus. By the application of lime the original humus in my experimental soil had been rendered inactive. This fact was well proved by the unfavorable condition of the rice plants in the limed frames which had not recieved phosphoric acid, when a com- parison was made with that of the control frames. In all the limed frames, this deficiency or want of humus action in the soil is counteracted to a certain extent by the application of the above men- tioned organic manures and the notable fluctuation, which was brought upon the harvest as well as the rate of phosphoric acid consumption, is duc chiefly to the quantity and nature of the organic matter contained in the manures. The manures of animal origin containing less organic matter and being naturally more readily decomposed, their humification in the soil was not sufficient enough to bring the insoluble phosphoric acid into a more available state. The acidity of the rice roots which exerts a solvent action upon the phosphates also seems to have been weakened by the liming. And finally the ratio between the amounts of lime and magnesia in the soil may have become unfavorable to rice by the liming, although the amount of lime was not large enough to exert in this regard much influence. The above discussion will receive some further support by the results of the after-effects of this experiment. With special reference to the steamed bone meal, I may quote here the results obtained by Kel/mey and Béttcher. These authors obtained on an average of ten trials with different samples of bone meal a relative retarding quotient of 47 with rye,! and of 50 in the next experiments with white mustard? while in the experiment of the writer the retarding quotient was as high as 54.8. It is very satisfactory to say that without considering the ' Deutsch, Landw, Presse, 1900, No, 52 and 1901, No, 23-24. 2 Dentsch, Landw, Presse, 1901. No, 28. On the Influence of Liming upon the Action of Phosphatic Manures. 205 great difference in climate, in the condition of soil and in the character of the crops there is a very close resemblance between their results and mine. Nevertheless when a strict comparison is made between both results, it will be seen that in my own, the action of lime was more powerful what may most probably be due to the fact that Kel/ner and Béttcher used calcium carbonate while in the experiments of the writer caustic lime had served. ! It may be of some interest to add here the relative manurial effects of the phosporic acid of the various manures most commonly used in the paddy fields of Japan. As to these determinations compare the Bul. Vol. I of this College. In the following table the recompiled numbers from the above pages are added :— Relative action on the plus yield Relative Relative Kind of manures, of total crop assimilibility | manuring over the frame ; without P,O,. oo ee LEU Double superphosphate ...............06. 100 | 100 Shimekasu (Sardine) ......ccceeseseeeeee 68.7 85 n oO 5 Shimekasu (ETenning) i paseckerectecteess 77.5 82 q Gea Atakecu (Bish Bone) »” ” 234.0 Bel, 54 9» ” 299.0 5.0 17 ” ’ Tha) ak Ca O 96.0 2.8 36 ” ” ” ” 152.0 5.1 55 ” 9” 99 147.0 5:0 18 No P,O, 148.0 5.1 37 ” 163.0 4.2 56 y: 155.0 45 19 » » +CaO 140.0 35 38 ” ” ” ” 128.0 33 57 ” ” ”» » 128.0 3-7 On the Influence of Liming npon the Action of Phosphatic Manures. 213 YIELD OF THE SINGLE FRAMES. (2nd year.) Straw, Full grain, Empty grain, No of frames, Kind of manures. grams, grams. __ grams, I Double superphosphate 5.0 20 5 a 4.6 5g ” ” 4.6 2 Shimekasu (Sardine) 4.6 21 3 3.0 40 - 4.8 fe is +CaO 7.3 22 i 1S 6.9 41 # i6:1 35 5.0 4 Shimekasu (Herring) 4.4 23 ” 4.9 42 9 .2 5 ” +Ca O 8.5 24 ys 9 6.2 43 ” 9 6.9 6 Arakasu (fish bone) 4.4 25 ” 5-9 44 »” 2 9 » +020 5-3 26 » ”» 4.6 45 ” 9 4.1 8 Steamed bone meal 10.1 27 ” ” ” 3.5 46 9 ” 9 3-5 9 99 » +CaO 5.6 28 ts % 9 Seb 361.5 | 268.6 6.7 ” ” » 99 263.0 | 200.6 | 4.2 ae Cae eaaes 214 M. Nagaoka. YIELD OF THE SINGLE FRAMES. (2nd year.) Full grain, | Empty grain, No of frames. Kind of manures. grams, grams, 10 Rice bran 152.7 3.4 29 AAs 190.1 133.7 BAS 48 a es 234.1 117.1 3.0 II + » +CaO 254.8 167.6 6.1 30 ” 999s 236.0 170.6 3.7 49 7 set a3) Gs 235-5 162.6 4.6 12 Rape cake 201.0 141.6 3.6 31 sa. 435 240.0 161.6 4.3 50 Sys 244.0 180.6 333 1 so ree 291.0 198.6 5.7 32 99. 99998 314.0 222.6 4.8 51 be kor a. 304.0 225.6 4.7 14 Sesamum cake 241.5 156.1 4.9 33 a ‘3 310.0 147.1 2.8 52 ip Fe 236.5 187.6 3.9 15 ” » +CaO 264.0 179.1 4.9 34 a Se ae 253.0 178.6 5.5 53 se rel eee 299.0 206.6 6.3 16 Soy bean cake 178.0 114.1 3.6 35 a es i 258.0 194.6 4.7 54 the ee 5 218.0 156.6 3a 17 » » » Cad 295.0 223.1 6.6 36 gp VASO idige 265.0 199.1 5.4 55 ay. ss re: rey 269.0 204.1 4.4 18 No P,O, 188.0 134.6 a 37 oo” 170.0 115.6 2.9 56 » oo» 182.0 125.6 2.9 19 » » *+CaO 240,0 160.6 3.9 35 » 9 9” 245.0 161.1 4.2 57 share oe 242.0 163.6 4.8 On the Action of Various Insoluble Phosphates upon Rice Plants. BY M. Nagaoka. The phosphoric acid of the soil occurs partly in a soluble form while the larger part is present as compounds insoluble in water. Among the insoluble forms the phosphates of iron, alminium and calcium predominate. Besides these mineral forms some phosphoric acid exists also in form of an organic compound, thus far not very closely studied. As to the origin of the insoluble mineral phosphates, there are two sources, one being the mineral particles of rocks which always contain more or less phosphate of calcium, aluminium and iron in the form of phosphorite, vivianite and wavellite respectively, the other source being the soluble phosphoric acid in the manure, being transformed in the soil to the aluminium, iron and calcium phosphate. Since the iron and aluminium phosphate are insoluble in dilute acids their manurial effects upon crops thus far observed are but of small value. Since, however, the paddy fields for the cultivation of rice are irrigated for the greater part of the year, their soil conditions are entirely different from those of the dry fields. Among these conditions, the feature of the decom- position of organic matter is most noteworthy. Thus the decomposition or putrefaction of organic matter easily takes place in paddy fields, with the result that an acid humus is produced, since the oxygen does not penetrate sufficiently into the wet soil to lead to a complete oxidation. Paddy fields have a more or less acid nature according to the amount of humus present. This acid reaction must naturally have a beneficial action, since the insoluble phosphates become thus more easily available for the roots. This led me to carry out some experiments with the intention of obtaining some informations on the effect of various insoluble phosphates upon rice plants. 216 M. Nagaoka. I. Series. These experiments were performed in wooden frames (3 square shaku each) which were, after the usual manner, placed in an uniformly agitated and levelled paddy field, the soil of the latter having been exhausted for four years by the continuous cultivation of rice crops without manures. After the careful management of the soil in each frame, general manures were given. Thus, on the 23rd of June 1898, the potassium sulphate was added at the rate of 100 kilograms of potash per hectare and four days afterwards, the same rate of nitrogen was applied in the form of ammonium sulphate. On the next day, the special phosphates were given. The quantity of the phosphoric acid employed and the amount of the corresponding phosphates will be seen in the following table. Quantity of phosphoric acid, | Quantity of the phosphates No of frames. Kind of phosphates, used per per hectare per frame frame, kilograms. grams. grams. E5040; 17.9! No phosphoric acid. oO fo) fe) Ay Aap iis2 25 2.083 8.711 55 144, 183: Ferric phosphate. 50 4.166 17.422 6, 45, 34 34.844 13, 52, 91 7-299 14, 53. 92 14.598 15, 54, 93 29.106 22, 61, 100 6.0018 Aluminium phosphate. 50 4.166 12.002 24.004; 5.039 Calcium phosphate. On the Action of Various Insoluble Phosphates upon Rice Plants. 217 The above quantities of the four phosphates were calculated from the following analytical results :— In percent of the air dry samples. ig gbettic- phosphates <= yee eA; 723 a i Merrousiphospiate: .2. 4.. oi: -. . 345825 3. | Aluminiim.phosphate <«.. ... ... 52,001 a> jCalcigml phosphate ... (/.. .... ..--..50,123 With the exception of the calcium phosphate, these numbers show close coincidence with their theoretical content in phosphoric acid. The calcium phosphate applied was guaranteed to be a pure tricalcium phosphate ; but according to the above analysis it showed a somewhat higher content of phosphoric acid, making it probable that the sample contained some dicalcium phosphate. On the 2oth of June, the young rice specially grown for the purpose and 52 days old was transplanted to each frame. Each frame received 16 bundles of twelve healthy individuals. The irrigation was at once commenced in the usual manner, but was discontinued at the end of August. However, according to the system of the common practice of rice culture, the frames were again irrigated for three days during the full bloom of the rice plants. Refering to the report of the meteorological observatory of Tokio, the weather, during the whole vegetation of the rice plants, showed fewer clear days than usual, but temperature and rainfall were as favourable as in other good rice years. On the zoth of November, the crops were harvested and exposed to the sun for about ten days. Thus, the crops having been completely .air dried, the grains were carefully isolated from the panicles. The average yields! of three equally treated frames were as follows :— 1 The yield per frame is left to the appendix. 218 M. Nagaoka. PO; Average yield per frame, applied Soy Full E No of frames, Kind of phosphates. | per ha, Straw, e mpty Total grain, grain, crop, Kilograms,| grams. grams, grams. grams, Teo, 70 341.0 289.3 7.4 638.0 4, 43, 82 488.0 451.3 r2i7 952.0 e445 Oss Ferric phosphate 668.0 573-5 23.2 1265.0 ONL Ab tod: 816.7 636.0 20.7 1473.0 I Sey Or 476.3 408.6 12.8 898.0 145 a Oe: Ferrous phosphate 552.3 458.6 13.6 1025.0 ie Gey Ce 632.3 529.7 19.1 1171.0 26, 61, 100 457-0 366.7 12.0 836.0 27a O2; LOLs Aluminium phosphate 570.0 465.3 14.2 1050.0 25> (63 Oz: 649.7 554-7 18.6 1223.0 31, 70, 109 500.0 446.7 II.0 958.0 32, 91, 110. Calcium phosphate 618.3 495.7 18.8 1132.0 oa 92) OLNTs 715.3 588.7 22.8 1327.0 It will be seen from the above figures that all the series of experiments showed a prominent influence of the phosphoric acid of the various insoluble phosphates upon the yield of the grain, as wellas of the straw of the rice plants, and in every case it will be further seen, that the greater the amount of the phosphoric acid was, the larger was the yield. The increasing rate upon the yield by the action of different doses of the various phosphates is still more clearly explained in the following calculations. Thus, when the plus yield of the smallest doses of the respective phosphates over the frame not supplied with phosphoric acid, is assumed to be 100, the other plus yields will have the following ratios. On the Action of Various Insoluble Phosphates upon Rice Plants. 219 Phosphoric acid Plus yield over the Relative increase lied frame not supplied Kind of phosphates. so ei Ain with phosphoric SE, ha, acid, different doses. kilograms, grams, 314 Ferric phosphate Ferrous phosphate Aluminium phosphate Calcium phosphate It will be observed from the above figures that the relative increase caused by the medium doses of the ferric and aluminium phosphates is nearly the same, while the increase by ferrous phosphate and calcium phosphate showed also a close coincidence, thus the medium doses of the former phosphates increased equally the yield 100% more over the yield of the smaller doses and the latter phosphates gave likewise almost 509% more. The relative increasing yields of the largest doses of the respective phosphates were not insignificant and there is, excepting the ferric phosphate, still almost the same climax of the yield over that of the medium doses, as that which between the smallest and medium doses. In fact, the crop produced by the largest amount of the phosphoric acid was considerably higher than that obtained with the smaller quantity, and this proves that the smaller and the medium doses of the phosphoric acid displayed throughout all trials their full actions and that the increase obtained with the medium doses over the frame without phosphoric acid, 220 M. Nagaoka. gives a reliable measure of the relative value of the various phosphates applied. In order to compare the effects of the various insoluble phosphates with that of soluble phosphoric acid, a special trial was made with double superphosphate on a part of the same field in a similar manner; 5 kilograms of the phosphoric acid of the double superphosphate gave on average 581 crams of straw, 491 grams of full grain and 13 grams of empty grains which makes 1085 grams of total yield per frame. Taking, now, the plus yield of the double superphosphate over the frame without phosphoric acid to be as 100, the relation of the increase with the application of 5 kilograms of the phosphoric acid of each phosphate is calculated with the following figures :— Surplus over the frame Relation of the increase. Kind of phosphates. without phosphoric acid. Surplus of the dov ole super- grams. phosphate = 100 Double superphosphate ............. 100 Ferric puOsplate.vcssccurscxavesusoses 140 Ferrous phosphate,........ssccsesseres 87 Aluminium phosphate,............+6 92 Caleiam. phosphate,.....seocreeeose 117 Before entering into a discussion on these results we may further consider the proportions of the phosphoric acid consumed by the rice plants from the soil and the respective phosphate, as found by the chemical analysis made by myself. On the Action of Various Insoluble Phosphates upon Rice Plants. 221 P,O, in the P,O, consumed from #20; pphed: the phosphate. Kind of phosphates. total crop, Per ha. Per frame % of the kilograms, grams. grams, grams. P.O, applied. Without phosphoric acid fo) o 1.369 — —- Double superphospha'e 50 4.166 2.812 1.443 34.6 Ferric phosphate 25 2.083 2.170 0.801 38.5 ” : 50 4.166 2.930 1.561 37-5 ” » 100 8.333 3.953 2.584 31.0 Ferrous phosphate 25 2.083 1.947 0.578 277 ss ns 50 4.166 2.301 0.932 22.4 ” 9 100 8.333 2.705 1,336 16.0 Aluminium phosphate 25 2.083 1.761 0.392 18.8 7A we 50 4.166 2.138 0.769 18.5 ” ” 100 8.333 2.557 1.188 14.3 Calcium phosphate 25 2.083 2.026 0.657 31-5 ” ” 50 4.166 2.459 1.090 26.3 ” ” 100 8.333 3.083 1.614 19.4 It is thus seen in all trials, that from the larger dose of the phosphoric acid applied, considerably more of this phosphoric acid was consumed by the rice plants than from the smaller dose; and it will be most reasonable to say that the results obtained with the medium doses constitute again a reliable information on the availability of the phosphoric acid of the various insoluble phosphates. Assuming, now, the availability of the double superphosphate (34.6) to be roo, and calculating, on this basis, the relative availability of the other forms of phosphoric acid, the following numbers are obtained, and when a average is made of these numbers as to the relative action on the increase of total yield, the results will express the manurial value of the various phosphates for the first year. 222 M. Nagaoka. Relative Relative action on Relative manurial Kind of phosphates, the increase of value in the first assimilability. total crop. year. Double superphosphate ...... 100 100 Fetric phosphate) -<-..secc 108.4 124 Ferrous phosphate ............ 64.8 76 Aluminium phosphate.,....... 53-5 73 Calcium phosphate ............ 76.1 97 As all the precipitated phosphates are naturally insoluble in water and as they are not easily distributed through the soil, I did not expect such a considerable good effect and rapid influence upon rice plants, and the results obtained by the experiments of the first year sufficiently prove that the insoluble phosphates which have been hitherto believed to be of very low value, have a fair action upon the rice plants cultivated in a soil of a certain character. The condition which brought out these unexpected good results in the present experiment, is most probably due to the presence of much acid humus in our soil, the acidity of which having played a most important rdle upon the solution of the insoluble phosphates. The further discussion on this point is left for the later pages. Tl. Series. After-effects of the insoluble phosphates. (Second year). In order to determine the further action and the value of the various phosphates, the series of the preceeding experiments were continued for the second year (1899). All the frames after the harvest of the first crop, were left untouched till the end of May, but the soils in the frames were exposed to the air during the whole winter in a sufficiently dry state. On the 20th of June, after the soil had been previously ploughed, all the frames equally received nitrogen and potash respectively at the rate On the Action of Various Insoluble Phosphates upon Rice Plants. of 100 kilograms per hectare and in the form of sulphate. 22 <<) On the 25th, the young rice plants of the same variety as of the previous year were transplanted in the same manner; also the treatments of the plants was not essentially different. No parasites were observed. The weather was most favourable for rice plants. On the 21st of November, the crops were harvested with the following yields ; the numbers are the average of three equally treated frames :— No. of frames. Iol. 102. 109. 71, 72, IIo. 33» III. Specially tried Kind of phosphates, Without phosphoric acid, Ferric phosphate Double super- phosphate P.O, P.O, left applied | from the in the first year| ing crop. per. ha. |per frame. preceed- Average yield per frame. Straw. grams. —_—. | | hp» Emply | Total grain. crop. grams. | grams, 5:5 512 5:7 592 6.2 637 7.0 840 59 547 5-5 So 6.4 754 6.2 522 6.2 641 6.8 697 6.3 551 7.6 610 5.8 628 6.5 601 In all the experiments, it was observed that the unrecovered phosphoric acid had an influence to a certain extent on the second crop. However, there was no case that the crops of the second year exceeded that of the first 224 M. Nagaoka. year. Further, the greater was the amount of phosphoric acid left from the preceeding year, the larger was the harvest. In order to arrive at a definite idea, as to the influence upon the increase of the tolal crops, I calculated from the above results of the medium doses, assuming the plus yield of the double superphosphate over the frame not supplied with phosphoric acid to be 100, the following numbers; and for the sake of comparison, I add here the results of the first year. ee ee eee —————————————§ Plus yield over the frame Relation of the increase without P,O,, surplus of the double : hosphate = 100. Kind of phosphates. ae i i eae ist year 2nd year 1st year 2nd year Double superphosphate ............... 447 &9 100 100 Ferric phosphate. ..........0++.0+seeeen 626 125 140 I4I Ferrous phosphate ...............-0+6+- 387 78 87 88 Aluminium phosphate .. ............ 412 129 92 145 Calcium phosphate...............00066 495 89 117 110 The two series of figures on the relative increase coincide tolerably well with the one exception of that obtained by the aluminium phosphate which seemed to be relatively more effective, owing to its late and gradual decomposition in the soil. Before entering into a full discussion of the above results, I may here consider the amount of the phosphoric acid absorbed by the plants from the various phosphates in the second year. The chemical analysis of the crops of the second year gave the following figures :— On the Action of Various Insoluble Phosphates upon Rice Plants. 225 Consumed from the P,O, applied} Unrecovered | P,O, in the unrecovered P,O,. in the first POs total crop : year. Per frame, of second Kind of phosphates. Pen thiay year, Widen ca 0% of the unrecoverd kilograms. grams. grams, grams. P50: Without phosphoric acid, fo) — 1.105 — — Double superphosphate 50 2.723 1.406 0.301 11.1 Ferric phosphate 25 1.282 1.308 0.203 15.8 » ” 50 2.605 1.520 0.415 15.6 ” ” 100 5-749 1.955 oO 850 14.8 Ferrous phosphate 25 1.505 1e2e7, 0.132 8.8 7 iS 50 3.234 1.362 0.257 7-9 ” 9 100 6.997 1.816 0.711 11.6 Aluminium phosphate 25 1.691 1.257 0.152 9.0 ” ” 50 3.397 1.470 0.375 ce ” 9 100 7-145 1.638 0.533 7.5 Calcium phosphate 25 1.426 1.260 0.155 12.5 ” ” 50 3.076 1.349 0.244 7:9 » 100 6.719 1.439 0.334 5.0 Assuming the availability of the residual phosphoric acid of the double superphosphate (11.19%) to be 100, the relative assimilability of the other kinds of phosphates will be as follows :— Relative assimilability. Kind of phosphate. In the first year. In the second year. Double superphosphate .................. 100 100 BRGEEIG PMOSBEGLS . ci o.ccuvccvcvevecedensccns 108 | 142 PECEOUS PNOSPHAtE .5. oo... cesseceneave 65 | 71 Aluminium phosphate ..............0..0... 54 | 99 Calcium phosphate .............s.ccceseees 76 | 7! 226 M. Nagaoka. Thus it is seen, that the rates of the relative assimilability in the second year are, but with the exception of the calcium phosphate, greater than that of the first year in every trial, and this may be due to the nature of the gradual efficacy of the various insoluble phosphates. As in the first year, I calculate the manurial value of the various phosphates for the second year, making an average of the two series of results, that is the relative increase on the yield and the relative as- similability of the residual phosphoric acid. Relative manurial value. Kind of phosphate. ———————————— Average. Ist year. Second year. Double superphosphate ... ........... 120 100 100 Berrie phosphatenwns secs pe cesedaets 124.2 141.5 133 Ferrous) phosphate’ <-22-2-.+c-useneaees 75-9 79.5 78 Aluminium phosphate.................. 72.8 122.0 97 Calcium phosphate «7... -:scpvsaiess00 96.6 90.5 94 Thus two years experiments have shown a most reliable relative value of the various insoluble phosphates. The best effect was displayed by the ferric phosphate in both years; however this phosphate being the most insoluble among all the phosphates experimented on, I did not anticipate such an extraordinary result. The real cause for this phenomenon is left to future researches. (Acidity of humus ?) Next to the ferric phosphate, the calcium phosphate has acted pretty well in the first year, but its action was somewhat decreased in the second season. The ferrous phosphate and aluminium phosphate had less value in the first year, but the latter phosphate has shown a prominent action compared to the other phosphates in the second year; still the yield of the second year did not exceed that of the first year. On the Action of Various Insoluble Phosphates upon Rice Plants. 227 LIES es: The after-cffect of the residual phosphoric acid in the 3rd and 4th year. (1900 and 1901). The former series of the experiments have been carried on as far as the 4th year in order to estimate the action of the residual phosphoric acid left by the successive rice crops. In each year, the frames were treated just in the same manner as in the second year, and every year they received, in the middle of June, only nitrogen and potash as general manures, in the form of sulphate and at the rate of 100 kilograms per hectare respectively. In 1900, the crops were harvested on November 20 and in 1got, the 4th crops on November 23. The average yields of the three equally managed frames are, for both years, as follows :— THE YIELD OF THE 3rv YEAR (1900). Ss : : S reas Average yield per frames, | urplus Fae | over the ae, ip ; ~»1 Uo | fram No of frames, Kind of phosphates, © = ==! Straw Full | Empty | Total prea ons) grain grain crop | po. eS grams. | grams. | grams. | grams. | att 4 > : > = - > grams, ta) 40:8) 70 | Without phosphoric acid fo) 278.0 242.0 2.8 522.8 | : | | 4, 43, 82 | Ferric phosphate 25 281.0 243-5 | 3-4 527-55 | 4.7 Bot 445. 383 bs 50 340.0 287.7 2.5 631.2 | 108.4 = - s = a Gr 455. O44, 100 390.6 360.0 3.4 754.0 231.2 egy Ses OL | Ferrous phosphate 25 286.7 237.0 4.0 527.7 | 4.9 14, 53, 92 | ” ” 50 312.3 260.1 3-2 575-6 ive52 5 | Ws he ates: fgee » 1co 308.6 DEAS A. 3.9 580.5 | 57.3 = | 22, 61, 100 | Aluminium phosphate 25 340.4 BAST ott AG 623.1 100.3 | = | ~ | -o Bee O25) 101 5 - a eOu. (SS oie BOL.G, |}. 1.4.0.) 062% 139.9 24, 63, 102 | ) - | 100 372.3 313.8 4.1 | 690.2 167.4 305, \/0, 109 Calcium phosphate 25 288.8 2318 | 28 523-4 0.6 Ree Ty TO 7 » | .§0 314.5 248.3 | 3.8 | 566.6 43.8 Sih Via » ~ joo: | 376.2 297.3 | 4.1 677.6 | s4.8 e 4 Specially tried | Double superphosphate 50 | 310.0 237.0 | 3.0 | 550.0 | 27-2 228 M. Nagaoka. THE YIELD OF THE 4tn YEAR (r90r)- ] 2. Average yield per frames, Surplus eter over the eee : ‘es : ie Reo e z é F frame No of frames, Kind of phosphates. “a se | Straw Full | Empty Total) aate oF-5 grain | grain crop P.O. a = | grams. | grams. /} grams. | grams, grams, | I, 40, 79 | Without phosphoric acid o | 188.3 G7e8) aiaeeeas 358.6 | | — | ANAS Oe Ferric phosphate | 25 214.0 167.7 2.4 384.1 25.5 | | ee 7 ee oe) # zs, he iGo 246.0 | 198.2 2.8 447.0 88.4 j | Great wot a rs 100 336.3 283.7 e7/ 623.7 265.1 13, 52, 91 | Ferrous phosphate | 25 | 229,0 | 175.0 | 2.8 400.8 48.2 145.5535 492) | = 93 50 235.0 | 186.7 | 3.6 425.3 66.6 } 15) 54 93 | + 100 | 275.0 | 233.2 | "3.4. Sy Sue nnery ae | 22, 61, 100 | Aluminium phosphate a5 | 250.0 | 205.5 | 3.0 458.5 99.9 23, .62, 101 . - | “50 285.0 | 240.0 |: 3.7 528.7. |) ¥7Our | | | 24, 63, 102 i x 100 auike/ 270.3 )0ei | eeoeD, 585.2 226.6 | a are Fi ies ee 31, 70, 10g | Calcium phosphate | 25 265.0 206.0 3.0 474.0 | 115.4 Sore STON As 6 150 292.3 Pe ghey Gal | ees 538.9 | 1803 | | aoe 2, PUL ¥ %s 100 16.3 | 205.77 We 8 585.3 226.7 { } | SE 2 Ae ee | ee ee — = Specially triel | Double superphosphate 50 285.0 |, 220,05) saa o 511.0 152.4 t | According to the above results, it will be clearly observed that the phosphoric acid applied in 1898 had still a distinct effect on the crops up to three as well as four years, but when the sums of the actual yield of the above table are compared with these of the preceding two years (1st and second years) a gradual decrease was obvious, showing that the phosphoric acid given inthe first year was exhausted to a great extent, or was convert- ed in a difficulty soluble form in the soil. The surplus yields of the 4th year over the frame not supplied with any phosphoric acid, were, in all series, greater than those of the 3rd year and it might seem that the residual phosphoric acid in the 4th year had displayed a better action than in the 3rd year, but when we take into consideration the total yield in the frame without phosphoric acid in both years, it will be clearly noticed that the apparent plus yield of the fourth year is not due to efficacy of residual phosphoric acid but to the On the Action of Various Insoluble Phosphates upon Rice Plants. 229 diminution of harvest in the frame not supplied with phosphoric acid, the latter frame being exhausted of its phosphatic nutriment in the fourth year to a considerable extent. Now assuming the third and fourth year’s plus yield of the double superphosphate to be 100 respectively, the relative increasc caused by the respective medium doses of the other phosphates will be found as follows. First year. | Second year, | Third year. Fourth year. | | be Bi a : ae | Double superphosphate .......,....... | 100 100 100 | 100 A | MeUIC PHOSPHATE... oe... cece ccc ecaees 140 141 399 55 Bemrous PHOSphate 2.22.0... case s- os 87 88 194 44 Aluminium phosphate.................. g2 145 514 112 Calcium phosphate... 6.052065. 5.06. 117 110 161 118 These figures show that the effects of the residual phosphoric acid of the various phosphates in the third and fourth year varie within wide limits. In the third year, the best action was displayed by the aluminium phosphate and ferric phosphate, while in the fourth year, the aluminium and calcium phosphates were best. The remarkable effects of the aluminium aud also ferric phosphates in the third year will most probably be due to the action of humus left from the second year upon the residual phosphoric acid, because, in the second year, both phosphates gave a comparatively higher produce than others and consequently the organic matter, as roots and stubbles, left in the soil, were also larger than in the other frames. IV. Series. The action of caustic lime and calcium carbonate upon the effect of various insoluble phosphates. In 1891, Dr. Kellner! then professor in this college, had observed that 1 Bulletin, College Agr., Imperial University, Tokio Vo!. 1 No. 9. p. 23. 230 M. Nagaoka. basic ferric phosphate treated with lime water in presence of much carbon dioxide is decomposed and more than 14% of the total phosphoric acid passed into the filtrate. From this observation, he inferred that “such a process will, of course, also take place in soils between the basic phos- phates of iron and freshly applied lime, which will display its action there also in both forms, as hydrate and bicarbonate ; as a result the crop will be benefited as to their nutrition with phosphoric acid.” Further he added that similar processes are certainly accomplished still more easily between calcium compounds and ferrous phosphate. In order to confirm practically his observation and to get further informations on the actions of lime compounds towards various insoluble precipitated phosphates, these series of experiments were carried out in conjunction with the preceding experiments. These experiments have also been progressing since 1898 and were still continued up to 1901, in order to study the after-effects of the various phosphates under the influence of lime compounds. The methods in these trials were not essentially different from those followed previously, but the calcium compounds were applied in the first year as early as 14 days before the special phosphates. The calcium compounds used were pure freshly burnt lime and pure precipitated calcium carbonate ; these were applied to the soil at the rate of 4000 kilograms of CaO per hectare, which quantity is the usual dose for most of the Japanese paddy fields. The general plan of these experiments will be seen from the following table :- On the Action of Various Insoluble Phosphates upon Rice Plants. 231 No of frames. Zt 4I, 3, 42, 7; 46, 8, 47; 9% 48, 10, 49, II, 50. r2; iSite 16, 555 17, 56, 18, 57, 19, 58, 20, 59, 2y, 60, 25, 64, 26, 65, aie 66, 28, 67, 29, 68, 30; 69, 34, 73: 35, 74; 36, 75 37; 76, 38, 77; 39, 78, 107. Kind of the phosphate. No phosphoric acid ” ” be Ferric phosphate Aluminium phosphate Calcium phosphate > »” ” ” Phosphoric acid applied per ha, kilograms. | | Kind of lime compounds, With CaO With Ca CO, With Ca O With Ca O Already two weeks after the transplantation of the young rice some distinct differences were observed upon the limed and unlimed soil. All 232 M. Nagaoka. limed plants showed a somewhat unhealthy condition while those not limed appeared throughout vigorous and healthy ; in the course of time these differences decreased to a certain extent, yet at the approach of ripening an unfavorable condition of the limed plants was still distinctly noticeable by the comparatively short and somewhat thin stalks. The crops harvested on the 29th of November, yielded the following numbers. ! 2, Z Average per frame. ee 2Za58 | No of frames. Kind of manures. (Ses S be Straw Full Empty | Total es < | grain | grain crop | i | grams, | grams, | grams. | grams. 2, 41, 80. | No phosphoric acid with Ca O o. | 2940 | 227.5: ae | 527.6 SAA ESTP |S ¥ ig CaCOy 10 aa ere nearss | 5.6 | 588.3 7, 46, 85. | Ferric phosphate with Ca O 25 449.6 | 345.8 | 9.9 805.3 3, Aya) 00: a - °c 5 50 457-6 | 407.0 12:9 | eS7725 9, 48, 87 53 3 + 53 100 639.3 570.7 17.4 1227.4 1o, 49, 88. 9 ss KGrRCO}, 25 445.3 38016) 4 LIco $36.9 ey Tfepp Kel = - > 3; 50 609.0 | 511.6 22 1141.8 123) 51; 9 90 “ + s 5 | 100 | 698.6 | 542.0 2ieT 1261.7 op, ia, Ek Ferrous phosphate with Ca O eee 3300 | 281.7 ree oy 17, 56, 95. 5 : 8 so | 3690 | 3190 | 7.7 |; 16957 18, 57, 96 ” ” ere 100 | 504.7 | 400.0 | 849 913.6 19, 58, 97 » , » CaCO, 25 | 412.7. | 349.0 0.7 771.4 20, 59, 98. ” » ” ” 50 | 539:0 | 445.0 13.3 997-3 21, 60, 99. : ¥ a 7 | 100 | 623.7 | 532.7 r.7° | Treen 25, 64, 103. | Aluminium phosphate with Ca O | 25, 35733 | 286.3 8.7 652.3 26, 65, 104. | ” ” Te hee” 392.3 | 321.7 75 722.5 27; 66, FOG. | 3 3 45 » | 100 | 454.0 403.3 10.4 867.7 28, 67, 106 » 9 » CaCO) 25 410.0 | 450.5 2 867.7 29, 68, 107. | » ” 9 | 50 | 460.0 411.0 9.7 880.7 30, 69, 100. | yo is » » | 100 | 578.0 | 514.7 | 13.9 | 1106.6 34, 73, 112. | Calcium phosphate with Ca O his Zs 408.3 | 312.0 9.2 729.5 355 74, 113. es * ay as 50 446.7 351.0 13-7 | 811.4 36, 75, 14. | , - bs nse | 100 510.0 | 421.0 | 12.5 943-5 37, 76, 115 : > » CaCO, | 25, | 5000 | 43gg |) gunmen 38, 77, 116 ‘i m : BO. 41 485.7 | 1071 6 561.0 1216.0 Before entering into the discussion of these results, I may here calculate the surplus yield over the frames not supplied with phosphoric acid and 1 The yield per frame is left to the appendix, On the Action of Various Insoluble Phosphates upon Rice Plants. 23 ios) lime, and for the sake of comparison the plus yields of the unlimed frames are added in the last column. | Phosphoric Plus yield over the frame not supplied with acid applied phosphoric acid and lime. Kind of phosphates. per ha, i: - | kilograms. With CaO. | With Ca CO,. | Without lime. Without phosphoric acid | fe) Ka) now! (—) 49.7 —_—— vie - | Ferric phosphate 25 | 167.3 | 198.9 314.0 | | ” » | 50 | 239.5 503.8 627.0 | x . | 100 589.4 623.7 $35.0 | ' AAs | Ferrous phosphate 25 (—) 19.2 133.4 260.0 ” 50 57-7 359-3 397.0 5 » 100 275.6 524.1 533-0 s | | | Alaminium phosphate 26 14.3 229.7 198.0 ” 50 | 84.5 242.7 412.0 | 3 = 1co | 220:7 468.6 587.0 Calcium phosphate 25 O1.5 PIS HS) 320.0 ” ” 50 173.4 433-6 495.0 » ” 100 305.5 578.0 689.0 Thus we see that in all trials, excepting the frame without phosphoric acid, and also the smallest dose of the ferrous phosphate, the phosphoric acid applied in the form of the precipitated phosphate had again a remarkable action upon the rice production, but when a close comparison is made to the surplus yield of each unlimed frame, some significant decrease in the respective series was plainly observed with but the exception of the smallest dose of the calcium phosphate with the calcium carbonate which, on the contrary, gave a still higher plus yield than the control unlimed frame. These diminutions were, without doubt, caused by a prompt action of the lime upon the various phosphates and it was further observed that even 234 | M. Nagaoka. in the frames not supplied with any phosphoric acid, the yield diminished to a high extent in consequence of the lime application. In order to clear up more fully the injurious action of the lime, the following calculations were made in which the surplus yield of the unlimed frames are assumed respectively to be 100. | hosphoric acid Relative diminution of the plus yield. Kind of phosphates. | applied per ha, | Kilograms. | With Ca O. With Ca CO,. Ferric phospliates. 02. .2 9 ” | 100 7-276 407.9 317-7 7-3 732-9 19, 58, 97- | ” ” » CaCO, | 25 1.637 315.6 225.4 5.1 540.1 20, 59, 98 | 2 2 a a 50 3.386 | 316.0 | 237.3 5.2 558-5 a, 6,99 | » » » » | roo | 6623 | go71 | 317-7 | 73 | 7324 | Z = free! 25, 64, 103. |Aluminiumphos. withCaO) — 25 1.731 | 3788 | 287.5 7.0 | 6733 26, 65, 104 7 eet OR 50 3.613 | 331-9 | 291.0 7.2 | 630.1 27, 66, 105 ” ” ” ” 100 7.386 446 7 352.5 dS. | $07.3 28, 67, 106. = Mage vO, |)\ oc E713, | 2981 | 215.0 |] 63 5194 29, 68, 107 » Soe Ae: 50 3-463 | 337.6 | 251-7 | 65 | 505-8 30, 69, 108 ” 19999 100 7.106 | 403.4 310.0 | 7.9 | 721.3 34, 73, Li2 Calcium phos. with Ca O 25 1.622 | 323.8 241.3 ie) 571.0 | | 3 35, 74, 113 ’ 9 » 99 50 3-545 357-4 | 254.7 | 6.6 618.7 36, 75, 114 a ‘eae : 100 7.379 | 405.2 | 305.7 76 | 718.5 Bis, 078s 115 » 9 oe Oa 3 9 ee 1.524 323.9 227.3 | 64 | 557.6 | | 38, 77, 116 > i eis : 50 3-310 | 329.3 | 242.0 6.6 | 577.9 | ; 39, 78, 117 5 ees ; 100 6.716 | 371-5 | 281.7 | 78 661.0 | 238 M. Nagaoka. The above figures show that in nearly all the trials, the larger the amount of phosphoric acid, that was left from the preceding crop, the greater was the yield, and that the effect of these residual quantities of phosphates was greater than those of the unlimed frames. In order to demonstrate these points more fully, the actual surplus yield over the frame without phosphoric acid and lime compounds was calculated. Kind of phosphates. No jean Ferric phosphate. Ferrous phosphate, Aluminium phosphate, Calcium phosphate, Surplus yield over the frame Bo eat without phosphoric acid and frst year | Hime compounds. (grams. per ha, with with not kilograms,| CaO. Ca CO,. limed. fe) 84.9 26.1 25 184.4 53.0 50 255.2 167.6 100 387.9 285.7 25 199.4 34.1 50 222.6 46.5 100 220.9 220.1 25 161.3 7.4 50 118.1 83.8 100 205.3 59.0 106.7 206.5 frame = 100, Surplus yield not limed with Ca CQ,. (=), 377 (+) 34.1 (—) 12.9 (-) 2.6 (-—) 40.6 (=) (+) 260 (=). 35% (42) S sett (+) 17.0 (—) 325 (+) 284 Before entering into a discussion of these results we shall consider the proportion of the phosphoric acid absorbed by the plants from the unre- covered phosphoric acid for the second year. On the Action of Various Insoluble Phosphates upon Rice Plants. 239 PO. With Ca O With Ca CO, Band eaves Unre- | P,O, in| Consumed from Unre- PO; in| Consumed from ef Ist year coverd | the total} unrecoverd P,O..|| coverd | the total junrecovered jg ae per ha,| '20s | ‘TOP % ofthe|| P,O; | crop | 13% of the phosphates, Bee vunre- PS 222 unre- kilo- | frame, | frame, coverd || frame, | frame, coverd grams, | grams. | grams, | grams. | P,O, |] grams.| grams. | grams. P2072. No PO, fo) 1.325 1.211 ae 25 1.507 1.641 erric pe 50 3.294 1.926 phosphate, 100 6.798 2.003 Ferrous 50 3.488 1.734 phosphate. 100 7.276 1.791 25 1.731 1.558 | 25 1.679 1.736 ‘ Aluminium 50 3-613 | 1.607 phosphate. 100 7.386 25 1.622 Calcium 59 3-545 phosphate, The figures of the two above tables decidedly prove, that in all the frames which received caustic lime in the first year, there were, with but few exceptions, apparently larger yields and also larger amounts of the residual phosphoric acid absorbed, than in the cases of the unlimed frames, but when a reference is made to the unfavorable results of the limed frames in the first year, this increase of the yield, was, in general, far from being sufficient to recover the diminution caused by the caustic lime in the first year. These results, therefore, prove that the action of the caustic lime has, by no means, favorably effected the assimilability of the phosphates. As to the frames supplied with calcium carbonate, it is seen that a slight increase of the crop was observed but in few frames, while in the majority, on the one side, a decrease and also a diminution of phosphoric acid consumption took place. 240 M. Nagaoka. As we have already seen from the results of the first year’s trials, the retarding action of the calcium carbonate was not so extensive as that of the caustic lime, but here, for the second year, we notice the assimilation of the residual phosphoric acid was interfered with considerably by further effects of the calcium carbonate left also from the preceeding year. With a view of obtaining still further data regarding the behavior of the residual phosphates to lime the series of the present investigation were continued for two years more. Leaving the details of the yield per frame for the appendix, the average of the equally treated frames are, for the third and fourth year given in the following two tables :— On the Action of Various Insoluble Phosphates upon Rice Plants. 241 THE YIELDS OF THE ‘THIRD YEAR. No of i, in the | second frame ee Kind of manures. eae rene Full Empty Total oe kilo- | frame Pee ere, | FOP landi Cat ), grams, grams. grams, | grams, | grams. grams. | grams, 1, 40, 79. | No P,O, without lime ra) 278.0 | 242.0| 2.8 | 522.8 2, 41, 80. | NoP,O, with CaO Oo 321.3 | 271.3| 4.0 | 596.6 738 Byeades tole i. ss as a Ee CaEOr fe) 276.7 | 228.2 2.5 507.4 (—) 15.4 7, 46, 85. | Ferric phos, with Ca O 25 1.191 | 399.3 | 340.1 ao | 743-3 220.5 "8, 47, 86. ” % » * 50 2.693 | 398.3 | 353-3 3-9 755°5 232.7 87s oy i ee ae too | r.o10 | 416.7 | 361.3] 5-3 | 783-3 260.5 10, 49, 88.) ,, , > CaCO, | 25 | 1.353 | 293-3| 2403] 3.3 | 5369 at PIs) 50; 89: 35 a - - 50 2.203 | 295.7 | 255.2 3.1 554-0 31.2 (eg AR is oh) i i ae 100 | 6.065 | 394.2 | 347.0] 5.6 | 746.8 224.0 16, 55, 94. | Ferrous phos, with{Ca O 25 1.268 | 326.7| 290-7| 4.0 | 621.4 98.6 He 56: 95. - aL Pee 5O |) 3.079 | 358.3| 311.8 | 4.1 674.2 | 151.4 18, 57, 96. » 33 A 5 100 | 6.800 | 408.7] 359.5 | 4.3 772-5 249-7 19, 58, 97-| » ec COn! 25) We T.coq | 264.1) |' 243.0 | . 3. | 5302 7-4 20, 59, 98. ” hs ae 50 3.283 | 339.3 | 296.2 | 3.7 639.2 116.4 at, 60;. ‘99. as a ES 100 | 6.053 | 343.4] 3087] 4-2 | 656.3 133-5 LS... | | ; 25, 64, 103. | Aluminium phos. withCaQ} 25 1.498 | 340.8 | 303-2 3.5 647.5 | 124.7 26, 65, 104. | % ou tee + 50 3.331 | 327-8] 286.0] 5.2 | 619.0 | 96.2 27, 66, 105. on Sm ot 3 100 6.778 | 388.5 | 350.2 Le 743.8 221.0 28, 67. 106. : 3 oy Ca Coy 3) 25 1.682 | 301.7 | 256.5 355 561.7 | 38.9 Wh fry) elle ugo 4) sg23'| 3063 | 2702) 3.7 | Soa} $74 39, 69, 108. es Sey he 3 1oo | 6.645 | 331.3 | 260.8 | 3.8 | 595-9 | 73-1 34; 73) 112: a phos, with Ca O | 25 1.554 | 352.6 | 287.0} 3.6 | 643.2 120.4 35, 74, 113 | ” » 9» 9 50 | 3.469 | 378.0 | 282.7 3.1 663.8 | 141.0 36, 75, 114. | + a3 ~ aS 100 7.047 415.9 | 375-9 4.2 | 796.0 273-2 aye 70, 125. » x , Ca co, 25 1.454 304-3 | 227-5) 4.1 535-9 13.1 Bow 775. 1x6, 5 Ad ; in | 50 3-175 | 246.7 | 307.2 3.3 557-2 34.4 39, 78, a | 100 | 6.396 | 330.5 | 281.0 | 3.9 | 605-4 $2.6 No of frames, M. Nagaoka. THE YIELDS OF THE FOURTH YEAR. _ PO; Average yield per frame. Surplus _ applied over the in the | frame Kind of manures. : St eae Bh Full Empty | Total without ilo. 2 | | grains grain, crop, and CaO grams. | grams. | grams, | grams, | grams, | grams. 40, 79. | No P,O, without lime. fe) 188,3 167.8 2.5 358.6 41, 8o. | No P,O, with Ca O Gon ip 23053 161.3 3.6 401.2 42.6 i a ne ase. fe) 196.0 148.7 2.6 3473 (—) 87 oo Jee a) Se 46, 85 | Ferric phos. with Ca O 25 286.0 226.0 3.0 515.0 156.4 47, 86. PR) Re 50 282.3 | 225.0 3.4 504.6 146.0 48, 87 | ee | Beet os 100 300.0 | 250.3 3.6 553-9 195.3 AQHRES.: || bss Linnean pees 25 267.0 | 207.3 3.1 477-4 118.8 50, 89. sy) ape me Ene 50 296.0 | 241.5 3.5 541.0 182.4 51, 90. » 3 , 3 100 308.0 | 260.3 3.8 572.1 213.5 Gia 13)! Ferrous phos. with Ca O 25 286.7 244.3 3-4 534-4 175.8 56, 95 ” ” ” » 50 315.3 260.0 3-7 579.9 220.4 57, 96. ” » » > 100 300.3 252.0 3.3 555.6 197.0 | 58, 97. | 3) fares ee COns por 260.7 | 206.3 2. 469.9 111.3 59, 98 ” , ” , 50 = |S(294.7 247.0 ah 544.8 186.2 60, 99. ay) A em ess | yelp iGo 281.0 | 224.2 3.4 508.6 150.0 64, 103. | Aluminium phos. withCaO, 25 289.3 243.8 33 536.4 177.8 65, 104. ” ” ’ ’ 50 300.0 242.2 3.6 545-8 187.2 66, 105 | ” ” ” » | 1co 327 3 282.5 3.6 613.4 254.8 67, 106 93 » wCaCO,! 2¢ 246.0 | 186.7 3.8 436.0 77-4 68, 107 ’ ssh Bs » 50 243-3 189.7 a2 436.2 77.6 69, 108 ” 53: a , 100 332.7 274.0 3.9 610.6 252.0 73% Vig Calcium phos, with Ca O 25 2098.0 238.0 2:3 539-3 180.7 74, 113 ‘9 9 » ” 50 Rhy fe) 274.7 3-7 602.7 244.1 75, 114 %» io nike 100 338.0 | 289.3 3.8 631-1 272.5 76, 115 s " » CaCO,| 25 | 3127 253.7 3.6 570.0 211.4 8, 77, 116 9 ee 59 320.7 | 253.0 3.5 577-2 218.6 39, 78, 117, | io Levee 100 | 315.7 | 259.7 3.5 578.9 220.3 | SL c . On the Action of Various Tusoluble Phosphates upon Rice Plants. 243 The figures contained in these two tables obviously demonstrate, in spite of the presence of the residual lime, still some favorable effects of the unrecovered phosphoric acid in the third and even up to the fourth year, but in the course of time, the variation of the yields between the three dif- ferent doses of each phosphate, became gradually narrower, although the frames which were manured more with the phosphates in the first year left also more of the phosphates for the successive crops. As to the after-effects of the lime it is seen from the frames without phosphoric acid that the caustic lime caused in both (3rd and 4th) years a gradation of the yield to a noted degree, while the calcium carbonate, on the other hand, caused some diminution of the crop when a comparison is made to that of the unlimed frame. The same fact is, in most cases, also observed in other frames which received the various phosphates and also two forms of the lime. This fact tends to prove that, although the calcium carbonate had a much weaker retarding action upon the effects of the phos- phates in the first year than the caustic lime, a gradual and unfavorable in- fluence of the former lasted even to the 4th year. This opposit action which exists between the caustic lime and calcium carbonate may be due to the natural character of both compounds. Thus caustic lime being endowed with stronger alkalinity than calcium carbonate, neutralizes more effectively the acid humus which is one of the most important factors in the solution of insoluble phosphates. The Carbonic acid which also plays an important role in the solution of the phosphates is further absorbed by the caustic lime. Caustic lime, when freshly applied, certainly neutralizes also the acid juice of roots decreasing thus also the dissolving actions of the roots. These are the main reasons for the injurious effects of caustic lime in the first year. On the other hand, the calcium carbonate had not such highly injurious action in the first year as the caustic lime had, but its actions were slower in the beginning and lasted longer. The calcium carbonate further is less soluble in the irrigating water than caustic lime, hence more is left for the following years. 244 M. Nagaoka. Before a general review of my results will be given, recent investiga- tions by /. Sutherst' wiil be mentioned. This author has studied the action of caustic lime and calcium car- bonate upon ferrous, ferric and aluminium phosphates and observed the following results. By the action of caustic lime, in presence of sufficient water upon ferrous, ferric and aluminium phosphates respectively for 1, 2 and 3 days, the solu- bility, in weak citric acid, of the phosphoric acid in these phosphates had extraordinarily increased in proportion of the lime applied; he has calculated the following percentage of the dissolved phosphoric acid. Hours, Ferrous phosphate. Ferric phosphate. Aluminium phosphate. 24. 75-42% 94-45. 64.33% 48. 85.45 5 96.38 ,, 69.31 5, Vie 85.88 ,, 96.55 5» 72.00 ,, In the same manner he has also tested the action of calcium carbonate, but the results were totally negative. Hence he infers: ‘“ From the above results it will be seen that it is essential that the lime applied in practice should be in the form of hydrate, the carbonate being of no value whatever.” The observations of Sutherst relate to trials in flasks, but while these were confirmed by myself, it is different with soils. K. Kawashima, formerly one of my students has carried out a trial under my direction on the series of the soils of my experimental frames which had received the various phosphates at the rate of 50 kilograms per hectare, that is the medium dose that we had applied. On the 12th of October, 1898, he took samples of the soil from the re- spective destinated frames of my first years trials above mentioned ina careful manner. He determined in each sample the phosphoric acid soluble in neutral ammonium citrate and also ina 5% acetic acid and obtained the following figures calculated in % of the dry samples. 1 Chemical News, Vol. LXXXV, No, 2210. p. 157. On the Action of Various Insoluble Phosphates upon Rice Plants. 245 No of frames 9% of the dry samples. from which the a8 ¢ : Kind of manures. ‘ ; : : samples were Phosphoric acid Phosphoric acid aen soluble in neutral soluble in the 5% of x ammonium citrate. acetic acid. 40. No phosphoric acid — 0,00224 41. As = » with CaO 0,04608 0,00214 42. 3 4 4 GROOR 0,05 162 0,cO291 44. Ferric phosphate 0,08575 0,00315 47. 3 7 with Ca O 0,05596 0,00199 50. ” ” » CaCO, 0,08377 0,00327 14. Ferrous phosphate 0,07315 0,0C491 iy a, nes ‘ with Ca O 0,04807 0,C0305 20. Hs y 5, CaCO, 0,06149 0,00402 23. Aluminium phosphate 0,07312 0,004 22 26. 75 with Ca O 0,05447 0,00270 29. 5 % 53) .Ca€O, 0,06977 0,00478 aa. Calcium phosphate 0,07604 0,00263 35. me with Ca O 0,06623 | 0,00289 38. 5s < 4) oa. CO. 0,07261 — Thus it is clearly seen that in no case a favorable action of the caustic lime upon the solubility of the insoluble phosphates had taken place, but on the contrary, the caustic lime, in all cases, diminished to a great extent the solubility of the phosphoric acid in both reagents, while the action of the carbonate was much weaker. This result sufficiently coincides with the actual yields in both, limed and unlimed frames and in the first year; hence it may be concluded that the caustic lime in soils has no beneficial action upon the insoluble phos- phates whatever, within a short time. ! 1 This fact was also proved in one of my recent trials (Cf, the previous article). 246 M. Nagaoka. In order to estimate the total effects of the liming the total sums of the four years’ crops obtained from the frames supplied with the various phos- phates and also with or without lime application, are given in the following table and further, the increase or decrease of the yields of the limed frames to those unlimed. | Retation of increase or decrease, ‘The respective yield of with- out lime = 1000. _ The total sum of the yield of four years. (grams.) | Phosphoric acid applied Kind of in the first | year. ae per ha, without with with with with kilograms, lime. CaO CaiCOs Ga'O Caco: No phosphoric acid. a) 2031.4 2122.3 | 1981.1 | (+) 44 (—) 25 25 2455.6 2760.0 2416.2 (+) 124 (-) 16 Ferric 50 2080.2 2914.8 2916.4 (—) 22 (-—) 29 phosphate. 100 3685.7 | 3403.5 3378.3. .|| (—) — 600 ae | 25 2379.5 | 2486.0 2417.6 (F)o- 4a ee II Ferrous 50 2795.9 | 2683.5 2739.8: I! (=)y goes tee phosphate. 100 3016.7 | 2974.6 3059.1 (—) 14 (+). 14 25 2439.6 2509.5 2384.8 (+) 29 (-) 23 Aluminium | 50 2882.4 2517.4 2492.9 (—) 127 (-—) 135 phosphate. | 100 3195.4 3032.2 3033.8 (=) 5. .] (hee | 25 2500.4 2482.0 2619.1 (—) 10 (+) 45 Calcium 50 2848.5 2696.6 2783.9 te 53 | (=) phosphate, ICO 3217.9 3089.1 3061.3 (—) 40 (-—) 49 From these figures it may be seen that the application of the caustic lime has caused but a slight increase of the rice crop in the frames without phosphoric acid and also in each of the smallest doses of the various phos- phates, excepting but that of the calcium phosphate. As to the action of the calcium carbonate, the results were quite irre- cular and no definite conclusion can be drawn here. | ) On the Action of Various Insoluble Phosphates upon Rice Plants. 247 In an aqueous mixture containing an excess of calcium hydrate the facts of Sutherst can be observed, whilst in soil the case is different. The caustic lime, before it can act upon the insoluble phosphate, can be absorbed mainly by the humus and partly be transformed into carbonate and silicate. Liming of the soils can therefore not be recommended for the culture of rice in presence of such conditions as in our paddy field. Finally I should recommend to the practical farmers an application of organic manures such as farm yard manures, acid cakes, peat &c., into over limed or freshly limed soils, as it would facilitate the absorption of insoluble phosphates by the plants and thus lead to a satisfactory harvest. —— —+ ~ep herent / et Note to the Preceding Paper. BY Oscar Loew. Soon after the conclusion of these highly valuable and laborious in- vestigations Prof. Nagaoka left for Europe and was not aware of the results Prof. Aso had obtained with rice grown in soil limed in various degrees and described in our Bul. VI, No. 2. Since the soil was quite of the same character and derived from the same locality, it may be permitted to add a few remarks. Aso has shown that for rice the best ratio of lime to ma- gnesia is =1 or nearly so, what agrees with the behavior of other cereals. Any increase either of lime or of magnesia beyond that proportion depressed the yield under otherwise equal conditions. Now the soil in question con- tains already lime and magnesia in nearly equal quantities, namely lime =0.9% magnesia =0.7%, hence the liming of that soil produced in Naga- oka’s case essentially the same results as in Aso’s trial. The effects of lim- ing, however, would differ on soils that contain a considerable surplus of magnesia over the lime content. 250 Appendix YIELDS PER FRAME, FIRST YEAR. Straw Full . Empty y No of grains grains No of frames. grams, grams. grams, frames. No P,O, and without lime. 46 85 1 267.0 243.0 5.0 8 40 330.0 266.0 67 47 79 426.0 359.0 10.5 86 No P,O, with Ca O. a a 48 2 262.0 205.0 6.0 87 41 246.0 174.0 4.7 8c 374.0 | 3030 | SHOR a) | | I No P,O, with Ca CO,. | ebore \ 3 260.0 212.0 | ia | 42 42 2650 196.0 | 50 8 1] 81 428.0 | 386.0 | 6.7 || nt | 50 Ferric phosphate. | 89 j 12 4 417.0 393.0 1277 P| 51 43 530.0 492.0 12.0 | i 82 517.0 469.0 13.5 | 9° 5 659.0 579.0 18.0 | 44 633.0 579.5 34.0 || 83 662.0 562.0 17.5 | 13 6 770.0 593.0 16.8 | 52 45 897.0 682.0 25 |i On $4 782.0 633.0 1217 | 14 i} sie ty | 53 Ferric phosphate with Ca O, | 92 7 | 344.0 281.5 9:3 | 15 Full grains Straw grams, grams, 420.0 351.0 405.0 370.0 382.0 469.0 581.0 608 0 523.0 Ferric phosphate with Ca COg,. 408.0 357-0 406.0 aaae 522.0 470.0 640.0 482.0 552.0 494-0 635.0 559.0 663.0 510.0 670.0 503.0 763.0 613.0 Ferrous phosphate, 429.0 412.0 494.0 405.0 506.0 409.0 411.0 400.0 6440 473-0 602.0 503.0 585.0 513.0 Empty grains grams, 14.2 9.0 9-7 25.7 14.0 24.0 24.5 22.0 16.8 10.7 12.5 15.2 9.7 13.0 18.0 16.0 No of Straw frames. grams, 54 616.0 93 696.0 ° Ferrous phosphate with Ca O, 16 253.0 55 309-0 94 428.0 17 310.0 56 383 0 95 414.0 18 383.0 57 497.0 96 534.0 Ferrous phosphate with Ca COg3. Appendix. YIELDS. PER FRAME, FIRST YEAR. Fall grains grams, 490.0 586.0 212.0 263.0 370.0 256.0 349.0 352.0 342.0 432.0 426.0 Empty grains grams. 27.5 13-7 4.5 7:7 9.0 6.7 7:5 9.0 6.7 10.0 10.0 19 2098.0 251.0 6.0 58 - 462.0 387.0 13.2 97 478.0 409.0 10.0 20 520.0 401.0 127 59 522.0 434.0 14.7 98 575.0 500.0 12.5 21 609.0 541.0 13.7 60 619.0 555.0 17.3 99 643.0 522.0 16.0 Aluminium phosphate. 22 374.0 312.0 9.0 61 449.0 329.0 12.5 25 (197.0) 64 333.0 103 542.0 26 264.0 65 364.0 104 552.0 27 430.0 66 407.0 105 525.0 Aluminium phosphate 28 318.0 67 406.0 106 506.0 29 361.0 68 447.0 107 572.0 30 618.0 69 506.0 251 Meneame} ) eeeeast | tigen frames, grams, grams. grams. 100 548.0 14-5 23 521.0 14.3 62 597.0 16.8 IOI 592.0 ners, 24 647.0 16.7 63 567.0 16.0 102 735.0 foe (133.0) (4.2 286.0 7-3 440.0 14.7 191.0 4.0 306.0 7-2 468.0 11.2 403.0 11.7 340.0 8.0 467.0 11.5 with Ca CQ,. 267.0 5.8 344.5 7-2 440.0 $8.7 312.0 g.2 403.0 6.5 518.0 13.5 553-0 14.0 448.0 15.0 252 Appendix. YIELDS PER FRAME, FIRST YEAR, a Straw Full Empty b Straw Full Empty No of grains grains No of grains grains frames, grams, grams, grams. frames, grams, grams, grams, 108 610.0 543.0 12.7 35 302.0 232.0 6.5 74 372.0 283.0 13-7 Calcium phosphate. , 113 666.0 538.0 21.0 31 387.0 345.0 7.0 36 445.0 367.0 9.8 70 546.0 475.0 14.0 75 500.0 428.0 D7 109 567.0 520.0 12.0 114 585.0 468.0 16.0 32 655.0 555.0 17.7 Calcium phosphate with Ca CO,. 71 521.0 425.0 19.0 meas 110 679.0 507.0 19.7 37 437.0 390.0 11.5 33 755.0 653.0 25.2 76 452.0 408.0 Be 72 663.0 510.0 24.5 115 638.0 492.0 19.2 Ill 728.0 603.0 18.7 38 547-0 481.0 T5.2 Calcium phosphate with Ca O, 34 340.0 280.0 73 3540 258.0 112 531.0 398.0 Appendix. 253 YIELDS PER FRAME, SECOND YEAR. A LT | Straw Full Empty | a Straw Full Empty No of grains erin | Noof grains grains frames. grams, grams. grams, frames. grams, grams, grams, No P,O, no lime. 46 391.3 293.0 5-7 85 423-7 319-5 7-5 I 250.5 216.0 7.0 8 383.3 320.0 5.0 40 324.7 226.0 5:5 47 423.3 325.0 6.5 79 304-3 199.0 3-7 86 465.5 361.5 10.9 No P,O,!wlth Ca O. 9 471.3 368.0 7.7 48 408.7 496.0 7.0 2 258.5 191.0 4.0 87 512.8 418.0 10.2 41 363.2 284.0 4.5 80 406.3 270.0 9.2 No P,O, with Ca CO,. 3 250-7 182.0 4.2 42 Si 7 218.0 4.7 SI 356.3 277.0 8.5 Ferric phosphate. 4 248.8 181.0 3-4 43 347.8 265.5 4.8 82 409.8 305.5 9.0 5 324.8 262.5 5.0 Ferrous phosphate. 44 344-5 263.0 4.5 83 396.7 302.0 9.0 13 252.8 187.0 3.5 6 396.7 346.0 6.2 52 326.3 236.0 I 45 463-3 385.5 75 gl 351-3 269.0 6.5 $4 499.3 407.5 7.4 14 265.7 204.0 4.1 ; ae | 53 363.8 261.0 6.0 Ferric phosphate with Ca O. | 92 367.8 291.0 6.5 7 361.3 281.5 | 5.7 | 15 353.8 297.5 4.9 ll 254 Appendix. YIELDS PER FRAME, SECOND YEAR, No of sat ae ae frames, grams grams. grams. 100 342.3 240.0 8.0 23 340.3 240.0 4-7 62 403.0 312.0 8.2 101 345.0 263.0 5.8 24 374.8 317.0 5.2 63 406.0 299.0 9.0 102 370.5 303-3 6.1 Aluminium phosphate with Ca O. A Straw Full Empty No of grains grains frames, grams, grams, grams, 54 454.5 347.0 7-9 93 431.8 353-0 6.5 Ferrous phosphate with Ca O. 16 341.8 260.0 6.5 55 434-7 334.0 9.6 94 412.7 328.0 6.9 17 342.8 261.5 5-7 56 440.8 321.5 8.2 95 457-8 356.0 9.4 18 340.6 267.0 57 57 429.0 324.0 9.4 96 454.0 362.0 6.9 Ferrous phosphate with Ca CO,. 19 222.8 154.7 Si 58 346.0 256.2 5-5 99 378.0 265.2 6.5 20 249.5 188.0 3.4 59 337-5 253.0 6.2 98 360.8 271.0 6.0 21 377.3 311.0 5:7 60 427.0 322.0 2 99 417.0 320.0 7-0 Aluminium phosphate, 22 262.5 177.0 4.0 61 356.7 167.5 6.5 25 320.3 249.0 4.9 64. 434-3 307.0 10,0 103 381.8 306.5 6.2 26 342.8 269.0 5.2 65 419.8 319.0 8.2 104 B3a°2 285.0 8.1 27 460.3 356.0 6.5 66 453-5 342.5 8.4 105 426.3 359.0 95 , Aluminium phosphate with Ca COg. 28 260.7 186.0 3.5 67 303.8 219.0 7h} 106 320.7 240.0 8.0 20 278.8 204.0 4-4 68 368.0 276.0 6.9 107 366-0 275.0 _ 8.2 30 379.5 309:0 5-1 69 440.8 334.0 99 Appendix. 255 YIELDS PER FRAME, SECOND YEAR. Straw Full Empty 2 Straw Full Empty No of grains grains No of grains grains frames. grams, grams, grams, frames. grams, grams, grams. 108 392.8 287.0 9.5 35 339-3 245.0 4.7 74 364.5 240.0 7.2 Calcium phosphate. 113 368.5 279.0 7.9 31 242.8 174.0 BG) 36 Stile 284.0 6.5 70 356.8 262.0 7.4 75 444.3 333-0 75 109 348.3 250.0 7.9 114 394.0 300.0 8.7 32 307.8 235-5 4.7 Calcium phosphate with Ca CO,. 71 369.8 278.0 8.2 110 361.8 253.0 10.0 37 a3 322.7 266.0 4.5 76 72 396.0 312.0 6.0 115 III 325.0 246.0 6.9 38 Calcium phosphate with Ca O. 256 Appendix. YIELDS PER FRAME, THIRD YEAR. EE ES Bay Straw Full Empty | Straw Full Empty No of grains grains No of grains grains frames. grams, grams, grams. frames, grams, grams, grams, No P,O, no lime, 46 423.0 349.5 3.9 85 438.0 379.0 3-9 I 211.0 164.0 2.5 8 306.0 279.0 3.4. 40 3260 294.0 2.9 47 439.0 382.0 4.0 79 297.0 268.0 2.9 86 450.0 399-0 4.2 No P,O, with Ca O. 9 352.0 321.0 4.5 48 470.0 377-0 5-9 2 252.0 207.0 55 87 428.0 386.0 5:4 41 353.0 295.0 2.9 a 359.0 ba 35 Ferric phosphate with Ca CO,. No Jer with Ca CO. | Io 217.0 160.0 3.0 3 216.0 185.0 2.4 49 sane see 3-4 42 348.0 275.0 2) | ae 337-0 Ee 3-4 81 GG 224.5 oe Il 187.0 122.0 212 50 306.0 303.5 3.0 Ferric phosphate. 89 394.0 340.0 4.2 : 6. 4 252.0 222.0 oH) si 364-0 a ‘ I 320 15.0 f 43 273.0 244.0 4.2 5 aoe 315 57 $2 318.0 264.5 2.2 a 497-9 ae +2 5 247.0 211.7 3.5 ; Ferrous phosphate. 44 387.0 306.0 3.5 83 386.0 345.5 3.5 13 248.0 221.0 3.5 6 347.0 338.0 3.9 52 247.0 206.0 4:5 45 398.0 355.0 Biz ol 365.0 284.0 4.0 &4 426.7 387.0 Biz 14 279.0 243.0 3.4 53 295-5 233.5 3:2 Ferric phosphate with Ca,O, =—- 92 362.5 304.0 2.9 7 337.0 291.7 | 3:7 15 265.0 194.0 3-7 t No of Straw frames. grams, 54 309-7 93 351.0 22 Appendix, YIELDS PER FRAME, THIRD YEAR, Full grains grams. 208.7 310.7 Ferrous phosphate with Ca O, 238.0 347.0 395-9 243.0 374.0 458.0 407.0 368.0 451.0 201.0 330°0 341.0 212.0 344.0 379-5 361.5 350.0 367.0 Ferrous phosphate with Ca 261.5 278.5 312.5 317.0 328.0 373.0 315.7 325.0 359.5 Aluminium ae 334-0 302.5 238.0 220.5 261.5 284.7 296.0 308.0 269.7 315-5 341.0 281.0 248.5 Empty grains grams, 3-9 3:5 CO,. No of frames. 100 Straw grams. 384.7 347-9 380.0 346.0 316.0 3838.0 413.0 i) N Full Empty grains grains grams, grams, 304.7 4.5 286.0 4.2 325.0 4.2 292.0 25 240.0 7 343°5 4-4 358.0 4.2 Aluminium phosphate with Ca O. 273.0 212.0 2 373-0 354-5 3-5 376.5 343.0 3-7 27-5 193-5 3-2 388.0 358.5 27 368.0 306.0 8.7 364.5 340.0 5-9 48.0 320.5 5.0 453-0 390.0 4.5 Aluminium phosphate with Ca CO 309.5 262.5 3.7 306.0 266.5 3.4 289.5 240.5 3-5 265.0 222.0 4.4 283.0 251.5 3.2 371.0 337-9 3-4 327.0 193.0 4.0 325.0 283.0 ey 258 Appendix. YIELDS PER FRAME, THIRD YEAR. i é Straw Full Empty : Straw Full Empty No of grains grains No of grains grains frames. grams. grams. grams, frames. grams. grams, grams. 108 342.0 306.5 Buy 35 400.0 223.0 45 74 393-5 355-5 35 Calcium phosphate. ‘ 113 307-5 299-5 4-4 31 271.0 225.0 2: 36 444.0 383.0 4.7 , 70 2581.0 215.0 2.5 75 366.7 359.7 4.2 | 109 314.5 255-5 Bis 114 437.0 385.0 avy | 32 306.0 241.0 2 Calcium phosphate with Ca CO,. . 71 324.0 260.0 2, 110 313.5 244.0 4.4 37 2.9 33 395.5 328.5 5.4 76 4.5 ’ 72 364.0 291.5 3-9 115 5.0 . 111 367.0 272.0 3.0 38 3.7 i Calcium phosphate with Ca O. 34 369.5 311.0 73 365.5 300.5 112 323.0 249.5 ————— Appendix. YIELDS PER FRAME, FOURTH YEAR. Straw Full Empty No of grains grains frames. grams, grams, grams, No P,O, no lime. I 206.0 207.6 2.0 40 137.0 95-5 2.1 79 222.0 200.2 eGee | No P,O, with Ca O. 2 272.0 200.0 35 41 225.0 159.0 3.0 80 212.0 155.0 4.0 No P.O. with.Ca.CO,. 3 183.0 143.0 118y 42 140.0 87.0 2.0 Sr 268.0 216.0 4.1 || Ferric phosphate. 4 215.0 169.0 17, 43 146.0 98.0 2.2 | 82 281.0 236.0 3.2 ls 225.0 176.0 2.4 | 44 235.0 175.7 2.1 83 278.0 243.0 4.0 337-0 298.0 2.9 II 45 372.0 291.0 35 | 84 300.0 262.0 48 | | Ferric phosphate with Ca O, | i 7 2098.0 238.0 2 | No of Straw frames, grams, 46 290.0 85 282.0 8 264.0 47 289.0 86 204.0 9 294.0 48 299.0 87 307.0 Ferric phosphate with Ca CQ,. fe) 306.0 49 218.0 88 277.0 II 257.0 50 399.0 89 332.0 12 286.0 5t 356.0 go 282.0 Full grains grams. 216.0 224.0 210.0 240.0 164.0 218.0 205.0 319.0 200.0 Ferrous phosphate. re 158.0 52 262.0 QI 267.0 14 241.0 53 254.0 92 210-0 15 282.0 94.0 211.0 Empty grains grams, _—-..1.3. SS | | | 260 61 Straw grams, Ferrous phosphate with Ca O. 294.0 317.0 249.0 318.0 301.0 327.0 260.0 355-0 286.0 Ferrous phosphate with Ca CO Zio 272.0 233.0 296.0 268.0 320.0 220.0 293.0 330.0 Aluminium phosphate, 274.0 259.0 Full grains grams, 243-5 258.6 231.0 266.0 225.0 289.0 214.0 303.0 239.0 255.6 220.0 Appendix, Empty grains grams, 3° tN Se iS) © YIELDS PER FRAME, FOURTH YEAR. nae Straw frames. grams, 100 217.0 23 274.0 62 282.0 IOI 299.0 24 3150 63 330-8 102 290.0 Full grains Empty grains grams, Aluminium phosphate with Ca O, 25 323.0 64 350.0 103 295.0 26 312.0 65 313.0 104 275.0 27 327.0 66 357.0 105 298.0 214.5 260.0 257.0 253-7 262.0 211.0 265.5 301.0 281.0 Aluminium phosphate with Ca CO,. 28 281.0 67 250.0 106 207.0 29 206.0 68 224.0 107 300.0 30 311.0 69 361.0 231.0 177.0 Appendix. 261 YIELDS PER FRAME, FOURTH YEAR. eh cee | Note | geting | CmEe frames. grams. grams. grams. frames. grams. grams, grams, 108 326.0 251.0 4.5 35 271.0 211.0 3-0 a 74 355.0 395.0 4.0 Calcium phosphate. 113 347.0 308.0 3-7 31 276.0 219.0 PAG | 36 274.0 217.0 3.4 70 220.0 178.0 ZG 75 388.0 335.0 4.4 109 299.0 218.0 3:9 114 352.0 316.0 3. 32 301.0 242.0 2.1 He es ce a Calcium phosphate with Ca CO,, 110 215.0 160.0 Se! 37 254.0 194.0 aS | 33 320.0 273.0 20 76 317.0 240.0 4.5 72 359.0 303.0 4.7 115 367.0 327.0 4.0 | Iif 270.0 221.0 a2 38 323.0 261.0 2.4 | is 353.0 261.0 4.5 Calcium phosphate with Ca O, —————————— OO , _—- HO & —- - On the Effects of Soil Ignition upon the Availability of Phosphoric acid for Rice Culture in Paddy Fields. BY M. Nagaoka. My earlier experiments carricd out in conjunction with my colleagues, proved the fact that in the paddy field of our college the application of soluble phosphoric acid in the form of sodium phosphate and superphosphate often gave a rather inferior yield than precipitated calcium phosphate and further it was observed that the action of those soluble phosphates soon faded away, although the residual phosphoric acid, proved from the analytical data, to be present in such a quantity that it could produce still a full crop of rice. At first we supposed that this might be due to the transformation of the soluble phosphoric acid into iron and aluminium phosphates. However, my later trials on the manurial action of the various phospha- tes, such as ferric, ferrous, aluminium phosphate & upon rice plants proved, contrary to my anticipation, that, in the first season, such insoluble phos- phates displayed a most satisfactory good action upon the plants, the yield of the ferric phosphhate being even larger, and also these of ferrous and aluminium phosphate being less only 24 %% and 27 9% respectively than that of the double superphosphate, and, in the second year, their residual phosphoric acids gave still comparatively better results as also compared to that of the double superphosphate. These facts sufficiently show that our former assumption was ‘not quite correct. There must exist another cir- cumstance that renders the phosphoric acid insoluble and unavailable. I suspected that the rich humus content (119%) of our soil has something to do with that phenomenon. As to the original phosphoric acid of our paddy soil, we have often found it to be as much as 0.49% in the dry samples, which quantity exceeds 204 M. Nagaoka. that present generally in Japanese paddy fields, and would according to our calculations, be quite sufficient to produce a medium rice crop every year for more than two hundred years without any new supply of phosphoric acid. Nevertheless our practical investigations have always shown results entirely contradictory, since manuring with nitrogen and potash alone— leaving out a fresh supply of phosphoric acid to this soil, gave, as a rule, a very unsatisfactory crop—resembling that obtained on entirely unmanured soil. The supposition above expressed led me to investigate the behavior of ignited paddy soil (1899). It was important to determine how much phos- phoric acid might be rendered available by the process of ignition-or incineration of its humus content. I determined in both original and burnt soil the phosphoric acid soluble in water, in neutral ammonium eitrate, in 5 % acetic acid, in 1 9§ citric acid, in 1 9% oxalic acid and in saturated carbonic acid solution, and further the total phosphoric acid extracted by a hot hydrochoric acid of 1,15 sp. gr. The sample of the soil, which served for these trials was taken from a part of our experimental paddy field. The well dried soil was sifted through a sicve with meshes of 1/2 milimeter. The burnt soil was prepared from a part of the same sample, by incineration for fifteen minutes to faint redness, by which process the loss of ignition was 21,123 % of which 10,92 % was that of the genuine humus. In each trial, I took 50 grams of the original soil and 39.439 grams from the burnt soil, which quantities corresponded to 50 grams of the original sample. The digestion of the weighed soils lasted for seventy two hours stirring from time to time and in the respective filtrate, the phosphoric acid was determined in the usual way. The results will be seen in the following table :— ————————— Ll oe Effects of Soil Ignition upon the Availability of Phosphoric acid. 26 ut Dissolved phosphoric acid in 9% of the dry soil. The rate of Kind of solvents. ’ : Original soil. Burnt soil. ete Ilydrochloric acid (sp. gr. 1.15)...... 0.475 0.499 Got MPSA CE oes vy sd dans canncvigernn' trace trace —— AARON “CHEATS, 2, ...0.ccneseesness 0.046 0.076 63-9 JHE NESEY) CIR (CR) Re ee 0.145 0.167 15.2 Pine AEM (Eg) scinnsy (99 33 ” 39 ” 5; 22, 39. 2 33 bed ” ” 6 ” ” ” ” hd ” 3 ” ” ” ”” On the 25th, after a sufficient application of distilled water so as to to give the soils a state of mud, the general manures in the form of sulphates, were added and the soils then agitated thoroughly. On the 30th, the young rice plants (Variety Satsuma) were transplanted in such a manner that each pot received 3 bundles, each consisting of three healthy in dividuals of equal size. After the transplantion, the same level of water was procured for all pots, a supply of some distilled water from time to time being procured, but after the blossom of the plants the application of water was diminished, so that the soils maintained only sufficient humidity for the ripening of the seeds. On the 15th of November the rice plants were harvested when they had attained the state of milk ripeness, and left to become air-dry. The yields per pot and also the averages of the three parallel pots were as follows :— 268 M. Nagaoka. No. No. Shea Grains. Average per pot, of Kind of soils. of pots, panicles, eae er, Straw, Grains, | Total crop. er. er. er, I 12 17.0 10.5 Original soil with no . 18 14 162 iy) 16.17 10.63 26.80 manures, 35 15 15.3 9.7 2 36 38.2 Baur Original soil with 19 29 Bat 19.9 39.07 21.70 60.77 manures. 36 30 38.9 21.5 2 13 Es 3.0 8 Kg. burnt soil with 20 12 6.2 12 9.27 2.13 11.40 manures, 37 12 10.3 Die 4 ’ ye 33 45.8 26.7 4 Kg. burnt soil with 21 34 42.3 24.7 41.30 23.47 6477 manures. 38 25 35.8 5 : 54 68.3 2 Kg, burnt soil with 22 45 64.8 111.83 manures, The moderate doses of the burnt soil had therefore a considerable action upon the yield of the rice plants. As all the pots, excepting those of with- out manures, had received nitrogen and potash in sufficient quantities and as all were treated in a similar manner, the above differences in the yield, must be due, according to the law of minimum, to the influence of the phosphoric acid in the soils. The effect of the phosphoric acid in the ignited soils will be noticed more conveniently in the following calculation, in which we assume the plus yield of the original soil with manures over the yield of that not manured, as 100. Effects of Soil Ignition upon the Availability of Phosphoric acid. 269 Plus yields, | Ration of increase. = att | Total crop. Grains, >: eee aa in Yotal crop. | Grains, Original soil with manures, |(+) 33.97 (4), “18.07 | 100 100 rs) USI [SLEVIN E Nea aeApe pre Ber (—) 15.40 (—) 8.50 | (—) 45-2 | (—) 76.8 Asa ss 53 Peo cetochaes ask (ae) Bay (4) 12.84 | (+) 111.8 (4) 116.0 2... “py. Ie eats (+) 85.03 (2)) Shas | | It will be noticed from these figures, that the smallest dose of the ignited soil gave more than the double plus yield over that of the original soil even with the same manures. By increasing the amount of burnt soil to 4 Kilograms per pot the yield decreased, while this decrease was still greater when burnt soil alone served for the crop. I shall further consider here how much phosphoric acid was extracted by the crop from each pot; the results of the chemical analysis of the crops gave the following figures. Phosphoric acid in Surplus phosphoric | ‘The surplus of the Kind of soils. the total crop. acid. | original soil with gr. er. | manures = 100. Original soil without manure. 0.0682 — —- = = with manure, 0.1053 (+) 0.0371 100 S Berburnt’soil, os 2..2.0-.... 0.0341 (-) 0.0341 r(—) 91.9 : Ate es soe ast AES ete ME 0.1184 (+) 0.0502 (+) 135.3 | Das, t3 “He, » Seno GReReEee ee 0.1930 (+) o1258 | (+) 339.1 Thus it will be attested that the numbers of the last calumn almost agree with the increase in grains, and this also proves that from the moderate quantity of the burnt soil admixed to the original soil more phosphoric acid is rendered available to the rice plants, and in consequence of which the increase of grams attains almost the same proportion of the phosphoric acid absorbed. But when the quantity of the ignited soil increased to a certain extent over that of the original soil, the availability of the phosphoric was much decreased, beside, with the ignited soil only the phosphoric acid absorption and the yield were both uncomparably 270 M. Nagaoka. diminished. Hence we may suggest that the economy of the phosphoric acid in burnt soils requires some precautions with special regard to the proportion of a burnt soil to the unburnt, the character of which and contents requiring careful consideration. It is highly probable that the acid humus contained in the original soil played an important role, inasmuch as it acted as a solvent of the phosphoric acid of the burnt soil; this point of view will be further explained later on. This series of the experiments was carried on for the second year in the hope of obtaining some further informations on the ignited soil. All pots, excepting these without manures, received, on June 21 (1901) the same quantity and the same form of nitrogen and potash as the preceeding year. On July 1., the young rice plants were also planted just in the same manner as before. The crops were harvested on the 26th of November with the following figures of the air dry matter. I added in the last column the numbers, resulting from chemical analysis, of the phosphoric acid contained in the total crop per pot. No. Straw. | Grains, Average per pot. pee ic acid of Kind of soil —Suaw. | Grains [Tos epee pots. gr. gr. gr. gr. gr. : gr. I | Original soil without manure. | 15.0 6.0 18 + - A: - 16.0 7.2 15.67 7.10 22:77 0.02782 35 » » 9 ” 16.0 8.1 2 | Original soil with manure. 15.0 9.8 19 uw eS fs 14.0 6.3 16.33 8.17 24.50 0.02837 36 » ”» 99 20.0 8.4 3 | 8 Kg. burnt soil with nanure,| 3.1 om 20 |] 5 » » » 9 ” 3.5 OI 4.03 0.27 43 0,00272 S7 Nish 293, oat SFE 3 5.6 0.6 4 | 4 Kg. burnt soil with manure. | 31.1 21.6 . 4 el Oe ek Ge i + 48.6 38.1 41.27 ara3 72.40 0,0816 38) ww agt as Pah OH ” 44.1 33-7 5 | 2 Kg. burnt soil with manure, | 24.0 16.7 22] 55 99 gh hss ” 33-5 16.7 30.17 15.97 46.14 0.07096 ci Tete MTP ee Pay, 9 33-0 14.5 Effects of Soil Ignition upon the Availability of Phosphoric acid. 271 It is seen, in general, from the above results that the yield, excepting but one (4 kilograms of the burnt soil) and the phosphoric acid consumption more or less actually diminished as compared to the former year’s, and this proves the fact that the stock of the phosphoric acid in all pots was already in the first year exhausted toa great extent, and for the coming year but little of the nutriment was left in an assimitable form. Yet when a close comparison is made it is well proved that the ignition of the soil had still a considerable influence upon the rice crop even in the second year, for, the yield of the four kilograms of the burnt soils attained almost to three fold and that of the two kilograms, two fold of the yield of the original soil with sufficient general manures. As to the pots which were supplied with 4 kilograms of the burnt soil, the phosphoric acid extraction by the plants was not so great as in the first year, but wee see, on the contrary, the actual yield was augumented as much as 12% more than that of the first season. This proves the fact that with the proportion of 4 kilograms of the burnt soil to the same quantity of the original one, the phosphoric acid contained in the former soil is rather slowly and less absorbed in the second year but this nutriment is utilized by ths rice plants for organic matter production in a most satisfactory manner. The pots, which received the burnt soil only, exhibited again a most miserable condition and gave quite an insignificant yield in the second year. This is due most probably to the insufficiency of some acid humus and further, to the poor mechanical condition caused by the ignition. From these trials with ignited soil we may now safely conclude that, in some soils rich in humus, there exists certainly more or less phosphoric acid in form of organic compounds which can be utilized in practice by ignition to a certain extent. In 1901, G. Datkuhara'! and 7. Hanaz have published some very 1 Report of the Agricultural Experiment Station, Tokio, Vol. 20. 272 M. Nagaoka. interesting and useful articles upon the so called “ Burnt soil manure.”’? The authors have carried out separately some experiment on the said manures which were specially prepared by themselves and they observed that in such gently roasted manures, the solubility of the phosphoric acid in several weak acids had considerably increased and the proportion of this increase was, in average, culculated as much as 37% over the soluble phosphoric acid of the original soils (that is the soils which served as the material of the smouldered manure). Besides, Datkuhara has even ob- served that soil poor in humus can be enriched with soluble phosphoric acid by gently roasting it. These. authors have further undertaken several ex- tensive comparative pot experiments with such manures upon rice as well as barley and obtained similar results which proves that the phosphoric acid in the burnt soil manures gave, in general, a much better havest than that without this manure. Dazkuhara concluded that the increase of the solubility of the phosphoric acid in a burnt soil manure is partly due to the destruction of some organic phosphatic matter and partly to the influence upon tne mineral phosphatic constituents of tne original soil. When we take into consideration the results obtained by the above two authors as compared to those obtained by myself, the part is still further confirmed that the existence of the phosphoric acid in form of some organic combinations in soils rich in humus is possible and ignition or as well as gently roasting (smouldering) of these soils can secure some profit. fl 6Sertes. Are caustic lime, calcium carbonate and potassium carbonate beneficial to the assimilation of the phosphoric acid in a burnt soil ? These series of the trials were performed in accordance with the prece- eding experiments with the view to determine whether caustic lime, calcium 2 The burnt soil manures are, according to CG, Daikuhara, generally prepared by a very slow smouldering or gently roasting of some soils such as muds from ditches, soils from mountains and plains arable soils from certain cultivated fields etc, and these manures are extensively and particularly used tor the cultivation of various agricultural crops in the south western provinces of Japan, Effects of Soil Ignition upon the Availability of Phosphoric acid. 273 carbonate and potassium carbonate have some useful action or not upon the efficacy of the phosphoric acid in a burnt soil. The treatment of the rice plants was the same as usual, but all the pots received either caustic lime or calcium) carbonate at the rate of 1000 kilograms of CaO per hectare. These lime compounds were applied in the first year only and they were mixed carefully into the weighed soils before the latters were filled into the pots. As to the pots of the potassium carbonate, we had not enough burnt soil to carry out the whole series, so I tried but for 4 kilograms of the ignited soil, the quantity of which being the medium ration of this series. These pots received, in the first year, besides the general manure 100 kilograms of K,O per hectare in the form of potassium carbonate, and in the second year, the same rate of the carbonate, but not the potassium sulphate as general manure. The crops for both, first and second year were likewise harvested the same days as that the first series. To avoid too much complication of the numbers, I shall simply give the averages of the parallel trials in the following table in which the actual quantities of the phosphoric acid absorbed by the rice plants from the soil per pot are also given. 274 M. Nagaoka. First year. Second year. = : Phosphori Phosphori Kind of soils, Average per pot, Ned en TEE Average per pot. ae d ee || (0) Fl (0POYS), |) fil) TID, Straw. |Grains.| Total Straw. |Grains.| Total gr. er, crop. er. gr, gr, crop. gr. With caustic lime. Original soil with manure. | 41-00 | 19.37 | 60.37 0.1026 15.10 7.60 | 22.70 | 0.01341 - , not 8 Kg. burnt soil. ............ 8.07 | 1.53 | 9-60] 0.0074 4.93 | 9.20] 5-13 | getermined As, 4 Bae aha one taee 38.00 | 21.60 | 59.60 0.0765 41.00 | 26.40 | 67.40 | 0-0725 Ph ee Pseawinss Sensis 47:30 | 34.45 | 81.75 0.1227 23.00 | 12.00 | 35.00] 0.0576 With calcium carbonate, Original soil with manure, | 41.40 | 17.70] 59.10 O.1015 10.50 5:15 | 15.65 | 0.01538 rodeo Jobst A EON I an Aamacdenc 8.40 220] 10.60 0.0108 2.80 fo) 2.80 gurers 4» ” eet Se 39.80 | 20.20 | 60.00 0.0831 47°50.| 28-95 | 76.45 | 0.1047 Ti Sin eae ai PO: kecrcepocaee 52.20 | 36.05 | 88.25 0.1364 30.25 | 17.15 | 47.40] 0.04828 With potassium | carbonate, Arise, DUPNb SOlece ss tence. 22.25 | 15.60 | 47.85 0.0516 30.50 | 25.4 55.90 | 0.05919 From these figures the part can be learned that, in spite of the presence of the alkalies, the proportions of the increase or decrease between the yields of the burnt and not burnt soils attained almost the same degree as in the first series, which fact proves also that the best action of the phosphoric acid in the soil is obtained by mixing burnt and unburnt soil in the proportion of 1: 3. But when these figures are compared to those obtained in the two years of the first series, we have again a most obvious evidence that by the application of some alkaline compounds, the yield in all soils, whether burnt or unburnt, was more or less diminished and the phosphorie acid consumption by the plants was also equally affected. In order to show this point still more clearly, I calculated in the following table the actual diminutions of yields as well as the diminutions of the phosphoric acid consumption in consequence of the alkali application. I‘urthermore with the aid of these numbers the percentage diminution in Effects of Soil Ignition upon the Availability of Phosphoric acid. 275 each pot was calculated when the total crops of the pots not supplied with any alkali were, for the first and second year, assumed respectively to be 100. | First year, Second year. ze add Actual diminu- | Percentage di- Actual diminution Percentage dimi- (Hag tion per pot. minution, per pot. nution, Yield. O2P 5. | viela. P,0,. Yield. HeOes Vield, P.O. gr. er. z gr. gr. 2) 5 With caustic lime. Original soil. 0:40 | 0.0027| 0.7 2.6 1,80 0.01496 73 ey) 8 Kg. burnt soil. 1:80 | 0.0267} 15.8 | 78.3 |I(+) 0.87 = ek 20-2 —= 4» % 9 5-17 | 0.0419 8.0 35-4 5.00 0.0091 6.9 1 plane 3 Pr 30.08 | 0.0703] 18.5 36.4 11.14 0.01336 24.2 18.8 With calcium carbonate, Original soil. 1.67 | 0.0038| 2.8 3.6 8.85 0.01298 36.1 45.8 8 Kg. burnt soil. 0.80 | 0.0233] 7.0 68.3 1.15 — 34.8 oe A, + Pp 4.77 | 0.0353) 7-4 | 29.8 |(+) 4.05 ((+)0.0231 (+) 1.6 |(+) 28.3 2s 5 " 23.58 | 0.0566] 21.1 29.3 |\(+) 1.26 0.02268](+) 2.7 31.8 With potassium carbonate. 4 Kg. burnt soil. 16.92 | 0.0568| 26.1 47.1 16.50 0.02244 22.8 27.5 As it is seen in the above figures, the total yields and phosphoric acid consumption in the first year were both affected by the alkali application to a certain extent and the percentage diminution of the phosphoric acid consumption was far more greater than that of the total yields. In the second year this decrease was still observed in same pots, while in others that had received calcium carbonate, there was a slight surplus over the yield of the control (not limed pot), but this surplus was not sufficient to compensate the loss in the preceding year. In general, some unfavourable action of the alkali still prevailed in the second year in the majority of the pots. As to the difference of the effects of the caustic lime and calcium carbonate, it is seen the former acted more 276 M. Nagaoka. powerfully than the latter in both years, while the potassium still more so. It is evidently the neutralization of the acid humus in our paddy soil as well as of the acid juice of the plant roots which diminishes the availability of the phosphoric acid. Since ignited soils have, in general, a rather strong alkaline character, the poor results obtained on burnt soil alone without an addition of unburnt, are also explained. Hence it may be recommended to add to burnt soil a sufficient amount of humus or organic matter yielding humus. Compounds of an alkaline nature are injurious to the rice grown in paddy soils. ——————_——— BUL, AGRIC, COLE, VOL. 11. HARES ere Without Liming With Caustic Lime With Calcium Carbonate. Original Soil Ori (No Manure & No Lime). (With } Rike Ke ake Original Soil rig aaa Burnt Soll Burnt Soil (No Manure & No Lime). (With — na 7 ] = \) = i S s w= a > s 3 = = Original Soil I Original ‘Soil No Manure & No Lime), (With Manure) L. AGRIC. COLL. 10! a On Organic Compounds of Phosphoric Acid in the Soil. 1B K. Aso. It is a well known fact that peaty soils show much more phosphoric acid soluble in hydrochloric acid after they are ignited than before ignition. Eggertz! showed that ‘Mullkorper’ contained 0.15—7.58 % phosphorus and 0.55—2.09 % sulphur. The same author and JVz/son? pointed out that, in the hydrochloric acid extract of the ignited muck soils, a larger quantity of sulphuric and phosphoric acid is found than in the extract of the original soil by hydrochloric acid, hence it is inferred that phosphorus and sulphur are present in humus in form of an organic combination. Recently, Schméger*® made similar observations and concluded that a certain amouut of phosphoric acid is present in such soils in the form of nuclein. The fact that the soil of our college farm in Komaba contains nearly 11 9% of humus of a distinctly acid reaction and that hot concentrated hydrochloric acid does extract about double the amount of sulphuric and phosphoric acid than cold concentrated hydrochloric acid, as O. Ael/ner has observed years ago, induced me to search in what form a part of the phosphoric acid would be present. The soil serving for my investigation came from the uncultivated part of our fields and gave 27.16 % loss on ignition. It contained 10.9 % humus and 0.65 % total nitrogen in the dry matter. The soil was collected from several holes of a suitable depth after removing the surface layer, spread on mats and dried in the sun. The air-dry soil was then sifted 2 Biedermann’s Centr. Bl. f. Agric. Chem. XVIII, 1889. 2 Ibid. 3 Landw. Jahrb. XXV, 1896, and XXVI. 1897. 278 K. Aso. through a sieve of 1 mm. meshes. The samples serving for my in- vestigations were: J. Raw soil. II. Ignited soil. III. Soil steamed for three hours under pressure in Koch’s steam apparatus commonly used for sterilization. VI. Soil steamed under the pressure of three atomospheres in an autoclave for three hours. I. The raw soil. a). 50 grams of airdry soil were treated with too c.c. of cold hydrochloric acid of sp. gr. 1.15 at 15°C and digested for twenty four hours at the ordinary temperature shaking it from time to time, and then adding 100 c.c. water. 100 c.c. of the filtrate were evaporated to dryness on the water bath. After separating the silicic acid in the usual way with strong hydrochloric acid, the acid solution was filtered and filled in a measure flask of 300 c.c. and 50 or 100 c.c. of this solution were taken for deter- mination of sulphuric or phosphoric acid. Sulphuric acid was determined n the usual way with barium chlorid, and phosphoric acid was determined by the molybdic method, replacing first hydrochloric acid by nitric: acid. b). The quantity of the sample taken and the treatment with hydrochloric acid were quite the same as with a), but after filtering, the residue was washed thoroughly on a filter with water until a few drops of the filtrate did no longer show any reaction with silver nitrate. After evaporating the filtrate to dryness on the water bath, the after treatment was carried on in the manner described above in a). The result is seen from the following table, where the amount of dry matter corresponding to the solution used is stated :! 1 Of course, hygroscopic water of the soil was taken into account for the calculation of volume taken in the case of a), tO NI \O On Urganic Compounds of Phosphoric Acid in the Soil. PHOSPHORIC ACID. Dry matter Mg, P, O, P, O, found. EO- In average. used. found. gram, gram, gram. % % a. 6.4691 0.0147 0.0094. 0-145 ee 0.1445 b. 13.5204 0.0305 0.0195 0.144 SULPHURIC ACID. Dry matter BaSO, S O, found. 510, In average. used, found, gram. gram, gram. % % a. 6.46901 0.0372 0.0128 0.198 0.200 b. 6.4691 0.0378 0.0130 | 0.201 LT, The tgnited sotl. 50 grams of airdry soil were dried and ignited in a platinum dish, all precaution being taken to avoid loss, then the heat was raised gradually to a faint redness, stirring the soil from time to time with a platinum wire, until all organic matter was destroyed. The product thus obtained, possessed a reddish colour somewhat similar to that of oxide of iron. After keeping for a few days in the room, this soil was treated with 100 c.c. of cold hydrochloric acid and digested for twenty four hours at the ordinary temperature etc, as just described in the case of I. b.. Here, the treatment was quite the same in both cases, viz., a) and b). The following table shows the analytical data, where the quantity of dry matter stated corresponds to the solution used : 280 K. Aso. PHOSPHORIC ACID. Dry matter Mg, PO; P, O, found. In average, used, found. gram, gram, gram. % a. | 13.5204 0.0692 0.0441 oo 0.327 b. | 6.76¢2 0.0348 0.0222 SULPHURIC ACID, In average. Dry matter used, BaSO, found. S O, found. gram, gram, gram- % a. | 13.5204 0.1278 0.0439 0.325 13.5204 0.1280 0.0440 Ill. The soil steamed in Koch’s steam apparatus. 50 grams of airdry soil were thoroughly mixed with 100 c.c. of water in a beaker and steamed in Koch’s steam apparatus for three hours, there- upon the steamed soil was removed to a porcelain dish, and dried on the water bath. This sample after being exposed to the air for a few days was treated as those before, with the following result: PHOSPHORIC ACID. Dry matter Mg, P, O, P, O, found, In average. used, found. gram, gram, gram, % a, 13.5204 0.0316 0.0201 gtr (is ale 0.147 13.5204 0.0310 On Organic Compoundsiof Phosphoric Acid in the Soil. 281 SULPHURIC ACID. Dry matter Ba SO, S O, found. » O3 In average. used. found, gram. gram. gram, % % a. 13.5204 0.0888 0.0305 0.226 0.2255 b. 13.5204 0.0886 0.0304. 0.225 TV. The soil steamed in an autoclave. 50 grams of airdry soil were put in a tall beaker, mixed with 100 cc. of water and placed in an autoclave containing some water, then heated until the pressure increased to three atomospheres and at that pressure the steaming was continued for three hours. The steamed soil was trans- ferred into a porcelain basin, dried, and also left like the other samples exposed to the air for a few days. Phosphoric and sulphuric acid were determined by extraction with cold hydrochloric acid of sp. gr. 1.15 at 15°C as described before. The result obtained here was as follows: PHOSPHORIC ACID. Dry matter used, Mg, P, O, P, O, found, found, Pe Oe In average. gram. gram. gram. % % a. 13.5204 0.0477 tj Se Se SE eee ee en 0.351 13.5204 0.0236 SULPHURIC ACID. Dry matter BaS QO, SO, found. SO; | In average. used. found. gram. gram, gram, % % | a. 6.7602 0.0480 0.0365 0.244 } 0.245 6.7602 0.0485 0.0166 0.246 282 K. Aso. The results here obtained are seen at a glance in the following table : The raw The ignited The soil steamed | The soil steamed Treatment. in Koch’s steam in soil. soil. apparatus, an autoclave. Percentage of ae it O.1 0.32 OI ae Phosphoric acid, 45 g2f oI 0-351 Percentage of 0.200 0.325 0.226 0.245 Sulphuric acid. The amounts of phosphoric acid and of sulphuric acid obtained by extraction of the orégznal soil with cold hydrochloric acid corresponds to those pre-existing as phosphates and sulphates respectively, while the differences between the percentages of those and of the phosphoric and sulphuric -acid found in the zgzted soil, might be derived from certain organic compounds. The soil steamed in Koch’s apparatus did not liberate any considerable quantity of sulphates and phosphates, but the autoclave treatment had a better effect. These results are in favor of Schméger’s inference above mentioned. In the following table, a comparison of Schmégers soil and ours is given: Raw Sol. ew PHO 50; Poe SO; Sedliner2 moor. 0.123 0.134 0.246 0.244 Soil from Komaba. 0.145 0.200 0.351 0.245 In cold hydrochloric In hot hydrochloric extract. extract, Soil from Komaba.? 0.19 O.11 0.34 0.20 * Landw, Jahrb. Bd. XXV. 1896. ‘This soil was analysed by Schméger. 2 Landw, Versuchst. Bd. XXX. This soil was analysed by Kellner, On boiling with hydro- chloric acid, nuclein in the soil might be decomposed. On Organic Compounds of Phosphoric Acid in the Soil. 283 Schméger’ inferred that lecithin was not present in his soils. I made some experiments about this point with the soil of our field. Several hundred grams of the soil, after well-drying, were extracted first with ether and then with alcohol and the two liquids were united and evaporated on the water bath. A part of the evaporation-residue was heated with a mixture of sodium carbonate and some potassium nitrate, and the ash dissolved in nitric acid and tested for the presence of phosphoric acid therein with ammonium molybdate and the characteristic yellow colour was produced.?° The presence of lecithin in the soil became thus very probable, hence I made a quantitative determination according to the method rccommended by F. Schulze.* The result is shown in the following : Dry matter Ethereal and Ma, Bs.0, F.O,.tound, Lecithin aes used. alcoholic extract.| found, found. gram. gram, eram. gram. gram. % 40.561 0.0616 0.0025 0.0016 0.0182 0.004 40.561 0.0535 _ 0,0030 0.0019 0.0218 0.005 In average 0.05755 0.00275 0.00175 0.0200 0.0045 In roo parts of dry soil, 0.0493 parts of lecithin were found, so that 100 parts of humus contained 0.452 parts of it. The ethereal extract of this soil was dark greenish brown. The coloring matter might be due to a partial decomposition product of the chlorophyll. The quantity of lecithin _is but small so that it can not be taken into serious consideration. To see how much phosphoric acid may be rendered available by destroying organic matters in this soil, I determined the phosphoric acid soluble in 1% citric acid4 in the following way: 4 Landw. Jahrb. Bd. XXV._ 1896. 2 Besides, I made a qualitative test by decomposing the lecithin with baryta and preparing the double chloride of cholin and platinum. 3 Chemiker Zeitg, 1897. 4 Dyer: Jour. Chem. Soc. Vol. LXV. 1894. 284 K. Aso. a) 81.855 grams of dried soil were digested with 200 c.c. of 1% citric acid for forty eight hours at the ordinary temperature, shaking it from time to time. The filtrate and wash water was evaporated, and after separating silica, phosphoric acid was determined by the molybdic method in the usual way. b) 81.855 grams of dried soil were ignited in a platinum dish and the extraction with 1% citric acid and after-treatment was quite the same as in the case ofa). The result will be seen in the following : Dry matter used. Mg, P, O, found, P, O, found. P1Oz gram. | gram. gram, % Tgnited soil. 40.927 0.007 I 0.0045 0.011 Raw soil, 40.927 0.0035 0.0022 0.005 It is clearly shown that the quantity of phosphoric acid soluble in 19% citric acid in the soil can be increased considerably by destroying organic matters. Summary. 1. Phosphoric acid is present in humus soil in organic and anorganic forms. 2. The chief organic phosphoric compound is nuclein. Besides, a very small quantity of lecithin is present. Both compounds can be partially due to the bacterial flora of the soil, partially to the decaying plant roots. 3. The phosphoric acid in the organic compounds become available by burning the humus soil. I determined also phosphoric acid contained in Matiere noire obtained from this soil: 0.249% I’, O. was contained in Matiere noire, in the dry soil, Of course, this includes some P, O, of aluminium phosphate, because aluminium phosphate is somewhat soluble in ammonia. P, O, from raw soil (0.145°%)+P,, O; from matiere noire (0.249%) =0.394.%, while P, O, frem the soil steamed ee ‘ 0/ under 3 atm, =0.351%. On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. BY IM. Nagaoka. The question of the protein formation in plants has been the subject of numerous investigations during the last fifty years in Europe by various au- thors, as Hartig, Borodin, Pfeffer, Kellner, and especially E. Schulze and his students, and in our college O. Loew, Y. Kinoshita and U. Suzuki made also various observations in this line. It was further known long since that the nitrogen of nitrates as well as of ammoniacal compounds can be utilized for the formation of asparagin and protein. In the practical and economical point of view, the manurial effect of the nitrogen in both, nitrates and ammonium compounds, has also been compared by many agricultural chemists with different crops and also in different soils, but the results did not always agree. In certain respects, even contradictory statements have been published by different observers, due to the different conditions of soil. Aowssingault was the first to recognize the high manurial value of chili saltpetre. Since then Barley, Ff. Bennet, Lowel, Kuhlinann, P. Wagner, Maercker and others have carried out extensive experiments in regard to the efficacy of these salts in comparison with other nitrogenous manures and the valuable state- ments of these investigators have rendered the Chili-saltpetre very popular as a manure. In 1896, however, Paguaul! published the results of his experiments on the assimilability of nitric and ammoniacal nitrogen with several agricultural 2 Ann, Agronomique 22 (1896). 286 M. Nagaoka. plants and inferred that the ammonium sulphate was decidedly superior to the nitrates in all his experimental crops. . Klopfer! observed also from his experiments that in presence of other necessary untriments the ammo- nium sulphate is more beneficial than the sodium nitrate and he expressed also his conviction that sulphate of ammonia is more economical than nitrate of soda, especially for application to cereals in the spring; but soon after- wards P. Wagner? inferred from his own experiments that K/opfer’s obser- vations were erroneous. H, F. Adams* has also made some observations regarding the relative assimilability of the various forms of nitrogen upon an acid soil limed and unlimed arrived at the following conclusions: 1. On a very acid soil, ammonium sulphate has worked like a poison instead of an effective fertilizer. 2. Where air slaked lime was applied with ammonium sulphate the nitrogen proved nearly as valuable as similar quantities in form of sodium nitrate. In 1901, 1. Bachmann compared the effect of sodium nitrate to ammo- nium sulphate upon potatoes, fodder beets, and kohl-rahi; in which experi- ments the sulphate proved apparentiy more effective than the nitrate. The experiments, which were performed on barley, at the Japanese agriculture experiment stations proved the value of nitric and ammoniacal nitrogen to be varying according to the nature of the soils experimented on, while in the results obtained by S. Machida® on the manuring experiments of Poly- gonum tinctorium, the value of both, nitric and ammoniacal nitrogen, was estimated as to be almost the same. Although the relative effects of nitric and ammoniacal nitrogen have so often been made an object of practical investigations with defferent crops * Jahresbericht f, Agri. Chemie 1899 s.s, 107-109 and Deutsch, Landw, Presse Bd. 25 (1898) No. 25 pP.p. 271. Dentsch, Landw, Presse Bd. 25 (1898) No, 30 p.p. Res tw Rhode Island Station report 1897 p.p. 241. - Frihling’s Landw, Ztg. 50 (1901) No 11 p.p. 386-387. ® Report of the Imperial Japanese Experiment Station No 21. On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 287 and on different dry fields, yet their values are not yet certainly fixed for all kinds of soils, whilst, as to the nutritive effect of these two forms of ni- trogen towards aqueous plants, especially to paddy rice, practical experi- ments, as far as I know, are entirely wanting. However, as a whole, in all irrigated soils the so-called process of ni- trification does not generally take place and ammonia is, in such soils, always the result of the putrefaction of some nitrogenous organic matters, hence rice plants growing in such conditions of soils might generally be supposed to be provided with ammoniacal nitrogen alone. Hence the in- vestigation on the behavior to nitric nitrogen might seem to have no special importance. However when the richness, in nitric nitrogen, of some manures which are often used in the cultivation of rice plants are considered and also the occurrence of a slight nitrification in the common paddy fields during their exposure to the air in the late spring and in a nearly dry state, the question will certainly have some interest from the scientific as will as the practical point of view, especially for the cultivation of upland rice plants, which is generally cultivated in dry fields in which nitrification always largely takes place. I have carried out several experiments, to study whether nitric nitrogen can be utilized as well as ammonium nitrogen by rice plants under different conditions. Before entering upon the details. I must express my earnest thanks to the professors Loew, Kozai and Toyonaga, my colleagues in the college, who kindly overlooked and revised this and also my former papers; I must also pay the same thanks to J. Sudsuki and T. Koyasu, the assistants in our laboratory, who rendered me valuable assistance in my work. 1. Lxpertments on-the effects of different nitrates as compared to ammonium sulphate, upon rice plants. These experiments were carried out, in 1899, with 92 porcelain pots each measuring 25 centimetres in diametre and performed in the new glass house of our college. The principal questions, I had in mind, were: 1. How behave different nitrates and ammonium sulphate upon rice plants? 288 M. Nagaoka. 2. At what period of the plants growth, the nitrogen is most assi- milated. What doses of the nitric and ammoniacal nitrogen are profitable to Oo the plants. The soil was taken from one of our experimental fields which had been cultivated without manures for 5 years. After the field was well ploughed, it was irrigated and stirred to a condition of mud, and in order to get it free from all rootlets of the preceding crops, the mud was carefully sifted. It was then exposed to the air for three days. Before the application of the thus prepared soil, all pots received about 4 kilograms of pebbles adjusting each pot to 10 kilograms in weight where- upon each received just 11 kilograms of the soil; the total weight of each pot then being 21 kilograms. The nitrogen manures were the pure nitrate of sodium, potassium, calcium, barium, strontium and magnesium, besides the ammonium sulphate. It was further proved by careful tests that all the nitrates were free from nitrite and perchlorate and the ammonium sulphate from sulphite and cyanide. On July, 14 (1899) as the general manure, the phosphoric acid in the form of sodium phosphate and the potash as potassium carbonate were applied at the rate of 100 kilograms respectively per hectare. The various forms of the nitrogen were added on the 15th; the quantities of the nitro- gen applied per hectare and per pot, are given in the following table :— On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 289 Nitrogen applied Nitrogen applied Kind of the nitrogenous per ha, per pot. No of pots. manures, Kg. Gr, sor, | Without od E 2, 33; 63 18) 12) nitrogen. 3, 34 64 fo) ° 4, 355 65. 25 0.122 = 36, 66. Ammonium 50 0.2454 6, vi Peasy sulphate. 50 0.2454 7 38, 68 50 0.2454 8, 39, 69. 25 0.1227 9, 40, =o. Sodium 50 0.2454 10; 41; Tile nftrate. 50 0.2454 it,5 —A2; 72 50 0.2454 12, 43; 73- 25 0.1227 13, 44, 74. Potassium 50 0.2454 ely 455 75- nitrate, 50 0.2454 15, 46, 76. 50 0.2454 TOM 47s ie 25 0.122 175 48, 78. Calcium 50 0.2454 eee) 19: nitrate, 50 0.2454 19, 59, 8o. 50 0.2454 20, ie 81. 25 0.1227 21, 52; 82. Barium 50 0.2454 22, 53, 83. nitrate. 5° 0.2454 23, 54 &4. 50 0.2454 24, 55> 85. 25 0.1227 25, 56, 86. Strontium 50 0.2454 26, 57, 87. nitrate. 50 GAGS 27; 58, 88. 50 0.2454 28, 59, 89 25 0.1227 29, 60, go. Magnesium 50 0.2454 30, 41, gl. nitrate, a? O28s% 290 M. Nagaoka. On the 16th, after addition of 2000 c.c. of distilled water to each pot the specially nourished young rice plants (variety Satsuma) 65 days old were transplanted to all pots, each pot receiving 3 bundles of six healthy equally nourished individuals. All pots were carefully weighed every day up to the middle of the blooming of the plants and the losses of weight owed to the water evapora- tion from the pots and to the perspiration of the plants were daily replaced by distilled water. The chief difference observed was that the plants which received tlie ammonium sulphate exhibited a normal green colour and appeared quite healthy and further, they showed a vigorous tillering while the plants sup- plied with nitrates were yellowish and their tillering was very faible but the seeds ripened about 10 days sooner than those of the former plants. The pots of the small doses of nitrogen and the last series of the large doses of the respective experiments were left until the plants were in a state of full ripeness while the other two series of the large doses were harvested in two periods, the first cutting being taken on the 15th of August and the second on September 15th. The following table contains the yields per pot, the averages of the three parallel trials and the contents of nitrogen given by the chemical analyses! of the crops. 1 The determination of the nitrogen of these and the following experiments were performed after the common method of Kjeldahl, hence the results of the determinations do not contain the nitrogen of nitric acid in the harvested crops, On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 291 First crop. Second crop. No | Yield | , erage [N-inthe Bead chee No | Yield |, crage|N-inthe ae per pot. total crop eee af: per pot. jtotal crop per pot. _ per pot. pots. gr. gr. gr. pots, er, gr. gr, 2 15.75 I 6.12 Without 6.14 0.093 33 16.04 16.60 | 0.141 32 6.16 nitrogen, 63 18.01 5 10.25 6 22.92 Ammonium 36 9.63 10.01 0.159 37 23.01 23.18 0.186 sulphate, 66 10.16 67 23.61 9 6.36 10 16.74 Sodium 40 5°85 6.15 0.094 41 17.16 16.85 0.147 nitrate. 70 6.25 71 16.65 13 8.75 14 18.19 ‘ potassium 44 6.76 7.23 0.100 45 16.75 77 0.146 nitrate. 74 6.16 75 16.56 17 7.51 18 15.95 Calcium 48 8.85 7.06 | 0.106 49 17.43 | 17.16 | 0.136 nitrate, 78 7-52 79 1S.11 21 8.03 22 18.06 Barium 52 8,02 8.15 0.099 Be 17.2 18.06 | 0.154 nitrate, 82 8.40 83 18.88 25 7:25 26 17.81 Strontium 56 8.46 7.82 | 0.0997 57 18.44 | 17.58 | 0.143 nitrate, 86 7:75 87 16.48 29 8.31 30 16.94 Magnesium 60 6.77 7.01 0.102 61 17.51 17.11 0.159 nitrate. go 5.95 | gl 16.88 7.39 0.1001 || Average of the nitrates. | 17.22 0.1475 292 M. Nagaoka. From the above two series of the results, it will be noticed that all ni- trates had but little influence upon the organic matter production of the rice plants in both early periods of vegetation, while the ammonium sulphate caused a most prominent augmentation of the yield. Moreover the actual quantity of the absorbed nitrogen from the various nitrates was in all cases, equally trifling. There were no essential differences between the effects of the various nitrates. When the actual surplus of the yield and also the surplus of the assimi- lated nitrogen of the ammonium sulphate over the pots not supplied with nitrogen, are assumed respectively, in both periods, to be 100, the ratio of the surplus yield and also the ratio of the surplus nitrogen assimilability of the nitrates, calculated from their average results, will be as follows :-— Ratio of the surplus Ratio of the surplus nitrogen yield. assimilability. Ammonium : Ammonium . sulphate. Nitrates, sulphate, Nitrates. In the first period, 100 32.3 100 10.8 In the second period. 100 10.9 100 14.2 Thus it will be observed that the nitrogen of the nitrates was, in both periods, utilized by the plants in a considerable less degree than that of the ammonium sulphate and consequently the yields in all the pots supplied with the various nitrates did not reach the harvest yielded by ammonium sulphate. If the average of the above two series of figures may represent the rela- tive action between the sulphate and nitrates, the latter action will be, in the first period, 21.6 and in the second period 12.6 to the 100 of the former. At last, 1 give, in the following two forms of table, the figures obtained from the harvests, which have been cut on November the 11th, from the pots supplied with two different doses of nitrogen that is to say 25 kilograms and 50 kilograms per hectare. On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 293 TABLE; A. Results obtained from the pots which were supplied with nitrogen at the rate of 25 Kg. per hectare. 59 Magnesium nitrate. é 14.54 16.77 16.19 32.66 Shea or Prine: Average per pot. Nitrogen No of. Kind of nitrogen Persson we in the Straw. | Grains. | Total total crop pots. manures. crop. per pot. gr gl gr. gr gr 3 16.33 34.5 Without nitrogen. ‘ jaz 15.80 17.02 32.82 0.2078 64 17.61 4 19.86 35 Ammonium sulphate, 5 22.70 21.39 19.74 41.13 0.3030 65 16.67 8 17.78 39 Sodium nitrate. ; 16.78 16.51 17.05 33.56 0.2082 69 17.14 12 16.44 43 Potassium nitrate. : 16.29 | 16:29 | 16.4% | 32.70 0.2079 73 16.50 16 16.70 47 Calcium nitrate. 3 Wye5t 18.04 17.49 35-53 0.2089 77 18.25 20 16.20 51 Barium nitrate. y 16.93 16.21 16.75 32.96 0.2086 81 17.12 24 16.65 55 Strontium nitrate, ‘ 14.92 16.11 15.92 32.03 0.2081 85 16.20 28 ; : 17.30 2904 M. Nagaoka. TABLE B. Results obtained from the pots which were supplied with nitrogen at the rate of 50 Kg. per hectare. Siraws SoG: Average per pot. Nitrogen No of Kind of nitrogen naw) GRAMS), in the Straw. | Grains. | Total | total crop pots. manures, crop. per pot. gr gr. gr gr. 7 22.41 22.21 38 Ammonium sulphate. 21.19 21-52 22.17 0.3040 68 22.91 23.27 61 17.37 | 18.05 42 Sodium nitrate. 15.60 | 1662 16.53 0.2081 72 15.98 16.65 15 18.63 16.75 46 Potassium nitrate. 17.10 17.51 17.41 0.2086 76 16.49 17.43 19 16.01 17.39 50 Calcium nitrate, 16.63 17.41 16.75 0.2085 so 17.62 18.32 23 16.26 17.45 54 Jarium nitrate. 16.02 | 17.10 | 16.70 0.2084 84 17.83 15.76 27 16.40 18.27 58 Strontium nitrate. 18.04 17.62 16.74 0.2085 88 15-77 17-39 a1 16.50 17.21 62 Magnesium nitrate, 17.69 18.19 | 16.79 0.2086 From the figures of the above two tables, we may learn that the small doses of the various nitrates (25 Kg. N. per hectare) had almost no influence ~ On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 295 upon the rice production whilst that of the ammonium sulphate was more favorable. On the other hand, from the larger doses (50 Kg. nitrogen per hectare) of the various nitrates a little more nitrogen was absorbed by the plants than in the cases of the smaller doses leading to an insignificant augmenta- tion when compared to the yield of the pots not manured with any nitrogen ; but as these increases varie within a very narrow limit, it is not worth at- taching a practical importance. The larger dose of the ammonium sulphate had again a better action than that of the nitrates, yet when its increase in yield is compared to that of the smaller dose, its effect was not large, so that we can infer that the small dose of the sulphate was sufficient for the plants. In 1896, W. Schneidewind' has drawn the following conclusions from his experiments on the action of various nitrates upon the produce of oats: 1. Die aus den verschiedenen Nitraten aufgenommenen Stickstoffmen- gen waren vollkommen gleich. 2. Die Dingung mit Kaliumnitrat hat die grésten Strohmengen er- zeugt, im Kérnerertrag jedoch das geringste Gewicht. 3. Die Diingung mit Magnesium nitrat hatte die grésste Kornermenge erzeugt. With regard to the above statements, my present results agree only with his first conclusion and there were not, in fact, a difference noticeable between potassium nitrate and magnesium nitrate. However, in the preceeding tables there will be seen a peculiar pheno- menon in regard to the proportion of the straw and grain, which have been produced in nearly equal quantities (the quotient of yield being about 100) ; in some cases the weight of grains even excuded that of the straw. This had never been observed in our former expriments with paddy rice plants and also in my later experiments, this is the a most exceptional occurrence this phenomenon cannot be due to a special influence of the manuring but to 2 Jahresbericht f. Agric, Chemie. Bd, XX 1897. p.p. 228. 296 M. Nagaoka. other conditions of which the exceptional late transplantation! of the young rice plants must specially be remembered. As to the nitric nitrogen in all harvested plants, it was not quantitative- ly determined, but by the way of qualitative test with diphenylamin, it was shown that all plants which received the nitrates showed a distinct reaction for the nitric acid and the reaction was stronger in the earlier period than in the later, Tl, Experiments with different doses and different application of sodium nitrate aud ammonium sulphate to paddy rice plants. These experiments were carried out in the field in the same year as the former in the glass house. Fifty seven wooden frames, each representing an area of 0.826 square metre, were placed in a paddy field of our experimental farm. The soil was taken from the adjoining field which had not been manured for a long time ; the general treatment of the soil in the frames was not essentially different from that described in my former papers. The potash was applied, July 29 as potassium carbonate and the phosphoric acid as double superphosphate, at the rate of 100 kilograms per hectare respectively. The rate and actual quantity of the nitrogen, which was applied in the form of ammonium sulphate and sodium nitrate, per frame and also the mode of application are seen from following table. 1 Jn my experiments, the transplantion of the young rice plants was about 20 days later than in common practice, On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 297 I, 13, 14, frames. 20, or; 22, 23; 27; 29, Kind of Nitrogen Nitrogen Remarks applied | applied nitrogenous per ha. | per frame. | on the | manures. Kg, gr. | nitrogen application. Without nitrogen. fo) fe) fo) 25 2.083 The whole quantity was Ammonium 50 4.166 applied at the tran- sulphate. 75 6.249 splantation, 25 2.083 | 3 was applied at the tran- ; 50 4.166 | splanting and } after | ; 75 6.249 | 30 days. 25 2.083 3 4 was applied every 15 50 4.166 ” days. 75 6.249 | 25 2.083 The whole quantity was Sodium 50 4.166 applied at the time of nitrate, 75 6.249 | transplanting. 25 2.083 | 3 was applied at the " 50 4.166 | plantation and 2 after 75 6.249 30 days. 25 2,083 a 4 was applied at every 15 50 4.166 days. 75 6.249 On the 30th of June, the young rice plants well nourished and 50 days old were transplanted as usually. The crops were harvested on November 21 with the following results (the detailed figures of the yields per frame are seen from the last page). The last column of the table contains the figures of the total nitrogen con- tained in the total crop per frame, as found by chemical analysis. 298 M. Nagaoka. | N, applied 4| No Kind of per ha. “| Straw. | Full | Empty | Total | Nitrogen Kg. | grains. | grains. | crop. |in the total of nitrogenous crop per “4 Manner frame. frames. manures, of gr. gr. gr. gr, gr. application.| | if 2G, hag, o o o 303.9 | 266.3 4.8 575.0 6.866 By V2 AO; 25. given 388.9 334.5 4.9 723.6 8.521 | Ammonium | sayt 22k An 50. in 441.5 | 379.5 6.0 827.0 9.825 sulphate. 4, 23; 42: 75s) 1 doses 480.6 401.3 5-4 887.3 10.802 | | Bea eat Aas 25. given | 361.3 303.1 5-5 669.9 7.910 6, - 25, 44 ri | So.) sin 408.5 | 359.1 5.7 773.3 8.966 ” | 7, 26,0 45 PO Gigs, “AGIOEES 463.1 | 406.6 6.4 876.1 9.739 | |) 8, 27, 46. | as. given | 3481 | 316.3 4.9 669.3 7.591 ” | 5 | 9, 28, 47. | iP oe | 3989 | 347.6 | 5.3 | 7518 | 8.763 10, 29, 48. i. 75. 4 doses, | 406.3 | 3589 | 5.9 | 77n1 | 9.094 iE | ae Hits) 930 40..0)| ' | 25. given 353-1 307.5 4.9 665.5 8.005 Sodium IZFAUZ TS 150: 50, in 349-5 | 312.7 Ree 667.3 8.152 nitrate, 13s 32, ae 75. 1 dose; |) 36Gck 328.4 6.4 702.9 8.360 Ay gables 25, given 349.8 | 307.3 | 5.4 | 6625.| 7.347 15; 34, 53- é 50. in 347-2 | 309.8 5.6 662.6 7.616 Misr ee pee || a 75, 2 doses. | 3380 | 302.7 54 646.1 7.484 —— (> 75) 30,0 55: 25. given | 350.4 323.5 5:5 679.4 7.667 ” | 1Sj0 375) 150. 50. in | 323.2 287.6 5.1 615.9 6.807 19;°138; 57: Ra 95 4 doses. | 339.5 | 303.5 5-4 648.4 7.458 These results demostrate again an inferior action of the sodium nitrate upon the rice plants, as both the yield and assimilated nitrogen in all the frames with the nitrate in different doses varied but within a narrow limit ; whilst the nitrogen of the ammonium sulphate was easily assimilated in the proportion to the quantities applied, as seen from the increase of the yields. Since all paddy fields are irrigated from time to time during the whole On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 299 rice vegetation and since they are thoroughly covered with water, thus re- ducing processes (denitrification) being enhanced, losses of nitrates may occur in two ways. Hence it might be deduced that the application of nitrates in several periods of the plants’ growth would secure better results than the applica- tion in one dose ; but my results sufficiently show that no better effect can thus be gained. In such paddy field as our’s, the late and divided dressing with nitrate or ammonium sulphate to rice plants has no advantage whatever. Since these experiments were performed more in accordance with the actual practice than the former experiments, it will be of some value to compare the manurial value of the sodium nitrate to that of the ammo- nium sulphate. Thus when the surplus yield (2.52 grams) caused by the medium dose (applied. at once) of the ammonium sulphate over the frames without nitrogen, is assumed to be 100, the relative increase caused by the same dose (applied also at once) of the sodium nitrate will be 36.6.. Again, the actual percentage assimilability of the sulphate nitrogen is estimated as much as 719 while that of the nitrate is only 30.9%, hence when the former percentage number (71) to be assumed as 100 the relative assimilability of the nitrate will be 43.5. The average of the relative increase and assimi- lability constitutes its manurial value thus, 36.6435 14.6, Hence it follows that the manurial value of the ammonium sulphate is 2.5 times greater than that of the sodium nitrate for paddy rice cultivation. In order to determine the after-effects of the residual nitrogen of both, ammonium sulphate and sodium nitrate, in this second series, the soils of all the frames, having been untouched during the coming winter, were, in the next season (1900) cultivated again for the same species of rice plants and with fresh application of the general manures as in the preceeding year, but without further addition of ammonium sulphate or sodium nitrate. The transplantation of the young rice plants took place on the 28th of June, nad the 300 M. Nagaoka. crop were cut on the 11th of November. Here, the details in yields per frame being likewise left to the last pages, we shall in the following table only give the averages of the three equally treated frames. No Kind of N, applied Straw, | Full Empty | Total | grains, grains, crop. of nitrogenous per ha. | frames, manures, Keg, gr. or, er, | gr. ly, $210) 210 Without N, oO 343.0 205.3 3.4 | 641.7 | BFE, AO . 25 37,12 326.6 4:9 "= =JO27 Ammonium 3, 22, 41 50 370-3 330-3 36 | 704.2 sulphate, | 4, 23, 42 75 411.3 359-0 4-4 | 774-7 he HL alee 25); 370.7 339.3 3.6 713.6 6, 25, 44 a 50. 361.1 B127, 4.4 678.2 7, 20, Ak. vac 400.0 356.3 6.2 762.5 8, 27, 46. 25. 338-8 |) 315-7 82 9. Aare 9; 28, 47. ; 50. 377-7 3077 34 698.8 Io, 29, 48. hen YS 403.7 336.3 4.2 744.2 IT, 30, 49. 25; 348.0 | 295.0 4.6 | 647.6 Sodium ie Boe fo, 50 355-7 322.3 3.6 681.6 nitrate, 13 59 G28 eG tse 367.5 357.0 3.6 708.1 — : al et ee 14, 33, 52. 25 | 366.0 322.0 533 691.3 ” | 15, 34, 53 | 50. | 374-7 316.3, 32 694.2 16, 35, 54- 75. 347.0 295-7 4.4 647.7 17, 36, 55. 26, 360.8 307.0; je eo 672.7 185) hye) 50s 50. 364.0 336.3 4.3 704.6 19, 33, 57. 75+ 379.0 337.0 3-9 719.9 | . . . By a superficial glance at the above figures, it might seem that the re- | sidual nitrogen displayed some special good influence upon the rice produc- tion. Tlowever, when we take into considration the high proportion of the total crop also in the frames not supplied with any nitrogen, which frames On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 301 giving in average 66 grams more yield than in the first year, the general augmentation of the yields in all cases can not be due to the efficacy of the residual nitrogen but to other causes very probably to the highly favorable weather which prevailed in 1900. As to the actual effects of the residual nitrogen, it is seen, as a whole, that, although the residues of the ammonium sulphate had again somewhat better influence than those of the sodium nitrate, yet in both cases the effects were far behind those when nitrogen compounds were freshly applied. I give in the following table, the actual surplus yields obtained in the first and second year respectively. | : = ied i 5 Sur ri r No of | Kind of | N. applied in the | N. left for the urplus yield. er. hitradenods Ist year per ha, second year ‘ $ per frame. Aue : : rames, manures, Kg or First year. {| Second year, | § | | De WIT «40; | 25. given 0.428 148.6 61.0 Ammonium ; | Bell. AN. | 50. in 1.207 252.0 62.5 sulphate. | Arent 42 | 75. one dose, 2.312 3123 133-0 | 5, 14, 43. | 25- given 0439 | 949 | 71.9 39 } } Gyercy 44: | Foy al 2.066 198.3 136.5 | ” : TeelOy 45 75. 2 doses 3.378 301.1 120.8 S17; 46. 25. given 1.358 94.3 16.0 9 Oy 1c; 47 5Oge Um 2.269 176.8 57.1 ” | | 10, 19, 48 75. 4 doses, | 4.021 1098.1 102.5 ly20,, ‘40: a |} 25. given 0.944 90.5 6.1 Sodium Hed, ts | 1toF 5 | 5o. one 2.880 92.3 39.9 nitrate, Koos Sh l- 75- dose. 4.755 | 127.9 66.4 14, 23, 52. 25. given 1.602 $7.5 48.6 | ' | % | ; : es Tip 2450853: SOs an 3-410 $7.6 52.5 ” | | s | Gee 254 S54. 75. 2 doses. 5.631 | ye 6.0 Es 420;, 55 25. given 1.282 104.4 31.0 ” Toye 27, 56 Xm pin -——— 40.9 62.0 195 928. 57. | i -¥5.. 04 doses: 5.657 73.4 78.2 on EE Ee ee, eee EE eee 302 M,. Nagaoka. Thus in the second year, the residual nitrogen of both, ammonium sul- phate and sodium nitrate, had more or less action upon the rice plants yet, excepting but the residual nitrogen of the largest dose (last series of the nitrate) of the nitrate, there is no case in which their actions exceeded or attained an equal rate to that of the first year, although, as proved by the results of the chemical analysis, there were left still sufficient quantities of the nitrogen to produce crops similar to those of the first year. As to the residues of the nitrogen after late application there was more left in the soil than in the case of early application, yet there was no better effect in the former case. This may be due most probably to the fact that the nitrogen which was applied at the early period, was relatively more absorbed by the plants and a part of the assimilated nitrogen was reserved in the roots which by de- composition in the following year produced comparatively some good effect upon the second crops; on the other hand, the nitrogen which was applied later, having been less absorbed by the plants, was much washed away by irrigation in both first and second year, and consequently a certain quantity was lost. Appendix. YIELD PER FRAME (rst year). 7 soot | Se | Ea Bey oe | frames, gr, gr, er, | frames, er, er, gr, a = ee a ee ; 1 314.0 268.5 4.6 | 3 433.8 383.5 55 20 301.3 252.5 5.1 22 413.8 345.0 6.4 29 296.5 278.0 4.7 41 476.8 410.0 6.2 es isgaed 2 397:2 342.0 2 4 491.3 395.0 7:0 21 393-3 345.0 49 | 23 483.3 407.0 6.0 40 362.2 316.5 5.5 42 467.3 402.0 3.1 DS we On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. frames. | or, | er. er. frames, | er, gr, | 5 | 342.5 | 274.0 | 2 13 351.8 298.5 | 24 328.3 289.0 | 5°55 32 241.3 302.5 | 43 413.0 346.0 57 | 51 411.3 | 384.2 | 6 | 351.3 204.2 | Ise) | 14 Biles 263.0 25 434.8 374.5 Gs ot 33 368.8 315.0 44 439-3 408.7 5-5 52 369-3 344.0 7 495.8 418.2 6.5 | 15 207.3 258.0 26 456.3 413.0 5.4 | 34 346.0 308.5 45 437-3 388.5 73 | 53 398.3 363.0 8 391.0 355-0 5:3 | 16 277°3 253-5 27 334-0 309-5 4.3 35 341.5 301.5 46 319-3 284.5 5.2 54 395-3 353.0 .9 3715 317.0 6.8 | 17 323.8 300.5 28 443.3 375.2 4.8 36 308.5 272.0 47 381.8 350.5 4.4 | 55 418.8 398.0 | | 10 390.8 a6 i.2 5.0 18 328.3 278.5 | 29 408.8 366.2 56 | 37 257.5 206.0 | 48 419.2 359.4 7.0 | 56 383.8 378.3 | II 331.0 279.7 2. / 19 339-3 309.0 | 30 323.3 2777 6.0 38 339.5 301.0 | 49 405.0 365.0 5.9 | 57 339.8 300.5 | 12 346.8 310.0 4.3 | | | 31 326.8 284.0 6.2 | 50 375.0 344.2 4.9 | | | Empty grains, = gr. 7.0 304. M. Nagaoka. YIELDS PER FRAME (Second year). nS eee Straw. Full Empty | Straw. Full Empty ; No of grains, grains, | Noof grains, grains. frames, or, gr, gr, | frames. gr, er. er. I I 353-0 282.0 3.9 | 9 396.0 311.0 3.2 20 335.0 294.0 3 28 383.0 346.c 3.0 39 336.0 310.0 27 | 47 354.0 296.0 4.4 2 362.9 308.0 | 5:7 | 10 374.0 313.0 4.2 21 420.8 380.0 2 | 29 4130 366.0 3.0 40 330.0 291.8 4.9 | 48 424.0 330.0 a3 ee ee eee eee ceaal aes eee 3 368.0 325.0 4.2 ! II 358.0 278.0 5.1 22 388.0 351.0 34 30 28210 295.0 2 41 355.0 315.0 3.0 | 49 354.0 312.0 35 ee | ee | LM <..- °° 4 457.0 379.0 5.0 12 331.0 303.0 4.2 23 392.0 349.0 37 |) 3 379.0 330.0 2.7 42 385.0 349.0 4.5 ite) 357.0 325.0 - 4.0 ee ee —_—, bE ae 5 370.0 328.0 3-9 i 13 372.0 331.0 3.2 24 411.0 3725 39 | 32 373.0 359.0 3.9 43 331.0 317.5 3.0 | 51 357-5 321.0 3.7 6 371.0 277.5 6.0 | 14 383.0 357.0 3.0 25 364.5 316.5 3.5 33 350.0 286.0 Be 44 348.0 344.0 3-7 | 52 365.0 3230 3-7 7 439.5 381.5 6.2 | 15 388.0 302.0 4.0 26 368.5 346.5 50 |, 34 374.0 334.0 2.5 45 392.0 341.0 7.4 53 362.0 313.0 oie 8 315.5 271.5 3.5 | 16 358.0 276.0 5.2 27 327.0 312.5 3.0 15 331.0 297.0 2.4 46 374.0 363.0 3.2 | 54 352.0 314.0 5:5 On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 305 | ee a en | ea frames. | gr er. | gr frames, | en | er, Pa a cae ae 17 349.0 276.0 | 6.5 19 384.0 348.0 | 3-0 36 | 365.5 3425 | 30 Be eset sazo | 32 55 | a e | 5.2 57 | 370.0 316.0 5.5 18 341.0 302.0 | 4.5 | a7 378.0 348.0 | 2:2 | | | | 359-9 5.2 Ill, How do Sodium nitrate and Ammonium sulphate act on Upland rice, compared with Paddy rice ? As upland rice is always grown in ordinary dry fields and under quite different conditions as paddy rice it was of interest to compare also here the effects of nitric and ammonical nitrogen. I paid attention to the following principal questions: 1. How and how much can nitric nitrogen be utilized by upland rice plants under ordinary conditions (that is to say without irrigation) ? bo How is the behavior under conditions similar to those of paddy rice cultivation (with irrigation) ? 3. What difference can be observed between the upland rice plants and paddy rice plants when the latter are cultivated at the same time under just the same condition as the former plants ? These experiments were commenced, May 1900, in porcelain pots which 1 have already described in the preceeding pages. The general plan of these experiments is given in the following table. 306 M. Nagaoka. UPLAND RICE PLANTS, | N. applied | N. applied Defference Kind of nitrogenous per ha. | per pot. No of pots. of | manures, Kg, | gr. Cultivation. | Tey) 493, 97: Sodium nitrate. oO fe) | 2, 50, 98 > 5 25 0.1227 Without 3, 51, 99 , ” 50 0.2454 irrigation. 4, 52, 100 5 2 100 0.4908 aie | | eee Gp" § SP 10 | Sodium nitrate. fe) xo) | 6, 54, 102. + _ | 25 0.122 With | ' | i; 55, Ao35 a ” 50 0.2454 irrigation. 8, 56, 1o4 ” ” | 100 be) 4908 9, 57, 105. | Ammonium sulphate fe) fe) 10, 58, 106 ” » 25 0.1227 Without II, 59, 107 "9 7 50 0.2454 irrigation. 12, 60, 108. | eS S 100 0.4408 >. | a eee ee | 13, 61, 109 | Ammonium sulphate, fo) fe) 14) (02, 10; - ; 25 0.122 With. | 15, 63, 111. | » 2 50 0.2454 irrigation. 16, 64, 112 | ” » | 100 0.4908 : | Vio) Obs alias Sodium nitrate, | fe) | o j 18, 66, 314, :5 4 | 25 0.1227 | Without 19, 67, 115. | % ” | 50 0.2454 irrigation, 20;, 68, 16, 7 is | 100 0.4905 - ae) hace | | 2%, 69, 117. Sodium nitrate, | oO fe) 22, 79, 118 ” ” 25 0. 1227 With 23, 71, 119 ¢ 33 50 0.2454 irrigation, On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 307 ros 2k Ammonium sulphate. fo) fo} 26, 74, 122. 9 ” 25 O11 227 Without 27, 75, 123. ” i 50 0.2454 irrigation, 28, 76, 124 ” » 100 0.4908 | ZO 77a) 125 Ammonium sulphate. fo) fe) | ge, 7778)" 126. » 2 25 0.1227 | With 31, 79, 127. » 3 50 0.2454 | irrigation. 3255. &o, “128. A c. 100 0.4908 In the begining of May, all the pots received about 2 Kilograms of small pebbles besides 11 Kilograms of the air dry and well sifted soil which had been taken from one of our unmanured experimental dry fields. For the experiments with the paddy rice plants, the soil of dry fields is certainly not favorable but for sake of comparison this had to be taken. The general manures, at the rate of 100 Kilograms of potash as potas- sium sulphate and phosphoric acid as double superphosphate repectively per hectare, were mixed with the weighed soil which was then filled in the pots. The sodium nitrate and ammonium sulphate were, in the form of a solu- tion, given on the 14th of May. On the next day, one half of the pots which were to be cultivated without irrigation, received, 9 healthy seeds of the up- land rice plants (the Variety Terishiradsu) and the other half paddy rice seeds (the Variety Satsuma). On the same day, the remainders of the above two kinds of the rice seeds, were, as a preparation for other pots under irrigation, sown upon a seed bed specially prepared for the purpose and after the germination of the seeds, the young rice plants in the bed were nourished and treated in the usual manner. When the young rice plants were 44 days old, they were transplanted in the designated pots, which had been previously and sufficiently irrigated and stirred. Each pot received 9 healthy equally sized plants in 3 bundles. During the whole vegetation there were observed some differences of growth between the plants with and without irrigation; thus the former 308 M, Nagaoka. plants in all cases gave out less tillers than the latter, and they had rather slender and somewhat narrow leaves while the plants without irrigation possessed, on the contrary, more tillers and their leaves were long and broad. The plants which received the nitric nitrogen exhibited a pale colour but their significance was not so eminent as in the cases of the preceding experiments, they bloomed about 2 days and ripened 5 or 6 days earlier than those plants which were manured with the ammonical nitrogen. The general differences will be seen in the plates of the last pages. On the 2nd of November, all the plants were harvested at once and at that time, the plants without irrigation were just in the state of full ripeness while those with irrigation were in a condition of a little over- ripening. The yields per pot and the averages of the 3 parallel expriments are given in the following 4 tables in which I give also the qnantities of the total nitrogen contained in the whole crop per pot. On the Behavior of the Rice Plant to Nitrates and Ammonium Salts, 399 FABEE. vJ. Results of the upland rice plants without irrigation. a N. ap- = oa Sous Average per pot. N, in the ‘ tei ae Kind of plied No Straw. | Grains, coe NS crop the per ha. of Straw. | Grains. | Total. | per pot. ots, i S ie manures, Ken | nicles, gr ae gr. gt gr gr 1 oO 15 14.9 12.5 49 5 4 17 14.2 | 89 S 97 ES ” 15 15.1 II.o i 14.95 10.55 25.50 0.160 9 2 Fe 16 14.2 12.5 a 75 = 9 14 14.5 10.0 105 Fe 14 16.8 8.4 bi 25 20 21.5 (Sey 2 = § 5 20 23.1 9.0 21.67 8.90 | 30.57 | o.I9g1 a 8 » 15 20.4 9.7 50 17 25.4 14.9 ” ’ ” 15 23-4 14.1 24.70 14.37 39-07 0.228 i 19 25.4 14.1 100 16 25.4 12 ; . her > 22 33-4 12.6 30.63 1X77 42.40 0.265 ” 23 33.1 bre = — —— oo — —~ ee — 5 i 25 17 21.8 13.4 & 3 2 A Py 17 21.4 TRF 22.33 11.37 33.70 0.223 & Ss =e _ 19 23.8 9.0 50 2I 2 14.2 See 35 3 16 26.6 14.5 26.90 14.30 | 41.20 | 0.248 310 M. Nagaoka. TABLE «i: Results of the upland rice plants with irrigation. eS N. ap- : reece |e Average per pot, N, in the No of Kind of plied No Straw. | Grains, total crop the per ha. of Straw. | Grains. | Total. | per pot. pots. manures. Ke. panicles. Z e 5 gr. gr or 5 oO Ise | 132 9.4 53 o » 12 13.8 7-4 (e) 101 = > 12 13.1 9.2 3 F307 9.27.) 22447) On75 13 2 3 12 12.4 10.2 | 61 eS “4 14 13.6 10.2 | 109 » 13 13.0 9.2 | = = ps | 6 s 25 12 16.7 9.0 / s o | 54 - s 5 15 16.8 9.9 16.50 9.83 26.83 | 0.223 | 2 2 | 102 9 16 16.0 10.6 | 7 50 16 211 14.7 55 eth ’ 15 22.0 14.0 21.53 14.10 35.63 0.247 103 cE 13 21.5 13.6 ee | ee | ee } 8 100 21 33.1 22.2 56 Ay as 5 25 36.6 22.5 34.33 21.30 55-53 0.379 104 + 21 428 19.2 14 = F 25 15 17.5 12.0 - 62 : af - 15 16.9 10.7 17.17 | 11.37 | 28.54 | 0.247 110 = a 5 13 Wey 11.4 15 50 19 24.4 15.5 63 » %» 16 23.7 15.1 23.87 | 14.70 | 38.57 | 0.260 111 » 18 23.5 13.5 16 1co 22 40.3 24.0 64 1» oe ” 25 41.3 22.4 41.93 | 23.10 | 65.03 | 0.384 On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. ake | PABLE ¢ Lit. Results of the paddy rice plants without irrigation. x: N, ap- Aa eet foe Average per pot. N. in the No of Kind of plied No Straw. | Grains. : total crop the per ha, of Straw, | Grains. | Total. | per pot. pots. manures. Kg. panicles. gr. or. a gr. er. gr. 17 o 14 20.2 11.8 65 2 99 14 23.1 13.9 113 | 10 13.5 12.7 tH 19.85 12.80 | 32.65 | 0.1995 25 5 ” 15 19.1 They S 73 = » 15 19.1 11.9 12! a 14 20.1 14.8 18 25 15 19.7 I1.7 same 66 3 5 ” 14 21.9 12.7 21.23 12.13 33-36 0.219 a s II4 ay 13 221 12.0 19 5° 19 30.2 13.4 | 67 es 22 18 28.0 14.4 | 30.10 | 14.80 | 44.90 | 0.250 115 9 19 32.1 17.0 20 100 28 38.8 18.7 68 ees » 25 38.1 17.0 | 38.97 57.17 | 0.348 116 0 26 40.1 18.9 26 ef 3 25 20 26.8 15.7 & 3 2 74 2 2 ” 14 25.4 15.9 26.50 41.87 0.269 oF rs 122 a » 16 27-3 15.5 312 M. Nagaoka. PABEES ive. Results of the paddy rice plants with irrigation. © ft N. ap- = eves ieee Average per pot. N.in the RE Kind of plied No Straw. | Grains. = total crop the per ha. of Straw. | Grains. | Total. | per pot, ots. ; ] manures. Kg. panicles. gr. gr. or, gr, or, or, 21 fe) 13 16.0 10.7 68 5 , 12, 18.0 12.0 x 117 3 as 12 16.5 10.2 ie 19.33 12.30 | 31.63 0.2345 29 5 » 12 M7) 1337 Fil = ” 15 22.0 13.2 125 » 12 rainy | 14.0 22 Este 25 13 22.2 rane = o is} ~~ 7° 3 s 9 14 22.4 12.9 22.10 | 13.00 | 35-10 | 0.267 i” s 118 2G 15 ey 13.0 23 50 17 30.2 12.9 71 32) » 18 28.4 13.0 29.23 12.87 | 42.10 | 0,287 119 ” 13 29.1 127 120 ” 24 46.4 12.8 30 Be 25 17 25-5 | 13.7 ES 78 fe. , 14 26.5 15.2 | 26.73 | Igq7 aq) 4g2q) eames 126 a a 4 18 28'2 17.5 On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 313 From the numbers of the above 4 tables, some facts can be learned viz., the upland rice can also be cultivated like the paddy rice, with irrigation, whilst the paddy rice plant is also fitted for the cultivation without irriga- tion, although, both species of rice plants, when grown under their normal condition produce better results. As to the effect of the sodium nitrate and ammonium sulphate, it will be generally seen that the latter salt acted better than the former; but before entering into a detailed discussion in regard to their efficacy, I shall calcu- late from the above results, the actual surplus yield over the pot not supplied with any nitrogen and also the actual extra of the assimilated nitrogen. A. Upland rice plants. WITHOUT IRRIGATION, N. applied | Surplus of the |} Surplus of the . x : 5 No of Kind of the per ha. yield. assimilated N, pots. manures. Kg. or. gr, 2250, 98. Sodium nitrate. 25 10.07 0.031 33 5I, 99 ” ” 50 13.57 0.068 4; 52, 100 a x 100 16.90 0.105 1O;, 585. 106 Ammonium sulphate, 25 8.20 0.063 IT, 59, 107 » 9 50 15.70 0.088 12.) 9.60.) “108 ¥ . 100 29.90 0.125 WITH IRRIGATION, 6; 54}. 102. Sodium nitrate. 25 4.39 0.048 For 1593, 0203 » 39 50 13.19 0.072 8, 56, 104 . 3 100 33.09 0.204 m4.) 62, Tro Ammonium sulphate, 25 6.10 0.072 Moeme OS, LIT 5 3 50 16.13 0.085 314 M. Nagaoka. B. Paddy rice plant. WITHOUT IRRIGATION, N.. applied Surplus of the | Surplus of the Moves Kind of the per ha. assimilated N, pets. manures, Kg, gr. 18, 66, I14. Sodium nitrate, 0.0195 Yoo 167.7315 ~ 0.0505 207 68: ITO = 0.1485 1S R eR EG Ammonium sulphate. 0.0605 27, 75; 123. > » 50 0.1075 28, 76, 124 : é 100 0.1785 WITH IRRIGATION. 2255 JO EIS. Sodium nitrate. 25 3.47 - 0.0325 23, 71, 119. * ” 50 10.47 0.0525 FA 2. 20: fj 100 30.61 0.1685 30; 78, 126. Ammonium sulphate. 25 10.57 0.0825 31, 79, 127. ” » 50 15.17 O.1II5 32, 80, 128. = < 100 32.31 0.2085 Thus it is generally seen from the above calculations that the ammo- nium sulphate acted upon both upland and paddy rice plants better than the nitrate, the latter being always less absorbed than the former; nevertheless it is also observed that the nitrate was more utilized by the upland rice plants then by the paddy rice. Although these special properties ofthe upland rice which enables it to utilize nitrate better than paddy rice does have not yet been ascertained by any physiologist, yet it may be deduced from some observations of Loew! and U. Sudsuki,? that a higher concentration of sugar is attained in the 1 Loew, The energy of living protoplasm, ? Bulletin, College of Agriculture, Imperial University, Vol, III, No. 5. On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 315 leaves of the upland rice plants, whereby the nitrates can be more easily transformed into asparagin and protein. For the sake of better comparison I have calculated here, assuming re- spectively the surplus yield and also the surplus of the assimilated nitrogen caused by the medium dose (50 kg. per ha.) of the ammonium sulphate, to be 100, the percentages of the yield and assimilability for the nitrate applied in the same quantity (50 kg.) as follows :— UPLAND RICE PLANT, Without irrigation. With irrigation. Surplus yield. 86.4. 81.9 of the assimilated N, 77.3 $2.4 PADDY RICE PLANT, Surplus yield, 71.3 | 69.0 » of the assimilated N, 47.0 47.0 Accordingly it is quite clear that in all cases the ammonical nitrogen has displayed a better action than the nitric nitrogen, while it also becomes obvious that the upland rice plants have utilized 30% more nitrogen in the case without irrigation and 35% more with irrigation, from equal dose of the nitrate than the paddy rice plants. Although the paddy rice plants in these experiments have utilized the nitric nitrogen less than the upland rice plants, yet when these results are compared to those of the Series I and II, it will be seen that the paddy plants took up a somewhat higher amount of the nitric nitrogen than in the former cases. This difference of the nitric nitrogen assimilation will most probably be due to the difference of the soil used, since in these experiments dry field soil has been used instead of paddy field soil. Concerning the chemical composition and physical properties of our dry and paddy field soils, there is no great difference, as we have frequently 316 M. Nagaoka. described in our former bulletins,’ yet the contents in humus, protoxide of iron and in alumina are evidently greater in the paddy soil than in the dry field soil. Hence it might be inferred that when a nitrate is applied to the soil rich in humus and protoxide, a part of the salt is lost by denitrification. IV. Experiments on the influence of lime compounds upon the effects of sodium nitrate, ammonium sulphate and fish manure with paddy rice plants. It is well known that lime and its compounds have some beneficial effect upon the absorption of nitrate nitrogen by plant roots; but such observa- tions have been made on the ordinary dry fields. It seemed to me of some interest to observe also the behavior in the paddy fields. My experiments were carried out, in the year 1901, just in the same manner, as in the first series, with 120 porcelain pots in the glass house; the soil was also taken from the same paddy field as in the first experiments. The general design of these experiments is shown in the following table:— ' The Bulletin, College of Agriculture, Imperial University. Vol I, No, g and 11, On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 317 WITHOUT NITROGEN, tiga eee ies [caches |p Sedotine | AQ eae! pots. Kg. gr. compounds, gr. Ie 4is Ole fe) fe) e) oO 2, 42, 82. o : fo) Caustic lime 19.632 a7 43. 83. fo) fe) Calcium carbonate 19.632 Ay AAS. SA. fe) fo) Calcium sulphate 19.632 WITH SODIUM NITRATE. | ome 455 S5< 50 0.2454 o | oO 60) 46; 86: 100 0.4908 o | oO Tee 847. -* S87; 150 0.7362 fe) | fe) OMA OO 50 0.2454 Caustic lime 19.632 Gh 49; So. 109 0.4908 = ” 1054, 50, 90. 150 0.7362 3 = ii Shae ue 50 0.2454 Calcium carbonate - L274) 52, G2: 100 0.4908 “ + 13, 53, 93: 150 0.7362 ' | ” Kew 54; “G4: 50 0.2454 Calcium sulphate | = Toe See OSs 100 0.4908 $3 TO} 50s) 90: 150 0:7362 4 WITH AMMONIUM SULPHATE. Bivaa nds. O¢-) | 50 0.2454 | fe) | oO 15.) +50). OSs 100 0.4908 oO oO 19, 5% 99 | 150 0.7362 | Oo | Oo 20, 60, 100. 50 0.2454 Caustic lime 19.632 21, 61, 10%. | 100 0.4908 = Soe: 102: 150 0.7362 25 O5;, LOZ 50 0.2454 | Calcium sulphate mans O04, 104. 100 0.4908 | “ | » M. Nagaoka. No. of en ene ee ha. Kind of lime CaO pees pots. Kg, or, compounds, er, 2h Obs OG. 150 0.7362 Calcium sulphate 19.632 26,9 (660, 106: 50 0.2454 rs s 2707, 107: 100 0.4908 5 s Zoos, TOS: 150 0.7362 - ‘ WITH FISH MANURE. 29, 69, 109. 50 0.2454 fe) fe) Blot fo, histo» 100 0.4908 fe) fo) Bhi PUAty iGiiic 150 0.7362 fe) fo) a2, 72," 112) 50 0.2454. Caustic lime 19.632 Se ee ish 100 0.4908 A 55 34, 74. 114. 150 0.7362 ” ” Cie oii Nile 50 0.2454 Calcium carbonate " 26,000; Il: 100 0.4908 x os hie Uhre SE 150 0.7362 "5 -: Stor, Gps) DM ey 50 0.2454 Calcium sulphate A 39; 4793, LL: 100 0.4908 ” . 40, 80, 120. 150 0.7362 a 5 On the 21st of June, all the lime compounds were added to the soil at the rate of 4000 Kilograms of CaO per ha which makes 19.632 gram per pot, and as the general manure, phosphoric acid in the form of the sodium phosphate and at the rate of 200 Kilograms per hectar and the potassium sulphate at the rate of 150 Kilograms of K,O were applied on the 23rd while the nitrogenous manures! were applied on the 24th of June. The transplantation of the young rice plants took place on the 25th and each pot received 9 equally nourished plants in 3 bundles. ' The fish manure was prepared from sardine and contained 8,9019% of nitrogen in the air dry state, On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 319 A fortnight after the transplantation, all the plants displayed a quite distinct and different development according to the nature of the nitrogenous manures; thus the plants which had received the ammonium sulphate grew the best and those with the fish manure came next while the plants with sodium nitrate remained pale and small in size. The general difference of the growth will be seen from the plates of the appendix. All the crops were harvested on the 26th of November with the follow- ing results ;1 the numbers of the last column are the quantities of the total nitrogen in the whole crop per pot. 1 The detailed figures per pot are given in the appendix of the last page. WITHOUT NITROGEN. 4 N. applied | Compound Straw, Grains. Total crop, | Assimilated No of per ha. £ nitrogen r per pot. pots Kg lime, gr gr gr gr, eat Or fo) fo) 16.67 12.97 29.64 0.2045 A PGS tel fe) CaO 17.50 15.87 33-37 0.2536 a AS, Sz fo) CaCO, 16.50 14.17 30.67 0.2202 45) 44;. 84 fo) CaSO, 15.17 12.67 27.84 | 0.1836 Bsee4 ds 29.56 0.2055 6, 46, 31.56 0.2231 Wes 30.76 0.2027 8, 48, 41.90 0.3089 9, 49, 42.00 0.3205 10, 50, 38.57 0.2575 Big) 51; 32.77 0.2256 12, 52, 33-63 0.2256 13, 535 33-63 0.2263 é 320 M. Nagaoka. i N. applied | Compound Straw. Grains. | Total crop. | Assimilated No of per ha, P nitrogen > per pot pots Kg lime gr. er. gr gr 14, 54, 94. 50 CaSO, 17.00 13.43 30.43 0.2006 15. 554 OSs 100 ‘ 17.33 13.37 30.70 0.2015 16, 56, 96. 150 os WEL. 13.33 30.50 0.1892 WITH AMMONIUM SULPHATE. 17, 57, 97- 50 o 30.50 22.77 53-27 0.3537 18, 58, 98 100 fe) 42.67 31.67 74.34 0.5062 19, 59, 99 150 fe) 49.67 40.43 90.10 0.6763 20, 60, 100 | 50 CaO 32.00 | 26.07 58.07 0-3856 21,, 6%, 101. | 100 - 35-33 35-13 70.46 0.5982 | 22, 62, 102 | 150 a 39.33 | 39:23 78.56 0.6797 23 63, 30g 50 CaCO, 29.50 | 24.87 54-37 | 0.3812 7 24, 64, 104 100 9 37.83 30.83 68.66 0.4834 25, 65, 105 150 ~ 44.33 37-83 $2.16 0.6272 26, 66, 106 50 CaSO, 28.67 21.33 50:co 0.3281 27, 67, 107 100 ¥» 7 Sy ee 74.27 0.5587 28, 68, 108 150 55 47.67 39-17 86.84 0.7078 WITH FISH MANURE, 29, 69, 109. | 50 o 27-33 20.50 47.34 0.3154 30, 70, 110 100 oO 33.67 209.40 63.07 0.4224 3%, 71, 111 | 150 ) 47.50 37-43 74.93 0.4835 32, 72, 112 | 50 CaO 28.67 25.13 53.80 0.3865 33) 73, 113 100 ” 38.50 31.87 70.37 0.4902 On the Behavior of the Rice Plant to Nitrates and Ammonium Salts, 321 Bist 5.115. 50 | Ca CO, 25.67 20.23 43-90 0.3111 26, 70, 116. 100 i" 36.67 30.67 67.34 | 0.4624 Siw DALY E 150 » 44.67 36.90 | 81.57. | 0.6439 35, Jox118. 50 | CaSO, 24.33 18.17 | 42.50 | 0.2767 39, 79, 119. 100 | - 32.67 25.40 58.07 | 0.3795 40, 80, 120. 150 | a 41.00 34.27 75.27 | 0.5235 In considering these results, it will be observed that the ammonium sulphate had again a considerable good influence upon the production and next to it also the fish manure. Before entering into a farther discussion of these results, I have calcu- ‘lated in the following table the extra yield caused by the 3 nitrogenous manures over the no nitrogen pot and also the extra of the assimilated nitrogen. WITH SODIUM NITRATE. N itrogen applied Gampound Extra, No. of pots, per ba. kg. | ee Yield. | Assimilated nitrogen. gr. gt. 5, 45, 85. 50 | fo) (—) 0.08 0.0010 6; (26, 86, 100 oO 1.92 0.0186 7 47, 87. 150 oO I.12 | (—) o.oo18 8, 48, 88. 50 CaO 8.53 0.0553 9, 49, 89. 100 ‘ | 8.63 0.0669 BO, %50;.0 90. 150 ‘ 5.20 0,0039 Mie hs? OF. 50 CaCO, 2.10 0.0054 12, 52, 92. 100 . 2.96 0.0054 13) .53: 93> 150 . 2.96 | 0.0061 145 (54 94: 50 Caso, 2.59 | 0.0170 15, 55» 95. 100 7 2.86 0.0179 16, 656; » 96. 150 $5 2.66 | 0.0056 322 M. Nagaoka. WITH AMMONIUM SULPHATE. | Nitrogen applied Compound Extra. No, of pots. per ba. ao kg. of lime. Yield. Assimilated nitrogen. gr. gr. ie ype COVE 50 fe) 24.23 0.1512 18, 58, 98 100 fe) 44.70 0.3017 19, 59, 90: 150 fe) 60.49 0.4718 20, 60, I00, 50 CaO 24.63 0.1320 215 OI; 101: 100 °F 37.09 0.3446 225 | O2; = 312. 150 5 45.19 0.4261 25,2 Os: 50 CaCO, 23-70 0.1610 24, 64, 104. 100 . 37.99 0.2632 PI SIS itSis 150 = 51.49 0.4070 26, 66, 106. 50 CaSO, 22.16 0.1445 2s OTe ator 100 - 46.43 0.3751 28, 68, 108. 150 5 59.00 0.5242 WITH FISH MANURE. On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 323 These calculations show that the sodium nitrate, in spite of its large dose, had no great effect while the other two manures (ammonium sulphate and fish manure) gave a very satisfactory crop in proportion to their doses. These results, in general, coincide well with those of the series I and IJ and hence it may be justified to conclude that paddy rice plants when cultivated in a genuine paddy soil like our’s, have no sufficient capacity to assimilate the nitric nitrogen. Hence the nitrogenous manures to paddy fields should be applied either in the form of ammonium salts or of organic manures. In regard to the beneficial influence of the lime and its compounds upon the absorption of the nitrate, it becomes clear from the above figures, that all the lime compounds have accelerated the assimilation of the nitric nitrogen to a certain extent. The caustic lime was most efficient whilst the calcium carbonate and sulphate remained but little behind. Though the action of the caustic lime upon the nitric nitrogen assimi- lation was quite evident, the total increase of the organic matter was not sufficient enough to recommend the sodium nitrate as a profitable manure for paddy rice. As to the ammonium sulphate, the lime compounds had no favorable but, on the contrary, an unfavorable effect on the assimilation of the nitrogen ; only in one case (medium dose (100 kg.) of the ammonium sulphate with calcium sulphate) was an exception. The considerable diminution of the yield caused by the caustic lime in the pots with the ammonium sulphate, is doubtless due to the high alkalinity of the lime by which the ammonia of the sulphate was set free, causing a loss of active nitrogen. As to fish manure the caustic lime seems to have acted somewhat benficially but it was not so noticeable as in the case of the sodium nitrate. This action exerted by the caustic lime upon the fish manure is undoubtedly due to the influence upon the decomposition of organic compounds. 324 M. Nagaoka. VIEEDS PER POT. No. of pots. Straw. Grains. No. of pots. Straw. | Grains, gr. gr. gr. | gr. I 16.0 11.5 9 23.0 19.0 41 17.0 12.7 49 22.5 19.5 81 17.0 r2a7 89 22:5 19.5 2 17.0 15.8 10 17.0 13.0 42 17.0 15.8 50 23.0 19.7 82 18.5 16.0 go 23.0 20.7 3 17.0 15.0 II 18.0 14.3 43 16.5 14.0 51 18.0 16.5 $3 16.0 13.5 gl 17.0 14.5 4 15.0 nDS | 12 19.0 15.3 44 15.0 12.0 52 18.5 15.5 84 15.5 13.7 92 1755 15.1 5 16.0 1317 13 17.0 13.2 45 16.0 13.5 53 19.0 16.2 85 17.0 12.5 93 19.0 16.5 6 17.0 14.7 14 17.0 14.0 46 17.0 13:5 54 17.0 13.8 $6 18,0 14.5 04 17.0 12.5 7 17.0 135 15 17-5 13-3 47 18.0 14.0 55 17.0 13.8 17 17.0 12.8 95 17.5 13.0 8 22.0 20.6 16 17-5 13.5 48 22.0 18.5 56 17.0 13.0 Sa On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 325 YIELDS PER POT. No, of pots. Straw. Grains. | No. of pots. Straw. | Grains, gr, gr. | er. gr, 17 31-5 23-1 | 25 44.0 37-5 57 30.0 21.6 | 65 44.0 38.2 97 30.0 23.6 105 45.0 37.8 18 43.0 30.0 26 28.0 | 21.5 58 42.0 33-5 66 28.5 21.5 98 43.0 Pitas 106 29.5 | 21.0 19 59.0 39-3 27 42.5 35:9 59 50.0 41.0 I 67 41.0 31.3 99 49.0 41.0 107 40.0 33-0 20 32.0 26.5 | 28 47.0 38.5 60 32.0 26.5 | 68 46.0 41.0 100 B20 Prd) 108 50.0 38.0 i 21 35.0 32.8 29 26.0 20.5 61 41.0 40.3 69 29.0 | 20.8 101 30.0 aR 109 27.0 | 20.2 22 39.0 38.8 30 32.0 20.8 62 34.0 36.3 7o 37.0 30.8 102 45.0 42.6 +8 fe) 32.0 27.6 23 27.5 | 22.0 | 3I 49.0 39:3 63 28.0 22.3 | 71 47.0 37-5 103 33.0 30.3 | III 46.5 36.5 24 38.0 31.0 32 28.0 25.7 64 38.0 30.5 y 20.0 24.2 104 37-5 31.0 112 29.0 25.5 326 M. Nagaoka. VIBE DS PER LOW i No. of pots. Straw, Grains, || No. of pots. Straw, Grains, gt gr er, gr eee ed | eee, a 33 38.5 32.5 ! 37 45.0 367 || 73 38.5 33.0 77 44.0 37.0 113 38.5 30.1 | ryiy 45.0 37.0 34 45.0 38.8 38 24.0 18.0 i 74 38.0 36.5 1 78 25.0 18.5 114 44.0 41.2 l 118 24.0 18.0 35 27.0 22.2 39 33.0 24.2 75 190 16.5 79 33.0 25.5 15 28.0 22.0 | 119 32.0 26.5 36 36.0 30.8 ! 40 40.0 33-5 76 38.0 30.0 } So 41.0 35.8 116 36.0 31.2 ) 120 42.0 | KE V. Experiments on the action of a sodium nitrate and ammonium sulphate upon other aquatic agricultural plants. Since there are, in Japan, several agricultural plants which are general- ly cultivated under the same conditions as paddy rice plants, it will not be uninteresting to investigate also the effects of sodium nitrate and ammonium sulphate upon some of such plants: and as the experiments of this sort have not get been tried, as far as I know, among our agricultural chemists, the results of these investigation will be of some value in a theoretical and practical view. A. Experiments with Funcus effusus L. These experiments were tried, in 1900, and in conjunction with the series III. The soil also came from the same dry land. The treatment ——— bo NI On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 3 of the Juncus were also similar to those of the rice plants in the 3rd series of experiments. The rate and actual quantity of the nitrogen applied to each pot are as follows. Kind of Nitrogen applies Nitrogen applied No, of pots, nitrogenous per ha. per pot. manure, kg. gr. 33, 81, 129. fo) fo) fo) 34, 82, 130. Am, sulphate, 25 0.122 The crops were harvested on the 2nd of November, with the following results. WITHOUT NITROGEN. Nitrogen Yield Assimilated | Surplus. applied per pot, Average nitrogen No: af pots. per ha, per pot. Assimilated kg, er, gr. er. Yield. nitrogen. gr. er. 33 fe) 19.7 | 81 ” 19.7 | 129 9 19.0 20.50 0.1740 37 e 21.5 | 85 aS 21.5 | | 133 eS 21.6 328 M. Nagaoka. WITH SODIUM NITRATE. Nitrogen Yield Assimilated Surplus, No. of pots. re aa ace ee Assimilated kg, gr. gr. er. Yield, nitrogen. J) —E——————— 34 25 19.4 82 | % Zon 22.37) 0.2082 1.87 0.0342 130 5 25.6 35 59 24.7 83 ip 25.2 23.83 0.2225 3:33 0.0485 131 5 24.6° 36 100 31.3 84 a 28.5 29.03 0.2531 8.53 0.0791 132 | 5 27.3 Thus it will be seen that the Fuzcus effusus utilised, like the paddy rice plants, more nitrogen from the ammonium sulphate than from the sodium nitrate, that hence the ammonium sulphate should also here be preferred as a manure to the nitrate. On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 329 As to the effect of the different doses of the nitrogen, it is seen that the larger the quantity of both salts was applied the larger was also the yield. Since these experiments were, in contrary to the common practice, ' carried out with a dry field soil, I took up again, in 1901, a second series with ordinary paddy field soil. The soil was same as that of the IV series of experiments ; the methods also were just the same as in the preceding experiments. The crops were harvested on the 26th of November with the following results. WITHOUT NITROGEN. Nitrogen Yield Assimilated Surplus. = applied per pot, Average. nitrogen —- No. of pots, per ha, | ig per pot. Assimilated kg, gr, | gr. er, Yield. nitrogen, er, er, I oO 16.0 21 ; 15.0 41 ” 13.0 14.50 0.1720 6 9 14.5 24 5: 14.0 46 : 14.5 | 2 25 21.0 | | 22 ks 16.0 18.67 9.1874 4.17 | 0.0154 42 9 19.0 3 50 19.0 23 9 19.0 | 17.83 0.2047 | 333 0.0327 43 ’ 15.5 | | 4 100 17.5 24 ” 19.0 17.50 0.1795 | 3.00 | 0.0075 44 : 16.0 | 330 M. Nagaoka. Nitrogen Yield Assimilated Surplus, No. of pots. es an sags rene moc Assimilated — kg, er, or, er, Yield. nitrogen. S| eee 5 150 15-5 25 Pe 15.0 15.33 0.1610 0.83 (—) o.o110 As , 15-5 WITH AMMONIUM SULPHATE | 25 Bear 27 “3 Ase 22507 0.2379 4) 7.67 0.0659 47 21.5 8 50 22.5 28 4 22.0 22.83 0.2669 8.33 0.0949 45 + 24.0 9 100 31.0 29 = 29.0 | 30.00 0.3924 15.50 0.2204 49 30.0 | 10 150 2210 30 | ¥ 31.0 | 31.67 0.5231 17.17 0.3511 50 , 32.0 Accordingly the ammonium sulphate had again a better manuring action than the nitrate and the difference of the influence of both salts are, in general, still wider than in the former experiments. Besides, the peculiar fact is observed in the above results that the yields of the nitrate pots gradually decreased with the increase of the nitrate in the soil whilst in the pots of ammonium sulphate the yield was gradually increased proportional to the quantity of the nitrogen applied. Denitrification or formation of the poisonous nitrites from nitrate may account for this. At last, when the actual surplus of the yield and of the assimilated On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 331 nitrogen, caused by the 50 kilograms of the ammonium sulphate are assumed respectively to be 100, the ratio of the increase caused by the same dose of the sodium nitrate will be 40 for the plus yield and 34.6 for the surplus of the assimilated nitrogen, therefore the relative value of the nitrate 40+ 36.0 2 for the Juncus plants will be = 37.3 for 100 of the ammonium sulphate. B. Experiments with Arrow-heads, (Sagittaria sagittifolia L.) These experiments were performed under the same conditions and treatment as the preceding experiments A. (1901). The pots were 30 in number. Each pot received one healthy tuber of arrow-head. On the oth of September, the roots and upper parts of the plants were harvested separately. At this time, the new tubers in the roots were very small but some of the leaves begun already to die. The crops, after having been completely air dried, were weighed and analysed with the following results. WITHOUT NITROGEN, Noloft Nitrogen Crop per pot. Average per pot. N. in the Surplus. - applied total crop | per ha. | Upper Upper per pot. | j |Assimilated Ee kg. part. Roots, | part. Roots. | Total. er. Yield. | nitrogen. er. er. er. er. or. er. II fo} 18.8 19.0 31 ” 18.0 21.6 51 : 19.6 20.5 18.22 | 19.50 | 38.12 0.39¢6 16 » 17-3 17.0 36 » 20.0 20.5 332 M. Nagaoka. WITH SODIUM NITRATE, YS. GE Nitrogen Crop per pot. Average per pot. N. in the Surplus. pone applied total crop t per ha. | Upper Upper per pot. Assimilated aie: kg part Roots part er. Yield. | nitrogen. 4 gr. on gt. gr. i a On the Behavior of the Rice Plant to Nitrates and Ammonium Salts. 333 The arrow-heads thus took up a considerably larger quantity of the nitrogen from the ammonium sulphate and their yields were also uncom- paratively greater than when supplied with the sodium nitrate. With the largest dose (150 kg. per ha.) of the sodium nitrate a decrease of the crop was also observed, hence it may be assumed here that denitrifi- cation had, as in the case of Fuucus, taken place. By qualitative tests, the presence of nitric acid, was, as in the case of the rice plants, observed, in both, Juncus and Arrow-head supplied with large quantity of the sodium nitrate, wherefore we may be permitted to believe that these plants have like the paddy rice plants, not sufficient capacity for the transformation of nitric nitrogen into proteids. Now calculating on the same base as in the calculation of Juncus, the relative value is obtained for the sodium nitrate for arrow-head as seinen © + 37-0 33.9 for 100 of the ammonium sulphate. SUMMARY. It was sufficiently proved in all of the preceding trials, that paddy plants cannot utilize nitric nitrogen as well as ammoniacal nitrogen. The causes of this phenomenon may be: I. Paddy plants do not accumulate a sufficient quantity of sugar in the leaves to convert all of the nitric acid, absorbed into protein. The pale yellowish colour of the rice plants supplied with nitrate is probably due to the physiological influence of accumulated nitrate. 2. In paddy soils, denitrification and also formation of poisonous nitrites may take place. Indeed, the pots of land IV of the series of experiments with heavy doses of the nitrate gave a slight Griess reaction for nitrite. In order to test my suppositions as to the denitrification a quantity of paddy soil with some sodium nitrate was kept in flasks well filled. Some nitrogen was indeed gradually developed. Further detailed studies will be made later on. As to the relative value of the nitric and ammoniacal nitrogen upon the 334 M. Nagaoka. paddy rice plant, Juncus and Arrow-head, it is seen that for 100 of the ammoniacal nitrogen, the nitric nitrogen had the following value : With Paddy rice 40 (The result of the 2nd series of experiments.) With Juncus 37 ' With Arrow-head 33 If the relative value for the paddy rice plant (40) in assumed to be 100, the value of the nitric nitrogen will be 90 for the Fuxcus and 80 for the Arrow-head. PEATE XX: BUL, AGRIC. COLL. VOL, VI, Upland Rice plants with Sodium Nitrate. 50 25 50 25 With irrigation. Without irrigation. Upland Rice plants with Ammonium Sulphate. HOO With irrigation. 1) >| 5 2 Without irrigation. BUL, AGRIC, COLL. VOL, VI. PILATE XX Paddy Rice plants with Sodium Nitrate. N. 20 N Z/ N24 N per o 25 50 100 Oo 25 50 100 ha, Kg. $$$" ————————————————— Without irrigation. With irrigation. Paddy Rice plants with Ammonium Sulphate. Hit AY SASS i itet : sale ‘ N LY N per, oO 25 50 100 fe) 25 50 cee) ha, Kg. SS SS ee : Without irrigation. With irrigation. ee oe dig ‘Vane ee Auge pas ane LAr if MPS She « hae eg eS Caria Pe PRs \ ' . ore ace: v mi 2 Ss Sodium Nitrate Arrow-head with ere 4 oe) ES Ao ne — —— On Different Degrees of Availability of Plant Nutrients. BY, O. Loew and K. Aso. The degree of availability of plant nutrients in the soil depends upon the size of the particles, the degree of solubility in water, the readiness of being dissolved by the humic acids of the soil or by the acidity of the root- lets and the extent of the root system. The question of availability has recently been treated especially by Fraps', who distinguishes four factors of availability : Chemically available plant food is that present in forms that can be taken up directly by plants. Physical availability refers to enclosure in soil particles or protection of chemically available food. Physiological availability refers to the difference in power of plants to assimilate food. Weathering availability refers to the conversion of plant food into chemically available forms during the growing season of the plant. We treat here only of some effects of different degrees of chemical avatlability. Effects of different availability of lime and magnesia. It has been pointed out by one of us some years ago that the most favorable ratio for plant growth of lime to magnesia in the soil is altered when the availability of these bases is not equal. The most favorable ratio, 3 CaO : = ee or the lime factor, Mgo Was determined by us? for the condition that the 1 Factors of Availability of Plant Food; Amer. Chem, Journ, vol. 72, No. 1. [1904]. 2 Cf. the contributions of 4se, Furuta, Katayama and Daikuhara in these Bulletins. 336 0. Loew and K. Aso. availability is equal. Then the best ratio was found for cereals=1 or only a little higher while for crops with relatively more abundant foliage =2 or 3 ; for tobacco it is=4, as was recently ascertained by Dazkuhara. An equal availability can e.g. be assumed to exist in the soil if both these bases are present as carbonates or as hydrous silicates or as humates. In cases, however, in which the lime is present as carbonate while the magnesia as hydrous silicate or humate, perhaps a very rare case, the availability is not equal any longer, the ratio of CaO: MgO which enters into the plant body may differ somewhat from the former case, and the physiological effect of the different ratio in the plant will finally be recogniz- ed by the difference in yield—-other things being equal. It is a well known fact, further, that liming with siaked lime is more efficacious than with pulverized limestone, which is due to the most part to the finer condition of the particles of the resulting carbonate, when dissolved lime is exposed to the air.! It is impossible to pulverize limestone to such a degree of fineness Hence also the degree of availability of both these products differs. Still more differs that of burnt magnesia and that of the commercial basic magnesium carbonate from that of pulverized magnesite. A smaller amount of the more easily available form will not only suffice to produce the beneficial effect of the less available form, but also by an undue increase sooner reach the limit beyond which harmful effects will set in. Burnt magnesia is more injurious in overdoses than magnesite in the equivalent amount. Since exact quantitative data as to the respective physiological cffects of the different forms are not yet known, we have re- commended pulverized limestone and magnesite for the correction of very unfavorable ratios of CaO: MgO in soils, since in this way the natural con- ditions of soils was much better preserved than in the application of burnt lime or burnt magnesia. But it is clear that sometimes very large quantities of those carbonates would be required what renders it desirable to apply smaller quantities of 1 It occurs, however, that by insufficient attention the slaked lime left in heaps on the fields turns into a hard rock of carbonate. On Different Degrees of Availability of Plant Nutrients. 337 the more efficacious compounds.! Quantitative studies, however, have to be made on an extensive scale in order to determine how much slaked lime, or artificially precipitated carbonate of lime will produce the same result as a given quantity of finely pulverized limestone and, in the case of a deficiency of magnesia, how much burnt magnesia, or artificial magnesium carbonate (basic), or magnesium sulphate would lead to the same increase in harvest as a given quantity of pulverized magnesite. We propose to call the figures obtained in comparison with 100 parts of the finest pulverized natural carbonates the agronomical equivalents. Vhese figures depend only to a small extent on the chemical equivalent, they are determined mainly by the size of the particles,® solubility in water and acids of the rootlets and humus, and by the nature of the soil. A stiff clay soil will yield different figures from a loose sandy soil, especially when the presence of a considerable amount of hydrous silicates (calcium-zeolithes) in the former can change the nature of the compound applied, as, c.g., of magnesium sulphate. Recent experiments at the Imperial Japanese Central Experiment Station at Meshigahara, near Tokyo, which will be continued, have shown that when lime is present as carbonate and magnesia is applied as sa/phate the best ratio of CaO: MgO which is=r with cereals (or nearly so) in case of egual salts changes into 30: 1 with rice in sandculture, while with a soil rich in calcium zeolithe and relatively poor in magnesia it changed to 7:1.% Thus we have for cereals the following best ratios under different conditions : 1 Also the price must be considered by the practical farmer. Burnt magnesia costs double as much as the same amount of magnesia in the sulphate and about 1 time as much as in the artificial (basic) carbonate. 100 Kilo crystallized magnesium sulphate cost about 7 yen ($ 3,50). 2 Thus, the artificially prepared (basic) magnesium carbonate (¢ MgCO, + Mg (OI1), +4 11,0) being of a high degree of fineness of the particles is of much greater efficacy than the pulverized magnesite, while between artificial and natural calcium carbonate the difference in agronomical effects will be much smaller, 3 These experiments of Aosa?, Datkuhara and Nakamura will be published in the Bulletins of Nishigahara, 338 0. Loew and K, 4s». CaO™ 2 it CaO as CaCO, _ 30 CaO as Ca—Zeolith _ 7 MgO 1° MgO as MgSO, 11 | MgO as sulphate ~~ when both bases are equally | with sandculture with MgO in | with a clay soil from Kuma- available. highly available form. | moto. A recent author has declared that ‘magnesia as sulphate has only a very insignificant effect compared with the carbonate or citrate of magnesia” but this is an erroneous conclusion since he had compared egwal quantities of these compounds and thus reached the point at which magnesia in the highly available form of the sulphate could exert already an injurious effect. If that author would have applied mach /ess of the sulphate than of the carbonate he would soon have observed that the magnesia in that form can produce the same beneficial effect. The leading principle in the application of different magnesia manure must be to procure in equal times equal quantities of magnesia for the plant body and this end can be reached with much smaller quantities of the sulphate than of the carbonate. Beyond the most favorable ratio of CaO: MgO entering into the plant body, soon the line will be reached where an excess of magnesia will depress the yield again, and this line is sooner reached with the sulphate than with the artificial carbonate and sooner with this again than with pulverized magnesite, as was already pointed out above. The great efficacy of magnesia in the form of sulphate had been also long ago recognized by Nodbbe and by Hellriegel. Nobbe applied for 1 Liter of sand in his experiments 1.2 g. tertiary calcium phosphate and only 0.1 g. magnesium sulphate, and Hellriegel for 4 Kilo of sand 4 g. calcium carbonate and 0.18 g. magnesium sulphate for the growth of barley in sandculture. Magnesium sulphate at the rate of 112 Kilo per ha was frequently applied also with good results at Rothamsted; in this case the dose of the simultaneously applied superphosphate was such as furnished as much soluble lime, as corresponded about to the dose of the magnesia applied as sulphate. } 1 Cf, also Larbaletrier, Ann, Agr, 1896. ny? o>) Uo \O On Different Degrees of Availability of Plant Nutrients. The application of magnesia as magnesium chlorid cannot be recom- mended, (except in small doses in special cases) since this compound easily dissociates into base and acid and thus acts very injuriously. In the ap- plication of magnesium sulphate it must not be lost sight of that a slow and gradual action on the lime salts of the soil may take place whereby gypsum results. Our cxperiments showed that on addition of 1 g. calcium carbonate to 500 cc. of a 1% solution of magnesium sulphate 0.015 g. calcium sulphate was formed after two weeks standing with frequent shaking. When tricalcium phosphate was treated in the same manner 0.045 g¢. calcium sulphate was formed within two weeks. Availability of lime in the form of gypsum. It is an old observation that although calcium sulphate exerts in special cases a very favorable effect, it cannot replace slaked lime or calcium carbonate. On soils exceedingly poor in sulphates—and such soils occur more frequently than often assumed—it is the cheapest source of sulphur that can be applicd. Also soils very poor in lime, especially loamy soils (but not dry sandy soils) are benefitted by gypsum to which also an indirect action, the liberation of potassa from certain compounds is ascribed. It has also been observed that many leguminous plants and potatoes are more benefitted by gypsum (200-500 Kilo per ha) than cereals are. Often it is recommended to apply gypsum together with woodash. But here it is not the gypsum which comes into action in the soil, since woodash contains potassium carbonate which transforms the gypsum into calczwm carbonate. The reason why lime in the form of the sulphate cannot replace the lime in the form of the carbonate! is due chiefly to the different degree of availability. It is true, gypsum is more soluble in water? than calcium carbonate,—since 100 p. of water dissolve at 12° C.=0.233 p. gypsum and at 60°=0.251 p., while the carbonate only very sparingly in absence of free carbonic acid,—but the reverse takes place in regard to dilute acids. An aqueous solution of carbonic acid will not dissolve more gypsum than plain 1 Also slaked lime passes of course into this form in the soil. 2 Certain salts, as ammonium sulphate or sodium chlorid increase the solubility a little. 340 0. Loew and K. Ase. water does (Davy). Strong mineral acids augment the solubility of gypsuni somewhat, but dilute organic acids exert only a very insignificant effect. Dilute acetic acid of 1 % will dissolve only traces of gypsum more than plain water. 10g. powdered crystallized calcium sulphate was left in 100 ce. ofa 1 % acetic acid for 2 days at 18-20° C. shaking the mixture repeatedly. The solution was found to contain 0.247 % calcium sulphate, while in the controlflask with distilled water=o 244 %. Hence it may be inferred that also the acidity of the roots does not enhance the absorption of lime in this form, nor has the acidity of the humus any effect on the availability of gypsum. Hence gypsum can in moderate doses neither exert the same beneficial effect as the carbonate nor in excessive doses the injurious effects in that degree as the carbonate or slaked lime. From these facts it could be deduced that the lime content of the leaves would not increase even after addition of a great excess of gypsum to the soil. We have grown barley on a soil to which we added the enormous doses of 5% and 20 % gypsum respectively. The check pots contained the original loamy humus soil and a mixture of this soil with the high dose of 5 % calcium carbonate. As general manure served per pot of 8 Kilo soil: ¢. double superphosphate -~ a ¢, potassium sulphate Lo a3 @. ammonium nitrate. < uw The barley grains were sown (20 per pot) on March 9 and the young plants reduced to 9 per pot, leaving only such as were of equal size, on March 31. On April 17 the average height was: CHECK Wiagarer on cite ticneaicere Cols ee 34.3 cm. CaCO, Seon vercc toe neslee scot 25:0" 33 ASO) Oe ones vel ewacu come Ray diy CaSO] BO sires: VII. f 4: 116.86 = -- 1.33 — : ~ (1) IX e — -— 158.93 — 1.33 - ; 0 he U x. | a — — = - 1.33 206.69 On April 23, seeds of upland rice were sown, 25 in each pot, and the young shoots reduced to 11 on June 6. At the middle of June a great difference in the development of plants became noticeable. Length measurement was made on July 12 with the following result :— 1 Lime was here added in the form of gypsum to produce this ratio 3 : 1. 350 S. Suzuki. Average length | Average length No. of Pots. CaO : MgO. of stalks, No, of Pots. CaO : MgO. of stalks. | | c.m. | | GAD; | it 4221 92.7 Vi. | TER EAT 84.0 II 142 P1 | $7.2 | VII. eae 753 | \ : Ill rigezt et il 87.0 VIII rene ee 61.0 | } IV. 1.40: 1 | IOI 5 | IX O13 or 97.6 V. Ree a 97.0 Ne | OR ey 48.0 | On July 20, cars appeared at first in pot IV. and then followed pots II., III., V., VI. and IX. on July 22. Irrigation was stopped on September 15, as at this time the plants became yellow-ripe. The plants were cut on October 10 and left to dry in each pot. The weight of the air-dry harvest was as follows :— aasags ae ; Relative harvest, Total Weight | Number | Average taking the yield fa . . “ weight of of length | . Addition of phosphates and Séedae listens of in the Check pot pers 7 iow ‘ (VII.) as too. ? Elen on yearing | stalks. magnesia or lime salts, eas g r a c.m Total Seeds 2 S- > |sharvest: ; I. (Bone dust + CaCQ,). 93.7 39.7 17 74.5 184.8 | 193.7 II. (Bone dust + CaSO,). | $3.2 36.2 14 80.6 164.1 176.6 IIT. (Bone dust -+ Ca(NO,),). 102.0 39.4 23 71.3 203.2 192.2 IV. (Na,HPO, +CaCO,). 109.4 51.0 22 67.3 215.8 | 248.8 V. (Bone dust). 95.0 40 5 18 Hee 187.4 197.6 VI, (NasHPO,). 84.0 39.5 17 75.6 165.7 192.7 WAVER Gout) 50.7 20.5 If $2.8 100,0 100.0 VIIL (Na, HPO,) + Large amount of CaCO,. 43.0 | 17.4 10 65.5 84.8 84.9 | IX, (Na, HPO, + Large amount of CaSO,), | 102.0 | 68.5 20 73.8 201.2 334.1 X. (Powdered magnesite + Na,HPO,), 29.2 8.0 15 49.8 57.6 39.0 rom this table we learn: 1. That an excessive liming with CaCO, depresses the yield very much, notwithstanding the phosphoric acid being present in the easily available form of secondary sodium phosphate (See Pot VIII). On the Injurious Effect of an Excess of Lime Applied to the Soil. 3 ui — 2. On the other hand, the application of CaSO, in equivalent quan- tities, under otherwise, the same conditions, led in contrary to the greatest production of seeds. This shows that in VIII not the depression of avazlability of the phosphoric acid was caused by the liming, but a depression of the assimilability of the phosphoric acid in the cells themselves. The great difference of the action between the calcium carbonate and calcium sulphate is very easily explained by the fact that the CaSO, is only absorbed from the soil in the measure as it is soluble in water, which is but little, while the absorption of lime in form of CaCO, depends chiefly on the aczdity of the rootlets, and hence the amount of lime which is taken in this form into the plant body is much larger than the amount of CaSO,.! 3. That the powdered magnesite added in such a ratio that the amount of magnesia became three times as high than that of lime led to a very great depression in the yield. 4. Comparing VI. with V., the seed production was almost equal, showing that the action of phosphoric acid in the form of bone dust and of di-sodium phosphate was nearly the same. 5. The application of a moderate amount of lime together with the bone dust has not noticeably diminished the yield (compare I with V) which may be explained by the fact that the soil applied contained 119 of humus, and since the humus has more or less an acid character it would be explained why the CaCO, did here in the small quantities (applied 12 grams per pot) not depress the availability of bone dust phosphoric acid, while the increase from 12 to 116.8 grams. depressed the yield more than 50% in seeds. 1 Only small quantities of gypsum could have been transformed into phosphate, since the amount of sodium phosphate relatively to gypsum was but small. Is the Availability of Phosphoric Acid in Bone Dust Modified by the Presence of Gypsum ? T. Katayama. The investigations of Kel/ner and Léttcher of Nagaoka, b. Schulze and Séderbaum have shown that the availability of phosphoric acid in bone dust is very much depressed by the presence of calcium carbonate while that availability in the secondary and primary calcium phosphate is not essentially altered. The question can now arise, how has a soil, very poor in lime, to be provided with lime, if there is no other phosphatic manure at hand but bone dust? The idea suggested itself that here the application of gypsum in place of carbonate of lime or slaked lime would be in order. This led me to compare the CaCO, and the calcium sulphate in this regard. The sand serving for this culture was treated first with conc. hydrochloric acid and washed with distilled water until every trace of HCl was removed. Each pot contained 2.5 kilo of this sand and was manured with 1.5 gram bone dust (0.303 grams P,O,;). The pots received further the following ratios of powdered limestone, powdered magnesite and gypsum, i CaCO, =3.6 grams. MeCO, =4.18 |,, ll CaSO ,2aq =6.19 grams. MgCO,=418 CaCO, =3.6 grams. ee MeCO Tan |; IV. CaSO, 2aq =6.19 grams. MeCO. =F.04 ? 354 T. Katayama. For further manure a solution of 0.6 NaNO, 0:6 (NERO), 0.4 K, SQ, 6.1 Pesos was prepared and of this solution 50 c.c. were first applied at the starting, bin) LOO7C.c- bt O) on June 3, while another dose of 33 c.c. was applied, on July.21. Upland rice served for this experiment of which 7 young shoots (2-2.5 c.m. high) were transplanted into each pot from the seed bed. The irrigation was carried out upon the principle that 65% of absolute water capacity was almost continuously present. As early, as on the 18th of June, very remarkable differences were observed, the gypsum plants being much more luxuriantly developed than the carbonate plants and showing further a nice dark green color, while the carbonate plants had a yellowish appearance. The average height was as follows: July 1. July 2r. Ts 15 c.m. 30 C.m. II 30° ay Se) eb lt 14 >, 55) a IV. 32 » 5 iets: While furthermore the ears with the gypsum plants No. II and 1V had developed on August 16, there was no ear developed even 4 weeks later with the carbonate plants I and IJ. The plants were cut Sept. 15, and weighed in the air dry state with the following results : 1 See/horst had observed that bone dust can be better utilized when nitrogen is applied as am- monium sulphate than when applied as sodium nitrate, but in our case the calcium carbonate interfered, Average height. Weight of grains. Total weight. Gn: | grams, grams. : : | : 30.0 0.00 3.2 II 67.0 6.50 21.5 Ill 31.0 0.00 4.8 IV. 69.8 7.9} 28.0 This result shows 1) that the availability of P,O; in bone dust was not prevented by gypsum what could already foreseen by the theory, 2) that an increase of magnesia as magnesite did not act favorably. Since the availability of gypsum depends not upon the acidity of the rootlets! but simply upon the rather low solubility in water, an excess of gypsum will under most conditions not act so unfavorably as an undue corresponding excess of carbonate of lime. One more experiment was made in which phosphoric acid was applied as secondary calcium phosphate,? all other conditions were the same as in II and IV mentioned in the above experiment. Also the time of starting and watering of the upland rice were the same. The final result was as follows : Average height. Weight of grains. Toial weight. c.m. grams. grams, Ti: 69.0 FAS 27.0 IV. 71.0 7.63 28.0 Also from this result it will be seen that a certain excess of lime in the form of gypsum over the magnesia as carbonate in a soil does not depress essentially the harvest of rice. Finally it may be pointed out that while in my experiments in which humus was absent the depression of the availability of bone dust by calcium 1 Compare the article of Zoew and Aso in this Bulletin, 2 This was prepared by adding so long dilute milk of lime to double superphosphate until the acid reaction had just disappeared, 356 T. Katayama. carbonate! was again clearly demonstrated, the effect was very different with a soil containing 119¢ humus, as shown by the foregoing experiments of S. Suzuki. No depression was noticed in that case by moderate liming. 1 This could, in my sand culture experiment, be due only to the neutralization of the acids of the roots. Ueber den Kalkgehalt verschiedener tierischer Organe, IV. VON M. Toyonaga. In meinen friiheren drei Mitteilungen! habe ich nicht nur den Kalk- und Magnesia-Gehalt in verschiedenen Organen nach meinen Unter- suchungen mitgeteilt, sondern auch darauf hingewiesen, das der absolute Kaikgehalt der Driisen bedeutend grosser ist als derjenige der Muskeln und der weissen Hirnsubstanz. Nur bei den Hoden hat sich eine weit geringere Kalkmenge ergeben als bei anderen Driisen. In dieser Mitteilung handelt es sich um den Kalkgehalt der Leber von Pferd, Rind und Schwein, ferner um den der Schilddriise des Pferdes. Fiir die Schilddriise existieren noch keine derartigen Bestimmungen, wihrend fiir die Leber vom Menschen und Hund solche bereits bekannt sind. Fiir die Leber hat Oidtmann in 1000 Teilen frischer Substanz 0,2842 Teile Ca und o0,o125 Teile Mg gefunden, waihrend Aloy? in 1000 Teflen frischer Hundeleber 0,175 bis 0,259 Ca und 0,048 bis 0,066 Teile Mg fand. Meine Bestimmungen wurden fiir die Schilddriise der Pferdes ebenso ausgefiihrt wie in meinen friheren Mitteilungen erwahnt, mit folgendem Resultat : SCHILDDRUSE DES PFERDES. | CaO, MgO. Frische Winker ‘Trocken- eta (in 1000 Teilen (in 1000 Teilen Ca. Substanz. ea 7 23 substanz. eres frischer frischer Mg. Substanz.) Substanz, 67.508 ¢ | 39.0586 g 28.4404 ¢ 0.4818 ¢g 0.3517 % 0.116 1.85 57-858 % 42.142 9% | 7-137 Jn 1 Diese Bulletin, Bd. V u. VI. 2 Jahr. Bericht f. Tierchemie Bd, 32. S. joo. 58 M. Toyonaga. Oo Was die Lebern anbetrifft so wurde hier das Wasser-extrakt separat untersucht, wahrend das unléslich Gebliebene mit Essigsaiure von 1% extra- hiert und hierin auch Kalk und Magnesia bestimmt wurde. Das unlésliche wurde zuletzt mit Alkohol (93%) extrahiert und auch in diesem Auszug Kalk und Magnesia bestimmt, Schlieslich wurde der Rickstand ebenfalls auf Kalk und Magnesia untersucht, Das Resultat ist aus folgender Tabelle zu ersehen: PFERDELEBER. Trocken- é | substanz. peo es ee = ig grams. g =e § 2 § Se Mg. Wasser-Extrakt ......... 17.300 0.925 0.0103 0.0248 0.5 Essigsiure-Extrakt...... 12.800 0.4901 0.0261 0.0264 rz Alkohol-Extrakt ......... 6.700 0.2076 0.0050 spur Riickstand: .2.0.::.0...- "estos 0.1968 spur 0.0028 PVCU TMeimeeele was dectedestene | 52-775 1.8195 0.0414 0.05417 0.9 In 1000 Teilen frischer | Substanz...a-s-.-.ss0s< |. 263:875)% 6.0975,% 0.207 % (0.27085 2% =0.1478%Ca) =0.1681 °4~Mg | RINDSLEBER. cc ne emmmmmmmmmmn tmmmmmanemmmmcemnmseesmeenenens: cuemeseemnn semen eens ‘Troken- : “s subeiaee, Asche. | Cat ), Mee Ca, grams, grams, grams. grams, | Mg. ——____|______|— 3 a r wb Wasser-Extrakt ......... | 21.070 1.0568 0.0202 0.0336 0.7 | | | | Essigsiiure-Extrakt...... 8.200 | 0.7028 | 0.0242 0.0260 | I. Alkohol-Extrakt ......... 5-350 0.1140 0.0048 0.0024 2.4 Riickstand .....ccessaen 12,0056 0.0538 | 0.0045 0.0035 | 1.5 ; MIMS a das ehavaenicu sees 42.6256 1.9274 0.0537 | 0.0655 1.0 In 1000 Teilen frischer | SBSH Sosssasee eck 233-128,%0 9.637,.% | 0.2685,% 0.3275.% =0.1918% Cal =0.1977,.%Mg EEE EEE Ueber den Kalkgehalt verschiedener tierischer Organe, IY. 359 SCIIWEINSLEBER. ken- : Trocke Asche, |. CaO.: | + MgO. Ca. substanz. yrams grams grams ~ Meo. grams. Sy Babi een San Mg. | Wasser-Extrakt ......... 26.560 2.3544 | 0.022 0.0356 07 Essigsaure-Extrakt...... | 12.280 0.6018 0.0209 0.0223 1.1 Alkohol-Extrakt ......... 4.750 0.083 0.0043 spur PGSM oc... ccsse2 9.863 | 0.0446 0.0026 0.0035 0.9 PAULL as ys-ccieaccces cates Bo.643,° || 3.0348 | 0.0498 0.0614 1.0 In rooo Teilen frischer SetaNZ yes cuspe | eZOZ TES 15-441% | 0.249% | 0.307% = 0.1779 4003) =0.1853,%Mg Wahrend im Wasser extrakt die Magnesia tiber den Kalk iberwiegt, sindim Essigsaure extrakt keine wesentlichen Unterschiede zu bemerken Ob die geringe Menge Ca und Mg im Alkohol-extrakt vielleicht auf Spuren von Ca-und Mg-Seifen zuriickzufihren ist, ware wohl einer weiteren Priifung wert. Vergleichen wir den Calciumgehalt dieser drei Lebern mit demjenigen der Hunde- uud Menschenleber, so finden wir eins ziemlich gute Ueberein- stimmung, namlich ftir 1000 Teilen frischer Substarz : | wae Me. FTAs CR Ee, ee dear cin owtakesetieenet | 0.175-0.259 0.048-0.065 IIGHSEHERICDED oc. css. 7 ; at On the Flowering of Bamboo. BY Oscar Loew. . To those agricultural plants, the flowering of which is considered asa very undesirable feature by the farmers, belongs the bamboo, since after flowering and fruiting the bamboo dies off like other Graminee. There may pass 20 to 60 years, however, before the flowering takes place and during these many years the bamboo groves forming so often an essential part of a Japanese farm, has produced annually innumerable shoots from the extensive system of rhizomes, thus increasing considerably the income of the farmers. These shoots are cut when 20-50 cm. high and sold in the marke.t They form a much esteemed, popular, dish and are prepared in various ways. Of the varieties growing in Japan it is especially Phyllostachys mitis that yields the most palatable shoots. David G. Fairchild! writes on the bamboo as follows: ‘The bamboo groves of Japan are not only one of the most striking features of its landscapes but one of its most profitable plant cultures. No other nation has found so many artistic uses for the plant as the Japanese and in no other country, except it be China, is such a variety of form employed by the common people. The plant is a necessity to the Japanese peasant ; it forms one of the favorite themes of the Japanese artist and out of it are manufactured some of the most delicate works of Japanese art. 1 Bul. No. 43 of the Bureau of Plant Industry, Washington. Mr. 3. Zathrep has warmly recom- mended the introduction of the Japanese bamboos into the United States. Very valuable studies on bamboos have been published by Prof. 7. Makino, 366 Oscar Loew. The bamboo is in fact one of the greatest cultivated plants of this plant- loving race.”’ . It can easily be perceived that it is considered as a great calamity when an extended bamboo grove commences after many years of existence, to develop flower.!. Climatical conditions, as very warm and dry summers, also the age of the rhizome and perhaps the gradual exhaustion of the soil when manuring is not properly attended to, may play a role. The simplest way to prevent the evil would seem to cut away all the buds as soon as they appear. But with these tall plants densely crowded in eroves this would involve too many difficulties, which have also been en- countered with the flowering of the sugar cane, also a dreaded phenomenon. The possibility however cannot be denied that under certain condition of nutrition the leaf and shoot formation may be so much favored that all organic nutrients are again consumed in this process, not sufficient being left for development of flowerbuds. It is well known that on different soils and with different manuring the ratio of straw to grains varies between wide limits with the Gramineze. Although some influences acting in this direc- tion are known, a proceeding to prevent the flowering altogether in a well developing plant is not yet known. What may be gathered from the litera- ture in this direction is about the following : Gypsum acts more on leaves and stems than on grains.?. Blomeyer® observes: ‘Gips lisst die Vegetation, die er offenbar sehr begiinstigt, bisweilen nicht zum Abschluss kommen. Die Bohnen héren nicht auf, an der Spitze zu griinen und zu bliihen, sehr zum Schaden der Ansatzes uud Ausbildung der Friichte am unteren Stengelteile.” Also a rich manuring with sodium nitrate seems to act more on leaves than on flowers, as //. Miiller4 observed with potatoes and sugarbeets ; the starch was rapidly transformed into protein, the leaves assumed a deep 1 Of other late flowering plants may here be mentioned Agave America, Larix europea (20-30 years), Pinus and A/nus (12-40 years, according to climate). 2 EE. Wolff, Praktische Diingerlehre, Berlin, 1892. % Die Cultur der landwirtschaftlichen Versuchspflanzen I, p. 330. : 4 Centralbl. f. Agriculturchem, 24, p. 454; also Chem, Centralbl. 95, II, p. 682. On the Flowering of Bamboo. 367 green and contained 2.5 times as much chlorophyll as the check plants, but the formation of flowers was retarded. Schnetdewind' reports that potassium nitrate acts with cereals more on the development of leaves than on that of the grains, while it is the opposite with magnesium nitrate. Magaoka? however did not observe such an effect of magnesium nitrate on rice. Wilfarth® found that manuring with little potassa and much nitrogen induced the sugarbeet to develop a rich leafy vegetation, the roots however remained poor in sugar, while under the opposite condition the roots ac- cumulate much sugar. When the amounts of phosphoric acid and nitrogen are reduced at the same time the plants will remain very small. It has further been repeatedly observed that rich phosphatic manure leads to an early, Chilisalpeter to a late ripening. Also different potassium salts seem to have an influence on the rapidity of flowering. Several reasons led the writer to suppose that an excess of lime and nitrogen over the other mineral nutrients should favor the growth of leaves most, to the detriment of flower formation. An experiment made in this direction led however not to a quicker leaf-formation but to a very marked increase of the size of the leaves. A barley plant which had been grown in the following culture solution : (2 UE SES oa, Meremestan SUlphate 7.0..2........0..00..05.. GENS; Monopotassium phosphate .................. E> 1g Reeeesoieiy Mithaue 0.) ei. ee OZ 5; Lis dolsisliecjs 0 - > <0. i, QT ,; and had developed three large and two small stalks, was at a height of 29 c.m. transferred into the following solution (Febr. 15) Jog Cath: ae | RE 0.5 % oe eu GR) UNIT) fo 9 A 0.01 ,, Prmmonium SUlpMate wn, 0.k ec cee cana ces 0.10 ,, 1 Journ, f. Landw. 1808. 2 These Bulletins VI, No, 3. 3 Zeitschr. f. Zuckerindustrie, 51, p. 323 [1901]. 368 Oscar Loew. Monopotassium phosphate .................. 0.10% Potassiuin nitrate 2... 2.42222 eee O20% Ferric phesphates. ..ctie4iis Ao OPE aes The amount of magnesium sulphate was here exceedingly small in order to make the effect of the excess of lime more clearly appear. Since some more magnesia had very probably been absorbed from the first culture solution, than absolutely needed, further growth to a certain stage could be expected ; indeed within the next five weeks the height increased to 48 cm. However, the root and the lower leaves had stopped growth, it were only the youngest leaves that had grown and their growth in length and width continued three weeks longer until they had reached April 15, a quite unusual size as will be seen from the photograph, reproduced on Plate XXVII. The measurements of the two largest leaves were as follows : Length 30 and 31 cm.-—Checkplant............... 20 and 22 cm. Width 3.6 and 3.6 cm. TT reer 1.5 and 2:1 (Gar Soon afterwards a gradual yellowing set in and the examination of the roots showed that they had partly died off, leading to an early death of the entire plant. The cause was very probably the prevention of assimilation of phosphoric acid and the precipitation of the soluble phosphates of the protoplasm as calcium phosphate. Manuring experiments with bamboo with the intention to check the formation of flowerbuds would naturally require a long series of years. Ob- servations, however, made on annual grasses may justify a heavy application of gypsum and chilesalpeter in conjunction with irrigation when groves are suspected to soon develop flowerbuds, which is to be feared especially in very warm and dry summers. The question of retarding flowering and fruiting naturally suggests the further question whether the life of the leaves of the flowering Graminee might not be prolonged beyond the usual duration. The ripening of the grains depends here upon the death of the leaves which gradually turn yellow from the lowest to finally the uppermost. Since the seeds require relatively much dipotassium phosphate it is probably this salt, so necessary for the protoplasm of every cell, which is drawn chiefly from the leaves to a re, On the Flowering of Bamboo. 369 the forming seeds thus causing the death of the leaves.1. Heinrich observed that even the root becomes finally exhausted. This process has its analogy in the migration of a certain amount of potassa, magnesia, phosphoric acid and nitrogen from the deciduous leaves in autumn into the bark of the branches and trunk for later use again. Hence the idea suggest- ed itself whether a dose of dipotassium phosphate applied as top dressing just after the flowering period would act beneficially in prolonging the life of the leaves which thus could produce more starch. The result would be larger and heavier seeds. I have made in conjunction with Prof. K. Aso several experiments in this direction but the results were not decisive, although we have not only applied single salts (6-10 g. per pot of 8 Kilo soil) but also complete nourishing solutions as top dressing after the flower- ing period. In some cases the leaves did not die off as rapidly as in others and also the weight of 100 grains was a little larger than with the check- plants, but these differences were too insignificant to bear any weight.? 1 According to Wolffand to Ritthausen it is one of the offices of SiO, to hasten here this dying process in favor of the seed. 2 Deficiency of nitrogen enhanced the yellowing process, a certain excess of lime or of dipotassium phosphate retarded it. With KCl the green was a little longer preserved than with K,SO, as a potassa manure, Also a certain excess of water retards the yellowing. eS SS eee BULE. COLL, AGR., VOL, VI PLATE XXVII Growth of Leaf caused by an Excess of Lime. To) Bamboo, eee Ya | ie lee + / vs Further Observations on Oxidases. kK. Aso: In order to furnish further proofs that the substance contained in plant- juices which produces the guaiac reaction is not identical with a substance contained in some plantjuices that liberates iodine from potassium iodid, I have made some further experiments to show that the latter is merely a nitrite. In the first place, the following observation was made in order to test the assertion of Bach and Chodai,! that the guaiac reaction upon peroxids is more sensitive than the liberation of iodine by peroxids. The common paraldehyd of commerce has generally an acid reaction and yields with potassium iodid-starch very soon an intense blue reaction due to the liberation of iodine. I entertained the supposition that this reac- tion is not caused by the pure paraldehyd, but by an admixture of an organic peroxid, very probably by acetylhydroperoxid.? Similar peroxids have been observed as a result of autoxidation of other aldehydes and also the common ether forms after long contact with the air, an organic peroxid, which sometimes even causes explosions in distilling such an old ether to the last drop. ‘I shook therefore about 20 c¢.c. of commercial paraldehyd with an equal volume of 10% sodium carbonate solution and after washing until the alkaline reaction disappeared, a portion of that paraldehyd was distilled off. There was now observed that the iodine reaction above-men- tioned did not take place neither at once nor within fifteen minutes, but only 1 Berichte der D. Chem. Ges. 1904, XNXVIT. Heft 1. 2 R. H. Page. (Amer. Pat.) mentioned in the Chemiker Zeitg, Benzaldehyde (commercial) pro- duced also the iodine reaction in traces. 372 K. Aso. an exceedingly weak reaction slowly appeared later on, which however was not intensified by the addition of some acetic acid. The original paraldehyd, however, gave an intense reaction within a few minutes. This result was sufficient to prove that it is not the paraldehyd itself, which causes the iodine reaction, but some impurity, which can only have been the peroxid above-mentioned, to judge from analogy. Now it was interesting to observe that the original paraldehyd which produced such an intense iodine reaction had no reaction whatever on tincture of guatacum, not even on addition of some hydrogen peroxid. These mixtures were still colorless even after half an hour. Therefore I can not agree with Bach and Chodat when they believe ,, dass die Guajakreaction auf Peroxyde bei weitem empfindlicher ist als die Jodkalium-Stirke-Reaktion.” Also in regard to nitrites, both reagents were compared with the result that the guaiac reaction is less delicate than the potassium-iodid-starch reaction.!. Most of plant juices produce very strong guaiac reaction, but no potassium iodid reaction. Hence the substance which produces the guaiac reaction must be quite different from that which produces the potassium iodid starch reaction, that is, the former is caused by oxidase very frequently in plant juices, and the latter by nitrite which is present in certain plant juices, as I had positively proved in one case. But if the iodine liberation by certain plant juices would be due always to traces of nitrite and not to enzyms, how is the fact to be explained that this property is lost in most cases on heating ? The probable answer is here that plant juices are often slightly acid and contain at the same time small quantities of amido-compounds. Under this condition traces of nitrites must disappear on warming, while after addition of some alkali, the reaction will probably be maintained after boiling. 30 c.c. of 0.001% potassium nitrite solution were mixed with 30 c.c. of 19% asparagine solution and divided into three parts. To one, was added a drop of dilute acetic acid, to the other a drop of dilute caustic potash solution, while the third served as control. 1 For instance, with the solution of 0,0005%% potassium nitrite, a distinct iodine reaction appeared ’ immediately, but no guaiac reaction at once and only a trace after half an hour, Further Observatious on Oxidases. 373 These solutions were kept boiling for five minutes and tested with potassium iodid starch with the following result : Control. Alkaline solution. Acid solution, The reaction appeared, but Distinctly : ean ; 1 ; : 7 No reaction at all slower and weaker than in and rar eer : : after several hours. the alkaline liquid. immedialy. iret This experiment was repeated several times with the same result. Hence I became convinced that amido-compounds decompose nitrite in a very faint acid solution and it is necessary to make the solution alkaline to preserve the nitrite. Thus I made analogous experiments with plant juices. 18 grams of the buds of Sagittaria were crushed, extracted with 100 c.c. water and divided into three equal parts. Toone, a few drops of acetic acid, to the other a few drops of caustic potash were added while the third served as control. Each solution was heated to 95° C. for 10 minutes and filtered after acidification with acetic acid, which had produced some pre- cipitate, and tested : | Control. Alkaline solution. Acid solution, | —_— —__ ——_ No reaction at first, but . : dns No reaction at first, but after 10 min., a reaction Potassium-iodid ei Aare ; : 7 aie ° after 10 min., it appear- Distinctly at once. appeared although weak- starch reaction. : oo Ra ae ite Ree ed gradually. er than in the control case, Griess reaction. Distinctly. Distinctly. Distinctly. Guaiac reaction. No reaction at all. Ne reaction at all. No reaction at all. In order to separate the substance which produces the guaiac reaction from that which yields the reaction of Griess, the following experiments were made: 35 buds of Sagittaria (about 33 grams) were crushed with 55 c.c. water. To 60 c.c. of the pressed juice which yielded a very strong reaction with potassium-iodid-starch, 200 c.c. of strong alcohol (90%) were added. The mixture was left for twenty four hours and filtered. The filtrate was evaporated on a waterbath and the residue was dissolved in 374 K. As). 20 c.c. water and filtered. The filtered liquid gave a strong Griess reaction as well as the iodine reaction very decidedly, but not the guaiac reaction while the aqueous solution of the well-washed precipitate gave, in the con- trary, not the Griess reaction nor the iodine reaction, but a strong guaiac reaction. This result proved positively that the substauce which gives the guaiac reaction ts not the same that liberates todine from potassium todid. Similar experiments were repeated with the bud and .the skin of the bulb of Sagittaria and the same results were obtained in each case. Ex- periments with buds of potato were also made with similar results. ! CONCLUSION. 1. The guaiac reaction for peroxids is not so sensitive as the potassium- iodid-starch reaction, and the guaiac reaction for nitrites is much weaker than the iodine reaction for nitrites. 2. The reason why certain plant juices which can liberate iodine, lose that property on heating is very probably due to the acidity of the juice and the presence of traces of amido-compounds, which are very favorable conditions for the decomposition of nitrites. 3. It was positively shown against the assumption of Bach and Chodat, that the substance which gives the guaiac reaction is not the same as that which liberates iodine. 1 A more detailed report on this subject will appear in Beihefte Bot. Centr.-Bl. a On the Large Bacillus observed in Flacherie. BY S. Sawamura. _ ‘Since Pasteur had demonstrated the presence of a large bacillus in the intestinal canal of the silk-worm suffering from flacherie, it has been the subject of much discussion. Macchiati! gave the bacillus the name of Bacillus bombycis, while the writer? and Lo Monaco? regarded it as Bacillus megaterium De Bary. It seems to be also the same as the bacillus, whose pathogenic activity was first opserved by Jshiwata, and is called sometimes by the name of “‘ Sudden Death Bacillus.” By further investigations on this subject the writer found that the large bacillus usually seen in a large number in flacherie has slightly different culture-characters from Bac. megaterium De Bary ; that is :—the bacillus in question produces the indol reaction which is usually wanting in culture of Bac. megaterium ; and the colony on agar seems to grow larger than the latter. The question, whether this bacillus may be regarded as a distinct species, or a variety of Bac. megaterium, must be decided by further inves- tigations. But for the sake of convenience we shall call this bacillus, for the present, by the name of Bacillus megaterium bombycis. Experiment J. This cxperiment was performed to observe the pathogenic action of Bac. megat. bombycis. 1904, May 10. The following materials were fed 1 Le Stazioni sperimentali Agarari Italiane Vol, NX, Part II. we Bulletin of Agricultural College, Tokyo, V. No. 4. i] Dall’ Archivio di Farmacologia e Scienze affini. Year IL, Vol. Il, Part VI-VII. 376 S. Sawamura. together with mulberry-leaves respectively to 20 silk-worms on the forth day of the second stage. 1. Six days old agar-culture of Bac. megat. bombycis suspended in water. 2. The fluid I. to which a few drops of dilute acetic acid were added. 3. The fluid I. diluted with 10 times its volume of water (average 5 bacilli in an eye-field of 800 fold magnification). 4. Pepton-glucose culture of Bac. megat. bombycis (10-15 bacilli in an eye-field of 800 fold magnification). The larvae fed with an agar culture of Bac. megat. bombycis lost appetite, vomited a yellow fluid and died. Vhe dead body shrunk,! and many of the large bacilli multiplied in the digestive canal, but the appearance of the excreta was normal. The number of the dead was as follows :—— Date of observation. Control. le | II. Tie PWe May 11, 3 p.m. fy AGES? 5 ” 14, 9 ” Total. FIZ Od. I‘rom these figures it will be seen: 1. That itis certain that Bac. megat. bombycis exerts a pathogenic action on silk-worm. bo That when Bac. megat. bombycis had served in a small number, however, the pathogenic effect was greatly decreased. 3. That Bac. megat. bombycis cultured in fluid media showed no pathogenic action. 4 With o!'d larvae the dead bodies stretched. On the Large Bacillus observed in Flacherie. Experiment 11, This experiment was performed to observe a second time whether Bac. megat. bombycis loses its pathogenity by being cultured in fluid media. May 14. The following materials were fed respectively to 10 larvac in the second stage. 1. Agar-culture of Bac. megat. bombycis suspended in water. 2. Five days old bouillon-culture of Bac. megat. bombycis (5 bacilli in an eye-field of 800 fold magnification). The number of the dead within 5 days was as follows : — Control oO li 10 Jue fe) The results of this experimeut shows again that Bac. megat. bombycis loses nearly all its pathogenity by being cultured in fluid media. Why the bacillus cultured in fluid media is less pathogenic, only further investiga- tions can decide. Experiment 11. This experiment was performed to confirm again that Bac. megat. bombycis does not injure silk-worms when inoculated in a small number. May 18. The following materials were fed respectively to ro larvae on the third day of the third stage. 1. Agar-culture of Bac. megat. bombycis suspended in water. 2. The above fluid diluted with 10 times its volume of water (average 5 bacilli in an eye-field of 800 fold magnification). 3. The fluid I. diluted with 50 times its volume of water. 4. The fluid I. diluted wilh roo times its volume of water. Two series of the experiments were performed, and one series was kept at 25°C. and the other at 18.99 C. The number of the dead was follows :— as 378 S. Sawamura. mT | Date of |Temperature.| Control. | L IL. TI. IV. observation. | May 19, 9 a.m. a5e'C. : fe) 71 Z fe) fe) 39 20, 7 7? 2 te) 3 7 5 Le) » 21,5. » ” 12] =; @) 2 I Total. fo) 10 | 9 i I May 19,9 a.m. 18.9° C. ° 4 I | o oO 93 20,5) 159 | ” 18) 6 6 Oo oO 9» 21,55 95 PY | 12) == Z 2 oO Total. fe) 10 | 9 2 ° We may conclude from these results. 1. That when Bac. megat. bombycis is inoculated in a small number the pathogenic effect is greatly decreased. That higher temperature increases the pathogenity of Bac. megat. 1S) bombycis. These facts may explain why flacherie was regarded as terribly infec- tious by Lo Monaco,! while as quite harmless by Bolle,? the cause of the difference probably being the quantity of the bacillus fed and the tempera- ture in which the trial larvz were kept. Experiment IV. This experiment was performed in order to observe the action of Bac. megat. bombycis on animals other than silk-worm. May 12. Dasychira lumulata Butl. was fed with a large quantity of agar-culture of Bac. megat. bombycis. On the next day one of them was killed and the multiplication of the large bacilli was observed in the intestinal canal. The remainder were kept for 5 days without observing any symptoms of the disease. * Dall’ Archivio di Farmacologia Sperimentale e Scienze affini, Year I, Vol, II. Part VI-VII. * Bericht tiber die Titigkeit der K, K. Landw. chemischen Versuchsstation in Gérz, 1902. On the Large Bacillus observed in Flacherie. 79 o>) In another experiment 0.1 c.c. of a water suspension of Bac. megat. bombycis was injected into the intestines of Dasychira lumulata through the anus by a specially constructed syringe. They all died on the next day. The leave-fragments in their intestinal canals were brown and the alkality of the intestinal fluid was weaker, and many large bacilli were detected therein. June 23. Ten larvae of Gastropocha pini L. were fed with an agar- culture of Bac. megat. bombycis from which two died; but the cause of death seemed to be another than the infection with the bacillus, because it was not detected in the intestinal canal of the diceased. The remainder were quite healthy. Five of the other larvae were inoculated subcutane- ously by stabbing softly with a sharp needle holding Bac. megat. bombycis onthe point. On the next day 3 of them died, in whose blood the large bacilli had multiplied. On the following day one more died, but the last formed a cocoon. Two white mice were fed with a large quantity of agar-culture of Bac. megat. bombycis, and one was subcutaneously inoculated, both showing no symptom of disease. From these facts it may be seen: 1. That Bac. megat. bombycis is pathogenic also for other insects, although they have stronger resistance-power than the silk-worm. 2. That Bac. megat. bombycis is not pathogenic for mammalia. Experiment V. This experiment was made to decide whether Bac. megat. bombycis produces any poisonous substance. The difficulty in this experiment lies on the fact, that as Bac. megat. bombycis adheres always to mulberry leaves, it multiplies very soon in the intestinal canal of the silk-worms, when their health is injured in any way, such as feeding a sterilized bacterial culture. To obviate, therefore, any errors in this direction mulberry-leaves sterilized with formalin served. The feeding of the filtrate of the bouillon culture passed through Chamberland’s filter was without effect on silk-worms. 380 S. Sawamura. May 30. Twenty silk-worms on the first day of the fifth stage were fed with agar-culture of Bac. megat. bombycis sterilized with a few drops of formalin, and afterwards reared with sterilized leaves. Other 20 larvae were also fed with sterilized leaves as a control. On May 31 two and on June 1, one of the experimental larvae died, in whose intestinal canal no bacillus was detected, the leave-fragments being green, whilst no death took place in the control larvae. The average live weight of the larvae on June 1 was as follows :— Experimental larvae 0.790 grams. Control fe L.240-. 3) Difference A G:550 < 5 After June 1 natural leaves served from which 4 of the experimental larvae died, in whose intestinal juice the large bacilli propagated. From these results it is very probable that Bac. megat. bombycis produces some substance strongly poisonous for silk-worms. Experiment IV. This experiment was performed to ascertain the distribution of Bac. megat, bombycis. 1. May 2. Silk-worms, which had just been hatched from the eggs and were not yet fed, were washed with strong alcohol, and the alcohol adhering was burnt in order to sterilize the larvae, They were then cut with sterilized scissors and put into bouillon. Among 3 tubes of bouillon 2 remained clear, but one became turbid, whence plate-cultures were prepared. From these plates a large bacillus was isolated, which was proved to be Bac. megat. bombycis by killing 5 out of 10 larvae when the bacillus was fed to them. to Bac. megat. bombycis was also isolated from the excreta of a young larva fed 16 times with mulberry-leaves, and then it was fed to 10 larvae, whieh died from this within five days. — On the Large Bacillus observed in Flacherie. 381 U2 This experiment was repeated with the same result when this bacillus from the fresh excreta of an older larva in the third stage was used. 4. Bac. megat. bombycis cultured on agar and derived from mulberry leaves directly, killed 8 out of 25 larvae within 4 days by feeding this culture. 5. Dasychira lumulata died within a few days when it was injected with pure water through the anus. In the intestinal canal large bacilli always were observed which were proved to be Bac. megat. bombycis by its pathogenity to silk-worm. From these facts it may be inferred that Bac. megat. bombycis is one of very widely distributed bacteria. It exists always in the alimentary canal of healthy insects without causing any injury to them. But when it is cultured on solid media and fed to silk- worms, death soon follows the operation. The peculiar fact can not at present be explained. Further investigations on this point are necessary. Experiment VII. Various other bacteria were fed to silk-worms for comparison with Bac. megat. bombycis. The results were as follows :— 382 S. Sawamura. Number Name of Bacteria. Date. eee of larvae Dead. : ; used. 3 eS j First day of 1. Bacillus isolated from May 10-14. | the second 10 fo) cherry leaves, stage. 2. Bacillus prodigiosus. og or Ife) fo) 3. Bacillus coli from Nukamiso. a aa i c Bacillus coli from dl B a 5 silk-worm, 5. Bacillus II. from mulberry. = ss ie) fo) 6. Bacillus IX. a : 2 yehe ae ° 7. Micrococcus I. B. . ss 5 ” fe) 1°) 8) Micrococcus 1iB:) 5 ay ms op 10 fo} 9. Micrococcus HI. B. ,, - a = 10 fo) 10. Micrococcus VII. B. ,, ne “a a 10 fo) 11. Micrococcus XI. B. ,, = 5 _ 10 fe) Third day of 12. Micrococcus VUI. B.,, " May 18-21. the third 10 fo) stage. Fifth day of 13. Bacillus megaterium,+ June 4-6. the fifth 5 fo) stage. 14. Bacillus coli from - silk-worm, 22 dd 5 Except Bac. coli there were no bacteria which caused the disease of silk-worm by feeding, as Bac. megat. bombycis did. 1 Obtained from Germany, On the Large Bacillus observed in Flacherie. 383 Experiment VIII. The following bacteria were used to observe their pathogenity for sillk- worms by subcutaneous infection. j a | } Number of Name of bacteria. Date. Age. larvae used Dead. | Remarks, ; First day \Numerous large bacilli ly ar oem ie of the 6 6 | in the blood; id acillus. 3- fifth stage. bodies soft and black, 2. Micrococcus ee: +3 » 5 o from mulberry. 3. Micrococcus WAIE TB: ee i = o from mulberry, | 4. Micrococcus WWIII eye a fi is o from mulberry. 5. Micrococcus from healthy ¥ = 5 fo) | silk-worm., 6. Bacillus typhi murium. a 22 5 2 A 7. Bacillus coli from Second day Short bacilli in the pee a Te June 1-4. of the 5 5 blood; dead bodies 5 fifth stage. shrunk, : | Many bacilli i 2 8, Bacillus g ot Goce »yocyaneus, 2% 3 mine DENTS tase pyocyaneus with green tint, a Large bacilli in the : blood ; dead bodies ! : ~ 9. Bacillus S és ee. te 1 black jus i be 5 « + 5 2 soit anc 5b ack just as megaterium. those killed by Bac. | megat, bombycis. 10. Micrococcus » » 5 from mulberry. oO 1 Provided by Mr. Hayashi. Nomura believes this identic with B. alvei. 2 Obtained from Germany. 384 S. Sawamura. Number of Bred: Name of bacteria. Date, Age. laroaeaiseds - Remarks, : Fifth day of 11. Bacillus 1 | June 4-6. the fifth 5 5 Same as 9. megaterlum. Saas 12. Bacillus coli from silk-worm, » ” 5 47 Same as 7. 13. Bacillus coli from >* “"Nukamiso. ” ” 5 4 Same as 7. 14. Bacillus pyocyaneus, 2 a 5 5 eo ot 15. Bacillus acidi lac- tici Hueppe. 2 ad 5 2 16. Bacillus subtilis, f Pe 5 fo) 17. Micrococcus Large bacilli in De e 5 5 I the blood; dead from mulberry. body black, 18. Micrococcus I. from silk worm Seventh day with black spots; Tune 6-8. | of the fifth 3 oO on the fore-part stage, of the body. 19. Micrococcus II. from the same 4 BS 3 re) worm, 20. Short bacillus from the same P - 3 fo) worm, 21. Micrococcus I, Fifth day of from healthy June 7-10, the fifth 5 oO silk-worm, stage. 1 Obtained from Germany. On the Large Bacillus observed in Flacherie. Name of bacteria. iP) ios) 24. 25 . 26. 27. 28. 29. 30. Sr. 32: 33- . Micrococcus V, from healthy silk-worm, . Micrococcus VI, from healthy silk-worm, Micrococcus IT. from sick worm with black spots. Micrococcus IY. from the same worm, Date. June 7-10. 39 Short bacillus from the same worm, Bacillus megaterium bombycis. Bacillus from healthy silk- worm, Micrococcus pyogenes aureus. Bacillus prodigiosus, Sarcina lutea, Bacillus megaterium bombycis from mulberry, Bacillus from sick worm, June 9-13. June 15-17. Number of | Age. evaptaead. | Dead. Remarks, Fifth day of the fifth 3 fo) stage. » 3 o ” 5 oO 7 5 Oo ” 3 oO - 3 3 Same as I, First day of the fifth 5 fo) stage. Many n.icrococci Sixth day of in the blood ; dead the fifth 5 3 body of one hard but stage. two soft; not blackened. ” 3 S ” 5 ca) First day of the fifth 5 5 Same as 1. stage, ” 5 2] 386 S. Sawamura. From these results it follows that many bacteria, such as Bac. megat., coli, pyocyaneus, Micrococcus pyogenes aureus, &c., can multiply in the blood of silk-worms and kill them as Bac. megat. bombycis. As the action of Bac. megat. on silk-worm is the same as of Bac. megat. bombycis, it is very probable that the latter is a variety of the former. GENERAL CONCLUSION. In the former investigations the writer assumed that flacherie is caused not by any special bacterium but by several, which occur commonly on mulberry-leaves. This assumption is also confirmed by the results of these ex periments. The writer expresses his sincere thanks to Mr. Hayashi who provided BI him with a culture of “Sudden Death Bacillus” and to Mr. Yamasaki, Assistant of the College. TEMPERATURE DURING THE EXPERIMENTS. Date. |Temperature.| Date. |Temperature.| Date. /Temperature.| Date. (Temperature. May 1 15.0°C. | May 13 201071 €F |) May 25 12:82 €; | june 6 cabal Cs 2 1222, Pgh Gi II. Suez 15.0 eee? 22.2 a _ Some New Varieties of Mycoderma Yeast. BY T. Takahashi. The “ Kahmhefe” well known by the rapid formation of a covering film on the surface of nutrient liquids occurs very abundantly in Sake, Sake- mash and Koji i.e. boiled rice covered with a vegetation of Aspergillus Oryzz. But the researches hitherto made on this subject are very scanty. Kloécker and Schioning! mentioned the presence of a variety of S. Anomalus in sake-koji, Yabe? observed the presence of a mycoderma yeast on the rice straw used in the sake factories. Kozai cuitivated from koji two kinds of “kahmhefe” viz. a variety of Saccharomyces anomalus with hat shaped spores and an asparogenous yeast, both causing a feeble alcoholic fermenta- tion and producing acetic ether in beer wort. Quite recently A. Sazté isolated from sake a kind of S. Axomalus, which produced acetic and butyric ethers in beer wort or in koji extract and was capable of fermenting dextrose, levulose, saccharose, galactose and sparingly also maltose. As this kind of yeast causes various changes in the constituents of the substrata, in which it grows, it is very important to investigate its character and also the changes which it produces in nutritive liquids. From this point of view, I have isolated several varieties of Mycoderma _ yeast? from sake, koji and sake-mash and studied their morphological as well as physiological properties. They are briefly described in the follow- ing table. 1 Centralblatt f. Bakt. Parasitenkunde. Alth II. Bd. I 1895, p. 777. 2 Bulletin of Imp. Univ, College of Agr. Vol, III, No. 3, P. 223. 3 By mycoderma yeast I mean the kind of Aahmhefe, which is in capable of forming spores. 388 T. Takahashi. MYCODERMA A, FOUND IN KOJI. (Plate XXVIII A). GENERAL CHARACTERISTICS. Behavior Bormiand usual Growth. to ee sugars, Elliptic and sau- On the wort (kept at 22° C.), a thin white mealy Ferments sage shaped, 1°5- | film is formed after 22 hours. The film changes to | glucose,sac- ' 2p. in b. and 12- 15 fie es Rich an vacuoles (4-5), which contain re- fractive and re- volving bodies or granules, greyish brown and assumes a mesenteric structure. The colonies on the wort-gelatine are round chalky white with mesenteric surface. Streak culture on wort or kdji-extract gelatine have a coarse mesenteric appearance; the gelatine is very slowly liquefied. Giant colonies on wort-gelatine are greyish in color and folded especially on the margin; on sugar- bouillon-agar, it gives white mesenteric coatings, charose, but not maltose. Assimila- tes glucose saccharose, maltose. Optimum and killing temperature. Optimum temperature; 1g-—25° C. Killed at a temperature of sors Gaim Io minutes. CHEMICAL BEHAVIOR. Behavior to alcohol. Formation of alcohol and acids, Alcohol (07-1 %) contained in Mayer's and Nigeli’s solution de- void of sugar, is changed into acetic acid, which is gradually further oxidized into CO, and H,0. This phenomenon is also observed in the kGji-extract culture, In sake con- taining 13:29 (16°33 vol. %) of alcohol, no growth takes place. It formed 2°2% of alcohol after standing for 10 days in wort at 21- 26°C, 3°35% alcohol, O:199% of fixed! and o*201% of volatile acid we reformed in kdji’s-extract after 1o days culture at 19-26° C., while after 15 days under the same condi- tion were found only 0'024.% fixed and 0'057 % of volatile acid. Formation of CHO. CH. OF was found in kOji- extract culture. Assimilation of nitrite, glycerin and alcohol. Assimilate glycerin and alcohel. 1 The fixed acid was calculated as succinic acid, the volatile acid as acetic acid. 2 As inferred from the reaction of Cazeweuve and Cot/on and by the formation of anilin-vioilet. This small amount of methyl alcohol is probably an oxidation product. & Nitrite was given in nutritive solution containing glycerin instead of sugar, not take place when the solution is made distinctly acid by acetic acid (1 per mile). poisonous action took place, Assimilation can Here evidently a Some New Varieties of Mycoderma Yeast. 389 v MYCODERMA B (Plate XXVIII B) WAS FOUND IN MOTO-MASH. GENERAL CHARACTERISTICS, Form and usual size. Growth. Behavior to Sugar, Optimum and killing temperature, Oval or elongated but not filamental as in mycoderma A. 3-5 pm. long, 2-3. broad. The revolving granules in vacuole are rare. colonies waxy and finely striated. On wort or kdji-extract it forms a yellowish brown corrugated film often with a reddish tinge. colonies on the wort-gelatine are rounded, chalky or a loose mass, Streak cultures on wort-gelatine are mealy but Ferments glucose but granular somewhat concave; on k6ji-extract gelatine tine very slowly on the margin, Giant colonies on wort-gelatine are elevated toward the centre and decorated with concentric rings, and radiating lines. The margin is almost smooth, On sugar-bouillon agar it forms chalky white mesenteric giant colonies. Liquefies gela- | | not saccha- rose or mal- similates these three sugars. Optimum - temp. 26° C. 56> for 10 m, is not sufficient to destroy the | vitality; it is killed at 60° C, CHEMICAL BEHAVIOR. Behavior to alcohol. Formation Alcohol (c"7-1 9%) contained in Mayer’s and Nageli’s solution, de- void of sugar, was assimilated but not converted into volatile acid. In sake with moderate contents of alcohol growth was observed, When the amount of alcohol reaches, how- ever, 10°7—13°23%, no development takes place. Oxidation of ,alcohol took place in k6ji-extract culture. Formation of alcohol and acids. of CH,. OH In wort or kOji-extract after 10 CH,.OH days 2°2% of alcohol, and in the latter media 0°028% of fixed and 0:0095% of volatile acid, while in 15 days 1°8% of alcohol, o°129% of fixed and 070185 % of volatile acids. After 15 days was found 1°8% of alcohol in the wort culture. was found in k6ji-ex- tract cul- ture. | Assimilation of nitrite, | glycerine and | alcohol. . | Assimilates | nitrite glyce- rine and al- cohol. This mycoderma differs from mycoderma A, by having no filamental y y s form, not fermenting saccharose, developing on sake and assimilating nitrite: 392 VT. Takahashi. MYCODERMA C (Plate XXVIII C), ISOLATED FROM KOJI. GENERAL CHARACTERISTICS. Form and usual size. Growth. Behavior to sugars, Oval, elongated saussage or clup- shaped and rich in vacuoles after con- taining revolving refractive granules. Oval 25x35 [., elongated 1-449 f., Saussage and clup-shape 1°4x 12 L. On wort or kdji-extract kept at 22 or 26°5° C. no film was formed but on the latter kept at 28-30° C. there was soon a formation of a film, Therefore this mycoderma differs from two preceding species being only capable of forming a film at higher temperature on the above media. On Heyaduckh’s solution its forms a film also at lower temperature (15-22° C.). On the wort-gelatine it forms mesen- teric waxy colonies. Streak cultures on the wort- gelatine give coarse pasty colonies; on kdji-extract gelatine greyish, firmly corrugated colonies. The gelatine is very slowly liquefied. Giant colonies on wort-gelatine are white mesenteric, with irregular margin; on sugar-bouillon-agar white radiated and folded colonies, Ferments only glu- cose. Assi- milates glu- cose, sac- charose but not mal- tose. Optimum and killing temperature, Optimum temperature 28—30 2 Ge The cells are killed at 544° GC. in 10 Minutes. CHEMICAL BEHAVIOR. Behavior to alcohol. Formation of alcohol and acids. Alcohol (1 9%) contained in In koji-extract 2°78 of alcohol Mayer’s solution, devoid of sugar, | and 0'155% fixed, ‘00095 % volatile was converted into acetic acid. In | acid were formed at 19-26° C. after sake containing 13-39% of alcohol no | 10 days; while after 15 days there growth takes place. were found 2°6770% alcohol, 0°034.9% fixed and 0'032% of volatile acids, chiefly formic and butyric acids, In wort at 21-25° C, after 10 days, was found 275% of alcohol. Also the formation of acelic acid from glycerin, which is contained in Hayduck’s solution, devoid of sugar, Formation GH Ob was found in the cul- ture of kOji- extract- Assimilation of nitrite, glycerin and alcohol, Assimilates alcohol, gly- cerine but not nitrite. a This yeast differs from the preceding species by the higher optimum tem- perature and the formation of acetic acid from glycerin and by varying shapes. Some New Varieties of Mycoderma Yeast. MYCODERMA YEAST D (Plate XXVIII D), ISOLATED FROM KOJI. GENERAL CHARACTERISTICS. 391 Form and usual size. Behavior Optimum and Oval, elliptic, sometimes wedy'e- shaped, ich in vocuoles containing refractive granules of fatty nature, 2-315, > b.,) 3-7 Be We k6ji-extract gelatine have a mealy appearance. The | reddish tinge. cells. The color of the film is in the beginning chalky white (on k6ji-extract) or white (on wort), but after- | wards changed into greyish yellow, often with a On wort-gelatine it forms a round, chalky white colony. Streak cultures on wort or on gelatine is very slowly liquefied. Giant colonies on wort-gelatine are browish yellow, remarkably elevat- ed toward the centre and highly corrugated, on sugar-bouillon agar have a feather like appearance. | cose. Assi- . milates glu- cose, charose, maltose. sac- Growth. to killing sugars. |temperature. ss | —|\— On wort or kdji-extract forms a strongly develop- Ferments | Optimum ed, highly corrugated film, consisting chiefly of ova/ | only glu- | temperature 25-30° C. ~~ = .) 2) ™~ ~ CHEMICAL BEHAVIOR. Behavior to alcohol. Alcohol(1%) contained in Mayer’s solution, devoid of sugar, is changed into acetic acid. ‘The same pheno- menon was found in kd6ji-extract culture. In sake containing 7 3° 32% of alcohol kept at 25° C. 7m _forma- tion took place. After 13 days there was found only 2°28 of alcohol. Formation of alcoholjand acids. Formation of Cr. Ob. Assimilation of nitrite, glycerine and alcohol. In kdji-extract kept at 19-26° C. was found after 10 days 2°39% and after 15 days 5°44% of alcohol and in the latter case o70911% of fixed, and 0°01422% of volatile acids were found. Among volatile acids, acetic and butyric acids were present. In wort kept at 21-26° C, after ro'days was_found'3°29 9% of alcohol. CH,.OH. | was formed | tract cul- } ture, | in kOji-ex- | | Assimilates ' | all three compounds. Thus this yeast is distinguishable from the wedge-shaped cells, by the production of a higher percentage of alcohol (5°44%%) and the growth on sake containing a higher percentage of alcohol (13°39) 392 T. Takahashi. MYCODERMA YEAST E (Plate XXVIII E), FOUND IN KOJl. GENERAL CHARACTERISTICS. 1 | Behavior |Optimum and Form and el Growth. to killing size, | sugar. temperature. Round, oval shap- On kdji-extract forms slowly a thin film, On wort Ferments Optimum ed or elongated, | at first white and thickly foulded, altering in color to glucose, sac- | temperature sausage, often | gold yellow afterwards. Colonies on the wort-gelatine charose but | 22—24° C. wedge-shaped, The | or kdji-extract-gelatine it forms chalky white mesen- | not maltose. | On heating elongated cells are especially rich in vocuoles containing revolving refractive granules. 1°2-1°5 p. b. 1°4-9°4 pw. 1. teric colonies, of which the color changes gradually to greyish-yellow. Giant colonies on wort-gelatine are white with a yellow tinge and highly corrugated. Those on sugar- bouillon-agar are slightly brown and more or less granular in the centre, while toward the periphery radiating lines;run regularly. In addition concentric rings occur on the surface. Assimilates the former two, but not maltose. 55°4 C. for TO” me as sufficient to kill the cells. jn nn nnn nnn ne eS UE ayn SSS SS SgSSSSISISESSSSS SSSR CHEMICAL BEHAVIOR. LL Behavior to alcohol, Alcohol (‘7-1 %) contained in Mayer’s and WNigeli’s solutions, devoid of sugar, changed into acefic acid, ‘Vhe same change was observ- ed in kdji-extract culture. In sake containing 13°3% of alcohol no growth was found, Formation of alcohol and acids. In wort kept at 21-26° C. during to days was formed 4°62 of alcohol. In kGji-extract kept at 19-26° C. in 10 days 2°4% of alcohol and 00589 % of fixed, ‘oco% of volatile acid, while in 15 days °7.5% of alcohol, 0'0392 9 of fixed and ‘01619 of volatile acids, Formation CH, -OF: was formed in culture of k6ji-ex- tract. Assimilation of nitrite, alcohol and glycerine. Assimilates alcohol, gly- cerine, but not nitrite, By these properties this yeast can be distinguished from the preceding varieties, except Mycoderma A, by fermenting saccharose. Further from Mycoderma A, it differs by not assimilating maltose and having various cell forms and also by forming larger quantities of alcohol. Some New Varieties of Mycoderma Yeast. 393 MYCODERMA F (Plate XXVIII F);, FOUND IN SAKE. GENERAL CHARACTERISTICS, Form and usual size. Mostly elongated cells; 175-2 p. b., Cells are rich in vacuoles 2 NOV ple. le containing revoly- ing refractive gra- nules. to the periphery. On sugar-bouillon agar it formsa | sembling to that of Mycoderma yeast B and I. and yellowish white in color. mesenteric film, which alters gradually to yellowish colonies. The liquefaction of gelatine sets in slowly. Giant colonies on wort gelatine are conical in shape P Thin margins are irregularly defined, while the surface are ornament- ed with deep lines regularly running from the centre coarse mesenteric giant colony. or brownish white with a rosy tinge (on wort) re- | On | wort or koji-extract-gelatine, it forms powder like glucose only. Assi- | milates glu- cose, Sac- charose,and maltose. Behavior |Optimum and Growth. to killing sugars. temperature, On the wort or kdji-extract forms a thin white Ferments Optimum temperature 22—24° C. 56° C. for Io m. will cause the death of | the cells, CHEMICAL BEHAVIOR. Behavior to alcohol, Formation Assimilation | of nitrite, Alcohol (*7-19%) contained Mayer’s and Nigeli’s solutions, de- void of sugar, was changed into acetic acid. ‘The same phenomenon was observed in k6ji-extract culture. In sake containing 70°7 % of alcoho/, it grows very well forming a very thick mesenteric film. After 36 days there was found only o-32 % o- 70 ot alcohol. Higher percentage (1373 _%) of alcohol in sake depressed its growth. In k6ji-extract held at 19-26 C. in 10 days, there was found 2°11% of alcohol, 0'0627% of fixed and 0'0057 % of volatile acids, the latter was found to consist chiefly of acetic acid with a trace of dutyric acid, After 15 days alcohol increased to 3°41%, and 0'0219% of fixed and o'0019% of volatile acid were found. | the koji-ex- tract cul- ture. Formation of alcohol and acids. of nlcohol aril CH...OH. PIE AE glycerine. in In wort kept at 21-26° C. after 10 CH,.0OH | _ Assimilates days was found 2°72 of alcohol, | found __ in | all three. It will be seen that, this yeast differs from Mycoderma A, C, E by assimilating nitrite. From Mycoderma B, D by destroying the fixed acids and still from the former by having lower killing and optimum temperatures. Moreover from species D by not showing wedge shape. C and E by growing on sake. From Mycoderma YT. Takahashi. MYCODERMA YEAST I (Plate XXVIII I), ' FOUND IN MOTO MASH. GENERAL CHARACTERISTICS. LL eee ——————— Fos fecal Behavior |Optimum and orm aus usua Growth. to killing este sugars, temperature. Mostly oval or el- On kdji-extract or wort this species forms a finely Ferments Optimum liptic somewhat re- | corrugated film. The color is chalky white in the | only — glu- temperature sembling Sacch. | beginning. It changes gradually into a faint yellow | cose. Assi- | 22 — 24° Cc. exiguus being 1°O- b. and 1°4- The cells 22 22 p. |, at 26°5° €.2 have generally a hemogeneous pro- toplasm _ provided with vocuoles with } I or 2 revolving granules, (on kéji-extract) or bright rosy brown (on wort). The film formation is complet on wort after 2.5 hows On kéji-extract-gelatine it formes a somewhat waxy colonies, but on wort-gelatine rather mealy. The gelatine is liquefied rapidly. Gianticolonies on wort-gelatine have conical form of slight yellowish color. The margin is well defined and the surface is furnished with fine radiating lines. On sugar-bouillon agar it forms rather flat and smooth giant colony. milates glu- COSe, | Sde= charose,and maltose. Ten minutes heating at 5°44 °E.. is not sufficient to kill the cells. Behavior to alcohol. CHEMICAL BEHAVIOR. Formation of alcohol and acids, Alcohol contained in culture solution, devoid of sugar, was con- verted into acetic acid. In sake containing 10°77% of alcohol there After 30 days the percentage of alcohol was found a good growth. was reduced to 450%. Formation In k6ji-extract kept at 19-26° C. after 10 days cultivation were form- ed 341% of alcohol, 0 126% of Jixed and 0'0057% of volatile acids; after 15 days 5:09% of alcohol, 0°0514% of fixed and 0,0361% of volatile acids. In wort kept at 21-26° C., there were found 3°419¢ of alcohol after Io days. CH,.0H was found in k6ji-ex- tract cul- ture, Assimilation of nitrite, alcohol and glycerine, Assimilates all three. This yeast differs from Mycoderma A, C, E by assimilating nitrite. From all the preceding species by its very small size. Further it differs from Mycoderma C and E by assimilating maltose, and from species A, E by not fermenting saccharose. Still further it differs from Mycoderma A, C, E by the growth on Sake, and from species B, D by the faculty of decomposing fixed acid. on wort at 26°5° C. From species F it differs by the very quick formation of the film 1 This is one point which distinguishes this yeast from Mycoderma F, which forms the film after 43 hours under the same condition, Some New Varieties of Mycoderma Yeast. MYCODERMA YEAST M (Plate XXVIII M), ISOLATED FROM KOJI. GENERAL CHARACTERISTICS, 395 Form and usual size Round, oval or long filamental form. ‘The long cells have many vacuoles, in which revolving refracting granules are pre- sent, 2'0-2'5 w. b., and 2515) fb. le Behavior Growth, to sugars. On wort or kOji-extract it forms a rather brownish Ferments and very thick film. On wort-gelatine the colonies | only — glu- are chalky white with central elevation. The margin | cose. Assi- part has very fine lineations. Streak culture on kGji- | extract-gelatine gives light brown colonies. They are rather smooth but the margin part has fine linea- tions. It liquefies gelatine slowly. Giant colonies on wort-gelatine seems white mesenteric and some what moistened, while on sugar-bouillon-agar white with very coarse folds. milates glu- cose, sac- , charose,and maltose. Optimum and killing temperature, Optimum temperature 27 - 30° C. 54°4° C. for 10 m. is not sufficient to kill the cells, it suffices at CHEMICAL BEHAVIOR. Belavior to alcohol. Alcohol (*7-1 %) contained in Mayer’s and Niageli’s solution is changed into acetic acid. is found in the kdji-extract culture. In sake containing 13°3°% of alcohol Formation of alcohol and acids. In k6ji-extract, there was found at 19-26° C, 96 of fixed and 0°00761 % of volatile 2°39.% of alcohol, 0°0374 This fact acids in ro days, while in 15 days was found 2°56°% of alcohol, 00935 no growth was observed, % of fixed and 0'0266% volatile acids. In wort held at 21-26° C. was found 2°11 of alcohol in 10 days. The volatile acid consisted chiefly of acetic and butyric acids. Formation of CH,. OH. Assimilation of nitrite, alcohol and glycerine. ‘Trace was | found in | koji-extract culture. Assimilates glycerine al- cohol, but not nitrite. Thus it is obvious that this yeast differs from Mycoderma Bb, D, F, I by not assimilating nitrite. and from mycoderma A by not fermenting saccharose. from all of the preceding species temperature for the growth. From Mycoderma C and E by assimilating maltose, Further it differs except C by having a higher optimum 390 T. Takahashi. MYCODERMA N (Plate XXVIII N), WAS FOUND IN KOE GENERAL CHARACTERISTICS. Form and usual size. Very long fila- mental form, rich in vacuoles which con- taining revolving Growth. On kdji-extract or wort makes very gzuck develop- ment of the film, which is at first white mesenteric changing afterward yellowish (kOji-extract) or yellowish-brown (on wort). The colonies on wort- Behavior to sugars. Ferments only glu- cose. Assi- milates glu- Optimum and killing temperature, Optimum temperature 23'= (30° aie: The heating granules and the | gelatine seems greyeywhite and mesenteric. Streak | cose, sac- | to 544° C. size: 3 mw. b., 7-| culture on wort or kdji-extract-gelatine gives beauti- | charose,and | for 10 m, 30 p. I. ful greyey coarse mesenteric colonies. It liquefies | maltose. causes the gelatine rather quickly. Gianttcolonies on wort- death of gelatine are white and seem mesenteric without cells, = elevation; on sugar-bouillon-agar it forms a coarse, chalky white mesenteric colonies, CHEMICAL BEHAVIOR. eer Assimilation Behavior to alcohol. Formation of alcohol and acids. of a meee CH.. oH, | 2lcohol an 3° glycerine. Alcohol contained in culture In kdji-extract kept at 19-26° C. Forms | _ Assimilates solution changed to acetic acid. | in 10 days, there wasifound 2°94% | CH,. OH | glycerine, al- The same fact was observed in kGji- extract culture, In sake containing 10°77% of alcohol it forms a yellowish-brown mesenteric film, but the increase of | alcohol at 21-26° C. during 10 days. | the alcoholic content to 13°3%% de- pressed the growth, of alcohol, 0°0864% of fixed and 0047 % of volatile acids, while after 15 days the alcohol decreased to | 2°56%. In wort it forms 39% of | The volatile acids consist of acetic and butyric acids, in kojl-ex- tract cul- ture, cohol but not nitrite. This yeast differs from mycoderma B, D, F and I by not assimilat- ing nitrite, it differs from species C and E by assimilating maltose. species A and E by the incapability of fermenting saccharose. specics M, E, C, A by the growth on sake. From Further from Some New Varieties of Mycoderma Yeast. 397 MYCODERMA YEAST O (Plate- XXVIII O), WAS FOUND IN KOJL GENERAL CHARACTERISTICS. Roem aad usual iar Bebavior Optimum and : Growth, to killing size, = sugars, temperature. Round, clup or On koji-extract or wort forms somewhat greyey | Ferments Optimum long ellipse and the | film with thick foulds. The culture on wort-gelatine | only glu- | temperature latter has vacuoles | gives waxy and more or less mealy colonies carrying | cose, Assi- | 22-24? C. rich in revolving | veins on the margin. | milates glu- | Exposure to | granules, Streak culture on wort-gelatine mesenteric with a | cose, sac- | 544° C. for 2°5-3 MB. b., 10- | slightly greyey color, on kéji-extract-gelatine it | charose,| 10 m.. kills 18 pw. 1. makes a chalky white growth. Giant colonies on | maltose, the cells. wort-gelatine are pasty and half granular, but with | no coloration, on sugar-bouillon-agar they are slightly brown with granular folds on central part | differing from species E, Also, on glycerine solu- tions it shows a most energetic, growth. CHEMICAL BEHAVIOR, . 5 Assimilation Formation of nitrit . = . = } = te, Behavior to alcohol. Formation of alcohol and acids, | of les alcohol and (GET, JOH: fF, glycerine, Alcohol (7-1%) contained in In koji-extract kept at 19-26°C.| Trace of | Assimilates Mayer’s or Nageli’s solution, devoid | for 10 days, there was found 2.339 | CH,. OH | glycerine, of sugar, is converted into acetic | of alcohol, o-o14% of fixed and | was found | and alcohol. acid. ‘The same fact is also observ- | 0'000% of volatile acids, while after | in kGji-ex- ed in k6ji-extract culture. In sake | 15 days cultivation 39% of alcohol, | tract — cul- containing 10°77% of alcohol it | 0°0178%% of fixed and o'o180% of | ture. shows no growth, volatile acids were found, In wort held at 21-26° C. was found 3°41% of alcohol. This yeast is distinguishable from all preceding species by the vigorous growth on solutions containing glycerin (21-26° C.). Like species E the formation of volatile acid is slower than with the other species. From varieties A, E it differs by not fermenting saccharose ; from C and E by the assimilation of maltose. The incapability of the growth on sake distinguishes this yeast from species B, D, F, I and N. The lower optimum temperature distinguishes from species M. 398 Y. Takahashi. MYCODERMA YEAST Q AND R (Plate XXVIII), WERE FOUND IN KOJI. GENERAL CHARACTERISTICS. ST aca e ee) Behavior |Optimum and orm and usua Growth. to killing size, sugars, temperature, Round, ellipse or On kéji-extract or wort both varieties forma very Both spe- Optimum long filament. The | thick mesenteric film of a yellowish-brown color, | cieses fer- | temp, 27-30° revolving granules | The colonies on wort-gelatine are chalky white | ment glu- | C. killing in vacuole occur | and have two main folds on the surface. | cose, sac- | temp. 54°4° very rare, 2-2°5 w. | Streak culture on kdji-extract-gelatine give a very | charose but | C. for 10 m., b., 10-20 p. 1. coarse beautiful greycy (sp. Q) or yellowish (sp. R) | not — mal- mesenteric coating. The former liquefies gelatine | tose. Assi- quicker than the latter species, The same culture | milate three on wort-gelatine gives moistened (sp. R) or dry (sp. | sugars. Q) mesenteric coatings. Giant colonies on wort- gelatine are yellowish with an elevated mesenteric | é ° hd Spe + aa | film, The margin is irregular, Those on sugar- bouillon-agar seem mesenteric and especially in the case of species Q are colored more or less yellowish, CHEMICAL BEHAVIOR. Formation pean Behavior to alcohol. Formation of alcohol and acids. of ] ©, : alcoho! and CH,. OH. ; glycerine. Alcohol (7-1 %) contained in The species Q forms in k6ji-extract | Both form- Assimilate Mayer’s and Nageli’s solution, de- | at rg—26° C. during 10 days 3°829% | ed CHg. glycerine void of sugar, is converted into | of alcohol, -0682% of fixed and | OU in the | and alcohol, acetic acid. In sake containing | ‘o1g99% of volatile acids, while after | culture of 10°779% of alcohol both species | 15 days 475% of alcohol 04869 of | koji-extract. make very quick growth and after | fixed and ‘045% of volatile acid. | Species R 36 days there was found 4°509% (sp. | The species R forms at the same | formed lar- Q) and 5°56%% (sp. R) of alcohol. If alcohol is increased to 13°3% both failed to develope. condition during the first period 267% of alcohol, -o112% of fixed and *o285%% of volatile acids. And after 15 days 3:06% of alcohol, 0561 % of the former acid and :0380 9% of the latter acid. The volatile | acid consist of acetic and formic acids (sp. R) or only acetic acid (sp. Q). This property and the destroying faculty of the species Q for fixed acid | distinguishes from species R. ger quanti- ty. By these properties we can distinguish the two species from all the varieties except A and E, by the fermenting faculty of saccharose, while the former two varieties do not develop on sake and by this properties differ from the species O and R. ~ Some New Varieties of Mycoderma Yeast. 399 MVeCODERMA- YEAST -S- (Plate GXVIH S), FOUND-IN THE AIR: OF AL SARE PACTORY. GENERAL CHARACTERISTICS. Bota andeusdal Behavior Optimum and j rrOW | li shat! Growth, to | _ killing sugars. | temperature, Chiefly round On koji-extract or wort it forms a ring at common Ferments | Optimum and very rare el- | temperature, but on an artificial solution containing | only glu- | temp. 27-30° lipse. The cell con- | alcohol (Nageli’s solution), or glycerin instead of | cose. Assi- | C. Killing tents are very | sugar (Hayduck’s, solution) or Hayduck’s solution | milates glu- temperature bright 2-5-5 w. b., | shows a very energetic growth, It forms on wort- | cose, sac- | lies at 54°4° 3-7 M1. gelatine a waxy conical colony with a ring-like canal | charose but | C. near the top of the colony. Streak culture on wort | not maltose, or kdji-extract gelatine gives a white waxy coating. It liquefies gelatine very slowly. Giant colonies’ on wort-gelatine are a white, elevated mass of waxy | nature. On the central part some petal-like figures | radiate from centre very similar to that of S. Ava/os- porus. On sugar-bouillon-agar it gives rather flat and waxy colonies. CHEMICAL BEHAVIOR. oa ESI ee aS SaaS | . . - Assimilation of nitrite, Formation : Behavior to alcohol, Formation of alcohol and acids, fea of A ea PGE OFT (Fe 3 alcohol. Alcohol contained in artificial In kdji-extract kept at 19-26° C. Forms|_ Assimilates nutrient solution is changed into | for 10 days, it formed 2.619% of | CH,. OH | the latter acetic acid. In sake containing | alcohol, 07028% fixed and o-or2z2 | in kOji-ex- | two. 10°779% of alcohol there was no | % of volatile acids, while after 15 | tract cul- | growth, days 3°24% of alcohol, 003% of | ture. fixed and o:00237% of volatile acids, the latter acid consisting of acetic and butyric acids, In wort | at 21-26° C. for 10 days 3°35% of | alcohol was found. The round form of the cell is the essential character in which this yeast differs from the described varieties. The incapacity of assimilating mal- tose distinguishes it from the preceding varieties, except C and E, while species E ferments saccharose and hence differs from this yeast. The species C has a different cell form and may thus be distinguished from this yeast. 400 T. Takahashi. The properties which distinguish our mycoderma yeast varieties from certain well known mycoderma yeasts : They differ from Mycoderma Cerevisize, vini, Desm, ¥ pe i Sertent: Endoblastoderma amycoides (TSE iV): 2” Endoblastoderma liquefaciens. (These varieties do not cause fermentation). Endoblastoderma glu- comyces (I, II, III, IV). (Do not liquefy gela- tine, ferment g/u- cose, Endoblastoderma pul- verulentum. (Ferment maltose, saccharose), Mycoderma A. by fermenting glucose. Mycoderma B, ditto, Mycoderma C, ditto, By fermenting saccha- rose, and liquefying By not causing fer- mentation of mal- Henneberg’s Mycoder- maa and b, {Ferments, besides glu- cose, levulose and maltose, but not sac charose, Produces acetic ether. The opt. temp. 32—51° C. By not fermenting mal- tose, and the lower tempera- ture, and failure to form acetic ether. and maltose. gelatine. tore, optimum By liquefying gela- | By not fermenting x tine. both, — saccharose Mycoderma |), ditto. 3” By incapacity of fer- menting saccharose and maltose. By not fermenting mal- tose and lower op- timum temperature and not forming acetic ether. Mycoderma 1°, ditto. Mycoderma I’, ditto, Mycoderma I, ditto. By the property of fermenting saccha- rose and liquefying gelatine By not fermentin maltose, co > By liquefying yela- tine. ” Mycoderma M. and N, ditto. Mycoderma ©, and R, ditto. Mycoderma 5, ditto, sy liquefying gelatine and fermenting sac- charose. By liquefying tine, gela- ” EE _————————eeeeeTTTTFTFSeSFSNSSSSSSSSS CO” eer —$—$———— Some New Varieties of Mycoderma Yeast. 401 Although a number of mycoderma yeast varieties has hitherto been isolated by various authors, it is very difficult to ascertain whether our Mycoderma species are identical with any of the organisms described in the literature ; for not only the majorities of these organisms have not exactly been investigated as to their morphological and physiological properties, but also some of these properties are not quite constant. For this reason I shall point out, in the following, only those races which are especially noted by their characteristic properties. Among my cultures, the varieties B. D. F. and I. are most interesting on account of their property of assimilating some xitrogen from nitrite, when at the same time glycerin is applied as the source of carbon. Such kinds of yeast have hitherto not been observed, S. acetethylicus of Beijerinck being only capable of assimilating N from nitrates. The varieties of Mycoderma yeast (D. F. I. QO. R. N.) are so far interest- ing as they can grow in sake containing 10°77 w % or 13°32 w %& of alcohol. Not less interesting is the fact that most varieties of Mycoderma yeast isolated by myself have the property of producing from k6ji-extract besides ethyl alcoho! a noticeable trace of methyl alcohol,’ the latter is frequently, as I have proved, present in common sake. Furthermore all kinds produce acetic acid from alcohol and some (species C.) also from glycerin; certain varieties (O. E. B. C.) form besides acetic acid, butyric acid and a few (E. C.) also formic acid in saccharine solution. 1 This alcohol may be one of the oxidation products of ethyl alcohol. 402 T. Takahashi. EXPLANATION OF PLATE XXIX AND XXX. All of these mycoderma yeast cells were taken from the plate culture of wort-gelatine. Magnified 750 times. f: Fatty matters. m=: Revolving particles. L: Light refracting globules. bl Fig. A. Taken from after 10 days culture of Mycoderma yeast A. Fig. B. The cells of Mycoderma yeast B, after 7 days cultivation. —— Fig. C. * “ 7, erie RON tek ” Fig. D. a 33 i spuds 35/2? ay; ay Figs BE. 55 9 7 =e pialda Jit, aa Ses 9 Fig; F, 95 34 i placket sts. 5,1 Cae es » Hig. i. + ce $5 gpildits: 35 oleae, ” Fig. .N. 53 is 3 poeN, 1 Soy ” Figs @): 55 i 55 eniOy. 3g » Fig... f re J gc fits: |, ones ” Fig. Q. i> 53 > OF. taleees ” Fig. 5. x Pr $5 5 9S: 3" Soe Y OV PONTE pn Se es 9 Sale, a gelatine v - a o ' © Cc v a] ro 5 = v Streak ett TOE AONE tay, te rm ins a ee te a “3 ¥ e . : i ¥ 7 i > ~ is 4 ‘ a ae 7 <— , : - oe A. *y ‘ * ‘a! . ® c. * «ae ot . fh oats'7q *. ae vi WF Law mon © Ona _ mi —_ enaiazeprt ee ® ‘ oo , * ‘ t 7 Sey : o* = fe Ge bak! ‘ ‘ , ; j ‘ A ‘ , ‘ ry ‘ oe f > ‘ he piss ~¢ . , e ‘ “ > " n - ta Aa . . x ; 2 . tad . Pi ‘ . < y , : oe . U 4 ” wr PLATE XXIX. fe ea . 7e~ - ‘= a ia , AGRIC, COLL. VOL. VI. | PLATE XXX. “J == ee we | v P| es eg sh taas! | | sins 5 Ie aitel on-agar. Giant colonies on sugar-bouil Can Nitrite Provide Oxygen in Anaerobic Culture of Bacteria ? BY T. Takahashi. Some time ago Weissenberg reported that Bacillus pyocyaneus can develop under anaerobic conditions when nitrite of sodium is present in the culture solution. It would seem therefore that this microbe could utilize the oxygen of the nitrite when atmospheric oxygen is withheld. Since, however, similar experiments made long ago by Loew with potassium nitrate and aerobic microbes had failed to yield positive results, the result of Weissenberg seemed rather doubtful, for obligate aerobic microbes at least. The writer has therefore made some experiments with nitrite. 0.25 grams of sodium nitrite was dissolved in bouillon and after sterilization infected with Bac. pyocyaneus. The test tubes were kept anerobically (Buchner’s contrivance) at 30-32° C. After 20 days avery thin scum on the surface was observed and the solution exhibited a weak opalescence. A growth worth mentioning had not taken place. Only very few gas bubbles and that only in the first few days had been observed. The control tubes had remained perfectly clear under the same conditions and it seems therefore that the nitrite enabled the growth of a mere trace of microbes. Similar experiments, in all of which the bouillon was rendered weak alkaline, where made with Bac. subtilis, Bac. mesentericus vulgatus, Bac. mesentericus fuscus, Bac. acidi lactici, Hiippe, Proteus mirabilis, and Bac. of typhoid of mice. The result was here essentially the same as before, namely the oxygen of nitrite cannot replace the molecular oxygen of the air in the life of microbes, at least not with the varieties tested. 1 Those culture solutions contained leucin and in some cases sodium acetate as organic nutrients, and were infected chiefly with Bac. fuoresceus liguefactens, Note of O. Loew. a a » 7 : aes my Pye Mier, Be Au ns ‘nia é ve i ; 7 Va ‘ : TViqie “Ss af 9 acl ae tale its 75 joa ies On Manuring with Kainit. S. Suzuki. It has attracted considerable attention that kainit which is not only a most effective and valuable manuring compound but also the cheapest source of potassa in commerce for the agricultural-plants, exerted in certain cases an injurious action, depressing the yield of agricultural crops. This action was ascribed by Detmer to the presence of chlorids in the kainit. The observation of Schulz-Lupitz, however, that the application of lime in conjunction with kainit can prevent the injurious action of kainit! militates against the view of Detmer and renders it very probable that it is the “magnesia content of the crude kainit which causes the injurious action on certain soils.2 Eunenbach®? who made recently some experiments on the action of kainit in water culture, ascribes, like Detmer, the injurious action he observed, to the presence of chlorids, especially of sodium chlorid. How- ‘ever, his experiment can not throw any light on the cause of the injurious action of kainit on certain soils; especially since he carried out his experi- ments in presence of a sufficient amount of lime which of course counteracted the injurious action of the magnesia of the kainit. Thus it can easily be understood why in his case no injurious action of the magnesium salts in the kainit was observed. Schreiber wants to avoid the crude kainit on heavy 1 Many have therefore applied recently kainit in conjunction with Hime, some with excellent results. It is to be regretted, however, that the authors ignored the original magnesia content of the soil. 2 Perhaps only such soils come here into consideration which contain much more magnesia than lime. 3 Landwirt, Jahrbiicher. XXX. Fragiinzungsband HI. 1902, 406 S. Suzuki. soils on account of its chlorid content, while the action on loose sandy soils is beneficial. He also calls attention to the facts that in some cases the magnesia content of the kainit can act injuriously, in other cases, however, very favourably also.!| Such a case was published recently? in which 0.469 of lime in soil had an injurious effect on lupins, and that this was overcome by an application of kainit; in this case it was very probably chiefly the magnesia content of the kainit that counteracted the effect of the excess of lime. This original soil was very probably very poor in magnesia. Gerlach? ascribes the injurious action of .kainit also to chlorids and believes this could be counteracted by lime. But it is quite impossible to conceive here any connection between the injurious action of chlorids and the supposed counteraction by lime. It was very probably the injurious action of the magnesia in the kainit that was counteracted by liming the soil in Gerlach’s case. It is true that chiorids can exert in certain quantities an injurious offect on plants and numerous experiments have been made with sodium chlorid. The results, however, do not closely agree, the doses applied and the nature of the soil influencing the action,4 The depressing action of potassium or sodium chlorid on the starch content of potato’ is probably only due to a certain excess of these chlorids® and does not answer the question whether much smaller quantities could not 1 Central Bl. Agrikultur Chemie, 1897, p. 802. 2 Deutsch. L. Presse, Vol. 23, Nos gt and 92. 3 Deutsche Landw., Presse 1993, No. 109. 4 In regard to the influence of diluted solutions of NaCl upon the crops, Storf, also 7. Kénig in conjunction with Cosack, Boehmer and Weigman have made also investigations (Biedermann’s Central- balt Vol. 13, 1884). These authors, however, applied comparatively large quantities of NaCl and observed an injurious action, A@ig concludes that a water’ containing 1 per mille NaCl should be avoided in irrigation, 5 Sjollema: Bot. Cent. Bl. 1901, No. 33. 6 While with maize a certain amount of NaCl depresses, according to Schimper, the energy of the assimilation of carbon in the leaves, even a 0.5% solution did not depress it essentially with algae (Av Richter, Flora, 1892). On Manuring with Kainit. 407 act beneficially.'. Storp observed that a solution of 0.019% NaCl exerts probably a favourable action on the germination; stronger solutions, how- ever, acted injuriously upon that process. Yarius observed a stimulating 9 action still at 0.49¢.2 Pethybridge® thinks that sodium chlorid exerts an unfavourable action on the development of root hairs; on the other hand he observed with the wheat plant a favourable action, consisting in the production of a deep green color.4 Slomeyer® observed a very favorable action on beans by the application of 100 kilo. NaCl per ha., but he would not infer a general recommendation. It seems to me, however, that it is above all necessary to know the quantity of NaCl already present in the soil and to recommend an application only in those cases where that amount was exceedingly small. Lznenbach® concludes ‘Eine geringe Menge Chlor bekommt allen Pflanzen gut. Buchweizen konnte ich z. B. ohne Chlor gar nicht zur Fruchtreife bringen.”? This agrees very well with the results of Nobbe who infered from his experiments that. 1. Chlorkalium ist die beste Form der Kalizufuhr. 2. Chlor ist nétig fiir den normalen Kreislauf des Buchweizens. t According to Hi/gard the sugar beet can grow well on sandy soil containing besides 0.29% Na,SO, 0.19§ NaCl without a depression of the sugar coutent. ossowrtsch (Journal f. exp. Land- wirtschaft, 1903, p. 44) reports that flax was much damaged by 0.194% NaCl added to the soil. 2 Landw. Versuch stationen, Vol. 32. p. 149 (1886). 3 Bot. Cent. Bl., 1901, No. 33. 4 Wagner observed a very favorable action of small doses of sodium chlorid with carrots, but not with barley (Die Stickstoffdiingung, p. 233). 5 Die Cultur der landw. Nutzpflanzen, I. Bd. p. 330). 6 Landw. Jahrbiicher XXX. Bd. Erginzungsband III., 1902, p. 21. 7 Quite recently Gerneck (Uber die Bedeutung der anorganischen Salze fir die Entwickelung und den Bau der hdheren Pflanzen) made a series of experiments and concluded: “ Dre Kulturversche mit Kresse Naben also ebenso wie die Weizenversuche die Angaben von Lesage tiber die Vermehrung des Palissadenparenchyms durch Kochsalz bestatigt, dagegen nicht seine Angabe, Kochsa!z bedinge eine Abnahme des Chlorophyllgehaltes. Meine Maiskulturen ergaben in erster Linie das Resultat, dass Mais sehr wohl 0.5 proc. NaCl vertragt und in dieser Lésung sogar zur Fruktifikation gelangen kann, entgegen den Angaben Schimpers, dessen Maispflanzen in 0.5 proc. NaCl nicht tiber die Anfangsstadien der Entwicklung hinauskamen, Eine Concentration des Kochsalzes von 1 proc, konnten meine Maisexemplare nicht vertragen, trotzdem ich ihnen allen irgend méglichen Schutz angedeihen liess.” 408 S. Suzuki. Adolf Mayer also admits that potassium chlorid is the most favourable form of potassium ; it accelerates the ripening of the fruits, by supporting the transportation of carbohydrates from the leaves to the fruits. He ascribes on the other hand the poor development of the buckwheat in absence of chlorids in Modbde's experiments partly to the abundance of nitrates.2 H/dstermann has studied the action of sodium chlorid upon some Graminece. The solutions contained from oos5 to 5% NaCl. He observed that sodium chlorid has already in a small quantity (0.19% NaCl) a depressing influence upon the transpiration, and that the energy of assimila- tion of carbon decreases already ir solutions of 0.059 NaCl, and at 1% concentration it seems to prevent the assimilation altogether. When the NaCl content is so small that it can not cause any damage, the plant will assume gradually the character of a xerophyte, the walls of the epidermis cells become thicker and the mass of the vascular bundles becomes larger, the number and size of the stomata become smaller and the hair formation was increased. Thus far no explanation in regard to the physiological action of sodium chlorid in plants has been given, but it has been observed by Chittenden that the vegetable diastase can act more energetically in presence of a small quantity of sodium chlorid (0.24% NaCl) and similar observations have been made recently in regard to the animal diastase by Wachsmann.* A. Mayer* found that 1% potassium chlorid retarded the diastatic action, while smaller quantities exerted no decisive effect on the result. A favourable action of sodium chlorid in the living plant might casily find explanation by the observations of Wachsmann and of Chittenden, since the transport of starch from the leaves to the growing tips might be facilitated by an increase of the diastatic action. All experience tends to show that favourable effects will result as long as the absolute quantity of sodium chlorid remains below a certain limit and 1 Journal fiir Landwirtschaft. Vol, 49, p. 42. * A recent writer recommends a dose of 20 kilo. NaC! per ha as very favourable for all kinds of market garden crops. * Pfligers Archiv, 1902, Vol, 91, p. 191. * Journal fiir Landwirtschaft. Vol, 49. p. 57. On Manuring with Kainit. 409 that with the rise above that limit injurious effects will appear and increase. If this is so, then it is above all absolutely necessary to determine ana- lytically the original chlorine content in the soil before the question of application of sodium chlorid on this soil can be taken into consideration, and even then the results might differ according to humidity or dryness of the climate. In order to observe the limits between the beneficial and the injurious influence of NaCl upon certain plants I have made several experiments with a soil that had not been manured for 4 years and had been much exhausted by various crops. ‘This soil contained in 100 parts (air-dry) only 0.0055 gram NaCl or 0.0033 gram Cl. I. Experiment with pea. 3 Wagner’s porcelain pots, each containing 6.7 kilo air-dry soil, served for the experiment. As general manure for each pot was applied: Double superphosphate .......-....... 12.0 grams S00 MU a5 92: el sie Peon Sulplate ol... ..-..s.. eo See PGtassiumm Garponate! .,.....2........+ 10.0 5 While one pot (A.) served as check, the other two (B. and C.) received O.I gram. and 5.0 grams. NaCl respectively. 15 seeds of pea were sown in each pot on December 1ogth (1902), and the young shoots reduced to 6 of equal size four weeks later. There was no decided difference in hight during the vegetation. The flowering started on April oth and had ended on the 1st of May. Uptothe flowering period the pots received almost every day 300.0 c.c. water, later on 500.0 c.c. The plants were cut on June 2nd and weighed in the air-dry state with the following results : 1 Potassium carbonate was applied 2 days later. 2 Hence one kilo air-dry soil received 0.015 gram. NaCl in B, and 0.75 gram in C., and the pencentage of NaCl in each pot was : A, B. 4 0,0055% NaCl, (original), 0.007 % 0.0805 % Aio S. Suzuki. Total weight. Weight of seeds. A. (Control), }15.0 grams, 60.0 grams, B. (0.1 gram. NaCl). TSO Wes OW5 Bs (5 (ele 55 S P2A4'Ouees 62%160) &: 2. Experiment with buckwheat. 4 Wagner’s pots containing 5.4 kilo. air-dry soil received each the following manure : Double superphosphate... 9.6 grams Sodium nitrate: sce Aro ey; Ammonium sulphate ...... Ow Potassium carbonate .. ... 8.0 ,, (separately applied). One pot A. received 0.015 gram. NaCl, another B. 0.148 gram. and the third C. 0.74 gram sodium chlorid for each kilo air-dry soil, while pot D. served as control.t 35 seeds of buckwheat were sewn in each pot on March 28th (1903). The young shoots were reduced, to 6 of equal height on April 27th. Length measurement was made on April 30th, with the following results: Average length of the stalks, A, (0.015 gram NaCl for each kilo soil). B2°5 1Cins b, (0.148 ” ” ” ” ” ” )): 51.3 ” C, (o 74 ” ” ” ” ” ” }s ' 45.8 ” D, (Control), fons) 5, Although the plants in pot C. were shortest, they had the thickest stems and most branches. On July toth, the crop was harvested and left to become air-dry. The weight was as follows: 1 Therefore the percent amount of NaCl in each pot became as follows : : B, OF D. (original soil), 0,007 % 0.0203 % 0.0795.% 0,005 5 On Manuring with Kainit. A4Il Total weight. Weight seeds. = A. 0.9070% NaCl. 118.9 grams, 49.4 grams, B: 0:0203% ,, 13220 9 fee CM O:0705)9%. 5s A372 Sas Sao ID SOGOES'S T2265, 49.0 3. Experiment with rice. 4 Wagner’s pots were filled with 6.2 kilo. air-dry soil taken from the same unmanured field as served for the preceding experiments. As general manure for each pot served: Ammonium sulphate ........ sie Se a 4.25 grams. SGU THikaverss i... s+... eS ae Fe Double superphosphate...... Api ee hy 8) ” eorassiuni SHIpMace. 0... el. eee 8.5 * while the amount of NaCl applied was as follows : (POI UAC Coe ae a eer 6.4 grams NaCl! 0 Ae oo a ere 9:6 \) is ON CR ne TOMO: 55 oi ~) 12, (CR ferns ol) no addition On July 30th (1903), the young rice plants from the seedbed were trans- planted into the pots, each receiving 3 bundles of three healthy individuals of equal size (about 42 c.m. long). Towards the end of August a decided difference in plant growth was noticed in favour of the control pot. The plants that received NaCl were more or less injured. Moreover it was very noticeable that the plants in C which had received most NaCl had the deepest green leaves, although the height was far behind that of the other plants. On September 25th all 1 Hence the percent content of NaCl became as follows : A. B. iG: D. (original soil). 0.1087.% 0.1603% 0.263596 0.00556 te 412 S. Suzuki. plants were in flower, and on November 2nd, the crop was harvested and left to become air-dry. ! The weight was as follows: eT | Number | Weight of | Weight of Weight Total of stalks | fall grains empty of i ht bearing | (unhusked), grains. straw. pen ears, gram. gram, gram. gram: } | | A, (6.4 grams. NaCl). | 49 52.5 3.2 59.5 WLS -2 B(6:605 5; ab): | 55 43.4 1.8 59.0 104.2 Ga (6) +) sy stab): | 33 11.6 2.2 41.3 55-1 D. (Control), | 54 | 64.2 2.8 70.0 137-7 These experiments with pea and buckwheat show that moderate quantities of NaCl can exert only a beneficial action, while with the in- crease of NaCl in the experiment with rice in contrary a depression resulted. [t is therefore absolutely necessary to take care of the amount of chlorid applied and to consider the original chlorine content of the soil. Further it must be mentioned in regard to the experiment with rice that the depression would not have been so great in the open field, since the rains and irrigation water would have washed out a considerable portion of the salt. Now let us see whether the kainit could act injuriously on account of its chlorine content. Since the usual doses of kainit do not exceed 800 kilo. per ha,? the chlorine content of this amount would certainly be insufficient to damage the above crops on soils containing but traces of chlorids. Let us make a calculation on this point. We obtained the following numbers : 1 At this time the leaves of most plants had turned yellowish, but the plants in pot C, still showed a green color, ‘This agrees with the observation of Pethydridge mentioned above, that chlorids produce a deep green color, 2 Maercker mentions, however, certain cases in which the enormous dose of 2,000 kilo kainit per ha had been applied. In such a case the possibility of some injurious influence of the chlorids present on certain crops as potatoes could not be denied. On Manuring with Kaimit. 413 Sodium chlorid was applied in the experiment with NaCl kilo per ha, Reet Sete 505 peak aces dee ee eee 1018.2 Eyxekwiheab 4: <..2.sh0..0+ a ER 1051.8 LSC <=, 2) aa eee Nese dee Ba 2 1682.9 These amounts of NaCl correspond in regard to the chlorine content to kainit,! kilo per ha: 1: INRA oh cd ao en ce 4,402 LUCA lelecie 2 (ACM Ue AER aces Boe ns ee 4,547 ECS Eye EM Satins xk Con aiep eee aescce s+ 7,259 We find from these numbers that the chlorine content of even 4,400 kilo. kainit per ha would not be sufficient to yield a depression of the yield with buckwheat or pea, while only the immense dose of 7,259 kilo kainit per ha would lead to a depression with rice. But such an enormous quantity of kainit is excluded from practice altogether. My further ex- periments with kainit described further below in which this was applied in pot experiments, the doses corresponded to a ratio of 4,980 kilo. kainit per ha, but even at this ratio no depression with rice and pea resulted. In those experiments to be described presently the kainit was applied in following proportion : Kainit, : Per pot Per ha grams. kilo. inGlewiheat® tc sosgene Sthae a 8 GS 35592.0 PEVAISC GOUGET Ge ta cement eles Lasers 12.22. = 2,488.0 OC Ae ee 2 24.44 = 4,977.0 Peak Ain un heRac emai d Sioa. cacy 24.44 = 4,977.0 1 yoo parts of our air-dry purified kainit (analysed by myselt) contained, (CEO Ne sree eT re oO BO ae ce tena vot oteet 17.67 DEO! Gs ied. sous nel eae 8.85 Chlorine, as NaCl ......23.125 Buckwheat was cultivated in a smaller Wagner's pot the surface area of which was = sgs4ygqg ba. i For these three crops served ‘larger pots’ the surface of which was = sgy'gq ha. (eo 414 S. Suzuki. The following experiments were made with the view to decide, whether the hainuit could act injuriously on account of its magnesium content. Such an injury would have been expected, however, only on soils which are considerably richer in magnesia than in lime. The results of the writer show that a moderate surplus of magnesia over lime would not yet interfer with the beneficial action of the kainit, since the magnesia content of the quantities of kainit applied per ha is not so large to yield an undue increase of the total magnesia in the soil. Of course there will be some differences noticed when the kainit is applied in the autumn or in the spring. 1. Experiment with Buckwheat. 4 Wagner’s pots, each containing 5.4 kilo air-dry soil received as manure, each: SOCNin PNOSMMAte. cca wen ante ace ane ne 16.0 grams. Ammoniund Sulphate. sa, scene cs-e tad An on PIO CREME MGs te py eer escent wee te ALO” Besides, the following compounds were applied : Pot A..,.Potasstum sulphate 72... .7:: 3.2 grams. DB Pieei nite agree eotclee: +, Giee 0578 iia, {Potassium sulphate: ..s...22, 3.2 A et | Magnesium Sulphate: yok 5.78 3 Taio ees ya iahy dks Fat Oates 54 : eller ey eo eee Oi0G2 es 20 seeds of buckwheat were sown on March 28th (1903) and the young shoots reduced to 6 of equal size on April 27th. Length measurements were made on April 30th and May 17th with the following results : 1 ‘The chlorid content (as NaCl) of this quantity of kainit was 2.262 grams. NaCl. Hence the percentage of NaCl in the soil became as follows : Porte Coys fone eihreneaeiees ety ts 0.0055 % NaCl (original soil) tafdhl 8 Wo be Pet eR cc cere 0.0529% ,, 2 ‘This amount of CaO is equivalent to that of MgO contained in the kainit, On Mannring with Kainit. 415 a Average length of the stalks, April 30th. May 17th. A, (KSO,). 54.0 c.m. | 81.4 C.m. B. (Kainit). | 485 | 78.9 » Ge (K,SO,+MgSO,). ROM os 79:9. 9s D. — (Kainit + CaO). 50.0 4, | 78.6 The crops were harvested on July 1oth and left to become air-dry. The weight was as follows : Total weight. Weight of seeds. A. (K,SO,). 123.6 grams. | 54.8 grams. B. (Kainit), 13217 = | 56.1 Cc. (K,SO, +MgSO,)- ISRO] 53 | 57.0 D. — (Kainit + CaO). 7 ae ae 2. Experiment with Phaseolus. 4 Wagner's pots each containing 6.7 kilo air-dry soil served for the experiment. General and special manures were applied in the same pro- portion (per kilo soil) as in the experiment with buckwheat. I5 seeds were sown on March 26th and the young shoots reduced to 4 of equal height on April 21st. The plants had developed very well, starting the flowering on May 17th, but there was no decided difference in height during the vegetation. The plants were cut on July 20th and weighed in the fresh with the following results ; 416 S. Suzuki. EERE eR Weight of fruits. Weight of seeds, A. WEEASO,): 60.5 grams, 43-5 grams, B. (Kainit). | 66.7 o 50.1 5 (Ge (K,SO, + MgSO,). 70.9 - 49.8 - D. (Kainit + CaO). | 63.0 48.5 AA These two experiments show that the action of kainit on our soil did not differ much from that of the artificial mixture of potassium and magne- sium sulphate, but the addition of CaO did here not promote the action of kainit. Kainit can very probably act injuriously only on such soils as contain already a considerable excess of magnesia over lime; it would increase the injurious excess of magnesia still more. Only in these cases a simultaneous application of lime with the kainit would be in order and act beneficially. Two more experiments with rice and pea were made: 3. Laxperiments with rice. x 4 Wagner’s porcelain pots were filled with 8.2 kilo. air-dry soil. As general manure for each pot served : POCMUMpPhOspiis te tw «Selene wasn ann eat 20.0 grams. Ammoniumsulphate ........... ree £:0n Ee $50) CLELIENITNT OL CCOe Ae ees ia te tas anes ukneee BO) ae and in order to increase! the magnesia content of the soil 83.2 grams. finely powdered magnesite? (for each pot) were applied. The amount of com- pounds applied were as follows : 1 ‘hus, the ratio of lime to magnesia in the soil became ae (The analytical data for this calculation will follow below. * Magnesite was imported from Germany and contained only minute quantities of impurities, On Mannring with Kainit. 417 Pot A..:. Potassium sulphate ...:.:s%- 8.0 grams. Bipaleainit: sc. eee eee kee SA Potassium sulphate......... 8.0 ze ee sulphates. «..- 144t) , LS | Sins Spies 2 as 7s PAAAS 5, ‘aes Ee Tee re eee m2 2.304 .,, The transplanting of the young rice plants into the pots (each receiving 3 bundles of 3 individuals) took place on June 24 (1903). Although this experiment was carried out in a glass house, the treatment was the same as in the field. The flowering started at the beginning of September. The plants were cut on November 2 and weighed in the air-dry state. The result was a follows: Number Weight, grams, of stalks bearing ) = Se fe a ears. Total eae Full grains Empty | (unhusked).| —_ grains, A. (K,SO,). 56 190.8 108.2 73.4 4.3 Be (Kainit), 63 200.4 119.4 $6.6 3.4 C. (K,SO,+MgSO,). 58 200.1 | m28 | 845 2.9 (Kainit4+ CaO). 67 192.2 106.7 | $1.6 3-9 Also this experiment did not yield results unfavourable for kainit ; in contrary, the kainit acted better than the equivalent amount of potassium sulphate. 4g. Experiment with pea. In this experiment more powdered magnesite was added than in the former case and the ratio of lime to magnesia was now changed in 8 pots to reach 4. The general plan of this experiment will be seen from the oe ~ following table :— 418 S. Suzuki. : g 3 3 |Z a g aie 3 E Number of pots. E & ener a ss 2 |£| ¢ : e a3, Om ae a] + O & = 8 3 = a Nn © O GO) a) x 3S oP 18 cS ise eb) ea 2) = Ss) 7, = Ss 1 7.8 kilo. | 106.3 g.| vo = = ee) 34. 4a oe eee aes a LEI | Sa eanOLp DESE 9°90 Il. ” ” a pet me ” a5 aaa: a III : SP ANegie )) — same ej K,SO,+NaCl...... ihe 2 te ” ” oe “a ” —— _ —_— a ae Kainit+ CaO es : 24 43 2 a aE ia 2.364 &. = AW a wrene nm oc r oeH4 Vie “5 “5 oo 5 = = ” —— ” a = tnd ee K,SO,+ ome » » he £5 8.0 g — 14.44g¢.| — 5.7.2 ~ MgSO,+NaCl Zev g9U, + VITI. : ain r = ‘ Re a On December gth (1603), 15 seeds of pea were sown in each pot and the young shoots reduced to 8 of equal height on January 28th (1904). The flowering commenced on April 8th and had ended on May 3rd. The treat- ment (watering, etc.) was not essentially different from that of the first experiment with pea described above. The crops were harvested on May 3rd and weighed in an air-dry state with the following result :— 1 According to Katayama’s analysis (see Bulletin, College of Agriculture, Imperial University, Vol. VI. No, 2. p. 104) our soil from Komaba contains 0.55% CaO and 0.45% MgO. 2 NaCl was added here in the some ratio as in the kainit, for check. On Manaring with Kainit. 419 —————————— Total | Weight | Weight |x ne.| Weight | Weight |.) werehe. of total of one : of | of total | of one |* umber ss fruits, | fruit, .. | seeds, seed, a: aces grams, | grams, fruits. | grams, | grams. | seed. We 85.0 5a 1.1 49 | 45.6 0.220 | 205 PPE See ease kos ce ++ | 2: 80.0 [ye 0.97 | me || ABA 0.211 204 average, 82.5 52.5 1.04 | 51 44.4 0.216 205 | : = 7 3- 61.2 45-9 | 1.15 | ‘40 37-9 | 0.217 | 174 ey NACL oe. es se | 4. 64.9 46.3 27, 37 37.8 0.223 169 average. 63.1 46.1 1.21 39 37-9 0.220 172 5 76.7 49-7 1.15 43 41.9 0.212 198 eseitmantnt CAO! 25 - cs awetens | 6. Gscif 48.2 1.07 45 40.5 0.216 189 bees 76.2 49.0 Tene 44 412 0.214 194 ie 69.5 44.1 iene 39 36.6 0.239 153 K,SO, + MgSO, + NaCl 8. 79.4 48.8 | 1.25 39 39.2 0.233 168 average, 74.5 46.5 1.19 39 37-9 0.236 161 In all our experiments therefore kainzt has always acted very favorably and no case was observed in which the chlorine content or the magnesia content would have interfered with the production. Therefore I am inclined to believe that cases of a depression by kainit can only be restricted to soils which contain quite an undue percentage of chlorids or of magnesia. Only in the latter case, however, a simultaneous application of lime would counteract the depressing effect of kainit. —_——_—_—> 0 —__—__ On the Influence of Various Ratios of Phosphoric Acid to Nitrogen on the Growth of Barley. Rana Bahadur FROM WE PAT: Numerous investigations have shown that according to different manur- ing, the ratio of N: P,O, in the plant changes considerably. According to Atterberg the oat grain showed a ratio of N: P,O;=100:50 to 55., when nitrogen and phosphoric acid are present in the soil in a favorable ratio. Stahl-Schreder,' however, could not always observe this ratio. With an excess of phosphoric manure only, the ratio 100: 52 was observed. Soils not rich in phosphoric acid gave in one case the ratio 100: 34; in others 100: 20 to 30. This author concludes that for the grains from his soil the proper ratio was 100:35 to 4o. If more phosphoric acid was found in the grains, it would indicate need of nitrogen in the soil or excess of phosphoric acid. When, in the contrary, the amount of phosphoric acid is smaller, it would indicate a need of phosphoric acid or an excess of nitrogen in the soil. Atterberg observed for oats the following limits : anne ener eee eee eee rere Grains. Straw. 1 Journ, f, Landw. Vol. 52, p. So. 422 Rana Bahadur. He believes that the ratio of nitrogen to phosphoric acid can vary between the extremes 100: 15 and 100: 83. Now the question seemed to me of some special interest, whether on the loamy humus soil of our college farm, manuring with nitrogen in dif- ferent forms and in certain ratios to phosphoric acid, would yield also the same ratio in the grains, and furthur to observe which was the best ratio of N: P,O, in the manure for barley on our soil. It was probable from the outset that nitrogen in form of ammonia would not yield quite the same result as the nitrogen in the form of nitrates. According to Wagner one gram ammoniacal nitrogen can produce in average 56.97 grams oat straw and 41.33 grams oat grains; while one gram nitrate nitrogen 58.03 grams oat straw and 42.53 grams oat grains, under otherwise favorable conditions. In applying nitrogen in the form of ammonium salts, it was furthur to be considered on the one hand the nitrification of it in the soil, and on the other hand that a certain excess of ammonium salts can act injuriously. Prof. Aso of this college has recently shown also this injurious property for rice and he kindly permitted me to make use of his data for this article :— Three pots, each containing § kilo soil which contained 119% of humus of weak acid reaction were manured as follows per kilo: A B. C K,O. 0.8 gramasK,SO,. | 08 gramas K,CO,. 0.8 gram as K,SO,. Pee anes 1.0 grams as Double 2.0 grams as Double al mV as Nz (13 Jee ‘ $ P,0;. f.o.gramas Na,TAr Us Superphosphate. Superphosphate. (NIT,).SO, 2.5 grams. 2.5 grams, 5.0 grams, Three bundles of rice shoots, each bundle consisting of three individuals, were planted in each pot July 13. The difference in growth became soon very marked, and the injury by an excess of ammonium sulphate! is also 1 The total quantity of ammonium sulphate in pot C was as much as 40 g. i On the Influence of Various Ratios of Phosphoric Acid to Nitrogen. 423 plainly shown by the plate No. XXXIII. The harvest on November 6 yielded the following numbers. A. B. Cc. grams. grams. yrams, Total harvest (air dry) ......... 279.0 337-5 35.0 SURI agti-hn cpa gS CoCo EEE RE REE 151.0 195.0 9.0 RM PANE eo. -< = 55.0. 5 REP DESOETO! NS Sis dongs vecncs 54.0 5.4 4.2 <0 On the Application of Freezing in the Preparation of Certain Articles of Food in Japan, BY T. Katayama. There are, in Japan, prepared three commercial food articles by the application of frost and subsequent drying. Since this procedure is not practiced in other countries as far as I was informed, some observations in this line may be welcome. The three articles in questions are: KGri- Konnyaku, Kori-Tofu and Kodri-Mochi.! I. Kori-Konnyaku. Kori-Konnyaku is made from tha powdered root of the Konnyaku plant (Amorphophallus Rivieri or Conophallus Konnyaku) which consists to the greater part of mannan.?. The common Konnyaku is prepared by boiling the powdered root with some lime water which destroys the sharp taste of the crude root. The product is formed in tables resembling in its con- sistency gelatinized agar or stiff starch paste. This product contains a high percentage of water, and is, therefore at summer time easily attacked by bacteria. When the gelatinous masses are subjected to freezing, a large quantity of water is forced out chiefly in form of ice and one can easily observe that after thawing up the physical state has completely changed, a very porous 1 Kori is the Japanese word for ice. 2 This was discovered in our laboratory by C, Tsuji; Cf, these Bulletins Vol, IL, No, 2. 434 T. Katayama. condition having been created, the elasticity having decreased, also the thickness; while the feeling to the touch is no longer smooth but rather rough. For my trial served a piece of Konnyaku of tabular shape 6.8 c.m. in width 1.2 c.m. thickness and 15.4 c.m. in length and, weighing 200.5 grams. The loss of water by the first freezing during exposure in a cold night of January amounted to 86 grams. After three consecutive freezing operations, the weight was reduced to 38.6 grams. It was now placed in an incubator at 18°C., where the weight diminished in two days to 9.8 grams. After three days more, the weight was 7.4 grams. There was no growth of microbes noticed.’ The piece was now full of minute pores and measured 5.5 c.m. in width 0.5 ¢.m. in thickness and 11 c.m. in length. The control piece lost at room temperature daily 4-7 grams of water by evaporation. When it was placed in the incubator at 18°C. for 2 days the piece weighed still 125.1 grams and was covered with numerous colonies of bacteria and mould fungi. After 3 days more the piece weighed still 37.1 crams, was hard and nornlike on the surface but still very soft in the interior. No pores were present. The action of the freezing process becomes evident at once when two such pieces are compared. UW. Kovi-Tofu. The tofu consists of the principal protein of the soy bean! and is pre- pared by mixing the hot aqueous extract with liquids containing calcium and magnesium salts ; hereby the soluble alkali phosphates holding the pro- tein in solution are decomposed and the protein, probably also combined with some lime or magnesia, is precipitated, It is then brought into tabular form.2 It contains like the Konnyaku a very large amount of water (ca. 1 According to Ostorne (Journ, Amer, Chem, Soc., vol, 20, p. 419) the chief protein constituent of the soy bean is ¢/yveiuin, a globulin similar in properties to legumin, but containing more sulphur and 0.5 per cent, less nitrogen than this. —— 2 Cf, These Bulletins, vol. If, No, 4. On the Application of Freezing in the Preparation of Certain Articles, 43 oa | 89%) and thus being very liable to putrefaction, it has to be prepared fresh- ly every day. A durable form is prepared by freezing during the winter, and is called then Kori-Tofu, an air dry, very porous and light product. For my observation served half a piece of fresh Tofu of tabular form, 25 c.m. in width, 2.4 c.m. in thickness and 12.7 ¢.m. in length, weighing 119.0 grams. It was exposed to the air in January at—5°C. In the following morning, numerous ice needles were found piercing through the whole piece and water more or less frozen had been secreted. After thawing up and wiping dry the surface with some filter paper, the weight of the piece was now only 76.5 grams. On repeating that process ‘the piece weighed now only 43 grams. This piece was kept now in the incubator at 18°C., whereby the porous mass diminished in weight rapidly, and became hard but remained porous. After 5 days the weight was 15.2 grams and no trace of fungi and microbes had developed on the surface. The control piece lost at room temperature every day only 4 grams of water by evaporation, and when placed after 2 days in the incubator at 18°C., it acquired on the one hand soon a putrid odor and on the other it commenced on the edges tc become hornlike and very compact and hard. After 5 days, it had only decreased to 25.3 grams Now, ifthe Kori-Tofu is compared with this dricd Tofu, a very great difference in the volume is noticed. The Kori-Tofu which had numerous pores by the formation of ice needles in the interior shows now a larger volume inspite of its containing less water than the control piece of unfrozen Tofu. The beneficial action of the freezing process is evident at a glance. The drying can be accomplished quickly without any putrefaction and the digestibility of the product is insured by the great porosity by which it resembles baked bread. WI. Kéri-Meent. Mochi is prepared from glutenous rice and has a consistency of thick starch paste. The durable K6ri-MGchi is prepared from this by the freezing process. The frozen product must not be allowed to become 436 T. Katayama. too warm in drying it, since the pasty character would be more or less reproduced. ! It is kept in the shade at low temperature exposed to the wind whereby a very porous snow white brittle mass results. A fresh tablet of 25 grams in weight was after thorough freezing kept at 20°C., whereby it decreased to 15.2 grams in 3 days. The control piece weighed at this time 17.8 grams, but it was horny and very hard at the surface and on the edges. Such a condition renders the product very difficult to digest. Finally it may be mentioned that also the so-called Kanten, known in other parts of the world as agar-agar, which consists of galactan and is prepared from marine algz is in the fresh state subjected to the freezing process, whereby it assumes the well known highly porous condition. Since this galactan. as well as the mannan above mentioned, serve as an article of food in Japan, it is very probable that there exist in the human intestines galactase and mannase which transform these polysaccharides into the respective sugars. CONCLUSION. The freezing process of articles of food, rich in water, for the purpose of subsequent drying renders the products highly porous which condition is not only essential for digestion but makes possible such a rapid drying that changes by microbes and fungi are excluded. 1 Glutinous rice contains dextrin besides starch, It is well known that starch paste loses its past- ing properties completely by freezing. Or + Ee Note on the Detection and Determination of Fusel Oil, BY T. Takahashi. The quantitative determination of the fusel oil leaves still much to be desired, since its quantitative composition varics to some extent. Asa comparatively satisfactory method is considered that of Xdse. Recently, af, Komorowshy? has suggested a new method, which is based upon the red color Poduced by salicyl-aldehyd (ortho-oxybenzaldehyd) and sulphuric acid. . The writer has substituted for the salicylaldehyd benzaldehyd and various other_aldehyds in this reaction in form of a 1-2%% alcoholic solution and tested commercial fusel oil, as well as pure isoamylalcohol, isobuty!- alcohol, isopropylalcohol and common propylalalcohol. A satisfactory reaction for fusel oil and these alcohols however, was obtained only by. application. of benzaldehyd, anisaldehyd, and vanillin.? It was of course necessary to compare their behavior to SO,H, alone, in absence of fusel oil, to the reaction in the presence of fusel oil. After many tests and com- parisons which may be here omitted, the writer recommends the following way: : To 4-6 c.c. of the fluid to be tested in the test tube add 5-10 drops of 1 Chemikerzeitung, Nr. 88 and “Die deutsche Essigindustrie "? 1903, VII. Jabrg. Nr. 49, S. 385. 2 Fromaldehyd, propylaldehyd, thio-oxybenzaldchyd and chloral gave unsatisfactory reactions. x 3 Komorowsky’s test with ortho-oxybenzaldehyd requires especial care, since this aldehyd pro- 438 T. Takahashi. the color appearing: If fusel oil is present, the following colorations will appear: ; With benzaldehyd :—Redish color above a yellow layer. With anisaldehyd :—Brownish yellow, below green, and yellow layers, the former altering to red, the green to blue and the yellow to brownish- purple and violet. When after some time the fluid is shaken, it shows a purplish red color. With ortho-oxybenzaldehyd :—Purplish layer above a red layer. With vanillin the method is a little different. About 5 c.c. of a 196 alcoholic solution of vanillin is added to an equal volume of concentrated SDH ie and well shaken. After the generation of greenish yellow coloration an equal volume of the testing fluid is added. If the fluid contains fusel oil, the color turns to blue. For a tolerably satisfactory quantitative colorimeric estimation, I propose to proceed as follows : Take roc.c. of the testing fluid and fusel oil of known strength contain- ed in 189J alcohol and pour each into a small cylinder. Then add well mixing 2 c.c. of 199 alcoholic solution of anisaldehyd and next carefully add 20 c.c. of strong SO,H,, and compare the purplish tinges produced by the fluids. Fusel oil, Fusel oil. Fusel oil. Fusel oil. “0001 %. 7001 %. 7O1%. EG: 2 ee ea ae ee = er Peach color aft. 30 S. Peach colored layer Peach and below Yellowish layer after 10 Peach violet aft, 6 m. | aft. 20S. Vivlet below | yellow colored layer | S. Red and below yellow Faint peach colour | yellow layer aft. 3 m. | after 205. The former | lay after 30S, Purplish, when shaken afterrom. | A yel/owish each | alters to violet after | below crimson and yellow _ color when shaken | 1 m., purplish-red | layers after 1m, Crimson | after 10 m. | after3m. Vzo/et when | ved after shaken in ro m, shaken aft. 10 m. - Pa ” ‘ a *" s . ae ’ c + hac ‘ " . > = at . is ’ ‘ aly + \ ay @ = i = Va aw y . ee : ays d a > ry b i. ? 7 =! 7 C@« ° f «7 q 4 i " - na a din a > Pa _ ‘ A? fe : j A _ RAIS ay a — ‘ Ss a "t ~ — S TokyS Daigaku. Nodgakubu fe, Bulletin T64 v.5-6 Biological & Medical Serials PLEASE DO NOT REMOVE CARDS OR SLIPS FROM THIS POCKET a UNIVERSITY OF TORONTO LIBRARY kl ee es ee ee eed eee eet eseeet okapgsty hs OEP OE Ths EP Ed PEPER MRED Cette SB ae STAR REE ef po ssi gph droccces ' ve bs * i . ° ’ " i J ” . » > a % ’ ‘ w , A ee tps cae st ins t “>