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Full text of "Bulletin"

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THE 

BULLETIN 



OF THE 



COLLEGE OF AGRICULTURE, 



Tokyo Imperial UxivERsrrv 
JAPAN. 

Vol. V.—- o 
1902 — 1903. 



Digitized by the Internet Archive 

in 2^©^h funding from 

IJnR/ei%ity of Toronto 



C 
o 



V. 5-6 



http://www.archive.org/details/bulletin5v6toky 



COXTKX'IS OF X'OLl'Ml': \' 



1). KITAO : Inwiefern kann man das Holz als einen isotropen Korper 

betrachten ?---------... i 

T. IKEDA : Studies in the Physiological Functions of Antipodals and Related 

Phenomena of Fertilization in T.iliaceae. I. Trycirtis Hirta. - - - 41 
K. TOYAMA : Contributions to the Study of Silk-Worms. I. On the Embryo- 
logy of the Silk-Worms. - - - - ~ - - - - 73 

N. NITTA : Ueber das wirksame Princip des Tuberculinum Kochii. - - 119 
O. LOEVV und Y. KOZAI : Ueber Ernahrungsverhaltnisse beim Bacillus 

Prodigiosus. - - - - - - - -- - -137 

]\I. TOYONAGA ; Ueber die Vertheilung dcs Kalks in Thierischen Organ- 
ism us. - - - - - - - - - - - -143 

S. SAWAMURA : On the Digestive Porer of the Intestinal Canal. - - 155 
O. LOEW and S. SAWA : On the Action of Manganese Compounds on 

Plants. - - - - - - - - - - - - 161 

OSCAR LOEW: Ueber die Wirkung des Urans aufPlianzen. - - - 173 
K. ASO : On the Physiological Influence of Manganese Compounds on Plants. 177 
K. ASO : On the Action of Sodium Fluorid upon Plant Fife. _ _ . jgy 
K. ASO : On the Action of Sodium Silicofluorid upon Plants. _ - _ j^j 
S. SU2^KI : On the Acticni of Highly Diluted Potassium F>Jid on Agri- 
cultural Plants. lyo 
S. SUZUKI : On the Poisonous Action of Potassium Ferrucyanid on Plants. - 203 
K. ASO : On Oxidizing Enzyms in the Vegetable Bod v. - _ _ _ 207 
S. SAWAMURA: On the Curing of the Kaki Fruit. - - - .. 237 
K. ASO : On the Difteient Forms of Line in Plants. - . . _ . 739 
T. TAKAHASHI : On the Alcohol Production in Phxnogams. - - - 243 
S. SAWA : Can Alcohols of the Methane Series he Utilized as Nutrients by the 

Green Plants.' -------.___ 247 

C. KIIMOTO : On the Occurrence of INIannan. _---._ 253 
T. KATAYAIMA : On the Cicneral (jccurrence of Bacillus IMethylicus in the 

Soil. -----------__ 255 

S, SAWAMURA: On the Liciuefiction of Mannan by Microbes, - - - 259 

T. SUDA : Chemical Note on a Singular Phxnogamic Parasite. _ _ _ 263 

S. SAWAINIURA : On the Action of Formaldehyd on Pepsin. - - - 265 

O. SHISHIDO: Ueber die Einwirkung des Flara-Brennens. _ _ _ 267 

K. HEFELE : Die zukiinftige Bewirtschaftungsform des Japanischen Waldes ! 333 

K. IIEFFI.E ; Walil und Wasserwirischaft. - - - - - - 345 



[ ii ] 

H. SHIRASAWA : Ueber Entstehung un 1 Verth diking des Kamphers im 

Kampherbaume. - - - - - - - - - - "373 

S. SAWAMURA : Investigations on Flacherie. ------ 403 

O. LOEW und V. KOZAI : Zur Physiologic des Bacilla> pyocyaneus, II. - 449 
M. TOYONAGA : Ueber den Kalkgehalt der IMilchdriise. - - - - 455 

O. LOEW : Der Erntequotient. - - - - - - - - - 459 

O. LOEW : Ueber die physiologische Wiikung des Chlorrubidiums auf 

Phaneiogamen. - - - - - - - - - - -461 

M. NAGAOKA: On the Stimulating Action of Manganese upon Rice. - - 4^7 
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. - - - - - 481 

S. SUZUKI : Can Sulfo-Derivatives of Hydroxylamine Serve as a Source of 

Nitrogen for Plants.^ - - - - - - - - - -491 

K. ASO : On the Influence of a Certain Ratio between Lime and Magnesia 

on the Growth of the Mulberry-Tree. ------- 495 

G. DAIKUHARA : On the Influence of Different Rados between Lime and 

Magnesia upon the Development of Phaseolus. . . . _ - 501 

G. DAIKUHARA : On the Behavior of the Phosphoric Acid in the Soils 

towards Different Organic Acids. -------- 505 

]\I. NAKAMURA : Can Boric Acid in High Diution Exert a Sfunulant Action 

on Plants ? ----------- 509 

S.SUZUKI: On the Action of Vanadin Compounds on Plants. - . - 5^3 

S. SUZUKI : Can Potassium Ferrocyanid Exert any Stimulant Action in the 

Soil on Plant Growth .^ - - - - "5^7 

S.SUZUKI: AreSolubleIodidsAbsorbedbytheS.il?- - - - - 5^9 



In wie feme 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"-^ 

■^'-x ^ y^z '^z^ x^y 

also Functionen der Verschiebungscomponenten u, v, w darzustellen 
suchen und dabei untersuchen, wie ein krystallinischcr Korper, wie Holz 
als ein isotroper Korper betrachtet wcrden kann. 

Die Krafte welche die Theilchen eiiies dilatirenden Korpers in ihre 
Gleichgewichtlage zuriickzufuhren streben und als Driickkriifte wirken 
haben ihren Grund in den Kriiften, 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 aiis der naturlichen 
Gleichgewichtlage verschoben worden sind. 

Unter dem natiirlichcn Zustand eines Korpers verstehen wir den 
Zustand, dass ohne Einwirkung ilusserer Krafte alle im Jnneren des 
Korpers thiitigen Krafte sich gcgcnseitig aufhcbcn. 

Es seien die Coordinaten eines Massentheilcheii dm z y z, die des 
zweiten Massentheilchen dm aber .r + ^ / + ^ -+C' u'^<^^ cs wirke auf den 
Punkt {xy.z) die Kraft 

dvi dni'/if") 
langs der Linie p— \/^' ^-^^j^^^ 

die Resultate aller auf dm wirkenden Krafte ist 



= <-////; I / {f>) dm' 



wo das Integral so weit zu erstrccken ist, als _/"(/?) nicht verschwindet. Es 
ist sonach im naturlichen Zustandc 

* Franz Neumann: Tlieone cer Elabticitat pag. 'Sj. 



D. Kitao: 



o = dm \ diii'fip) — = dm \ dm'fif*)-^ = dm l dm'f{p)— 

Nun erleiden die Theilchen (.I'j-) und7/ + c, yArTj^ - + C relative Verriic- 
kung, deren Componenten durch 

^-' -y r 

bezeichnet werden mogen. Die Componenten der auf dm ausgeiibten 
Molecularkrafte sind 



dm \ A dm' — dm \ din'/{(> -h o' ) ^ , , 
J J f'-^P 

dm f B dm' = dm [ dm'/ (/> + />')-|4^ 

(f/w 1 r dm' =dm\ diJi'f{p-\-p')^—^ — -. 
J J ' ' P+P 



'o p' durch 



p+p'=V{^+^'r+(r, + -r/y+Q+:'y 

definirt ist, oder mit Vernachliissigung unendlich kleiner Grosse hoherer 
Ordnung 

mit dcmselben Grade der Anniiherung geschrieben werden kann 

/(."+^')(|±{;)=(A(,o)+^//'(,")(4±g\ 

p ' 

^-{/iP)^P'/(p)){^rn(^l-'^) 

= ^fip)^^^'^j;{/i^^-^f^^' 



Wo 









P dp 

gesetzt wordcn ist, so folgt. 



dm f A dm' = dm [dm' (^ f[p) + ^~^^' + ^ Hp) pp') 
dm f B dm' = dm f dm' (j^/ {<>) + ^ q' + -q F(p) pA 



In ivie fern kann man das Holz cf e. ^ 

In wie fern kann man das Holz. 

dm f r din' = dm [dm'(^f{p)^^^::+-: F{o) pA 

Es werde ferner auf die Oberfliiche des Korpers ein Druck auscreiibt, 
dessen Componenten fiir die Flachenheit 

S H Z 
heissen. Es seien ferner on ov ow unendlich kleine virtuellen Verriic- 
kungen d2c ov ow ihre Werthe an der Oberfliiche, deren Elemente dco 
sein moge, das Princtp der virtuellen Verriickung giebt. 



H do- + H (jv-}-7a div) dio 


+ dm dm ' A on 


+ 1 dm dm' B oiv 


+ dm I dill r oiv — o 


\ dm I dm' A on 



Mit dem Intecfral 



nehmen wir eine wichtige Umformung vor. In diesem Integral kommt 
die Wirkung des dm auf dm' als diejenige des dm' auf dm vor, und beide 
Wirkungen unterscheiden sich im Vorzeichen. Die Coordinaten 

^ + ? y^-q .C' + C 
verwandeln nach der Verschiebung ?/, z/, %u in 

,. , . ^v -- ^ 'dv „ , 3?.' 

ox ' dy - az 
Da ^ ;^ " unendlich klcin scin sollen, die Grosscn 

*3« , 3;/ , Jdu 

'a? +'-33;+' a; 



I). Kitao: 



d.v ' oy oz 
sine! iiiclits anderes, als die Componenten der Verschiebung, welclie die 
beiden Punkte x y .c, und x-\-z y + '^, -c + C nach der stattgehabten Ver- 
schiebung gegen einander erhalten haben d. h. 

Es ist so dann 

^ / az/ . 9zy\ ^ ^ /aw 9;/\\ 



Oder indem wir setzen 



Wo 

sind, so folgt 



/,,= 



/.,■-.: 



4= 



„. fip) 



dm 






dm'r/:: 
din'y^' 
dm'fZ 



f{f>) 

Zip) 

(> 



O 

I 

dm r^ 



>.--[ 



l,,^\dm'C 



fip) 



/,^ _ /._,| /,., _ Ym /-^ — h-2 



In wie fern kann man das Holz otc. 5 

Delint man das Integral iiber den ganzen Korper aus, so ist es offenbar 
dass wenn wir den Coordinatenzuwachs fiir dm mit abc, denjeni'gen fiir 
dm' mit a' b' c' bezeichncn 

\din\dni' A ^)// = i| \din' dm k {da — ua') = '^A\d]itdin'k. d{a — a') 

= d[dmdin' \{i)'=') 

da ja 

^' = a-a' r; = b-b' :' = c-c' 
sind, wir erhalien somit 

\dii' (H OH + H ov + Z ozv) 

-vddvAdm'Xd^' 

+ \[dm[dm''Bdy/ 

+ ddm [dm'VoZ' 
oder in dem \vir fiir A B F ihre Ausdriicke einsetzen 
\ (H o?^ + H dv-\-7i ozv) dzv 

+ -i f f dm dm' ^^ (,-o\-' + r^^rr; + : J^') 

f ^ [ f ^;;^ dm! ^^^(C'oT +;;oY + C'>\:') 

+ 1. f frt'//^ dm! r {<>) no' (,-J,-' +■-;'>/ + r<') = ^ 

Da nun aber 

o{p(!) — ^(j^ +rj<jr^ +^0^ =P<jp 



so folgt 



I dtu (H die -f- HoV + Zozc) 
+ ^[[dmdm'-^^pp' 



D. Kitao: 



— 1 1 dm dm' F{p) p'f/'- = 



■ +4 

Es sind die Verschiebungscomponenten des Punktes -r / ,c, ?/ v w sodass 
seine Coordinaten nach der Verschiebung' 

sein werden. Es gelangt der Punkt \n 

X + !/-{--' 

y + v^r/ 

- + t.' + c' 
Es ist so dann 









Da ferner 



. ^- / 9?/ 3// 3z; 3z' dzv 3zv \ 

'^'^\dx dy^'dl- 37 +3J'^J 

.,f 'dit 'dii 3z/ 3^' . 322/ Szi* \ 
.^^"f 3// 3?^ 3z-' 3z/ 37C' 'diu\ 

■^^'V3J' a?"^ aT' a^"^ 3?' 3J/ 

I , / 3// 3?^ 3^/ 'dv _3tc' 3«/\ 
■^^'^\3.r 3)7+3:;^' S^'^SJ' 3^; 

■^^H3J' 37+ a^' 3J+ 3^' a? j 



d 



In wie fern kann man das Holz etc. 

da ferner 

3// 3,1' , 3z^ 3?/ \ 

/ J3z^ 3^' ^ 37' 3z/ , \ 
+ l'3J'3J^=^ + ^^'3J'-<) 

^ iH 3^J +n 37; +^ ( ajj +-3F' 37' 
3te' 3z/ 37C/ 3w \ 

+'a?'3J'-+^37'aF'-<) 

+ ^'T at' 3,t-+''^37' 3J+^' 3F' 3F 

, * a« 3z/ 9 3// 3i' ^'dv 3// \ 
"^^'^3:^' 3j.+^^^'^+''^'"37' a?j 

+ ^"aF'3l+^-'-37'37+C-3j.3jj 
I ^ ^^/ir2 a^^ a// ^ 3?/ 3z£; ^ 3;/ 3z/ \ 

^^"H'^'a^+^^^a^+^^^aFaFJ 

+ "^3I' 37-^^37^+=?-37-37j 
+ ^'3J' 37+^^' 37' 37+^-3:- 3FJ 

+^n^ a?' 3J ^^'^s^'aF+^'^a?' 37 
+^^3.'3j + ^^37'ar-+-3J'-a^) 
+ ^^^'ar-+'^.37^+'^^^^'-3F) 

'^^ 3,v' 37 + ^- 37' 3? +-^-37' a? j 

indem man die Multiplication ausfuhrt. und nach dem .-* und r/ und etc 
ordnet. 



D. Kitao: 



+"^Vva^ +Vav;+-a.-' a- +-a;r' a^y 
"..//a?' V , /ar(y\ ary az' , ^at^ ar^A 

. f.. ( 'du du , 3u a?/ \ 

^5„/ a« a« a^^ a^c/N 

o^/ a^* a:' , dc' azc \ 

. ^,^/ a-cc are' , a// arc \ 

^3 / a^f^ aw „ a^^ aze; \ 
'^"'■^r'a/' aJ^ a^' a^r ; 

*.o„ / a// a« a^^ a^,' , a« azc 

+ ?-C''J 2^^,-^— +2-^^—,-;^ 1-2 

'V ay a^ ds ax 



a^'aJ' '^^aJ''a^ 

a?/ az/ a?*! az£/\ 
"^^a^' aJ"^^~a;^' a7/ 



,^ / az^ az/ az; a« a« dv . a« a^ 
+'^"^'^raI'aJ+^a7' al+^^' a^ "^^aj' aj/ 



a« azf a?^ 3zv\ 
'^^a7'a7'^"a^''a^/ 



, ^,^ / du dw , a// az^ , d2a du , ^ du 6iv 



a// azc; a?/ azc/\ 
+^ai?' a^'^^aJ' aJ; 



Setzej^man ferner 

2^] 



= U;/ /'^ (/>) .-' 2a.,^ [dvi' F{f>) r/ 2a,^ \dm' F {j)) C 
2a, = \dm' F ip) qhj 2a, = {dm' F (p) r/: 2a, = [dm' F (p) f? 
= [dm' F{p) ^f 2a,-= [dvi' F ip) C'-y 2d'= [dm F{p) f^C 
^;^=[dm' F{p) Cy 2a,,= \dm' F [p) ^T' 2ay,= {dm' F{p) ^'vj 



2/3'7 = 






In wie fern kann man das Holz etc. 



2a^,= 



'=Um' F{p)^"^: 2a,,= \dm' F {p) ^-^X 2a,,= [din' F {p) ^-rj^' 

+ K(a7+ar)+^a?a7)) 
+4M(aF+i57AaF+37J + 9:7(a7+a?)) 

// 3« , 3r' V3^c , dv\ , dv /dw , 3?/ ^^ 

+M( aF+aJ A 97 +3j)+ wl a? + ajjj 

// 3^' , dzu\/'dzu , 3// \ , dzv/dzi , 3t/ \\ 

+4^(37+157X37+37)+ 37(97+ arjj 



Man setze ferner 



so erhiilt man 



dP = d\ dm \ dm' f (p) p' 

d Q = d{dm {dm'/^{^'--{-r/' + C'l 
o R = d\ dm I d7u ' f (/>) p- p''- 



, 33v J 3dv , 33:7 
d.r d/ o^ 

, 3 ^w ,33 oza J 3 <?t£/\ 



Setzt man ferntr 



3u J 3w , 3« 



, 3;/ , . 3;/ « 3/^ 
dx dy 3:7 



lO 



D. Kitao: 



3z/ , . 3« . , 'du 



^/3 = /3i9j + /-3.;3^+/.S3^ 






■ ax ay os 

, 'dw J 3r£' 3 ^iv 

. 3z£/ . 3zt' , , 'dw 



So erhalt man 



3(5// 3o7/ , 3o7/ 



> ^ f 7 ^ / 3d// , dou dot 
dov dov , dov 



3(5zc/ , ddzu , 3ozf \ 



Setzt man endlich 
o 



3?/ , dv , 3w , /3y ,3w\ /3zc' 3//\ 



/ 3// , 3v \ 

+H37+a?J 



„ 3u , dv , d'-dJ , {c)v , 3w\ , /3zi' 3«\ 

n,,=/z.,^+/^,^ + /r:o-3j + ^,(^3j^-g^j + ^u(^3^ + -3j; 

^ /3// , 3v\ 
3// , at; , 9w , /3r' , 3t£/\ , /3w , a//\ 

n,.=/^n 3- +^,0 3^ +^33 97+^8 (^ a7 + 37 j+ ^« V 37 + arj 

/ 3// 3y \ 

+ H37'^3F; 



„ ^ 3z/ , 3?; , 3zc' , / 3v , 3w\ , I'dxv 3// \ 



/ 3// 3t; \ 

+H37+aTJ 



I 



Ill wie fern kauii man das Holz etc. 1 1 



/du dv \ 



9/ dx ) 
„ „ 02/ 3z' , 9z£' , /97.' , 9j:t^\ , /3w 32/\ 

, (du dv\ 



So erhalt man 






I o ^^'''4-0 ^^''' +0 ?^ 



9ow „ 'dozu „ 3Jw 



) 



Wir erhalten somit als Gleichgevvichtsbedingung 

f (H on + H 02/ + Z otc/) + 4 o/' 4- -}S + i oR = O 

Wir untersuchen die letzten drei Integrale besonderes, indem wir setzen 

£/j;i = iidxd:::dz = jidr 

und sclireiben in dem Integrale 

'doll 



HH'"^ 



sodass 



, dou dulxxoii dalxx -> 
alu^^- = — -^ ^ — o u 

ox ox ox 



[,„,,l^..=[a.^^_P.JM, 



ou 

dx 



in dem wir das erste Integral nach dem Gaussischen Satze transformi- 
ren, so erhalten wir 

r , dou r - ^ r 7 dfjiXxx ^ 

\ f-'y^ "o~7""= — 1 {^^^w ^>u cos nx— u/r-^-7-0// 
so mit alien andercn Integralen 

^ ■^= ~" I diofii y.j^ cos jix + ?^xi cos 7iy-\-i^iz cos ;/~~ j<5« 

+ ( /wi cos 7/.r + /,.2 cos 7iy-T L^ cos ;/~r j^z' 



12 



D. Kitao: 



-rl ^1 COS 7LV-{-?^22 COS //J' + ^.-j COS J/S jOW 

I 3.1- 3/ 3,0" j 
Wobci das Oberflachenintegral iiber die ganze Oberfliiche des Korpers 
auszudehnen ist. 

Die Grossen ?. sind im Jnneren des Korpers constant. Wir konnen 
sie aber . nicht unmittclbar an der Oberfliiche constant vorstellen, da 
in dem Integrale ?. um so mehr Glieder felilen, je niiher der Funkt .lyrj an 
der Oberfliiche liegt. Die Grosse ?. muss also so beschaffen sein, dass 
sie, da der Wirkungskreis der Molecularkriifte sehr klein ist, zuerst 
schr rasch variirt mit wachsender Entfernung von der Oberfliiche rasch 
gcgen eine constante Grenze convergirt. Das Raumintegral braucht 
daher nur liber die der Oberfliiche unendlich nahe liegende Theile 
aiisgedchnt zu werden, wenn der Korper homogen ist. Das Raumintegral 
denken wir uns in zwei Teile geteilt, eins iiber den Theil des Korpers 
ausgedehnt, innerlialb dcssen / Constant ist, das andere iiber den iibrig 
bleibenden Tcil in dem letzten Integral setzen wir 

dt=dco X dn 
dz = dco X dn 
wo dn ein Element der Noimale n bedeutet, fil ist demnach cine Func- 
tion von n, o). Bei der Klcinheit der Grenzen von ;/ konnen wir wohl 
ohne Bcdenkcn, mit Neumann annclimen- dass nX cine Function von ii 
allein sei, cine Annahmc, die niclits andcrcs aussagt, also dass /A in 
derselbcn Ticfe untcr der Oberfliiche iiberall denselben Werth habe 
Wir kcinnen, in sofern dieses erlaubt ist, setzen 

'did 'dul 
—^ — = — 75 — cos UIX) 
ox on 

so dass wir schreiben konnen 

f , f . 3/i/in , 

-on cos nx 



I dz -—-on = \ do) \ dn ' ^ oi 
J ^-^' Jo J 3« 



• Thcorie der Elnsticitiit 9G. 



In wie feru kaiiii man das Holz etc. i ^ 

wo 71 = der iiusscren Oberfliiche entspricht. Particulilrc Integration ergiebt 

f , > , r \ { J { J ■i do cos nx 

= — yiio oufxK^x cos \ii-'^)— \ do) \an}xh^^ ^r-^ — 

+ I do) oiifAiy cos {lix) 
Mithin erhalten wir 

o P= — \(Hoij. i (,^-n COS ;/x + ^.,2 COS 7//4-^T, COS (;ag-) Vm 

+ ( L\ cos IIX + /w2 cos «/ + /w-, COS (//^) jot/ 

+ ( ^'31 cos ;/.f + ^2 COS ;/;' + ^.33'cos {ji.-:;) ynu 
, r 7 r 7 /■) dou COS 7/x , , doll cosily , . dou cos{n::;)\ 

+) '^"' j'^"K'-— 1& — +^" — ;7;r^+^-" ,// ) 

, <r/or' cos «j' . ^ot^ cos (;/^)\ 
"" d}i '' dn ) 

J dozu cos ny , rtf^zc/ cos («^)\ 

+ •^"- 7bi + ^'^^ 'd^i ) 



- dov COS mi 

+'■' — li. — 

. doZV cos 71 X 



dn 

•)// 7)11 . 7)n \ 

l + etc 



.J,..(,,|^,;,,|^,;,„|^). 



Die Dichtigkeit /i kann inncrhalb der Molecularentfernung sicli iinderen. 

Wir woUen den Korper so beschaffen vorstellen, dass eine Volumen- 

cinheit iiberall dieselbc Menge Korper enthaltc, d. h. so ist die inittlere 

Dichtigkeit iiberall constant und der Korper ist homogen gebaut da 

feriier X in dem ersten und dritten Integral wesentlich constant = i7, so 

verschwinden die Intcgralc. Wenn wir ferner die Annahme machen, dass 

man von clem Zustiinden der Theilchen die unendlich diinne Schichte der 

Korperoberflachc absehen konne, dass die Theilchen dort iiberall dieselbe 

Verriickung erleiden, als wcnn die besagte Schiciite starr wilre. So ist 

die Grosse wic on cos {n.\) von n unabhiingig, so verschwindet das zweite 

Integral, Mithin ist iiberall 

oP = o 

in so fern als der Korper h.omogen ist, und man von den Zustiinden der 

Theilchen in der Oberfliiche abstrahiren kann. Selbst aber. wenn der Korper 

heterogen ist, so verschwindet ol\ da w Giossen /„ etc in dem inneren 

Punkt verschwinden, wcnn cs nur von den Vorgiingen in der Oberfliichen- 



J. D. Kitao: 

schichts abgesehen werden kann, Fiir den Ruhezustand des Korpers in 
sei'nem natiirliclien Gleichgewichte d. h. ohne Aiissere Kriifte muss oP 
unter alien Umstanden verschwinden, dann wenn 

H = H = Z=:0 
so miissen auch 

U ^= O =^ V =: Zi' —- o 

da nun ou uv, uzv ganz willkiirliche Functionen sind, so wohl im Inneren 
als an der Oberflache des Korpers, so miissen 

etc 
/•u cos (;/,f) + /io cos {fiy) + lyi cos (;/.c) = o 
etc 
d.'h. aus den letzten drei Gleichungen folgt, allgemein 

/j J =: /j2 = /i3 = /oi ^ /22 = ^23 = /31 = ''•Si ^= ''•;« ^^ ^ 

wodurch auch die drei ersteren Gleichungen befriedigt werden. 

Um oO zu untersuchen nehmen wir dieselbe Transformation vor, in 
dcm wir setzen 

Mithin 



■uu 



''(?= — 1 /''c/"^* ( ?/i cos ?/.i' + 7/0 cos iiy + ?/3 cos (;/.:) \oit 
+ ( z^i cos nx^-v^ cos ;;/+ :'j cos (t-.::) )ot' 
+ ( zc'i cos ;/.r + zv. cos ;/y4-^t'3 cos {u.::)jdzu 

J \ d-v dj/ d^ / 

/ a/izc;, a/7t£/2 9/^+A > ., 

das Raumintegral braucht nur iibcr die der (Oberflache uncndlich nahe 
Schichte ausgcdehnt zu werden, da //j //■_. u-^ etc. fiir jeden innern Punkt 
verschwinden. 



In wie fern kaiiii man das Holz etc. i r 

Wir denken uns an der Oberfliiche eine unendliche diinne Schichte 
von der Dicke n innerhalb deren I von o sich unterscheidet, und aus 
dieser Schichte ein Prisma ausgeschnitten, dessen Grundflache dco unend- 
lich klein, aber unendh'ch gross gegen die Hohe ist, dieses Prisma soil 
in seinem Theilchen iiberall dieselbe Verriickung erleiden d. h. wie ein 
starrer Korper sich verhalten. Wir bilden dieses Integral iiber dieses 
Prisma und vernachlassigen dabei die vier Seitenfliichen gegen die beiden 
Grundfliichen da nun lu. nur von n abhiingen soil und in jedem Theil des 
Prismas 7i v zu dieselben Werthe haben, so hiingt jiit etc nur von ii 
ab. Es ist daher 

—k — = L cos inx) 

ax ail ^ ' 

^ ' -— ;C ' cos iny) 
oy on 

'dutio 'dun-, , 
—h: — = /, ' -cos (;/.c) 

Und wenn wir wieder setzen 
erhalten wir 



dT = dio dn 



-\ —. r 



f,_ / 'dan 

f , [ '] J \( (^I'-J'x dull. , duu-^ \^ 

= I d(.o \ {dii)\ — - — cos iix H — ^3-^cos ny-\ — ^-^ cos nz joii 

— \ doj ui 7t\ cos ;/.r + //o cos ny-\-ni cos n::\du 

— I dco (1 u\ cos 7/.1-4-7/0 cos //j' + .Vo cos na\uu 

das erste Glied bezieht sich auf die Oberfliiche das zweite aber auf die 
innere Grenze, weil weder cos etc noch nu or ow von ;/ abliLingen soil. 
Bildet man weiter auf dieselbe VVeise die Ausdriicke 






so erhiilt man 



1 6 !)• Kitao: 

^Q=\fi dcoiiii cos fi.v + u-, cos //7 + ?^3 cos ;/.c- j'^''/ 

4- l/z rt^'ctif z'l cos ;/.r + 7-'o cos ny-^i'-;, cos nz\oy 

+ la ^oj( tc'i cos ;/.!' + tt'2 cos ny-{-wl cos /ac )o"t' 

Wo die Grossen sich auf die Punkte beziehen, die sich auf die innere 
Grenze der Oberflachenschichte beziehen, da aber / an dieser Grenze 
verschwinden, und mithin auch «, so folgt 

da dieses in jedem beliebig liegenden Elementarprisma stattfindet, so 
muss dies auch dann stattfinden, wenn die Fliichenintegrale iiber die 
ganze Ausdehnung des Korpers ausgedehnt wird. Mithin 

iiberall, in so fern man von den Vorgllngen in der iiusseren Oberflachen- 
schichte absehen kann. 

Das dritte Integrale oR wandehi wir auf dieselbe Weise um, in dem 
wir setzen 

ox ax ax 



so folcrt 



oR— — \n dU (i>„ cos 12X + iiy.^ cos ;;_;' + i-^3 cos //c) yyu 
+ ( (X^oi cos 72X + IJ..., cos ;// -f //.3 cos }i::)jdv 
+ ( (i^i cos 7ix 4- i^3j cos nj' + —33 cos nc) j'>^«' 

]' \ dx dj d:y ) 

V ax dy d.z J 

Wo m "11 -Vi. etc in All<7cmeincn die ractorcn von -75 75— etc andere 

'^ dx dy 

Wcrthe habcn wcrdcn als in /.^n /?ij etc da die Grossen ai a-, etc im Jnneren 



In wie fern kaiiii man das Hoi/ etc. J7 

constant sind, aber an der Oberflachc ini allgeaieinen Variabel sein 
miissen. Wir zerlegen das Rauinintei^ralc in zwei, das eine ausgedehnt 
iiber die Oberflachenhiille innerhalb deren ^j a. etc variabel sind. Und 
das andere iiber den iibrig bleibenden Raumtlieil. In dem ersten Integral 
setzen wir wieder 

d~ = dio dii 
und 

-,^-— = —- cos nx^ — : = ^- — i--cos ny 

ox dn dy dm ' 

etc 
da nach der eingefiihrten Annalime jeder Punkt des aus der Grenzhiille 
ausgeschnfttenen Elementarprismas a Is starr betrachtet werden kann und 
die Grossen a^ a-, etc von n allein abhiingig sein sollen. Das in der 
Rede stehende Integral, wird, on cos vx etc von n etc nicht abhiingig 
sind, 

\dio lA (.(?,! cos iix -f /?,. cos 7/J+ .(?,3 cos na) \du 
+ i (.(?ii cos iix -i- il» c OS 71)' + /A3 cos ;;.c) \dv 
+ ( (-'-^31 cos ;/.r + /j'32 cos 7iy -f- .(^33 cos w-c) \8w 
— \dco //( (il^ii cos nx-\- iiy, cos ;/j'+ ^^13 cos //.:.) )Jz' 

+ 1 (i/,l cos 7lX-\- il^., COS 7lJ/-\- SJ.. cos ?2S) joz' 

4- ( (/i/32 COS 7.'X 4- SJ^o COS ;/;' + 1^33 cos //^) jozc/ 

durcli die Einsetzung dieses in oR licbcr. sich die Integrale auf, in denen 
ii^n X^j2 etc treten so dass 

''A'= — \dci> fi (.(?,i cos 7/.1- + .(?,._, cos iiy + /-^i3 cos ;/^) \bu 
+ ( (i?2i cos 7ix + //.._. cos /// + //.. cos ;/.r) V)i' 
+ [ (/?3i cos A'.r + /? 2 cos_//j' 4- /.''ai cos ;/:;) \bw 



J 8 D, Kitao; 









Wir erhalten als Gleichgewichtsbedlngung 



!- 



da an der Grenehiille 

{{JU = OU dv — 01' O'ci^' = ow) 

Wir haben somit als die Bedingung ftir die Gleichgewichtslage 

/J. i?n cos 7/.V + /X <Jy2 cos /// + « -'?i3 cos n.c) — H 
// i^ai cos 7/.i- + />{ /42 cos ny+ix il,^ cos ;/.s) = H 
H i?3i cos i!X+(J. /4> cos ;/>- + />« /^s cos nz) = 7i 

a^ yj/ a- 

3r 3/ 3- 

'dx d_y 3^ 

Vergleicht man mit der Gleichgewichtsbedingung 

dXx , 3X>/ , 3A'^ 

-I ?=r'^^-^ ^ = ^ 



3.t' 3/ 

dVx dVy , 3F^ 



3,r 3j/ 3.c^ 

3Z.r 3Zj/ 3Zcr 

■H ?=e^- i ?s = ^ 



3.t' 3;/ ' 3^ 

so erhiilt man unmittelbar 

Vx = n /4, 1 > = /< //.2 5 '- = /^ -^-^si 
Z.v = ,« /4. >^7 = /^ -'-'32'. Z^ = /^ ^^^z 
/ du , 37' , 3w ^ /37/ , 3t.'\ ^ /37c' 3?/\ 

,/3w , a-' \ 



In wie fern kanu man das Holz etc. 



19 



/du dv \ 

,, / dti , dv , die , ( dv , die \ , / dtu , du \ 

( du dv\ 

T^ / du , 3^^ , 9:i:' , / dv , div \ , / dzu , 3z; \ 



, / 3^^ , dv \ 



Tr / 9^^= , a^^ , a^c , f dv , dio \ (div , a?/ \ 
^^='"r''-aJ+^^'-a7+^^'«^F+^4ar+-5rj^Ma:r+arj 



, , a?/ 3z/ \ 



■r r / du , a?^ , dzv , ( dv , dio \ , / a^c^ , a« \ 
^'=^=K^-^+^--a7+^^-aF+HaF+a;rj+HaF+-aFJ 

/ az' a// \ 
+MaF+-^j 



Z.= a/4 



-7 ( du . dv ^ div , /ar- , ate' \ , [d-iu , 3/^ \ 

, / du du\\ 

die 15 Coefficienten a sind die sogenannten elastischen Coefficienten, und 
sind Constanten ausgenommen die Punkte der unendlich diiniicn Ober- 
flachenschichte. 

Diese Functionen lassen sich anderes schreiben, in dem wir iibcrall 
einfiihren 

du _ _ dv _ ^ div _ ^ dv dzi> _ arc . du _ _ 



dx ~^ aj ~^^ ds ""-■-■ a- • dy ~^' dx ^ dz 



du , dv 



dy dx 
^x =/'«(^i X, + ihi J V + '^11 ^z + ^13 - tf + ^9 -.- + ^?4 ;'x) 



20 



1). Kitao: 



^'v = /K^h- -IV + (^-2 Tv + ^10 -.- + ^5 ;'-- + ^\i -^'c + ^7 J'^) 

Z- = /i(^u ,r,. + ^jo J,, + ^73 ■■"-' + ^S J'. + ^C -^'c + ^15 7^) 
Zj, = 3 '- = ,«(^n + «!5j'</ + ^S ■^- -^- ^10 -J- + '^15 -^'-- + ^IJ Jr) 

V^ — A'y = n{ai -r, + a- j',, + <?i5 -c + ^u - i/ + ^u -r- + -^ij /x) 
Man bilde 

+ Cj'j,- + 2 C,;3>',^. + 2 C.^yyX, + 2 Ca^r^V + 2 (^^c J-/J'. 
+ 6^33.^/ + 2 C4.C.-- <, + 2 (TssS'.A-, -f 2 Q.C-,;'^ 
+ 6^4.7/ + 2 r45-2/.r, 4- 2 C^^,z„y^ 

+ Qc jjv 

unci differentirt man nach den 6 Argumenten 

■'^'x y,j -. -^r }'x '^'z 

Vy \ 



3j 



a 
a/ 



a 
a/- 



— = ^ CjX . 4- C-js;:-/ + Gs- ^ 4- Gi- ;/ 4- Gs-l'^ + GrJ'o- 

— = ( ^u-^'x 4- C-^i}',, 4- ^34"; 4- ^44- >/ 4- Cis^j; 4- c JO ,r.r 



aj 



Wenn wir hicrin setzcn 



— - = ^ <r,,,,iV 4- C,r,y,j 4- 6^,5-,- + C\.,-y 4- Q-r^ + ^'srj: 
— = ^ C^(,x, 4- C,;', + Go--- 4- Ci^.-y + (T;,;.;-, 4- C r.,;/ 



^7j = 6"! , ^4 = 6^„- a-; = ^'or, 
^. = Cw ^5 = Cj , «s = C-ii 

^7lO= 6-3:= C/44 ^1.',== 6i4= C 15 

//.l = C^J3= W,ri 'i"!! — 2 6 v'5 — =6 40 



SO folgt 



X = /^ 



-dP 

dx^ 



V ap 

dy. 



^==''^-a?; 



i 



In wie fern kann man dsis Holz etc. 21 






d. h. die Coinponenten der Druckkriifte haben also cin Potential. Die 
15 Constanten lassen sich im Allgemeinen theoretisch auf eine geringere 
Anzahl nicht zuriickfuhren. Sie lassen sich, wohl, wenn der Korper eine 
gewisse Symmetrie in seiner Struktur besitzt d. h. ein Krystall ist, der 
nicht dem trikh'nischen System angehort. 

Wir nehmen zuniichst an, die Struktur des Korpers sei S3nTimetrisch in 
Bezug auf eine Ebene, nehmen wir diese zu (x .7) Kbene so verschwinden 
von dem Constanten a diejenigen in denen j/ in ungerader Potenz, vor- 
kommen, wie 



2 CZ4 = 1 (/7n' F {p) ^^y 



So ist in diesem 

ai = o a 3 = a- = o a<,^o ci\z^=o cix^ — o 
Mithin 

C\(, — C-2i=(^-26=(^Zi = ^ C^^ — o C^f, — o Ci^ = o Ci-^ = o 
die Funktion P wird in diesen Fall 

^ = h{CyV^r + 2Cy.x\y,j + 2C,^,x^z-, ■\-2C\-^x^x, 
+ C.^^y,; + 2 C.^^yy-, + 2 C.yX^j::, 
+ G3-/ + 2C,5:r,A-, 

+ Cc^y/ 
In der That bleibt diese Funktion unverandert. Wcnn man hierin statt 
yv ~-y — v setzt, denn es werden 

■^'■'~ \ acr • 3;/ J ^'~ \-dx ^ dy J 
die Anzahl der Constanten reducirt sich auf 9. Ist der Korper noch 
um eine Ebene etvva y.:; Ebene symmstrisch gcbaut, so verschwinden alle 
von den a in dcnen x in ungerader Potenz auftritt. Es werden noch 

a^^aT = iTr, =^ ^14 = o 



22 , D. Kitno: 

die Funktion P wircl in diesem Fall 



Wobei 
Mi thin 



C 2j — 644 C 15 — C35 6 J2 — Cpi 



+ Cvl2x^y,j+y;-) 
-{■ C. -,{2 y,j-, +::,/) 

Die Anzahl der Constanten so ist 6. Die .^'>' Ebene ist zugleicli audi eine 
Symmstriebene, dann P bleibt unveriindert wenn man .z mit — z vertausclit. 
Ein soldier Korper geliort dem rhombischen System. Ist der Korper so 
beschafifen dass die Struktur um eine Axe symmstriscli ist dass die Drudc- 
componenten sicli nicht iinderen wenn man z. B. x mit y vertausclit so 
wird die Anzahl der Constanten nocli geringer. 

Sei der Korper symmctrisch iim .r-Achse, so ist dann 

ai = a., ^16 = ^11 
{din'F{o) c^= [dm'F{p)Tj* [dm'F{fj)Z'rj'= [dm' F{pX''^' 

Cji=L-. 613=^ 603:= 644= 6-,5 

Mithin ist ir.-Jicscm Fail 

P = i ((:„(.tv +JV) + C^-^ + Cyl2x^y,j +7^-) 

+ CJ 2x^, + 2;/„.c-, + .r.) ) 

Was sich in der That nicht ilndcrt, wenn .r mit j/ vertauscht. 

Solche Korper sind Krystalle des quadratischcn Systems. 

Ist der Korper so gcbaut, dass er zugleicli .z. P. um die j'-Axc s)-mme- 
trisch gebaut ist, dass man audi .:: mit x vertauschcn kann so muss 

ax = a;, av,-=a^n 



\dmF^{,:)^'=^{din'F{(>)C 



In wie fprii kanii man das Holz etc. 23 

^11^^33 L- 13=- 6)3 

Die Funktione P wird 

Solche Korper heissen regulare Krystalle. Die Druckcomponenten des, 
regularen Krystalls sind 

X^-y.^ Cu-Vx + C^yx + <^i2-^) 
Yy - fx{ Cn yy + C^^x^ + ^.)) 
Zz=lJ.{C^^z,-\- Cyiyy-^::^) 
Xx= fiCy^xy 
Y:: = !iCy,y- 

Wir betrachten jetzt Korper, welche dem hexagonalen System ange- 
horen, die Structur des Korpers soil so beschaffen sein, dass sie sym- 
metrisch ist in Bezug auf drei auf einander seiikrechten Ebenen, dass eine 
Drehung um 60° oder einen beliebigen Winkel in einen von der 
urspriinglichen nicht unterscheidbaren Stellung fiihrt. 

Es ist im diesen Fall 

die Hauptachse sei .c-Achse, und in Holz wiirde .c-Axe die Richtung des 
Longitudinalen sein. Wir fiihren ein zweites Coordinatensystem x'j/' ein 
welche gegen das alte x y um den Winkel y geneigt ist 

x-=^x'a—y'^ a = cosy/ 
y = x'^^y'a ^ — sm^ 
a- + /?-=! 
und daher 

u — ii'a — v'^ 

Es ist 



24 



D. Kitao: 



dJt (in n , <:^v ^., / d'/' , di> \ T 

dx dx' ity' \ dy dx' J ' 

dv du' 3, ^ ./<-/ , . / du' d7>' \ . 

<af7/ , dw ( dv' div'\ ( div , div'\^ 

1 du dw\ fdm' , dii' \ Idlu' , dv' \ 

[-dr+li]r)='H'dr+i5r)"-[-7jr+ii^) 

oder wir die leichtverstandlichen Bezeichnungen cinfiihren 
x^ = x,,'or+y^,'f3'-;/Ju,3 

J'^ = ( -t X -7;/' )«/3 4-j'^' («- - /3-) 

- ?, = - ;,« + 't'2 'I'z - {xS) a — Zyii 

Wir bilden die Ausdriicke 

dP ^, dP „ VP 

Xx^^a—, — yy=a—, — Zz—ti—- — etc. 

dXj,, dyy d.z\ 

und setzen die Werthe der Druckcomponentcn in neuen Coordinaten 
system 

Xx' Vy Zz' Zy' F^' Xy' 
.. , dP ,. , dP ^ , dP 



^ , dP y , dP ,. , dP 

' dy, dz'^ -^ dy,j 



Es ist nun 



~\ dx dx'^ dy dx' ^ da, dx' ^ uy, dx'^, dyy dx'x) 

dx^ 2 dy. 3., dy, dy, dz^ 

dx'z ay X dy X dx ^ dx -^ 

dx',, dx' ^ ' 
.V'. = .\;a^+F,,? + 2y.«;9 
und auf dicselbc Wcisc 

F', = ,\;y?^+F//-'-2PV/,9 



In wie fern kann man das Holz etc. 

Y, = Y, ={Y,-X;)a-i^X,{o?-^) (I) 
Y',= Z'y=-X,J+YM 

Es ist nun 

Ya = Zy = ^uCy,s, = /^X;,(.c'/. + x'J) 
Z^ — X,= II Ciyt\ — l>.Cyix\a - s'yjd) 

Die Einfiihrung dieser Ausdriicke in (I) 

a' 

^'" ={C,,a- 4- ^3,5-^) +j'',( ^,3/5^ + C3«'^) + «,5r',(C, - 6^3) + C^^.) 

«,3-t-',( C23 - ^,3) + .-',( c'osa- + c:,3,^') 

-vW(C3,5^ + «^6^,3+ C„(a^-,9^) -.y,^;?(G3- ^13)) 






r 
Wenn der Korper so gcbaut ist, dass die Constantcn von y unabhiingig 

wird wenn man die Coordiiiatcnaxen um y^ dreht. so wird bei der Drehung 

um y wieder sein 



26 



D. Kit.io: 



fji 



Z'2 






— 6 23^ s + 6 28j' y-h C i^X j 



So miissen sein 



II 



(:^i//-.a2Cu-<^n(«'--/5')2ri,(«'--;3')-^ [iii 

dicsc II Gleicluingeii erfiillt man (lurch die Annahme 

Cj3=C:>3 L-ll= L 12=^ 3^ 13 

und da 

a-+i5-=i 
ist. Mithin erhaltcn wir fiir cinen Korper, der um die .c-Achse symmetrlsch 
gebaut ist, wie bei den Holzcrn, 

P=l Cylx- +j'-) + 6'i3(.tV + - / + ^^x"z + 2; V-.) + C.sPz 

■\-Cy.{2x,yy-]ry2) 
dicsc Glcichung gilt audi fiir jedcn Krystall, der dem hexagonalen 
System gehort, dcnn die Werthe "^ sind die einzigcn durch die Glcichungen 
gleichzeitig erfiillt werden denn die letztcn Glcichungen von I und 
II lassen sich audi so schreiben 

[i^Cy, - (JrC'n + 3(«' -/^') C\t=o 
a^C^ - ^-Cn + 3(«" - fl') Cvi = o 



Ill wio fern kanii man das Holz etc. 

Addision giebt dcnn 

Aus den beiden Gleichungeii in III folgt 

611= 602 = 3^ 12 



(7i={ dnJ F(,o) ,-V. = [dm'Fipyr/ 



Wir fassen jetzt einen Korper ins Auge, welcher so gebaut ist, dass 
die Druckcomponenten sich nicht iindern, wie man audi die Coordinaten- 
axen legen mag, einen Korper von dem man sagt er sei isotrop. Die 
Funktion P behiilt immer dieselbe Gestalt, wie man audi nie Coordinaten- 
axen drehen mag, als wenn der Korper in jeder Richtung sich verliiilt, 
Avie ein reguliirer Korper. Wir setzen daher 

P = Cni^V +yi + -z) + C,lvi +_;'/ + -,-) 



und setzen zuc:leich 



C,^^-K{i+k'-\-k 



L ,0 = — • 



2C,.^-2k'K. 

Wo Kkk' gewisse Constanten sind, wenngleich nach dieses Definition 
k' = l sein sollte so lassen wir sie einfach bcstehen aus Griinden, die wir 
spater noch unten auseinander setzen wollen. Es ist 

Cn+C,.= -/c'K-K 
da K=—2Cy. 

C,,-VC,,= -k'K-\-2C,, 

Cn-C,,= -k'K 

fur einen isotropen Korper ist P = 0. Wcnn also k' verschwindet, so ist 
der Korper isotrop da in jcdeni um ^-Aclise symmtisch gebauten Korper 



-k'K=2{drx^'r;-F(p) 



28 D* Kitao: 

muss durch C5^ zum verschwinden gebracht werden, damit der Korper als 
isotrop betrachtet werden kann. Die ^'/] sind intermoleculare Grosse, und 
der Ouerschnitt des Holzstabchen muss darum so klein werden, dass 



2Yd^i'YF{n) 



verscliwindet. Legt man die .c-xA.chse so in die Liinge eines Korpers 
etwa des Holzes, und .i' und r-Achse so verringert, dass 

\di>i:F{f>}{y-yr)r 

verschwindend klein betrachtet werden kann. Ein Streifen Holzfaser ein 
Holzstabchen von verschwindendem Ouerschnitt kann daher als ein 
isotroper Korper betrachtet werden, die Funktion P wird 



die Grossen 



d. h. 






du 3v 'dw 



d,r dj/ 9.C 

^\^^ -dz) 

iindcren sich nicht, wenn das Coordinatensystem gerilndert wird, wohl 
aber die Grosse 

um dies zu beweisen fiihren wir die Hauptdilatation ).i L ?.- ein und bezeich- 
ncn die Cosinusse dieser Richtungen, welchc sie mit den Coordinaten 
schlicsscn mit 

'/i;?i/'i '^-•^Lv'-- '^mI^i'-.'. 
Hs sind uns bekannt 



dx 



■ = x^ = a^-Xy- + a.pj 4- «.;-7>.,'- 



In vfie fern kann ninn das Holz etc. 



29 



3zu 



■-=yy=fi{K+fiiL-\ fi^k 



- .:\ = aJi 4- a:/.., + r/.^-li 
oz 

\\^ 4- -^^J = -2" "" '''"'^^^ ^ ^-''— "^ ^'''''^'' 
-/ 3// , 37' \ , jv -I ■ - ■ . o - 

Kl7 "*" ar) + 2 = "''^'' ' + "^'''='-= + "'-^''-^ 

Wenn wir jetzt die drei ersten Gleichungen addiren, so konimt 

3// , c)v , cizv , , ... 

^— + -=r- + ^ := X, +y„ -f .^^•: = /i + /- + /s 
ox oj> orj 

was die Volumendilatation bedeutct und von den Cooruinationsystem 

ganz und gar unabhiingig ist 

Man findet ferner 

+ 2/ A(«i V + /5ni- 4- /'iW) 

+/!/(«/+ A' +r/) 

4- 2/i;.3(«, V + /^iW + /'i''''.i') 

+2^^(«,v+,9/,?3^4-r/?-.') 
+ 2;i,^(/9/jv+/-.V+«//V) 

4- 4^'i^-2(/5iri, Vr^ + «i/'i«,j-, + «,;5iao,9o) 

die letzten drei Glieder lasstn sich andercs sclirciben. Es ist 

«,«. f ;?,/9.4-;'i;'o = c' 
das Quadrat hiervon ist 

— {'h'(-'i + ^1 W + rrr-i) 



OQ D. Kitao: 

ganz auf dieselbc weisc 

Er folgt hieraus 

- 2/i^(«,^«/ + /5/^ - + riTi) 
-2;,;3KV+AW+riW 

die Addition dieses zu -t'/+j'/ + -/, so heben sich die mit ?^il2, ?.i?^z und 
hK multiplicirten Glieder auf. Mithin 

-^'^' +j'/ + .cr + 1 (-/ +j'^' + -i-;-') 

d. li. unabhangig von der Lage der Coordinatenaxen, in dem fiir P 
angestellten Ausdruck ist aber nach das died 

vorhanden, das im allgemeinen von der Richtung der Coordinatenaxen 
abhiingig ist. Soil aber P in jedem rechtwinkeligen System unverLindert 
bleiben, so muss 

k' 
vcrschwindcnd sein, d. li. irgend wie 

unendlich klein sein, damit der Kor[)cr ein Isotropcr sein konnte, welchem 
System cr auch angehoren mochte, 

Wir erhalten fiir eincn isotropen Korpcr 

P=- K{x,' 4-j'/ + ^V + Kj'--' + V +J'x') + /^-(-^'x +yv + -I-') 
Cn=-KK+k) 

C -- ^' 



In Tvie fern kann man das Holz etc. 

Die Constant k ist theoretisch in der That 

— 1 

— 2" 

Es war fiir jeden um die .c-Axe symmetrischen Korper 

^11 ^^3 ^12 

was auch der Fall fiir den isotropen Korper ist 

Wir fiihren neue^Coordinaten ^'r/^' ein welche so definirt werden 

wo a ^ Y wieder die Cosinusse sind. Er ist 

^^ + -^- -f- r -= ,o = ^'- + t/'-^ ^'- 

Cn = ~ = h^/^dT{rj,^' + ,9.// + n^ )F{p) 

Da in Folge der Gleichmiissigkeit der Strulctur des Korpers 
[pidzF{p)~"r/-= [/^^-^(/>)?"C" = etc = .z,. 
[fxdvFip) ^'i^ = [pdzF{p)^'h; = etc = ^ 

[lidrF{p)q" =[pdrF{p)r/' =t\.c = a. 
mithin haben wir auch 

Es ist nun 



31 



folglich 



a, = («i^ + ,9.^ ^ r/)'2i + 3'^.2( I - ^^i' - /5,* - Yx) 



0- 



D. Kitao: 



«i( I - «/ - /5/ - n") = 3^.2(, I - «i* - ;'^i' - I'l') 



folglich 



mithin Gi = 3^i2 

ob ein solches Verhaltniss herrscht ist cben so zweifelhaft wie der Einwurf 
dagegen, da jede kleine HeterogeilLit des Materials stark dieses Ver- 
hiiltniss beeinfliissen muss, und man bei jedem Versuche nie sicher sein 
kann dass, das verwendete Material homogen sei. 

Caginard de la Tour (Poggendorff Band 12 S. 518) fand in der That 
fiir einen Eisendraht 

Wertheim (Poggendorff B. yS S. 381) dagegen 

/(: = } 

F. Naumann fand allerding fiir Eisen den Werth /'^-^ ^"^ f^'' ^ndere Stoffe 
nJihrte sich I' nacli seinen Versuche dem Wertheimschen Werth. Die 
Untersuchung von Kirchhoff (Poggendorff Band 108 Seite 369.) haben 
das negative Resultat geliefert, dass zwischen den beiden Constanten 
fiir die isotropen Korper kein constantes Verhaltniss stattfinde. Cornu 
(Comptes Rendus T. 69. p. 333. 1S99) fand fiir Glas durch optisches Mittel, 
das Verhaltniss 



-\ 2/['+I ) 



zwischen 0.22 und 0.26, was den tlieoretischcn Werth 1=0.25 nahekommt. 
Mallock (Proc. Rog. Loc. V 29. 157, 1879) ^^"d fiir einen weichen Stahl 
das Verhaltniss 0.259 aber fur anderc Stoffe grossere Wcrthe. W, Voigt 
(Wiedemann 15. p. 497, 1882) fand fiir ein galvaiu'sch niedergeschlagenes 
Kupfer gcnau den Werth Y^- jcdoch kann man diese Abweichung keines- 
weges der Theorie zu Grund liegenden Hypothcse zur Last legen. Denn 
die Frage ist eben noch eine offene, in wie weit die beobachteten Abwei- 
chungen der Ungleichmiissigkcit des Gefiiges des Materials zur Last fiillt. 
Es mag dem k sein, wie cs sein wollc. Es handelt sich darum, das 



lu wie fern kaim man das Holz etc. ^^ 

Holz so zu dimensioniren dass man es als isotrop betrachten kann, dass 

als unendlich klein ansehen kann. Ich habe vierekige Stiibchen von 
Holz bilden lassen von ungefalir i cm Quadrat und von der Liinge 60 cm 
nnd dieselben der Bicgung- unterworfen, und K auf gewohnliche Weise 
bestimmt. Unterscheidet dabei eine Biegung in tangentialer Richtung, 
und eine Bieguug dazu senkrechter radialer Richtung, uud nahm an, 
dass die beiden Biegungen dieselbe Grosse K liefern miissen, wenn das 
Holz als isotrop betrachtet werden kann. 

Das Instrument, das ich angewendet habe, war wie folgt, beschaffen 
(Fig i). Eine etarke eiserne Bank DD tragt zwei scharfe Keile LL, die 
man lilngs einer Rinne in der Bank auf und niedergeschoben "werden 
kann. LL dient dem Holzstiibchen HH als Stiitzen, CC ist eim ]\Ietal- 
rahmen mit einem Keil, und hat eine Spalte, so dass man den Keil genau 
auf die Mitte des Holstiibchens legen kann. Ein Spiegel von grosser 
, Brennweite dreht sich auf dem Stifte T, und ruht auf dem Stiftchen S, 
welches das Pliittchen iM (Fig 2) beriihrt, das Pliittchen M ruht auf einen 
Biigel CC, der mittelst der Schraube 3 am dem Metallrahmen A A auf und 
niedergeschoben werden kann. Die Schneiden LL konnen mittelst der 
Sclirauben s in jeder Hohe befestigt werden. Der Metallrahmen (AA) 
steht mit dem Metallrahmen B in Verbindung, und das Holzstiibchen H 
ruht auf dem Keil k, und wird durch ein Gewicl:*" G in der Schale 
herabgezogen, und so gebogen, in dem das Holzstiibchen 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 tag i =:tacf 2a 

tacf 2a = 



_ I inn \ 



der Biegungspfeil o ist dann anniihrend 

= /. sin a 

Wo / die Liingc des Stiftchens (S) ist. Indem u auf und nieder 
schraubt kann m mit n zum Zusammenfall gebiacht werden. Die 



34 



». Kitao: 



Empfiiidliclikcit des Instrumentes ist dann selir gross, und ein '/lOOO 
Milimmeter voii deni Biegungpfeile kann leicht bestimmt werden dass 
manche Ubehtiiiide sich zeigen, wic der Einfluss der Eindriicke an dcr 
Schneide. die den Holzstaben als Stiitzen dienen, der Einfluss der Luftfeuch- 
ti^keit. 



Standort 


Altr 


Falizeit 


Breite des 
Herbstrings 


K. K. 

gebogCD tang gebogen radial 


Location 



Thuya obtusa. (Hinoki) 



Takaosan 


130 


Miirz 


0,140 cm 


14,03 X io7 


12,93 X 'o'^ 


Kern 


— 


— 


— 


0,100 cm 


13,73x107 


13,15x107 




Gifu 


160 


unbckannt 


0,030 


7,13 xio7 


672 X io7 




Kishu 


130 


Novem'oer 


0,150 
0,170 


1 1,70 X lO^ 
1 1,70 X lO^ 


ii,79x io7 
1 1,70 X io7 




»> 


60 


August 


0,140 
0,150 


14,48 X lO'' 
14,37 xio7 


14,80 X io7 
15,27 X lO^ 





Takaosan 


130 


Miirz 


0,150 


14,86x107 


i2,88xio7 


1 Kern J Splint 


» 




» 


0,050 


14,89 X io7 


13,58 XI07 


Splint 


Gifu 


160 


unbekaiint 


0,060 
0,090 


14,26 X io7 

6,56 X 107 


12,89 X I07 
6,25 X I07 


'. 



Thuya pisifera. (Sawara) 



Takac'san 


90 


December 


0,090 
0,110 


Ii,i6x io7 
10,70 X io7 


11,41 X 10'^ 

11,28x107 


Kern 


.. 


» 




0,130 


10,75 X io7 


10,99 X I07 


» 


)» 


» 




0,080 
0,080 


II.lSx I07 


10,16 X I07 


Splint 


" 


» 




0,090 


9,69 X I07 


8,70 X 107 


" 



In wie fern kann man das Holz etc. 



35 



Slandort 


Alter 


Breite des 

F.illzeit [lerbstriiigi- 


K. 

gebogeii tang 


K. 
gebogen radin' 


Locali;.'ii 



'Ihujopsis dolabrata. (Hiba) 



Kiso 



150 ! unbekannt ! 1,190 
,, i „ I 0,260 

0,200 
0,170 



10,38 X 10^ 

10,93 X lo'^ 

8,68 X io7 

9^8i X 10^ 



10,35 ^ 'o^ Kern 

11,61 X 10^ 
8.6 -| X lo'' ii Kern f^ Splint 



9,36 X lO'' 



Splint 



Sciadopytis verticilla'.a, (Koynmaki) 



Kiso 


So 


Noveml'er 


o,3co 


6,55 X TO^ 


6,06 X I o^^ 


Kern 


unbekannt 


unbekannt 


unbekannt 


0,200 


10,81 X lO^ 


10,87 K lO^ 


» 


Kiso 


80 


November 


»< 


6,58x107 


6,43 X lO^ 


Splint 


unbekannt 


unbekannt 


unljekannt 


" 


ir,i6x 10'' 


1 1,39 X lO^ 


- 



Cryptomeiia jiponica. (Sugi) 



Takaosan 


200 


November 


0,095 
0,070 


5. '5 >^ 'o'^ 
S.i/X lo^ 


5,31x107 
4,98 X lO^ 


Kern ' 


Kishu 


140 


unbekannt 


0,120 


6,35 X I07 


6,04 X lO^^ 


^ Kern g Splint 




" 


" 


0,130 


5,15 X I07 


5.6IX lO^ 


.3 Korn .3 Splint 



Tsuga Sieboldii. (Ssuga) 



Kiso 


100 


November 


1,1 80 cm 


11,35 X lo' 


1 1,59 X loT 


Kern 


" 


" 


" 


0,210 


9.33 X lo-^ 


5,64 X lO^ 


S, lint 



36 



1). Kitao: 



Standort 

1 


Alter 


Fallzeit 


Breite cles 
Herbstrings 


K, 
gebogen angent 


K. 
gebogen radial 


Location 



Zelkowa accuminata. (Keyaki) 



Takaosan 


unbekannt 


December 


0,1I2 

0,158 
0,165 


9,41 X io7 
10,11 X 10'' 
10,09 X lO'' 


9,98 X I07 

10,77 + 10^ 
10,46 X 10' 


Kern 








0,120 


9,54 X io7 


9,95 X I07 


Splint 








0,620 


4,69 X lo'^ 


5,93 X io7 


,» 








0,650 


4.53 X 10^ 


4,57x107 


» 



Castanea vulgaris var. Japanica. (Kuri) 



Takaosan 


40 


unbekannt 


0,126 
0,290 


13,21 X lO'' 
12,00 X io7 


13,36x107 
12,38x107 


Kern 


,, 


» 


„ 


0,280 


12,03 X ^°'' 


13,21 XI07 


Splint 


" 


" 


" 


0,300 


13,07 X lO^ 


15,25x107 


» 



Magnolia hypoleuca. (Ho) 



Takaosan 


unbekannt 


December 


0,230 
0,250 
0,190 


9,94 X io7 

10,62 X I07 
10,22 X I07 


1 1,36 X I07 
1 1,04 X I07 
1 1,42 X I07 


Ivcrn 


.. 


" 


» 


0,300 


10,15 X I07 


11,01 X I07 


Splint 


» 


'• 


" 


0,165 


9,85 X I07 


10,83 X lo'^ 


» 



Fagus sylvatica var. Sicboldi. (Buna) 



Takaosan 


130 


December 


0,130 


I2,l6x io7 


13,52 y 107 


i Kern J Splint 


» 


» 


., 


0,160 


11,41 XI07 


12,22 X 107 


Splint 


>» 


'• 


», 


0,230 


11,31 X I07 


12,96 X io7 


» 



I 



In wie fern kann man das Holz etc. 



37 



Standort 



Alter 



Fallzeit 



Breite des 
Herbs trings 



gebogen tangent 



K. 

DOgen radial 



Locatiun 



Quercus glauca. (Shirakashi) 



Takaosan 


unbekannt 


December 


o,i6o 


1 1,69 X lo''^ 


ii,7iXio7 


Kern 



Quercus acuta. (Akagashi) 



unbekannt 

» 


40 
40 


unbekannt 


0,500 
0,350 


13,18x107 
12,11 X lO^ 


13,36 X io7 
12,68 X I07 


Kern 

Splint 



Quercus thalassica. (Matebagashi) 



Nishigahara 


25 


September 


0,45 


I2,IOX I07 


12,19 X 'o^ 


Kern 



Fraxinus Sieboldiana. (Shioji) 



Chichibu 


48 


unbekannt 


0,276 
0,200 


15,02 X lo'^ 
14,13 X lO^ 


14,73 X lo^ 
14,41 X lO'' 


Kern 



Panlownia tomentosa. (Kiri) 



Nishigahara 


7 


September 


0,400 
0,450 


4,16 X lO^ 
4,62 X lO'' 


4,21 X lO^^ 
4,12 X lO^ 


Kern 



38 



D. Kitao: 



Pinus densiflora. (Akamatsu) 











K. 


K. 




Standort 


Alter 


Fallzeit 


Breite des 
Herbstrings 


gebogen 

auf 
tangential 


gebogen 

auf 

radial 


Location 


Mimuneya 


95 


November 


0,123 


10,36X10^ 


9,03X10'' 


Kern 


„ 


» 


,, 


0,136 


9,01 Xio^ 


9,68X107 


Splint 


Takaosan 


50 


)i 


0,1 So 
0,236 


11,86X10'' 
12,73 Xio7 


11,52X107 
12,21 X io7 


Kern 




» 


» 


0,350 


io,94Xio7 


ii,3iXio7 


" 




,. 


» 


0,198 


10,94 Xio7 


ii,59Xio7 


Splint 




,. 


>. 


0,175 


10,56x107 


10,47 X I o7 


.. 




" 


'• 


0,197 


10,73 Xio'' 


io,o8xio7 


» 



Abies umbellata. (Mom!) 



Takaosan 


160 


December 


0,200 


9o4Xio7 


9,i2Xio7 


Kcin 








0,120 
0,230 


9,33 Xio7 
10,48x107 


9,86 Xio7 
10,73 Xio7 


" 








0,170 


io,26xio7 


io,i4Xio7 


Splint 








0,260 


9,67x107 


9,89X107 


,' 








0,360 


io.i8Xio7 


9,76x107 


'• 



Icli habe 6 — 7 Male Biegungspfeile bestimmt und K fiir jeden Pfeil 
des Stiibcliens ermittelt und daraus das Mittel ^[eiiommen. Wenngleich die 
Stilbchen sich seit zwei Jahrcn in Zimmcr des Laboratoriums befanden, 
und darum vollkommcn lufttrocken sind, so war der Einfluss der Luftfeuch- 
tigkeit auf den Werth von K so gross, dass die erste Decimalstelle sich oft 
iindeit, dass die Luftfeuchtigkeit mehr die Anderung des K verursacht als 
die Breite des Herbstrings. In sofern die beidcn Werthe von K mit der 
Breite des Herbstrings in Iceincrlci ZLisaiT>:"iieaiiang stehen, und ihre 
Differenzen nur durch die Luftfeuchtigkeit veranlilsst sein konnen, konr.cn 



Ill wie feru kauii man das Hole etc. og 

vvir annchmeii class die beiden Werth von K einen und dcnselben VVerth 
zeigen konnten, wenn der Qucrschnitt dcs Stiibchens viel gcringer gewesen 
ware, als ein Quadrat i cm. Ich behalte mir vor, Stiibchen von viel 
geringeren Ouerschnitt in Untersuchung zu nehmen. 



lU'JJ.. AGRIC. CO/.f.. rOL. V. 



TA/EL II. 







/'^, 



// 



K 




Studies in the Physiological Functions of Antipodals and 
the Phenomena of Fertilization in Liliaceae. 

I. Tricyrtis hirta. 

BY 

T. Ikeda. 



With Plates III— VI. 



i 



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 b}' 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, Gramincae 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, 1S90 ami 1896 ; Osterw alder, 1S9S ; Goldflu?, 1S9S — 9. 
* Westermaier, 1S96. 



^2 T. Ikeda: 

forms {Miiscari, Hyacintlnis, Allium), I 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 Tricyrtis hirta 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, Avhen 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 Reiser'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 — 10 /x thick. 
For staining, several reagents were used : Flemming's safranin-gentian- 
violet-orange, Delafield's haematoxylin, hacmatoxylin-glycerine, Heiden- 
hain's iron-alum haematoxylin, Schaffner's anilin-safranin, his picro- 
nigrosin, Baumgarten's acid-fuchsin and mcthylene-blue. Gram's gentian- 
violet and fuchsin iodine-green mixture, l^esides these, various combin- 
ations of stains were sometimes tried. Of these, the first mentioned always 
proved to be the best, thougli the others, except Schaffner's reagents, gave 
pretty good results. 



Stndies in the Pliysiologrical Fiiiictions of Antipodals, etc. 47 



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. i). 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 is 
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, s), which is soon followed by its transverse 
division into chromosomes (Fig. 6), whose number is not exactly known. 
The microsomes which constitute each chromosome arc often clearly 
visible. Of these, those which are found at both ends of the chromosome 
gradually enlarge and this expansion of microsome granules is alwax's 
accompanied by the shortening process of tlie chromosome, until it be- 
comes condensed into a short dumb-bell-shaped rod and when the con- 
densation proceeds, the final products are nuclear 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. Ikorta: 



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. lo), 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. ii). These bodies miglit 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 Tricyrtis Jiirta there takes 
place a tetrad-division^ of the first type, as recently stated by Schnievvind- 
Thies.2 Guignard,^ in his researches on the embryo-sac formation, 
studied also Tricyrtis Jiirta ; according to him, only one, the lower of the 

> In the sense of Jutl (cf. Jucl, 1900). 
■' Sclinicwinfl-Thics, 1901. 
» C^uignard, 1882. 



studies in the Pliysiologioal Functions of Antipodals, etc. 4 - 

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 upjDer 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-sae. 

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 (PI. 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 tliis early stage is relatively deficient in 
chromatin, which is suspended on the linin-net-work, togcth.er 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 



46 T. Ikoda: 

unincreased and attached simply to the periphery of tlie 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 comj^leted. 

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 : tiieir 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. 



studies in tlie Physiological Fmictioiis of Aiitii)odals, etc. 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, 
/>.) ; 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 nucleokis (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. 

AntipodalSj Integuments, Nueellus, Chalaza, etc. 

a. Antipodal Cells. 

The antipodal cells of Tricyrtis Jiirta never change their original 
orientation in relation to the embryo-sac, as is the case with NigcUa as 
described by Westermaier. He wrote about the protrusion of the chalazal 



48 T. Ikeda: 

end of the embryo-sac of Nigella towards the underl}ing tissue as 
follows^: — " Es vertieft sich namlich der Embryosack auf einer Seite an 
der Basis der ,,Antipoden," indem Zellen des Knospenkernes resorbirt 

oder irgendwie verdriingt werden ; " So far the process accords 

perfectly with ours. In Tricyrtis Jiij-ta, howe\"er, the plane of insertion 
of the antipodal cells remains unchanged throughout the whole period of 
development, that is, they are perpendicular to the longer axis of the 
ovule and are always placed in the funnel-shaped portion of the embryo- 
sac at its chalazal end. The prolongation of the antipodal cells down- 
wards begins very early, until they attain their maximum length at the 
time of fertilization or a little later. 

Before going further I will describe here an important cytological 
feature bearing on the nutritive function of the antipodals. In the youngest 
ones, the cytoplasm is finely granular and compact in appearance. The 
small nucleus is furnished with a single, highly stainable nucleolus 
and possesses the scanty chromatin, wliich remains for a long time in 
the reticular state (Fig. 14). During development these features undergo 
a considerable change : when the chalazal end of the antipodal cells 
becomes more and more elongated down'wards their nuclei become highly 
enlarged and now begins gradually an extraordinary increase of chromatin. 
The chromatin substance becomes variously aggregated within the nucleus, 
especially along the inner periphery of the membrane : it forms a number 
of big extraordinarily dense and consequently highly stainable, usually 
angular chromatin-masscs (Fig. 18). The single nucleolus gradually be- 
comes very much smaller, so that often it is hardh^ to be distinguished 
from these chromatin-masses. 

The phenomenon of chromatin-aggregation of the nuclei constitutes 
the most remarkable fact during the development of the antipodals. What 
may be the significance of this peculiar process .^ Let us go at first to the 
literature. 

In animal as \\ell as jilant cells it has been observed by various 
authors that gland cells show the phenomena of chromatin-aggregation 

* Westcrmaier, 1890, p. 5. 



studies in the PhysioloaricJil Fniictioiis of Aiitipodals, etc. ao 

during their activity. Of secretory 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 Aconititm Napcllns, 
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 tJic clironiatin-aggrc- 
gation in the nuclei of antipodals of Tricyrtis is also the expression of their 
metabolic activity, — that therefore these orgafis play a most essential role 
in the nutrition of the embryo-sac, — that they are indeed the metabolic centre 
for 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 oi 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 b}* 
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.e. 

2 Schniewind-Thies, 1S97. ■'• Magnus, 1900. 

8 Huic, 1897-1899. " Osterwalder, 1 c. 



so 



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, 2 
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. 

b. 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 chloriodidc 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 

^ cf. Wilson, 1900, \>. 44. 
2 Wilson, I.e., p. 350. 



studies in the Plij siological Fiiiictions of Aiitipodals, etc. c i 

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 tlie 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, //., Fig. 
22 a and d). 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 parench3'-ma 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, c/i.) 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 kirgc nucleus rich 
in chromatin. 

Afterwards the nucleus of these cells becomes gradually small and 



1-2 1'» Ikp<la: 

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 witli the decay of delicate, parenchj-ma cells directly 

1 Westermaicr, I.e. 

2 Goldflus, I.e. 



studies in the Physiological Fiiiictions of Antipodals, etc. 53 

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 Avith. 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. It is 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 lionour 
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, I.e. 



54 T. Ikeda: 

Osterwalder, in his study of Aconitutn Napelbis, 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 ini- 
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 Lysichiton and Spargnniinn, observed an 
extraordinary multiplication of antipodals. whereupon he came to the 
probable conclusion that they must play an important role 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 jDrothalHal 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. Sarganf^ states quite rightly in her recent interesting paper. 

The nutritive function of the antipodals of Tricyrtis Jiirta will, I think, 
be concluded on the basis of various facts described till now. though 
necessaril}' more or less hypothetically. These facts arc : — 

1 Osterwalder, I.e. 

2 Goldflus, I.e. 

3 Balicka-Iwanowska, 1S99, p. 6S. 
•» Campbell, 1899. 

•"■ Guignard says for example as to the Lcgumiiiosae : " Les antipodes disparaissent souvent avant 
la fCcondation, par suite de la resorption du tissue nuccllaire sous-jaccnt ; d'ailleurs leur rule, encore 
assez problcmatique, parait terminc peu de temps aprCs leur formation ; dans d'autres plantes, au 
contraire, on les voit s'accroitre d'une fn(;on notable, mCme aprOs la fccondation." (1S81, p. 200.). 

8 Sargant, 1900. 



studies in the Physiolog-ical Functions uf Antipodals, etc. c r 

I.) Tlie 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 to the 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.^ 

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 role 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. 

* 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 Tncyrtis hirta, as just stated, the h'miting membrane between the nucellus 
and the inner integument is cuticularized bat the cell-zvali of the embryo-sac itself is not, so that 
though food materials from the exterior must pass generally through the antipodals, it is not impossible 
that the nuceilar cells farming the lateral side of the embryo-snc, when degenerated, are absorbed 
directly by the latter. 



56 



T. Ikeda: 



This conclusion drawn only from morphological facts, is confirmed by 
microchemical reactions, which are treated of in the next chapter. 

Microchemieal Observations. 

The microchemical reactions were made on fresh ovules, collected 
between 1-2 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- 
iiaphtol and thymol were employed for this purpose. 

After the treatment of free hand sections of ovules, chiefly longitudinal 
ones, with an alcoholic solution (20^) 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 
sufificient 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 «-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, 

» Zimmerniann, 1892. 
' Zimmcrmaiin, I.e. p. 73. 



studies in the Plijsiologieal Functions of Antipodals, etc. ey 

are carefull}'' 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 Molisch'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. 

b.) 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 Reiser'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 PI. Yl. 

In tb.e period of the archespore (Fig. 40) no starch reaction is obtained 
within the ovule. Starch grains are found in the 'wall of the ovar\', except 
in its epidermis ; they are probably the products of carbon-assimilation in 
these places themselves, 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 of 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 tlie 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 integumicnt disappears and minute dextrine granules 
appear in the micropylar region of the inner integument. 

After the union of the polar nuclei (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 



Stiulics in tlie Physiological Functions of Antipodals, etc. cq 

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 integument (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 la}'er 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 
egg-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 



6o T. Ikoda: 

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 possibl)' 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 toAvard the 
chalazal end. 

From what has been described up to this point, it will be seen tliat 
starch has never been met with in the chalaza, the antipodais, 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 w"here 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 small ness and 
scanty cytojalasmic contents of its constituents ; and water necessar}' for 
this hydrolysis may be supplied by the vascular bundle and especially by 
the spiral trachci'des, 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 smallncss and rich cytoplasmic contents of its constituents;^ w'ater 
necessary for its hydrolysis may be supplied also by spiral tracheVdes, 
which run through the centre of this group of cells. 

The chalaza and the conducting passage, which are always free from 

* Billings (1901) lias recently applied to this cell-f;i"oup the name of "nutritive tissue" 
(„l^ahrgc\vebe"). 



studios ill tho Physiological P'liuetions of Aiiti]>(><lals, etc. (3j 

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" (,,Starkestrasse ") given by him to this tissue must be 
changed into " sugar route." 

Fertilization. 

The so-called "double fertilization" takes place in Tricyrtis Iiirta. 

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 Endiinyon 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 tlie contents of the latter. 

The two generative nuclei discharged from tlie pollen-tube take their 
course to unite with the egg-nucleus and the primitive endosperm nucleus 
respectively. Thougli direct observation is Avanting, 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 tlie 
synergidae, but is always richly loaded witii 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 to a 
certain degree, so that the amount of chromatin seems to show a 

1 Guiguartl, 1899. 



^2 "f* Il^eda: 

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, e.n. and g.7i. /). 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 ^.«. .?.). 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 \\\ 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 ; anti 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 Finictioiis of Aiitipodals, etc. 6^ 

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 a and b) 
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 figiu-e 36 a corresponds 
exactly to his figure 32 (cell from the stem-apex of Psilotiiin triquetruui) , 
and also our figure 32 somewhat resembles his figure 27 (cell from the 
root-apex of Vicia Fabd) or 38 (spore-mother-cell of Eqiiisctum pabtstre). 

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 b). 

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 Zimmerman n, 1883. 

- Zimmcrmann, 1S96. p. 64. 



64 



T. Ikodii: 



surface extension for the purpose of metabolic interchanges between the 
nucleus and tlie cytoplasm, Phenomena which miglit be included in the 
same categor}' 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 Dytiscits : 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. ^ Of the vegetable cells, KohP observed in the living nuclei of 
marginal cells in the leaves oi Elodea canadensis, as well as in those of the 
hair-cells in leaves of Tradescantia 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. 39^2", b). 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 aggre- 
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 Tricyrtis differs widely from the ordinary course. For it is a 

1 See Wilson, 1900, p. 349-350. ' 

•i Kohl, 1897. 



studies in the Physiological Fnnotions of Antipodals, etc. (Je 

well-known fact that usually during- this process the cytoplasm of tlie 
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 from^the 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, ^ to take place also in. Vaillantia Jiispida belonging to the 
Rubiaceae. 

Summary. 

1. 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 cliromatin- 
masses begin gradually to dissolve away. 

4. Of the 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 tea 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 ovar}-. 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 

* Lloyd, 1899. 



(£ T. Ikeda: 

cell-c^roups near the placenta and the chalaza, then the soluble carbo- 
h}-drates 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 tiie antipodals. These 
products of degeneration may be absorbed directl}' by the antipodals or 
indirect]}' 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 Tricyrtis Jiirta 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 1 are 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 cytoplasiii and nuclei. 

10. The mode of endosperm-formation differs greatly from the ordi- 
nary one in this th.at the embryo-sac is from the beginning filled with the 
compact cytoplasm and successive nuclear divisions occur within this 
cytoplasm. 



\ 



studies in the Plij siolo^ieal Fiinctioiis of Antipodals, etc. 5- 

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. 



68 T. Ikcda: 

LIST OF PAPERS QUOTED. 

Balicka-Iwanowska, G. : Contribution a Tetude du sac cmbryonnaire chez 

certain Gamopetales. Flora 86, 1899. 
Billings, F. H. : Beitriige zur Kenntniss der Samenentwickelung. Flora 

88, 1901. 
Campbell, D. H. : Notes on the Structure of the Embryo-sac in Sparganiinit 

and LysicJuton. Bot. Gaz., 27, 1899. 

Goldflus, M. : Sur ]a structure et les fonctions de 1' assise epitheliale ct 
des antipodes chez les Composees. Journ. de Bot. 12—13, ^898 — 1899. 

Guignard, L. : Recherches sur le sac cmbryonnaire des Phaneroga 
angiospermes, Ann. d. sc. nat. Bot. VI., 13, 18S2. 

: Sur I'origine du sac cmbryonnaire ct le role des antipodes. Bull. 

de la Soc. bot. de France, 28, 1881. 

: Les decouvertes recentes sur la fecondation chez les vegetaux 



angiospermes. Volum.e jubilaire de la Soc. de Biologic. 1899. Quoted 

in Sargant, 1900. 
Hacker, V. : Praxis und Theorie der Zellcn-uncl Befruchtungslehre. Jena, 

1899. 
Huie, Lily, H. : Changes in tlic cell-organs of Droscra rotundlfolia, 

produced by feeding with (zgg albumen. Quarterly Journ. of Micros 

Sc. 39, 1897. 
Further study of cytological changes produced in Droscra. Part 

II. lb. 42, 1899. 
Juel, H. 0. : Beitriige zur Kenntniss der Tetradentheilung. Jahrb. f. wiss. 

Bot. 35, 1900. 
Kohl, F. G. : Zur Physiologic des Zcllkcrns. l^ot. Centralb., 72, 1897. 
Lloyd, F. E. : The comparative embryology of the Rubiaceac. Memoirs 

of the Torrcy Botanical Club, 8, 1899. 
Magnus, W. : Studicn iibcr die endotrophcn Mycorrhiza von Ncottia 

Nidus avis L. Jahrb. f. wiss. Bot. 35, 1900. 

Osterwalder, A. : Beitriige zur ICmbryologic von Aconituvi Napclliis L. 
P1ora. 85, 1898. 



studies ill the Pliysiolo^ical Fuuctioiis of Aiitipodals, etc. gg 

Rosenberg, 0. : Physiologisch-cytologische Untersuchungen iiber Droscra 

rotuudifolia L. Upsala, 1S99. 
Sargant, E. : Recent work on the results of fertilization in Angiosperms. 

Ann. of Bot. 14, 1900. 
Sclmiewind-Thies, J. : Beitriige zur Kenntniss der Septalnectarien. Jena, 

1897. Review by Mobius in Bot. Ccntralb., 69, p. 216 — 218. 
: Die Reduktion der Chromosomenzahl und die ihr folgenden 

Kcrntheilungen in dcii Embryosackmutterzellen der Angiospermen. 

Jena, 1901. 
Westermaier, M. : Zur Embryologic^ der Phanerogamen, insbesonderc 

iiber die sogenannten Antipoden. Nova Acta Acad. Leop. -Carol. 

57, I. 1890. 
: Zur Physiologie und Morphologie der Angiospermen-Samenknos- 

pcn. Beitr. z. Aviss. Bot. I, 2. 1896. 
Wilson, E. B. : The cell in development and inheritance. 2nd. ed. New 

York, 19CO. 
Zimmermaun, A. B. : Die Botanische Mikrotechnik. Tubingen, 1892. 
: Beitriige zur Morphologie und Physiologie der Pflanzenzelle, II, 

I. Tubingen, 1 893. 
: Die Morphologie und Physiologie des pflanzlichen Zellkernes. 



Jena, 1896. 



70 



T. Ikeda: 



EXPLANATION OF FIGURES. 



PLATE I. 

Fig. I — lo. Various stages of the first nuclear division of tlie embryo-sac-motlier-cell. Fig. 9 and 
10, Zeiss, homogeneous immersion J^ and ocular 3, all others the same obj. and oc. 4. 

Fig. II. Second nuclear division of the embryo-sac-motber-cell. SXyV- 

Fig. 12. Beginning of the embryo-sac with three degenerating sister-cells. 3X/2. 

Fig. 13. Embryo-sac with two nuclei. 2X3*2* 

Fig. 14. Embryo-sac with two polar nuclei. SXij. 

Fig. 15. Two polar nuclei concerned in copulation. 2XiV- 

Fig. 16. Mature embryo-sac. 0, ovum; syn., synergidae ; ant., antipodals. Nucleolus of primal y 
endosperm nucleus extremely large ! 4X x'a'' 

Fig. 17. Cells of the conducting passage with protein granules. rt/^A, antipodals. 3Xj\>. 

Fig. 18. One antipodal short after fertilization. Chromatin-aggregation in the nucleus ! 4Xi'2' 

Fig. tg. Two antipodals. Later stage than that of Fig. I!^'. Chromatin-aggregation less remark- 
able. Fibrillar structure in cytoplasm ! 4X1^.. 

Fig. 20. One antipodal later than in Fig. 19. Fibres in cytoplasm now becoming very distinctive. 
Leitz. 4XjV- 

PLATE TI. 

Fig. 21. Cross-section of an ovary with two ovules longitudinally cut. r//., chalaza ; /., funiculus ; 
o. w., ovarial wall ; /i/., cell-group in the placenta. 2XA. 

Fig. 22, a. The same in cross-section, showing the cell-group in the placenta. 2XD. 1>. The cell- 
group in the placenta much magnified. 

Fig. 23, Cross-section of an ovule, ax. c, axial cell-group ; n. p., parietal layer of the nucellus ; 
/. i., inner integument ; 0. i., outer integument ; ep., epidermis. Leitz 4X4. 

Fig. 24. Double fertilization, e. n., nucleus of the ovum ; g. n. i, first generative nucleus ; g. n. 2 
second;/., primary endosperm nucleus. 3X3*2. 

Fig. 25. Second generative nucleus and primary endosperm nucleus in fusion. The lerlilization of 
the ovum already accomplished. SXx"** 

Fig. 26. Secondary endosperm nucleus in spireme stage. 2 x ^.s. 

Fig. 27. a. b. The same in anaphase. Two consecutive sections. 3 x ^.s. 

Fig. 28. Two daughter-nuclei derived from the first endosperm division. 3 x j'g. 

Fig. 29. Two above daughter nuclei dividing. /. t., pollen-tube ; 0, ovum. 2 y.-^-^. 

Fig. 30. Endosperm formation. Nucleoli with pseudopodiii-likc processes. 4X3'^. 

Fig. 31. The same. Advanced stage. Nucleoli fragmenting. 

Fig. 32, The same. Advanced stage. Nucleus having peculiar shape. ^'X- \2- 

^■6' 33- The same. Nucleoli fragmenting, o, fertilized ovum. 

PLATE in. 

Fig. 34. lindospcrm-formation. Nuclei of various remarkable shapes. 4 x 3*2- 
Fig. 35. The same. Nuclei in spireme stage. 4X3''j. 



» 



studies in tho Pliysiological Functions of Antipodals, etc. 

Fig, 36. The same. Nuclear division, a. side-view ; b. polar view. 3x j^^. 

I^'g- 37- The same. Cell-walls already formed. Nuclei of remarkable forms. 4X^'j. 

Fig. 38. Embryo in almost ripe seed ; no differentiation ! 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 -i^. 

PLATE IV. 

Figs. 40—49, schematic representations of various stages of development of the ovule. 
Abbreviations : 

o. 7C'., ovarial wall ; c. ?'., external integument ; ar., archespore ; i. i. ;;/., micropylar region 
of the inner integument ; /. «., polar nucleus ; p. e. «., primary endosperm nucleus ; tin. c, 
nucellar eap ; i. ?'., inner integument; mi., nucellus ; o. e., epidermis of the ovule; cti., 
cuticularized membrane: ch, chalaza ; /., funiculus; ov., ovum; syn., synergidae; e. it., 
endosperm nucleus; ant. cfrs., disorganized antipodals; des. e., disorganized sister cells of 
the embryo-sac; r/i., raphe ; //., pollen-tube. 

Fig. 40. Archespore formation. 

Fig. 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 granule;. 



CONTENTS. 



Introductory 

Materials and Methods 

Formation of tlie Embryo-sac 

Maturation of the Embr\-o-sac 

Antipodals, Integuments, Nucellus, Chalaza etc 

a. Antipodal Cells 

h. Integuments, Funiculus and Raphe. 

c. Nucellus ar.d Chalaza 

Microchemical Observations 

a. Soluble Carbohydrates 

b. Starch and Dextrines 

Fertilization 

Formation of the Endosperm 

Summar}' 

List of Pajjcrs Quoted 

I^xplanation of Figures 



Page. 

41 
42 

43 
45 
47 
47 
50 
52 
56 
56 

57 
6r 
62 

65 
68 
70 



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I 1 Cytoplasmic contents. 



Auctor del. 



Contributions to the Study of Silk-Worms. 

I. ON THE EMBROYOLOGY OF THE SILK WORM. 



BY 



K. Toyama. 

(With Plates VII— XI.) 



Since tlie publication of Tichomiroff's beautiful work on the '' Develop- 
pement du ver a soie du murier dans I'oeuf," about ten }-ears 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. 

Methods. 

Eggs deposited on a karton arc killed by the solution (i — 2%) of 
chromic acid or by the Flemming's weak triacid solution, both heated to 
80 — 90°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 thoroughl}- washed witli water and transferred for harden- 
ing to /0% alcohol in which they remain for several weeks or even months. 



74 



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 karyokinetic 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 w'ith alum-carmine, glycerine or alum 
haematoxylln, 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 
vltellophags. 

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 Tichomirofi".* 

The closure of the blastopore takes place very slowl)'', as will be seen in 
an embryo rc])rcsented in Fig. i, PI. VII., where It appears as a wide furrow 
tapering at both ends (Fig. I, bl. a). Tliis embryo was killed at the end 



• Ticlioniiroff 1. c. l"ig. 16. 



Contributions to the Stn4l} of Silk-AVori»s. 75 

of August or September, A circular depression will be observed at the 
anterior end of the furrow (Fig. i, o). 

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 of a deep sac. A similar invagination at the primary head segment 
was also observed in a younger stage t-han 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 Avhich 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 chana"es 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 
a clear 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, i and 6, o, o^), 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. i, 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 as a 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- 



y^ Ji. Tojama: 

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 Ics premiers stades de developpement, cet entoderme 
secondaire, au moment meme de sa formation, se convertit immediatement 
en mesoderme." 

According to my observations, however, they correspond to the 
" Paracyten " of Heymons ('95) and like them gradually disintegrate into 
small 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. i), certain alterations are already found in the inner 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. Tlic 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. Tlic 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-j. They differ. 



Contributions to the Study of Silk-Worms. ^-7 

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 la3^er has already 
separated off from the ectoderm as a 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 in the median portion, its lateral arrangement becoming visible onh* 
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 tlie posterior 
opening of the blastopore, but no trunk segment is as yet to be seen 
(Fig. 12). It is in 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. 



yS 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 oi 
this stage, i.e. the resting stage. The first section (Fig. 13) passes 
through the primary head segment ; the oral cell-mass (Fig. 13, ms') 
elongates considerabl}^ 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 in a 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 iS in number, the first (the 
oral cell-mass) and the last (the anal segment) being the largest 
(compare Figs. 13 and 16, v.ith Figs. 14 and 15), This reminds us of 
Wheeler's Fig, 72, PI. 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, PI. VIII). The 
cells constituting the lateral streak are clearly of two layers, the upper 
consisting of cylindrical cells arranged regularly, while the lower laj'er 
consists of flattened cells arranged irregularly. These evidently cor- 
respond to the " paradermalcn " and the " paralecithalen Schicht " of 
rieider. 



Contributions to tlie Stinlv 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 dififere par sa forme de tous les autres. Ce segment tire son 
origine de la partie la plus profonde du sillon primitiv. Les cellules 
different egalement du reste du mesoderme : elles sent un peu plus grosses 
que les cellules mesodermiques ordinaires et leur plasma est plus clair." 
Moreover, he homologizes it with the structure which Hatschek (7"/) 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 aliens voir 
qu'en realite il n'en est rien ; cet epithelium (Middarmepithelium) a une 
origine differente." The further changes concerning the cell-mass are 
not, however, given. 

Fig. 1 8 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 at the 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. iS. In the anterior portion 
(Fig. 19), the mesoderm flattens out on both sides of the median line 



So 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 
25), and proceeds gradually to its distal end Avhich 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 arc placed latero-posteriorl}'. 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 
fuid that the disintegration of its cells becomes more vigorous with the 
ingrowth of the stomodiuni until at last the entire cell-mass complctcl}' 
disappears. This is clear!)- to be seen in Figs. 29, 31 antl 32. Fig. 29 
is a median longitudinal secticn of the [jrimar\' head segment where 



Contributions to the Study of Silk-D'orms. 3 j 

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 as in 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 invaginated cell-mass at the anterior end of the blastopore, althotigh 
it greatly resembles the entoderm-anlage of other aiithors such as Heider, 
Wheeler etc., is certainly not to be regarded as sncji in the present case. 

Then the questions naturally arise : (i) Whence arises the entoderm .'* ; 
(2) What is the oral cell-mass and the cells migrating from it ? 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 as a definite tissue. Nor do 
the mesoderm cells of the *' Vorkiefersegment " have any relation to 



•g2 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 
(a) of the tube is more elongated than its dorsal corner (c) (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 ('yy) in 
Bombyx, and erroneously considered to be the entoderm-anlage. Wlieeler 
('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. Tiic cytoplasm is very granular and has a distinctly yellow 
tint even in the unstained sections. With the elongation of the stomodial 
tube the subresopbageal bod)' 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 embrj-o talccn 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 priniar}' 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 wc 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-ventraliy. Between 
the ectoderm and the stomodial tube are seen some free colls (Figs. 46, 



Contributions to the Study of Silk- Worms. 83 

47, b.c) 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 
point, but Ave 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 H}'menoptera, though they are much 
-less distinct and more evanescent than in the lower orders of insects." 



34 K. Toyama: 

It is interesting to note here, moreover, tliat the stigma appears in 
each segment from tlie first thoracic to the eleventli. In the meso-and 
metathoracic segments we are able to observe faint depressions of the 
ectoderm, which may represent the rudimcPits 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 
stio-mate du metathorax ne disparait pas. il demeure sous la forme d'um 
stio-mate rudimentaire avec son faiscean correspondant tracheal, meme 
chez la larve adulti ; ce stigmate rudimentaire se trouve fortement avance 
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, where a 
stigmata is provided with a closing bow, a closing band, and a closing 
lever, as is described by Krancher ('Si) 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 is a 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 th.in (gl.), constituting the " Grenzlamelle " of Heymons. 
The " vordcrc ICpithcllamcllc " has developed more than before and 
close to its ventral side the subccsophageal 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. 



Coutributioiis to the Study of Silk-Worms. S5 

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 suboesophageal 
body. But the development of the entoderm-anlage will become clear 
when we come to examine the serial sections mentioned below. 

Fig. 55 is a longitudinal section through the primary head segment 
of a slightly more advanced stage, showing the elongation of the " vordere 
Epithellamelle." Series of transverse sections of the head segments of 
this stage are given in Figs. 56 — 62,, 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 suboesophageal body is to be seen. In this 
flat cell-mass we again observe the thick lateral portions (ent) which 
have been already referred to. In the 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 suboesophageal 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 suboesophageal body 
is 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 



g^ K. Toyama: 

3. 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 6t,, sm. ms) the somatic layer, but a closed coelomic cavity 
is not formed. On the dorsal and the inner portions of the splanchnic 
part of the curved end of the mesoderm will be seen the entoderm-anlage 
or the 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. In the 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 trabecules de cellules vitellines " ; namely 
a middle and two laterals, and is of the opinion that the neurilemma and 
the fat bodies arc derived from the former, while the epithelium of the 
mid-gut comes from the latter. On this point ho says; " ils donnent 
naissancc a I'epithclium de I'intestinc moyen, epithelium que se constitue 
des cellules de rentoderme secondaire, emanant de ces trabecules des 
cellules." We have also frequently observed such cells corresponding^ 
in all particulars to those given in his Fig. 54 Bp. (sec our Figs. 59, 64, 
and 91 l^c), but we have not been able to find any such cells in any 
stage of transition into the entoderm. On the contrary, we frequentl>'^ 



Contributions to the Study of Silk- Worms. g^ 

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 is a frontal section of the posterior portion of the stomodium 
of a more advanced embryo. We observe in it the karyokinetic division 
of the nuclei of the epithelium of the mid-gut, by which the cells of this 
region arc multiplied thus causing its closure. 

From the foregoing accounts, we may sefely conclude that tJie afiterior 
entoderm-anlage is formed by the proliferation of the epithelial cells of the 
stomoditivi, and is consequently ectodermal in its nature. Neither the 
mesodermal cells formed by the cells of the blastoporoiis invagination^ nor 
the yolk cells ^ have anytJiing to do with iJie formation of this struct7ire. 



The proetodium and the Malpighian vessels. 

The first appearance of the proetodium 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 proetodium is seen only as a 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 proetodium 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 presentent en effet que comme de 
simples excroissances tubulaires de I'intestin posterieur. Comme on sait 
une larve adulte posside six vaisscaux malpighians, qui debouchent dans 
I'intestin posterieur par deux conduits communs. Ce sont ces conduits 



33 K. Tovama: 

qui apparaissent des le commencement comme de simples excroissances 
de I'intestin posterieur, se partagent ensuite chacum en trois tubes comme 
les trones tracheaux principaux emanant des stigmates, se divisent en 
rameaux secondaires." Hatschek i^'j'j) also observed three tubules on 
each side of the proctodium or hind-gut of Bombyx, and says that "die 
drei malpighi'schen Driisen jeder Seite miinden durch ein gemeinschaft- 
Hches Anfangsstiick in das blinde Ende des Hinterdarmes." According 
to our observation, tliese arise as three separate pairs of hollow outgrowths 
from the beginning as will be seen in Fig. ^6, 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 dif^cult. Fig. 74 is a sagittal section of the 
proctodium of a 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 siiown 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. 
We are 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'ctre familiarise plus tard 



Contributious to the Study of Silk-Worms. go 

avec le role important que remplit rentoderme secondaire dans la formation 
de different organes, j'incline a admettre que la partie essenticlle des 
organes sexuels se constitue aussi a ses depens." But they originate from 
the mesoderm as the former facts show. 

Summary : i. The posterioj' etodenn-anlage is derived first from 
the epithelial zvall of the proctodium in the saute way as the anterior 
entoderui-anlage is derived from that of the stomodium. 2. Malpigiiian 
vessels arise as three separate pairs of outgroivths from the blind end 
of the stoniodinm. 3. Genital cells differentiate from the cells of the 
somatic mesoblast. 



II. The vitellophags and other cellular elements 
found in the yolk. 

a. Vitellophags. 

Just as in the eggs of many other insects thus far studied, we here 
also find in the )'olk various cellular elem.ents among which the most 
conspicuous are the vitellophags, or cells left in the yolk at the time when 
the other cleavage cells are traveling towards the surface of the &^^ 
to form the blastoderm. In certain cases, however, all the segmentation- 
nuclei come to the surface of the &ZZ^ ^"^1 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, Ave find a single vitellophag in the centre of the yolk. 
This multiplies by direct division (Fig. 6Z) when the embryo attains 
the stage given in Fig. i, so that in more advanced stages we see many 
nuclei in the centre of each of the yolk-segments (Fig. g,j'.b). 

The cytological nature of these vitellophags is as follows : — The body 
of the cell is large as compared with other cellular elements found in 
the yolk. It has a large round nucleus with fine granular chromatin 



90 K. Toyama: 

scattered in it. 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, 
32, 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. 6j 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^*, sS 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 Deimaptera 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. 

c. 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 



Coiitribntions to the Study of Silk- Worms. n j 

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. 6^, 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. 6"]. 

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 &gg, 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 small 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, 49 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 a more advanced stage in which the subcesophageal body is well 



02 K. Toyaiua: 

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. 

In a 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 c}'toplasm 



Contributions to the Study of Silk- Worms. g? 

(Figs. 13 a, 66 d.c). These granules gradually increase in number and 
with this the nucleus dissolves, till at last the cells 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 i. from the migratory cells of the ectoderm ; 2. from those of the 
mesoderm ; or 3. from the free cells of the oral cell-mass. 

I. That they arise out of the migratory cells of the ectoderm is to 
be seen when we compare Figs. 2^ 5^ with Figs. 13, 19, 22 par. In 
Figs. 2,'' 5"^ 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 seen in 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 embr}-o, such as the proctodium, stomodium etc. 

From the foregoing statement, wc are able to say that there arc four 
sorts of migrating cells in the yolk. The first are the vitellophags derived 
from the remainder of the segmentation mic.lei. 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 prodticed by the disintegration of the oral cell-mass. These also 
undergo des'eneration. 



III. The endoskeleton of the head, with reference to the 
salivery gland and a new ghand. 

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. 'jj') 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. yj 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 in the next segment at the 
outer base of the second maxillae (Fig. 80 n). Another pair of invagina- 
tions is also to be seen at the inner base of the second maxillae (Fig. 
80 slg). The latter arc the origin of the silk glands. Figs. 81 — 82 are the 
consecutive scries 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 



Fig. I. 



tenti 




coalesce with one another and the b'mit of each segment is no more 

visible (woodcut Fig. 1). 

tent^~--, first and second tentorium ; 
ex. md, attachment of extensor mandi- 
bulae ; fl. md, attachment of flexor 
mandibulae. lb, labrum ; an, anten- 
nae ; mx^~--, first and second maxillae ; 
si. g, silk gland ; n, new gland or hypo- 
stigmatic gland ; th. P""'-, first-third 
thoracic legs ; H-shaped dotted line, 
tentorium aniage within head. 



sl.s. 



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. Jj 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 and the 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 
Fig. I, dotted line). 

These tubes are the aniage 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 



q6 ' K. Toyama: 

the posterior inner end of the mandible \vc again meet with a large 
invagination (Fig. 86 fi. md, and Avoodcut 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. Sy, 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 it 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. I). In the transverse section we observe these 
cell-masses on both sides of the suboesophageal body in the prothorax 
(Fig. 90 n), while in the sagittal section the first portion of them appears 
as an invaginated tube (Fig. 92 n). Fig. 94 represents a frontal section 
through the head and the 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 trilobcd, resembling fat-tissues ii> 



I 



Contributions to the Study of Silk-Worms^ 07 

appearance, and is situated on the ventral side of the first stigma to which 
it is 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. In the 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 length is now about 16 mm. 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), Avhile the cells of the fat tissue are 
small, their nuclei circular and 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 oenocytes 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 give a short summary of the above statements it will be 
as follows : 

7. 1)1 the Diaudibnlar segment, three pairs of invaginations take 
place ; the most anterior {beizueen the labmui und the mandible) becomes 
the first tentorium, the second pair gives rise to the seat of the extensor 
jna7idibnlae, zvhile the last becomes the flexor manhibnlac and salivary 
gland. 

2. In the first maxillary segments, there is a pair of invaginations 
ivJiich become the second tentorium. 

J. In the second maxillary segment, lue again meet roith tivo pairs 
of itivaginations, the inner of which forms the silk gland, zchile the lateral 
ones groiv into a gland which is situated on the inner side of the first 
stigma in the larva, and which zee zvill call the hypostigmatic gland. 



gS 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 Lcpidoptera, Kowalevsky ('71) v/as 
the first naturalist who clearly described the formation of the niesoderm 
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 10 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, liinglichen 
Rinne auf, deren Eodenzellen, 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 I'oeuf " describes and figures 
the formation of the germ layers. Concerning the mesoderm formation, 
he maintains the opinion that " elles procedent, avant tout, de I'ectoderme 
et 2) de I'cntoderme." 

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 ('Sy) observation on Thyridoptcryx also shows the inward 
growth of cells from the primitive furrow to form the mesoderm, and 



f 



Contributions to the Study of Silk-Worms. gg 

Ill's Fig-. VII., PI. I., corresponds exactly witli our Fig. 2 on this point. 
He says : " The inner layer is not strictly invaginated, for it is 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 otlier 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 Scheiria 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 
Korperregionen desselben Embryo verschiedene Form der Mesoderm- 
bildungen vor." 

The results obtained by my investigation on silk-worms as above given 
quite confirm the opiriion of the last named author. The deep invagination 
in the anterior end of the blastopore closely corresponds to a similar 
groove on the cephalic lobe described by Bruce and figured in his Fig. 
VIII. PI. I., while the inward growth of cells from the bottom of the me- 
dian furrow in the 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 Kowalevsk\-. 



lOO K. Toyania: 



The entoderm. 



In tlie interpretation of the insect-gastrula the entoderm has always 
played an important role. 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 }'et 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 in a masterly manner by Heymons ('95) and Schwartze ('98) we 
shall confine our remarks mainly to the Lepidoptera. 

In the Lepidoptera, Tichomiroff was the first wlio minutely described 
the entoderm formation. In his first paper ('79), he considers that it arises 
from the ento-mesoderm, but in later papers ('Si, '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 him in the following words : "a portion of 
the inner la5'er on each side of the embryo becomes separated from the 
other parts of the inner la}-er. These portior.s 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 similarlj- 
mistaken in his study of Chironomus. 

Graber ('90) made many valuable observations on the bipolar origin 
of the entoderm in Bombyx, Pieris, Gastropacha etc. Alth.ough 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 tlie )-olk-cells. Comparing 
other insects, he says ; " dabei nehmcn wir vorliiufig an, dass der vordere 
und der hintere Driisenblattkcim aus dem I'tychoblast und nicht aus dem 



f 



Coutributious to the Study of Silk-Worms. i o i 

Ectoderm des Stomo-und Proctodiums liervorgelit." Hut according to 
his Fig. III. Taf. X., which closel}' corresponds to our Fig. 54 or 
55, we might say that the entoderm arises out of the epithelium of 
tlie stomodium. 

Schwartze ('9S) made a very valuable observation on this [)oint 
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 
aufeinander zuwachsen, bis sie sich jederseits in der Mitte treffen, und 
sich dann in F'olge starken Breitenwaclisthums erst ventral, dann dorsal 
in der Mediane vereinigen. Der Mitteldarm ist also, abgesehen 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 Tichomiroffand 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 " i. ein Paar 
von seitlichen Divertikeln, deren Hohlraum durch seitliches Auswachsen 
des Urdarmlumens hervorgegangen ist und 2. eine mediane an die paarigen 
Divertilscln sich dicht ausschliessende Zellmasse. Aus den Divertikeln 
werben die Mesodermmassen des Kopfsegments und des spateren Mandi- 
dularsegmcnts, wilhrend 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 



I02 K. Tojama: 

Abschnittes rein mesodermatische Bildungen, beziehungsweise Anlageii, 
wiihrend die gennaiiten zwei Segmeiite gemischter Natur sind, dass heisst 
ausser dem audi ihnen zukommenden und sogar selir stark eiitwickelten 
Mesodermantheil zugleich die Entodermaiilage enthalten." 

Voeltzkow ('89), on tlie other hand, observed quite correctly tlie 
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 : 
"Audi ich kann mich mit Kowalevsky und Meider's Darstellung nicht 
einverstanden erkliiren und stimme vielmehr mit Voeltzkow und Lecaillon 
iiberein. Beide Forscher finden, dass bei Melolontha bezw. einer Anzahl 
untersuchter Chr}'someliden der Ursprung des Mitteldarmepithels in zwei 
vorderen und zwei hinteren, vom Vorder- bezw. vom Enddarm auswach- 
senden ventrolateralen ektodermalen Lamellen zu suclien ist, die durch 
ilire Vereinigung in der Mitte, sowie in der ventralen und spiiter in der 
dorsalen Medianlinie das Mitteldarmrohr entstehen lassen. Das Wachs- 
thum der ektodermalen Lamellen geschieht iiberall ohne Beziehung der 
Zellen des unteren Blattes, so dass an dem ektodermalen Charakter das 
gesammten Mitteldarmepithels kein Zweifel herschen kann." 

Comparing now the \'arious 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 sam.e 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 yy 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 



rontributions to the Study of Silk- Worms. 1 03 

same way as the migration of the cells takes place out of the oral 
cell-m.ass in silk-worm. Wheeler says : '' Be this as it may, in later 
stages I believe it can be shown that cells do migrate into tlie yolk from 
the embryo and especially from the entoderm-centrcs. 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 
Mesoderme et de I'epithelium de I'intestin moyen est tres 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 oth.cr orders of insects, the formation of the entoderm has been 
mostly studied in Orthoptera. Ayers ('84, in Occanthus) 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, '91, 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 

* The cells here are 1 think undoubtedly the oral cell-mass. 

* If \ve look at his Figs. 32, 53, and 34, and the description that " median portion thus proliferated 
beyond limits of the ectoderm is the anterior or oral entoderm-ccntrc," it is clear that in Xyphidium 
the entoderm arises from the ectoderm by cell-proliferation. 



I 



104 ^* To J a ma: 

Darmtraktus ausschliesslich ektodernialer 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 Kow.ilevsky {'S6) 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 Voelotzkow, 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 
hochst eigenthiimlichc, bei anderen Insekten bisher vollig unbekannte 
Verhilltnisse nachgewiesen, nach dcnen es mindestens moglich ist, dass 
hicr die Driisenblattkcime zum Theile aus eanz selbstiindigen Einstiil- 
pungen des Keimstreifenepitliels 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, IIe)-mons ('95) expressed his view 
in the following words : " Nach den bisherigen Untersuchungen zu 
urtheileii, ist es daher in hohen Grade unwahrscheinlich, dass das 
Mittledarmci)ithel der Musciden aus dem untercn Blatt (Entomesoderm) 
entsteht." 

Quite recently ICscherich ('00, '01) has found tlirec folds in the anterior 
portion of the blastopore nf Musca, of which he writes "sic stellen einen 
Theil der Mesodermanlage dar ; die mediane Falte dagegen ist, wie wir 
gleich sehen werdcn, die vordere Anlage dcs Entotierms. W'ir haben also 
in diesem Stadium bci den Musciden in der That ganz iihiiliche Verhaltnisse, 
wie bei Sagitta, worauf ja bekannth'ch sclion JUitschli und Kowalevsky 
hingewiesen haben." Ami he finally came to the CDUclusion, " dass I. bei 
den Musciden sich sehr friihzeitig eiii J'!ntodcrm anlegt, dass 2. dieses 



I 



Contributions to tlie Study of Silk-AVorms. 105 

Entoderm ein Abkomniliiig des Blastoderms ist und dass 3. die Differeiizie- 
rung der beiden primaren Keimbliitter durch eine typischc 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 frop.i 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. 
Carriere ('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 
ICntoderm und Mesoderm gleichzeitig auf dem Wege der Theilung der 
Zcllen des Blastoderms und ihrer PLinwanderung." 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 and the anal. But since the entoderm of many other 
animals arises from a .'^ingle centre it is tacith' assumed that such must 
originally have been the case also with insects, and that the present 
bipolar condition must be due to a secondar}' modification. Starting with 
this postulate, Kowalevsky has formulated his hypothesis that " bei der 
so in die Pange gezogcnen Gastrula der Insekten der mittlere, das 
PvUtoderm lieferndc sack so ausgezogen ist, das er in der ^Nlittc ganz 
vcrschwindet und nur an seinem vorderen und hinteren Endc bestehcn 



io6 



K. Toyama: 



bleibt." Rut if as we assume that the entoderm of Kowalevsky corresponds 
to our oral cell-mass whicli 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 lost in insects, 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 b)^ Dohrn ('96) to have some 
relation to the yolk cells. Avers ('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 Leibeshohle." Cholodkowsky ('91) also holds the same opinion 
respecting Phyllodromia. 

Korotnefif ('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- 
korperchen mesodermaler Abkunft. Sic entstehen aus Mesodermzellen, 
welchc nicht bei der Bildung der Ursegmente sich betheiligt hatten, 
sondcrn zwischcn diesen in der Medianlinie des Korpers ihren Platz 
beibehalten." A similar view is held by Lccaillon ('98) in the case of 
Chrysomelidac, and by Schwartzc in the case of Lasiocampa. But in the 
case of Lasiocampa, the last author states that " diesc Einwandcrung 



Contributious to the Study of Silk* Worms. 107 

beschriinkt ist auf diejenige Stelle des Embryo?, an die sich die Ektoderm- 
rinne von zuletzt schiiesst, d.li. auf einen sehr geringen Theil dcr 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 whicli forms the blood cells. 

From all the opinions above considered those of Korotneff and 
Heymons are in 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, 1901. 

Zoological Institute, College of Agriculture, 
Tokyo Imperial University. 



io8 



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, 18S4. 
Bobretzky, N. {'/S), Uber die Bildun^^ des Blastoderms und der Keim'bliitter 

bei den Insekten. Zeitschr. wiss. Zoologie. Bd. 31, 1878. 

Boas, J. ('99), Einige Bemerkungen iiber die Metamorphose der Insekten. 
Zool. Jahrf. Abth. f. Syst. Geogr. and Biol. d. Thiere. Bd. XII. 1899. 

Bruce, A. T. i'Sy), 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 P^ntwickelung der Mauerbiene (Chalicodoma muraria) 

im Ei. Arch. f. mik. Anatomic, Bd. 35, 1890. 

('90). Die DriJsen am crsten Hinterleibsringe der Insektenembryo- 



nen. Biol. Centralblatt, 1891. 

Cholodkowsky, N. {'S8), Uber die Bildung des Entoderms bei Blatia germa- 
nica. Zool. Anzeiger, Jahrg. 11, 1888. 

{'go), Zur Embryologie von Blatta germanica. Zool. Anzeiger, 

Jahrg. 13, 1890. 

"('91). Die Embryonalentwicklung von rh)'llodromia germanica. 



Mem. Acad. St. Pctersbourg. Tom. 38, 1891. 

Deegener, P. ('00), Entwicklung der Mundwerkzeuge und des Darmkanals 
von Hydrophilus. Zeitschr. f. wiss. Zoologie, Bd. C8, 19CO. 

Dohrn, A. {'76), Notizen zur Kenntniss der Insektcncntwicklung. Zeitschr. 
f wiss. Zoologie, Bd. 26, 1876. 

i'oi), Entwicklung der Mundwerkzeuge und des Darmkanals von 

Hydrophilus. Biol. Centralblatt, Bd. XXI. 1901. 

Escherich, K. ('ooj, Uber die Bildung der Keimbliitter bei den Musciden. 
Nov. Acta. Leop. Carol. Bd. LXXVII., 1900. 

('01), Das Inscktcn-Entoder.m. Biol. Centralblatt, Bd, XXI., 1901. 

Graber, V. ('88), Vergleichcndc Studien iiber die Keimhiillen und die 



Contribntions to the Study of Silk-Worms. jqq 

Riickenbildung- der Insekten. Denkschr. Acad. VViss. Wien, Bd. 
14, 1S88. 

— *('89), Vcrq^leicheiule Studicn iiber Die Embr^'ologie der Insekten 
und insbesondere der Musciden. Denkschr. Acad. Wiss. Wien, 
Bd. 56, 18S9. 

— i'90), Vergleichende Studien am Keimstreifen der Insekten. 
Denkschr. Acad. Wiss. Wien, Bd. 57, 1890. 

— ('90» Bemerkungen zu J. Carrierc's Aufstaz ,, Die Driisen am ersten 
Hinterleibsringe." Biol. Centralblatt. 1891. 

— *('9i). Bcitrage zur vergleichende Embryologie der Insekten. 
Denkschr. Acad. Wiss, Wien, Bd. 5«, 1S91. 

— ('91), Uber die embryonale Anlage des Blut-und Fettgewebe der 



Insekten. Biol. Centralbhatt, Bd. 11. 

Hatschek, B. {I77), Beitriige zur Entwicklungsgeschichte der Lepidop- 
teren. Jenaische Zeitsclir, f. Naturw., Bd. 11, 1877. 

Heider, K. ('85), IJber 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 Insektenkorpers, aus dem 
Anhang z. d. Abhandlungen der Konigl. preuss. Akademie d. Wiss. 
z. Berlin, 1895. 

('95). Die Embryonalentwicklung von Dermapteren und Ortho- 

pteren, unter besonderer Beriicksichtigung der Keimbliitterbildung, 
Jena, 1895. 

('95)5 Grundziige der Kntwicklung und des Korperbaues von 



Odonaten und Epheineriden, aus dem Anhang zu den Abhandlungen 
d. Konigl. preuss. Akademie d. Wiss. z. Berlin, 1896. 

Kowalevsky, A. ('71), P!^mbryologische Studien an Wiirmern und Arthropo- 
den. Mem. Acad. imp. scienc. St. Petersbourg, Scr. 7, Tom. 16, 
No. 12, 1871. 

('86), Zur embryonalen Entwicklung der Musciden. Biol. Central- 
blatt, 6 Bd. 1886, 



no K. Toyama: 

Korotneff, A. ('85), Die Embryologie dcr Gryllotalpa. Zeitschr. f. vviss, 
Zoologie, Bd. 41, 1S85. 

('94). 2ur Entwickluiig' des Mitteldarmes bei den Arthropoden. 

Bio! Centralblatt, Bd. XIV., 1S94. 

Koschevnikov, G. A. ('00), Ueber den Fettkorper und die Oenocyten der 

Honigbiene. Zool. Anzg., 1900. 
Krancher, 0. ('81), Der Bau der Stigmcn bei den Insekten. Zeirschr. f. 

wiss. Zoologie. Bd. XXV., 1S81. 
Kulag'in, (98), Beitriige zur Kenntni.s.s der Entuicklungsgeschichte von 

Platygaster. Zeitschr. f. wiss. Zoologie, Bd. LXIII., 1898. 
Korschelt und Heider, ('92), Lehrbuch der vergleichenden Entwicklungs- 

geschiclite der wirbelloscn Thiere. Hefr, 2, Jena, 1892. 
Lecaiilon ('98), Recherches sur Toeufet sur le developpement embryonaire 

de quelques Chrysomelides. Pairs, 1898. 
Mayer, P. {!/(!>), Uber Ontogenie und Phylogenie der Insekten. Jen. 

Zeischer. f. Naturvviss. 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, ('9S), SuH'origine dell'intestins medio nella Mantis religiosa. 

Palerms, 1898. 
Schwartze, ('99), Zur Kcnntniss der Darmentwicklung bei Lepidopteren. 

Zeitschr. f. wiss. Zoologie. Bd. 65, 1899. 
TichomirofF, A. ('91), Development du ver a soie du murier dans I'oeuf. 

Rapport presente a la chambre de commerce de Lyon. 1S91. 

('79). Uber die Entwicklungsgeschichte des Seidenwurms. Zool. 

Anzeiger, Jahrg. 2, Nr. 20. 

('92), Aus der Entwicklungesgeschichte der Insekten. Festschrift 



zum 70, Geburtstagc Rudolf Leuckarts. Leipzig, 1892. 



Contributions to tlie Study of Silk-Worms. 1 1 1 

Ticbomirowa, 0. ('50), Zur Embryologie von Chrysopa. Biol. Centralblatt, 

Kd. 10, 1890. 
Uzel, H. {'97), Vorlaufige Mittheilung iiber die Entwicklung der Thysa- 

nuren. Biol. Centralblatt, Bd. XX., 1897. 
Will, L. ('38), Entwicklungsgeschiclitc der viviparen Aphiden. Zoo!. 

Jahrb. Abth. f. Anatomic und Ontogenie, Bd. 3. 1888. 
Witlaczil, E. ('84), Entwicklungsf^cschichte der Aphiden, Zeitsclir. f. 

wiss. Zoologie, Bd. 40, 1884. 
Wheeler, W. M. ('89), The embryology of Blatta germanica and Doryphora 

decemh'neata. Jour, of Morphology, vol. 3, 1889. 
('93). A contribution to Insect Embryology. Jour, of Morphology, 

Vol. 8, 1S93. 
Voeltzkow, A. ('89),* Entwickhmg im Ei von Musca vomitoria. Arbeit. 

Zool. Zoot. Inst. Wiirzburg, Bd. 9, 1889. 
('S9),'" Melolontha vulgaris. Ein Beitrag zur Entwicklung im Ei 

bei Insekten. Arbeit. Zool. Zoot, Inst. Wiirzburg, Bd. 9, 1889. 
Schaflfer, C. ('89), Beitrage zur Histologic der Insekten. Zool. Jahrb., 

Abth. f. Anatomic u. Ontogenie, Bd. 3, 1889. 
Verson, E. Vison, ('91), Cellule glandular! ipostigmatische. Padova. 1891. 
Wielowiejski, H. v. ('86), Uber das Blutgewebe der Insekten. Zeischr. f. 

wiss. Zoologie, 1886. 
Selvatico ('82), Sullo soilippo embrionale dei Bombicini. Annuarid del la 

r. stazionc bacologica di Padova. 1 882. 

* The asterisk marks the cases in which I have not been able to gain access to the original 
paper. 



112 K. Toyaina: 



Explanation of Plate I. 

Fig. I. Suif.ice view of embryo with blastopore not yet closed (^taken from an egg, one month 
old : 28th, August.) Zeiss A x 4. o, anterior widening of blastopore ; pel, 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 Dx 4. 
Fig. 2. Section through primary head segment ; knife passing through anterior portion of blasto- 
porous widening (Fig. I. o). 

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.* Section of anal segment. 

Letters as in preceding figures. 
Fig. 2." Transverse section through anterior wiilening of blastopore already closed (taken from an 
embryo somewhat older than Fig. I). 

am, amnion ; ect, ectoderm ; o, ectodermal depression, jar, " paracyfen," 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 blastojiore about to close. At (i) it remains open (taken from 
the egg on November 30th.) Zeiss A X4. 

o*, ectodermal depression formed after closure of anterior widening (Fig. L o) "f 
blastopore. 
P'igs. 7— II. Cross sections taken from embryo as above figured. Zeiss Dx4. 

Fig. 7. Section through ectodermal depression (Fig. 6. o") at juimary head segment, am , aniniion ; 
ect, ectoderm ; o', ectodermal dej)ression ; m'^, mesoderm ; ms*, cell-mass sprung from 
invaginated gastrula or oral cell-mass ; ms', detached cell tmm oral cell-mass, remaining 
letters as in the preceding figures. 
Fig. 8. Section through middle portion of embryo. 

Lettering as in jircceding figures. 
Fig. 9, Section through a portion, little posterior to the iirccc'ling figure. 

Lettering as in preceding figures. 
I'ig. 10. Section through blastopore at Fig. 6. a., showing later.d overgrowth of cctodi-rm to form 
mesoderm or inner layer. 



I 



Contributions to the Study of Silk-Worm. j j o 

Fig. II. Section of anal segment. 

I^ettering as in preceding figures. 
'Fig. 7." Portion of same section, highly magnified, showing two migratory cells from oral cell-mass. 
(Zeifs Ap. o. 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 Ax 4.) 

p.cl, procephalic lobe ; cd, anal segment. 

Figs. 13 — 16, cioss 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; o*, 
ectodermal depression at primary head segment ; ms*, oral cell-mass ; ms', detached cells 
from the oral cell-mass ; d.c, degenerating cells ; vit, vitellophags. 
Fig. 13.'' Portion of same section more highly magnified, sliowing degenerating cells and vitellophags. 
(Zeiss Ap. o. 2 mm x Comp. 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 

pireceding. 
Fig. 17. Median longitudinal section of embryo, taken out on March i8th. (Zeiss DX4.) 

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.c*, migrating mesodermal cells; par., "paracylen" 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 Dx5.) 

am, amnion; n.f, neural furrow; ect, ectoderm; ms", " paradernialen .'^chicht "'; ms'' 

" paralecithalen Schicht"; vit, vitellojihags. 
Fig. 27. Transverse section through procephalic lobe, showing migratory colls from oral cell-mass. 



I j^ K. Toyama: 

am, amnion ; ect, ectoderm; nis, mesoderm; ms', oral Cell-miss ; ms-, delaclicd cells 
from oral cell-mass. 
Fig. 28. rortion 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.) 

nis', oral cell-mass ; ms2, detached cells from oral cell-mass ; yl, yolk granule?. 
Fig. 2g. Median longitudinal section of embryo at same stage as that represented in F'ig. 18, which 
shows faint depression of stomodium at st. 

A, anterior ; F, posterior direction ; st, stomodium ; ms, mesoderm ; ms^, oral cell-mass ; 
ms-, detached cells from oral cell-mass ; dc, degenerating cells ; vit, vitellopliags. 
I*'K- 30. Surface view of emljryo taken out on March 27th. 

lb, labrum ; at, antenna; md, mandible; mxi"^, first and second maxillae; th. l^'^*; 
first, second and third tlioracic legs; n.f, neural furrow ; cd, anal segment. (Zeiss A x 4.) 
Fig. 31. Sagittal section of same embryo as that represented in the above, slowing 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. Here 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 tlirough the median 
longitudinal line. (Zeiss Dx 4.) 

st, stomodium ; ms, mesoderm ; ms-', detached cells from oral cell-mass ; a, elongation 
of lateral portion of stomodium, or " vordcre EpithcUamelle," ms", anlage of suboesophageal 
body, from the anterior portion of which (a) blood cells are formed. 
A, anterior ; P, posterior direction. 
P'igs. 41—50. See explanation of Plate in. 

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 Dx 4.) 

ect, ectoderm; ms, mcsoilerm ; d.c, degenerating cells; st, stomoilium ; a, lateral 
prolongation of stomo<Hal wall to form mid-gut ; b.c, blood cells. Other letters as 
in i)rcceding. 

Fig. 51. Surface view of shortened embryo, taken from egg in April (Zeiss A x 4.) 

lb, labrum ; at, antenna ; md, mandible; mx.**"-), first and second maxilla: th. ].<'"*\ 
first, second and third thoracic legs ; sl.g, opening of silk gland; ab. 1(*"«°), abdominal 
legs ; stg, stigma ; n.f, neural furrow. (Zeiss Ax 4.) 



rontribiitions to the Study of Silk-Worms. 



115 



Fie;. 52. Transverse section tlirougli antenna! region of same embryo as that rc|>rc--ente(l in Tig. 5 
showing cntoderm-anlage. 

ent," entoderm anlage or lateral prolongation of btcmodial wall ; n, nerve. Other lettei s 
as in preceding figure?. 
Fig. 53. Sagittal section through primary head segment of same embryo as in preceling figure, she a - 
ing stomodial tube and entodcrm-anlage. 

cpl. V, '' vordere Epithellamelle "; st, stomodium ; ms, mesoderm ; g.l, " Grenzlame'la "; 
s.b, suboesophageal body ; b.c, blood cells. 
Fig. 54. Portion of same series of above section more highly m:ignified, showing blood cells. (Zeiss, 
A]\ 2. mmxComp. 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. c;5. Sagittal section of emljryo, somewhat more advanced than thit represented in Fig. 52 (taken 
from egg on April, 4th. Zeiss Dx4). 

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, 

Dx4. 

r'igs. 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 ; n.c, 
nerve cord; md, mandible; ms*, first maxilla; n.f, neural furrow ; ms, mesoderm; 
sp. ms, splanchnic mesoderm ; sm. ms, somitic mesoderm ; cce, coelomic cavity ; tent', 
second tentorium ; d.c, degenerating cells; th. 1^, second thoracic leg; ce, oenocytes. 
Fig. 64. Transverse section through sixth abdominal segment, same embryo as above. (Zeiss 
D X 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 its epithelial cells. 

epl. v, "vordere Epithellamelle"; g.l, "Grenzlamelle": sp. ms, splanchnic mesoderm; 
st, stomodium. (Zeiss, Ap. 2. mmxoc. 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 jMecediDg figure (magnification 
as above). 

d.c, degenerating cells ; ms^, immigrating cells from oral cell-mass : vit, vitellophags ; 
yl, yolk granules. 



Il6 K. Toyaina: 

Fig. 68. Volk Ijall. tnkcii from embryo as rcpresentccl in Fig. I, sliowing direct division of nucleus. 
Fig. 70. Anal segment of embryo as represented in Fig. 30. (Zeiss, 73x2). 

n.f, neural furrow ; a, proctodial depression. 
I''g- 73- Tiansverse section through anal segment of embryo as represented in P'ig. 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- 
74. (Zeiss, Dx4). 

pt, proctodium ; m.v, m;\li)igliian vessel; ms, mesodeim; n.c, nerve cord. 

Explanation of Plate X. 

Fig. yi. Longitudinal section of same embryo as Fig. 70. (Zeiss Dx4). 

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, Dx 4). 

cnt, entoderm-anlage ; m.v, malpighian vessels ; pt, proctodium ; other letters as in 
preceding figures. 
F'g- 73- Sagittal section through posterior portion of embryo represented in Fig. 51. (from embryo 
taken on April ist). (Zeiss, Dx4). 

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 9th. (Zeiss D x 4). 

Lettering as in preceding figures. 
Fig. 77—82. Series of transverse section through head segment of embryo as represented in Fig. 
52, showing various ectodermal invaginations. (Zeiss, Dx4). 

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 ; or, 
supraocsophagcal ganglion ; ent, entoderm ; f.n.c, frontal nerve chord ; fl. md, flexsor 
mandibulae; s.b, suboesophageal body ; s]). ms, si)lanclinic mesoderm ; sl.g, silk gland ; 
md, mandible; mx 't~'), first and second maxilla; n, new gland or hypostigmatic 
gland ; st, stomodium. 

Explanation of Plate XI. 

F'gs- 83—91. Consecutive scries 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). 

F'gs- 83 and 84, sections through anterior portion of mandiljular region ; Fig. 85 througli 
posterior portion of same region ; Figs. 86 and 87, through portion between mandible 
and first maxilla ; F'igs. 88 and 89, through second maxilla ; Fig. 90, through posterior 



Contributions to tlie Stiiilv oi Silk->Vonns. 



n; 



portion of hcail ; Fig. 91, through iirst thoracic segment, br, supraoesophageal ganglion; 
ex. md, attachment of extensor mandibulae ; fl. md, attachment of flexor mandibuiae ; 
g.f, ganglion fi'ontale ; n, new gland or hypostigmatic gland ; s.g, salivary gland : st, 
stomodium ; sl.g, silk gland; tent(i = '), first and second tentorium; other letters as 
in preceding figures. 
Kig~. 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 9th. 
(Zeiss, Dx4). 

Lettering as in preceding figures. 
Fig. 95. New gland at base of first stigma, (taken from larva of third stage). (Zeiss, Ax 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^^"-'. Portion of the new gland, highly magnified. (Zeiss Dx2). 

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 ii 

a. Vitellophags 

b. 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 th 
gland and a new gland 

B. General considerations. 

The mesoderm 

The entoderm 

Blood-cells 

Works referred to 

Explanation of plates 

Contents 



the yolk. 



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PAGE. 

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Ueber das wirksame Princip des Tuberculinum Kochii. 



VON 



NT. 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 Reintubercuh'n 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 enthiilt. Aus Tuberkelbacillenculluren hat W. Kuhne 
folgende verschiedene Proteinfractionen erhalten : Albuminat, Acroalbu- 
mose und Deuteroalbumone ; Essigsiiurefallung und Ammonsulfatfallung/ 
von denen einige beini Tiervcrsuche eine noch energischere Wirkuug als 
das ,, Reintuberculin" Koch's zeigten. Trotzdem ist er der Meinung ,,dass 
keine dieser Substanzen mehr sei, als der Triiger des Tuberculinum 
verum ; sie sind dazu unter sich in chemischen Verhalten zu verschieden 
und uns aus dem Niihrboden und als Bcstandtheile des Handelspeptons nur 
allzu bekannt." 

Auch hat Helmann eine eingehende chemische und pharmacologische 
Untersuchung iiber das Tuberculin ausgefiihrt. Er fand als dessen 
Bestandtheile : i) Albumose und zwar Protoalbumose und Deutero- 
albumose, 2) mehrere Alkaloide, 3) Extractivstoflfe, 4) Mucin, anorganische 
Saize, Glycerin und Farbstoffe. Ferncr stellte er mit dem Alcohol- 
niederschlag des Tuberculins (A) und mit dem AlcohoIfiltrat(C) Tiervcrsuche 



* I/it/tcre beidc Fractioncn w uixlcn aus all)umosofroit^- roptoiicultui" erhalten. 



I20 ^' ^itta: 

an und faiid, dass ersteres, welches den grossten Teil der Albumose 
enthalt, starke, entziindliche Local-Reaction und kein oder nur gerlnges 
Fieber, dagegen letzteres, welches besonders die Saize und nur wenig 
Albumosen enthalt, keine locale Entziindung, aber hohes Fieber ver- 
ursaclit. Helman hat liber das von ihm aus Kartofielkulturen, welche 
unter oder ohne Mitwirkung von Serum und Glycerin gewachsen waren, 
erhaltene Tuberculin, sowie iiber die Koch'sche Lymphe Studien veroffent- 
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 gewohnlichen Albumosen, also von 
Korpern, welche ohne jede specifische bacterielle Thatigkeit aus Verdau- 
ungsgemischen isolirt werden, bei an Lupus erkrankten Menschen 
deutliche locale Reaction und hiilt die Tuberculinwirkuug wenigstens zum 
Theil fiir eine Wirkung einer gewohnlichen Albumose : daher empfiehlt er 
statt des Tuberculins, welches nacli ihm ein theures, schwer haltbares 
Praeparat und noch dazu kein einheitlicher Korper ist, die gewohnliche 
Deuteroalbumose zu benutzen, welche allerdings in etwas grosseren Dosen 
(0,05-0,075 g) angewendet werden muss. Die Deuteroalbumose ist ferner 
ein voUig reines Material, welches sich leicht vollkommen salzfrei darstell- 
en liisst ; sie crhiilt sich als weisses trockenes Pulver jahrelang unverlindert 
und crlaubt cine absolut genaue Dosirung. 

Petri und Maassen zeigten, dass gewohnliciie 10 proc. Pepton- 
bouillon, welche fiir gesunde Meerschweinchen in Mengen von 4 ccm. 
eingcspritzt ohne Naclitheil war, tuberculose Tiere totetc. Bei der Section 
fanden sich in der Umgebung der tnberculosen Herde deutliche Erschei- 
nungen einer Reaction. Nach Ansicht der Verfif. ist diese Giftwirkuiig 
wesentlich dem hohen Pcptongehalt der Nahrbouillon zuzuschreiben ; sie 
liiugnen dcslialb die Specilitaet der Tuberculinwirkung, womit audi 
mchrcrc andcrc Autorcn iibereinstimmcn. 

Von Stoffen, die dem Tubcrcuh'n ahnlich wirken, werden genaniit : 



I 



Uel)er das ivirksaiiic Priucip des Tuberculimim Koeliii. 121 

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 T3'rosin (Dixon 
und Zuill). Ausserdem noch eine grosse Anzahl der verschiedenartigsten 
Stoffe vvie Thiophen, Benzol, Sulfoharnstoff, SulfoathylharnstofF, Aceton, 
Propylamin, Trimethylamin, Allylamin, Taurin, Cadaverin (Spiegler), 
kantharidinsaure Salze (Liebreich) u. s. w. 

Nach der Untersuchung Viqucrat's verhiilt sich Tuberculin, welches 
auf 150-200° c. erhitzt wurde, gegen tuberculose Tiere wie nicht erhitztes ; 
seine weitere Behauptung, dass bernsteinsaure Salze ebenso wirken wie 
Tuberculin, wurde von Hutyra sowie von mir selbst nicht bestatigt 
gefunden. 

Ruppel untersuchte Filtrate von ^Nlassenculturen des Tuberkelbacillus 
auf ihre chemischen Bestandtheile. Die Filtrate enthielten an fiiUbaren 
Substanzen vorwiegend Deuteroalbumosen, neben primiiren Albumosen 
und Plemialbumosen ; 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 Hess 
sich nur feststellen, dass den Tuberkelbacillen ein tryptisches Verdau- 
ungsvermogen zukommt, indem sie in alkalischer Losung Eiweisskorper 
bis zur Bildung von Pepton unter gleichzeitigem Auftreten von Trj'ptophan 
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- 
klilrung der Frage zu llefern. 

Elgene Untersuchungen. 

I. Herstellving des Tuberculins. 

Das bei meincn v^crglcicheiidcn \'crsuchen angewandte Koch'sche 
Rohtuberculin wurde von mir in meincn Laboratorium aus Menschen- 



122 >'. Nitta: 

tuberkelbacillenkulturen hergestellt. Die Niihrlosung bestand aus einer 
normalen Peptonrindfleischbouillon, die eine 1.6-3.^% Normalnatronlauge 
entsprechende Aciditiit besitzt und die ich mit S% 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 ^- 1 Stunde lang sterilisirt und durch sterilisirtes 
Filtrirpapier filtrirt wurde : das Filtrat wurde auf dem Wasserbade bei 
95-100^ C. in sterilisirter Porzellanschale bis auf den zehnten Theil ihres 
urspriinglichen Gewichts eingedampft. Die in dieser Weise gewonnene 
gelbbraune, syrupartige Fliissigkeit 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 44.6375^. Stickstoff 2.4750^. Asclie 6.3322%. Chlor 
2.8609%. Das Glycerin wurde nicht bestimmt. Tuberculos gemachte 
Meerschweinchen zeigten nach Injection von 0,001 ccm. starkes Reactions- 
fieber (1-2° und dariiber) : bei hochgradig tuberculosen 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 Reintuberctilin Koch's. 

Kin Teil dcs fliissigen Rohtuberculins wird mit nur i^ Volumenteilen 
absoluten Alcohols vermcngt und 24-48 Stunden stehen gelassen : es 
bildet sich cin flockiger Bodensatz ; die uberstehende Fliissigkeit wird 
abgcgossen, nun aber ein gleichcs Volum nur 60% ige Alcohol zugesetzt, 
wieder absitzen gelassen und dies 3-4 Mai wiederholt, bis der liber dem 
Nicderschlag stehende Alcohol fast farblos erscheint: nach mehrmaligem 
Auswaschen mit absolutem Alcohol wird der Nicderschlag bei 50-60° C. 
getrocknet, wobei eine sclineewcisse leicht zorrcibliche Masse resultirt. 
Dies ist das sogenannte Koch'schc Reintubcrculin (Tubcrculinum depuratum 
Koch). Tuberculose Meerschweinchen reagiren auf Injection von 0,00005 g. 
dieses Traeparats. 



Ueber das wirksiaiue Priiicii) <les Tiiberciiliniim Koehii. 123 



III. Der in absolutem Alcohol unlosliche Teil 
des Rohtuberculins. 

Anfangs inodificirte ich die Koch'sche Methode auf die Weise, dass 
ich, statt sechzigprocentigem, absoluten Alcohol anvvandte. Wird das 
Rohtuberculin mit dem mehrfachen Volumen absoluten Alkohols unter 
Umriihren vermischt, so bildet sich eiii Niedeischlag ; nach 15-24 stiindi- 
gem Stehenlassen wird die Fliissigkeit abgegossen und der zuriiclvbleibende 
Niedersclilag iiochmals init 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 50-60° C. getrocknet : es resultirt eine gclblich 
weisse leicht zerreibliche IMasse. Eine Einspritzung von 0,0005 S- Jieser 
Substanz bei tuberculosen Meerschweinchen ruft eine deutliche Steigerung 
der Korpertemperatur hervor. Es erwies sich schwiicher als das Koch'sche 
Praeparat, wesshalb ich nun Ammonsulfat anwandte. 



IV. Ammonsulfatfallung' des Tuberculins. 

Mcin Tuberculinpraeparat wird mittelst Ammonsulfatfiillung gewonnen. 
Siittigt man flussiges Rohtuberculin mit Ammonsulfat, so scheidet sich 
eine gelbbriiunliche ziihe JNIasse ab, die nach 24 Stunden gesammelt und 
einmal mit gesiittigter Ammonsulfatlosung ausgewaschen, dann zwischen 
Filtrirpapier gepresst, uud in etwas destillirten chloroformhaltigen Wasser 
gelost, hierauf so lange gegen stromendcs Wasser dialysirt wird, bis die 
Losung keine Reaction auf Sciiwefelsiiure mehr zeigt. Die Fliissigkeit 
wird dann filtrirt, das Filtrat bei 50-60° C. eingedampft und schliesslich 
mit cincm Ueberschuss von absolutem Alcohol unter Umriihren versetzt. 
Nacli 24-48 Stunden wird der voluminose gelblicli weisse Niederschlag 
abfiltrirt, 2-3 Mai mit absolutem Alkohol ausgewaschen, zwischen Filtrir- 
papier gepresst und bei 50-69° C. getrocknet. Das dabei gewonnenc 
grauweisse Pulver ist leicht loslich und enthiilt nur 1.54469^ Asche. wahrend 
Koch'sche Praeparate an 20^^ Asche cnthaltcn. Die wiisserige Losung 



124 



>. Xitta: 



dieser Substauz reagirt neutral und ist von brilunlicher Farbe. Dieselbe 
coagulirt nicht beim Kochen und wird durch Salpetersilure sowie durch 
Ferrocyankalium und Essigsaure gefallt ; der dabei gebildete Niederschlag 
lost sich beim Ervviirmen und scheidet sich beim Abkiililen wieder ab, 
Dieselbe wird auch durch Phosphorvrolframsaure, Pikrinsilure, Gerbsilure, 
Sublimat und Ammonsulfat gefallt : Biuret, Millon'sche uud Xanthoprotein- 
reactioncn fallen positiv aus. 

Dass mein Praeparat frei von peptonartigen Substanzen ist, geht 
daraus hevyor, 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 wiisserige Losung meiner Tuberculinalbumose gibt bei Siittigung 
mit Chlornatrium sehr schwachen, beim weiteren Zusatz von etwas 
Essigsiiure aber starken Niederschlag, was andeutet, dass meine Tuber- 
culinalbumose hauptsachlich den Character einer Deuteroalbumose von 
Kiihne trligt ; sie enthillt aber noch Spuren von Protoalbumose. Die 
Thatsache aber, dass eine wiisserige Losung meines Praeparates mit 
verdiinnter Essigsiiure einen schwachen im Ueberschuss derselben loslichen 
Niederschlag erzeugt, deutet darauf hin, dass sie eine geringe IMenge einer 
Albumose enthiilt, welche der Atmidalbumose Neumeister's analog ist. 



V. Injectionsversuche. 

a) Mit Meerscliweinchen. 

Die in dcstillirtem Wasser geloste Tuberculinalbumose wurde in 
verschiedenen Dosen tubcrculoscn Meerscliweinchen (Korpergewicht : ca 
400-5CO g.) subcutan eingespritzt. Die dabci gewonncnen Ergcbnissc sind 
aus folgender Tabclle 7.u erschen : 



I 



Ueber das wirksame Priucip des Tuberculin uiii Kochii. 



125 



6 
3 


1 1 






Korpertemperatui 


. Cels.= 








'A 


52 

> 


be 

c 

3 

> CU 


Nach Einspritzung (Stundeii). 




■^3 




















t-, 


U 3 


IS. 


ih 


2h 


3l^ 


4h 


5h 


6h 


7!> 


8I1 


- 


I 


0,005 


3S,5 


— 


38,8 


40,0 


40,5 


40,4 


40,1 


39,2 


38,6 


2.8 


2 


0,001 


38,7 


— 


38,4 


38,8 


39,25 


40,0 


40,0 


39,8 


39,6 


1-3 


3 


0,0005 


37,25 


37.35 


37,95 


39,25 


39,75 


40,05 


40,05 


39,85 


— 


2.8 


4 


o,ocoi 


38,2 


38,2 


38,6 


38,8 


39,15 


39,8 


40,2 


39,7 


— 


2.0 


5 


0,00005 


38,5 


38,9 


38,9 


39,1 


39,7 


39,5 


39,4 


39,3 


39,0 


1.2 


6 


0,00001 


38,6 


39,4 


40,2 


40,2 


39-9 


39.2 


39,0 


39,1 


38,9 


1.6 


7 


0,000005 


39>i 


— 


39,1 


39,3 


39,5 


40,1 


40,1 


40,0 


39,6 


I.O 


8 


0,000001 


39>o 


— 


39,0 


39,2 


39,4 


39-1 


39,1 


39,4 


39,2 


0.4 



Eei Betrachtung der obigen Versuche erkennt man, class die Tuber- 
cuHnalbumose in Dosen von 0,00001 ( — 0,000005) g. bei tuberculosen 
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 : 

Rohtuberculin nach Koch 0,001 ccm. 

Reintubercuh'n nach Koch 0,00005 g. 

Tuberculinalbumose 0,00001 g. 

Die Versuche beziigllch ihrer totlichen Wirkung auf tuberculose Meer- 
schweinchen (Korpergewicht : ca. 400-500 g.) hattcn folgendes Ergeb- 
niss : 



126 



N. Mtia: 



o 
3 




II 

^ 




Korpertemperatu)- 


. Cels.° 






2 


ho 
3 

H 

c 
W 


Nach Einspritzung. (Stunden.) 


Bemerkungen. 


y, 


2h 


31^ 


4h 


5h 


6h 


7h 




I 


34 


0,1 


38,9 


40,0 


39,6 


39,3 


— 


Todt. 




Todt. 


2 


„ 


0,05 


38,0 


38,1 


37,9 


37,5 


— 


— 


Todt. 


„ 


3 


48 


0,01 


38,3 


38,6 


39,0 


40,2 


40,0 


39,7 


37,3 


Todt in der 
folgenden Nacht. 


4 


37 


0,005 


38,6 


38,8 


39,5 


38,8 


37,0 


— 


Talt. 


Todt. 


5 


28 


0,001 


37,4 


38,2 


39,0 


39,5 


39,6 


39,4 


39,3 


Todt in der 
folgenden Nacht. 


6 


37 


0,0005 


38,5 


39>4 


39,6 


39,3 


38,2 


— 


— 


" 


7 


,> 


0,0005 


38,4 


38,6 


39,3 


40,1 


40,3 


40,6 


39,9 


Lebend. 


8 


.. 


0,0001 


37,5 


37,7 


38,7 


39,5 


40,5 


39,1 


38,7 


» 


9 


" 


0,0001 


38,2 


39,5 


39,6 


40,2 


39,8 


38,6 


39,3 


" 



Aus obigen Tabellen ersieht man, dass die minimale Letlialdose der 
Tuberculinalbumose fiir mittelgrosse Meerschweincben 28-48 Tage nach 
der Impfung init Tuberkelbacillen i Milligr. ist. Bei den Tieren, welche 
dieser Dose eriagen, finden sich bei den Eingeweiden, insbesondere an der 
Oberfliiche der Milz und Leber, zahlreiche haemorrhagische Flecke von 
Mohnsamen- bis Pfenniggrosse. Diese Haemorrhagie vvurde auch von mir 
bei der Rohtuberculininjection beobachtet : nach R. Koch ist dieser 
Befund ein charakteristisches Merkmal der Tuberciih'nvvirkung. 

Nach Koch erfordert die Totung der Tiere von seinem Reintuberculin 
5-10 Milligr: somit ist Gifcwirkung meiner Tuberculinalbumose 5-10 fach 
so stark als die der Koch'schen. 

Zum Vergleich babe ich auch einige Tierversuche mit den von mir 
aus Witte'schcm Pepton hergestellten Albumosen ausgefiihrt. Die Injec- 
tion von 0.1 g. dieses Praeparats hatte nur einige Zehntel Grade Tem- 
peraturstcigerung im Gefolge und schadigte das Wohlbefinden der Tiere 
nicht im Geringstcn, wiihrend von meiner Tuberculinalbumose schon 
0,001 g. totlich auf dicsc Tiere wirkt und 0,00001 g. schon eine Tem- 
peratursteigerung herbcifiihrt. 



I 



Uel)e» (las wirksame Priiicip des Tuberculinam Koehii. 



127 



b) Mit Rindern. 

Es dienten hier Kinder, welche ein Jahr vorher bei der Probeinjection 
des gewohnlichen Tuberculins eine deutliche Fieberreaction gezeigt batten, 
zu den Versuchen, deren Resultate in folgender Tabelle zusammengestellt 
sind : 



CO 

O 

pq 

< 
t— I 

u 
Pi 

M 

pq 



Pi; 
w 
> 






U 



a 3 



> 



■3 ■asouinqitiijino 
-.loqnjL .lop oSu3j\[ 



o 00 
o 00 






1^ " 

6 o 



Q r " 



« o 



« « o 



ro 00 



O M 

i-T o" 



►. C^ C<5 



O >-o 



~ «- o 



■.I3uuun\[ opiiojnirj 












128 



N. Nitta; 



u 

H 
O 



w 
u 

ID 

CO 

W 
> 



.0 °'''^p 




IT) 


N 


10 


CO 


fl^ 


t--. 


1J1 


co__ 


q\ 




rf 




Cl^ 


10 


JL 


unja^opjs 


i_r 


^^ 


t^ 


^^ 












c-r 


cT 


M* 


cf 


c^" 


-jn^T^jsduiaj^ 




































N 





„ 


CO 


C> 





M 


lY-1 


q. 


CO 


li-i 


CO 











'n 


0; 


cR 


c^ 


0' 


c> 


0* 


6 


CTn 


c5 


c> 


0* 


c> 


0* 


0" 






N 


rt- 


CO 


CO 


ri- 


CO 


rj- 


rj- 


CO 


CO 


CO 


rf 


CO 


rf 


rf 








ro 


rt- 


rt- 




N 




CO 


00 


t^ 


t^ 


t^ 


CN 


NC 


VO 






"o 








0" 


^ 


0" 


0" 


0" 


cR 


6 


0* 


0* 


d 


6 


6 






M 


'^ 


rf 


^ 




rf 


n- 


rf 


CO 


rf 


rf 


rf 


rf 


rf 


rf 
































U-) 


U-) 






r* 


fO 


CO 


to 


CO 


u-l 


CO 


i-^ 


rf 


r-^ 


t^ 


CN 


t-H 


CO 


CN 




^^ 


CC 


0" 


0" 


0" 


^ 


0" 


o" 


o~ 


0* 


0* 


0* 


c* 


H-T 


0* 


0" 




^ 




■* 


rf 


rh 


n- 


ri- 


rf 


rf 


rf 


rf 


rf 


rf 


rf 


rf 


rf 




5 






























































u-i 


10 




x 


^ 


\o 


00 


r^ 


c> 


00 


oo_ 


rf 


vD 





r- 


CN 


CO 


iJ-i 














































0^ 


0" 





0' 


0" 


c^ 


d 


0" 


6 


0* 


d 


0* 





1^ 






l-l 


CO 


rt 


CO 


rt- 


ri- 


CO 


rf 


rf 


rf 


rf 


rf 


rf 


rf 


rf 




t/. 


































c 


































3 




































































.-^ 






























u-l 




^ 


^ 


l-l 


q_ 


r-« 


Cn 


CN 


r^ 


ro 


H^ 


vo 


q. 


CN 


q. 


On 


t^ 




































^ , 


^ 


'i- 





t-^ 


CiN 


0" 





0" 


0" 


1^ 


Cj^ 


hT 





hi 


hH 


»M 


F^ 


j^ 




'4- 


•* 


CO 


rl- 


■^ 


rt- 


rf 


rf 


CO 


rf 


rf 


rf 


rf 


rf 





S 





































































































J_' 


r; 






























u-l. 


^ 


;^ 


r-- 





M 


CO_ 


f) 


q_ 





u-i 


CO 





CO 





►-( 


CO 


M 


rt 




Tj 


0" 


0" 


c> 


(_r 


i-T 


0' 


0" 


0' 


(> 


i-T 


i_r 


0* 


0* 


i-T 


i^ 




t-< 


M- 


><*• 


CO 


•<j- 


rj- 


rt- 


rf 


rf 


CO 


Tf 


rf 


rf 


rf 


rf 





































Ci, 


































p 


































_o 
































































vo 









^ 





I^ 


CO 


t^ 


N 





q\ 


)-i 


r--. 


CO 


M 





r^ 


On 











































00" 


co' 


co" 





0' 


0" 


cf^ 





00 


0* 





c5 





0* 


:o 






m 


CO 


CO 


CO 


rl- 


rf 


CO 


rf 


CO 


rf 


rf 


CO 


rf 


rf 


>■> 






























































































VO 


vo 








rt- 


vo 


Krf 


t~. 





"^ 





N 


lO 


N 








rf 


CO 








































c^ 


00" 


CO 


CO 


oc 


0" 


tC 


di 





00 


ON 





d 












CO 


CO 


CO 


CO 


rt- 


CO 


CO 


rf 


CO 


CO 


CO 


CO 


rf 


rf 






p- 


CO 


00 





r^ 


CO 








CO 


J^ 




NO 





r~. 











































'0 "^ 


00 


CO 


co' 


CO 


00" 


c> 


cf^ 


C^ 


CO 


ON 


CO* 


d 


CN 


ON 






rt 


CO 


CO 


CO 


CO 


CO 


CO 


CO 


CO 


CO 


CO 


CO 


CO 


CO 


CO 




:/; 


"A 
































^ 


































p 






































































































'^ 


tc 


00 


t^ 


co^ 








t~» 


^ 


0^ 


co^ 


c^ 


vO_ 


t-~ 


1,^ 


^ 






« N 


































CO 


JO 


00" 


d» 


00* 


00 


00' 


&• 


CO* 


c? 


00* 


00* 


CN 


CTi 




r^ 


."ti *^ 


CO 


CO 


CO 


CO 


CO 


CO 


CO 


CO 


CO 


CO 


ro 


CO 


CO 


CO 




y 


^ 
































;_ 


































I-' 




































;^ 


t^ 


CO 


00 


t^ 


■<*- 





rf 


»iN 


vo 


q 


o_ 


CO 












































CO 


00 


00" 


00 


00 


c» 


CO* 


ON 


CO* 


On 


CO* 


00 










ro 


CO 


CO 


CO 


CO 


CO 


CO 


CO 


CO 


CO 


CO 


CO 


1 


1 




•UIDD 






























■ui 


[nDJoqnj_ 


6 


r 


r 


r 


r 


s 


r 


r 


s 


5 


t 


= 


r 


5 


S3 


p 03llDJ\; 




























■jouiui 


ns^ Dpi 


.ijni:'i 


" 


n 


CO 


rf 


u-i 


vO 


t^ 


00 


ON 





- 


M 


CO 


•* 



Ueber das wirksame Princii) des TubereuHnnm Kochii. 120 

Bei diesen Fallen wurde keine Section ausgefiihrt ; dcch ist diese 
Nachpriifung hier entbehrlich, denn der vorliegende Versuch zielt nicht 
auf die Feststellung der Diagnose der Tuberculose ab, sondern er be- 
zweckt, die Analogte der Wirkung zwischen beiden Praeparaten zu 
constatiren. Man erkennt, dass sainmtliche Tiere, welche auf Rohtuber- 
culinimpfung reagirt liaben, eine starke Fieberreaction auf Einspritzung 
meiner Tuberculinalbumose zeigen. 

Im Anschluss an diese Tierversuche habe ich nocli mit anderen 
aus dem Rohtuberculin hergestellten Korpern und zwar mit dem Aether- 
extract, welches aus dem mit verdiinnter Schwefelsiiure versetzten Rohtu- 
berculin gewonnen wurde, sowie mit dem direkt hergestellten Aether- 
extract an tuberculosen Meerschweinchen (mittelgross) Versuche gemacht: 
dabei liess sich aber nirgends eine tnberculinahnliche Wirkung beobachten. 
Ferner zeigten tuberculose Meerschweinchen (mittelgross) nach Einspritz- 
ung von Bernsteinsiiure (0,01-0,05 g.) oder von bernsteinsaurem Natron 
(0,01-0,1 g.) gar keine Steigerung der Korpertemperatur, was die Behaup- 
tung Viquerat's, dass diese Saure das active Princip im Rohtuberculin sei, 
widerlesrt. 



VI. Chemisches Verhalten ineiner Tuberculinalbumose. 

Eine 2% ige Losung der Tuberculinalbum.ose zeigt folgendes Ver- 
halten : 

1. Alcohol 96^^ : Mit gleichem Volum fallen zarte, weisse Flocken aus. 

2. Salpetersiiure in der Kiilte : Niederschlag, der sich in der Wiirme 
rasch lost, in der Kiilte aber wieder auftritt. 

3. Gleiches Volumen conzentr. Kochsalzlosung zu der mit Essigsiiure 
angesiiuerten Losung gesetzt : miissiger Niederschlag, beim Er- 
hitzen nicht loslich. 

4. Sattigung der neutralen Losung mit Kochsalz : Spur Triibung. 

5. Verdiuinte Kupfersulfatlosung : starke Fallung. 

6. Essigsaure-FerrocN'ankalium : starke Fiillung, loslich beim Erw Lir- 
nien. 



I 



130 



7 



N. Tiita: 



8 



lO 

II 

12 

13 

M 
15 

i6 



Pikrinsiiure : starke Fiillung. 



Metaphosphorsaure : Fiillung, im Ueberschuss wieder loslich. 
Trichloressigsaure : starke Fiillung, in der Hitze loslich, in der 
Kiilte wiederkehrend. 

Jodquecksilberkalium : erst in saurer Losung starke Fiillung, 
welche sich im Ueberschuss von Salzsiiure nicht lost, 
Gerbsiiure : starke Fiillung, die sich in der Hitze nicht lost. 
Millon'sches Reagens : weisse Fallung, beim Kochen Rothfiirbung. 
Xanthoproteinprobe : starke Fiillung, welche sich im Ueberschuss 
von Salpetersiiure lost, beim Erhitzen Gelbfiirbung. 
Adamkiewiez'sche Reaction : Violettfarbung. 

^lolisch'sche Zuckerprobe : schwach positiv, beim Erhitzen 
schwache Violettfiirbung. 
Biuretprobe mit Kupfersulfat : positiv. 

Biuretprobe mit Nickelsulfat : beim Erwiirmen schwache Gelb- 
fiirbung. 
i8. Kochen mit Alkali- und Bleiacetat : Schwarzfiirbung. 

Nach diesen Reactionen entspricht meine Tuberculinalbumose am 
nachsten der Deuteroalbumose Kiihne's, ferner in einem gewissen Grade 
der secundiiren Albumose A von Pick ; jedoch mit dem Unterschied, dass 
meine Albumose bei den oben unter 3 und 6 erwiihnten Reactionen einen 
starken Niederschlag gibt, wiihrend die secundiire Albumose A Pick's 
hierbei nur Spur Triibung gibt ; die secundiire Albumose B. und C. gibt 
hier gar keine Triibung. 



VII. Hitzebestandigkeit der Tuberculinalbumose. 

Eine i% ige wiisserige Ecisung der Tuberculinalbumose wurde in 
kochcndem Wasser je 10 Minuten, 30 Minuten und eine Stunde lang 
erhitzt, dann tuberculosen Meerschweinchen subcutan eingespritzt ; fol- 
gende Tabelle zeigt die dabei gewonnenen Ergebnisse : 



I 



Ueber das wirksaino I'riiioip Ues Tuberciiliuum Kocliii. 



131 



. 


Erhitzuiigs- 
dauer. 


^1 


Korpcrtemperatur. Cels° 




-1 


\'( ir 


Nach Einspritzung, (Stunden.) 






fa i; 




S 3 


Einspritz- 
ung. 








t tf. 


^-^ 


ih 


2\\ 


31^ 


4h 


5h 


6h 


7h 


r- 


I 


Zelin Minuten. 


0,001 


38,3 


— 


39.8 


40,0 


39,9 


38.8 


38,3 


— 


1-7 


2 


.. 


0,0005 


38,0 


— 


39.2 


40,0 


40,3 


39.9 


38,8 




2.3 


3 


„ 


0,0005 


37,7 


— 


38,4 


40,2 


40,5 


40,3 


39,7 


38,6 


2,8 


4 


Dreissige 
Minuten, 


0,0005 


37,9 


— 


39,6 


40,35 


40,15 


40,15 


40,3 


40,0 


2.45 


5 


» 


0,00035 


38,4 


— 


39,4 


38,8 


39,5 


40,15 


40.0 


39,8 


1,75 


6 


Ein Stunde. 


0,0005 


37,-5 


— 


39-8 


40,4 


40,25 


39,0 


38,8 


— 


3.0 


7 


„ 


0,00025 


37,6 


— 


39.6 


40,2 


40,1 


40,0 


40,2 


40,0 


2,6 


S 


Controllosung. 


0,0005 


37,7 


— 


38,1 


40,3 


40,2 


40,0 


39,6 


38,8 


2,6 


9 


'• 


0,0005 


37,4 


— 


38,9 


40,45 


40,7 


40,25 


39.5 


38,25 


5.3 



Es unterliegt also kaum einem Zweifel, dass die Tuberculinalbumose 
gegen Hitze selir widerstandsfiihig ist. Eine i stlindige Erhitzung bei 
lOcP C. ist nicht im Stande, die Wirksamkeit der Tubercidinalbumose im 
Geringsten zu schiidigen. 



VIII. Verhalten der Tuberculinalbumose gegen 
Pepsin und Trypsin. 

Je 0,1 g. Tuberculinalbumose wurde einerseits in 10 ccm. o,2^y iger 
Salzsiiure und andererseits im gleichen Volum o,2J?o iger Sodalosung 
(NagCOg) gelost und dort etwas Pepsin, hier aber ebensoviel Trypsin 
zugesetzt. Bei Gegenwart von etwas Chloroform wurden beide Losungen 
wohlverschlossen 2 Tage lang im Brutofen bei 37-38° gelassen. Ilierauf 
wurden sie im Dampftopf 10 Minuten lang erhitzt und tuberculosen Meer- 
sclnveinchen subcutan cingespritzt ; gleichzeitig wurden Controllosungen 
(Tuberculinalbumoselosungen in 0,2^^ iger Salzsiiure und in o,2^q iger 
Sodalosung ohne Pepsin resp. Trypsin) gepriift : Die Ergebnisse sind in 
folgendcn Tabellen ,-^usammengcstellt : 



132 



X. >'itta: 





Tuberculm- 
albumose. 


3 S 

W IS 

S3 


Korpertemper 


itur. Cels.° 






u 

V 


-1 


Vor 

Einspritz- 

ung. 


Xach Einsp 


ritzung. (Stunden.) 






ih 


21: 


3li 


4h 


.11 


6I1 


7h 


S 3 


I 


l\Iit Pepsin. 


o,ooi 


38,0 


— 


38,8 


38,9 


38,7 


38,5 


38,3 


38,3 


0,9 


2 


)» 


0,0005 


37=4 


— 


37,7 


38,25 


38,4 


37,75 


38,0 


37,9 


1,0 


3 


» 


0,0001 


37,7 


— 


38,0 


38,45 


38,6 


38,6 


38,3 


38,0 


0,9 


4 


Ohne Pepsin. 


0,001 


3S,i 


— 


38,2 


40,0 


39,9 


39>5 


39,3 


39,3 


h9 


5 


» 


0,0002 


37,7 


— 


38,0 


40,0 


40,3 


40,1 


39,7 


39,5 


2,6 


6 


j\Iit Trypsin. 


0,001 


38,0 


— 


37,9 


38,3 


38,6 


38,5 


38,0 


38,0 


0,6 


7 


.. 


0,0005 


38,3 


— 


38,2 


38,5 


38,7 


38,8 


39,1 


38,9 


0,8 


8 


» 


0,0001 


38,3 


— 


38,8 


39,1 


39,0 


39,1 


38,8 


38.25 


0,8 


9 


Ohne Trypsin. 


0,001 


37,4 


— 


38,2 


39>o 


39,5 


39.6 


39,4 


39,3 


2,2 


lo 


» 


0,0002 


37,9 


— 


37,9 


38,3 


39,6 


39,4 


39,5 


39,1 


1,7 



Die Tiere verhalten sich also bei Behandlung mit verdauter Tuber- 
culinalbumose indifferent ; Pepsin und Trypsin batten die Tubercuh'n- 
albumose verandert. 



IX. Sind gewisse labile Atomgruppen die Ursaehe 
der Gif twirkung ? 

Die schon erwahnte Thatsachc, dass die TubercuHnalbumose bei 
einstiindigem Erwiirmen nicht im Geringsten an Wirksamkeit einbiisst, 
liess es von vornehercin wenig wahrscheinh'ch erscheinen, dass die Wirk- 
samkeit auf besonders labilen Atomgruppen beruhc. Nichtsdestoweniger 
wurden einige Versuche angestellt mit Korpern, welche sehr leicht in 
labile Amido- und Aldehyd- oder Keton- gruppen eingreifen. 

Je 5 ccm. 20^5^ ige wiisserige Losung des Rohtuberculins wurde mit 
25 ccm. der folgenden Losungen vermischt (in jcdem Falle zwei Kolbchen). 

1. Controllosung. 

2. !_%' Natriumnitritlosung : unmittelbar vor Gebrauch mit einigen 
Tropfen verdiinntcr Essigsiiure versctzt. 



i 



Uel)er das wirkstaiue Priiicii) des Tiil)ci'culiniiui Kocliii. 



133 



3. s% Formaldehydlosung-. 

4. 1% Hydroxylaminchloridlosung' : unmittelbar vor Gebrauch mit 
Natriumcarbonat neutralisirt, (eiitspr. 0,47 freiem Hydroxy lamin). 

Die Mischungen wurden mit etwas Chloroform versetzt und 24 Stunden 
bei Zimmertemperatur stehen gelassen, dann mit absolutem Alcohol 
ausgefiillt, der Niederschlag wiederholt mit absolutem Alcohol ausge- 
waschen und uach Pressen zwischen Filtrirpapier in wasserigen Losungen 
tuberculosen Meerschweinchen subcutan eingespritzt. Die dabei gemach- 
ten Beobaclitungen zeigt die folgende Tabelle : 





Reagenticn. 


3 

en 

n3 « 


Korpeitemperatur. Cels° 






3 

OT 




Vor 


Nacli Einspritzung. 


(Stunden.) 




in 0) 






Einspritz- 
ung. 














ti 


ih 


2h 


3'^ 


4h 


51^ 


6h 


7h 




I 


Controllosung. 


0,04 


38,6 


— 


39,0 


40,8 


40,6 


40,4 


40,0 


37,0 


2,2 


2 


Natriumnitrit. 


0,2 


38,4 


— 


38,8 


39,2 


39.9 


39,7 


39,7 


39,6 


1,5 


3 


" 


0,1 


37,8 


— 


38,65 


38,8 


40,0 


39,7 


40,0 


39,6 


2,3 


4 


" 


0,04 


3S0 


— 


39,0 


39,3 


39,7 


39,4 


39,0 


39,0 


1,2 


5 


Formaklehyd. 


0,2 


37,9 


— 


37,9 


40,0 


40,3 


40,0 


40,0 


40,0 


2,5 


6 


» 


0,1 


38,2 


— 


38,1 


39,4 


39,6 


39.5 


39,2 


39,2 


1,4 


7 


» 


0,04 


38,5 


— 


38,2 


39,4 


40,2 


40,2 


39,8 


39,5 


1,7 


8 


Hydroxyl- 
amizi chloric!. 


0,2 


37,9 


— 


38,0 


38,3 


39,5 


39,7 


39,7 


40,2 


2,3 


9 


» 


0,1 


38,0 


— 


38,15 


39,2 


40,0 


40,5 


40,3 


40,2 


2,5 


10 


» 


0,04 


38,6 


— 


39,0 


39-7 


40,1 


40,4 


40.1 


39,7 


i,S 



Hieraus ersieht man, dass oben erwahnte Reagentien keinerlei schiidi- 
genden Einfluss auf das Tuberculin hatten, dessen Natur demnach 
wohl verschieden ist von derjenigen der gewohnlichen Enzyme ; denn 
diese werden durch S% Eormaldehyd nacli 24 Stunden stehen leicht 
unwirksam. 



134 >. Nitta 



Schlussfolgerungen. 

Die Resultate meiner Untersuchungen sind wie folgt : 

1. Die Ammonsulfatfallung des Tuberculins iiussert bei Meerschwein- 
cben und Rindern sich qualitativ wie das Koch'sche ReintubercuHn ; 
es reichen jedcch von meinen Praeparat weit geringere Rlengen (|) 
bin, dieselbe Reaction zu erzeugen. 

2. Die Ammonsulfatfallung bestebt wesentlicb aus einer Deuteroalbu- 
mose nebst Spuren Prot- und Amidalbumose (Tubercub'nalbumose 
Nitta's). 

3. Unter der Einwirkung von Pepsin und Trypsin verliert meine 
Tuberculinalbumose ihre specifische Wirkung. 

4. Nacb einstiindigem Erbitzen auf 100° C. bebiilt die wiisserige Losung 
der Tuberculinalbumose ihre specifische Wirkung. 

5. Die specifische Wirkung des Tuberculins veriindert sich nicht im 
Geringsten unter der Einwirkung von Natriumnitrit (1%), Formal- 
dehyd (5%) und Hydroxylamin (0,47^). 

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 
,, Tuberculins," ist ganz specifisch, und durch gewohnliche Albu- 
mosen nicht herbeizufuhren. 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 gutige Unterstutzung 
meinen ergebensten Dank auszusprechen. 



I 



Ueber das wiiksfline rriiicip des TiiHerculiuuiii Koeliii. 135 



Lileralur. 

1. Buchner, H., Tuberculinrcaction cUirch ProteVne nicht specifisclur 
Bacterien, Miinch. Med. Wochenschr. 1891. No. 49. 

2. Dixon u. Zuill, Reaction of the Amido-group upon the Wasting Animal 
Economy. Philadelphia (The Amer. Med. Pre.ss Comp.) 189F. 

3. Helman, C, Des Proprietes de la tuberculine provenant de Bacilles 
tuberculeiix cultives sur pommes de terre, Arciiiv de.s Sciences 
biologiques publ. p. I'lnstitut imper. de med. e.xper. a St. Pcters- 
bourg. T. I. No, i u. 2. 

4. Hunter, W., On the Nature, Action and Tlierapeutic Value of the 
Active Principles of Tuberculin, Brit, Med, Journ. 1891 Jul}-, 25. 

5. Hutyra, F., Tuberculinversuche bei Rindern : Nachtrag, Zeit- 
schrift f. Thiermedicin, Bd. IV. Heft, i, 

6. Klemperer, G., Die Beziehungen ver.schiedener Bakteriengifte zur 
Immunisirung und Heilung, Zeitschrift f. klin, Medicin, 15d. XX. 
Heft. I u. 2. 

7. Koch, R., Weitere Mitthcilung iiber ein Heilmittel gegen Tuber- 
kulose, Deutsche medicin. Wochenschrift, 1890, No. 46 a. 

8. Derselbe, Fortsetzung der Mittheilungen iiber cin Heilmittel gcgcn 
Tuberkulose. Ebendas. 1891. No, 3. 

9. Derselbe, Mittheilungen iiber das Tuberkulin. Kbendas. 1S91. 
No. 43. 

10. Kiihne, W., Weitere Untersuchungen iiber die Proteine des Tuber- 
culins, Zeitschrift f. Biologic, Bd. XXX. (N. F. Bd. XH.) 

11. Liebreich, O,, Ueber Lupusheilung durcl^ Cantharidin uiul iiber 
Tuberkulose, Berl. klin. Wochenschrift, 1895, N^o. 14, 15. 

12. Matthes, M., Ueber die Wirkung einiger subcutan einvcrleibten 
Albumosen auf den tubcrculos inficirten (Vganismus, Deutsche 
Archiv f. klin. Medicin, Bd. 54, Heft, i, 

13. V, Mosetig, Teucrin, Wiener med. Presse, 1893, No. 6. 

14. N^eumeister, R., Lehrbuch der physiolog. Chemie. 1897. 

15. Petri u. Maassen, Beitrlige znr Biologic der krankheitserregenden 



1 



1^6 -^« Nitta: Ueber das wirksame Priiicii) des Tiiberoiiliiiiim Kocliii. 

Bakterien, insbesondere iiber die Bildung von Schwefehvasserstofif 
durch dieselben unter vornehmlicher Beriicksichtigung des Schwei- 
nerothlaufs, Arbeiten aus dem kaiserl. Gesundheitsamte, Bd. VIII. 
i6. Pick, E., Untersuchungen iiber die Protein. stoffe, Zeitschrift. f. 
physiolog. Chemie, Bd. XXIV, Heft. 3. 

17. Roemer, G., Tuberculinreaktion durch Bacterienextrakte, Wiener 
klin. Woclienschrift. 1891, No. 45. 

18. Ruppel, G., Zur Chemie der Tuberkelbacillen ; Erste Mittlieilnng, 
Zeitschrift f. physiolog Chemie, Bd. XXVI. Heft. 2 u. 3. 

19. Spiegler, E., Ueber Localreaction in Folge liypodermatischer 
Einverleibung chcmischer Verbindungcn, Centralblatt f. kh'n. 
Medicin, 1893, No. 36. 

20. Viquerat, Beitrag y.ur Tuberculinfrage, Centralblatt f. Bakt., Bd. 
XXVI. No. 10. 



Ueber Ernahrungsverhaltnisse beim Bacillus 
prodlglosus. 

VOIV 

O. Loew und Y. Kozai. 



Da aus inehreren Beobachtungen die Bildung eines bacteriolytischen 
Knzyms beim Bac. pyodigiosus wahrsclieinlich wiirde, stellten wir einigc 
VersLichc an, wclchc Aufkliirung dariibcr geben sollten, inwicueit die 
Bildung cincs solcheii Enzyms mit Eniahrungeverhaltnissen bei diesem 
Microbcn zusammenhaiigt. Fermi bcobaclitete, dass Zucker mit Ammoniak- 
salzen die Bildung cincs protcolytischen Enzyms verhindert, dagegen 
Glycerin mit Ammoniaksalzen dieselbc bcgiinstigt. Ob dieses proteoly- 
tisclie Enzym zugleich ein bacteriolytisclies ist oder neben dem ersteren 
cin bacteriolytisclies vorhanden ist, wurdc nicht untersucht. Die Existcnz 
eines bacteriolytischen Enzyms in Culturen das B. prodigiosus schien aus 
der Beobachtung von FreudenreicJi' hervorzugehen, dass diese Culturen 
sehr entwicklungshemmend auf einige anderc Bacterienarten wirken. 

Bci unsercr crsten Versuchsrcihc verwcndeten wir folgendc 
Loesungen : 

I. Pepton \% mit Glycerin o.\%. 

3. ,, ,, mit essigsaurem Natron 0.2% und Asparagin 0.2%. 

4. Pepton 0.1% mit essigsaurem Natron \%. 

5. Asparagin 0.2% mit Glycose \%. 

6. Harnstoff 0.2% mit Glycose \%. 

7. Natriumnitrat 0.2^ mit Glycose \%. 

8. Bouillon. 

Die zugesetztcn Mineralsal/c bestanden hier aus : 

1 Arch liyg. Bd, 14 ; S. 16 und 30. 

2 Jahresber. f. Bakt. iSSD, S. 531. 



I ^8 0. Loew und Y. Kozai : 

Secundarein Kaliumpliosphat 0.2%. 

Natriumsulfat OA%. 

Magnesiumsulfat .... 0.01^. 

Die Proben wurden zuerst zwolf Tage bei 10-15 , danii iioch fiinf Tage 
im Brutkasten gehalten. Es ergab sich dann folgendes Resultat : 
Loesung i utid 2 : Viel Bacteriensedimcnt, und wenig Farbstoft". 

3 : Nach anfilnglich relclilicher und rot gefiirbler Vegetation 
fast vollige VViederloesung. 
4. 5 und 6: Geringe Kntwicklung. 
7 : Gar keine Entvvicklung. 

8: Miissige Entwicklung, rotes Sediment,^ \velches selbst nach 
aclit 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 unsrcr zweitcn Versuchsreihe ersetzten wir das schwefelsaure 
Natron durch Chlornatrium, machten ferner die Loesungen schwach 
alkalisch und niacliten geringe Zusiitze von Stoffen, welche bei hoheren 
Concentrationen giftig wirken, um zu beobachten, ob eine gunstige 
Einwirkung auf Wachsthum und Enzymbildung stattfinden wiirde.'^ 
Unsre Controlloesung hatte die folgende Zusanimensetzung : 

Pepton 0.$%. 

Glycerin o. i , , 

Dikaliumphosphat o. i ,, 

Natriumbicarbonat o.I ,, 

Natriumchlorid 0.2 ,, 

Magnesiumsulfat O.oi j^. 

1 Nach A'liiiliL- sind .Magnesia sowohl wic SchwcfdsJiure wescntlich fiir die Farbstoffproduclioii. 
Jcdcnfalis existireii aber l\ier audi nocli inaiiclic niKlcrc Eindiissc, audi solchc, weldic der rroductioii 
cntj^egen^wirkc'ii. 

2 Nadi Hi'tpf'c's bioloj^isdicni Grundgcsctz wircken Giftc lici sdir holier Verdliniiuni:; als 
Kcizmittel, 



Ueber Eriiahnin^sveiiiiiltnisse beim Bacillus itrodigioiis. i ?n 

In Loesung 2 war das Chlornatrium durch die aequivalentc Menge (0.25^) 
Natriumsulfat, In 3 durch die aequivaleiite 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 warden am 16. Januar 
inficirt und bei 6-15° C. stelicn gelassen. Am 27. Januar war der Stand 
folgender : 

1. (Control) : Keinc Haut, nur Trlibung uud ein King am Rande. 

2. (Na3S04) : Roter Ring, Spur Haut, Triibung. 

3. (NaNOy) : Loesung klar, keine Haut, nur schwaclier weisser 

Ring. 

4. (NaJ) : Triibung, schwache rote Haut. 
5- (NaF): „ 

6. (Kfcy) : Triibung, stark entwickeltc rote Haut. 
Am 3. Februar wurde bemcrkt, dass die Fiirbung am intensivsten in 5. 
war, diesc verblasste aber spilter wieder ; die Vegetation war am 
iippigsten in 6. 

Am 18. Februar war der Stand folgender : 
Bei 3. Triibung, geringer weisser Bodensatz , 

1 und 5 : Massiger Bodensatz, kaum gefiirbt. 

2 und 4 : Etwa ebenso starkes Wachstlnim als bei i und 5, aber melir 

Farbe. 
6 : Der Bodensatz betrug bier, dem Volumen nach abgeschiitzt, 

mindestens des vicrfacJie der in i. gebildeten Masse ; es war 

also eine Rei"zvirkung des FerrocyiDikaliiDiis auf die Wachs- 

thumsintensitaet unverkennbar. 
Wiihrend in der ersten Versuchsreihe das Natriumnitrat als untauglichc 
Stickstoffquelle erschien, hat es sich in dicser als sehr hemmend selbst bei 
Anwesenheit von Pepton erwiesen.' Fluornatrium und Jodkalium haben 

1 Da die Henimung von Anfaiig an vorhandcn war, so kann sic nicht ctwa die Folgc von erst 
gcbildctem Nitiit seiii. Die Ursachc ist nicht Icicht anzugcbcn, cs mag aber darauf hingewicsen 
werden, dass Nitrate audi hemmend auf die Entwicklung der Lcgumisnosenbacterien \\ irken {^Marchal, 
Compt. rend. 133 p. 1032) sowio auf die Kolilcnsaurcassimilation der Mcercsalgcn {Aihci\ Bot.in. 
Ccntralbl. 1902 p. 120) und schliesslich audi auf die katalytische \Virkuni^ der Katakase. 



I40 



0. Loew imd Y. Kozai 



in dcr aiigewandteii Verdunnung das Waclistluim nicht gefordert, wenig- 
stens nicht in direct erkennbarem Maasc. 

Die auffallende VVirkung des Ferrocyankaliums vcranlasste nns zu 
eiiiem weiteren Versucli, in welchem diese Wirkung aufden B. prodigiosus 
mit der auf anderc Microbenartcn vergHchcn wiirdc. Als Niihrloesung 
dicntc hier Bouillon, jc (S cc. in ciner Eprouvettc. 

Nach drci Tagen bei 35° war das Resultat folgendes : 





Bouillon. 




Ohne Zusatz. 


Mit o.oi_?^ Ferrocyankalium. 


B. prorligiosus. 


Keiii Bodensatz, schwaclicr rutcr 
Ring. 


Starker Botlcnsatz, dicker rotcr 
Ring. 


H. } cyoyuncus. 


Dicke llaut. 


Schwache Haut. 


B. mesenter. ruber. 


Starke Ilaut. 


Schwache Ilaut. 


B. megatherium. 


I Taut, 


Xur Triiljung und Ring. 


B, Zeiikeri. 


Triihuug u. Flockcn. 


Triibung und Flockcn. 


li. cyanogeinis. 


Haul uml Botlcnsatz. Duiikle 
]'"arl)ung nahc dcr Oljcrfinclie. 


(leringe Ilaut und Bodensatz, 
schwaclierc Farbung. 


B. capsulatus. 


Triibung, starker Rand. 


Tiiibung, schwacher Rand. 


I!. acMi lattici //////,•. 


Starker ISodensatz. 


ScUwaclier Bodensatz. 


B. subtilis. 


Dickc Ilaut. 


Diinue Haut. 


B. tyj)bi niur. 


rrul)ung. 


I'ruliung. 



I'^s hat sich also eine stinuilircndc Wirkung niu- beim 1^. prodigiosus 
erkcnncn lasscn, in den andcrn gepriiften Fallen ergab sich nieisteiis einc 
dcutlichc Schadigung. Dieses Resultat veranlasst tins zur Annahmc, dass 
beim V). prodigiosus es sich uni eine Spaltung des Ferrocyankaliums 
handclt, wobei cinerseits die schiidigcnde Cyanwasserstoffsiiure sofort 



i 



lober Enuluungsvorhiiltnisso beiiu Bacillus |>rodii^ioii< 



141 



weiter zersetzt wird, andrerseits das in den Zellen freiwerdende Eisen in 
eiiicr lockeren salzartigcii Verbindung eine fordenide Wirkunf^ ausiibt. 

Fassen wir unsre Beobachtungen am B. prodigiosiis kiir/. zusammeii. 
so ergibt sich Folgendes : 

1. Eine fiir die Production von Farbstoff und bacteriolytischem Enzyni 
giinstige Nahrstoffcombination besteht aus Pepton \%, essigsaurern 
Natron 0.2^ und Asparagin 0.2^. 

2. Eine bctriichtliche Vermehrung stickstofffreien Materials gegeniiber 
stickstoffhaltigem iibt auf die Entwicklung eincn ungiinstigen Effect 
aus. 

3. Natriumnitrat ist nicht nur unfahig, als Stickstoffquelle zu dienen. 
sender n hemmt sogar die Entwicklung bei Gegenwart von Pepton. 

4. Jodkalium und Fluornatrium in einer Verdiinnung von o.i p.m. iiben 
keine deutliche Reizwirkung aus. 

5. Ein Fcrrocyankaliumzusatz von 0.1 p.m. befordert die Entwicklung, 
was bei andern Microben niclit zutrifft. 



Ueber die Vertheiiung des Kalks im thierischen Organismus. 



VON 



M. Toyonaga. 



Die Wichtigkeit des Kalks fiir alle thierisclien Organismen ist seit 
lange anerkaiint. Uiigefahr drci Viertel siimmtlichcr MincralstofTc der 
hoheren 'riiicrc bestclit aus Tricalciumphosphat, da dieses Salz die 
Hauptmassc der Knoclicn and der Ziiline ausmacht. Aber abgesehen von 
dieseni niehr in die Aiigen fallenden Vorlcommen linden sich noch Kalk- 
verbindungen in siinimtlichen Organen vor, ja hochst walirsclieinlich 
existiert audi bei den niedersten thierischen Organismen keine Zelle, die 
frei von Kalkverbindungen ware. Auch die grosse Wichtigkeit fur 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 unloslichen Formen Calciumfluorid oder 
Kalkoxalat iibergefiihrt \vird. Auch mag darauf hingewiesen werden, 
dass in kalksalzfreicr Losung auch das Casein der Milch auf Labzusatz 
iiicht gerinnt, wohl aber wird dabei das CaseVn so verilndert, dass nun auf 
Zusatz von Kalksalzen cin Niederschlag yon den Eigenschaften des 
frischen Kiises entsteht. Cavazzani hat es ferner wahrscheinlich gemacht, 
dass Kalksalzc auch fiir die Gerinnung des Muskelplasmas von Bcdeutung 
sind. Im Harne findet sich Kalk regelmilssig gelost, ferner ausnahnisweise 
in Form von Concretionen. 

Die Anwesenheit von Kalk in den Nahrungsmitteln ist bei deni 
Bediirfniss von Thier und Mensch fiir Kalk von grosster Wichtigkeit. 
Junge Thierc leiden bei kalkarnier Nahrung an Rachititis ahnlichen 
Veriinderungen, aber auch ausgewachsene Thiere leiden in Folge eines 
solchen Mangels, da die Kalkausscheidung durch die Niere regclmassig 
fortdauert. Bunec hat deshalb auf die Gefihr hineewiesen, wenn man die 



144 ^' Tojouaga: 

an Kohlehydrat reichen Vegetabilien der meiischlicheii Nahrung durch 
blosen Robrzucker ersetzt. Erstere enthalten verschiedene tniiieralischc 
Niibrstofie wic Phosphate, Kalk, Magnesia und Eisen- Salze, wiihrend 
dem Robrzucker des Handels, der reinen Saccharose, alle diese Stoffe 
mangehi, besonders aber fiillt der Mangel von Kalk und Eisen ins Gewicht. 
Bunge glaubt, dass Aniimie und Zahncaries auf den mit Zuckergenuss ver- 
bundenen Kalk- und Eisenmangel zuriickzufiihren sind und cr schliigt 
desshalb vor, das Bediirfniss der Kinder nach Siissigkeiten, statt mit 
blosem Zuckcrpraeparaten, mit siissen Friichten, frisch oder gekocht, zu 
befriedigen. Auch fiir unsere Hausthiere ist es von sehr grosser Bedeutung, 
dass bei Zusammensetzung ciner Futterration der Kalk dieselbc Beruck- 
sicktigung ais andre Niihrbestandtheile findet. Vielfach ist Knochen- 
briichigkeit des Rindsviehs auf cine ungeniigende Ernahrung mit Kalk 
zuriickzufiihren. Die Thiere werden unter solchen Umstiinden matt und 
magern ab und bei den geringsten Veranlassungen treten Knochenbriiche 
ein. In mancheji Gegenden mit kalkarmen Boden zeigt sich in Folge des 
kalkarmen Grascs die Knochenbriichigkeit sehr hiiufig beim Rind, welches 
auf diesc Nahrung angewiesen ist, Katsuyaina hat gefundcn. dass 
Kaninchen beim Hungern anfangs etvvas weniger Kalk ausscheiden, die 
Menge steigt aber vom vierzehnten Ilungertage an langsam bis zum Tode. 
Die Magnesiawerthe dagegen zeigen ein continuirliches Sinken. Gerhart 
und ScJilcsiiiger fanden, dass die Ausscheidung des Kalks im Harne beim 
Gesunden und beim Diabetiker der Ammoniakauscheidung parallel geht 
und wie diesc durch Alkalizufuhr herabdriickbar ist. Unter abnormaler 
Siiurebildung 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 Entwickclungsperiode der Rachitis statt. 
Geloste Kalkverbindungen sind ferner nach Halliburton fiir die Herzthatig- 
keit von grosser Bedeutung, was von Jaques Loeb bestiitigt wurdc. Es ist 
daher von grosstcm Nachtheil fiir die regelmiissige Herzthiltigkeit, wenn 
der Kalk des Biutserums durch Fluornatrium oder oxalsaures Natron^ in 



* Zcilsclnift f. pliyi<iol. Clicmic 26, S. 542. 

' I'luoniatiiinu liat iilnigcns audi nocli audio \\ irkun_^cii, die iiiclit aul' der Kalkfalluiig bcruhen. 



1 



Ueber die Vertheiliiiig: des Kalks iiii thierischeii Org-aiiismus. 



145 



unlosliche Verbindungen iibergefuhrt wird. In neuester Zeit hat Fricden- 
t/ial^ es sehr wahrschcinlich gemacht, dass die Giftwirkung der Seifen 
(oelsaures Natron) bei Injection in das Blut auf der kalkfiillenden Wirkung 
derselben beruht. Er sagt richtig : ,.wie liiitte man vermiitlien konnen, 
dass eine Substanz, die in grossen Dosen verfiittert werden kann fast wic 
ein Nahrungsmittel, die in kleinen Mengen einen normalen Bestandtheil 
der Gewebc und des Blutes bildet, in die Blutbahn eingefiihrt schon in 
Dosen von 0,1 g. pro Kilo Thier den Tod im Augenblick der Einfiihrung 
herbeifiihren konne. Mnnk hat gezeigt, dass es nicht etwa das aus den 
Seifen freiwcrdende Natron ist, welches die Giftwirkung bedingt. Dieses 
wiirde ja auch bei dem Kohlensilure gchalt der l-Jiutes sofort in Carbonat 
ubergehen. 

Obwohl nun jedc thierische Zelle Kalksalze benothigt, so existirte 
docli bis in die neuere Zeit herein keine befriedigende Theorie der Funk- 
tionen des Kalks fiir die thierischen Zellen. Auch Pflanzen niit Ausnahme 
der niedersten Formen bediirfen des Kalks. Was nun diese Funktion des 
Kalks in den Pflanzen betrifft, so hat O. Loezu aus der Giftwirkung des 
neutralen oxalsauren Kalis, welche er unter dem Mikroskop verfolgte, 
geschlossen, dass sowohl die Chlorophyllkorper als auch der Zellkern aus 
Kalkvcrbindemgen von Nucleoproteiden aufgebaut sind. Die Funktion 
(Icr Magnesia in den Pflanzen dagegen bestehen nach ihm- in der Er- 
moglichung der Assimilation der Phosphorsilure bei dor Bildung von 
Lecithin und Nucleoproteiden, weil die Phosphorsaurc ani leichteslen von 
alien in den Pflanzen sich findenden Phosphaten aus dem sekundarcn 
Magncsiumphosphat abgespalten werden kann. Er hattc die merkwiirdige 
Giftwirkung von oxalsauren Salzen, wilche nur fiir hohere Thiere untl 
einige Pflanzen bckannt war, nicht nur bei verschiedenen Phaneiogamen,-'- 
sondern bei den hoheren Algen und niedersten Thieren beobachtet. In 
des Wirkung der Oxalate auf nicdere Wasserthiere licssen sich nun grosse 
IJnterschiede erkenncn, der Tod trat bei einigen yXrten (Asseln, Copepo- 



• Arcliiv fiir Aiuilomif iiiul I'lusioldt^io looi. 
•-' Flora 1892, 36S-394. 

3 \'on Intcresse ist bier die Tliatsadio. <!as> joilixli t;erini;i- Mention lii-^liclicr (Oxalate nornialorw else 
II manclien PflaiiztMi vinkommen konnen. 



146 M. Toyoiiaga: 

den, Rotatorien) sehr bald, bei anderen (Wasserkaferii, Wassermilben und 
Nematoden) aber weit spiiter ein, was wohl mit der verschiedeiien 
Schnelligkeit zusammenhangen mag-, mit der jene Salze zu den wichtigeren 
Organen vordringen konnen. 

In 0.5 proc. Losungen neutralen Kalium- oder Nalrium-Oxalats sind 
Asseln, Copepoden und Rotatorien in 30-50 Minuten todt, dann folgen 
Egel und Planarien, liierauf Insektenlarven und Ostracoden, wilhrend nacli 
24 Stunden noch leben Wasserkiifer, 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 ciner o.i proc. Losung 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. 

Fadcnlagen, wie Zyg7icinn^ JlFoiigeotia, J^anchcria, SpJiaeroplca, Cla- 
(lopliora, Oedogoiiiuni sterben binnen 24 Stunden unter Verquellung der 
Chlorophyllkorper in einer 0.5 proc. Losung von neutralem oxalsaurem 
Kali ab. Bei Spirogyren lilsst sich sehr gut beobachten, dass zuerst der 
Zellkern angegriffen wird. Derselbe quillt in einer 0.5 proc. Losung nach 
ciniger Zcit auf und wird ofter zu einem unregelmiissigen zackigen Gebilde. 
Lilsst man aber eine 2 proc. Losung auf diese Algen cinwirken, so gewahrt 
man schon nach 5 Minuten, dass die Kerne sich auffallend stark contra- 
hiren und nach 10 Minuten kcin einziger Kern mehr intact geblieben ist. 
I), IS C_\topIasma ist allcm Anschein nach noch \ollig unverletzt, doch 
crholen sich die Zellcn nicht wieder, wenn sie nach 10 Minuten wieder in 
kalkhaltiges Ouellwasser zuriickversetzt werden, die Zellen sind nach 24 
Stur.den in alien Theilen abgestorbcn. Der I^influss der 2 proc. Oxalat- 
l<')sung macht sich bei den Chlorophyllbandern der Spirogyren \w ca. 30 
iMinuten gcltcnd, wobci einc WMiindcrung der Conturen duich Verqueli- 
uncr sichtbar wird.- 



' Wasscnnillicn slarKc-n nsl iiaili 20-22 SUhkIl'H in einer i proc. l.iisunii oxalsauren Natrons. 
• I!ei Controlversuflien mit eiiensn slarKen I .("isnnLjeii vdii sclnvcfcisanrcin mlcr wcinsam'eni Kai 
liilicn jene I''rcl)cinunji[cn aus. 



I 



Ueber die Vortlioilimer <lcs Ksilks im thicrisolieii Org-aiiismiis. jaj 

Von WiclUigkeit ist es, bei der Thierfiitterung solche Pfianzen als 
Nalirung auszuscliliessen, wclchc losliche oxalsaure Sal;ce in geringen 
Mengen enthalten ; cs wircl einerseits der losliclie Kalk im Verdauungs- 
tractus ausgefiillt und durch den nnangelliaften Kalkgclialt entwickelt sich 
Knockenbriichigkeit und andererseits treten durch Resorption ins Blut 
andre krankhafte Erscheinungen auf, die raschen Verfali und Tod herbei- 
fiihren. In den RiibenbUlttern ist z.B. die Oxalsaure theils an Natron, 
theils an Kalk gebunden. Als Endrcsultat vieler Versuche ergab sich : 
Oxalsiiurc cnthaltendes Futter ist in geringer Menge, wenn cs nur kur/.e 
Zeit gegeben wird, als unschiidlich auzusehen. Werden jedoch die 
Bedingungen fiir die Unschildlichkeit des Putters (Zusatz von Calcium- 
carbonat) nicht erfiillt, so entwickeln sich die Symptome der chronischen 
Oxalsaure- Vergiftung, wobei zuniichst die Nieren und Knochen in Mit- 
leldenschaft gezogen werden. 

Diese allgemeine Giftwirkung ncutralcr oxalsaurer Salze bei niederen, 
sowohl wie hoheren Tiiicren erklart sich am einfachsten, wenn wir die bei 
Pfianzen sich ergebenden Schliisse, dass die Zellkerne zu ihrer Organi- 
sation Kalkvcrbindungen von Nucleoproteiden bediirfen/ audi auf die 
thierischen Zellen anwenden. Loe7a hat den Satz aufgcstellt : ,Je grosser 
die Zellkernmasse in einem Organ, desto mehr Kalk enthiilt sie." Dieser 
Satz hat aber noch keine Beriicksichtigung bei den Physiologcn gefimden. 
Vergleicht man jedoch bei verschiedenen thierischen Organen den Kalk- 
gehalt mit der relativen Grossc der Zellkerne, so findet man eine aulTallendc 
Uebereinstimmung mit jener Folgerung, soweit die bis jetzt vorlicgcnden 
Analysen Material zu Vergleichcn liefern. Leider sind aber manchc 
Organe nocli gar nicht im Bezug auf ihrc .Mineral bestandtheile quar.titativ 
untersucht, wie z. B. die Lunge, die peripheren Nerven, die Nebcnnierc, 
die Speicheldriisen und die Iloden. Von einigen Organen sind erst in 
neucster Zeit Aschenanalysen bekannt geworden. 

Aus dcm obigen Satz wiirde z. B. folgen, dass Driiscn kalkreicher scin 
miissen, als Muskeln, weil sic grosscrc Zelllccrnc haben. ferner dass 
Muskcln nicdercr Thierc mchr Kalk cntlialtcn. als die der Warmbliiter. 

* AiisijenriiniiK'n >-inil luir illo nii'<Ii'r-~li'n Al^i'u- uiiil I'il/lnrnuni. 



148 



31. Tovou«8:a : 



da erstere ebenfalls grossere Zellkernc besitzen. Locw^ liat einige von 

Katrj- vor einigen Jahren publiciertc Aiialysen dcs Muskelfleisches hoherer 

und niederer Thiere verglichen sowohl iinter sich, als audi mit dem 

vorliegenden Analysenmaterial von Driiseii uiid fand seine Folgcrung 

bcstiitigt. Wir wollen im Folgenden siimmtliche Analyscn von Kaiz 

beriicksichtigen. Dleser Autor fand 

ill looo Til. frischen Muskel.s von Warmbliitern : 

Calcium. 
... 0,0748 

... 0,0806 



Mensclienfleisch 
Schweinflei.scli ... 

RindPieisch 

Kalbfleisch 

Hirschfleiscii 
Kaninchenfleisch 
Hundefleisch 
Katzenfleisch 
Hiihnerfleisch ... 

Durcliscliiiitt 



0,0211 
0,1444 
0,0959 
0.1832 
0,0685 
0,0846 
0,1051 

0,0954 



Diese Zalil ist ctwas luiher als die von Ihnigc"" schon fiiiher gefundenen 

Zahlen 0,086 und 0,072 CaO, ent.sprechend o,o6r und 0,053 Ca. Anderer- 

seits fand Oidtviaiui in dtr Leber, dieser grossten Driise der Siiugethiere, 

0,284 Theile Calcium in 1000 Thcilen des Organs oder nahe 3} mal so vicl 

als jener Durchschnittgehalt beim Muskcl betrilgt. Bei Kaltbliitern fand 

Katr: in 1000 Tlieilcn Muskcl : 

Calcium. 
0,1566 



iK'im 1^^-oscli... 
.Scliclinsch 
Aal ... 
If<^cht ... 



Duiclisclinitt 



0,2202 
0.3913 
0,3977 

0.2913 



' 1 lie pliy.-iolci;. RdIc i>I mihci-.il .Null ii-iil<. I'.ulli-tiu No. iS. V . S. 1 )i-p;irtnR'iit i.l' Ai;iiciilture, 
\\':i>-liinj^t(in 1S99. 

- Jiilircsbcridit 1. riiiciclieiuie 1S96, p. 479. 
* Zfitschrift r. iiIiysiol(ic(. (.'licmic 9, p. 60. 



I 



Ueber die Vertlieiluiig des Kalk«^ im thierisoheu OrgaiiisinuK. 



'49 



Wir findeii somit, class der Kalkgehalt der Muskein von Batracliicrn iind 

Fischen weit grosser ist als der bei den Muskein von vSiiugethieren, iti 

Uebereinstimmung- mit obiger Folgerung. Fur den Magiiesiumgehalt 

ergiebt sich umgekehrt, dass derselbe bei den Muskein von Saugethieren 

grosser ist als bei denen von Batrachiern und F'ischen ; namlich 

in looo Th. frischer Substanz : 

Magnesium 
... o,2ii6 

... 0,2823 



Menschenfleisch 
Sell weinflei sell ... 

Rindfleisch 

Kalbfleisch 

Hirschfleisch 

Kaninchenfleisch 

Hundefleisch 

Katzenfleisch 

Huhnerfleisch 

Durchschnitt 



0,2434 
0,3044 
0,2906 
0,2869 
0,2370 
0,2863 
0,3713 



0,2793 

In Uebereinstimmung damit fand schon Bunge fiir 1000 Theile Fleisch 
0,412 und 0,381 Magnesia, oder 0,249 ""^1 0,230 Magnesium. Vergleichen 
wir hiermit den von Kat.':^ gefundenen Magnesiagehalt der Muskein von 
Batrachiern und Fischen, namlich : 

In 1000 Theilen : 



Froschfleisch 
Schellfishfleisch 

Aalfleisch 

Hechtfleisch 

Durchschnitt 



Magnesium 
• 0,2353 

. 0,1670 

,. 0,1782 

. 0,3102 

,. 0,2227 



so sehen wir, dass der Magnesiagehalt liicr geringcr ist als dort, ucnn 
auch der Unterschied fur Magnesium gcringer ist als der gegentheilige fiir 
Calcium. (9/V//;//^r;/;/' fand auf 3,62 Theile Kalk in der Leber der Siiugc- 
thiere nur 0,19 Theile Magnesia, oder auf 0,284 Theile Calcium in 1000 



* Ciliort von Halliluirtoii, S. 53S seiner cliien'ii-;chcn Pliysioloyie. 



150 



M. Toyouaga : 



Theil Organ nur 0,017 Theile Magnesium. Es ist somit der Kalkgehalt 

weit grosser als der Magnesiagehalt, wahrend beiin Muskel uingekehrt der 

Kalkgehalt geringer ist als der Magnesiagehalt. Gossiiiann^ fand iihn- 

liche Zahlen fur die Pancreasdriise und Niere wic Oidtmann fiir die Leber. 

In 1000 Theilen Organ sind cnthalten : 

Pancreas 
I Ca 0,2451 

'Mg 0,0383 



Rind ... 

Mcnsch 



Niere 
0,1184 



0,0410 

Ca 0,3958 0,2008 

/ Mg 0,0833 0,0472 

Seither sind wciterc Analysen einigcr thierischcr (.)rganc erschienen. So 
hat Lulling die anorganischen Bestandtheilc der Pancreasdriise von zvvei 
alten Frauen (|uantitativ bestimnit und fand 2,^6% Calcium und 1,48^ 
Magnesium (wahrscheinlich in 100 Theilen- der Asche). Ribant^ 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 Organc. 1m- erhielt folgende Werthe auf 
Trockensubstanz bezogen : 

Magnesium. 

IH. I. II. 

0/ II • 

0,141 0,054 0,05s 0,055 -08 2,62 2,56 
0.15S 0,070 0,083 0,067 3-5' 2,24 2,36 
0.098 0,026 0,025 0,023 ''7^* 4J3-4 4'26 



Calcium. 

I. H. 

(1/ 0/ 

Gan/t .Mi!/ 0,129 0,153 
I'ulj).) 0,247 0.183 

liiiKlcgc\vcl)i.' 0,046 0,108 



111. 



Ca : Mg. 
II. in. 



Audi A/or^ hat gefunden, dass in Miiz, Pancreas und Niere, ferner in 
Knorpcl und Bindegewebe der Kalkgehalt grosser ist als der Magnesia- 
uehalt. Derselbe fand, dass in Gehirn und Muskel das Vcrhidtnis -=r-^ 
kleiner als i ist, widnend cr fiir Milz, Pancreas und Niere die VerhiUtnisse 
fand 6.79; 4.05; und 1.84. ^l/ov scheint keine Kenntnis von den oben 
bereits discuticrten Beziehungen zwischcn Kernmasse und Kalkgehalt 



* JiiluX'sl)Ciiicht 1. 'riiiiTclu'iiiii.- 30. S. .(K). 
' Ibid. S, 386. 

•■' Ibid. S. 492. 

* JaliresbcriclU f. riiicrchcmic 30, S. 492. 



UelMT (lie Verthcilnisg dcs Kalks im Ihiei'ischoii Ori|?aui)^uius. 



i^i 



gehabt zu liabcii, soiist ware cr voii seincn Rcsultateii wcnigcr iiberrascht 
word en. 

Was meinc cigcnc Untersuclumg betrifft, so liabc ich zunachst die 
weisse und graue Gehirnsubstanz scparat auf Kalk- und Magnesiagehalt 
untersucht. Diese beideii Gehinisubstaiizcii unterscheiden sich bekannt- 
lich sehr bedeutend in mehreren Bczieliungen. Die grauc Substanz bcstcht 
aus Nervenzellcn mit eineni wohl entwickcltcn Nucleus, wilhrend die 
weisse Substanz ledigh'ch aus Nervenfibrillen besteht, welchc aus Nerven- 
zellcn hervorgehen und nicht wie diesc wolil ausgebildetc Kerne haben. 
Obuohl zahlreiche Untersucliungen beider Substanzen vorliegen, was die 
organisclien Restandtheile betrifft, existirt docli noch kcine Analyse der 
Asche von jeder dieser Substanzen fiir sich. Trotzdcni mussten solchc 
Analyscn von hoheni Werth scin, mit Riicksicht auf die grosse ZcUkern- 
masse der grauen Substanz. ]3cr totalc Aschcngehalt dcs Gchirns variicrt 
nacli vcrschicdcnen Autorcn von o, i zu i,oj^'q, und nach GcogJichan^ 
enthalten looo Theilc Totril- (jehirn : 

(Jaii/c A^elif 
2,95-7,08 

Wilhrend Gcoghcha)i das Lccitln'n und damit cine belrachtlichc Mengc 
Phosphorsilure vor dem ICiniischcrn des Ilirns init Aether entfernte, soniit 
audi weniger Totalasche erhielt als anderc Autorcn, fiihrt auch das 
dirccte Eiiiasclicrn dcs Hirns zu Fehlerquellcn, da vicl Phosphorsaure aus 
dcni Lecithin frei wird, die nun wahrend des l^'nascherns mit der gliihcn- 
den kohligen Masse zu langc in Beriihrung bleibt.'-' 'Icli habc. urn 
Vcrlustc zu vcrmciden, und den Verbrcnnungsprocess zu erlcichtcrn, bci 
dem I'Linilschcrn cine bestimmte Mengc Soda (meist 5 g.) zugesctzt und 
die ?ilenge nach dem Veraschen und Wiegcn wieder abgczogen. Allcin 
auch diese Methode liefcrt keine fehlerfreic Bestimmung der Gesammt- 



K 


\.. 




Mk' 


(■;i 


CI 


^''\ 


CO, 


i-c(ro,\. 


0,5^-1.77 . 


045-1 


)i • 


0,017-0,072 


0,006-0,022 


0,43-1-32 


0.95-2,01 


0,24-0,79 


0,01-0,09 



1 Ci(irl in //(>///7'//;/(»//'.v Clioniical I'hysioloi^y iSoi, i\ 517. Xadi Z. 1". physiul. Chcui. IVl. i. 

^- 335- 

~ Cico^JicluDi ZL'iolo iihiij^cns, (1,l^s (lic>o iVcic l'l\osphoi>aiiio auch Carbonalc /crsctzt, wclcho 
In'ini I'ina'sdioni dcs Goliinis orliallcn woixlcn. I'.iii von Lecithin bcdVeitcs Cchiin liclcrt n.'iiulich nach 
hni cine Kohloiis.'imv haHiu;o Aschc. 



1^2 



31. Toyouagii: 



asclic, well wahrend dcs Veraschens, eiii Thcil der zugestztcn Soda in 
Folge der Bindung der Lecithinphosphorsaurc Kohlensaure verliert. 
Doch niir kam es ledfglicli auf die Bestimmuiig" der Kalk- und Magncsia- 
inengen fiir lOOO Theile frischen Gehirnes an und ich babe dessbalb das 
I'robleni, eine mogliclist cinwandfreie Bestinimung uer Gesamtmineral- 
stofife im Geliirn zu liefern, noch einstweilen bei Seite gclassen. 

Ich habe sowolil das Gehirn vom Pferde als aucli vom Kalb unter- 
sucht, nachdem ich die weisse Substanz so gut als moglich v-mi der grauen 
trennteJ Eine frisch abgewogene Portion wurde zunachst im Wasserbade 
eingetrocknet, zuletzt im Luftbadc. Nacli Zusatz von Soda wurde ein- 
geiischert, nach Extraction mit Wasser die Asche weissgebrannt und nun 
Kaik und Magnesia in iibh'cher Weise bestimmt. In gleichcr Weisc 
bestiinmtc icli ferner diese Basen in den pcriphcren Nerven und der Lunge 
des Pferdes. Die Resultate sind aus folcrcnder Tabelle crsichtlich. 







Ant;;ewand- 

tcs Gewicbt 

an frischer 

Substanz. 

g- 


Asdic 


Kalk. 


Magnesia. 




In der 
Asclic 


In 1000 

Till. frischer 

Substanz. 


In der 
Asche 


In 1000 

Till, frischer 
Sulistanz. 


Kail,- 


Grauc Ilirnsubstanz. 


89,50 


0,8440 


0,0329 


0.368 


0,0227 


0,254 


Weisse Ilirnsubstanz. 


158,00 


1,3918 


0,0092 


0,058 


0,0094 


0,060 


T'fcixl- 


Grauc Ilirnsubstanz. 


23.970 


0,2794 


0,0261 


1,089 


0,01 1 1 


0,463 


Weisse Ilirnsubstanz. 


68,106 


1,0350 


0,0045 


0,052 


0,0138 


0,203 


rcriplKTc Xcivcn dcs rfcrdc«. 


39,720 


0,20I I 


0,0315 


0,794 


0,0239 


0,602 


Luntrc dcs Pfcrdcs. 


61,050 


0,2035 


0,0313 


0,513 


0,0274 


0,449 



l^s hat somit die Analyse in Analogic mit den obcnerwahntcn Vcrhalt- 
nissen audi hier fi'ir die grauc Substanz mit ilircn zahlreichcn Zcllkerncn 
cincn grosscrcn Kalkgehalt crgcben als fiir die weisse, an Zellkcrncn wcit 



• Die grauc Sub>l,iii/ macht 37.7-39.0%', die wcis^c 61.0-62. 3_?o dc> Gcsanimthirns aus. 



lloJjor dio Vortlioiliiiif>' dcs Kalk^ iiii tliiorisclifii Orf^aiusmus. j-^ 

iirnicrc^. Feriicr iiberwicf^t in der graiien Substanz dcr Kalkijchalt i'lbcr 
den IMagncsiagehalt, wiilircnd in dcr weissen das Umgckelirtc dcr Fall 
ist. Dcr Nucleingchalt fiir die graue und weisse Substanz ist noch nicht 
separat cliemisch bestimmt wordcn. Xach Jaksch cnthiilt dcs Totalhirn in 
lOOO Thcilcn nur 3 Thcilc Nuclcin, nach GeogJieiian g^ar nur 1,34-1,62. 
Wenn nun auch die Grundlage dieser Bercchnungen kcinc ganz sichere 
ist, so bleibt sovicl jcdenfalls richtig, dass der Nucleingchalt nur gering ist. 
Uamit vviirde also auch der relativ geringe Kalkgchalt des Totalhirns 
stinimen. Doch diirfte inimerhin die von Geoghchan hicfiir gefundene 
Zahl 0,006 Ca auf 1000 Tlieile Gesammthirn auf einer mit FcliKrn bchafte- 
tcn Analyse beruhen. 

Magnesia und Kalkgchalt des Gehirns scheincn betriichtlichen 
Schwankungen unterworfen zu scin; wahrscheinlich iiben Art und y\lter dcr 
Thiere, sowie dcr wcclisclnde l'"ett- und Wassergehalf'^ hicrauf, wie auf den 
Procentsatz an Gesammtasche cinen wescntlichcn Einfluss aus. Wie schon 
erwiUmt, fanden verschiedenc Autoren den Aschegchalt dcs Gesammthirns 
zu o.i-i.o Procent ; ferner wurde das specifisclie Geuicht der grauen 
Substanz von 1,029-1.039 schwankend gefunden, Bei krankhaften Zustiln- 
den ferner warden welt grossere Schwankungen eintreten konnen. Um nur 
ein Beispiel zu erwiihnen, kann bei Erkrankungen von Blutgefiissen (Aorta) 
der Kalkgchalt bis zu dem 20 fachen des normalcn ansteigen.-' 



' Audi wcnn w ir doii vcrichiedfiion I'VU- uiul W'asscrtjclKxlt I)L'iucksii-1Uii;on, MciM ilic>o> ricliliij. 
Ks LTijilit sicli I'iJr 100 'i'lioiio Ircclviic tcltfivic i^rauo Sul'stanz hcim Kalh =0.290 Tlil. (.'aO; ilitto 
wcissc Sulistanz 0.075 '^'^'' ^'i^^- 

2 In I/aniiuarsti-n's I.chrbuch dor pbysiologisclien Chemio, 111 .Vuflaijo, S. 353. lielsst fs : ..die 
Moni^c (k's Wasscvj? ini Ccliiin ist grosser liei jungcrn Individiien nis I'oi l\r\vacliseiien." Wie aus der 
daraiit" lolgcndon 'I'alicllc (.■rsichtlicli ist, gil>t cs Ausiialnuoii. 

3 (/(7;<7/, Jalircslicriclit IQr riuorclicmio, 1000. S. i;ii. 



1 54 ^^' Toyoiiaaa : I'oher dio Veiiheilmig des Kalks im tliierischen Oi'franismu)*. 



Zusammenfassung. 

In Ucbcrcinstimnnin,q; mit der von O. Loeiv g-ezogencn Folgerung^, 
class dcr Kalkgehalt mit der Masse der Zellkcrnc wiichst, stclit das 
Resultat mein.er Untersuchung, dass die graiie Ilirnsubstanz relativ 
kalkreiclier ist, als die weissc. 



On the Digestive Power of the hitestiiial Canal. 

P.Y 

S. Sa-wamura. 



After the experiments of TJiiry ■\\\(\ Quince the digestive function of the 
main part of tlie lower intestins was regarded to be of minor iniportance, 
but recent investigations disproved this view. Besides the accelerating 
action on th.e enzyms of the pancreatic juice observed by Schepozvalnikoiv'^ 
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 Brown and Heron,'^ in 
the small intestine of the dog hy Kruger,^ Grunert,-' and Schepoivaluikoii.','^ 
and in the coceum and colon of various animals by Litdwig Vclla.' On 
the other hand, an enzym acting on inulin was proved to be absent in 
the human intestinal juice by Dcuiant.^ Sucrase was shown to exist in 
the human intestinal juice by Dcinant,'^ in the small intestines of the 
swine by Broivn and Heron,'' ^^ in the intestines of th.c rabbit by Pavy,'^'^ 
in the coecum and colon of various animals by Vella,'''- \\\ the small 
intestines of the dog by Grihiert,^'^ Krilger,^^ :\\\d Basfidtne/li^^'" and in 
the small intestines of man by Miura.^'^ Maltase was observed in the 
small intestines of the swine by Brown and Hcron,^' in the intestines of 

1 and c Jalirc>l)cricht I'iir Ticicliciiiie vol. XX, XI. p. 379. 

' 8 and !» il)id. vol. TX. p. 222. 

3 1" and >7 ii,i(i. vol. X. p. 77. 

* and 1* ibid. vol. XX. Mil. p. ;,;,7. 

^ and 13 ibid. vol. XXI. p. 27^ 

7 and 13 ibid. vol. X\'. p. 297. 
'^ ibid. vol. XIV. p. 294. 

1" Jalnesbencht fur Agrikultuivlicmio. 1S90. p. 523. 
'« Jahrt-~bericht I'vir 'l"i<.-icl\<'niir. \'ol. XX\'. p. 2SS. 



I c5 ^' Sjuvjinnirii: 

the rabbit by Ptn'y,^ in the cocciim and colon of various animals l)\' 
Vella," and in the small intcstiiics of dogs and cattle by Pant:" and 
Vogc'l ;''' Lactase, in the intestines of various young animals by RiJiiiaini 
a.\\d Lappv,'^ Pant.'::: and Vogel,'' Portier,'^ Orbair and JVciulaud.^ Pantr; 
and ]^ogeI'^ found an enzym which acted upon raffinose in the intestines 
of tlie dog and cattle. As to the occurrence of cytasc, the opinions 
differed, but it was finally decided in tlic negative. 

As to the behavior of digestive juices to various hemicelluloses, our 
knowledge is still imperfect. Hanber^^^ '\\\ 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 
gl>'cerin-extracts ol the stomach and pancreas of the same animal trans- 
formed mucilages into sugars, vvh.ilc pt}'alin had no such effect. Stoiic and 
joiies,^^ and Liiuiscy and Holland^- observed the digestibility of pentosan 
by the rabbit, Weiske^'' by the sheep and K'Cnig and ReinJiartd b\- 
man.^"* •* •"' But Nilsoji^^ observed that no sugar w^as 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 T'i'/Aj;^ " observed that it 
is only emulsified by them, wdiile the absence of a special enzym was 



1 j;ihrcsl)criclit I'Cir 'ricrclu'Uiio. vol. XIX". [i. 294. 

2 and *■'• Ditto, vol. W. p. 297. 

3 •■"> aii.l '■> Ditto, vol. XXI\'. p. 304. 
■» Ditto, vol. X.W. p. 2S6. 

6 Ditto, vol. XX. Vlir. p. 73;, 

' Ditto, vol. XX. 1\. p. 3S4. 

« Ditto, vol. XXIX. p. 382. 

*" Ditto, vol. IV. p. 375. 

^' laliif^licritht fill- .\L,a-ikiilturchcniii-. iS(\:;. 11.373. 
»2 anil '^ Ditto, vol. 1S95. p. 431. 

Ditto. 1893. p. 53. 
^^ ChciniFches Ccntnil-lUatt. 1902. vol. i. p. 673. 

*" I Icrliivora can (litest pentosan hctti-r than onmivora, and as it i> al\va\-^ livdratol liy di|_;('-.tion, 
there- must lic present a special kind of cytase in the intestines. 
Zeitschrift fiir j)hysiol. Chem. \'ol. 36. p .65-66. 
*' Ji^hresherieht fiir Tiercheniie. vol. X\'. p. 297. 



i>\\ the Dig-estivo Power of (lie lutestiiiiil riiiial. 157 

proved by Dcmant^ in the secretion of the intestinal tract, and by 
Sche powalnikcnc" in the intestines of the dog. A proteolytic enzym was 
proved to be present in the intestinal secretion of the dog by Grilnert^ 
GacJict,^ and Schepoivalnikozv^' bnt DeinaiW^' (in the hnman intestines) 
and Kr'ugcr'' (in the intestines of the dog) questioned its i)rcsence. 

Although various kinds of enzyms were hitherto observed in the 
intestines, little attention has been paid to the special parts of the 
intestines in wlu'ch they are produced. Since both, Licberkului s and 
Britniicr 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 
mannaii and galactan in the different parts of the intestines. In Japan a 
preparation consisting chiefly of mannan and derived from the root of 
ConopJialliiS Konyaku and further the common agar are consumed generally 
by the people and since those hemicelluloses arc digested we have to 
infer the occurrence of mannasc and galactasc 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 for a day. '^o grs. uf the 
small intestines, coecum and colon cut into pieces, were digested with 
about five times their weight of dilute alcohol (35,^0) for a week. The 
filtered alcohol extracts were precipitated with ether-alcohol, the prccii)itatc 

aiul " Jalucsbciicht liir Tiorchoinic vol. IX. p. 2.2.1. 
2 and •'■ Ditto, vol. XX. IX. p. 379. 
" Ditto, vol. XXI. p. 273. 

•» Ditto, vol. XX, VII. p. 377. 

7 Ditto, vol, XXVIII. p. 357. 

" Ellcu^cr. I Ii:-to!ogio tier Ilaussaugclhicrc. p. 694. 



iss 



S. Sawamura 



dissolved in a o.2\% sohition of sodiuni carbonate, and with the addition 
of sonic thymol served for the following experiments with crude fibrin, 
neutral olive oil, starch s()liiti(Mi, cane-sugar, mannan and cellulose (filter- 
paper). The results observed after three day's digestion at 36"C were as 
follows : — 



-Matcri.ils u.'^cd 


J'lxtract from tlic small 
intestines 


ICxtr.ict from tlie coccum 


]''.xlract from the cok>n 


I'ihrin 


A little aUackeJ 


Not dissolved 


Xot dissolval 


Oil 


No acid reaction 


Xo acid reaction 


X"o acid reaction 


Starch 


Much sugar 


Some sugar 


Little sugar 


Afaniian 


SugaV reaction j ositive 


Sugar reaction positive 


Sugar reactioji positive 


Cancsufjar 


Lnertctl 


Xot inverted 


Xol inverted 


Cellulose 


\o sugar 


Xo sugar 


X'o sugar 



The itlcntity ol the sugar produced with mannosc was proved by the 
production of the difhctdtly soluble characteristic phenylln'drazon. 

The enzym-production is therefore not quite the saiiie in the various 
parts uf the intestines. As to the en7.}-ni which acts upon mannan, — the 
mannase — it exists in all three divisions of the intestines. 

A second exiieriment was perfornied with the intestines of .i swine. 
The extracts were prepared by digesting res[KCtively 50 grs. of the 
duodenum, coi cum., colon and pancreas (as a control) in 200 cc. ol 20_^q' 
alcohol for a weelc. 'Ihe exiKriments were carried on with the addition 
of some lh\niol. The results observed after digestion at 3OC. lor three 
(.lays were as (oliows': — 



' 'I'iie extract was proved to Ik- Iree from any reducing sugars Uy testing with I-i/i/ing's solution. 



Oil Digestive To^ver of l!ie {ulestiiiiii CiiiuiL 



'59 





Mxlract from tlie 
duodenum. 


I'xtract from tlie 
coecuni. 


ICxtract from the 
co!o!). 


Extract from the 
pancreas. 


l*'il)l-ill 


Not 

attacked 


Not 

attacked 


Not 
attackcfl 


Dissolved 
conifilctcty 


Oil 


Xo acid 
reaction 


X'o acid 
reaction 


X'o acid 
reaction 


Acid 
reaction 


Stai-cli 


Sut^ar reaction 
positive 


Sugar reaction 
positive 


Sugar i-eaction 
positive 


Sugar reaction 
positive 


^[a^nan 


Sui^ar reaction 
positive 


Sugar reaction 
positive 


Sugar reaction 
positive 


Sugar reacnion 
positive 


("■alactan 


Su<j;ar reaction 
negative 


Sugai" reaction 
negative 


Sugar reaction 
negative 


Sugar reaction 
negative 


Saccliarose 


Inverted 


Xot inverted 


X'ot inverted 


X'ot iiiverted 



The sugar produced froivi mannan was proved to be maunosc by test- 
ing; with plienylliydrazinc. 

As will be seen in the above table proteolytic and lipatic enzyms 
were absent in the intestines, while diastase ar.d niannase v^-ere present 
in all the parts of the intestines.* 

Contrary to our expectation the absence of qalactase was proved in 
this experiment.- We may conclude from th.ese result as follows: — 

1. In the intestines and pancreas of the higher animals, there is 
present inaniicisc besides the other er.zyms already obser\'ed. 

2. Tlie enzym-production is ditlercnt in the small intestines, coecum 
and colon, the most notable being the absence of sitcrasc in Ciecuni and 
colon in the case of the horse and swine, and o{ trypsin in the c.ise if the 
horse. 



' Diastase was also present in the rectum. 

2 .\ mucilage consisting ot' mannan, galactan, and arahan w.is digestal wiih the extract ot the 
inte.-^tincs of the horse and swine, and it was fourid that some arabinose was formcvl in both these cases, 
as was proved l>v phlorogluein and hydrochloric acid in the alcoholic extnicts of the digesto.! mucilage. 



i6o S. Sawamiira: On Digestive Powor of the lutestiiial Caual. 

3 Cytase was not observed in this experiment. 

4 The function of the intestines in digestion nuist be regarded to be 
very important, since they secrete enzyms such as sucrase that arc not 
contained in tlie pancreatic juice. 



On the Action of Manganese Compounds on Plants. 

BV 
O. lioe-w and S. Sa-wa. 



With riate XII. 

The ahiiost univcrpal occurrence of manganese in the aslies of plants 
is a fact known long since, but as plants have been raised in water-culture 
to perfection in absence of manganese, this clement 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^ MugO^ and only 1,07^ 
Fe^Os. ^ 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 leaves are especially rich in it.^ 

It deserves to be pointed out that in one case mangano manganic oxid, 
Mu;, Oi, formed even the chief constituent of a plant ash. This being really 
an extraordinary case we mention the composition of the said plant ash. 
7\ ScJir'oder'^ found on analysis of the ash of various [\arts of a pine tree 
{Ptniis abics,) among other things the following result : 

Leaves Bark 

Pure ash 3-0640^^ i-'^os %' 

In 100 parts of pure ash : 

K.O 14.480^ 20.46% 

Na.jO 0.67 0.38 

CaO I \..\2 '4-7- 

i U'o/ff's Tables of riant Ashes. I. \\ m. 

* /\tii>iann found it i:i the pollen ijrains of tlio imiio, liotan. Contni!l>l. iSoS. 

* PiiiiarJ, Qo\\\\\. rend ; vol 126, p. 550. 

* Korstcheinische uiul pllan/enphysiolosrisclie I'litorsuchungon, Tliaraini, 1:^78. Also : l.ihn^bc 
liclit fur AijriculturcluMnie 1S7S. 



MgO 8.46 7. 14 

Fe^ O;; 4.04 3.61 

■ Mil, 0, 35.53 41.23 

V>0, 9-59 6/3 

SO, 8.68 2.69 

SiO, 6.33 3.04 

1 he manganese content, as Mn, Oj, for tlic dr\' matter of these leaves 
was therefore J.oS% and that of the bark 0.66 j)^, corrcsi)on(h"nq- to O.69 
and o.4;zP(^]\In respectively. — Animal organs contain much less manganese 
than vegetable organs. Richc found only 0.$ milligrams IMn, O^ in one 
kilo of blood and according to other authors it is often completely absent. 
I[7/r,cr/' (1833) observed it in the ash of the l:\'er and teeth, Weideubuscli 
in th.c bile, Tlorsford (1851) in urine, Pollacci (1871) in milk and eggs, 
Ulaiivieiie (1883) in hairs and bones, PicJiard (1898) \\\ molluscs, crabs, 
sardines, pigs blood and hens eggs. The general occurrence in animals 
forms apparently a contrast to t!ie highly poisonous properties it shows on 
subcutaneous and. intravenous injections. Eight m.illigrams of manganous 
oxid in the form of sodium-manganese citrate represents according to Kobcrt 
the letal dose per kilo body weight of a dog. On introduction into the 
stomach even large doses of manganese compounds jjrovc harmless, on 
account of deficient absorption by the intestinal walls. ^ 

In regard to the behavior of plants towards manganese compounds, 
but few experiments ha\'e been iiiade and these show, that manganese 
cannot replace the related iron in regard to the production of chlorophyll, 
and (liat manganous and manganic phosphate suspentled in culture 
solutions can exert an injurious effect." Recently GigUoW^ applied peroxid 
of manganese as an addition to various manures on fields and observed in 



1 In re.cnt times manganese com} ounds commenced (o play a role in llierepeiilics. A mangancse- 
iron-pc'iitoi) preparation is frequently recommended for certain disorders. 

2 Cf, Kinur and I.inwuis, Landw. ycrsuchs-Stat. vol. 8, p. 12S and Wagner, il)i(l. vol. 13, p. 
69 and 278. 

3 Ann. Delia K. Scuola Sup. di rortici 1900. The exiicriments were made with wheat. The 
peroxid of manganese was applied in the j^roportion of 1,14 ctw. jier ha. Cf. also Centralhl. f. Agrieultur- 
chcmic 1902, Is'o. 3. 



On tlu' Action of iMang-ancsc ("oiiiimhuhIs oh riant>^. i(,o 

some cases a moderate increase, in otiiers a decrease of the harvest. The 
result was not decisive, wliich will jv>t create surprise since peroxid of 
manganese is a compound which hardly can be attacked and dissolved hy 
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 [)ropertics of these salts, so clost.-ly related to the salts of iron, 
considering further the occurrence of manganese in the ash of oxidizing 
eiizyrns {/>ertra//(/) and in that of certain nucleoproteids {Asd).^ 

In order to obscrvi the character of the injuries caused by manganese, 
young pea plants, 16-17 cm. high were placed in a solution oi o_2^^q 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 n.o 
trace of new rootlets became visible. The control plants, however, remain- 
ed perfecth' 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 in a 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. On the ninth day further observation was given 
up and comparative tests were made as to the color reactions upon o.xidiz- 
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 tiie 
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 fi\e drops of a 2^q alcoholic guaiac solution added 

' Hull. College of Agriculture, Tokyo ; vol, 4 Xo. 3. 

* Such d;>ta icier in this article to the anh\\lrous coihiouikI, not to the crysialli;:eil salt. 



164 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 t^uaiac 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 jjcroxid. One cc. of the filtrate (f) was diluted 
with 20 cc. of water and five drops of a 1% 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 weaky 
coloration was slowly developed. A colorometric test fifteen minutes after 
adding the reagents showed that in the latter case the 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 pcioxid was carried out. The 
green 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 tb.c oxidizing enzyms are 
more intense with the manganese plants than the control plants can 
give as also an account for the fading out of ihc green color of the leaves. 
Our result corroborates the statement of AV;7;v?av/' that the oxidizing 
enzyms act more [)o\vcrfully in presence of manganese com[)ounds tiian in 
their absence, and that iron compounds cannot produce an effect of the 
same intensity.^ The effect of the manganese seems to be the same, as an 
increase of the oxidizing enzyms by certain stimulants secreted by parasit- 
ary insects {ApJiides) and fungi. The )'ellow spots thus produced on 
leaves )ield according to Albert F. ll^oods more intense reactions upon 
oxidase antl peroxidase than an ecjual surface of the healthy leaves. 
Woods also observed a higher content of oxidases in etiolated shoots, than 
in normally green ones.^ 



^ Coiiiiit. ix-ml., vol. 124, l>. 1032. 

» 'HiLTc may exist cases in wliicli oxidi/inij; eii/yin-. are not a'^sociateil with inuiit^'anese or iron 
lut they will be less i^owerful in that case. ('f. also Saii/icii. Jnurn. I'liarin, Cheni. vol 1 1, p. 5^3 [iQOO-l 
3 C. 15ukt. II Abt. 5, S. 745 [1899.] 



On the Action of Mauganosc ConiiMmuds on plants. 15^ 

111 our next experiments' the nuiiiganous sulphate was applied in a 
mucli higher dilution in the expectation to diininish the injurious effects 
to a minimum. At the same time the mineral nutrients were offered to the 
plants in the following proportions ; 

Calcium nitrate 0.04^0 

Magnesium sulphate 0.01% 

Potassium nitrate 0-03/^ 

Monopotassium phosphate 0.02_%' 

Ammonium sulphate 0.01% 

To one portion was added o.oi ?o ferrous sulphate (control solution), to 
another o.02^q manganous sulphate and to a third O.oij^o ferrous sulphate 
plus o.02_^Q manganous sulphate. 

Shoots of barley and soy bean were placed in these solutions and 
kept near a window in a room whose temperatuie ranged from 4 — I2^C. 
during the first three weeks of observation. After a series of days the 
shoots in the solution containing manganese and iron jointly cxJiibited an 
increased gro7vtJi. The measurement revealed even considerable differences 
(see Table) ; gradually however these shoots turned rel/ozi'ish, their as- 
similation power was consequently de[)ressed and in further consequence 
of this the decreased nutrition led to a relaxation in the speed oi growth, 
as seen from the measurements of the soy bean plants. 

Experiment Avith Barley, 

Two shoots were placed in each of the solutions above mentioned 
IMarch 27. 

* \\c also liave m.nlc cxj'criiuculb with. a!t;ac Imt tlioy failed on accv.>unt of dcvclojinicnt o 
I'arasitcs, 



I 



1 66 



0. Locw mid S. Sjiv.a: 







Date of Measurement, 


cm. 


Increase after 
25 (lays. 

- 




March 27 


April 9 


AprH 22 


A 

Mil , 


34.0 


46.0 


51-5 


51-4 


IJ 


38.0 


47.2 


55-5 


46.5 


-Mn+l-V . 


A 

r. 


35-5 


59.0 


60.0 


71.S 




34-0 


51-6 


58.0 


70.6 


Fc- 


A 
B 


35-0 


43-4 


52.1 


4S.5 


35-0 


50-3 


57.7 


64.9 



Experiments with Soy bean. 
Three shoots were placed in eacli solution, ?klarch 25. 









1 )atc of Measurement 


, cm. 






March 25. 


April 8. 


April 22. 


April 30. 


May 10. 




A 

V> 
C 


9-3 


260 


37-0 


40.0 


45 -o 


Ml).- 


10.2 


22.0 


^54 


35-0 


40.5 




7.0 


22.5 


3 10 


31-6 


dead. 




1! 

c: 


9.6 


27.0 


40.0 


48.1 


56.5 


Mm + I'c.h 


9.S 


26.0 


402 


43-5 


43-5 




7-5 


235 


3S.2 


46.0 


51-5 




A 
1; 

C 

1 


7.8 


24.1 


36.5 


45.5 


56.5 


Fe.. 


9-3 


21.0 


30.0 


38.5 


45-0 




S.4 


2.J.2 


35-0 


43-5 


49.0 



Oil (he Action of 3Ianj?ajiesc Couipouuds on Plants. 167 

The gradual increase in tlie speed of [growth up to April 22 and the 
followini; diniiiuitioii of that si)ccd with the plants that received iron and 
manganese is still clearer seen from tiie average : 

March 25. April 8. April 22. April 30. May 10. 

Mn-fFe 8.9 25.3 39.4 45.8 50.5 cm. 

Fe 8.5 23.1 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 iiianganese in tiie cells is transformed 
into insoluble compounds^, a process, that probably takes place in nature, 
when [)lants absorb nianganese compounds from tlie soil. Thus it may be 
rendered [xjssible that only such small cpiantities remain dissolved in the 
cells as can exert a ben.cficial effect. 



Experinieiit with Rice in Soil Culture. 

In the following ex[)eri!uent with rice in soil culture the yellowing 
was not observed at all. The soil came from the experinient 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 i6g. 
sodium nitrate. Pot I received no further addition. Pot II v.as wateretl 
v.'ith 200 cc. of a o.i [)cr cent solution of ferrous sulphate, Pot III vvith this 
solution ami further with 2co cc. of a O. i percent solution of manganous 
sulfate. The seed was sown on May 24 and the number of shoots reduced 
four week's Liter to seven about etjually large ciies. The rice was cut 
on Nov. 10 with the followintr result : 



1 The observation ot .Lu> (I.e.) tliat a nuelcoinotcid containcil besides iron also maiigaiicsc may 
furnish us a clue in the right direction, /. 



i68 



Oi Loew and 8. Sawa : 





I. Control. 


11. Fe SO^ 


111. FeS()^ + MnSC\ 


Nunilier of stnlks 


19 


20 


18 


Average length 


5S.6 cm. 


59.7 cm. 


64.6 cm. 


Weight of straw 


45-7 g. 


46.5 g- 


48.7 g. 


Weight of grains 


5-7 g. 


7.0 g. 


1 1 .2 g. 



These numbers show that ferrous sulphate alone exerted a manuring 
effect, an observation also formerly made by others. Soils which contain 
iron ill a difficult soluble condition will respond in this manner. IMuch 
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 I and II. The stimulat- 
ing effect of the absorbed manganese was exhibited in an unmistakable 
manner. Nevertheless the iiifcrcnce 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 hckl 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 



» One of us (Z.) has mentional two years ago a case, in which tobacco plants showed no increase 
of the oxidizing power of the oxidases, after being irrigatal with o.i per niille Mn SO^ solution (Report 
No. 65 of the U. S. Dep't of Agriculture, p. 22). 



Oil llic Adioii of Miiiii^Aiieso ( *)III|mmiii<Is on i'laiit>. 



169 



highly diluted solution of manganous sulphate on March ir 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.ooi g. ferrous sulphate dissolved in ico cc. 
water was added. 

The plants commenced flowcrincj 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 ; 




Control plant?. 



Weight of the fresh fruits 
Wheight of air dry seeds 
Weight of the air dry straw 



60.5 

-3- 
, 10.7 



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 scant}' witli both the pea plants. 

Experiment -with Cabbage. 

A small plot of 12,5 squ. Meter received 3 g. manganous sulphate 
(anh)dr.) tlissohcd in 15 Liter water.' The lam! hail I'cccivcd the previous 
)'ear twice barn-jar-.! manure and had served for cultivation of barley and 
radish. This }'ear it received only ammonium sulfate at the rate of 50 g. 
to 12,5 square Meter. Cabbage seetl was sown on a part of this plot on 
April 25- Germination proceeded \cr\- slowly and many seeds failed to 
germinate. In the beginning of June, however, the diflerence in develop- 



* Two gram-; on .\piil 24 ; 1 g. on May :;i. 



lyo 0, Looiv aiiil 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 threatcnd, 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 \n many other cases it scenis almost impossible to find an 
explanation for the stimulant action ought not to deter us in every new 
case from searching for a clue. Now, it is well known tliat light retards 
growth. This hitherto unexplained phenomenon forms a strange contrast 
to the great chemical work tlie 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 tlie products of the sun's work are chiefly consumed. Tlie 
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 certain noxious 
compounds produced in the cells under the influence of light. Such 
compounds (Hemmungs-Stoffe, Ermiidungs-Stoffe) are frequently produc- 
ed in tlic course of the metabolism. ^ It is the office of the oxidizing 
cnzyms, as one of us (Z.) has suggested, to destroy noxious by-[)roducts 
of the benzene scries, a view verbally expressed as A^llows-: 

" The writer's view on this subject is that as the living protoplasm can 
oxidize carbohydrates and fat, but does not attack or attacks onl)' with 
difficidt}^ coiiipounds of the benzene group, and, on the other hand, as 



1 Cf. /')-. Reiuitzcr, licriclile cl. hotan. Gesellschaft, Vol. 11, p. 531 [1893]. 
^ Kciiort No. 59 of the U. S. Dep't. of Agriculture, Washington 1899. p. 27. 



Oil llic Aclioii (»r>I;i!i trill lose ( omiimhiikIs (mi I'iaiifs. 171 

just the opposite takes place with the oxidi/Jn<,r eri/.)-ins, it may be inferred 
that there exists between tlie 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 r,^enerally 
complete, but in this latter only partial." 

If the checking compounds are gradually changed by partial oxitlation, 
without being produced anew in darkness, we can understand the increased 
growth, in the absence of h'ght. Since, however, as we have seen above, 
the presence of manganese increases the oxidizing power of the oxidizing 
enzyms, the destruction of the ch.ecking 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 som.e 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 
{(hio) 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 
Iluppes biological law. The different behavior of fungi towards manganese 
in this regard seems to indicate that the enhancement of growth of 
pha;nogams 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 (A) would 
agree with this inference. 

The fust experiments on the influence of manganese salts (M\ lung! 
were made by //. Molisch.^ He concluded that manganese cannot 
enhance the development of flmgi like iron ?alts can. the latter even being 

* Wior.cr Akad. lior. (Vtolicr iS«)4. 



1 72 ^' Loew and S. Saiva : On the Action of Manganese C'oniponnds on Plants. 

indispensable. Recently Mr. TakaJiasJii 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 
oryza;. To this 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 oryz^. 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.i 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 oi MoliscJi, 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. 



■i 

i 



AT/./.. .I(;a'/c. CO/./., ro/.. r. 



/'/.<///■: x/j. 



-^ 







'l'al)lo showini; the iiillin-iKV ot iDaiit^auosi- on poa. I Mai)i,'anc>i.' \\.\\\\ 
11 Ctmtrol plant. To paj^i.- i6o. 



I 



Ueber die Wirkung des Urans auf Pflanzen. 



VON 

Oscar Loe-w. 



Tafel XIII. 



Die Lichtempfindlichkeit der Uransalze' liess es von Interesse erschei- 
iieii, die Wirkung derselben auf griine Pflaiuen zu verfolgen. da nioglich- 
crweise Spuren von Uran im Chlorophyllkorn die Umwandlung \'on Liclit in 
chcmische Energie befordern und dainit die chemisclie Leistung veriTiehren 
konnten. Es wurde von Seekamp beobachtet, dass Beriisteinsaure unter 
Vermittlung von Uransalzen durch Licht in Propionsiiure und Kohlensaure 
gespalten vvird. Eerner bilden die Fluorescenzerscheinungen und die in 
neuester Zeit beobachtete Radioactivitaet^ niancher Uran\erbindungen 
interessante Bezielumgen zuni Lichte. 

liis jetzt schcint nur cin einziger Versuch iibcr die Wirkung von Uran- 
verbindungcn auf I'flanzen ausgcfiihrt zu sein und zwar durch Kuop,^ 
welcher Uranylphosphat als Aufschwemmung in dcr Nahrloesung vcr- 
wendete. Knop zog aus dieseni Versuch C^zn Schluss, dass wegen der 
grossen Schwerloeslichkeit des Uranylphosphats dasselbe ohne jede 
Einwirkung sei. Sorgfiiltige Vergleiche mit Controlpflanzcn schcint cr 
nicht angcstellt zu haben. Zwar gehrjrt das Uranylphosphat mit zu den 
sch\verl()sh'chstcn Phosphaten, doch gcben Uransalze in eincr Verdiinn- 
ung von O.I pro niille keine EiUhing niehr mit IMonokahumphosphat. 
sondern nur cine schwache Opalescenz. In solchcr X^erdiinnung konnte 

* Bekanntlich wcnlcn l'rans;\l/.c aiich in ilcr riiotoi^inpliio vorwciulct. 

' Radioactive Substanzcn wirkcn im Duiikdn auf <.lic photoi^raphii-clio I'lattc <.mh uiul wcnn natli 
Monatcii diesc Ei;^cnschat"t crlischt, so kaiin sic durch IVlichlung luit Katluxlcnsti-ahleu wicvlcr bcr\or- 
i^crulcn wcrdcn. Die Kadioactivitacl sclicint lkvicliunj;cn 7U dcM l?o<iucrclsti-ahlcn und iwx Phospho- 
resccnz zu habcn. 

' Jahresbcr. 1". At^ricuUur- C'hcni. iSS.j, S. 130. 



174 



Oscar Loe>v: 



demnach wohl Uran audi in Gegenwart der Phosphate dcs Bodcns von 
den Wurzeln au%enoninicn wcrden. 

Ziuuicb.st wollte ich einigc Datcn bctrefTs des Giftigkeitsgrades^ 
samniein, dann die Wirkung bci sehr grosser Verdimnung beobachten. 
Auf jiinge Erbscnpflanxen wirlctc Uranyinitrat schon in 3 Tagen sclir 
giftig ein, aks diesc ju eine 0,2 proccntigc Locsung gesetzt warden. Wurde 
diesc Locsung bis auf 0.05%' Uranybiitrat vcrdiinnt, und junge Zwiebel- 
pllanzen eingesetzt, 5;o v/nr nach sieben Tagcn d,is Hanptblatt fast iiberall 
von der Spitze abwlirts bis nalie zur Hiilfte der Liingc gelb geworden und 
partiell \xrdorrt; die jiingeren Blatter wurden erst spiiter afilicirt." Die 
Wurzeln batten eine gelbliche Farbung angcnoniiiK'n, keiiie neuen Zweige, 
keine Wasserwurzeln waren erschicnen, wLihrend bei den Controlpflanzen 
in biosein Wasser dieses der Fall und i'tberliaupt das ganze Ansehen noch 
ein norniales v^ar. Ganz ahnlich war die Wirkung auf junge Gcrsten- 
pflanzen vo:i 12-18 cm. Hohe. Diese Pflanzen batten versucht, neue 
Wasserwurzeln zu treiben, diese waren aber schon als kurze Stummeln 
wieder abgestorben, wahrend die ControlgerstepHanzen in bloseni Wasser 
in sieben Tagen neue Wurzeln von bis zu 2 cm. Ll'inge getrieben batten. 

Wurden nun Erbsen- mid Gcrstenpflanzen in NiUirloesung gesetzt, 
welclier o.oi per millc Uranylnitrat zugesetzt war, so Hess sicli selbst nach 
Wochcn keine sch.adliche Wirkung melir v.-alirnelimen. 

Nun wurde (Febr. 16) ein Topfversuch mit Erbsenptlanzen ausgefiihrt, 
denen bis zur Beendigung der Bliitenperiode sechsmal je zwei Milligramme 
Uranylnitrat in 100 cc. W^isscr gelost, gegeben wurde. ^ Die jungen 
Pflanzen im Haupt- und Controltopf wurden auf fiinf moglich.st gicich 
grosse reducirt. Die l^liitenperiode daucste vom 2 1 April bis zum 12 I\Iai. 
Am 17 .Mai wurde eine l'liotogra[)hie aufgenommcn welche auf Tafel XIII 
re])roducirt ist und die iippigere Entwick'iung der UranpOanzen deuth'ch 



1 Auf Tliierc wirkcn bckaniitlich Uransalze sehr giftig ur.d rufca Diabetes, Degeneration der Leber 
und Paralyse hervor. 

2 Ein Vergleicli niit Manganuxyduisulfat zeigte, dass dieses weit >vcuigcr scluitilich wirlvte als das 
Uranylnitrat. 

» Dieser Versuch wurde gleiclizeilg mit deni M:inganversucli an Erlisen, und unter denselbcn 
Uedingungcn angestcllt (siehc don vorhergehenden Artikel). 



Ueber die Wirkiiiig <les Uraiis aiif I'flaii/cn. 



'75 



erkennen lasst. Auffallend war, dass die Uranpflanzen zur Zcit der Reife 
der Friichte neue Zweige aus dem Boden trieben, welche Bliiten bildeten, 
was bei kei'ner der andern zur selben Zeit beobachteten Pflanzen der Fa.\\ 
war.^ Am 2, Juni wurde geerntet, die Samen enthiilst und diese sowohl als 
das Stroh lufttrocken gewogen. Die Wurzelii batten in beiden Fallen nur 
wenige Knollchen entwickelt, doch die Controlpflanzcn immerhin etwas 
melir als die Uranpflanzen. 

Das Resultat der Wiigung war wie folgt : 





Fiinf Uranpflanzen. 


Fiinf Controlpflanzen. 


Samen 

Stroh 


29.5 g. 
'7,og. 


23.2 g. 
10,7 g. 



Ein stimulireiider Kinfluss des Urnnylnitrats niit \^crmclirung nicht nur 
des Strohs sondern auch der Samen ist demnacli 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 gpnz glcichen 
Verhiiltnissen ein Versuch mit Hafer angestellt. Ernto am 3, Jul:, Stroh 
und Samen (uncntluilst) wurden im lufttrocknem ZustauJe gewogen, mit 
folgendem Resultat : 





I r.mpriiiii/L'i). 


I oiitrolprtan/cu. 


/;ihl -In- Ihilmc 

Stroh,- 

Korncr mit I liiKcii 


1 1 

49-5 




45-^ 

-1 4 



Die forderntlc W'irkung des Urans ist SDiuit auch hicr unvcrkcnnbar. 



* Ausscr mil Mani,';uisull".U wareii rtlaiucn mit luvh vonluniUon I .>X'siiiii:on von K[ uml NaK 
bohaiiilclt wonlcii (sii-hc in iliosom Hoft Mr. .Iso's uinl Sinidrs Artikol). 



Ji 



Bur.r.. AGRii'. coTj.. vol . r. 



PLATE XIII. 




1 >\' I'.itl'l zois^t (Icn Kin(lu-< ikx I iTK. 1. H ^ I.-m crliiclt 0.012 i:. Ir.myl- 
M(r,\; 11. Controlpllanzo. /ur Scltc 174. 



I 



On the Physiological Influence of Manganees 
Compounds on Plants. 



BY 
K. Aso. 



With Plates XIV— XVII. 



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 nickeP 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. 5-5 cm. ingh. were placed, two 
shoots in each flask, in the following solutions : 

A. 0.02^ Mn SO^ + trace of Fe SO^ 

B. 0.02% Mn SO4 -f 0.020^ Fe SO, 

C. 0.020^ Fe SO 4 



Each flask contained further tl 
Ca(N03), 

KxNO, ... 
KHaPO, 

MgSO. .. 
(NH,), SO, 



■5 Cf. Oiii>, Jouin il it the Collide o 
•nr.il oth:i>-. 



e following nutrients : 

0.2 0^' 

0.15X 

o.o5>V 

0.05^ 

0.05^0 



-ciciKc, 111 p. Univ., Tokyo. \"o'. XII », jijirt L, a's> /?/i tr/iA- 



178 



K. Aso : 



In a second series, (a), (b), (c), the above solutions (A), (H) and (C) were 
diluted with ten times the volume of water, wliile the mineral nutrients 
were present in one fifth the quantity as in the first series. These shoots 
were kept in a cold room \\ith a winter temperature of 0^-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 : 





Xuml^cr of leaves. 


I.ength. 


Fresh weight. 






4 


1 1. 2 cm. 




A 




4 


lO.O „ 


r 1.2 grni. 


B 




4 


8.4 „ 


f 0-65 „ 




4 


7-2 „ 




C 




3 
3 


6.8 „ 
6.0 „ 


[ 0.35 „ 






4 


"•5 » 




a 






[ 1-3 >, 




4 


10.5 „ 




b 




4 
4 
3 


9-8 „ 
8.8 „ 
8.2 „ 


[ 0.9 „ 


c 




4 


8.3 „ 


[ 0.45 „ 



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 (5x7 m.m.) were well crushed in a mortar with addition of 



On the Physiological Influence of Manganese Couiponnds on riants. j-q 

lO c.c. of water. This extract (2 c.c in each case) served for the following 
tests :^ 

1. Upon addition of one drop of a 2% guaiac tincture, the blue color 
produced was more intense in tlie 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 i^^ 
h}'drogen 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 i 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-paraphen)Ien-diamine and hydrogen 
peroxid. 

Furthermore, two sections of equal size (4 x 6 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 exerts a 
more powerful oxidative power than the plants without manganese (C). 

Experiment -with Barley. 

On Nov. 26, barley shoots (7-S 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). 

* Ct". also the paj cr of the wiilor. "On oxidiViiiij Kiuyins in the \'egetahlc IVxly." in this bulletin. 



i8o 



K. Aso: 



A. 



Number of leaves. 



Length. 



19.5 em. 

11.6 „ 

17-2 ,, 

14.2 „ 

14-1 „ 

12.0 „ 

23-3 „ 

150 » 

17.0 ,, 

16.5 „ 

13-5 „ 

13-6 „ 



Fresh weight. 



0.70 gnu. 



0.65 „ 



0.55 „ 



The series (A), (B), (Cj, was no further observed, since some shoots 
commenced to show injury, and demonstrated beyond a doubt as also did 
the above described ex[)eriments of Loeiv and Sazva, that barley in water 
culture suffers — at least at the low winter temperature — from the concent- 
ration o{o.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. 


Fcbr. 2. 


I-Vl.r. 15. 


March 3. 


March 15. 


April 14. 




No. of 
leaves. 


Length. 


No. of 

stalks. 


length. 


No. of 

stalks. 


Length. 


No. of 
stalks. 


Length. 


No. of 
stalks. 


Length. 


No. of 
stalks. 


Length. 


a 

1) 
c 


8 

7 
8 


cm. 
27.0 

20.2 

1S.5 


4 
3 
2 


cm. 
27.1 

20.2 

22.0 


4 
3 

.S 


cm. 
30-5 

26.0 
234 


5 
5 
7 


cm. 
30.5 

32.0 
33-0 


5 
6 

7 


cm. 
31.0 

41.0 

390 


8 

7 
8 


cm. 
35-5 

52.5 

57-1 



1 



On the Pliysiological Iiifliieiice of ^laiiganesc Coiiipoinids on Plant*;. i s r 

The fresli weicrlit. 





Febr. 2. 


March 3. 


April 15. 


a 


4.0 grm. 


97 grm. 


16.8 grm. 


b 


1-5 M 


6-5 ,, 


16.7 ,, 


c 


2.1 „ 


S.5 „ 


17.0 ,, 



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 Febr. 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 
o.oi per mille Mn SO4 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 grent as in the 
case of the radish shoots. 



Experiment ^vith Wheat. 

At the same time at which the barley experiment was begun, one 
with wheat shoots, G-j cm. long, was started under the same conditions. 
The observations on the series (A), (B), (C), are contained in the following 
table : 



l82 



K. Aso : 





T\c. 20. 


Fc 


>r. 2. 




Numlier of 
stands. 


Lengtli. 


Fresh \\'eight. 


Number of 
stalks. 


Length. 


A 


( 3 


! 


230 cm. 
25.0 ., 


0.90 grin. 


8 


25.0 cm. 


B 


( 3 


-I 

( 


23.0 .. 
24.5 .. 


0.80 „ 


I 


17.0 „ 


C 


( 3 


i 


14.7 •' 

18.8 ,. 


0.50 „ 


I 


1 1.9 „ 



The leaves of this series gradually turned yellowi.sh, and since one of 
the two plants in h 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. 





I )lC. 20. 


Fehr. 2. 


Fei.r. 15. 




No. of 
stalks. 


Lx-ngth. 


Fresh 
^\eight. 


No. of 
staUvS. 


Length. 


Fresh 
weight. 


No. of 
stalks. 


Length. 


Fresh 
weight. 






cm. 


grm. 




cn^. 


grm. 




cm. 


grm. 


a 


( 4 


23-5 
24.5 


0.S5 


8 


2S.8 


6.6 


8 


33 -o 


9.1 


h 


1 ^^ 
( 3 


23.2 
25.0 


0.75 


6 


27.S 


6.0 


7 


30.S 


9.1 


c 


1 3 


22.1 
21.2 


0.75 


5 


23.0 


4.9 


6 


2S.5 


8.2 





March 3. 


March 15. 


March 29. 


Apr 


1 14. 




No. of 
stalks. 


Lengtli. 


Fresh 
^\"cight. 


No. of 
stalks. 


Ix-ngth. 


No. of 
stalks. 


Length. 


Fresh 
M-cight. 


No. of 
stalks. 


Length. 






cm. 


grm. 




cm. 




cm. 


grm. 




cm. 


a 


8 


38.5 


19-5 


8 


43-7 


8 


60.2 


34-5 


9 


67.0 


1. 


8 


39-0 


21.S 


9 


44-7 


10 


62.0 


46.5 


10 


72.7 


c 


9 


36.5 


19.0 


10 


45 -o 


10 


59.8 


40.5 


10 


62.S 



Oil tlie riiysiologioal Infliieiuc of Maiiieranpso ('oin]>oiin<U on l*li»iit>. 



183 



The ears d 


2ve 


oped in 


the 


followii^ 


S 


nunr 


iber : 








May 


I. 


May 


3- 




May 6. 


Ma)- 7 


a 




2 




6 






6 


7 


b 




3 




6 






6 


7 


c 



















I 



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 maiiy 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 MnS04 from March 3 to April 14 to 0.004^. 

Since now a parasitic fungus appeared on th.e leaves, tlie experiment 
was terminated on May 14. The final observations were as follows : 







Number 


Frcsli 


Total \\"eic;ht 


Weight 


\\'eight 


Weight 




Length. 


of 


\\-eight of 


of 


of 


of 


of 






car?. 


ears. 


dried straw. 


living leaves. 


dead leaves. 


dried roots. 




cm. 




grm. 


grm. 


grm. 


gi-ni. 


grm. 


a 


59.08 


7 


2.4 


5-6 


3-2 


2.4 


1.2 


1. 


59-09 


S 


3-5 


8.0 


7.0 


1.0 


3-0 


c 


4S.48 


4 


I.O 


74 


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 evidentl)- from the want of sufficient amount of iron, 
which was more evident in the later stage of development. It deserves to 
be pointed out cspeciall)-, that the wheat plant can overcome the injurious 
effect of manganese much more readily than the so closely related barley 
plant. 



1- The lirown points diil not here appear as distinctly as in the case with barley. 

■-' In comparing (A) w ith (I?) and (a) witli (b), it appears that the increase of iron had a couiitci*act- 
ing cftect upon the inilucnce of manganese, not only in reganl to the yellowing of the leaves but also in 
regard to the stimulating efiect, produced by the manganese salts. This same infeimce can be drawTi 
from the observations made on the railish sho<->ts (sec above p. 17S). The bivwn cv^Ior of the ixx>ts was 
due to some adhering MnC^j. 



I $4 



K. Aso : 



Experiment -with Pea. 

On March, 8, shoots of pea (3-4 cm. long) germin.ated 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 warm room. Of 
mineral salts, ferrous and manganous sulphates only were applied. 

a. 0.002% MnSO^ 

b. 0.002% Mn SO 4 +0.002% Fe SO 4 

c. 0.002% FeSO^ 

The following observations show the growth of the shoots : 









Length. 






P"resh ^\ eight. 
















March 22. 


March 27. 


March 31. 


April 7. 


April 20. 






cm. 


cm. 


cm. 


cm. 


cm. 


grm. 


a 


15.0 


iS.o 


23.0 


30.0 


39-2 


1.6 


1) 


11.6 


140 


16.5 


21.5 


30.5 


1.2 


€ 


1 1.6 


13.8 


17-5 


23.0 


33-0 


I.I 



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 tlio Physiological luflneiioft of Manganese Compounds on Plants. igr 



Conclusions. 

1. 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. 

2. Manganous sulphate added in a dilution of 0.002% to culture 
solutions 1 exerted a stimulant action upon radish, barley, wheat, 
and pea. Iron seems to counteract to a certain degree the action 
of manganese. 

3. The intensity of the color reactions of the oxidizing enzyms of the 
manganese plants exceeds that of the control-plants. 

» Tliis was of course transformed into phosphate in tlie cultui-e solution. 




« 






<^ 




A-rN- 



o 



-T 




o 



nUf.L. AGRR . ('('//. />'/. /■ 



PLATE .\7 7. 




'I.itr -bow in;; tin- inllui'iia- of in,>n'^.(in.Si' on wlicai. To l-.ii;i' iSj 



BULL. AGRIC. COLL. VOL.. I 



//.///•,. XI 7/. 



1 



/ 



'V 

-"(i 

L 






ria'.o show'ii!^ till' itilliiiMiiv of in;in;^,uH'so on |x\) -slioots. To [xis^'C iS 



On the Action of Sodium Fiuorid upon Plant Life. 



BY 



K. Aso. 



With Plates XVIII— 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. 

Ojw has in this regard observed that sodium fluorid in a dilution of 
0.000.03% li'^s a stimulating action on the development of algai.^ 



Experiment vrith Seeds. 

Seeds of rice, wheat and mustard (50 of each kind) were steeped in 
solutions of \%, 0.5%, 0.25%, 0.1% and 0.05% 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 : 



Xal\ 


Rice. 


Wlieat. 


Mu>t.ird. 


I % 


20 


I 





0.5 % 


28 


■» 





0.25% 


34 


■'y 





0.1 % 


38 


t 


6 


0.05% 


44 


7 


14 


Control. 


45 


16 


-3 



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 

* Journ. College of Science ; Imp, Univ., Tokyo. \ol. Xlll. \\\x\. I, (1900). With tungi, a stimu- 
lating action was obscrveil l^y him in the concentration of 0.005% X;^r. Veast is injuixxl by 0.01°^ NaF. 



1 88 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 off in length 
between these and the shoots from the seeds treated with the o.i^ and 
the stronger solutions. 

In a 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 % 

o.oi ,. 

0.005 ,, 

O.OOI ,, 

After 24 hours, the solutions were poured off and the seeds left to 
germination, 20 in each case. After 9 days, the following data were 
observed : 



Sodium fluorid. 


Number of germinated seed?. 


Average length of plants. 


Wheat. 


Soy-bean. 


\\hcat. 


Soy-bean. 


0.05 % 

O.OI „ 

0.005 » 

O.OOI „ 

Control „ 


15 
iS 

17 

19 
iS 


12 

20 
20 
20 
20 


plumule. radicle. 
7.ccm. 8.2cm. 
7.0 „ 9-6 .. 
S.5 „ II.O „ 

S.5 „ II.O „ 

7.0 „ 10.5 „ 


3.0cm. 

6.5 „ 
S.o „ 

6.5 ,. 

9.0 « 



Also in this case, therefore, like in the first, sodium fluorid in the con- 
centration of 0.05^ acted injuriously upon the seeds. 

Fxperiment with Soy-bean Shoots. 



Shoots of soy-been were (March 18) placed in solutions of sodium 
fluorid of the following concentrations : 



On tlio Action of Sodinni Flnorid npon Plant Life. 



189 





\al\ 


Fxngth of =lioot = . 


a 


0.1 % 


10 cm. 


b 


0.05 „ 


10 ., 


c 


O.OI „ 


10 ,. 


d 


0.005 ,. 


10 „ 


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 jellow. A slight touch sufficed to cause their 
dropping off. This indicates that the poison was retained to such an 
extent in the cot)'ledons that only traces reached the leaves above. 



Experiment ^vith Pea. 

On IMarcli 6, 1902, pea shoots 2 cin. long were placed in solutions of 
o.oi.o.coi and o.oooi^ sodium fluorid. The results are shown by the 
followinef.taSle : 







Lcni^tli. 




l-iv^l 


I \\ cii;l.l detcnuinol 










on A]iril 22. 




March 31. 


Ariil 14. 


April 22. 






O.OI % 


1 7.0 cm. 


20.4 cnl. 


20.4 cm. 




1.2 >-;:r.:. 


0.00 1 ., 


19.0 „ 


26.5 .. 


3-'5 •• 




1.3 ■« 


O.OOOI .. 


20S ,. 


30.3 -. 


36.5 .. 




I.^ .. 


Control. 


20.:: ,, 


20.0 .. 


;7.2 .. 




i.i .. 



A poisonous action on i:ea shoots is therefore produced by o.oi and 
0.001% NaF, but it is not any more noticeable when the dilution reaches 
0.0001%. 



190 



K. Aso: 



Experiments -with Barley Shoots. 

I. Slioots of barley were placed (March 25) in a culture solution to 
which was added NaF in the following proportions : 



a 


0.05 % 


b 


0.0 1 


c 


0.005 " 


d 


0.00 1 ,. 


e 


Control case 



The result is seen from the following table 





I.ength.1 


Number of stalks. 




March 25. 


April 4. 


April S. 


April 16. 


April 25. 


Apra 8. 


April 16. 


a 


29.3 cm. 


29.2 cm. 


28.2 cm. 


27.0 cm. 


4 


4 


4 


h 


37-5 •, 


37-2 „ 


37.5 >, 


40.0 „ 


4 


4 


4 


c 


30.0 „ 


31-0 » 


40.0 „ 


53-0 „ 


4 


4 


7 


<1 


28.2 „ 


30.2 „ 


33-5 '. 


43 5 ,' 


4 


7 


7 


e 


25.2 „ 


29-7 „ 


35-5 „ 


41-5 » 


3 


3 


3 



On March 30, the leaves of (a) lost their turgor while with (b) the 
tips of the leaves became yellowish. On April i, 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 ^^^'^ 
0.00 1 ^ was quite evident. 

II. On October 15, 1901, barley slioots 10 — 12 cm. long were placed 
in a culture solution to which were added : 



* The nieasurenient relates only to that part that still was ht-altliy. 



On the Vctioii of Sodiimi Fliiorid upon I'laiit Life. 



191 



0.0 1 



0/ 



^0 



Sodium fluorid. 



0.00 1 ,, 
0.000 ( , , 
Control. 



Tlic following table shows the observations made : 





I-cnt^lli. 


Nuinlicr 


of S 


alk^. 


l-a-h WViv;!.'. 




Oct. 31. 


Dec. 13. 


I'cbr. 6. 


Dec. 12. 


l-cbr. 6. 


Kcljr. r.. 


a 


15.3011. 


26.1 cm. 


27.0 cm. 


5 







1 1.5 !^''»". 


1) 


16.8 „ 


30.3 >, 


37-0 „ 


7 




10 


li.S .. 


c 


«9-5 -.^ 


390 :, 


— 


5 




— 


— 


d 


17-5 . 


341 - 


37.0 ,, 

1 


-^ 




7 


20.2 .. 



The i)lant in (c) whicli ticvelopcd most was injured b}' the sc\ere cold 
in the winter. 

Very many root-hairs and rootlets appeared in (c), also they developed 
very well in (b), but poorly in (<i). The remarkable effect of sodium tluorid 
in very high dilution u[)on an increase of tiie stalks was also here observed 
like in the former case with barley as the table shows. The accom})an)ing 
photograph (Tlatc XV 111) was taken on Dec. u. During this experiment, 
tlie solutions were renewed on Xtjv. 5» -5. and Dec. 17. 



Experiment •with Wheat Shoots. 



On May \S, shoots of wheat 6.5 — 7 cm. long, were placed (3 in each 
case) in soluticMis of o.ooi,*^ ^'>1'' (''^)i '"^'i^-' o.oooij^o NaF (b) to which the 
necessar\- mincial nutrients were added. Towards the end oi May, it 
became eviilent that the growth of the plants (a"* w.is much slower than 
that of (b) and of the control plants. Oa June u. the following data were 
t)bscrved : 



192 







K. 


Aso: 










Average number of leaves 
of one shoot. 




Avci 


•age length of 
shoot. 


one 


Total fresh weight 
of 3 shoots. 


a 


3 






2 1. 1 cir. 




0.8 grm. 


b 


4 






29.4 ,, 




1.6 ,, 


c 


4 






29.1 ,, 




1.6 „ 



A poisonous action of sodium fluoric! in tlie dilution of o.ooi^q ^^''^^ 
therefore quite evident, wliile in the 10 times hif^her dikition 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 — 10 cm. long were placed in solutions of 0.01%' 
and 0.0^% of NaF, but in the latter solution no development took place 
and the tips of the leaves turnetl yellow after a few days. In the o.oi/^ 
solution, some growth was observed, but less than in the control case, 
demonstrating the injurious inlluence at the dilution of o.Oi^^'o upon rice. 

II. On June 12, shoots of rice 22 — 25 cm. long were placed in 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 : 

a. 0.00 1 % NaF 

b. o.oooi ,, 

c. Control. 
A certain stimulant action is therefore noticeable in the case (a) and 

it a})i)e.\rs 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 NaF of various concentration. The following 
data show the result ; 



I'otal numl^er 
new rootlets 


of 


Total number 
of leaves. 


1 .ength. 


32 




10 


32.0 cm 


37 




8 


30.4 .. 


20 




6 


33-0 ,, 



I 



(hi 1li<> Action of Sixliiiiii Fhiorid upon IMiiiif Fiil'o. jq? 

XunilxT of flower? ilcvclopci. 

y ^ 

T'cl)r. 25. Fcbr. 27. 

O O 







Fcbr. 24 


a. 


o.oi % Sodiuin fliiorld. 





b. 


o.ooi ,, 


^ 




c. 


O.OOOI ,, 


2 


d. 


Control. ,, 






4 6 

3 3 

^ 3 

There liad no bud opened in the sohition of 0.01%' NaF, which shows 
a poisonous influen.ce 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 u-e 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 eftect 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 lla//, 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. Tiie result after 
37 da}'s was as follows : 





Orii^in.nl 
number of 1 


)U<ls. 




N 


mnbcr of 1 
openeil. 


U(l> 


Roin.irk? 






>ri^i71 


7" 


Marcb 27. 


.\pn"l 10. 




a. 


O.O! _Pq Sodiiu-n fluorid 5 












I 


lx.-ives 
Ycnnw. 


b. 


O.OOI ,, ,, 4 




4 




4 


4 




c. 


O.OOOI ,, ,, 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). 

Mere also a ceitain accelerating influence of sotlium fluorid in very 
high dilution upon the de\'clo]">ment c->f leaf-buds was evident.' 

* It may be im-ntioiKNl licro tint also some pxiH.M-inioiit# were ni.iile with iiijcctioiis of a 0.2^0 
solution of soilimn lluorM into yoiini; branches anil but!*, but bo>iiUv the ilcath of the tissue .nrouixl 
the point of injection no strikinsj efkvt w.-\s noticed. 



3 94 



K. Aso: 



Experiment with Pea in Soil Culture. 

Two pots each lioldinj^ 2.3 k. soil served here for the experiments 
with peas, of which 15 seeds were sown on Febr. 19. The j-ounj^ plants 
were reduced liowever, on March 27, to five equally larc^e ones in each 
case. Each pot had received the followin^^ manures : 



4.6 grin. 
3.0 ,, 



Common superphospliate. 
Potassium carbonate. 
Sodium nitrate. 



While the plants in one pot were treated with sodium fluorid, the other 
pot served for control. The pots v^ere kept in the green-house almost all 
the time. The ainount 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 it, 25, April 14, 21, 2S and May 6. Tlie total amount 
of sodium fluorid was therefore 0.006 grm. and still in spite of <"his 
small quantity, a stimulating effect was gradually noticed and was finalh' 
also recognized by the weight of the seeds produced. The formation of 
flowers commenced on April 22, and was ended on the i6th of May, One 
day afterwards a photograph was taken (see Plate 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 evcr>' da}' 300 c.c. water for 
irrigation was applied, later on the quantity was increased to 500 c.c. 
The fruits had ri[:)encd on the 2nd of June, and were weighed in the fresh 
state, while the straw was weighed in the air dry state. Tiie results wcrj 
as follows : 



1 ( )ii Ai.ril 23, sonie plant-lice niado tlic nppe.araiicc on the leaves and from now careful search 
was kept up ;incl every louse noticed Killed ]>)• touchinjr tlie insects witli a little brush moistened with 
a 1% carlioHc acid solution. 



I 



On llii' Ac1i(»ii <>r Sixiiiiiii Fiiiorid ii|)oii IMaiil MIV 



195 




•"ontrol. 



Weii^lil of iVcsli fruits. ... 
Weight of air dry seeds 
Weight of straw 



-J- 

10. 



This result doiibtlcs.s .sho^v.s that .1 stimulatiiifr action b\' thi.^ small 
quantity of fluorid had taken jolacc. 



BULL. AGRIC. COLL. VOL. V. 



rLATL. XIX. 




T. 11. 

Plate slunviii;.^ tlio iiilliR-iicc ol" s idiiiiu liiiorul o-i |xm. 1 Fluorine j>Iant 
II control phuu. To jxis^o lo^. 



k 



On the Action of Sodium Silico-fluorid upon Plants. 



IJY 



K. Aso. 



The forctjoing observation with sodium fluoiid niiidc it very jirobablc, 
that sodiuni sih'co-fiuorid 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 % vSodiuni silico-fluorid. 

b. o.oi ,, ,, ,, 

C. 0.005 ,, M ,, 

d. o.ooi ,, ,, „ 

c. Control 



The observations were as follows : 





Soy-bcaii. 


r.avlcy. 


March 27. 


Shoots (a) anil (1)) lost turs:;or. 




„ jS. 


Shoots (c) lost turgor. 




,. jO- 


Shoots (d) stationary. Con.Milcrablo 
(Icvdoimicnt in the control case. 


The tips h;ul all turncxl yellow with the shixits (It) 
and (c), wliile with (a), all leaves had withcrcil. 
I.eaves with (d) stil! normal. 


April 1. 


Shoot (il) lost tmgor and w iilicrcd. 


Almost all leaves with (1)) anil (c) aiv dead ; only 
shoot (d) and the amtrol sho«.it were still healthy. " 


„ 8. 


Only control shoot still alive. 


Control sluK>t normal, has thrown 10 cm, sIkvM (d^ 

still alive, Init stationary. 



if,8 K. Aso: Oil the Attiou of Sodiiiiu Silico-UuoriU upon Plants. 

It will be noticed that sodium silico-fluorid is a stronger poison than 
sodium fluorid when tlie 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. But a remarkable effect was noticed with barley shoots (d), i.e. 
where the dilution of sodium silico-fluorid was O.OOi,^^. 

While here growth in hight \\as vcr\' sluggish there had develo])ed 
up to the 8th of April from the originally th.ree stalks as many as seven 
new stalks, while not a single new one had started in the control case ! 
This fornis a third instance of tliis kind of action of a fluorine compound, 
since sodium fluorid produced tlie same plienomenon with barley shoots in 
the two cases described in the foregoing article. 

It deserves notice, further, that soy-bean is more easily injured by 
higidy diluted sodiuni fluorid and silicc-fluorid than barley. 



On the Action of Highly Diluted Potassium iodic! on 
Agricultural Plants. 

BY 
S. Suzuki. 



With Plate XX. 



Since it became known that in certain giands, espcciaUy in tlie 
thyroidea, occur pecuh'ar proteins containing iodine, the inference had to 
be drawn tliat the vegetable food for animals and man contains a small 
.imount of iodine compounds. The analyses of the ashes of crops thus far 
made ignore the presence of iodine completely, but a few instances are 
known of iodine having been found in other pha^nogams than agricultural 
l)lants. Boitrcct^ found iodine in ashes of plants growing on soil 
containing for lOO kgr. 0.83 mg. iodine. LiliacciC and Chenopodiaceie 
took up comparatively more iodine from this soil than SolauactW and 
Uinbclli/c-rce. Long ago, however, it was known that marine algai contain 
iodine.'' Stanford^ observed in the dr\- matter of such an alga, Laminaria 
digitota, 0.453^ iodine in average, in that of Fiicus scriatus 0.085^0 ^'""^ 
\u \.\\\xi oi Fiiciis I'^siciilosiis o.02g%. In which form the iodine is present 
in these plants is not \ct fully decided, but it is very probable that 
it forms a constituent of peculiar proteins. If this amount of iodine 
would be present \u form of <i soluble iodid it would probably act poison- 
ously, at least, if the cell sap would be of an acid reaction. Algx with a 
neutral cell sap, as S/^irogyra, show considerable resistance to sodium 
iodid. It has first been observed by Pirl's^ that potassium iodid even xn 

* L'ompt. iviul. Vol. 129 [1S99]. Also llot. CentralM. 1900. No. 2;. 

* The ashes of these alijiv scrvcti long since for the nianufactuiv of iodine. 

* Quolo.1 in MusjTiill's Tccluiical ChcnuVtiy. 

* I. 11. t". Av;ric. Chcniio. iS6S \>. 2S9. 



200 S« Suzuki : 

a dilution of 0.0625 per mille proved very injurious for pha^nogams. 
Young maize plants died after two weeks and soon afterwards also buck- 
wheat plants. Cress resisted longer but did not produce any ripe seeds,' 
LoeiV' has also observed a highly poisonous action of sodium iodid on 
buckwheat seedlings. Recently also A. Voelckey"^ made some experiments 
in this direction. Sodium iodid at the rate of 200 cwts. per acre killed 
wheat, barley and red clover, peas were slightly benefited. At the rate 
of I cwt. per acre it injured wheat and barley. A top dressing at the rate 
of 4- cwt. per acre injured also wheat and barley. Soaking the seed in a i per 
cent sodium iodid solution increased however the yield of wheat and barley, 
grain and straw, and benefited the red clover. In water culture sodium 
iodid proved highly poisonous ; thus even in a dilution of i : 43/00 it 
caused the roots to be " quite dwarfed." Since the general occurrence of 
iodine in agricultural products must be assumed and consequently also the 
occurrence of iodine traces in every soil, it seemed to me of some interest 
to observe the effect of a small increase of traces originally occuring in the 
soil, since poisonous compounds can in very high dilution often produce a 
stimulating action {H'uppes biological law). 

I have tried, therefore, the culture of pea (Pisum) in the soil of our 
College farm under the influence of small doses of potassium iodid. ILnch 
pot contained 2300 gr. air dry soil and was inanurcd with 3 gr. NaNO.,, 
3 gr. Ko^Og and 4.6 gr. common superphosphate. Fifteen seeds were 
sown on Feb. 21st and after the young shoots had readied about 15 cm., 
they were reduced to 5 equally large ones. 

While one pot served as control, the main pot received o.ooi g. 
potassium iodid on each of the following dates : ]\Iarch 1 1, and 25, April 
14, 21, 28, and May 6. The total amount of potassium iodid was therefore 

' This interesting diflcrcncc hclwccn the cress on tlic one liaml and huckwheat and maize on llie 
other iiiighl 1)C explained l)y the assumption that tlie iodine, when liberated from the compound, is 
ahsorljed hy the allyl coni])ounds ]n-oducetl in ilie cross l)erorc it can seriously injure the dying 
jirotoplasm. 

- lie found thai in a culture solution containing 0.2 per mille sodium iodid, young buckwheat 
])lants could not grow at all (Ein naliirliches System der Giftwirkungcn p. 108). 

^ J. Roy. Agr. Soc. Engl. [Ill], 11, 566-591., Abstract in the Journ. Clieni. Soc. May, 1901. 



Oil llio Action ol" Ili^Hily I)ilii1('<l l'oljiN>iiini lodid on A^'iitiilt nral IMiUiN. oQl 

only 0.006 gr. Still, this exerted a stimulant action as the further develop- 
ment revealed. The formations of flowers commenced on A[/ril 22, and 
had ended on the i6th of Ma)'.' One day afterwards a photograph was 
taken (see Plate XX), which shows that under the influence of potassium 
iodid the plant reached a greater height than the control plant. Up to 
the flowering period the pots received almost every day 300 c.c. water, 
later on 500 c.c. The fruits had ripened on tlie 2nd of June, and were 
weighed in the fresh state, while the straw was weighed in the air dr\- 
state. 

The results were as follows : 

Potassium iodid. Coinro!. 

Weight of the fresh .fruits 72.4 gr. 60.5 gr. 

Weight of air dry seeds 26.3 ,, 23.2 ,, 

Weight of air dr)' straw 15.5 ,, 10.7 .. 

This result doubtless shows that a stimulating action of this small quantit\- 
of iodid had taken i)lace. 

^ At that time aphides made Iheir appearance on the leaves, they were easily killeil i)y touchini; 
them with a fine hair l^ush moistened with a carholic acid solution of i%. 



J 



llifj.. .ICAWC. cor I.. VOL. ! 



rr.iTE XX. 




T. II. 

Plato si owino; the innucnco of irtiui of sodium on jxm. 1. lolinc plant 
II. Control plant. To paij;o 201. 



i 



On the Poisonous Action of Potassium Ferrocyanid on Plants. 



RV 



S. Susuki. 



The observation of Knop that chlorotic plants turn green not only by 
ferric salts but also by potassium fcrroc}'anicl, led me to try whether the 
iron in this last named form could not be used with advantage in water 
cultures, since the iron would retain its solubility in presence of phosphates, 
whereby its absorption would be facilitated. Kjiop'^ observed, however, 
not onl)'' a useful but also an injurious action consisting in the stoppage o^ 
growth when he applied the potassium ferrocyanid in a dilution of O.i per 
mille.f 

Since, however, it seemed probable that this injurious action might 
be avoided by a higher dilution of that compound, I have applied a 
dilution of o.oi per mille. The solution had the following composition : — 

Calcium nitrate (anhydrous) 3. per mille. 

Potassium nitrate i 

Magnesium sulphate (cryst.) 1.5 ,, 

!\Ionopotassium phosphate. i. ,, 

This solution received in the control case (a) 0.02 per mille ferric 
phosphate in fine suspension while in two other cases, (b) and (c), o.oi per 
mille potassium fcrroc3'anid. Tlie culture liquitl (c) c<~>ntaiiied further o.f) 
l)cr mille ammonium sulphate. 

Young barle)' shoots were placed in those solutions on Dec. 14th, 
and kept in a room near the window, at a temperature ranging from 4° 
to I5^C. After a few weeks a very marked difference was noticctl. The 



* J. H. 1. Ai^r. L'l><.in. iSoo \\ -(^'^- -^•-"<^. ''>'^1. 1SS4, p. 140. 

•j" Algcc are not injiirol hy this ililutioii. but hy 0110 ol 0.5^,^ (('. ^>.7«', System ol \H>isonous 
actions p. 55.) 



204 i^' Suzuki: 

roots of those plants wliich had received potassium ferrocyanid showed 
much less development than the roots of the control plants — they liad fewer 
lateral roots and less root hairs. Gradually also a yellowin<^ of the leaves 
set in. leading to death. On Feb. 3rd the still living leaves were counted, 
the length of the longest leaves measured and further the total weight of 
the still living parts of the leaves and the roots determined. The results 
were as follows : — 





Xumlier of the living 
leaves. 


Length 
longest 


of the 
leaves. 


Total fresh wc 


a. 


8 


21 C 


m. 


4.4 grm 


a. 


10 


21 




3-6 ,. 


b. 


6 


18.5 




1.8 „ 


b. 


5 


17.0 




1.9 .. 


c. 


0^* 


12.0 




1.5 .. 


c. 


Q-:f-» 


13.0 




1.0 ,, 



It seems therefore that the ammoniuir. ferrocyanid probably formed in 
the solution (c) is still more poisonous than the potassium ferrocyanid. M\^ 
result shows that the ferrocyanid compounds are exceedingly injurious even 
in the high dilution of O.oi per mille. I also have noticed like Knop the 
formation of some prussian blue on the surface of the roots. It may be 
asked, in what this poisonous action of potassium ferrocyanid consists. The 
supposition suggests itself that this salt is split in the plant with the produc- 
tion of hydrocyanic acid and that this is really the poisonous principle. i' 
This is rendered still more probable by the observation of Knop that 
the potassium ferrocyanid as such is changed very soon in the juices of the 
plant. But on the other hand there exist cases in which hydrocyanic acid 
appears to act less poisonously. Some plants in Java produce free lu'dro- 
cyanic acid in the course of a metabolism peculiar of their own. Thus the 
leaves of Pangiuvi cdnle are said to contain as much as \% of hydrocyanic 
acid for the dry matter. 

* and ** : All leaves were dying. 

f Z(?^7i' and Tsiikamoto(^^\\\^ I'ulletiii, \'i>i. II, No. i) ohservcd a i.oisonous action on germinating 
seeds of hydrocyanic acid in a dilution of 0.2 per niille. A solution of i per mille l<ilk'd young radish 
plants in 15 hours. 



On tlu' roisoiioiis Action of i'otii.ssiiiia IVrrocyaiiiU on I'laiits. 20 ' 

Furtlicr there exist several glucosides in plants, such as amygdalin in 
the bitter nlinonus, in Pygimn parvijloruin and Gymueina latifoliuin, and 
linamarin in the hairs and seedlings of Liiiuin nsilatissimmn which gliico- 
sides yield on decomposition by enzyms also hydrocyanic acid. Soave 
has shown that bitter almonds produce h)'drocyanic acid from amygdalin 
during germination.^ Liitrj- proved the formation of hydrocyanic acid 
in the seedling of a Japanese Visciim. It also has been found \\\ the roots 
of ManiJiot ntilissiina and Vicia and in still other plants, as Passiflora 
qitadra)igulata and Colocasia gigantca. 

An exposure to very dilute gaseous hydroc}'anic acid for a short time 
will not injure plants, but it will suffice to kill noxious insects investing 
the plants A. F. Woods and others recommend therefore the treatment 
of invested 2)lants in greenhouses with hydrocyanic acid gas. 

As the main results of my tests follows : — Potassium ferrocyanid is — 
even in high dilution — not suited as a source of iron for the chlorophyll- 
bearing plants, since it will gradually injure the plants and even the 
chlorophyll itself. 

* It may serve there as a protection against depredations by insect-;. If it is decomposed in formic 
acid and annnonia in the seedling, the latter may again ]jc utiiizctl for building up iirotcins. 

* IJuIl. See l)ot. lie IVance, 181)7. 



On Oxidizing Enzyms in the Vegetable Body. 



liY 



K. Aso. 



Introductory Remarks. 

Although various iiu'estigations on the oxidizing actions b}- enzyms 
liave been pubh'shcd during the past ten years, certain questions are still 
unsolved, especially in regard to the identity of the enzyms which cause 
various color reactions. The following observations in regard to this point 
may therefore be not be without some value. 

The fact that the juices of many fresh vegetable and animal objects 
can produce a blue color with guaiac tincture on addition of hydrogen per- 
oxid was known long ago but the true cause was recognized but recently. 
YosJiida^ was the first author wIuj pointed out that an especial cnz\'ni with 
oxidizing actions is contained in the sap of the lac tree,- to which is due 
the blackening of this sap in contact with air b\- the o.xidation of urushic 
acid, the most important constituent of the sap, to oxy-urushic acid. 
Bcrlraiid'^ wiio contiiuicil the study of that sap called this cnzym, laccasc. 
He aiul l\utrqitclot demonstrated lurther the witle ilistributi(->n of this 
laccase in the vegetable kingdom and tl.at it can [iroducc a blue reaction 
with guaiac even in absence kA h\clrogen peroxid. Ray-Patlhadc^ ascer- 
tained the presence kA laccase in germinating seeds. While laccasc iloes 
not act on tyrt>sin, a s[)ecial en/ym called b\' /)\7V;<f/.'</ tyrosinase, '" changes 

» Voshidij. Jouni. CIkui. Soc. Xl.lll. 1SS3. 
^ This juice is called ' Urushi ' in J.ip.in. 
^ Hull. S\x". chilli. II. 1'. 717, 1804. 
* Compt. rciul. 121, \\ iiOJ, 1S95. 
" Hull. Soc. I'him. 13, p. 793, iS<)(). 



208 



K. Aso: 



tyrosin to a red and ultimately to a black substance. This enzym is 
present in the root of Dahlia, the tubers of potato and some fungi, as 
Russula. 

Martitiaiid^ observed a laccase-like eiizyni, cenoxydase, in grapes 
and other fruits, and ascribed the cause of discoloration of red wines to 
this enzym. Recently Toloniei^ showed that a similar oxidase is present 
in several yeasts and has close relation to the production of the bouquet 
of certain wines. Brcajtciaf^ found an oxidase in the leaves oi Isatis, that 
causes the oxidation of indican to indigo ; Toloniei,^ an oxidase, called 
olease, in ripe olives that causes an oxidation of the olive oil ; SartJiou^ 
observed an oxidizing enzym called by him, shinoxidase, in the latex of 
Schinus inoUc. Dubois^ ascribed the cause of the production of light by 
certain animals and plants to an oxidizing enzym, which he called 
luciferase. Lepinois studied especially the occurence of the oxidizing 
enzym, which gives a blue reaction with guaiac and hydrogen peroxid and 
called \\. peroxidase. Recently, Raciborski'^ studied the distribution of the 
same enzym in certain plants. The name leptomin given by him to this 
enzym is however not justified, since Lepinois had named it already 
peroxidase. Similar investigations were made by Griiss. Loeiv^ who 
observed this enzym and laccase in the leaves of tobacco, attributes the 
changes of these in the curing and fermentation process to their actions.^ 
Woods'" ^^ observed an increase of the oxidizing enzyms in plants patholo- 
gically affected. U. Snziiki^^ investigated the relation of oxidizing enzyms 



1 CA)m])t. Rend. 124, p. 512, 1897. 

* Real. Acad. Line. 1896, 5, i, y. 52. 

3 Conip. Rend. 127, p. 769, 1898. 

■• Real. Acad. Line. 1896. 

" Journ. riiaim. Chim. 6 ser. 12, p. 104 and 11, p. 482, 1900. 

« C. R. 123. p. 653. 1896. (Carl Oppcnheinicr : Die Ferniente u. ihrc Wirkungcn. p. 301). 

■> 15er. d. d. I3otan. Ges, XVI. 119. 1898. 

" Repfiit. Xo. 65. U. S. Department of Agric. 1900. p. iS. 

" Tiiis socalled fermentation is not due to bacterial action as Locn' has abundantly proved. 

in C. Bakt, 11. Abt, 1S99, p. 745. 

n 15ull. College of Agric. Tokyo. Vol. IV. Xo. i. 



On Oxidizing Enzyins in tli<' Veerctiiblo Body. 20Q 

in the case of the mulberry dwarf disc.ise. Lately, Hunger'^ observed the 
presence of oxidase and peroxidase in the milk of the cocoa nut, Nezi'ton 
and tlic writer in the tea leaf. Recently a particular enz)-m, called 
spermase, was found to exist in resting barley- and yeast^ by Gruss. A 
new enzym, catalase, which is of general occurence was discovered and 
fully studied by Loeii'.^ 

As to the physiological role of Oxydase (laccase) and peroxidase, 
Loetv has pointed out that it very probably consists in changing by partial 
oxidation injurious by-products of the benzene series, generated in the 
course of metabolism. Catalase further can perform the highly important 
function of destroying h}-drogen peroxid which may be produced as a 
by-product in the course of cellular respiration. Many investigations have 
indeed shown that, whenever the molecular o.xygen of the air serves for 
oxidations at the common temperature, hydrogen peroxid is produced as a 
by-product. 

The Tests for Oxidizing Enzyins Applied in 
this Investigation. 

Although there arc known quite a number of color tests upon oxid- 
izing enz\'ms, onl}- the following served here for comparison. 

1. Test icitli gitaiac tincture. 

According to Bcrtrand,^ an alcoholic solution of guaiacum resin turns 
blue even \\\ absence of hydrogen peroxid, when a trace of laccase acts on 
it in presence of air. When the laccase is present in a larger quantity', the 
blue color will turn to green and then to jellow. It is well known that 
this blue reaction ma\' be obtained also with \arious o.xidizing agents, 
such as lorric chlorid, chlorine, nitrons acid, pctas^^ium terrocyanii! .uid 

^ Hull. do. riiistitul l{ot.»nii|Ut' ill' Uuiti'ii/ori;; No. \'lll. 

' Wtxiieusclir. 1. Hiauiiri. iSgq, No. 40; Hot. I iiitr. lUt. i.xji. No. i. p. «i, 

* Hrewer's Jouin.il. \'oI. XW. No. 10. 11. \\^. looi. 

* Report. No. OS. V . S. 1 Vp;iitniont of .■\giic. 1901. 

* Coinpt. Rcml. 120. i>. 166, (1S95). 



2IO 



K. Aso: 



some salts of the heavy metals, also with quinoiic. ScJidubein also mention- 
ed that pure mercury^ and the noble metals produce this blue reaction. 
Traces of alkali are not favorable for the development of the blue guaiac 
color by laccase ; neutral or slightly acid solutions are best suited. 
Certain organic substances will also interfere with this reaction, as 
mentioned farther below. 

For my tests I applied a frequently renewed guaiac tincture of 2%, 
kept in small flasks of colored glass, protected against direct sunlight. 

2. Test zvitJi guaiac tincture and hydrogen per ox id. 

The !>lue reaction which is obtained with guaiac tincture and h}'drogen 
peroxid was observed long ago by Theuard, and Blanche and Taddei. 
Schonbein ascribed this reaction to all kinds of enzyms and for a long time 
it was also applied as a test for the common diastase. It was but recently 
that Schonbein' s view was found iiiCorrcct by Bertrand. Schonbein further 
had entertained the view tliat the power of fresh animal and vegetable 
juices of decomposing h}drogen peroxid witii development of oxygen, was 
a property of all cnz\'ms, but also this conclusion was recently proved to 
be erroneous. 

Spitzcr- ascribed the decomposition of hydrogen peroxid to the same 
enzym which produces a blue color with guaiac tincture and hydrogen 
peroxid, what did not agree with Lcpinois^ observation that there is no 
proportional relation between the intensity of oxygen development and the 
intensity of color reactii-ns such as with guaiac tincture, guaiacol etc. 
I'^inall)', Loc7v' jirovcd that the property of catalysing hydrogen peroxid is 
due to a sj^ecial cnzym, called by him, catalase,'^ which frequently is 
present as an impurity in other enzym preparations. In order to test for 
peroxidase, I prepared a solution of hydrogen peroxid by. dissolving 



1 I liave obcrved (hat tliis l)luc coloration did not increase upon addition of hydrogen peroxid, 
hence its jjroduction can not be due to a suiiposed formation of hydrogen peroxid liy mercury. 
- rfliiger's Archiv. Vol. 67. 1897. 
3 Compt. Rend. May 26. 1899. 
* Report of U. S. Department of Agriculture. No. 68. 1901. 



On ()\idi/iii!^ Eiizvins in flio Ve)?etabl«' Body. 211 

5 grms. of sodium peroxicl in 200 c.c. of water containing a little more than 
the calculated amount of sulphuric acid, in order to provide for a faint acid 
reaction, indispensable for the preservation of the hydrogen pcroxid. 
Tincture of guaiac was prepared by dissolving 2 grms. of good transparent 
guaiacum resin in lOO c.c. of absolute alcohol. 

In testing for peroxidase with guaiac, tiie solution must be always 
neutral or of a slight acid reaction as already pointed out. Special 
precaution must be paid to have the solution of guaiac frequently renewed, 
since an old one will cause a blue reaction with hydrogen peroxid alone. 
Since much hydrogen peroxid will injure the oxidizing enzyms, care has 
to be taken to apply only very little. 

3. Test zvitJi glial acol and hydrogen peroxid. 

The color reaction with guaiacol was first applied b)' Boiirqiielot.^ 
Recently Diipoiiy- observed an oxidase in the sali\'a which produces a red 
color with guaiacol and hydrogen peroxid. Like him, I also made use 
of a \% aqueous solution of guaiacol. ^ In cases of mere traces of the 
corresponding enzym, a red coloration appears after 5-10 minutes ; other- 
wise, at once. 

4. Test with Pa ra ph e nv leii dia )n i >ic and hydrogen peroxid. 

A color reaction of vegetable objects with paraph.enslendiamine hat! 
first been observed by Bonr(]in'lot.'^ That base was applied b}' Storch^ 
for a test upon fresh milk w hich yields a deep violet color (mi addition o( a 
salt of that base and h}'drogen pero.xid, while boiled milk fails to give this 
reaction. The writer tested the beha\ior of this reagent with many 



1 C. R. Hoc. biol. 46. p. S96. 1890. 
-' Jahrcsbcricht f. Agric. Chem. 1890, II. [-. 440. 

3 This si)lutie>ii must be slightly acid. < »tht r oxidi/iiig cn/yius as well as certain comjx^uiids 
(kiesols) require a weak alkaline medium. Schinoxidase acts best in a neutral mcxiium (S.irthou). 
* C. R. Soc. biol. 46. p. 496. 1S96. 
<* Jahresber. f. Thicr-Chem. 28, p. 256. 



212 K. Aso: 

vegetable objects and found tliat, when tlie reaction is successful, a green 
color is first produced, which turns to dark violet ; in only a few cases it 
was other .vise. In my tests I have always apph'ed a 2% aqueous solution 
of hydrochlorid of paraphenylendiamine freshly mixed with an equal 
volume of a 0,8^ of sodium acetate solution. The acetate gives the 
reaction better than the hydrochlorid (In absence of an oxidizing enzym 
the acetate produces itself but \'ery slowly a reddish brown color. Control 
tests are required). 



5. Tesi witJi TetyaitietJiylparaphenyleiidianiine. 

Griiss^ applied at first paper which had been soaked in an alcoholic 
solution of tetramethylparaphenylendiamine. In his recent work" on 
yeast, the reagent was employed in the following manner : ' a granule 
of tetramethylparaphenylendiamine of the size of a pin head was dissolved 
in four c.c. of water. The solution was allowed to flow on to a piece of 
filter paper, 5x5 cm., so that the paper, which can be most conveniently 
placed in a Petri dish, is thoroughly saturated,' immediately before the 
objects were tested.^ In my case, the chlorid of tetramethylpara- 
phenylendiamine was used instead of the free base, and the tests were 
carried on according to Gruss's method.'* I observed this color reaction 
with several vegetable juices.^ Upon addition of a few drops of a o.\% 
solution of the said salt to an extract of malt, or the root of radish, a 
violet coloration set in immediately, but not in the control case. 



1 Journ. (hem. Soc. 1901. i>. 33. 

2 brewer's Jouni. Vol. XXV. 1901. Me tailed this paper tetra paper, ami the [taper iDoisteiied 
with solution of the chlorid of tetrainethylparaphenylendiamine and soda, tetra soda paper. 

3 He wrote in his latter article, that chlorid of tetramethylparaphenylendiamine gave an exceed- 
ingly strong reaction in the case of resting harley. 

■* I <litl not use tetra soda paper. 

Tetramethylparaphenylendiamine is easily colored vink-t liy the air alone, so that a control test 
has to be carried on in every instance. 



I 



Oh 0\i«liziiit? Enzyme in llio V«j?etal>le IJody. 213 

6. Test 7vith tetrainet]iylpay(Xpheuyleudinniinc and 
hydrogen peroxid. 

This test had not been in use previously. During my search however, 
for spermase in seeds and other vegetable objects, I noticed that a section 
of potato failed to give a reaction with an alcoholic solution of tctramethyl- 
paraphenylendianiinc alone, but a beautiful violet color upon addition of a 
drop of peroxid. This phenonienon led nie to repeat this test with milk; 
and indeed I observed that fresh milk behaves exactly like the potato in 
this regard. This color sets in at once while in absence of the enzym the 
coloration appears but very slowly. 

Boiled milk fails to give this test. Hence, I propose this reaction as a 
useful test for distinguishing fresh milk from boiled milk^ 

Behavior of Various Objects. 

The first series of tests were made with slices of \egetable objects and 
the second with the juices of plants. 



* There is the so-called iiidopheiiol reaction fi>r axidases consisting in the production of a blue 
color with a mixture of a-naphtol with paraphenylendiamine and sothuni tarhonatc. This reaction is 
however not very delicate and appears slowly also in absence of oxidases. 



214 



K. Aso: 



Slice of 


a. 

Guaiac 
tincture 

alone. 


b. 

Cluaiac 

tir.cturc 

+ ii,o,. 


c. 
Guaiacol 
+ II2O.,. 


<1. 

I'araphcny- 
lendianiine 

+ IL(\,. 


e. 

Teti-amethyl- 

paraphenyl- 

cndiaminc 

4- 11/),. 


Remarks. 


I'otato. 


+ 


+ 


+ 


+ 


+ 


lOacli reaction was 
intense. 


Root of 
Ipomaea 
Batatas. 


+ 


+ 


+ 


+ 


+ 


(d) was weaker than 
the other reactions. 


Root oi 

Raphanus 

sativus. 

(radish.) 


+ 


-+- 


+ 


^- 


'I' 


(a) Ljavc tlie weakest 
reaction. 


Root of 
vVrctiuni 
Lappa . 


4- 


+ 


+ 


+ 


+ 


(i1) gave the AveakcsL 
reaction. 


Apple. 


+ 


+ 


+ 




+ 


(c) was weakest. 


Fruit of 

Diospyros 

Kaki. 


+ 


+ 


+ 


- 


- 


(a) gave the m eakest 
reactioji. 


Bamboo-bhoot. 


+ 


+ 


+ 


+ 





All reactions were 
intense. 


Rhizome of 
Balanophorai 

Sp. 


+ 


+ 


+ 


+ 


+ 


(a) and (b) were 

stronger than the other 

reactions. 



In these tables + means a positive, and — , a negative result and o, that no test was made. In 
the comparison of these tests, it must be kept in mind, however, that the acidity of the juices and the 
presence of certain compounds inlluerice the intensity of the colorat'on. 

* This is a phanerogamic parasite. 



1 



On Oxidizing: EnxyiiiH in the Vegetable Body. 



21 



Longitudinal 

section 

of 


a. 

Guaiac 

tincture 

alone. 


b. 
Guaiac 
tincture 
H-II^O,. 


c. 

Guaiacol 


d. 

Parapheny- 

lendiaminc 

+ U/K. 


f. 

Tetramethyl- 

paraphenyl- 

endiamine 

alone. 


Remarks. 


Kesliug liarley. 


+ 


+ 


+ + 


+ 


The last reaction w caker; 

all reactions appeared 

only in the embr)o. 


Resting wheat. 


+ 


•i- 


+ 


+ 


•!- 


" 


Resting rice. 


+ 


+ 


+ 


+ 


+ 


•• 


Resting 
soy-bean. 


- 


+ 


-]■ 


+ 


+ 


" 


Germinated 
barley. 


+ 


+ 


+ 


+ 


+ 


ITie first reaction (a) wa-s 

stronger and the fifth (f), 

weaker than the rest. 


Germinated 
■wheat. 


+ 


+ 


+ 


+ 


+ 


" 


Germinated 
rice. 


+ 


+ 


+ 


+ 


+ 




Germinated 
soy-bean. 


+ 


^- 


+ 


+ 


+ 


" 


Aspergillus 
oryzae on 
boiled rice. 


, - 


- 


- 


+ 


- 


The violet color with 

(f) appeared along the 

rim, where the fungus 

developed. 



The plumules of the germinated plants were about i cm. lonij ain.1 the 
tests obtained showed the distribution of the oxiding enzyms to the whole 
extent of the sections. 

The tests obtained leave no doubt that the oxidase which gives a 
blue reaction with guaiac tincture (laccasc .') increases during germination 
while spcrmase decreases, which is in accordance with tlie observations 
of Griiss, that the reaction for spermase which at first increases, decreases 
later on, until it fails altogether. 

The second series o^ tests were made with various vegetable juices. 
The objects were cruslicd in a mortar with addition o( some quartz sand 
and extracted with some water. The filtrates bch ived as follows : 



2l6 



K. Aso ; 



in 

£ 


11 

— rt 


c 
1) 

1 


o 
o 

c 


I 


c 
o 

C 

_o 

o 

a 
P 


In testing with (a), a few drops of 

guaiac tincture are not sufficient 

to produce a blue color. 




H a 


+ 


+ 


+ 


-h 


o 


+ 


+ 


c 

CI. rt i-i 
ri 


+ 


+ 


+ 


+ 


+ 


+ 


+ 


. 1 < 

o + 


+ 


+ 


+ 


+ 


+ 


+ 


+ 


a 

1 d* 

1 5 

a 
O 


+ 


+ 


+ 


+ 


+ 


+ 


+ 


a. 

Guaiac tincture 
alone. 


+ 


+ 


+ 


+ 


+ 


+ 


+ 


U-l 

O 

.2 8 


>> 

%: 

m (J 


S s 


rt 
'rt 


' 


C D 

< c 


1 


:; 


O 

3 
•— > 


o 

p. 


3 

o 
o 

(A 




'3 . 


o 
o 

1 

rt 


r/ 

O 

d 
> 
rt 


a 

« 

o 
> 



i 



On Oxidizing En/yms in tlio Vogotalile Body, 



217 





3 

X 



■■g 

(Li 


■ - 


S 
1? 




'J 

X 


'■J 

X 


J -^ 
y — 

7. 


+ 


+ 


+ 









u 


-i- 


+ 


+ 


+ 





1 


1 


+ 


+ 


+ 


+ 


1 


1 


■= 


+ 


+ 


+ 


+ 


! 


1 


'■J 


+ 


+ 


+ 


+ 


1 


1 


1 


+ 


- 


>- 
< = 


r 




= 


1 2 

S 3 


/■J 
'c 
> 




1— > rt 




x 

p 


^ 


§ 



^ 




•^ 


X. 


















X 




rt 


X. 


u 




.'^ 


u 





















. 


£ 


— 










u 













y 







,"* 




y 


I't 


j^ 


>s 


■^ 


y 






u 
y 


hf 


rt 


u 


> 


> 




•~" 


; — 


U 


y 


X 


- 


„_, 


> 




c 










= 


2^ 


u 


■g 


'.~ 


uT 




w 














X 






•^ 





— 












^ 


3 

£ 


if 


X 




3 


y 










u 


r^ 


:i 





— n « 



2l8 



K. Aso: 



Also an animal secretion, saliva, was compared in regard to these 
tests with the following results. The samples came from different 

persons. 1 





a. 


b. 


c. 


d. 


e. 




guaiac 
tincture. 


guaiac 
tincture. 
+ H,0, 


guaiacol. 
4H,0, 


paraphenylendiamlnc. 
+ H,03 


tetramethyl- 

paraphenylendianiinc 

+ H,03 


I. 


- 


trace. 


+ 


trace. 


trace. 


11. 


- 


- 


+ 


o 


o 


III. 


- 


- 


■i- 


o 


o 


IV. 


trace. 


- 


+ 


- 


o 


V. 


trace. 


- 


-f 


trace. 


trace. 



While the quaiacol reaction (c) was intense in every instance, the 
fourth (d) and the fifth reaction (e) appeared merely in traces. The guaiac 
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 tl)an saliva. Tlie 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 icsin and hj'drogen peroxid. We have therefore to distinguish two 

1 Control tests were made with boiled saliva, and in no case a coloration sets in. 




Oil Oxidizing: Enzvms in tlio Vosrotable Body. 



219 



kinds of peroxidases, one that gives tlic red reaction wiih guaiacol and 
hydrogen j^croxid, the other that gives a blue with guaiac lesin and 
liydrogen peroxid. It seems furtlier probable that there exists a third 
peroxidase^ which gives a violet coloration with tetramethylparapheny- 
lendiaminc 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 th-ere exists a separate peroxidase that gives 
the red guaiacol test in saliva, there is no proof th.at the common peroxidase 
does not give both reactions at the same time, since thus far no object 
was observed that would give tlie 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 Loetc- 
in his ' Ph)'?iological Studies of Connecticut Leaf Tobacco.' Since those 
influences have not alwa}-s been paid careful attention to. the existing 
discrepancies between the data obtained can hardly surmise us. The 
followinsf table shows the various data obtained thus far : 



* It seems to me very prohahle th.it the reaction with paraphenylendiamine and hydrogen 
I eroxide, and that with tetramethylparaplienlendiamine and hydroi:;en jeroxid mit;ht l»c causcil by the 
same enzym. 

2 Report ot" r. S. Oepartment ol" .\L;nc., No. 65. 1900. p. ::i. 



220 



K. Aso: 



Name of the 
enzym. 


Temperatur 

which kills the 

enzym. 


Duration of 
Heating. 


Remarks. 


Name of the 
author. 


An oxidase in 


63^C. 






Yos/iida. 


Urushi. 






An oxidase in 


above 70'^C. 






BcrlfiTiut 


Urushi (Laccase). 






Tyrosinase. 


below 7o"C. 




Injured at 
.So°C. 










72^C. 


4 min. 














ss°c. 


il hours. 












Oxidase in the stalks 
of the 


6o°C. 






Raiiborski. 


sugar cane. 






Oxidase in a tol,acco 
leaf. 


6r)--67X. 


3 m:n. 


I part dry leaf 

was extracted 

\vith2opartsH^O 


Lo,-u>. 


Oxidase in tea-leaves. 


76— 77''C. 


5 min. 


The solution 
was neutral. 


The writer. 


Peroxidase in the stalks 


95°^'- 






Racil'orsl-i, 


of the sugar cane. 






Peroxidase in tobacco 
leaf. 


87=C'. 


^ min. 


1, 

Ihe reaction oi 
- the solution 
was neutral. 




Peroxidase in 33^,,' 
alcohol solution. 


7o"i'. 


a few seconds. 


Loc-u: 
'1 


Peroxidase in lo9u 
(NITJ3 SO, ' 


qfC. 


after a short 
time. 





i 



On Oxidizing Enzyiiis in llic Vef^otabl" Ikdj. 



221 



Name of tlie 
cnzym. 


Temperature 

which kills the 

cnzym. 


iJuration of 
IIeatin!.(. 


Remarks. 


Name of the 
author. 


Peroxidase in cocoa 


on boilint,^ 




Still active. 


////;/-, r. 


milk. 




Salive oxidase w^hich 

produces guaiacol 

reaction. 


Cj2%-. 




Trace of the 
reaction. 


Dtifoiiy. 




^-catalasc in a tobacco 


72°C. 


more than 
15 min. 




Loc-u: 


leaf. 






74°<^". 


very soon. 












» 


75°C. 


I see. 


The greater part 
was injured. 


" 


a-catalase in a tobacco 
leaf. 


75°C. 


5 "'ill- 


Kaint trace of the 
reaction. 


" 


» 


80" C. 


I min. 





•• 


An oxidase in milk which 

paves Storch's 

ffact io!i. 


Sc/C 









III order to decide whether the reactions caused by h\-drL>gen pcrc>.\i(.l 
with guaiac resin, c^uaiacol, parapheiiylendianiiiie, and tetramethylpara- 
phenylendiamine disappear at one and the same degree of temperature, 
the following tests were carried out. 

Potato-, banibooshoot-, barley leaves-, and radish root juice were 
heated for a short time to 8o°C or over with the following results : 

( + ) This forms a noticeable exception from tlio general rule that enzyms arc kilkxl Ih:!ow the 
boiling lu-at of water. 

.According to ////< '.07 <v peroxidase of pus is notdestroycd.it i;o^ when di>>olvc\l in a weak acid 
medium. (1,t semaine me<l. vol. iS). .\ similar observativ^n was made by Sfitz.r with jx^roxidasc from 
animal organs (I'lUig. .\rch. vol. 67, p. 615). 



222 



K. Aso ; 







o 


o" 




u 


d 
















o 


L^ S 


o 


s 


5 


o 


o 


"u 


CO fo 


c 






c 


" 
















jj 














^ 


























'0 


^ 












rt 


U 












>^ 


ro .- 


o 


o 


Cj' 


o 




n 


=f S 


o 


o 





5 


Tf 


J. - 


= 


'^ 


= 


- 


■^ 


~, 


C<3 












>^ 














3 














> 




o 














CJ 










"c 
c 


,. ^ 


ri 


ti 


t/. 


tx 


i/. 




-^ 


O 


1 


5 


5 




c/n ^ 












rj; 


''- IT) 


tf. 


c/". 


tr. 


x 


-'■ 


•~ 




i^ 














y. 










•j; 


























(—4 














o 














o 


•V r^ 


rt 


ti 


ti 


bi 


tx 


.y 


^ ;S 


^ 










'5 




;tt 


5 





5 


5 




-«- CO 


^. 




IT, 




















>> 


^ 










-»:; 


o 




o 


CJ 


d' 


cj" 






O^ tH 




o 




c 


3 


rt 'CJ 


vo ^ 


5 


p 


5 


o 




^ rt 


tw ^ 


s 


■^ 


- 


E 


o 


t£-^ 












c 


r^ rt 














5 > 

O IS 


























."1 


^' 1 
o ^ 


5 




5 


5 


_CJ 


5 


:§ .^ 




1 






"o 


1 — i 






" 






"" 


c^r 


u . 












o 
o 


So .S 


i; 


o 


CJ 


Cj' 


Cj' 




1 <■ 


5 


S 


o 


o 


c 


t/j rt 










r^ 




s- 


tL ^ 


" 


^ 


^ 




" 


O 3 


CO 












^ O 














C C 














C 














rt -*3? 














^ s 




u 








D >^^ 


^1 


cJ -^ 




b/j 
5 

in 


o 


1 


'Sri 


'5 
1 — 1 


^ IT) 


"tr. 


5 






fS . 














[5 o 


<5 


o 


p 


p' 


o 




o ■> 


•J 


c 


CJ 




5 


3 
o 


P •; 


"o 










c 




CO 




























. — • U-t 






c 








c ° .• 




o" 


.c 








tMi 


p 


rt 




' 


'■ 






■^ 


'?. 


p 








o 


O 

o 


C 




5 










O 


r- 


&i 



:: s 



p -i-; 
CJ S 



p Ci. 



o 3; 

3 oj ^" 



-3 1^ c 






Ol! Oxiiliziii^ Kn/yiiis iii tli" V<'g<'tabl(^ lUniy. 



2^3 



The influep.cc of concentration upon tlie killing temperature w^is 
observed with the enzyms of cured tobacco-leavesJ A leaf of cured 
tobacco (5 g.) was extracted witii 100 c.c. water, and tested with the 
following results : 



Degree ami 
duration ol 

IicatiiiL;. 


So'C, 
5 -'^'i"- 


5 Min. 


5 Mill. 


S6— 87''C, 
5 Mil). 


90'C, 

5 Mill. 


' )xi(las(.'. 


trac'.-. 


i;o:ie. 


ij>:!e. 


,:.v:.,-. 


r.o';e. 


Peroxidase. 


stronLj. 


weaker. 


weaker. 


w caki;i-. 


Trace. 


Green react ion. 


slronL,'. 


weaker. 


^veaker. 


weaker. 


none. 


Violet reaction. 


stroni;. 


\\eakcr. 


weaker. 


weaker. 


none. 


Red rcacl'on. 


strong,'. 


weaker. 


weaker. 


wralscr. 


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 Sj^C for 2 minutes, the following result was obtained : 





Oritjinal ^-o!ution. 


a. 


i.. 


< )\!dase. 


none. 


none. 


'..O.U'. 


Peroxidase. 


trace. 


none. 


none. 


Green reaction. 


none. 


none. 


none. 


Violet reaction. 


none. 


none. 


none. 


Red reaction. 


strong. 


uio«lorato. 


xraco. 



Though the killing tcmpciatinc is not a constant magnitude under 
all conditions, nevcrthles.s these comparisons show \er\' clearly that the 
red reaction caused b}' !;uaiacol and hj'drogen peroxid is ilue to a separate 



* The rea.ction of the extract of tlie cured leaves was neutral. 



224 



K. Aso: 



enz\-ni which has more resistance-power towards heat than the otlier 
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 enz3'ms acting in absence of hydrogen peroxid, can be 
inferred from observations of ^r/'/r<:r//c/ and of ^i?///"^///^/!?/. The latter lias 
observed among other things that when the extract of the fungus Riissula 
delica was kept witli 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 b>- certain poisons or by freezing. 
Only hydrocyanic acid and h}-drox}lamine hinder the action. 80%" alcohol 
does not kill oxidizing enzyms, but a stronger one will injure them slowl}-. 
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^ 
v/ith that of other oxidizing enzyms : 

Salts of a strong acid or alkaline reaction injure this enz}'m while 
salts of a neutral reaction do not. A remarkable fact is the depression 
of the activity of the soluble or ;9-catalase by nitrates, while the euf^yin 
itself is not injured by them. In general, tlie retarding influence of a salt 
is increased with the amount of the salt. Sodium carbonate attacks 
/9-catalase slowly ; 2% sodium fluorid and 5^^ dipotassium oxalate solutions 
cause no injur}'. ^% potassium sulphoc}-anid and thiourea interfere with 
the catalysing action, but have no direct injurious influence upon the 
enz\-m. Mercuric chlorid acts very injuriously. While highly dilute acids 

* Boiirqvclot : J. B. f. Thierchcm. 26. i>. 886. Cf. also his observations on the potato (il)icl). 
2 Lol-:{< : C'atnlasc, an c.-.zym of general occurc-r.c.-. I'. S. Dept. of Agric, Report No. 86. 



1 



On Oxidizing- Enzyiiis in the Vigotablc Body. 225 

retard the action of catalase, dilute alkaline solutions promote it. W'licn the 
amount of a mineral acid in the solution reaches more than o:^%, catalase is 
soon killed. At the ordinary temperature, 0.\% 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 o- 
cat.ilasc slow!}', and kills ;9-cata]ase in two da)-s. While o.\% caustic soda 
causes no injur}-, \% of it kills catalase instantly. Alcohol above ^0% 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 : \% phenol retards the- action 
of the enzym ; ij% formaldehyde destroys its action in a very short 
time ; 0.^% nitrous acid injures the enzym considerably in one day. 
«-catalase shows considerable resistance to the action of hydrocyanic acid 
of 2%, while ,5-catalase is gradually killed by it. Afler the evaporation of 
hydrocyanic acid, regeneration of the action of y5-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 ^% 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. 

I. Influence of salts Jipon ilie color- reactions nientioucJ. 

5 c.c. of a dilute juice ^ of radish root were mi.xed with various salts 
and left with some drops of ether 'ior 48 hours before testing. 

In carr)ing out the o.xidase reaction i c.c. of guaiac tii.cture was 
added; for the green reaction, i drop of sodium acetiif. 1 tirL>p of para- 
phenylendiaminc and 2 drops of hydrogen peroxid ; for the red r« action, 
an equal volume of guaiacol solution and 2 drops of hydrogen peroxid ; for 
the violet reaction. 2 drops of an alcoholic solution of I'Y.o tctramethylpara- 

* It is ncccsfary to ililuto the juice n-.txlcratcly to obtain the color ro.;ction ot a degree s iitablc 
for coinparisoi). 



226 



K. Aso: 



pbcnylendiamine and 2 drops of hydrogen peroxid. In each case the 
intensity of the color produced was compared with that of the control 
solution without salts. 



Salt. 


Concentration. 


( )xyda'^e. 


Red reaction. 


Green reaction. 


Molet reaction. 


Sodium chlorid. 


5% 


weaker. 


weaker. 


weaker. 


\\eaker. 


Potassium 
nitrate. 


5% 


weaker. 


weaker. 


weaker. 


strong. 


jNIagnesium 
nitrate. 


5% 


sHghtly 
weaker. 


stro.ig. 


strong. 


strong. 


Calcium 
nitrate. 


5"o . 


" 


" 


" 


" 


^Magnesium 
sulphate, 


5% 


strong. 


strong. 


strong. 


" 


Ammonium 
sulphate. 


S% 


'■ 


" 


" 


" 


Sodium 
sulphate. 


5% 


" 


■' 


»> 


» 


Dipotassium 
phosphate. 


5% 


weaker. 


weaker. 


a violet color 
a]ipeared 
instantly. 


wealcer. 


Monopotassium 
phos]5hate. 


5% 


strong. 


strong. 


strong. 


strong. 


Sodium 
cai'ljonate. 


2% 


weaker. 


weaker. 


turned quickly to 
violet. 


slightly 
weaker. 


I'otassium 
oxalate. 


l"o 


weaker. 


weaker. 


not green, but 
violet. 


almost none. 


Ammonium 
oxalate. 


2% 


strong. 


strong. 


strong. 


weaker. 


Sodium 
fluorid. 


-.0 


none 


moderately 
strong. 


molerately 
strong. 


moderately 
strong. 


Sodium 
silicofluorid. 


0.3 "o 


none. 


none. 


none. 


none. 



On Oxidizing: Eii/ynis in the Vcgrcfable 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 enz}'ms, or weaken at least the color reactions caused I.y tiiem ; of 
special interest is here also the influence of potassium nitrate, since this 
depresses the action also of cataiase, without injuring the en7.)-ni itself.^ 
Sodium carbonate {2%) and potassium oxalate {i%) weaken also these 
color reactions. The influence of sodium chlorid {$%) and potassium 
nitrate ($%) 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 th.e case of chlorid of sodium and in regard to the 
red reaction. 

The striking influence of sodium fluorid and of sodium siiicofluorid on 
the appearance of the color reactions led me, not only to repeat the tests 
with radisji juice, but to make also some further tests ; ^.^ grms. c f a'r dry 
tobacco leaves were crushed in a mortar and extracted with 300 c.c. water. 

The filtrate served for the followincf tests : 





Concen- 
tration. 


Oxydase. 


Peroxydase. 


Red 
raection. 


tlreen 
reaction. 


\-iolet 
reaction. 


Time of testing. 


Sodium 
tluorid. 


2/0 


trace. 


weaker. 


weaker. 


weaker. 


weaker. 


Immetiiately. 


" 


^0/ 
-/o 


none. 


weaker. 


w eakei . 


weaker. 


weaker. 


After al>out 
one hour. 


'• 


5/0 


none. 




■' 


•• 




Immeui.itcly. 


Sodium 
siiicofluorid. 


-/o 


none. 


weaker than 

in tlie case 

of XaF. 


weaker 
than in 
the case 
of XaK. 


weaker 
tlian in 
the case 
of NaF. 


weaker 
than in 
the case 
of XaK. 


•« 


•• 


->0' 
-^0 


none. 




- 






After about 
one hour. 


" 


5% 


no!ie. 






•• 




Immoli.ilely. 



* 1. c. ZctTi' : latakise, an cn/yui of ijeneral occurer.ce, I". S. IVj t. of Agric. Kcixv.t No. 6S. 



^28 



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 (luorid 5% 


none. 


weaker than control. 


tested. 


Sodium silicofluorid 0.3% 


none. 


slight. 


immediately. 



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. Inftuerice of dilute acids and alkalies 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 slightly, 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 grms. 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. ArtJiiis and 
///^/'(V, Jahresb. Thierchcm. 1893, p. 640. 



On Oxi(li;:iiii? Eiizyiiis in the V«'ij;ctal)l«' Body. 



229 



Acids or 
alkalies. 


Concen- 
tration. 


Time . , , 

I- • • ' ixv( asc 

aner )nl^■l^!^^ ^ • 


Peroxydase. 


! 

Red Crcen \*io!ct 
rcaaion. reaction. reaction. 


Hydrochloric 
acid. 


,0X 
*/0 


5 hours. 


none. 


none. 


trace. 


trace. trace. 

1 


Nitric 
acid. 


1% 


" 


none. 


none. 


none. 


none. 


none. 


Sulphuric 
acid. 


1% 


" 


" 


" 


" 


! 


vKcctic 
acid. 


2% 


" 


» 


" 


" 


i 
1 

\ 


Oxalic 
acid. 


-,0/ 


" 


" 


" 


- 






Caustic 
soda. 


1% 




•' 


" 


- 


1 


" 


Tartaric 
acid. 




24 hours. 


1 


" 1 ' 

1 


1 

! 





This was repeated with th.e water extract from a cured tobacco 
leaf with the difference that the tests were made 2—3 minutes after 
mixine : 



Acids or alkalies. 


Concen- 
tration. 


< >x\Hlase. 


IVroxydase. 


Revl 
reaction. 


Gixvn \'io!ci 
rcaaion. rcactioi\. 


llydrocliloric acid. 




none. 


trace. 


trace. 


nop.e. j none. 

1 


» 


1.%' 


trace. 


trace. 


weaker. 


trace. weaker. 


'I'artaric acid. 


^% 


weaker. 


strou!^. 


stroni:^. 


strong. 


stron«j. 


Caustic sola. 


^?o 


none. 


none. 


none. 


none. 


none. 




l"o 


weaker. 


weaker. 


weaker. 


1 
w eake:". \\ eakv r. 



230 



K. Aso; 



3. Iiifiiience of Poisons. 

Influence of li}'drocyanic acid : Two leaves of cured tobacco (about 
10 grms.) were extracted with 200 c.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 tlie 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 immediatel}' afterwards, all reactions were still 
obtained. But, v/hen 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 alcoliol was added to precipitate the enzyms and separate them 
from phen}-lhydrazine, since this might have interfered with the color- 
reactions. The precipitate was collected in a Alter, 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 witli the actions of nij-rosin and emulsin. Recently, 
Hunger^ observefl that a reducing sugar present in the milk of the cocoa- 

* Cf. alio E/stciti, Archiv. f. J lygicnc, vol. 36 [\ 140. I'russic acid prevents the coloration of beet 
juice cxiioscd to air, while after expulsion of that acid by a current of air this oxidative process sets in 
a^aiu. 

* Ihmga- expressed in a recent article (lier. Deutsch. Bot. Ges vol. 19, 1901), the supposition that 
in my tests for oxidase and peroxidase in ripened and unripe kaki fruit?, it was the sugar and not the 
tannin wjiich interfered with tlie reaction. This is however not so. 



Oil Oxidixiiii? j;iizyiu> in tiic \('!4:t'tiil)l(' Hod.v. 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.' I 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 100 c.c. of a dilute radish juice, 10 grams of ligg albumin or 
pepton were dissolved and this mixture tested after 24. liours. 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 x'egetable enzyms have been studied 
but little. It is ah-eady known that there exist zymogens of pepsin, 
trypsin, rennet and diastase in the p.nimal bod}-. As for vegetable enz\-ms. 
zymogens of a proteolytic cnzym in Xcfctil/ics and lupin-seeds, of inulase 
in the resting tuber of artichoke, of lipase and rennet of the castor oil seed 
and of a diastasic enz)-m in barley and wheat have been observed. - 
Thus far, however, zymogens of oxidizing enzyms have never been 
investigated, although thes' must exist, as the following obser\-ations will 
show. 

I. juice of barle)- was heated to 8o"C. for 5 miiuites, whereupon the 
oxidase reaction failed to appear, the oxidase having been changed. 
After standing for 24 hours, the oxidase reaction was again obtained. 
Another portion of the same juice was heated to 85~-C. for 5 minutes, where- 
upon no oxidase reaction, but a trace of peroxidase antl the red reaction 
was obtained. After 24 hours standing, a weak oxidase and a strong 
peroxidase reaction were again observed. The reaction of the barley 

^ I/iin^ii-, llel oplivd(.n ikr o\yi.la>crc.iCtio in vcrlMiul mot »lc IvXMli-vitic dor v;ly(\'>^c in hot 
suikcrict. Archicf voor de Tava-Suikorindustnc, 1901. 

* Cf, K, Gn-iii : The SoluMo Kcrments and 1-Vrniont.ition. p. v'^l 3')i- 



232 



K. Aso: 



was very faintly acid. Similar observations were made by Albert 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 liours standing. But 
when, after the boiling, \% 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. Loeiv 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 : no regeneration. 

,, 48 ,, peroxidase, red and violet 

reactions appeared slightly. 
,, 4 days the same. 

5. The same tests were carried on with milk which was boiled for 
a few minutes : 

After 24 hours : no regeneratio'.i. 

,, 48 ,, all original reactions 

reappeared, but weaker. 
, , 4 days the same. 

6. The water extract of cured tobacco leaves boiled for a few 
minutes was tested as follows : 

After 24 hours : n(j regeneration. 

,, 48 ,, trace of peroxidase and red reaction 

reappeared, the latter stronger than 
the former. 

1 Also this iiutlior linlcnvd llic existence of corresponding zymogens. These facts as \\ell as the 
observations of S/o-ivtrA^ff (A. pliysiol. Cheni. 31.) and TT/.f/./;/ (Arch. ITyg. 30.) seem to fully estabh'sh 
the enzym nature of the oxidases. Recently, however. Kastle and Locivenhart (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 cnz 



Oil Oxidizing Eii/jins in the Vegetable Body. 233 

After 3 days : Peroxidase and red reactions reappear- 
ed weak. 
,, 5 days : Tlie 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 : 100 c.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. 

Control 29.1 c.c. 30.9 c c. 

Heated 4 c.c. 4.8 c.c. 

" Regeneration " of Catalase seems to have taken place in a 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. Loezv has already shown that 
oxidase and peroxidase are not nucleoprotcids, 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 mi.xed 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 ^' ^^'^*^' 

Upon addition of three volumes of absolute alcohol to the radish 
juice, the filtrate failed to show the reactions, while tlie 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 
BeJirens with th.e juice of tobacco-leaves. Upon mixing with double the 
volume of strong alcohol a filtrate was obtained, which gave onl}' the 
peroxidase reaction. 

All oxidizing enzyms tested are precipitated by saturation vi'ith 
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 i% acetic acid to the radish juice, 
a small quantit}' of a white precipitate was obtained, but all reactions 
could be obtained with the filtrate. Hence tlic substances which cause 
such color reactions belong in all probability to kinds of albumoses and 
not to the nucleoproteids. 

Anotlier method of separating peroxidase from oxidase may be based 
on tlic 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 attaclced by sodium fluorid and sodium 
silicofluorid.* 

General Conclusions. 

1. Various vegetable objects which }-ield the well known guaiac 
reactions for oxidase and peroxidase, also yield a red reaction 
with guaiacol and hydrogen peroxid. 

2. Storcli s reaction on milk with paraphen)'lendiamine 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 1 used 5% solution of sodium iluorid and a saturated solution of sodium silicofluorid and tested 
after well shaking the mixed solution. 



1 



On (^xidiziiifr Knzyins in tlic >'(■>;«•( able IJody. 



235 



3. A new reaction for an oxidizincr cnzym was found, wlu'ch consists 
in the production of a deep violet color on the addition of tctrame- 
thylparaphenylendiamine and hydrogen peroxid. This reaction is 
obtained with various vegetable objects. 

4. This new reaction may be used to distinguish fresh milk from 
toiled one. 

5. The spcrmase reaction found by Grilss is also obtained with several 
seeds, resting as well as gerniinated ones. In the case of resting 
seeds, only the embryo j-ields this reaction. 

6. The red guaiacol reaction is caused by a separate enzym which is 
more stable then even the peroxidase.^ 

7. Sodium fluorid and sodium silicofluorid interfere with all the color- 
reactions. Oxidase is killed sooner than the other oxidizing 
enzyms. 

8. The green and violet reactions might be caused b\' enzyms different 
from oxidase and peroxidase, since 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. 

9. Sugars do not interfere with the color reactions caused b)' oxidizing 
enz}'ms in aiu- notable degree. Neither interfere soluble egg 
albumin and pepton. However t.iniu'n interferes \'ery scricaisly. 

10. The presence of z\'mogcns of the oxidizing enzyms is ver\' 
probable. 

II The separation of peroxidase from oxida>^e miy be accomplished 
by adding two volumes of absolute alcohol to one volume of a 
plant juice. Hereby oxidase is precipitatet! while the main part of 
the other oxidizing enzyms is present in the filtrate. 

12. The oxidizing cnzN'ms which i)roduce these color re.iclions resemble 
albumoses. 



' 'lliorc sooius to exist a rolatal oxid.isc wliich pro Uici"> t1io s.iiiK' iv.'.v'ion without hydrogci» 
licroxu! as /u>//n/iti-A>/ has oliscrvod wiih AV/.o-///./. 



L 



On the Curing of the Kaki Fruit. 



BV 



S. Sawamura. 



The fruit of Diospyros Kaki L. is an article of food extensively 
consumed in Japan. The flesh of this fruit contains glycose and fructose, 
while the re.-erve carbohydrate in the seeds is mannan.' The fruit was 
found to consist of :- 

Sweet variety. Astringent variety. 

Water 82.03 83.65 

Nitrogenous matters 0.61 0.5S 

Fat 0.02 0.02 

Sugar and other N free extract . . 13.62 12.56 

Fiber and kernels 3.29 2.76 

Ashes 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 011 by oxidizing enzyms which act on the 
tannin as Aso^ has shown. As to the astringent variety artificial means 
arc resorted to, to remove the unpleasant taste. These means are : 

I. Keeping the fruits for 12 hours in a barrel containing vapors of 



« /.*////. These liuUotins, \'oI. II. Xo. 2. 1S96. 

^ Acconling to the analyses of the Tokyo Sanitary Exp. Station. 

=» GiH'cr found that wIumi this fruit is allowal to ripen in a continetl atniospheiv, tt yields 10% 
of ethyl alcohol, mixed with other alcohols, anions; which aniyl alcohol was copious, and further tlut 
thcaiiMnatic princi[ile was a mixture of aniyl and ethyl acetates w ith traces of ocn.inthyl.-itcs and 
pelargonates. [GriCii : Fermentation p. 351.) 

■• I'lOtanical Maijazine. Tokvo. icfoc. 



238 S. Sawamiira: On the Curing of tlie Kaki Fruit. 

alcohol. Generally Sa^i'-ha.rre]s just emptied are used for this 

purpose. 
II. Keeping- the fruits for 12 hours in warm vvater.^ 
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 tliat 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 sh.owed that the change did not consist in tlic trans- 
formation of tannin into sugar. ^* Tjierc 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 ox}-dation brought 
on by the oxidizing enzyms present \n the fruits. These emzyms are 
confined to the cytoplasm, wlu'le the tannin to the vacuole. By killing 
the cells the osmotic properties of tlie cytoplasm are changed, and the 
oxidases can now i)ass into tlie vacuole, wliere they can mix with the 
cell sap and exert their action upon the tannin. 

1 30^-40'' C. are sufficient to t-ficct the change. 

2 For this purpose a ripe fruit, whicli liad still an astrini^a^nt taste, was divided iiito two equal parts, 
one of ^\•hich ^\-as cured by exposing to vapors of chloroform, v/hilc tlie other left viithout any treatment. 
Ill the former 55.91% of surar but no tannin was found, while in the latter 56.18% of sugar and 8.23% 
of tannin were containe<!. 



On the Different Forms of Lime in Plants. 



BY 



K. Aso. 



In the quantitative determination of mineral constituents of i)lants 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 i) 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 extr.icted by dilute 
acetic acid but not by boiling water. 

Since the juices of plants have generally a more or le^-s 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. 

IMie [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 selected : 



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. 1900). 

3. Wild clover. (Leaves and stems, collected before flowering, on 
Apr. 26. 1901). 

4. Barley. (Leaves and stems, collected before flowering, on Apr. 26. 
1901). 

20 grams of air-dried finely powdered substance were extracted twice with 
one liter of boiling water, ^ and the residue thoroughly washed with hot 
Vi-ater until every trace of soluble lime was removed. This residue was 
extracted with one liter of ^% acetic acid in the cold for 24 hours, 
frequentl}' 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 c^% 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 ICO parts of dry matter, 


Objects. 


Ca( >, soluble in : 


Total. 




Water 


Acetic acid 


Hydrochloric 
acid. 


I'olato 

Buckwheat 


0.332 
0.056 
0.85 S 
0.43^ 


0.875 
0.367 
0.742 
0.259 


1.5S6 
1.524 
0.489 
trace 


2.793 
1 .947 3 


Clover 


2.089 


IJarley 


0.697 



1 The aqueous solution had a slight acid reaction in each case. 

■^ In the final residue, lime and ma<i;ncsia were absent or present only in very minute traces. 

" The- liire factor - for buckwheat hefori.- ilowerini/ is -;, while in the time of fruitintr it is !.•?, 

Mi.;() - J' 

V hich show s that nia<:;ncsia plays also here a very imjortant rule in the fruitintr process. 



On llic IMlIVrciit FoiJiis of Limf in Mant? 



241 





hi 100 parts of dry matter. 


Objects. 


MgO, solul>Ie in : 


ToLi!. 


. 


\\'ater 


Acetic acid i Jlyl'ocbloric 
t aci<l. 

1 


Potato 


1.617 ^ 
1.050 1 
0.491 
0.307 


0.550 
0.417 
0.162 

0.094 


0.217 
trace 
trace 


2.3S4 
1.467 
6;'; 


Kuckv, heat 

Clovei" 


• Rxrley 


0.401 



We see from this table, that the quaiitities cf the difiereiU forms of 
lime vary considerably. In the case of potato and buckwheat, only a 
small quantity of lime compounds soluble in water are present; nnue lime 
is found in the acetic extract and still more in the hydrochloric acid 
extract. Much calcium oxalate is produced in these leaves, while th^sc of 
barlc)'- 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. 

CJiuicJis investigation^ with albino leaves have demonstratetl that 
lime is more abundant in the green leaves than in the white. In order to 
see whether this pathogenic albinisu"! shows a chemical analogy to the 
normal albinism, I have determined the lime also in th.c white and grtcn 
parts of the !ea\-es of Ariindo Doiiax .^eparatcl}".^ 



> I'robably present as secondary phosphate, in whicii form it is not iMDisonous for tlie nuclei. 
I have made also analogous determinations of lime in maize-stalks with the following rt-sult : 

In ICO parts of dry matter; soluble in 

Water. Acetic a c'd. Ilydi-ochloiic acid. 

CaO 0.130 0.12S trace 

ISIijt") 0.241 0.120 lr.'.ce 

l?ut, as these stalks had been dried uid werc exiio.-e<! to the rain on the tield af'er harwstini:, the 

result is not a normal one. 

2 (."alcium oxalate is absent in most Grii'iiiiii.c. 

3 Journ. Cl'.em. Soc. 1S7S and 1S86. 

* This separation by scissors was made as carefully a^ ) os>ib!e. nevertheless it was not alsolutely 
complete. 



242 



K. Aso: Oil llie Dift'ereat Forms of Liiiie in Plants. 



The analysis was carried out in the way above mentioned. 

In 100 parts of dry matter, total ash : 

White parts 11.83 

Green parts 14-47 





CaO soluble in 


Total. 




Water 


Acetic acid 


Hydrochloric 
acid. 


White parts 

Green parts 


0.213 
0.313 


0.216 
0.226 


trace 
trace 


0.429 
0-539 





.MgO solul)]e in 


Total. 




\\'atcr 


Acetic acid 


Hydrochloric 
acid. 


White parts 

Green parts 


0.387 
0.444 


0.068 
0.069 


— 


0.455 
0.513 



Lime- factor : 



White parts. Green parts. 
0.9 I.I 



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 
factor vr— 7=A in the grreen parts is larc^er than i, while that in the white 
MgO ** 

smaller than i. Hence the inference that the amount of lime increases 
with that of the chlorophyll bodies, other things being equal, has again 
found confirmation. 



On the Alcohol Production in Phaenogams. 



BY 



T. Takahashi. 



Since the interesting discovery oi E. Biichncr that the expressed juice 
of yeast can cause the alcohoh'c fermentation of glycose and fructose, the 
question lias been raised, whether zymase is also produced in phrenogams, 
and whether the alcohol production in the process of intramolecular 
respiration of cells of phrenogams is due to zymase or to the action of the 
protoplasm itself. Former observations made by Bre/eiiP were not in 
favor of the assumption of zymase. He had observed that the expressed 
Juice of grapes the surface of which had previously been sterilized, did not 
show any alcoholic fermentation while the sterilised intact grapes them- 
selves formed some alcohol by intramolecular respiration, like man\- fruits 
rich in sugar do. 

Recently Godlcii'ski published an cxhausti\e in\-estigation on the 
intram.olecular respiration of the pea.- lie 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, G oJ I eic ski ^xow\\6. the peas to a fine 
powder, wliercb}' the protoplasm would be killed, but ZN'masj cm remain 
intact. In this condition hardi}- a trace of carbonic acid (and consequently 
also no alcohol) was formed during two days, ' while the same weight oi 



' Lanchv. Jaluh. 5.(1876). 

2 Hullctiu (Ic rAcatlemio dcs Sciences de Cracowic, 1901. 

* The I ea seeds seem to pitxluce more alcohol tlian many other stxxls. G<.\iit':vski mentions : .. Die 
Meiige lies AIcoliols, w eleho W\ der intra molccul.u en .Vtlununvj der in rcinen\ Wasscr liegcnden Erbscii- 
sainen sich bildet, kann his i\\ 22% der ui-spriinglichen Tivckensuhstanz der Samen envichcn." 

■» Later on bacterial action was nolicetl. 



244 



T. Takahashi; 



entire seeds produced 5 cc. carbon dioxid in 24 liours. But although tin's 
result apparently proves the absence of Z3'mase, Godleioski hesitated to 
draw this inference, and declares among other thint^s : ,,Es ware moglich, 
dass durch das Zerreiben der Pflanzenmasse irgeiul welche Substanzen 
aus gewissen Zellen frei gemicht werden, welche, sci es durch Nieder- 
schlagen der Zymase, sei es auf andere Weise, die Wirkung derselben anf- 
heben." 

I have made the following experiments in re;:jard to the intramolecular 
respiration of the pea. 

One hundred seeds, weigliing 33-3795 g"- in the air dry state were 
left for one hour in a i per mille solution of i-jicrcuric 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 m.ostly 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 developm.ent \\ as continuous with the exception of those days, on 
which the temperature had sunk to ^. During the 38 days of observation, 
the water above the peas Iiad 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 i7,5°=0.99646. The iodoform test 
proved further that the alcohol was iiideed etliyl alcohol. This result fully 
confirms Godlezvskts observation. The quantities of alcohol, however, 
were larger in Godleivski s experiment, what can be accounted for by the 
higher temperature (17,4-24,7°) in the latter case. 

A further test proved that a number of the peas from the flask 
had still retained their germinating power. The water, further, in which 
the peas had remained and from which the alcohol had been distilled 
off ser/ed for the determination of the solid matter it had extracted from 
the peas. The residue weighed 1.146 g.=4,oij)^ of the dry matter of the 



Oa ilie Alcoliol rrodiictioii iu Plia'iio^ams. 



245 



peas.' (^f this nearly one fourth consisted of mineral matter. 

In order to decide whet'.ier 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 
3i*^C. Kvcn the smallest quantities of z}'iTiase would thus have been 
betrayed b)' some development of carbon dioxid, but 7iot a single bv.hble 
luas noticed even after one day. I must infer, therefore, that .ryiiiase is 
absen.t TxwCi cannot be tJic cause of the alcohol production in the intrmnole- 
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. Godleivski arrives at the 
view : ,,Die intramoleculare Athnunig im Sinr.e der alcoholisciien Giirung 
bildet unter normalcn Bedingungen aller Wahrscheinlichkeit nach das 
erste Stadium der nor.malen Athmung in alien denjenigen Fallen, wo sich 
dieselbc auf Kosten der hydrolysirbaren Kolilenhydrate vollzieht." This 
view seems to mc not justified. In the first place we would have to 
assume two different ways of res[)iration 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 '\\\ regard to the chemical character of fat and su;::ar. The 
latter is much easier oxidized than the f.ir, 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 GodUivski^s 
view, that those seeds that can jiroduce more alcohol by intramolecular 

1 Tlic iVo.-Ii peas cont.iiuixl 2S.55J.V >^H" dry nutlor. 

^ It may ho nicntioncil here lh.it ^^lllc has fouml some alcoliol hi jca <ot.\llni;« thrif ha«l trcrmi- 
iiatcv] under nornial comlitions at 24'^". for 4S hour:?. 

Tlie view of if^/z-.v// that yymaso exi^ls in the \<c\ (4UO'.e\l l>y Gt\fHy VVnr.eVia;! n-^j iiiv.;* :iv"> 
support hy my experitnents. 



246 '^- Takaliaslii: On the Alooliol Prodiutioii in Pli^nogams. 

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 goes, lliere may be other products formed in other seeds 
from sugar, wlien 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 
0. LocicJ 

1 Cf. O. Locxv : Die chemisclie Energie dcr lebenclen Zellen (Chapter XII), Miinchen 1899. 



Can Alcohols of the Methane Series be Utilized as Nutrients 
by the Green Plants? 



BY 



S. Sawa. 



Wliile alcohols in moderate concentration act poisonously on the 
higher plants, as Tsiikainoto has shown/ it seemed to me probable that 
in proper dilution various alcohols might be a nutrient for them. It is 
known that methyl and ethyl alcohol occur in small qu.uitities in 
certain green i)lants. Methyl alcohol was observed by Gutzeit as well 
as by Maquenne in the distillate obtained from juices o{ Ei'onymus, Hedcra, 
Lolium, Uriica, Galniiii, HeliantJius, Syringa, Dalilia, Acorns, ffcraclcuvt, 
and Pasthtaca. 

IClhyl alcohol has been found in Heracleinit, Pastinaca and Authris- 
ciis. P. Ma::e 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.- He thinks that this 
alcohol was formed from glucose b)- a kind of fermentation process in the 
cells. It has been known long ago further that alcohol ma)- be formed 
in the interior of sweet fruits. It has also been shown by Bokorny 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 forniic aldehyde, the next 



* Journ.il of tin- Co'lci^'c ol" Science Tokjo, 1895. 

» C'onipaie also o<'i//<r>'.>Xv', Talnvsbcr. f. Thicrd'cmic 1S97, p. 700. 

* In darkness this process does not take phice. 



248 >». ^iv.ya: 

oxidation-product of inethyl alcohol.^ It seemed I0 mc of interest to 
compare, therefore, the action of methyl alcohol on th.e growth of the 
plant with that of some other hiuj-licr alcohols. For the following experi- 
ments served young onion plants grown from the seed which were kept 
at first in o. !_%" solutions of methyl, ethyl, butyl, and isobutyi 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 ,, }ilagnesium sulphate, 

24 ,, Potassium nitrate, 

6 ,, 3.Iono-])otassium phosphate, 
trace of ferrous sulphate. 

5 cc. of this mixture were added to lOO cc. of the 0.1% solutions of the 
alcohols on the 26tli of ]\Iarch. The length of the leaves was measured 
at the same time and the solutions rciiewed 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. The letter "o" in 
the table siginTies the leaves present at the starting of the experiment 
while the letter "n" signifies the new leaves formed. The temperature 
of the room varied between I2-22^C. Tlie flasks containing the phmts 
were placed on a table well ex[)osed to tlie diffused day-light, but not to 
the direct sun-light. ~ Some of the leaves dried off gradually at the 
tips andthese dricd-off parts were not considered in the measurement. 



1 KinosJiita lias further found lliat mclliylic alcohol can be used by green plants for the formation 
protein (I3ul. College of Agriculture, Tokyo 1895 vol. IT. Xo. 4.) I.oc7o had observed before, that it 

can be used by bacteria as food, even in absence of oth.er organic inaterial. 

2 Perhaps the nutrient effect of assimilation under very bright light would have obscured the 
nutritive effects of methyl alcohol. 



Can Alcohols of tlic Methane Series be Utilized as Nutrients by the Green Plants .' 



249 



Alcohol. 




Length on March i6th. 


Length on March Increase in absence of 
26th. mineral nutrient?. 




Individual 
leaves cm. 


Summed up 


Individual 
leaves cm. 


Summed Absolute 
tip cm. 


Relative ^^ 


A 

Motliyl alcohol- 

1! 


21.5 
0' 1 1.2 


H 


23.0 
0' 18.0 
n 1 0.0 


5T.O 


1S.3 


55-9 


" 19-5 
0' 19.2 


■38.7 


26.5 

"' 14-3 
n 12.5 


53-3 


14.6 


37-7 


Ethyl alcohol 


/A 
]! 


21.7 (i 
0' 21.5 


43-2 


7.0 
0' 2S.2 
n 12.5 


477 


4-5 


10.4 




24.S 
0' 13.8 (2) 
0" 1 1.6 


•50.2 


26.0 
0' dead 
0" 29.0 


55-0 


4S 


0.S 


Butyl alcohol 


/A 


16.5 
0' ,3.5 


30.0 


24.0 
0' n .0 


35-0 


50 


16.7 




1S.2 
0' 10.2 


2S.4 


'"> 22.2 
0' 13-2 


35-4 


7.0 


24.6 


Isobutyl alcohol. 


rA 
1; 


20.1 
^' 9-9 (3) 


30.0 


^^ 21.3 
0' dead 
n 20.0 


41.3 


"•3 


37-7 


1 


1S.2 
0' ,;.2 


35-4 


0' iS.o 


42.5 


r.. 


2C,I 


Control. 


A 


*^ »4-9 (4) 
'^' >3-5 


2S.4 


dead 
0' 26.S 
n 12.0 


3S.S 


10.4 


36.6 




1; 


20.5 
0' 11.2 


31-7 


24.5 
0' 16.0 


40.5 


V n; 


27.7 



250 



S. Sawa: 



Alcohols. 



Ix'iigth on April 24th. 



Individual 
leaves cm. 



Summed up 



Increase after mineral 
nntrients were given. 



Absolute 
cm. 



Relative % 



Remarks. 



Methyl alcohol 



dead 
30.2 
29.0 
25.0 
20.5 
"•5 (5) 
5-5 



121.7 



707 



138.6 



(5) A bud was formed 
on April 17th. 



dead 
26.0 

44-5 
32.0 

21.5 



124.0 



70.7 



25-5 
45 -o 
34.0 



104.5 



56.8 



119. 1 



(i) 1.7 cm on the tip 
had dried off. 



Ethyl alcohol^ 



20.0 
dead 

37.5 
35.6 (6) 



93 -o 



38 o 



69.1 



(2) 3.0 cm on the tip 

had dried off. 

(6) A bud was formed 

on April 18th. 



dead 

23-5 
29.7 
16.0 



611.2 



28.2 



80.6 



Butyl alcohol.^ 



dead 
24.0 
28.6 



52.6 



17- 



48.6 



Isobutyl alcohol 



21.5 
dead 
35-7 
23-5 



80.7 



39-4 



95-4 



(3) 5.0 cm on the tip 
had withered. 



dead 
dead 

33 -o 
27-5 



60.5 



18.0 



42.6 



dead 
29.0 
36.2 
5-o(7) 



11.4 



80.9 



(4) 3-3 cm on the tip 

had died off. 

(7) A bud was formed 

on April 20th, 



Control 



dead 

31-5 
370 
8.0 (8) 



76-5 



\G.o 



88.9 



(8) A bud was formed 
on April 19th. 



Can Alcohols of the McthaiK' Soii<'s bo Utili/ert as Nutrients by the Green Plants I 25 1 

The result is therefore that methyl alcohol in o.\% 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 addetl 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 tiie growth.^ 



1 The higher alcohols are also more i:oisonous tor plants than the lower as Tsukatiiolo had already 
observed. " 



On the Occurrence of Mannan. 

C. Kimoto. 



The peculiar horn-like consistency of the seeds of Trachycarpus 
excclsa, a palm tree, frequently servin- 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 o{ Diospyros Kaki.^ 

Recently Bourquelot and H. Herissey observed a considerable amount 
of. mannan in the seeds of Phoenix Canariensis. Of considerable interest 
is further the observation of Tsnji that the root of Conophallus Conyaku 
{Amorphophallus Rivicri), used m Japan as an article of food, is exceeding- 
ly rich in mannan. - 

Tsukamoto has shown further that tliis plant contains also mannan 
in the leaves and stalk."' Of special interest is the fact discovered by 
Gabriel BcrtrancP that while x>-]a.i is present in the wood oi Augiosprrvis 
it is replaced in the Gymuosperms by mannocellulose. The wood of 
Abies pectinata yielded 9.6^ mannose, on being boiled t^ve hours with 
hydrochloric acid of 5 0/ Gnetacecv which from the transition between the 
Gymnosperms and the Angiosperms gave thus no or very little mannose. 

In order to test the seeds of Trachycarpus excelsa for mannan. the 
seeds were finely cut up after removing the shells, and 2.07 g.(- 1.49 g. 

» Ishii. Hul. vol. lI.^Xo. 2 of r.n;.. C'olk-^v ol .\v;ricuUuro. I nivorsity of Tokyo. 

« Ihiil paiji- 103 and K'inosJiif.i, Il):d No, 4. 

» ll>i\I. vol. III. 

* <^'. v., vol. 130, p.iv^o 1023. 



254 ^'' Kimoto : On tlio Occnriv'uce of Mainian. 

dry matter) boiled for three hours with sulphuric acid of ^%, replacing 
the water lost by evaporation. The filtrate was neutralized with barium 
carbonate, the liquid evaporated to moderate concentration and acetate 
of plienylhydrazlne added, which produced a voluminous crystalline pre- 
cipitate. yXfter standing one day it was collected on a filter and recrystal- 
lised ; it melted at I90°C. and weighed 0.822 g., corresponding to 0.528 g. 
mannose and or 0.475 g. mannan, and to 31,36^0' of the dry seed. 

The observation of Bertrand mentioned above on the occurrence of 
mannan in coniferous trees led me to look for mannan also in the wood of 
Cryptomcria. I boiled small chips of the wood (dry matter=:9ig.) with 
dilute sulphuric acid of '},% 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 6,35%" mannan. 

The seeds of RJiodea Japoinca were also examined for mannan, since 
starch is absent in them. After removing the shells and pulverising the 
seeds, the treatment was essentially tlie same as above m.entioned. 

The analytical data are as follows : 

Original substance 1 1.2444 g. 

Dry matter S.7i4ig. 

Mannose phenylhydrazon 2.1252 g. 

corresponding to 1.3645 g. mannose. 

Hence this result shows that the dry matter of the seed of RJiodea 
yapotiica zowi^Xn's, 14.28 ^q* mannan. 

While I was occupied with the investigation of the reserve carbohy- 
drates in the seeds of Aiiciiba Japonica, I noticed in a recent number of 
Comptes Rendus de I'Academie des Sciences, November 25, 1901., that 
Mr. Champeiiois 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. Dicbat, who found 
mannan also in the seeds of some Liliaccae. 



On the Genera! Occurrence of Bacillus 
Methylicus in the Soil. 



BY 



T. Katayama. 



A very important function of bacteria in the soil is t!ie oxidation of 
ori^^anic matter, whereby carbonic acid is produced which may partly be 
absorbed by the root and carried to tlie 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, ic is Bacillus uicthvlicus 
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 nietlu'l 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 aerobic 



^ This niiciobo was fust ohscivod Iiy O. Loc.i'. Central Blatt. f. Baktcriologie. 1S9:;, 
* According to the nomenclature of A". B. L<-hviauH this name would have to l>c changed to 
Bacterium methylicum, since it forms no spore?. 



256 T. Katajama: 

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 formate 0-57o 



0/ 
/o 

Dipotassium phosphate 0.2 ,, 

Diammonium phosphate o.i ,, 

Magnesium sulphate O.oi ,, 

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, tliat had 
been suspended in the 10 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. 

i). 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). In stab 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 i fi. thick and 2-3 //. 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. 



On the trcneral Oceiinciuc of Bacillus inetlivHciis in tlic Soil. 



257 



On inoculating' from these wliitc colonies in bouillon and making 
potato-cultures, I observed that this white bacillus resembled in every 
respect, except the color, to th.e red Bacillus vtctJiylicns, 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 tl.e 
Bacillus metliylicus in every instance. 

The results arc seen from the following table ; 



I-oculity. 


Kind of soil. 


I )ate. 

(were inoculated in the 

formate solution.) 


Color of film, 
(after 2 months.) 


This College, Koiiial)a. 


light loam i I* 
(mulberry plantation) ( 2 


14 January 


rctl 

slightly reddish 

(few samples) 

white (many samjile^i 


» » )i 


light loam [ I 
(experimental field, -| 
without manure ( 2 
for 14 years) 


„ 


white 


)j » >i 


light loam ( i 
(Ibrcsl) 1 2t 


„ 


slightly roltlish 
white 


X.xia, 
Sliimosa provincf. 


fertile loam i i 
(firm) ( 2 


iS January 


\\\\\W 


Near the ri\cr Tone, 
Musashi province. 


sandy soil i i 
(rice field) ( 2 


" 


" 


Kuniagai, 
Miisashi pnnlnce. 


fertile, clayey loam 1 i 
(farm) ( 2 


'I 


" 


Kuniagai, 
Musashi province. 


un-manuretl, clayey | i 
loam 
(near rail road) ( 2 


.. 


.. 



* Ihis sample w.is taken from the depth oi i cm. 
t This sample was taken from the depth of 5 cm. 



258 T. Katayaina: On tlie General Ooenrrence of Bacillus nietlijlicns in the Soil. 



Locality. 


Kind of soil. 


Date. 

(were inoculated in the 

formate solution) 


Clor of film, 
(after 2 months) 


Ueda, Shinano province. 


fertile loam. 1 i 
(mulberry plantation) ( 2 


19 January 


wliite 


„ 


un-manured gravelly ( i 

soil ' J 
(mulberry plantation) ( 2 


„ 


» 


Nagano, 
Shinano province. 


fertile, clayey loam. ( i 
(mulberry plantation) ( 2 


" 


j> 


Nagano, 
Shinano province. 


fertile, clayey loam. 1 i 
(forest) ( 2 


" 


)) 


Nagano, 
Shinano province. 


clayey loam. ( i 
(experimental field, -| 
without manuring ( 2 
for many years) 


„ 




Kawasaki, 
Musashi province. 


fertile, sandy loam. ( i 
(orchard) ( 2 


5 February 


„ 


Kyoto, 
Yamashiro province. 


fertile, sandy loam. 
(forest of oak trees) 


1 1 March 


red 



It was noticed very clearly that maiuired fertile .soils yielded also the 
IhicHIks mciJiylicus in a greater r.umbcr than sterile poor soils, to judge 
fron.T the great difference in the time of film developement in the formate 
solution. 

Since this bacillus occurs in j:)utrefying manure, in the air and in every 
one of the examined soils, it can be concluded that it is of general 
occurence. 

I intend to make further investigations in regard to this microbe and 
to its colorless varict)'. 



On the Liquefaction of (Vlannan by IVlicrobes. 

BY 
S. Sawamura. 



Since I liad 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 mucihige in question 
was derived from Hydrangea pauiculata Sieb. var. minor, and consisted 
to a considerable extent of mannan containing also some araban and 
galactan. 'J'he paper Mnanufacturer, 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 t!ie 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 
importar.ce to test various kinds of microbes upon the power of liquefying 
mannan jelly. In Japan occurs in commerce a fo(.d called " A'^//;/m.6/." 
prepared from tl-.e root of Conophallus Konyakn b>- 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 z% of the refined dry product of the s.id root in a 
hot solution containing ^% of pepton and o.,o^, of magnesium sulphate 

^ -l-hls cala.mty u as easily avoids by antiseptic moans, which the writer pa^ixv^xi. 
■^ Tsuji. .\[annan as an .\rtiele of Human Foxt. UuIIctin of this College. \ol. H. No .. 
p. 103. 

Ki„osh,ia. On ti>e Occurrence of Two Ki.uls of M,nnan in the Root of Conophallus Konvaku. 
hid. \ol. II. Xo. 4. p. ,^,3 

V'sid-.iwof,', ilii.l. \ ol. 11. No. 7. p. 4o(>. 



26o 



S. Sawamiira: 



and dipotassiumphospliate. This solution gave on cooling a transparent 
jelly resembling tlx agar jelly, used for bacteriological cultures. 

After the mixture was sterilised in the usual wa}' it was infected with 
various kinds of bacteria and }easts, and kept at 36°C. for two days. 
The results are seen from the following tabic. 



Names of the Microlies infected. 


Liquefaction of Mannan Jelly. 


Sacchaiomyces cerevisia?. 


- 


,, apiculatus. 


- 


„ from the mucilage. 


- 


Micrococus fi'om Koji. 


- 


Streptococcus from sillv-worm. 


- 


Bacillus capsulatus. 


- 


„ cyar.ogenus. 


- 


,, Ilucpp.e. 


- 


„ megatherium. 


- 


„ mesentcricus ruber. 


- 


„ „ \ulgatus. 


+ 


„ pyocyaneus. 


- 


„ prodigiosus. 


- 


,, ty[)lii imiriuiii 




„ Zeiikerci 


- 




sulitiiis. 


- 



On the Liquefaction of Maniian by Microbes. 



261 



It was only Bacillus vieseiitericus vulgixtiis of the sixteen species tested 
that liquefied mannan in two da\'s, 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 .'^evcral 
weeks further, but it was only Bacillus prodigiosiis 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 mcsentei'icus vulgatus. 

Ikit this phenomenon is considerably accelerated when a certain wild 
yeast propagates luxuriantly in the mucilage.^ This influence is difficult 
to exi)Iain, since this yeast in itself has no action on mannan and further 
does not ferment the mmnose formed by the bacterial action. ^ 

I infected sterilised mannan jelly with Bacillus viesciitcricns vulgatus 
alone, and in a second cose with this bacillus and that wild yeast together, 
and in a third case with that bacillus and beer yeast. After keeping the 
jelly at 36*^0. for some days th.e sugar formed was determined with 
FeJiliuz s solution. The results were as lollows : — 



I 



Micro! )cs. 


Len;4li ut 
Culture. 


Strength of 
Solution. 


Sugar found in '^^ 
of mannan. 


Suqilus. 


\'uli,'atus. 


4 days. 


io"o 


, , -C-,0 




Vulgatus + Uic AViKl \'cast. 


" 


" 


i7-675.''o 




\'u!i;atus. 


2 days. 


3,-0 


5742,%' 




\'ulgatiis 4- the Wild \'east. 


" 


>» 


S.614.% 


50 02^ 


A'lilt^atus. 


I day. 




3676"o' 




A^iljjatus + I'ccr Yeast . 






4.0:.; "„' 


3S.o,;%' 



* The accelerating action of yeast on diastase \\as ol)ser\\~<l by .Vt'rn's. CentmUlilalt fiir Agricul- 
turcheniie 1903. p. 286. 

' 0.04 vol. ^0 of alcohol w as formed liy cultivating it in 10% glucose solution at ^6°C for 12 days. 



262 !^« Siiwainura: On the Liquefaction of Mauuau hy Microbes. 

Tlie amount of alcohol formed from man nan 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 r^acilliis nicscntcriciis 
V III gat us. 

Summary. 

While tluis far no microbe was observed liquifying galactan, there 
exists an exception as regards mannan, since Bacilhis ineseuteriais 
vnlgattis can easily liquefy mannan jelly. Bacillus prodigiosiis 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. Kozai who kin.dly provided me with the pure cultures of bacteria 
used in this ex[)eriment, and also to Mr. T. Yaniasakl, Assistant of this 
College. 



Chemical Note on a Singular Phaenogamic Parasite. 



BY 



T. Suda. 



In the province of Tosa and in the southern part of Kiushiu in Japan 
a phaenogamic 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 phrenogam, 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. Tin's mass is of black color. 

This singular plant is a kind of BalaiwpJiora but not identical with 
BalanopJiora 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. 

y. D. Hoocker^ states that Ihilanophora dioica in India has not the 
long branching rhizomes and not so much resin as the related BalanopJiora 
elongata growing in Java. 

TJi. Polcck published some studies on this latter species and found the 
melting point of the resin (Balanophorin) 90-95^ C. 

Several points in regard to the Japanese species of Balanophora 
seemed to mc of some interest, it was the amount of resin present and 
further, the amount of lime and magnesia, since pha:nogamic parasites 
require less lime than green plants. 

The material furnished to mc contained 20.95^0 dry matter and this 
yielded 7.8ij^J of ash. The amount of the resinous compound present, 
extracted by ether, was 15.87^0 of the dry matter. For determination 

» Transactions of the I.innc.in S-xiVty. Vol. XX. T.irt !. (1S63). 



264 T. Snda: Chemical Note ou a Singular Phwuogamic 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 OA2g% 

MgO 0.244% 

This shows that also this phaenogamic paras'te like the well known 
Cusciita 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. 



On the Action of Formaldehyd on Pepsin. 

BY 
S. Sa"wamura. 



I 



Formaldehyd exerts an injurious action on enzyms, as was llrst 
observed by O. Loeiv.^ Pepsin and diastase are killed in one day when 
left in a neutral ^% 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 ^%. At 
low temperature, however, sucrase resists formaldehyd more than other 
enzyms do, as Bokorny^ observed. This author* also stated that a 
formaldehyd solution of \% kills maltase in 24 hours, one of 5^ 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 Pekel- 
flaring'^ who writes : ,, Mit Formal zu einem Gehalt von 2-^% versetzte 
Loesungen von Pepsin in Salzsiiure konnen ohne merklichen Verlust an 
verdauender Wirkung Tage lang aufbewahrt warden." ,, 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." 

* Journ. f. prakt. Chcni. 37, p. 104, (iSSS), 

* Aiin;Jes dc 1' Inskitut Pasteur S, p. 796 (1894). 

* Pfliig. Arch. 85, p. 267. 

* Ibid. 

6 Lot-w ; Report No. 68. U. S. Dept. of Agriculture (1901). 
« Journ. of Expcr. Med. 4, p. 47 (1898). 
' Z. physiol. Chem. 35 p. 29 (1902). 



266 S. Sawanuira: On the Action of Formaldehj d on Pepsin. 

These statements, contradicting apparently the observations of others, 
induced me to make some experiments Avith 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 sohitions were tested as above with the same result. The result of 
my first experinient is more decisive than that of the second, since the 
adhering formaldeliyd 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 PekelJiaring. In the 
first place, my formaldehyd solution was of higher concentration, and in 
the second place, that author tested a solution of pepsin in o 2% 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. 



* Tliis filjiin after heing freshly prepared was kept in glycerol. Before ap[)Iication it was kept for 
some time in very dilute hydrochloric acid and washed. 



Ueber die EInwirkung des //jr^-Brennens. 



vox 



O. Shishido^ Ringakushi. 



Einleitung 

In alter Zeit war das japanischc Inselland vermuthlich vollkommen mit 
Wald bedeckt, und die Bewohner suchten ihren Lebensuntcrhalt im Walde ; 
Friichte miisscn ihnen ganz unentbehrlich gewesen sein. Aber mit dcr 
Zunahme dcr Bcvolkerung kam zuerst die Frage der Erniihrung derselben ; 
man musste also seine Lebensmittel kiinstlich gewinnen, und so cntstand 
unserc Landwirthschaft; daraus folgtc dass die Wald-Bestilnde, welche 
sich in den Ebencn und Gebirgen befanden, mit der Zeit nach und nach 
abnchmcn musstcn ; ja die Bestiinde wurden als ein Hindcrniss 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 Fiillung der Bestande daucrtcn 
bis in spiitere Zeitalter fort. 

Auf diese Weise entstandcn die heutigcn kahlen Gebirgc und bestands- 
loscn ICbencn, welche wir " Ilara " ncnncn. Diese Fliiche an Hara ist 
in Japan ausscrordentlich gross, die folgende Zusamcnstellung gibt uns 
eine ungefilhres Bild davon : (December 33 Meiji. 1900) 

Staatswaldungen 13072602 chd 

Kronwaldungen 2091 7S4 cho 

[Privatwaldungcn, j 

■jGcmeindcwaldungen,^ 743CO91 chC> 

(anticrc WaldungenJ 
Summc '. .... 22594477 chd 



268 0. Slsishido: 

Starts- Ha 7' a 1 434666 c/id 

Yiron-Hai-a i 54174 cho 

{VnvdLt-Hara \ 

1 Gemeinde - //^r^^l 1053462 chd 

■ und andere Hara) 

Summe 264.JJ02 chd 

(i chd = C)()\j. [I. in) 
demnach betriigt die Flachen-Summe der Hara in Japan 2645302 chd. 

In Japan herrscht die Sitte, jiihrlich das Hara-GvdLS, zu brennen ; es 
gehort 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 untersuchtc 
ich die Vegetation, die Boden- und die Wuchs-Verhiiltnisse auf verschiedenen 
Hara. Fiir meinc Arbeit fand ich hinreichenden Stoffin der Gegend der 
Kiyosumi-Schuhvaldungen. 

I. Absehnitt. 

Die GcscJiicJitc der " Kiyosinni-Hara.''' 

Wie erwilhnt, habcn ausgcdehnte Wald-Bestlinde in alten Zeiten das 
ganze Land bedcckt ; diesc Walder wurden aber vielfach abgeholzt oder 
abgebrannt. Solche Zerstorungen des Waldes dauerten bis in die neuesten 
Zeiten herein; Ebene und Gebirge sind solcherart in grossem Massstabe 
in Kahlfliichen {Hara) umgewandelt worden. Die Hara, welche nun auf 
diese Weise entstanden ist, bcnutzt man hauptsachlich zu landwirthschaft- 
hchen Z^Yecken, nilmlich zur Gewinnung von Grasdung, oder gebraucht sie 
als Wicsen ; der Ictzte Fall kommt aber in Japan sehr selten vor. 

Die genante Kiyosumi-7/crrc? ist getheilt in zwei Geimeinde-//<r?n?, d. h. 
jene von Amatsu und von Tojo und einen Theil, welcher zu den 
Schuhvaldungen gehort. Von diesen Gemeinde-//rt;'(i:'i" existiren keine 
urkundlichen Nachweise. Niemand kennt deren Entstehungs-Geschichte ; 
nach der Ansicht der Gemeindevorsteher in Amatsu und Tojo sind diese 
Hara nicht in dcrselbcn Weise entstanden wie die meisten Hara in Japan 



L'eI)or (lie Eiimirkiiiig dcs 7/<f rrr-Brenneiis. 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 iibcrvveisen ; aber in den Provinzen 
Awa und Kazusa (wozu die K'lyosumi-IIara gehort) existirtc diese Sitte 
nicht. 

Diese Gemeinde-//(^y-rt sind in den iiltesten Zeiten noch teihveise mit 
Bestiinden bcdcckt zu denkcn und verblieben der Gemeinde als Freigiiter. 
Man benutzte die Hara hauptsiichlich, um cine Grasart, sogenanntes 
" Kaya " (Miscanthus sinensis Anders) zu gewinnen ; um cin moglichst 
gutes Wachsthum desselben zu erzielen, brannte man diese Hara, jiihrlich. 

Nur iiber die Geschichte der Hara, welche zu den Schuhvaldungen 
gehort, hat man cinigc Kenntniss ; der Wald an Sannodai und derselbe 
an der Nordseite von Suzuriischi, also die jetzige Hara, sind im Jahre 2 
Tempo giinzlich abgeholzt worden, seitdem sind sie ::iuciinal gebrannt 
worden, einmal im 24 Jahre INIeiji (1891) und das zwcitcnmal in r^Ieiji 32 (1899). 

Die Gemeindcvorsteher in Amatsu und Tojo sagtcn mir, dass man 
sich jctzt wiederum cntschlosscn hat zur Aufforstung dieser Hara. 



II. Abschnitt. 

Ziistand der Kiyosniui-Hara. 

I. Kapitel : Boi/t'risus/('i;/ r\-. 
A) La ore und FUiclic. 

Die ¥J\\oiv\\\\\-Hara umfasst drei Hara, namlich jcne im Schuhvald, 
dann jcne von Amatsu, Uchiura, und cndlich die Hara von Tojo, Seijo, 
Hiroba und Ilamaogi ; die ganzc Fliiche der zwei letztercn Gemeinde-//i7;*<7 
hat 391,0325 Jia ; die Fliiche der in den Schuhvaldungen gelegenen //<//v/ 
betriigt ca 20 Jia. 

Diese Hara befindet sich an der Nordostseitc von Awa, angrenzend 
an die Provinz Kazusa ; das Bergland, auf wclchem dicsc Hara liegcn, 
erstreckt sich von dem Platzc der Schuhvaldungen gcgcn das Meer zu. Der 
Lagc nach gchrn-t diese Hara wic Prof. Honda bcstimmtc. der subtropischen 



2/0 0. Shishido: 

Waldzone an. Diesc Hara ist auf Gebirgsauslaufern steiler Ausformung, 
welche bis zum Meere reichen, ausgebreitet unci hat keine ncnnenswerthe 
ebene Fliiche aufzuwcisen. 

B) Bodcn. 

(Grundgestein) 

Dcr Bodcn, aus welchen die Halbinsel Awa gebildet ist, besteht 
hauptsachlich aus drei Gesteinsartcn : Tuff, Schicferthon und Sandstein. 
Diese Gesteinc sind ini Allgemeincn rauh und weich, leicht verwitterbar 
und bilden viele Bodenmodificationen. 

II. Kapitel : Das Kliiiia. 

Das Klima libt natiirlich auf den Pflanzenwuchs cincn grossen 
Einfluss aus. 

I. Die Tcinpcratur. 

Der " Kuroshio " oder warme Meeres-strom, welcher an die Siidost- 
Ktiste der japanischen Insel, von Siidwest nach Nordost bespiillt, iibt auf 
die Lufttemperatur einen grossen Einfluss aus ; namcntlich wirkt er auf das 
Klima der Halbinsel Awa in hochstem 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 Extrcmen. 

2. FeucJitigkcit 

Die Halbinsel Awa hat eine grosse Luft-Feuchtigkeit, wodurch die 
Regenmenge ausserordcntlich anwiichst, im August erreicht die relative 
Luftfcuchtigkcit ihr Maximum und im Fcbruar ihr Minimum. 

3. Dcr Neb el. 

Wo warme und kiihle Stromungen des Meeres sich mischen, wie an der 
Ostseitc der Halbinsel, werden hiiufig dichte Nebel erzeugt ; diese treten 
mcist in den Monaten April bis September auf. 



I 



Uebor die Eiinvfrkuiig dcs //aivf-Brenucits. 271 

4. Frost. 

Er kommt in Kiyosumi schr seltcn vor, Frosttagc sincl in Kiyosumi 
in einem Jahrc nur 20 unci zwar mcist in der Zcit von Ende Oktobcr bis 
Anfang- Mitrz. 

5. Dcr Wind. 

Wind hat auf den Pflanzenwuchs einen grossen Einfluss., deshalb muss 
scin Einfluss beachtct wcrdcn. 

a) HaitptiK^ind. 

Von April bis September (5 Monate) herrscht Westwind, wiihrend in 
den 7 anderen Monaten sich hauptsachlich Nordost odcr Xordwestwind 
cinstellen ; im Allgcmeinen sind Nordwinde die Hauptluftstromungen. 

b) Sturm. 

Es gibt in dieser Gcgcnd oftmals Sturme ; nach den Unlersuchungen 
in Choshi und Fura(32, Meiji, 1899) hcrrschen die ''Jicftigcn Winde" meist in 
Winter, wiihrend in Februar, Miirz, Spiitwinter und Anfangs des Friihjahres 
" 5///;-wr " auftreten. Das Klima ist in Kiyosumi fiir Menschen und 
Pflanzen giinstig zu nennen, da die Temperaturdifferenzen zwischen Sommer 
und Winter sehr gering und die Feuchtigkeitmengcn. welche ztim 
ITlanzenwiichse n(')tig sind, reichlich vorhanden sind. 



III. Abschnitt. 

L iiltitr dcr Gi-oi/ii/ bci Kivosnni:. 

Auf der llalbinsel Awa und Kazusa beflnden sich zahlreiche nicdrigc 
Hohcnziigc, zwischen denen in den Thalcrn cine recht miissigc Flache 
an Ackerboden vorhantlen ist. 

In der Kiyosumi (^icgend, wo das Ilugelland seine grosstc Hohe 
crrcicht, findet man Ackerland nur an der Kiiste in gcringcr Ausdehnung ; 
es ist eben kein Land vorhanden. welches vortheilhaft landwirthschaftlich 



0. Shishido: 



benutzbar wiirc, wenn auch hie und da die Landwirthschaft bis zum 
Berghange sich ausdehnt, wo dessen Gefalle minder steil ist. Die Fliichen 
der Hara des for.st-und landwirthschaftlich benutztcn Boden.s beziffern 
wie folet sich : 



Xamen. 


Ackerflaclic. 


Waklflachc. 


Z^wvfflache. 


Summa. 


Hara 
Procent. 


Kamogawa 


198.31 lia. 


10.84 ha. 


8.36 ha. 


217.51 ha. 




Amatsu 


149-33 ,. 


981.20 „ 


318.19 '. 


1448.72 „ 


21.9 „ 


Kominato 


108.35 " 


751-75 -. 


192.37 ., 


1052.47 ,, 


1S.3 ,, 


To jo 


395-88 ., 


1180.45 " 


228.71 „ 


1S05.04 „ 


12.7 » 


.Seijo 


300.40 „ 


507.40 ., 


102.35 „ 


910.15 „ 


1 1.2 „ 


Kameyania 


544-19 .. 


1152.50 ,. 


391-62 „ 


20S8.31 „ 


1S.7 „ 


Kururi 


275.08 „ 


1527.64 V 


30.40 „ 


1833-12 „ 


1.6 „ 


Omi 


140.15 „ 


247.91 -, 


69.4S „ 


457-55 M 


I5-I » 


Oikawa 


263.46 „ 


1756.36 „ 


317-45 ., 


2336-27 » 


13-5 ^> 


Xischiliata 


657.92 „ 


904-99 - 


1992.01 ,, 


3554.92 „ 


56.0 „ 


Otak; 


333-83 „ 


313-70 „ 


312.10 ., 


959-63 ^, 


32-5 ,, 


Fusano 


817-71 ,- 


103355 -' 


476.S4 -, 


2328.10 „ 


20.5 „ 


Fusamoto 


447-81 V 


291.34 „ 


192.46 „ 


931-61 „ 


20.7 ,. 


Katsuura 


204.53 •> 


191-33 -, 


159-90 „ 


555-71 „ 


28.7 „ 


Seikai 


240.16 „ 


536.7S ,. 


140.03 „ 


922.97 „ 


15-S „ 


Ueno 


569.34 .. 


769-59 - 


49.00 „ 


1387.93 =, 


3-6 „ 



Man sieht, dass der Waldgrund im Allgcmeincn die Ackerkulturfluche an 
Grosse iibcrwiegt, namenthch hat Amatsu einc Waldfliiche von 6y,y%y 
daher ist auch die Hauptbeschaftigung der Lcutc dortselbst die 
Verkohhmg des Holzes, Holzschlag, Ilolztransport u. s. w. ferner bemerkt 
man die grosse Ausdehnung der Ilara ; diese Ilara wcrdcn zwar thcils zur 
Futtcr- und Kayagewinnung bcnutzt, aber thatsiichlich geben die " incistcn'" 
Thcile der Ilara in diesen Gegenden nahczu keinen Ertrag. 



I'ober (lie Eiinvirkiin^ dos //rt rre-Bronnens. 



-/3 



IV. Abschnif. 

])ic Ilara unci iJirc Bcsitr^vcrhiiltnissc. 

I. Kapitel : Bcsitzarten : 

(A) Dcr Univa-sitixt gcJidrcndc Hara. 

Ein Thcil dcrselbcn befindet sich in Kiyosumi und andcrer liegt in 
KiwaJa, an dcr Nordseitc der Schulwaldungen ; die ganze Fliiche der 
Univ. Hara beziffcrt ca 20 ha ; sie soil jedoch baldigst in Wald 
umgcwandelt werdcn. Unter diescn Hara gibt cs cincn Theil, die Hara 
Musadogadai, dcrcn Benutzungsrecht den Leuten von Kiyosumi verguns- 
tigungsweise zur Gewinnung der Kaya (:\Iiscanthus sinensis Anders) 
iibcrlasscn wird, dafiir miissen sic dann der Universitilt gewisse 
Gegenlcistungcn stcllen. 

(B) Die Geincindc-Hara. 

Den Haupt-Thcil der sogenanntcn Kiyosumi- //^//-^znimmt die Gcinciudc- 
Hara ein, von dcr zwei Arten unterschieden werden : crstciis die 
Hara von Amatsu und Uchiura deren totale FUiche 31S ha. betragt, 
zwcitcns jcnc von Tojo, Scijo. Hiroba und Hamaogi mit eincr Fliiche von 
8S.9 ha. Die crstcrc bildct die sogenanntc Mukomine-//<?/v? welche nordlich 
an die Schuhvaldungcn grenzt. und sich nach den Kiistcn Amatsu und 
Uchiura crstrcckt ; die zweite llara reicht von den Schuhvaldungcn "-eo-en 
Toju liin. 

11. KAPrrKL: (ii-7i'inriiini^ dcr Haraf^roduktc. 

Auf tier (itiiicidc-Ifara kiMincn die Bcrcchtigten nach ihren Bedurfnissen. 
den Anwuchs dcr Ifara crntcn. es gibt kcincrlci Beschninkung darin. 
sei cs class Jcmaiul scincn Frntcantheil an Gras verkaufen odcr cinem 
Anilcni unentgeltlich iibcrlasscn will ; nur die Gcwinnungszeit der 
Produkte ist ctwas bcschrilnkt ; cs muss namlich die Gewinnung der Kava 
im December, uml jcnc der andcrcn Graser, von Juli bis August statt- 
tinilcn ; in dcr dcr l^niversit;it gcluircmlcn ffara ist die Gewinnuncr dieser 



2/4 0. Shishido: 

Griiser principiell nicJit gestattet, nur in Musadogadai, welches ebenfalls 
zu den Schuhvaldungcn gehort, konnen die Leute in Kiyosumi, nach 
Erforderniss Kaya bekommen, gegen gewisse Gegenleistungen. 

V. Abschnitt. 

Das Brenncn dcr Hara. 

I. Kapitel : Zii'cck des Brcmiens. 
(A.) Entstchitng aus alter Sittc. 

Die Entstehung der Hara ist im Allgemeinen dem Nicderbrennen des 
Waldes zuzuschreiben, man woUte diese Flache eben nicht wieder Wald 
werden lassen und branntc sic deshalb von Jahr zu Jahr, um Gras als 
Futter und Diingungsmittel zu bekommen, namentlich war Kaya das Haupt- 
produkt. Die Sitte des Harahxcwwaw-, dauert bis Heutzutage, so werden 
z. B. die meisten Theile der YAyoswmx-Hara noch jetzt jiihrlich gebrannt. 

Der Vorsteher in Amatsu sagte mir, dass die Leute in diesen Gegenden 
der Ansicht huldigen, dass die Pflanzen der Hara durch das Brennen 
kriiftiger Averden. 

(B.) LaiukvirtJiscliaftlichc Boiutzung dcr Hara. 

Da wie erwillint in diessen Gegenden nur gering Menge an Ackerflilche 
sich vorfindet, so bedarf cs hier wenigen Grases als Diingmittel ; audi 
erhalten die Leute grosse Mengen Fischdiinger, welcher fur den Acker- 
boden wirksamer ist. 

(C.) Wcrth des Harabrennens r.ur Gczi'innung 
von Futtergriisern. 

In Kiyosumi u. Umgegcnd wird eine grosse Zahl Kinder gehalten 
welche fiir den Transport des Holzes nicht entbehrt werden konnen ; man 
bedarf fiir den Untcrhalt dieses Viehcs vicl Futtergras, welches allein 
auf dcr Hara gewonncn wird. Um nun Grilser fiir Futterzwecke in 
moglichst grosser Mcnge zu erhalten, brennen die Baucrn die Hara 
alljahrlicli. 



Ueber die Einwirknng des //aiYr-Brennens. 275 

II. KAriTKi. : Verfahrcn dcs Harabrenncns. 

Wcr HaragXTX.'s, brcnncn will, hat dies bei dcm Gemcindevorsteher 
anzumcldcn, wclcher dann Nachricht davon an die Polizeibeh5rde 
gibt. Wenn die Erlaubniss ertheilt ist, beginnt man mit dem Brennen des 
Haragv^ss,ts, nachdem die Absicht u. den Zeitpunkt dcs Brennens den 
benachbarten Bodeneigenthiimern mitgetheilt wurde. 

III. K.M'ITEI. : Zdt des Harabrenncns. 

Die Ernte des Kaya (Miscanthus sinensis) erfolgt in jener Jahreszeit, wo 
das Kayagras seine maximale Hohe erreicht hat, das ist in der Recrel 
Mittc December der Fall; die iibrig bicibcndcn Griiser auf der Hara 
brennt man dann im Monate Februar ; bald nacli dem Brennen begriint 
sich die Hara wieder. und schon Ende April ist die Hara mit neuem 
Graskleide bedeckt. 

I\\ KAi'iTF.r. : Die Methode des Harabrcnnens. 

Sobald die rolizeibehorde dem Eigenthumer eincr Hara die Erlaubniss 
zum Harahx^nncn gegeben hat, beginnt derselbe mit dem Brennen der 
Hara an cinem stillen Tage, wobei ein Polizei- und ein Gemeinde-Beamter 
sowie der benachbarte Bodeneigenthiimer gegenwiirtig scin sollen. Auf 
den Grenzlinien stellen sich Leuten an um benachbarte Hara odcr Wald 
vor dem iiberlaufenden Feucr schutzen zu konncn. 

VI. Abschnitt. 

Phrsikalise/ie Eigenschaften des Bodens. 

I. KaI'ITKI. : /// Kiyosunii. 

A) Tiefe. 

Die Tiefe des vcrwitterten Bodens ist nach C^rtsvcrhaltnissen 
verschiedcn. namlich in den ThlUcrn ist er tiefer als auf den Bergen, wctl 
der Boden am Abhange nach und nach thalwarts herabgcfiihrt wird durch 
Regen unti andere atmosphiirische Niederschliigc. 

Die Tiefe der \'erwitterungsschicht wdche ich auf verschicdencn 
Probeflachcn untersucht habe ist folo-cndc : 



2/6 



0. Slnshi(l(K 



Xaiiien dor Proljcfiachc. 


FJiicbe. 


Xe;gung 


Staiidoit. 


ilurchschnittliche Tiefe. 


Kiridoscbi (Xord'cite) 


20 rj. m. 


35° 


am Miltelliang 


0.29 m. 


„ (Siidseitc) 






•' 


0-35 " 


Kajisaka (Nordseite) 


, 


39° 




0.41 „ 


(,Sadseite) 




32° 


" 


0.28 „ 


Omiyama (Xordseite) 




34° 




9-27 V 


I. (Siidscite) 




34° 


V 


0.50 „ 


,, 2. (Siidseitc) 




36° 




0.47 ,. 


S.innodai (Xordseite) 




35° 




0.21 „ 


„ (Siidseite) 




31° 




0.61 ,, 


Suzuriischi (Xordseite) 




32° 


M 


0-33 ', 


„ (Siidseite) 




•-)-.0 




0.29 „ 


Musadogadai (Xordseite) 




32° 




O.S3 <' 


(Siidseite) 




29° 




0.44 V 


Xanamagari (Xordseite) 




35° 




0.45 >' 


C! (Xordseite) 
Sengen . ^ 

^ (Siidseite) 


>, 


35° 
34° 




0.S2 „ 
0.71 ,» 






dur 


chsclmittllch. 


mitteltiefgriindig 0.434 m. 



Immer wird man auf oft gebrannter Hara eiiien scichtcn Bodcn finden, 
wiihrcnd der seltcn gcbranntc Nir rahoden oder Waldboden viittclticf- bis 
ticfgrilndig ist. 

Auf Grund meincr Untersuchungen komme ich zai dem Schlusse dass 
dcr HarahodQW in Kiyosumi im Allgcmeincn mittelticf-griindig ist. 

T)) Biiidigkcit. 



Die 15indigkcit dcs Bodens ist in den cinzelnem Theilen der Hara etwas 
verschicden, je nachdem gewisse Parthien der Ilara jalirlicJi oder pcriodiscJi 
gebrannt wurden ; die jahrlich gebrannten Theile haben lockcrcn oder grab 
gekriimelten Boden, wahrend auf selten gebrannter Hara die Struktur 
feiner und gekriimclter ist. 



Uobor (lie Eiinvirkuiig des //irfrrt-Breniieiis. 277 

Im Allgemcincn abcr kann man den Bodcn auf der ¥iiyosum\-I/ara 
wegcn dcr Luftfcuchtigkcit als zicmlicli mild odcr locker ansprcchcn. 

C) FeucJitigkcit. 

Die Harcx, deren Bodeniiberzug jahrlich gebrannt wird, mlisstc cigcntlich 
starker ausgetrocknet sein, weil der Bodcn oftcr und liinger direkt der Luft 
ausgcsetzt ist ; aber or ist dennoch durchwcg ziemlich feucht, der Grund 
davon ist ebcnfalls darin zu suchen, dass die Luftstromungcn auf diescr 
Halbinsel an sich sehr feucht sind, so dass disscr Unterschied nicbt so 
fuhlbar wird. 

II. Kapitei,: Ausscrc Ziistandc der Kiyosv.mi-Hara. 

Die Hara in Kiyosumi wurdc friihe schon von Wald-Bcstiinden entblosst, 
jetzt ist die ganze Fliiche dersclben hauptsachlich von Kaya Grass 
(Miscanthus sinensis Anders) bcdeckt. Abcr die ]\Icngc dcr Kaya ist 
nattirlich je nach dcm Wicderholungszcitraume des Brennens verschieden ; 
in alien jencn Thcilen, wclche zvenigcr oft gebrannt wcrdcn, mcngcn sich 
Baumarten (wie hauptsachlich Oucrcus glandulifera, Ouercus serrata, 
Ouercus acuta, Ouercus vibraycana, Machilis japonica, Evonymus curopeus 
var. hamiltonianus, Rhododendron indicum var. Kaempfcri, Salix Sicboldiana. 
Spiraea callosa n. s. u.) und Halbbiiume den Miscanthusartcn bci. Aus 
dicscm Umstandc kcinncn wir Icicht crkcnnen dass die Hara, wcnn man 
sie fill- langere Jahre in Ruhc liessc, wicdcr zum ihrem eigcntlichcn 
Urzustande kommen, d. h. in Wald sich umwandcln wiirdc. So kann man 
z. \\. in Nanamagari, wo die Hara seit 10 Jahrcn )iiiJit gebrannt wurdc. 
folgcndc liaum- und Ilalbbaumartcn beobachtcn : 

Ouercus glandulifera 

Oucrcus serrata 

ICvonymus alatus \"ar. subtriflora 

Spiraea callosa 

luu-ya Japonica 

Dicr villa glanduliflora 

X'iburnum dilatatum 



2/8 0. Shishido: 

Machilus Thunbergii 
Clethra barbinervis 
Smilax china 
Ouercus acuta 
Oucrcus myrsinaefolia 
Rosa multiuora 

Rhododendron indicum \'ar. Knempferi 
Rubus palmatus 
Litsea hypoleuca 
SaHx Sieboldiana 
etc. 
Auf " alljaJirUcJi " gebranntener Hara konncn nur Grasarten vorkommen, 
welchc ich in einem spiiteren Abschnitte besonders anfiihren werde. 

III. Katitel : Das Vcrhaltniss ziviscJien Bodcn iind Kliina. 

iyerivitteriings process) 

Der Boden ist das V^erwitterungsprodukt der Grundgesteine, und 
die Verwitterungsgrosse hiingt von dem KHma ab, welches auf dem 
betrachteten Platzc herrscht. Die Verwitterungsschicht bedeckt den Boden 
in den Gebirgen bei Kiyosumi in sehr verschiedener Dicke. Da der 
Boden der Kiyosumi Hara aus den Gesteinen der Tertiiirperiode gebildet 
ist, so sollte die Verwitterungsschicht eigentlich ziemlich bedeutend sein, 
aber die Wirkhchkeit zeie^t uns eine ^rrosse Unfjieichkeit des //(7/-(7bodens. 
Er ist in seiner Verwitterungsschichte verschiedentlich schr flacJigrilndig, 
namenthch ist dies am Kamme und in oberen Hange zu bemerken ; der 
Grund hiezu hcgt wohl in der Steilheit der Gebirge, wodurch bei 
Regengiissen die verwitterten Bodentheilchen, welche keine geniigend 
schiitzende Pflanzendeckc haben, abgeschwemmt werden. Diese Erschein- 
ung muss natiirhch auf der Hara intcnsiver als im Walde sein, weil der 
Haraho(\<i\\ dirckt frci der Atmosphi'irc ausgesetzt ist, wilhrcnd im Walde 
der ganzc l^odcn durch die Kronen der Bilumc eine Uberschirmung 
erfiihrt. Die Verwitterungsschichtcndicke wurdc in einzelnen HaraXhcxXo.n 
folfTcndermasscn cfefunden : 



Uober dio Eiinvirkiiiif? des //aJVc-Breiinfiis. 



279 



Xamen. 


Jahrc des //arahrennens. 


Dicke der Ver- 
witterungsschicht 


Mind. 


Kiridoshi 


(Nord seite) 
(Slid seite) 


alljalirlich tjcbrannt 


69.6 ni. m. 
73-1 „ '. 


■ 71-3 ni- 


m. 


Kajisaka 


(Xord seite) 
(Slid seite) 


)5 >> 


52.3 V 
67.7 » » 


• 60.0 „ 


•' 


(Jniiyaiiia 


(Xord scilc) 
(Slid seite) 


fiir (Irei Jalire niclit gebraiiiit 
fiir 3 Jahre nicht gebrannt 


84.2 „ „ 
95-3 " „ 


I S9.S .. 




Musadogada 


(Slid seite) 


fiir 6 Jalirc niclit gebrannt 


1 13-3 » ,' 


1 13.3 .. 


- 


Omiyama 


(Sud se:te) 


" 


72.1 .. „ 


72.1 .. 




Suzuriisclii 


(Slid seite) 


iiber 8 Jahre niclit gebrannt 


100.5 " -' 


100.5 • 




Sannodai 


(X'ord seite) 
(Sud seite) 


bis 24 Meiji alljalirlich und spiiter 
im Jahre 33 Meiji gebrannt 

gleich 


66.9 „ „ 
1338 ,. „ 


) 

j- I00.4 .. 




Suzuriisclii 


(X'ord seite) 


„ 


121. 2 . ., 


121. 2 „ 


« 


Xanamai^ari 


(Xord seite) 


fiir 10 Jahre nicht gebrannt 


1 10.7 „ ., 


1 10.7 .. 


„ 


Musadofj;ada 


(Xord seite) 


iiber 10 Jahre nicht gebrannt 


9S.1 „ „ 


98. 1 ., 


,■ 


Sen gen 


(Xord seite) 
(Slid seite) 


Urwald 


S8.4 „ „ 
97-2 ., „ 


92.S „ 


•• 



(D'e Xeigung der einzelnen //iir.r ist in der vorgcgangenen Tabelle bcreits bezeichnct worden.) 

Die Vcrwittcrungsschichten u. tlamit der produktive Boden werdcn 
auf der ungcschiitztcn Hara nach und nacli immer scichter durch die 
Abschwcmmung der feineii Theile thalwarts. Die Xiitzlichkeit u. Xot- 
wendigkeit der X'erwitteruiig.sschiclit fiir tlen Ptlanzenwuchs brauchc ich 
kaiim weiter zii besprcchen. 



VII. Abschnitt. 



Vcrgh'icJi der Effcktc des Brcnnois der Ilara in 
vcrsc/iicdcnon Brcnn- Tcnniujts. 

Uni den Kintluss ties brennens Hara besser zu bcobachten ist nothig. 
dass man ausfiihrliche Untersuchungen auf vcrschicdenen ffara's macht. 
Die nieisten Theile tier Cicnieinde-Z/i?/-,'/ in Kix'osunii siiul. wie ich schon 



2h'0 



0. Slii.^^hido: 



erwahnte, aUjiiJirUcJi gcbrannt, wuhrend die der Schuhvaldung gehorende 
Hara mxr sehr selten gebrannt wurde ; die Ilai-a in Kiwada, welches audi 
der Universitiit gehcirt, wird ebenso alljahrlich gebrannt. Ich untersuchtc 
nun folgende Theile : 



1. Kiridoshi und Kajisaka. 

2. Omiyama. 

3. Omiyama (Siidseitc) und 
Musadogadai (Siidseite). 

4. Suzuriischi (Siidseite). 

5. Sannodai und Suzuriischi 
(Nordseite). 

6. Nanamagari. 

7. Musadogadai (Nordseite). 



alljilhrhch gebrannt. 

seit 3 Jahrc nicht gebrannt. 

seit 6 Jahrc nicht gebrannt. 

iiber 8 Jahre nicht gebrannt. 

bis 24 Meiji (189 1 ) alljahrlich und spiltcr 

im Jahre 33 Meiji (1900) gebrannt. 

seit 10 Jahren nicht gebrannt. 

liber 10 Jahre nicht gebrannt. 



ausserdem habe ich den Urwald SeuQ-cu zum Versrlcich herangezogen. 



I. Kapitel : Prilfiing viittcls Probefliiclicn. 

Um den Antheil der vcrschiedenen Grasarten an jeder Hara 
festzustellcn, habe ich Probeflachen von 2 D. m. genommen, und gebe nun 
die Mittel aus jc drei Proben. 

Kiridoshi 1 Nordseite). 

(alljahrlich gebrannt) 
(Neigung=:35=) 
(Datum 10/4 u. 25/4) 



lateinischcr Name. 


japan ischcr Name. 


Zahl der 

Grasei- 

auf2i_J.n1. 


ox 
/o 


Miscanthus sinensis Anders. 


Susuki 


318 


38.6 


Sanguisorba officinalis I.. 


Waremoko 


80 


9-7 


Senecio kranicri Fr. ct Sav. 


Yaburegasa 


74 


9.0 


Thalictruni minus L. var. elatum Lecoy. 


Akikaramatsu 


67 


S.I 


Arundinclla am^mala Slcud. 


Todashiba 


51 


6.3 


r'.cctlantlius <j;laucocalyx Max. vai". jai--onicus Max. 


Hikiokoslii 


37 


4-7 



r 



Ueber die J^iinvirkuug des Jfard-Broumns. 



281 



Lespcdeza bicolor Turcz. 

Saussurea Tanakae Fr. et Sar. var. pliyllolepis Max. 

I'teris aquilina L. 

Potcntilla fragarioidcs I., var. tcrnata Max. 

Lysimachia cletliroides Duby. 

Viola silvestris Kit. var. gryposera? A. Gr. 

SeiTatula atriplicifolia B. et II. 

Dioscorea Tokoro Makino. 

Polygonum cuspidatum S. ct Z. 

Cirsium japonicum DC. 

Atractylodcs lyrata S. et Z. 

Carex breviculmis R. Br. 

Poa trivialis L. 

Seseli libanostis Koch. var. daucifolia DC. 

Cirsium spicatum (Max). 

Euphorbia Esula L. 

Mancttia ignita (Veil). 

Ilcteropappus hispidus Less. var. isochaetus Fr. et Sav. 

Viola japonica Langsd. 

Allium japonicum Rgl. 

Aster indicus L. 

Artemisia vulgaris L. var. iixlica Max. 

Disporum sessile Don. 



[Ilagi] 

Toliircn 

Warabi 

Mitsubatsucliiguri 

Okatoranoo 

Tach i tsubosu ni i re 

Kumatoribokuchi 

Onidokoro 

Itadori 

Xoazami 

Okera 

Ao;u.i;e 

Ilimeichigotsunagi 

Ibukibofu 

Vamaazami 

Ilagikuso 

Kwaenso 

Vamajinogiku 

Ko sumii-e 

^'ama rakl<yo 

Vomcna 

Vomogi 

Ilr>chaku?d 



34 
26 
16 
16 
15 
13 
12 
II 
10 
S 
6 



41 
3-2 
1-9 
1.9 
1.8 

1-5 
14 
1-3 
1.2 



54 



S::; 



Kiridoshi (Siidseitc;. 

(alljiihrlich gebrannt) 
(Neigung = 33^) 
(Datum 10/4 u. 25/4") 



lateinischcr Name. 



Miscanthus sinensis .\iu!crs. 

Imierata aruiulinacea Cyr. var. KoeniL;ii (Penth). 



japani^cher Xamc. 



Susuki 
Chigava 



/-,ihl der 



Grascr 

in 2 i"!. in. 



344 
»73 



31.6 
N.9 



282 O. 

Brachypodiuni silvaticuni R. ct S. 

Inula salicina L. 

Miscanthus saccliariflorus Hack. 

Sanguisorba officinalis L. 

Lysimachia clethroides Duby. 

Viola Patrinii DC. var. chinensis Ging, 

Serratula ati-iplitifolia 15. et H. 

Pteris aquilina I-. 

Smilax china L. 

Thalictrum minus L. var. datum l.ecoy. 

Athyrium nipponicum Bak. 

Senecio Krameri Fr. et Sav. 

Plectranthus inconspicuus ]Miq. 

Euphorbia Siebokliana Mor. et Dec 

l.espedeza Siebokli Miq. 

Cirsium spicatum Max. 

Aster indicus L. 

I'olygara japonica Houtt. 

Patrinia scabiosaefoh"a Link. 

Dioscorea Tokoro Makino. 

Potentilla fragarioides L. 

Polygonum cuspidatum S. et Z. 

Vitis Thunbergii S. et Z. 

Seseli Libanostis Koch. var. daucifo'.ia DC. 

Gymnadenia conopca K. Pr. 



Sliishido : 








Yama kamojigusa 


140 


12. 1 




Kasenso 


64 


5-5 




Ogi 


62 


5-4 




Warcmoko 


54 


4.7 




Okatoranoo 


40 


3-5 




Sumire 


36 


31 




Ku matori bokuch i 


35 


30 




Warabi 


33 


2.8 




[Sankirai] 


25 


2.1 




Akikaramatsu 


24 


2.0 




Inuwarabi 


18 


1.6 




Yaburegasa 


16 


1.4 




Yamahakka 


16 


1-4 




Xatsutodai 


15 


1-3 




[Hagi] 


8 


^ 




Yama azami 


7 






Yomena 


6 






[Hime Hagi] 


6 






Ominaeschi 


6 






Onidokoro 
Kijimuschii-o 


5 
5 


> 3-6 




Itadori 


5 






Ebizuru 


4 






Ibukibofu 


4 






Chidoriso 


1 





"57 



Aus den Tabelien, ersicht man " dass auf jahrlich gebramitcr Hara'' 
hauptsilchlich die folgende Grasarten auftreten : 
Miscanthus sinensis Anders. 
Sanguisorba officinales L. 
Serratula atriplicifolia B. et H. 
Senecio Krameri Fr. et Sav. 
Ftcris aquilina L. 



Ucbor (lie Kiinvirkimy: dcs //«i'a-Br«'aii('iis. 



283 



Thalictrum minus L. var. datum Lcroy. 

Polygonum cuspidatum S. et Z. 

Impcrata arundinacca var. Kocnigii (Kcnth.) 

l^otentilla fragarioides var. tcrnata Maxim. 

Brachypodium siivaticum R. et S. 
etc. 
Dicse Grasarten sind vorzugsweise Lid it gr User, d. h. sie konnen moist 
nur auf nacktem Bodcn gedeihen, oder kommen wenigstens ofter u. vorzugs- 
weise in nahrungsarmem Boden vor. Auf der Hara, welche scit (^//v/ Jahren 
iiicht gcbrannt ist, sind dagegen die folgcnde Grasarten anzutreffen 
gewesen. 



Omiyama (Xordseite). 

(fiir 3 Jahrc nicht gebrannt). 
(Xeigung = 34-). 
(Datum 1 14 u. 26/4). 







Zahl der 




laieinisclicr Xanic. 


japauisclier Name. 


Gr. in 

2 Q m. 




Miscanthus sinensis Anders. 


Susuki 


295 


--l-^~% 


Smilacina japonica A. (jr. 


Vukizasa 


Si 


6.9 


Aster japonicus Mi(|. 


Vam;ishirogiku 


65 


5.6 


Care.x breviculmis R. Vs . 


Aosuge 


53 


4-5 


Artenn'sia vulgaris L. var. imlica Max. 


Vomogi 


41 


3-5 


Potentilla iVagarioides L. var. tcrnata Max. 


Mitsubatsuchiguri 


39 


3-3 


Cypripediluni ja|)onicuin 'liiunh. 


Kuniagaiso 


37 


3-^ 


riectrantiuis inconspicuus Mit|. 


\anuhakka 


34 


-•9 


Cirsium japonicuni DC. 


Nai^ami 


3^ 


-•7 


Pctasitcs japonicus Mi(|. 


Kuki 


31 


2.0 


Carcx conica IJotitt. 


IIinK'ka!)^u^;e 


-5 


2.2 


Viola Patrinii DC. var. chinen>is Ciinj:. 


Sum ire 


20 


1.7 


Kraunhia florihunda (Willd) Tauh. 


1-uji 


iS 


>-5 


Viola silvcstris Kit. var. grypoceras .\. Clr. 


Tachilsubosumiiv 


17 


1-4 


Polygonum cuspidatum S, oi /. 


Itadori 


17 


14 



] 



2 84 



0. Shishido 



Lysimachia clethroidcs Duby. 

Euphorbia sieboldiana Mor. ct Dec. 

'riialiclruni minus L. var. datum Lecoy. 

Crcpis jai onica Benth. 

Dioscorea Tokoro Makino. 

I'oa trivkilis L. 

I.actuca squaiToba JNIicj. forma iiidivesa Max. 

I'olygara japonica Houtt. 

Erigeron annuus Pers. 

Aspklium dissectum Mett. 

Disporum sessile Don. 

Disporum pullum Salist. 

Cirsium spicatum Maxim. 

Clematis recta L. var. paniculata Tliunb. 

Carpesium abrotanoides E. 

Eupatorium japonicum Thunl). 

Acantliopanax pivavicatum S. ct Z. 

ArundincUa anomala Steud. 

Gentiana scabra Bge. var Buerger! Max. 

Scnecio krameri Fr. et Sav. 

Pteris atjuilina L. 

Sanguisorba eflicinalis L. 

Plectranthus graucocalyx Max. var japonicus Max. 

Saussurea Tanakae Fv. et. Sav. var. phyllolepis Max. 

Serratuki atriplicifolia B. et LI. 

Manettia iginta (^'e^.) 

Atracty lodes lyrata S. et Z. 

Aster indicus E. 

Patrinia villosa Juss. 

Brachypodium silvaticum R. et .*>. 

Osmunda rcgalis E. var japonica Milde. 



Okatoranoo 

Natsutodai 

AkikaramatbU 

Onitabirako 

Onidokoro 

Flimc icliigotsunagi 

Honba akinonogcschi 

[Himehagi] 

Himejoon 

I loshida 

I lochakuso 

Tochikuran 

Yamaazami 

Senninsu 

Yabutabako 

Saua hiyodori 

Ukogi 

Todashiba 

Rindu 

Yaburegasa 

Warabi 

Waremoko 

Ilikiokoslii 

Tohiren 

Kumotorihokuclii 

Kwaensu 

Okera 

Yomena 

Otokoeshi 

Yamakamoji gusa 

Zenmai 



i6 
i6 
i6 
i6 
IS 
15 
15 
15 
15 
15 
14 
14 
14 
13 
13 
12 
12 
12 
12 
12 
12 
12 
10 
8 
8 
8 

7 
6 

5 



1 170 



1-4 
1-4 
1.4 
1.4 
1.4 
1-3 
1-3 
1-3 
1-3 
1-3 
1.3 
1.2 
1.2 
I.I 
I.I 
i.o 
i.o 
1.0 
1.0 
I.o 
I.o 
I.o 

0.9 



V 42 



Uel)or (lie Eiinvirkims: Acs //rtJYt-Brenneiis. 



28: 



Omiyama (Siidscite). 

(fiir 3 Jahrc nicht gcbrannt; 
(Neigung = 34-) 
(Datum T 1/4 u. 26/4) 







Zahl dcr 




latoinischer Name. 


japanischer Name. 


Gr. in 

2 Q. m. 


- 


Miscanthus sinensis Anders. 


Susuki 


271 


22.4 


Clirysantlicmum sincnsc var. japanicuni Max. 


Ryunugiku 


75 


6.2 


Disporum sesile Don, 


Ilochakuso 


52 


4.3 


Carcx lircviculmis R. I'.r. 


Aosuge 


51 


4-2 


As^jrimonia visciduia ISgc. var. japonica Miq. 


Kinmizuhiki 


39 


3-2 


lloutluynia cordata Tluinh. 


Dokudami 


37 


3-1 


I'Icclranllius inconspicus Micp 


Vamahakka 


35 


2.9 


('irsiuin spicalum .Maxim. 


^'amaazami 


31 


2.6 


Tlialictrum niiiuis I., var. datum I.ccoy. 


Akikaramatsu 


30 


2-5 


Senccio Kranxri ¥v. ct Sav. 


Vaburcgasa 


29 


2.4 


Artemisia vulgaris 1-. var. indica Max. 


Vomogi 


29 


2.4 


.\tliyrium nipponicum Ilal^. 


Inuwarabi 


26 


2.1 


Carex lanccolata lioo'.t. 


I likx^uge 


25 


2.1 


(Ixalis corniculala T . 


Katabami 


25 


2.1 


Poa trivialis L. 


Ilinie ichigotsunagi 


25 


2.1 


Dioscoria Tokoro Makino. 


Onidokoro 


22 


I.S 


Cientiana scahra Iv-je. var. Uuergeri Max. 


Kindu 


21 


1-7 


Pacderia tomenlosa I'.l. 


Ilekusokazura 


21 


1-7 


Andropogon micrantluis Klli. 


I lime aburasusuki 


20 


1.6 


Dianthus superbus I.. 


Kawaranadcshiko 


20 


1.6 


Crcmatis recta L. var. peniciilata Tluinl^. 


Sonn'nsu 


20 


1.6 


Astiliie thunlicrgii Mi([. 


Toriashishoma 


20 


1.6 


Crepis japonica Bentli. 


Onitabirako 


20 


1.6 


Lysimaciiia clethixiidcs Duhy. 


Oltatoranoo 


l^ 


1.5 


I.ygodium japoniciim Sw. 


Tsurushinolju 


i: 


14 


l.actuca ihunliergiana (.\. (h.) M.ixim, 


Xigana 


«7 


1.4 



286 



0. Shisliido: 



Aster jnponicu> Miq. 


Yamashirogiku 


17 


1-4 


Aster Fcaber Thunl). 


Shirayamagiku 


i6 


1-3 


I'teris aquilina L. 


Warabi 


i6 


'•3 


\'itis thunbcrgii S. ct Z. 


Ebiziiru 


15 


1-3 


Tricyrtis japon'ca M\'\. 


Ilototogisuso 


15 


1-3 


Rubus incisus Tliunl). 


[Xigaicliigo] 


15 


1.2 


I.cspcdcza pilosa S. ct /. 


Nckoliagi 


15 


1.2 


Brachypodium silvaticur.i R. it S. 


Yamakaniojigiisa 


15 


1.2 


Potentilla fragarioidcs L. 


Kizimushiro 


15 


1.2 


Smilacina japonica A. C'.r. 


Yukiznsa 


14 


1.2 


Carpesium alirotanoidc? 1.. 


"S'abutabako 


14 


1.2 


Sanguisorba officinalis I . 


Warcmoku 


13 


i.i 


Atractylodcs lyrata S. ct Z. 


Okera 


II 


0.9 


Patrinia scabiosaefolia I.inlc 


Ominacscl) i 


8 


\ 


Carex conica Boott. 


I lime kan?ugc 


8 




Kupatorium japonicum Thunl). 


Saw a hiyodori 


6 


2.1 


Saupsurca Tannkac Fr. ct Sav. var. pliyllolcpis Max. 


Tu liircn 


3 


, 



Es wird daraus klar, dass dicse Hara (Omiyama) verhiiltnissmassig- vide 
Grasspccies cnthalt ; cs .sind hier die crwiihntcn Lichtgrilscr seltcncr, 
wahrcnd anderc Artcn in grcissercr Zahl gefundcn wcrdcn. Aus dem 
Aufwiichsc kann die l^odcnbcschaffenheit beurthcilt wcrden ; es gedeihen 
auf scJilcclitcm Eoden cben schlcchtc Grasartcn und auf j^utcni ]?odeii die 
bcssercn Arten. 

Audi die grosserc Zalil der Gcwi'ichsartcn deutet an, dass ein Boden 
nahrungsrcicher ist ; in nahrungsarmem Boden konncn nur wenigc Species 
fortkommcn bci sonst glcichcn A'erhidtnisscn. ]\Ian kann daher schliessen. 
dass die rjucrst genanntc ITnra (Kiridoshi 1 nalirimgsiirmcr ist als die zzvcitc 
(Omiyama). 

II. KAriri;!,. lu^lgcn dcs /yraiin-ns cincr Hara. 

(A) / ^crividcniui^ dcr Gci^'acJisartcii diirch das Brcnucn. 

Die \ criindening der Gewilchsspecies liisst sicli aiis Untersucli- 
iingcn in Kiyosumi erkcnnen. Diese Hara wiu-de, wie schon bemcrkt, auf 



rchcr dio Eiinvirkiiiis: rtcs //ffj-rt-Brfiiuciis. 



287 



vcrschiedcncn Tlicilcii in vcrschicdcncn Zwischcnriiumcn j^ebrannt. ijic 
folgcndcn Tabcllcn bclcuchtcn in ihrcn ICrgcbnisscn dicsc l'Vaj:^c ''die 
einzeinc rrobcfl;ichc = 20 D. m. gross.) 

Kajisaka (Xordscitc^. 

{alljcilirlicJi gcbrannt) 
(Neigung = 39^) 
(Probeflachc = 20 L. m.) 

(Datum 10/4 u. 26/4) 



lateinisclicr Xamc. 


ja; aiiiseber Xamc. 


. u 


.Miscamlius sinensis Anders. 


.Susuki 


2S 




Sanguisorba Dflicinnlis 1.. 


Warcmoko 


s 




Carex Duvaliana J-r. ct Sav. 


Kesugc 


s 




liracliypcxlium silvaticum K. ot S. 


^'amakan1aiigusa 


/ 




I'teris aquilina L. 


Warabi 


5 




(ierbera anandria Scli. Uij'. 


."^enlionyari 


5 




'riialictruni minus I., var. fltauni Lccoy. 


-Vkikaramalsu 


5 




l'licp;opteris 'J'otla Mett. 


Mizoshida 


5 




-Vstilhc 'riumhert^ii .Mii]. 


Toriashishuma 


4 




Artemisia japonica Tininl;. 


Otokoyoniogi 


4 




Patrinia villosa Juss. 


Otokoeshi 


3 




Polygoiium cus]iitlatum S. et Z. 


Itadori 


3 




Carex lanccolata Itoott. 


Ilikagcsuge 


3 




Atractylatles lyrata S. el /. 


Okcra 


2 




Ccnlaiirea atrip'icitnlia (I>C.) 


Vamabnkuchi 


•» 




Senecio kramcri I'r. e( Sa\-. 


VabuiTgasa 


I 




Aster scalier 'I'liunl>. 


Sbirayamai;iku 


I 




I!upliorl)ia sielioldiana Mor. et Dee. 


NatsutCHkai 


I 




Aster Iriiiervius Ro\l>. var. japonica Max. 


Koiikiku 


' 




< Vnumda rcgab's L. var. japonica Milde. 


Zemmai 






Ani^clica decursiva Mi(|. 


Xotlako 1 






Serratula eoronata 1.. 


Tamalxiki 






Cirsiuin spicatiim Maxim. 


Yamaaiami 

1 







288 



0. Sliishido: 



Sanicula data Miq. 

Aconitum sinense S. et Z. 

Sedum l<amtscliacl"iicum Pl?cb. 

Disporium pullum Salisb. 

Cypripcdilum japonicuni Tliunli. 

Saussurea ussuriensis Max. 

Coelopleurum Gmeliiii Ledeb. 

Adenophora verticillata Fi?cli. var. vcrticillala (Fr. cl Sav. 

Saussurea affinis Spr. 

Petasites japonicus Mu[. 

Ilosta Sieboldiana Engl. 

Solidago Virga-aurea L. 

Patrinia scaliiosaefolia Linb. 

Lconvirus macrantlius Maxim. 

Smilax cliina L. 



Umanomitsuba 
Torikabuto 

Kirinsu 

Tucliikuran 

Kuniagaiso 

Kikua/ami 

Shishiudo 

Tsun'ganc ninjin 

Kilsuneazanii 

Fuki 

Tcj giliushi 

Awadachisu 

Hminaeslii 

Kisewata 

[Sankirai] 



Kajisaka (Siidseite). 

{alljaJirlicJi gcbrannt) 
(Neigung = 32-) 
(Probeflache — 20 D. m.) 
(Datum 10/4 u. 26/4) 



kiteinischer Name. 


japanischer Name. 


0^ 


Miscantlius sinensis Anders. 




Susuki 


34 


Uracliyiiodiuni sclvalicuni R. ct S. 




\'amakamojigusa 


6 


Arundinclki aiiomala Slcud. 




Ttxlasbiba 


5 


Tlialictruni minus L. var. chituni Locoy 




Akikaramatsu 


5 


Sanguisorlja officinalis I>. 




Waremoku 


4 


Potcntilla fragarioidcs L. var. ternata M 


ixini. 


Mitsubatsuchiguri 


4 


I'lectlanllius glaucocalyx Max. var. jaimnicus Max. 


1 likiokoslii 


3 


Plicgopicris Totta McU. 




Alizosliida 


3 


Artemisia japonica Tliuid). 




Oloko yomogi 


3 


I'oa trivialis L. 




llinic ieliigolsunagi 


3 



L'cbfi- (lie Kiinviikiin^' des Jlaru-linuiuu^. 



289 



Aster scabcr Tiiunh. 

Sciiccio kraincri Fr. ct Sav. 

Kupliorhia Sielioldiaiia ^^)r. ct iJcc. 

Viola J'atrinii IJC. var. chinciisis Ging. 

Taraxacum ollidnalc Wig^-. var. plauscscc.is Kocli. 

I'lcris acjuilina 7,. 

I.y-s'machia tlcthro.'clcs Duhy. 

Aster japoiiicus .Miij. 

JJisjwrum scsilc Don. 

Cirsiuni spicatum (Maxim.) 

^aussu^ca ussurieiisis Maxim. 

Polygonum cuspidatum S. ct /. 

C'o-^Ioplcurum (JmcJini Ledcb, 

Sescli Lihaiiostis Koch, var. d.iucifolia DC. 

Ixeris Tliunbcrgii A. Gr. 

ricris hicracicides I,, var. japoiiica i-l.^l. 

Viola japonica Longsd. 

Dioscorca Tokoro Makino. 

Mclandryum firmum Ruhr].. 

Senccio campcstris DC. 

Calystegia sepium R. lij-. 

Allium japonicum Rgl. 

Serratula corroiiata I,. 

Agrostis percniians Tuck. 

Lilium Maximowiczii Rgl. 

Rumcx accto?a I.. 

Lithospcrmum Zollingcri .\. DC. 

Cryptogrammc jajionica I'rautl. 

roiygonatum giganteum Diclr. var. Thunhergii .Max. 

Inula salicina I.. 

Cimicilusa foc-tidi L. var. simplex llutli. 

^'icia unijuga Al. l!r. 

Kulnis parvifolius I . 

Graminea S- 



Shirayamagiku 
Vahuregasa 
Xatsutodai 
Sum ire 
'ran|)op<> 
Warabi 
I C;)katoranoo 
Vanixshirogiku 
Ilochakuso 
Vama azami 
Kikujazami 
Itadori 
Shishiudo 
Iljukibofu 
Nigana 
Kozorina 
Kosumirc 
Oiudokoro 
Fushiguro 
Sawaoguruma 
Hirugao 
Vamarakkyo 
Tamura^o 
Xukabo 
Koiiiyuri 
Suiba 

1 lotarukazura 
TacJiishinobu 
Xarukoyuri 
Kascnsii 
Sarashinashuiua 
[Xantenhagi] 
[Xaw-ashiro ichigo] 
Graminea Sr. 



3 

2 
2 
2 
2 
2 
2 



290 



0. Shishido: 



Omiyama (Nordseite\ 

{scit J JaJirc niclit gcbrannt) 
(Neigung = 34"-) 
(Probefluche = 2 Q m.) 
(Datum 1 2/4 u. 26/4) 



latcliii-chcr Xanic. 


japanischer Name. 


(1 ' 


Miscantlius sineii'^is AiiJcrs. 


Susuki 


30 


Aster japoiiicu> Miq. 


Yamashirogiku 


5 


Carcx breviculmis R. I!r. 


Aosuge 


.S 


Carcx Duvariana Fr. et Sav. 


Kcsuge 


5 


Arundiiiella anoniala Sleud. 


Todashiba 


5 


Disporuui scsilc Don. 


Huchakusu 


4 


Serratula coroiiata L. 


Tama boki 


4 


Poteiitilla fragarioiiles 1-. var. TL-rnata Max. 


Milsubatsuchiguri 


3 


Artcncisia vulgaris L. var. indici Max. 


Yomogi 


3 


Kraunhia lloribunda (willd) Taub. 


Fuji 


3 


I'icris liicracioklcs L. vai". ja[)0!i!ca|Rgl. 


KGzorina 


3 


Adenopliora verticiUata Msch. var.ivcrtifillata (Fr. et Sav.) 


Tsuriganeniajia 


2 


Gentiaua scaljra Bge. var. ]>uL-rgcri Max. 


Rindu 


2 


Artemisia japonica Thunb. 


Otokoyomogi 


2 ' 


Agrini Miia viscidula l!g. var. japonic.i -Muj. 


Kinniizuliiki 


2 


luipliorbia Siel)oI(liana Mor. et Djc. 


Xatsutudai 


2 


Pteris atjuilina I.. 


Warabi 


2 


Senecio Kraineri l'"r. et Sav. 


\'aburegasa 


2 


Chrysanthemum sinen^e Sal), var. japonicum Max. 


Ryunugiku 




Aster scaber Thunb. 


Shirayaniagiku 




Angeh'ca decursiva -Mi(i. 


Nodake 




Clematis recta L. var. pauiculata (I'hunb.) 


Senninso 




IJrachypodium silvaticum R. et .S. 


^'amakamojigusa 




I'atrinia villosa Juss. 


( )lokoeshi 




Runic X aceto,5a L. 


Suiba 


\ 



Einleltang. 



261 



l{rachy])Otliuni japonicinn .Mir|. 


Kamojigusa 




Solidapo virga-aurea T.. 


Awadachiso 




Cirsiuni sjiicatuin Maxim. 


Yamaazami 




Calanthc tliscolor Liiidl. 


Ebine 




Saussurca iissuricnsis Maxim. 


Kikuazami 




I'etasitcs japonicus Mi(|. 


Fuki 




Cocloiilciirum fjmeliiii Ixrdcb. 


Shishiudo 




I'hytolocea acinosa Ro\b. var. esculcnta Max. 


Yamagobo 




I'cnthorum scdoidcs T-. var. chincnsc Max. 


Sawa shion 




Viola silvestris Kit. var. <^rypoceras A. C.r. 


Ta chitsu Ijosu ni i vc 




Aslilbc Thunbcryii Mif]. 


Toriasliishrmia 


! 10 


Osmunda rcgalis T.. var. jaimnica Mildf. 


Zcminai 




Dioscorca tokoro Makino. 


Onidokoro 




Lysimacliia clcthroidcs I)ul)v. 


Okatoranoo 




I'olygonum cusijidatum S. cl Z. 


Itadori 




Vil)urnum dilatalum Thunl). 


[Gamazum:] 




Oucreus glandulifcra 111. 


[Konara] 




(^)ucrcus scrrata Thuiil). 


[Kunugi] 




Rhododendron iiidicuin S\\ , v. v. Kacmi^icri Max. 


[Yamat.'^utsuji] 




Rul)iis parvifolius I.. 


[Na wash i roichigo] 




Smilax cliina L. 


[Saiikirai] 




Ixrsi^cdcza hicolor Tiirc/. 


[llagi] 




Spir.ica japonica T.. t". 


[Shimo.viiki] 





Omiyama (SiidsciteX 

i^scit 3 Jahrcn nicht f^cbrannt) 
(NciVung = 34C) 
(rrobcflachc = 2 U. m.) 
(Datum 10/4 u. 26/4) 



l,\tiinischcr .Nairn-. 


l.»I\inisciK-r Nan-.r. 


% 


Nb'soantlujs sinensis .\ndcrv. 
IMicgopteris lotta Mctt. 


Susuki 
Mizoshida 


29 
5 



292 



Aspidium aristatus Sw. 

Carex conica Boolt. 

Serratula coronata L. 

Disporum sessile Don. 

Agrimonia pilosa Ledeb. 

Clu-3-santhemum sinense Sab. var. japonicum Max. 

Carex Morrowi Boott. 

Houttuynia cordata Thunb. 

Potentilla fragarioides L. var. temata Max. 

Picris hicracioides L. var. japonica Rgl. 

Viola silvestris Kit. var. grypoceras A. Gr. 

Artemisia japonica Thunb. 

Angelica decursiva Miq. 

ArundineUa anomata Steud. 

Sodum kamtscboticum Fisch. 

Carex duvai'iana Fr. et Sav. 

Aspidium lepidocaulon Hook. 

Eupatorium Kirilowii Tuixz. 

Bracbypodium silvaticum R. et S. 

Aster scaber Thunlj. 

Angelica polymorpha Maxim. 

Euphorbia sieboldiana Mor. et Dec. 

Lactuca Thunbergiana (A. Gr.) Maxim. 

Gentiana scabra Bge. var. Buerger; Max. 

Ptcris aquilina L. 

Senecio cramcri Fr. et Sav. 

Osmunda regalis L. var. japonica Mild. 

Athyrium filix femina Rotli. 

Artemisia vulgaris I^. var. indica Maxim. 

Cryptogramme japonica Prantl. 

Patrinia scabiosacfolia Link. 

Patrinia viliosa Juss. 
Ophiopogon japonicus Kcr. 
Dianthus superbus L. 



0. Shishido: 








Hosoba kamavvarabi 


3 




Himekansuge 


3 




Tamabuki 


3 




riuchakuso 


3 




Kinmizuhiki 


3 


um Max. 


Ryunogiku 


2 




Kansuge 


2 




Dokudami 


2 




Mitsubatsuchiguri 


2 




Kozorina 


2 


. 


Tachitsul)Osumire 


2 




Otokoyomogi 


2 




Nodake 


2 




Todashiba 


2 




Kirins5 


2 




Kesuge 


2 




Orizurushida 






Sawabiyodori 






Yamakamojigusa 






Shirayamagiku 






Shiranesenkiu 






Natsutodai 






Nigana 






Rindn 






Warabi 






Yaburegasa 






Zemmai 






Meshida 






Yomogi 


\ 




Tachishinobu 






Ominaeshi 






Otokoeshi 






Yomohige 






Kawaranadeschiko 





Einleitiin^. 



293 



Saussurea ussuriensis Maxim. 

Erigeron annuus Pers. 

Lonicera japonica Thunb. 

Cypripedium japonicum Thunlj. 

Plantago major L. var. asiatica Dene. 

Crematis recta L. var. peniculafa Tliuiib. 

Iris japonica Tliunb. 

Petasites japonicus Miq. 

I.ysimachia clethroides Duby. 

Arisaema japonicum Bl. 

Vitis Thunbergii S. et Z. 

Coelopeurum gmclini Ledeb, 

Clematis japonica Thunb. 

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. 

Acanlhopanax ricinifohum S. et Z. 

Ouercu=; glandulifera Bl. 

Q^ucrcus serrata Thunb. 



Kikuazami 

Himejoon 

Nindo 

Kumagaiso 

Obako 

Senninso 

Shoga 

Fuki 

Okatoranoo 

Tennansho 

Ebizuru 

Shishiudo 

Hanshozuru 

[Xawashiroicliigo] 

[Akebi] 

[Terihanoibara] 

[Mitsubaakebi] 

[Himehagi] 

[Himeutsugi] 

[Iwayanagi] 

[Sankirai] 

[Bodara] 

[Konara] 

[Kunugi] 



Omiyama (Siidseite). 

(seit 6 Jahre nicht gebrannt) 
(Neigung = 36°) 
(Probcflache = 2 D. m.) 
(Datum 12/4 u. 26 '4') 



i.iteinischor \amc. 



Miscanthus sinensis .\nder< 



japaiiischcr Xanic. 



, 17. 



Suzuki 



294 



0. Shishidu: 



Carex duvariana Fr. et Sar. 

Poa trivial is L. 

Cryptogramme japonica Prantl. 

Viola silvestxis Kit. var. grypoceras A. Gr. 

Carex confcrtiflora Bootl. 

Trycyrtis chirta Hook, 

Polygonatuni giganteuni Oictr. var. Tliunbergii Max. 

Phegopteris Totta Mett. 

Eupatorium japonicum Thunb. 

Thalictrum minus I-. var. datum I.ecoy. 

Gentiana Zollinger! Fawe. 

Gentiana scabra Bge. var. Buergeri Max. 

Hypericum sampsoni Hce. 

Salvia japonica Thunb. var. bipinnata Fr. et Sav. 

Brachypodium silvaticum R. et S. 

Potentilla fragarioides I., var. tcrnata Max. 

Aster japonicus Miq. 

Rubia cordifolia L. var. mungista Miq. 

Gynostemma pedata Bl. 

Pteris aquilina L. 

Ophiopogon japonicus Ker. 

Aconitum fisclieri Reicli. 

Artemisia vulgaris I., var. indica Maxim. 

Patrinia scabiosaefolia Fink. 

Patrinia villosa Juss. 

Polygonum cus]iidatum S. et '/.. 

Arisaema japonicum Bl. 

I'o.i acroleuca Stena. 

Dianthus superbus L. 

Osmcirhi/.a jajicnica S. et Z. 

Scnccio Krameri ]'"r. et Sav. 

Cirsium spicitum Maxim. 

Viola japonica I-angsd. 

Clematis recta L. var. ]iaiiiciilata 'riuiiil). 



Kesuge 

Ilime ichigotsunagi 

Tachishinobu 

Tachitsubosumire 

Shirasuge 

riototogisuso 

Narukoyuri 

Mizoshida 

Hiyodoribana 

Akikaramatsu 

Fuderindu 

Rindo 

Tsukinukiotogiri 

AkinotamurasG 

Vamakamojigusa 

Mitsubatsuchiguri 

Yamashirogiku 

Akane 

Amachazuru 

Warabi 

Janohige 

Torikabulo 

Yomogi 

Ominaeshi 

(Jtokoeslii 

Itadori 

Tennansho 

Mizo ichigotsunagi 

Kawara nadeshiko 

Yabuninjin 

Yaburegasa 

Yamaazami 

Kosumire 

Senninso 



PJinloitiin;;. 



295 



Asarum caulescens Mi(|. 

Aster scabcr Thunl). 

Plectranthus inflcxus \alil. 

Clematis apiifolia DC. 

Aspiduini scuk-atuni Docll, var. japonicuni (l'"r. et Sav. 

TaraxaDuin ofriciiiale \\'igt:;. var. <.jlauceesccns Kocli. 

T-onicera japonica Thunh. 

Petasites japonicus Miq. 

Platanthcra cliloraiitha Cu-;(. 

Inula salicina L. 

Saussurca ussurricnsis Maxim. 

Bromus japonicus Thunh. 

Astilbe Tliunbergii Miip 

Calantiie rcflexa Nfaxim. 

Hedera liclix I.. 

Rubus I5ucrg(.-ri M'u\. 

Viburnmu dilatatum Tl\unb. 

Akebia quinata Dene. 

Akebia lobata Dene. 

Euonymus alala K. Kock. var. subtritlora I'V. ct Sav. 

Quercus glandulifcra 1!!. 

Rubus pahnalus Thunb. 

Aucuba iapoiiiea Tliuiil). 

Torrcya nucitVra S. et Z. 

Quercus myrsinaefolia III. 

Spiraea japonica L. f. 

Lcspeileza l^icolor 'I'lirc/. 

Smiiax cliiiia 1 . 

V'acciiiium braeteatuni Tliunb. 

Deut/ia gracilis S. ct /. 

I.itsea glauca Sicb. 

Rhododendron indicuin S\v. v,ir. Kacin]it"eri Max. 

I.indcra selicea Ul. 

Clethra barvinervis S. et /. 

Diervilla grandillora S. et '/.. 



Kan aoi 

Shirayaniagiku 

Yama hakka 

Botanzuru 

Inode 

TanpofK) 

Nindo 

Fuki 

Ginbaiso 

Kasenso 

Kikuazami 

Suzumcnochahik i 

Torias b ishoma 

Xatsuebine 

[I'uyuzuta] 

[Fuyuichigo] 

[Ganiazumi] 

[Akebi] 

[Mitsubaakcbi] 

[Komayumi] 

[Konara] 

[Kiichigo] 

[Aoki] 

[Kaya] 

[Shirakashi] 

[Shimotsuke] 

[Hagi] 

[Saiikirai] 

[Shash;uiix>] 

[Himeutsugi] 

[ShircKlamo] 

[\an\atsutsuji] 

[Kuromoji] 

[Ryobu] 

[Hakoncutsugi] 



296 



0. Shishido : 



Musadogadai (Siidseite). 

(seit 6 Jahre nicht gebrannt) 
(Neigung = 29°) 
(Probeflache = 20 D. m.) 
(Datum 1 1/4 u. 27/4) 



lateinischer Name. 


japani.scher Name. 


0/ 


Miscanthus sinensis Anders. 


Susuki 


18 


Carex breviculmis R. Br. 


Aosuge 


3 


Carex duvariana Fr. et Sav. 


Kesuge 


3 


Viola silvestris Kit. var. grypoccras A. Gr. 


Tachitsubosu mire 


3 


Artemisia vulgaris L. var. indica Maxim. 


Yomogi 


3 


Angelica anomala Pall. 


Yoroizasa 


3 


Aster trinervius Roxb. var. adustus Maxim. 


Konkiku 


3 


Disporum sessile Don. 


Plochakuso 





Viola patrinii var. chinensis Ging. 


Sumire 


3 


Brachypodium japonicum Miq. 


Kamojigusa 


3 


Aspidium dissectum Matt. 


Iloschida 


2 


Houttuynia cordata Thunb. 


Dokudami 


2 


Thalictrum minus L. var. elatum Lecoy. 


Akikaramatsu 


2 


Patrinia villosa Juss. 


Otokoeschi 


2 


Lysimacliia clethroides Duby. 


Okutoranoo 


2 


Aster japonicus Miq. 


Yamashirogiku 


2 


Viola japonica Langsd. 


Kosumire 


2 


Aster scabcr Thunb. 


Schirayamagiku 


2 


Pteris aquilina L. 


Warabi 


2 


Arimonia viscidula Bge. var. japonica"_Miq. 


Kinmizuhiki 


2 


( 'irsium spicatum Maxim. 


Yamaazami 


2 


Rubia cordifolia I., var. mungista Miq. 


Akane 


2 


I'lcctrantlius glaucocalyx Max. var. japonicus Max. 


Ilikiokoschi 


I 


Galium asprcllum Michx. 


(Miayacmugura 


I 



Senccio Kramcri Fr. ct Sav. 



Yaburcgasa 



Einleitung. 



297 



Scrophularia patriniana Wycll. 

Euphorbia sieboldiana Mor. et Die. 

Vitis Thunbergii S. et Z. 

Serratula corona ta L. 

Plantago major L. var. asiatica Dene. 

Belamacanda cbinensis Lem. 

Potentilla fragarioidcs L. var. tcrnata Maxim. 

Galanium nepalense Sweet. 

Dianthus superbus L. 

Lactuca Thunbcrgiana (A. Gr.) Maxim. 

Angelica decurtiva Miq. 

Petasites japonicus Miq. 

Clematis recta L. var. panieulata Thunb. 

Osmunda regalis L. var. japonica Mildc. 

Platanthera mandarinorum Reich, f. 

Cirsium joponicum DC. 

Akebia quinata Dene. 

Clematis apiifolia DC. 

Rosa multiflora Thunjj. 

Poly gala japonica Ilautt. 

Rubus parvifoHus L. 

Akebia lobata Dene. 

I-espedeza bicolor Turcz. 

Dcutzia gracilis S. et Z. 

Quercus glandulifcra BI. 

Smilax china L. 

Rubus paimatus 'riiUiib. 

Spiraea japonica L. f. 

Kraunhia floribunda (Willd) Taub. 

Viburnum dilatatum Thunb. 

Dicrvilla grandiflora S. et Z. 

Spiraea Thunbergii Siob. 



Hinano usutsubo 

Natsutodai 

Ebizuru 

Tamuraso 

Obako 

Iliogi 

Mitsubatsuchiguri 

Furosd 

Kawara nadeshiko 

Nigana 

Nodake 

Fuki 

Sennins5 

Zemmai 

Yamasagiso 

Noazami 

[Akebi] 

Botanzuru 

[Nuibara] 

[Hiraebagi] 

[ Xawashiroichigo] 

[Mitsubaakebi] 

[Hagi] 

[Himeutiiugi] 

[Konara] 

[Sankirai] 

[Kiichigo] 

[Shimotsukc] 

[Fuji] 

[Ganvizumi] 

[Ilakoneutsugi") 

[Tw.T\-nn.Tgi] 



> 25 



298 



(>. Shisliido: 



SuzAiriischi (Siiclseite) 

(uber 8 Jahrc nicht gebrannt) 
(Neigung = 33°) 
(Probeflache = 20 D. m.) 
(Datum 12/4 u. 27/4) 



latciiiischcr Xauie. 


japonischer jVame. 




Miscanthus sinensis Anders. 


Susuki 


10 


Carcx lanceolata Boott. 


Hikagesuge 


4 


Phegopteris Totta Mett. 


Mizoshida 




Viola silvesti-is Kit. var. gi-ypoccras A. Gr. 


Tachitsubosumire 


3 


Dianthus superbus I.. 


K aw ar anadeshiko 


3 


Txcris Thunbergii A. Gr. 


Xigana 


3 


Aspidium aristatum S\v. 


Hosoija kana\varal)i 


3 


.\rtemisia japonica Thunli. 


Otokoyomogi 


3 


Tricytis hirta Hook. 


I Iototogisus-3 


3 


.Muhlenbergia Iluegcrii Trin. 


Onezumigaya 


3 


J. ilium Maximowicizii Kegel. 


Koniyuri 


3 


Sedum Kamtschaticum Fiscli. 


Kirinso 


3 


Viola Fatrinii DC", var. ehincnsi> CWiig. 


Sumire 


2 


Carpesium ccrnuum T.. 


Sajigankuljisu 


^ 


Brachypodiuni silvaticum R. et S. 


\'amakamojigusa 


2 


Woodwardia radicans Sm. var. oriental is^l.iirs. 


Komochishida 


2 


Osmunda regalis L. var. japonica Mild. 


Zemmai 


2 


Potentilla fragarioidcs L. 


Kijimushiro 


2 


I'teris cretica L. 


Obainomotosu 


2 


Rul)ia cordifnlla L. var. nuHigisla Mii|. 


Akane 


2 


I-"uphorl)ia Sielioldiana Morr. et Dene. 


Natsutudai 


2 


("irsium spieatuni (Maxim) 


N'aniaazanii 


2 


Pteris aquilina L. 


\Varal)i 


3 


Aster japonieus MI(i. 


^'alna^l)il■ogiku 


2 


Gentiana sealira Hge. var. IJucrgeri Maxim. 


Rin.ir, 


2 



Ueber dif* Kiinvirkiiiig dcs /£ar«-Breinieii« 



299 



Chrysanthemum sinensc Sab. var. japonicum Max. 

Arimonia cordifolia L. var. mungista Miq. 

Ai-temisia vulgaris L. var. indica Maxim. 

Solidago Virga-aurca L. 

Senecio krameri ¥r. ct Sav. 

Saussurea japonica DC. 

Seseli lyibanostis Koch, var. daucitolia L'C. 

Serratula coronata L. 

Houttuynia cordata Thunb. 

Clematis japonica Tlmiib. 

Astilbc Thunbergii Miq. 

Polygonatum giganteum Dictr. var. 'Jlumbergii Max. 

Petacites japoiiicus Miq. 

Thah'ctrum minus L. var. elatum Lecoy. ' 

Cymbidium vircns Liiidl. 

Lonicera japonica Thunl). 

Inula salicina 1.. 

Atractylis lancea Thunl). 

Saussurea ussuriensis Maxim. 

Rhodea japonica Rho". 

Ligularia Ki-cmpferi S. ct Z. 

Rubus parvifolius L. 

Poiygara japonica Hautl. 

Rubus incisus 'lluinb. 

llcdcra Iich'x L. var. colcliico C. Koclt. 

Akebia lobata Dene. 

Rubus pahiiatus Thunb. 

(^^uercus glandulifcra PI. 

Deutzia gracilis S. ot Z. 

Dcut/.ia scabra Thunb. 

Dicrvilla]grandiflora S. ct Z. 

Perbcris Tlumbcrgii DC. 

I.imlcra sclice.i PI. 

Rhodvxlo'.idron in iicum S\v. v.\r. K.vnipfcri Max. 



Ryunogiku 

Kinmizuhiki 

Yomogi 

Akinokirinsj 

Yaburegasa 

Himehigotai 

Ibukibofu 

Tamuraso 

Dokudami 

Hanshozuru 

Toriashishoma 

Narukoyuri 

Fuki 

Akikaramatsu 

Shunran 

Suikazura 

Kasenso 

Okera 

Kikuazami 

Omoto 

Tsuwabuki 

[Xawashiroichigo] 

[Himehagi] 

[Nigaichigo] 

[Fuyuichigo] 

[Mitsuba-akebi] 

[Momijiichigo] 

[Konara] 

[Himcutsugi] 

[Utsugi] 

[H.ikoneutsugi] 

[Mogi] 

[Kuromoji] 

[Vanutsutsuji] 



300 



0. Shishido: 



SmiLix: china L. 


[Sankirai] 




Eurya japonica Thunb. 


[Hisakal<i] 




Lespedeza bicolor Turcz. 


[Hagi] 




Clethra barvinervis S. et Z. 


[Ryobu] 




Lit sea glauca Sieb. 


[Shirodanio] 




Spiraea japonica L. f. 


[Shimotsuke] 




Aucuba japonica Tliunlj. 


[Aoki] 





Suzuriishi (Nordseite). 

(bis 24 Meiji (1891) alljahrlich und spiiter 

im Jahre 33 Mciji (1900) gebrannt) 
(Neigung = 32-) 
(Probeflache = 20 '1a. m.) 
(Datum 13/4 u. 27/4) 



lateinischer Name. 



japanischer Name, 



Miscanthus sinensis Anders. 

Phegopteris totta Mett, 

Chrysanthemum sinense Sab. var. japonicum MaN 

Gentiana scabra Bge. var. Buergeri Maxim. 

Carex duvariana PV. et Sav. 

Carcx confertiflora Boott. 

Brachypodium silvaticum R. et S. 

Carcx brunnea Thunl^. 

Cirsium spicatum Maxim. 

Kraunhia floribunda (Willd) Taul). 

Uisporum sessile Don. 

Trityrtis hirta Hook. 

Carpcsium cernuum L. 

Clematis hcraclcifoha DC. var. stans S. et Z. 

Lactuca denlicukita Maxim. 

Batrinia villosa Juss. 



Susuki 

Mizoshida 

Ryunogiku 

Rindo 

Kesuge 

Shirasuge 

Yamakamojigusa 

Nakirisuge 

Yamaazami 

Fuji 

llochakuso 

Ilototogisuso 

Sajigankuliiso 

Kusabotan 

Yakushiso 

Otokoeshi 



12 
5 
4 
4 



Ueber die Einnirknng des 7/rtra-Biennens. 



301 



'I'linlictrum minus L. var. elaluni I.ccoy. 

Camj^iniila jiuiictata Lam. 

Snlidago \'irtja-aurca L. 

Lysimacliia clclliroides Duliy. 

I'atrinia scaljiosacfolia Link. 

Dianthus supcrlius L. 

Osmunda rcpjalis L. var. japonica MilfK-. 

ricris hieracioides L. var. japonica Re;!. 

Aristolochia K?empferi Willd. 

Serratula coronata I.. 

Iletcropappns hispidus Less. 

Cryptogramme japonica Prantl. 

Eupatorium japonicuni Thunl). 

Inula salicina L. 

Woodwardia radicans Sm. var. nricntalis Lurs. 

Asterom?ea indica BI. 

Clematis recta L. var. paniculata Tliunl). 

Petasites japonicus Miq. 

Aster scalier Tluml). 

Senccio Kramer! Fr. el Sav. 

Potentilla fragarioidcs L. var. ternata Maxim. 

llouttuynia cordata Thunl). 

Cocloplcurum gmelini Ledeb. 

Astillic Thunbergii Micj. 

I'Aipliorhia sieboldiana Mor.'et Hie. 

Artemisia japonica Thunh. 

Soseli i.ihannstis Kocli. var. daiicilolia DC. 

Plantago maj<M- L. var. asiatica IVne. 

Ligularia Ka-mplViM S. el '/.. 

C'ymhidivim \ii-etis I.indl. 

jMlium japonicum Kgl. 

Lonicera japonica Tlumh. 
Rubus Buerger! Miq. 
.*^]iira\T japonic,! L. f. 



Okikaramatsu 

Hotarubukuro 

Akinakirinso 

Okatoranoo 

Ominaeshi 

Ka waranadesh iko 

Zemmai 

Kozorlna 

Oba umanosuzukusa 

TamurasG 

Yamajlnogiku 

Tachishinobu 

Hiyodoribana 

Kasenso 

Komochishida 

Yomcna 

SenninsG 

Fuki 

Shirayaniagiku 

Yabiircgasa 

Mitsubat<uchiguri 

Dokudami 

Shishiudo 

Toriasbishoma 

Natsutiklai 

Otokoyomogi 

Ibukiliiifu 

Obako 

Tsuwabuki 

Sliunran 

Vamirakkvo 

Nindo 

[Fuyuichigo] 

[Xawashiro"ch'g»"»} 



302 



0. Sliishido: 



Rubus parvifolius I.. 


[Shimotsukc] 


Polygaia japonica Hautt. 


Himehagi 


Akebia lobata Dene. 


Mitsuba-akcbi 


Ficus foveolata Wall. 


Itabikazura 


Quercus glandulifera I>1. 


Kouara 


Quercus serrata TI;um1). 


Kunugi 


Smilax china L. 


Sankirai 


Rubus palmatus Tliunh. 


Kiichigo 


Clctlira barvinernis S. ct Z. 


Ryobu 


Diervilla gi-andiflora S. ct Z. 


ITakoncutsugi 


Dculzia gracilis S. et Z. 


Mimcutsugi 


IVulzia scabra Thunb. 


Utsugi 


Lindcra sclicera Bl. 


Kuromoji 


Torreya nucifcra S. ct Z. 


Kay a 


Aucuba japonica Thunb. 


Aoki 


Eurya japonica Thunb 


Hisakaki 


Spiraea Thunbergii Sicb. 


Iwayanagi 


Ardisia crenata Sims. 


Manryo 


Rhododendron indicuin S\v. var. K.xmpfcri Max. 


Yamatsutsuji 


Maclcya cordata R. T'r. 


Chanpagiku 


Poa trivialis L. 


Himeichigotsui 


Pteris serrulata I., f. 


Inomotosu 


Pteris aquilina L. 


Warabi 



2K, 



Sannodai (Nordscitc). 

(bis 24 Meiji (1891) alljahrlich und im 

Jahrc 33 Meiji (1900) gebrannt) 
(Neigung^35^) 
(Probefliiche = 20 G. m.) 
Datum 14/4 u. 27/4) 



iatcun'schcr Name. 


japanischcr Name. 


0/ 
/o 


Miscanthu; sinensis Anders. 


Su'^uki 


16 



Ut'lx'i- (lie p]iJMvirkuiii|ir <l«'s J/arfi-lircuucus. 

Aosugc 



303 



Caicx Ijrcviculmis R. I!r. 

Picris hicracioidcs L. var. japoiiica Rfjl. 

Carcx lirunnca Tluinl). 

Viola silveslris Kit. var. tjrypoccra?; A. Gr. 

Tricyrtis liirta Ilfiok. 

(jcntiana scabra B;4C. var. Bucrgcri Maxim. 

Salvia japonica 'i'liunl). var. 1»ipinnata Fr. ct Sav 

Carpcsium ccnuium I.. 

Artemisia vulp;aris 1 . var. indisa Maxim. 

Aster japonicus Miq. 

Craw furdia iitcrygocalyx Hcmsl. 

Caiex conica lloott. 

Polygonaliim lasiantlium Maxim. 

Lactuca danticulata AFaxim. 

Aspidium laccrum S\\ . 

Asarum Blumci Ducli. 

Eupatorium japoiiicum Thunl). 

Aspidium crytlirosarum Eat. 

Aristolochia Kacmpfcri Wiild. 

Osmunda rcgalis L. var. jajonica Mild. 

l\itrinia villosa Juss. 

Astilha Thunbergii Micj. 

Cirsium spicatum Max. 

Senecio Kramcri Fr. ct Sav. 

Cypripcdilum japonicum Tluml). 

C'innbidiuin vircu^ Lindl. 

Patrinia scahiosaefolia Link. 

Clematis japonica Tluinli. 

Lilium auratum Lindl. 

Houttuynia cordata 'Iliunh. 
rtcris aquilina L. 
Brachypodium silvatieum R. ei S. 

Liguralia Kivmpfori S. ct Z. 

Rluxlea japonica Rhot. 



Kozoriiia 


4 


Nakirisugc 


4 


TachiLsubo5uniirc 


3 


HototogisuFo 


3 


Rindo 


• 3 


Akinotamuraso 


3 


Sajigankubisij 


3 


Yomogi 


3 


Yaraashirogiku 


2 


Tsururindo 


2 


Himekansuge 


2 


Miyamanarukoyuri 


2 


Yakushiso 


2 


Kumawarabi 


2 


Kan-aoi 


2 


Hiyodoribana 


2 


Benishida 


2 


Oba-umanosuzukusa 


2 


Zcmmai 




Otokoeshi 




Toriashishoma 




Yamaazami 




Yaburegasa 




Kuma^aiso 




Shunran 




Omi.uicMii 




Hanshozurn 




I'amayuri 




Pokudami 




Warabi 


\ 


Vamakaniojigtisa 


Tsuwabuki 




Onioto 





504 



0. Shishido: 



Clematis recla L. var, paniculata (Thunl)). 

Mitchella undulata S. ct Z. 

Lindera umbellata Thunl). 

Quercus glandulifera BI. 

Rhus succedanea L. 

Rosa Wichuraiana Crep. 

Hedera belix L. var. colchica C. Kcch. 

Akebia quinata Dene. 

Spa-£ea japonica I., t. 

Trachelospermum jasminoides Lemairc 

Ficus fovcolata Wall. 

Ruhus Buerger! Miq. 

Akebia lobata Dene. 

Vaccinium bractcalum Thunb. 

Pieris japonica D. Don. 

Illicium anisotum L. 

Thea japonica (L.) Wois. 

Eurya japonica Thunb. 

Rubus palmatus Thunb. 

Rosa multiflora Thunb. 

Rhododendron indicum S\v. var. K«?mpfcri MaN 

Osmanthus aquifolium B. ct H. 

Lindela selicea Bl. 

Diervilla grandiflora S. ct Z. 

Deutzea gi-acilis S. ct Z. 

l\ubus parvifolius L. 

Smilax china I>. 

Litsca glauca Sicb. 

Zanthoxylum schinnifoliuni S. ct Z. 



Senninso 

Tsuruariclushi 

[Kanakuginoki] 

[Konara] 

[Tsutaurushi] 

[Terihanoibara] 

[Fuyuzuta] 

[Akebi] 

[Shimotsuke] 

[Teikakazura] 

[Itabikazura] 

[Fuyuichigo] 

[Mitsubaakebi] 

[Shaslianpo] 

[Asebi] 

[Shikimi] 

Tsubaki 

Hisakaki 

Kiichigo 

Noibara 

Yamatsutsuji 

Hiiragi 

Kuronioji 

Hakoneutsiigi 

Himeutsugi 

Shimotsuke 

Sankirai 

Sh'rodamo 

Inusansho 



Leber ic Eiiiwirkimj? des Jlara-Bronntns. 



305 



S;innodai (Sudscite). 

(bis 24 Meiji (1891) alljiilirlich und im 
Jahre 33 ?*Ieiji (1900) gcbrannt) 
(Neigung^3i^) 
(Probeflachc = 20 G. m.) 
(Datum 14/4 u. 27/4) 



lateinischcr Xainc. 



japanij^cht-r Name. 



Miscanthus sinensis Anders. 

O.xalis corniculata L. 

Carex brevicurmis R. Br. 

Viola Keiskei Miq. 

Carex confertiflora Bott. 

I'hagopteris totta Mett. 

Disporuni scsscle Don. 

Carex brunnea Tluinb. 

Salvia japonica Thunb. var. Buerger! .Max. 

lirachypodiuni silvatieum R. et S. 

Viola silvcstris Kit. var. grypoceras A. Cr. 

Carpesium abrotanoides L. 

Kupatorium japonicur.i Thunb. 

Crepis japonica Bentli. 

Bothriosperniuni tenelluni Fisch et Mey. var. aspergoiile> Max 

.Vster japonicus Mi(i. 

Kupatorium Kiril^w ii Turcz. 

Cirsiuni spicatuni Max. 

Euphorbia sicboldiana Mor. et Dec. 

.\spidiuni aristatum Sw. 

I'eracarpa circa'oides II. Kee. 

Artemisia vulgaris L. var. intUia Maxim. 

^'enecio KranK-ri IV. et Sav. 

Lactuca debilis ('I'liunb). Maxim. 



Susuki 

Katabami 

Aosuge 

Marubasumire 

Shirasuge 

Mizoshida 

HGchakuso 

Xakirisuge 

Akinotamuraso 

Yamakamojigusa 

Tachitsubosumire 

Vabutabako 

Hiyodoribana 

Oiiitabirako 

I lanaibana 

Vaniashirogiku 

Sawahiyodori 

Vamaazami 

Natsutodai 

Hosoba-kanawarabi 

Tanigikyo 

Yomogi 

Yabuncg-aSxi 

Jishibari 



14 
4 



\o6 



0. Shishido: 



I'teris aquilina L. 

Asplenium liniolatum Tliuiib. 

Cypripidilum japonicum Thuiilj. 

Gnaplialium multicej-s Wall. 

Rubia cordifolia L. vai-. Mungista Miq. 

Thalictrum minus L. var. elatuni Lecoy. 

Cryptogramme japonica Prantl. 

Cirsium japonicum DC, 

Petasites japonicus Miq. 

Patrinia villosa Juss. 

Astilbe Thunbergii Miq. 

Cremastra Wallichiana Lindl. 

Gentiana scabra Bge. var. Puergiri Maxim. 

Lactuca denticulata JNIaxim. 

Cymbidium virens Lindl. 

Rliodea japonica Rhot. 

Clematis apeifolia DC. 

Akebia quinata Dene. 

Rubus incisus Thunb. 

Duchesnea indica Fock. 

Clematis japonica Thunb. 

Sambucus raccmosa P. 

Myrica rubra S. et Z. 

Eurya japonica Thunb. 

Rhododendron indicum Sw. var. Kcempferl Max. 

Torreya nucifcra S. el Z. 

Pieris japonica D. Don. 

Deutzia gracilis S. et Z. 
Lindera selicea Bl. 
Qucrcus glandulifera Bl. 
Thea japonica (L.) Nois. 
-Machilus Thunbergii S. et '/. 
Rosa multiflora Thunb. 
Acanthopanax ricinifolium S. et Z, 



Warabi 

Ilerashida 

Kumagaisu 

Chichikogusa 

Akane 

Akikaramatsu 

Tachishinobu 

Noazami 

Fuki 

Otokoeshi 

ToriasIiishGma 

Saihairan 

Rindu 

Yakushiso 

Shunran 

Omoto 

Botanzuru 

[Akebi] 

[Nigaichigo] 

[Hebiichigo] 

[Uanshozuru] 

[Niwatoko] 

[Yamamomo] 

[Hisakaki] 

[YamatsutsujiJ 

[Kaya] 

[Asebi] 

Plimeutsugi 

Kuromoji 

Konara 

P'subaki 

Tabu 

Noibara 

Bodara 



1 



Ueb(r die Eiinvirkiing des //aive-Bn-iiiU'iis. 



307 



Litsea glauca Sich. 
Rul)u.s palniatus Tliunl). 
DicTvilla t^raiHli/loia S. et '/,. 
C'lc-inatis recta L. var. as'atica Dene. 
Lonitcra japjiiica Tliunl). 



Shiroflamo 

Kiichigo 

Hakoneutsugi 

Scnninso 

Nind«"> 



Nanamagari. 

(seit 10 Jahrcn nicht c,febrannt) 
(NeiiTunc? = 35-) 
(Probcflache = 20 n. m.) 
(Datum 15/4 u. 26/4) 



latcinisclicr Name. 



Miscanthus sinensis Anders. 

Ark-niisiii japonica '1 liuni). 

Palygonaluni oniciiiale All. 

Carcx inc^sa Koott. 

Carcx diivariana Fr. et Sav. 

Crypt otjraninie japonica Prantl. 

Picris hieratioidos L. var. japonica Rgl. 

Chrysanthemum sincnse Sab. var. ja|ionicum N?ax, 

Viola japonica Lan;^'sd. 

Aristolachia KaomplVri Willd. 

nispornin scsilc Don. 

Carcx conica ISoott. 

Brachypo iium silvalicum R. ct S. 

Serratuia coronala I,. 

Aster japonicus i\li.|. 

'I'lialiclrinn minus L. v.ir. datunj Lcvoy. 

Potcnlilla fraj^arioidos I,. 

I'olygonatum i^M^Miikum Didr. var. 'riiunl.cp.^ii Max. 

Sodum kamlschalicuni Kisch. 



japanischcr N.ime. 



Susuki 

Otokoyomogi 

Amadokoro 

Kawarasuge 

Kesuge 

Tachishinobu 

Kozorlna 

Ryunogiku 

Kosumiic 

( )l>a-umaiiosusukus.i 

Ilochakuso 

IlinK'kansuge 

Vamakainojigus^i 

Tamura^o 

Yain.ashiivgiku 

Akik;iranKitsu 

Kijimushia> 

Narukoyuri 

Kirinso 



308 



O. Sliishido: 



rteris aquilina L. 

Angelica dccursiva Miij. 

Cirs'um spicatum Maxim. 

Euphorbia Sicboldiana Mor. et Dec. 

Astilba Thunbergii Miq. 

Clematis recta L. var. paniculata Thunb. 

Patrinia villosa Juss. 

Osmunda regalis L. var. japonica Milde. 

Potentilla fragarioides I., var. ternata Maxim. 

Artemisia vulgaris L. var. indica Max. 

Saussurea ussui-iensis Max. 

Senecio Krameri Fr. et Sav. 

Arisacma japom'cum Bi. 

Lithospermum zollingeri A. DC. 

A'^arum Biumci Duch. 

Cimicifuga foetida L. var. simplex I lath. 

Lonicera japonica Thunl). 

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. cnpitata Miq. 

Plectranthus glaucocalyx Max. var. japonicus M sx. 

Rubus palmatus Thunb. 

Rubus parvifolius L. 

Akebia lol)ata Dene. 

Vicia unijuga Al. Br. 

Enonymus alata K. Koch. var. sublriflora Fr. el Sav. 

Spinra japonica L. f. 

Oucrcus glanduilfera Bi. 

ICurya japonica Thunl). 

Diervilla grandiflora S. et Z. 

Viburnum dilatatum Thunl). 



Warabi 

Nodake 

Yamaazami 

Natsutodai 

Toriashishuma 

Senninso 

Otokoeshi 

Zemmai 

Mitsubatsuchiguri 

Yomogi 

Kikuazami 

Yaburegasa 

Teiinansho 

Hotarukazura 

Kan-aoi 

Sarashinashoma 

Nindo 

Fuki 

Akinotamuraso 

Kawaranadeshiko 

Yamayuri 

Rindo 

Suzumenohiye 

Yamahakka 

[ Momiji baichigoj 

[ N^a washiroichigo] 

[Mitsubaakclii] 

[Nantenhagi] 

[Komayumi] 

[Shimotsuke] 

[Konara] 

[Ilisakaki] 

[1 lakoneutsugi] 

[Camazumi] 



Ueber die Kiinvirkiiii^ dos J lava -lircmu'iiH. 



309 



Miicliilus Tliuiiltcrj^ii S. cf /. 




[Tabu] 1 




Cli^'tlira liarv iiiervis S. et Z. 




[Ryobu] 




Sniilax china L. 




[Sankirai] 




Qucrcus acuta 'I'liunl). 




[Akagaslii] j 




Qucrcus niyrsiiiacfolia l!l. 




[Urajirogashi] 1 




Rosa niullillcira 'I'liunb. 




[Noibara] | 




1\Iio(1ik1cii(Iioii iinlicum Sw. var. 


Kivmpfeii Max. 


[Valnatsut^uii] 1 




l\ul)us palmalus 'I'liunli. 




[Kiicliigo] 1 




Liiidcia sclicca HI. 




[Kuromoji] 




Spiriva Tliunljorgii Sicli. 




[I\vayanat;i] 





Musaclogadai (Nordscite). 

(iiber 10 Jahrc nicht ijebrannt) 
(Ncigung = 32^'^)' 
(rrobcn;lchc = 20 LI m.) 
(Datum 1 3/4 u. 27/4) 



latcinischcr Name. 


japaiiisclicr Name. 



/ 


Miscanthus siiieiiMs Aiulcrs. 


Susuki 


s 


Carcx Rini^oKliana Hoolt. 


Juzusugc 


4 


IVilya Sc.iiulens Sell. Hip. var. ovata Maxim. 


Koyaboki 


4 


HracliyiMHlimu .silvaliciim R. ct S. 


Va niaka mojigusa 


4 


riicgoplciis ToUa Mctt. 


Mizosliiiia 


3 


Carcx lUivarl.uia Vv. ct Sav. 


Kcsugi' 


3 


\'iola silvc-^tii- Kit. var. i;rapoccras .\. lir. 


Tachitsubosumirc 


3 


Carcx hrumiea TIuimI'. 


Niikirisugc 


3 


Clirysaiillicmum siiicnsc Sab. var. ia|>onicimi M.ix. 


Ryunogiku 


3 


Aslcr trincrvius Roxh. var. aihisliis Max. 


Kongiku 


2 


Tricyrtis liirta Hook. 


1 lototogisusfi 


2 


Saussurca japoiiica 1 K'. 


HinK'higotai 


- 


X'iola Paliinii IK", v.ir. chiiu-ii-is tliM^'. 


Suiuiiv 


2 


Ixcris 'riiuiibiTi;!! .A. llr. 


Nigana 


2 



3IO 



0. Shishido: 



Arimonia cordifolia L. var. mungista Miq. 

Salvia japonica Tliunb. var. bipinnata Fr. et Sav. 

Aristolochia KaDinpfcri Willd. 

Gentiana scabra Ege. var. Luergeri Maxim. 

Angelica clccursiva Miq. 

Cirsium spicatum Maxim. 

Woodwardia radicans Sw. var. japonica I iirs. 

Polygonum cuspidatum S. et Z. 

Aster japonicus Miq. 

Senecio I\j-ameri Fr. et Sav. 

Cirsium japonicum DC. 

Totcntilla frngnrioidcs I., var. ternala_Maxini. 

Astilbe Thunbergii Miq. 

Clematis recta L. var. paniculala Thunb. 

Thalictrum minus I., var. datum Lecoy. 

Artemisia vulgaris L. var. indica Maxim. 

Lonicera japonica Thunb. 

riantago major L. var. asiatica Dene. 

Lysimachia clethroides Uuby. 

Lilium auratum Fintll. 

Liguleria Kx'mpfcri S. et Z. 

Relamacanda chinensis Lem. 

Calanthe discolor, Lindl. 

Clematis heracleifolia DC. 

Saussurea ussuriensis Maxim. 

Ilouttuynia cordata Tliunb. 

Clematis heracleifolia DC. var. Stans (S. el /.) 

Polygonalum gigantcum Diclr. var. Thunbergii Max. 

I'ctasitcs japonicus Miq. 

Taraxacum officinale Wigg. var. glauciscens. Koch. 

Iledera helix L. var. colehica C. Koeh. 

Akebia lobala Dene. 

Kubus parvifolius I.. 

Rubus lluergeri Mii], 



Kimmizuhiki 

Ahirotamuraso 

Oba-umanosuzukusa 

Rindo 

Nodake 

Yamaazami 

Komochishida 

Itadori 

Yamash'rogiku 

Yaburcgasa 

Noazami 

Mitsubatsuchiguri 

Toriashishoma 

Seniiinsu 

Akikaramatsu 

Yomogi 

Nindu 

Obako 

Utokoeshi 

Yamayuri 

Tsuwaliuki 

Iliogi 

Ebineran 

Tsuriganeso 

Kilcuazami 

Dokudami 

Kusabotaii 

Narukoyuri 

Fuki 

Tanpopo 

Fuyuzuta 

Mif'^ubuakebi 

Nawashiroich'go 

I'uyuichigo 



Uebor die Eiinvirkmig dfs I/ara-Bnuuom. 



311 



Clematis jaioiiica Tliunli. 
Acantliopniiax si>iiiosuni Miij. 
Traclielospcrmum jasminoidcs Lciiieire. 
Cclastrus articulatus Tliunli. 

Eurya japonica Thuiil). 

(j)uercus myrsinacfolia l!l. 

Oucrcus glandulifcra IJI. 

Qucrcus scrrata TIiuiil). 

Torrcya nucifcra S. et Z. 

Viburnum dilatatum Tlniiil). 

Machilus Thunbcrgif S. ct Z. 

I-iiidcia sclicca Bl. 

Dcutzia gracilis S. et Z. 

Dicrvilla grandinora S. ct Z. 

Rosa multi/loin Tliunh. 

Rubus 1 almatus Tiiunb. 

Eurya ocliiir.ccac Szysz. 

Ardisia crcnata Sims. 

Spirtca Thunbcrgii SicI). 

Rliododcndron indicum Sw. v;u-. K;viiipf(.ri Max. 

Rubus parvifolius L. 

Aukuba japonica Tliunb. 

Vaccinium brarlcatuin 'lliuiib. 

Lcspcdcza bicolor Turcz. 



I Ilanshozuru 
Ukogi 

Teikakazura 
Tsuruumcmodoki 
Ilisakaki 
Urajirogaslii 
Kojinra 
Kunugi 
Kaya 
Gamazumi 
Tabu 
Kuromoji 
Ilimcutsugi 
IlakoiicufsLgi 
Xolliara 
Kiichigo 
Sakaki 
MauryG 
Iwayanagi 
Vamafsutsuji 
Shimolsukc 
Aoki 

Sbashanpo 
Ilagi 



Aus den o-civcbcncn Tabcllcn ist crkcnntlicli. class Artcn und Zahi 
</rr Pjlan:~cn ai.f den l<l;ichcn dmch das h;uiri<,rc Ihcumu dor Uant nnch 
uiul nach xcnuiiuk-it wcrdon. Auf jcdcr Ilara kommt das Susiikl odcr 
kayao-rass ini Xokxmy- vor. wcil cs ciii starkcs Anpassmij^svcrmrycii hat 
und uhcrall iiocli ocdcilil. wo dor Hodcn durch Zufall vcrvviistct mid andcrc 
(icwiiclisc nicht inchr -ut xu wachson vc-.mr.-cn. c. macht cbcn die 
t^crinnrstcn Anspriichc an ilon l^>dcii. 

Aufsckcn ovliranntcr /A../-., f;dlt iK r Frocuitsal/. a-. Kaya ab. daj;cgcn 
luvKt man orossc Mcn-cn von nauni- und Ilalbbaiim-Artcn. namcntlich 



312 



0. Shishulo: 



Ouercus gland ulif era, sowie auch mancherlel Grassarten, wclchc man auf oft 
gebrannter Hara kaum antreffcn wird. Hara-VtoAcw wclchcr liingerc Zeit 
nicht gcbrannt wird, geht in seine urspriinigliche Vcgetationsform (den 
Wald) wieder liber. 



Sengcn (Nordseite). 

(Urwald) 

(Neigung = 35°) 

(Grassarten der ganzen Nordseite) 

(Datum 15/4 u. 28/4) 



latcinisclier Name. 


japanischer Name. 


Ordnung. 


Viola silvestris Kit. var. grypoceras A. Gr. 


Ta ch i t su Ijosu m i re 




Saxifraga conlusaefolia S. et Z. 


I )aimonjiso 




I'renanthes acerifolia Maxim. 


Fukuwoso 




Cardiamlra altevaifolia S. et Z. 


Kusaajisai 




Carex Ijruunea Tliunli. 


Nakirisuge 




Tricyrtis hirta Hook. 
Ligularia japonica Less. 
J'olygonalum lasiaiilhum Maxim. 


Ilototogisuso 

Ilankaisu 

Miyamanarukoyuri 


nach der 

Haufigkeit in 

absteigender 

Skala geord- 

nct. 


Aspidium kiccrum S\v. 


Kumawaralji 






lldoiiiopsis japonica Maxim. 


Shirohana-shiljoliakama 






Aconitum Fisheri Reich. 


Torikabulo 






( )i)hiopogon japonicus Ker. 


Janoliige 






AinsliL\;a acerifolia Sell. Bip. 


Momijihaguma 






Carex Morrowi l!oo 1. 


Kansuge 






Angerica ixjlymorplia Maxim. 


Siiirmeseiikiu 






Aspidium crytlirosorum l'"at. 


Ijenishida 






Ca: ex duvariana Fr. et Sav. 


Kesuge 


' 




Disporum sesile Don. 


I locliakuso 




llymenopliyllum Ijariialuni Ikilc. 


Kuycd<okes]unol):i 




Senccio Kramei-i Fr. et Sav. 


\'aliuregasa 




Astill)e 'I'liunljergii Mi(p 


'ioriashishCima 







Ueber die Eiinvirkiin^ des //rnYf-Brcnneiis. 



313 



Gentlana scabra Bge. var. Buergeri Maxim. 

Aster japonicus Miq. 

Anemone hepatica L. 

Cacalia dclpliiniifolia S. ct Z. 

Carex conica IJoott. 

Cirsium spicatum Maxim. 

Cypripcdilum japonicum Thuiil). 

Ilosta Cfcrulea (Andr) Tratt. 

Cimicifuga japonica Spr. 

Peris cretica I.. 

Clematis japonica Thunl). 

Calantha rcflexa Maxim. 

Cypripcdilum dcbilc Rciih. f. 

Arisaema jiponicum 111. 

Asarum ISlumci Duch. 

riagiogyria cnphlcbia Mett. 

Calanthe discolor, Lind!. 

Lycopodium serratum Thunl). 

Cymbidium virens Liiidl. 



Rindo 

Yamashirogiku 

Suhamaso 

Momijigasa 

Ilimekansuge 

Yamaazami 

Kumagaiso 

Giboihi 

Kikcnshoma 

Obainomotoso 

I lanshozuru 

Natsuebine 

Koatsumoriso 

Tennansho 

Kanaoi 

Kijinoo 

Ebinc 

Togeshiba 

Shunran 



Scngcn (Siidscite). 

(Urwald) 

(Ncigung = 34°) 

(Grassartcn. in der i^aii/.cii Siid.seite) 

(Datum 17/4 u. 28/4) 



l.ilcinischer X.iu 



japanischor Name. 



< >nlnunc. 



Ainslirca accrifolia Sch. K\^. 
Carcx conica Roott. 
Aspidium aristatum S\\ . 
Ant;clica polymor[iha Ma\im. 
SaxilVas^a ci^ntiisiefolia S. el /. 
Carpesium ccrmium L. 



Momijiliagunia 

Ilimekansuge 

llosolu kana\vaml»i 

Shirancscnkiu 

Dainionjiso 

Sajigankubiso 



nach tier 
Ilauligkcit in 

Sk.iLi gcwrxl- 



314 



0. Sliishido: 



Caidiandia alternifolia S. ct Z. 




Kusaaji.sai 






Ophiopogon japonicus Kcr. 




Janohige 






Cryptogramme japonica rranll. 




Kanshinobu 






Lilium curdifoliuni Thuiih. 




LTbayuri 






Woodwardia ra-licans S\v. var. orientals Liirs, 


Komochishida 






Asaruin Bluiiiei JJucli. 




Kanaoi 






Arisaenia japonicum I!l. 




Tennansho 






Hosta Sieboldiana (11. K.) Engl. 




Togiboslii 






Polygonatum lasianthuni ,Sat. Maxim. 


Miyamanarukoguri 




Adianthum monochlamys Sat. 




Hakoncshida 




Carex Morrovvi Boott. 




Kansuge 




Calanthe discolor Lindl. 




Ebine 




Cirsium spicatum Maxim. 




Yamaazami 




Cymbidium virens Lindl. 




Shunran 




Astilbe Thunbergii Miq. 




Toriashishoma 




Iris japonica Tlmnb. 




Shaga 




Setiecio Krameri Fr. et Sav. 




Yaburegasa 




Aster japonicus Miq. 




Yamashirogiku 




Disporum sessile Don. 




Tlochakuso 




Ciniicifuga foctida L. var. simplex Iluth. 


Sarashinashuma 




Anemone hcpalica L. 




Suhamaso 




Solidago Virga-aurea L. 




Awadachiso 




IIctero[)appus hispidus Less, var. 


isochaetus Fr. et Sav. 


Yamajinogiku 




Patriiiia scabiosacftjlia Link. 




Oniinaeslii 




Chimapliila japonica Miq. 




Umegasasij 




Rhizogonium Dozyanum Lacost. 




Itacliinoshippo 




Usnea longissima Acli. 




Saruogase 





Iiii Wakl " Scni^en " gibt cs also nur gcring-c Mcngcu von Gra.ss aus 
natiirlichcn Grundcn unci jcnc Lichtgriiscr, wclclic auf jiihrlicli i^cbrannter 
I lani vorhcri'schcn, k<')nniii liicr niclit c^cfundcn wcrdcn. 

Ik'ini Kintritt in cincn q^cschlosscncn Wald wird man wahrnchmcn, dass 
fast kcin Grass auf (k-ni Waidbodcn sick llndct, wiUtrcnd in gcringer 
gcscklosscncn Bestandcn das Grass sick mckrt. Dcr Griuid davon ist, darin 



Ueber die Eiinvirknii^ <les //rtra-BreniiPns. 



315 



zu suchen, class die Sonncnstrahlen in gcschlossenem Waldc, das ganzc Jahr 
hindurch den Boden wenig erreichen konnen. Die Griiser, welchc in der 
jungeren Zeit der Bestiindc noch zahlreich sind, wcrden durch den sich 
verdichtenden Schluss der Bestiindc von Jahr zu Jahr wenigcr, da ihnen 
die zur Existenz nothige Lichtmenge allmilhh'g zu fehlcn beginnt; Aus 
gleichen Griinden kommcn au{ der Hara, welche iiber 10 Jahre nicht gebrannt 
ist und einen grossertn Antheil an l^aumen Strauchcrn anfweist, verhaltniss- 
miissig wenigc Griiser vor. 

I. Vcrinindcning der GciviicJisartoi. 

Die Tabcllen zcigen ferner dass die Menge der Gewiichsspccies auf 
oftmals gebrannter Hara abnimmt, wiihrend sic in der selten gebrannten 
Hara zunimmt, bis auf einer Hara die Baumflora vorherrschend wird. Ge- 
wiichsarten, welchc durch hiiufiges Brenncn auf einer Hara an Zahl 

abnehmen, sind : 



Disporum sessile Don. 

Salmia japonica var. bucrgcri M;u\im. 

Pims hieracioides var. japonica Rge. 

Carex brunnca Thuiil). 

Tupatriinii kirilowii Furcz. 

I'Aipotrium japonicuiii 'riuml). 

A-^aruni blunici Duch. 

Polygonatuni giganlcimi var. tlunil)ergii Max. 

Clematis hcracleifolia var. Staiis. 

Chrysanthcnnini sinense var. japonic.i .M,i\. 

Ixcris thunbcrgii A. (ir. 

Artemisia japonica Tluiiih. 

Kraunlii.i tloribmula 'I'aub. 

Diantiius svipcrbus 1-. 

Tricyrtis liii-ta I look. 

rhagoptcris tolta Mctt. 

Aster japonicus Miq. 



IlOchakusi'j 

.rVkinota murasG 

Kozorina 

Xakirisuge 

Sawahiyodori 

H iy odor! 1x1 na 

Kanaoi 

Narukoyuri 

Kusalwtan 

Ryunogiku 

Nignna 

(^tokoyomogi 

[Fuji] 

Kawaraiwdcshiko 

I lototo^isuso 

Mizoshida 

Va nushirogiku 



3i6 



0. Sliisliido: 



C;uex conica Boott. 
Potentilla fragarioides L. 
Carpesium cernuum L. 
Cardiandra altcrnifolia S. et Z. 
Artemisia vulgaris var. indica Max. 
etc. 



Ilimekansuge 
Kijimushiro 
Gankubisu 
Kusaajisai 
YoniogI 
etc. 



Unter dcrn Brenncn niclit Icidcnde Geivdchsartcn. 



Sic sind folp-ende 



Miscanthus sinensis Anders. 


Siisiiki 


Potentilla fragarioides var. ternata Max. 


Mitsubatsuchigm-i 


Serratula atriplitifolia B. et H. 


Kumatoribokuchi 


Sanguisorba officinalis L. 


Waremoko 


Tlialictrum minus \-ax. elatum Lecoy. 


Akikaramatsu 


Lysimachia clethroides Duby. 


Okatoranoo 


Plectranthus inconspicuus Miq. 


Yamahakka 


Pteris aquilina P. 


Warabi 


Senccio krameri Fr. et Sav. 


Vaburegasa 


Polygonum cuspicUitum S. et Z. 


Itadori 


Euphorbia siebo'diana !\Ior. et nic. 


Natsutodai 


Impcrala arundinaceae var. Koenigii. 


Chigaya 


Astilbe thunhergii Miq. 


'roriashislioma 


Gerbera anandria Seh. Pip. 


Sebonyari 


Athyiium nipponicum S. et Z. 


Inuwarabi 


Atractylodes lyrata S. et /. 


(3kera 


Dioscorea tokoro Makino. 


Onidokoro 


Arundinella anoniala Stcud. 


Todashiba 


etc. 


etc. 



diesc Grassartcn .sind cbcn jcnc Sichtgriiscr, wclchc nur auf alljiihrlich 
gcbranntcr Ilara sowic audi auf andcrcm bcschadigtcn z. B. verhagelten 
Eodcn noch gcdcihen. 



Ueber die Eiinvirkanir dcs //^nYt-Brciiiiciis. 



317 



(B) Grasartoi dcj- vcrscJiicdcncn Standortc, 



I . Grasartoi aiif dcin Riickcn dcr Bcrj^c. 



Gcrbera anandria Sch. I!i]). 


SeiiliOnyari 


Sanguisorba officinalis L. 


\VarLTnf)ko 


Serratula atrii)Iicifolia ];. ct 11. 


Kumatoribokuchi 


Potentilla fragarioide^ var. tcrnata Max. 


Mitsubat'^uchiguri 


Pteris aquilina L. 


Waralii 


Lysimachia cletliroidcs Duby. 


< )katoranoo 


Imperata arundinaccce var. Kocniijjii. 


Chigaya 


Astilbc thunbcrgii Miq. 


'roria.<;hishunia 


Brachypodium silvaticuni K. et S. 


Vamakamojigu>« 


Euphorbia sicboldiana Mor. ct Die. 


Xatsutodai 


Euphorbia ( Inoei Fr. ct Sav. 


Takatodai 


.\ruii(liiiclla niiomala Stiud. 


Todashiba 


Agrimonia viscidula var. japuiiica Mic]. 


Kinmizuhiki 


Phytolacca acinosa var. esculenta Max. 


Vaniagobo 


Taraxacum officinale Wigg. var. glauccscciis. 


ran]X)po 


etc. 


etc. 



Dicsc Artcn bcndtiqcn zu ihrcni Gcdcihcn cine {.jcrini^erc Feuchtig- 
kcitmcnq,c. Audi siml sic jjjcixcn extreme atmospharisclie F.inwirkunfjcn 
\venij:jcr eniprimllich. 

2. Grasartiii a)H Mittcl/inJict- 



Sciiecio krameri Vr. et Sa\. 

Polygoiiatum cuspidatuni S. ct Z. 

Thalictrum milium I., var. elatum I.ecoy. 

.Vstcr iiidicus I.. 

( >snuind.i regalis 1,. var. japonic.i .Milde. 

I'lectranthus giaucocalyx Max, var. i.ijxjuica M.i\. 

.\ster «caber Thunb. 



\'aburei;Aj-a 

lt;idori 

.\kikararval*ii 

Vonicna 

Zemmai 

Ilikioki^hi 

Shirayamagiku 



3i8 



0. Shishido: 



Patrinia scabiosacfolia I -ink. 
Patrinia villosa juss. 
Seseli livanostis var. daucifolia DC. 
Artemisia japonica Thunb. 
Cursium spicatum (Maxim.) 
Miscanthus sinensis Anders. 
Solidago virga aurca I.. 
Serratula coronata I .. 
Angelica decursiva MIcj. 
I'lcctranthus inconspicuus MU[. 
etc. 



( )minacshi 
Otokoeshi 
Ibukibofu 
Otokoyomogi 
Vamaazami 
Susuki 
AwadachisG 
Tamabdki 
Nodakc 
Vamahakka 
etc. 



^. Grasartcn iiii Thalc. 



Aralia cordata Thunb. 


Udo 


Petasites japonicus Miq. 


Fuki 


Oenanthe stolonifera DC. 


Seri 


Acorus gramineus Ait. 


Sekishd 


Astragalus sinicum I,. 


RengesG 


Iris jajionica Thunb. 


Shaga 


EquisLium arvense L. 


Sugina 


Clematis herachipolia var. Stans S. et Z. 


Kusabotan 


Vicia hirsuta Koch. 


Suzumenoendo 


Achillea sibirica Ixdeb. 


Nokogiriso 


Ranunculus acer var. japonicus Max. 


Umanoashigata 


Osmorhi/.a japonica S. et Z. 


Yabuninjin 


Rumcx japonicus Meiser. 


Gishigishi 


Angelica miqueliana Max. 


Serimodoki 


(jeranium nepalense Sweet. 


Furoso 


Carex confertidora Pott. 


Shirasuge 


Carcx gibba Wahl. 


Maskkusa 


etc. 


etc. 



\'ort;cnannte (ji'ascr sind hauptsaclilich auf alljahiiich ^^ebrannter Hara 
wahr/Ainchnicn. 



Ueljor die Einwirkiing des //rtJVf-Brenncns. 319 

Am Iku-gkammc godcihcii iiiir diejenigcii Grassarten, wclchc grossc 
Mengc von Sonnenlicht zu ilircm Wachstum bcnothigcn ; wahrend im 
Thalc jene Arten fortkommcn, wclchc zu ihrcm Gcdcihen wcnigcr Sonnen- 
licht dagcgcn mchr Fcuchtigkcit nothig habcn. Bcim Vcrgleich mit wcnig 
gebrannter Hara zcigt sich nun die auffallende Thatsache, das auch jcnc 
Grasartcn. wclchc sonst nur gcringcrc SonncnHchtmenge bcanspruchcn 
hicr cbnfalls am Kammc gefunden wcrden. Die Eigcnschaften resp. Exis- 
tenzbcdingungcn dcr Griiscr am Mittelhangcn liabcn cntsprechend ihrcr 
Lage wcnigcr ausgesprochcnc Bcsondcrhcitcn. 



III. KaI'I TKI, : W'uchsrjustandc dcr Grdscr m(f gcbraiintcr Hara. 

Auf scltcn gebrannter Hara ist die Bodcndecke dichter. sic schiitzt i\K:\\ 
Boden vor dirckter Einwirkung dcr Atmosphiirilicn und gibt in den 
Verwesungsprodukten ihrcr Pflanzcn gewissermasscn den Dungstoft' fiir die 
spiitere Generation wicder. Solchc Bodcndccken cnthalten cine grosse 
Mengc von Feuchtigkeit, u. verhindern weiterhin cine starke Austrocknung 
des Bodens durch die Sonnenstrahlcn. unter solchen giinstigen Umstiindcn 
Icben dann auch Gewiichsc, wclchc zu ihrcm Gedeihen cine gcwisse 
wenn auch miissigc Mengc von Niihrstoffen und Fcuclitigkeit unumgiingiich 
nothig haben. Auf nacktcr Hara konnen nur bestimmtc Pflanzcn lebcn. 
denen neben starkcm Sonnenlicht. hohc Wiirmc und ein sehr gcringcr 
Proccntsatz an Niihrstoffen und Feuchtigkeit nothig ist. Wenn man z. B. 
die Entwickelungsphascn von Miscanthus sinensis. Pteris aquilina, Potentilla 
fragarioides. Euphorbia sieboldiana. und Sanguisorba officinalis autmerksam 
beobachtct. so mr)gcn sic oft zwar zahlreich vorhanden. aber arm an 
Arten scin, sowie von sehr klcincm (HJcr zucrghaftem Wuchse. Ich unter- 
suchtc das Langenwachstum des .\rtuuchscs auf vcrscliiedenen Haras. 
Solchc Beobachtungcn miissen natiirlich in dcr /eit gemacht werden. wo 
die Grass-Pflanzcn ihrc T.cbensthatigkcit ebcn vollendet haben. Die 
Entwickciungszustandc dcr bctr. Gewachse sind im Einzelnen je nach- 
den Standortsvcrhiiltnisscn und Jahreszeiten ganz verschieden. So z. B. 
die Liingc-lMitwicklung von Miscanthus sinensis. Dieses hat im Spiithcrbst 



320 0. Shishido: 

des vorigcn Jahres seine Lebensthiitigkcit vollenclet und steht jetzt (am 
Ende April) absterbend auf der Hara. Ich dehntc meine Messungen 
sowohl auf die jdhrlich gebrannte Hara " Kiridoschi," als auch in auf 
" Omiyama " (wosclbst seit j JaJire nicJit gebrannt wurde) und auf 
" Nanamagari " woselbst seit lo Jaliren kein Brenner, stattfand, aus 

Kiridoschi (Nordseite) 

Liinge (das Mittel von lOo) 

Kanmi 1^-6033 m. 

Mittelhang 1-9433 ni 

Thai 2.0900 m. 

Mittel 1.8789 m. 

Omiyama (Nordseite) 
Liinge (das Mittel von 100) 



Kamm 


1. 90 1 3 m. 


Mittelhang 


2.3200 m 


Thai 


3.0833 m. 


Mittel 


2.4349 ni' 



Nanamagari (Nordseitej 

Liinge (das Mittel von lOO) 

Kamm --2135 m. 

Mittlhang 2.5013 m. 

Thai 3-4022 m. 

Mittel 2.7057 m. 

Fiir andere Grasarten konnte ich kein passendes Untersuchungs- 
material finden ; an der kleincn Untersuchung iibcr Miscanthus sinensis 
jedoch iLisst sich der ungunstige Einfluss des jiihrlichen Brennens schon 
deutlich crkennen. Richtiges, dickstengeliges Kayagras kann man nur 
auf selten irebrannter Hara antreffen. 



Uelier dio Einwirkung: dos //rtrrt-Br^nnonB. 32 1 

IV. Kapitel : Verd7idcning der BodenbcscJiaffenJicit durcli 
das Brcjmcn der Hara. 

(A) Verdnderimg der physikaliscJien EigeyiscJiaftcn des Bodens. 

Die Pflanzen bilden sovvohl als lebendc Individucn, wic auch durcli 
ihrc abgestorbcncn Korperthcilc unter den <^c\v6hnlichen Verhiiltnissen 
einc Decke, fur jede Bodenart. In diescr Decke finden dann andere 
Pflanzen wiederum ihr Gedcihen. Es iiben zweifellos verschiedene Dccken 
auf den Pflanzenwuchs und den Boden einen gunstigcn oder ungunstigen 
Einfluss je nacli dem aus. Auf viel gebranntcr //izrafliichc z. B. in 
" Kiyosumi," wo man durch das Feuer die Pflanzcn-Deckc des Bodens 
oft vcrnichtetc, iinderten sich alle Verhiiltnisse besonders die physikalischcn 
Eigenschaften des Bodens in ungiinstiger Wcise weil cr nunmehr schutzlos 
den atmospharischen Einvvirkungen direkt wiederholt ausgesetzt wurdc. 
Die flach- und mittelseit grijndigen Bodenarten finden sich in der Regel, 
entweder auf den Riickenplateaus oder an den oberen Gehangen der 
Gcbirgc, namentlich wcnn die letzteren waldlos sind ; der in den untercn 
Regionen und am Fusse der Gebirge lagerndc Boden dagegen ist mittel- 
grundig oder sogar tiefgriindig ; inbesondere bei stcilen Gebirgsgchiingen 
wird dies deutlicher erkennbar. Das kann audi auf der Hara von Kiyosumi 
nachgewiesen vverden, besonders auf der hiiufiger gebrannten Hara. 

Ich habe schon angedeutet, dass die Gebirge in Kiyosumi sclir stcil 
sind, moist iiber 35° Neigung ; wirtl nun die Pflanzen- Decke auf der Hara 
durch hiiufigcs Brennen wiederholt fast giinzlich vernichtet, so gclangt 
der fciiie gute verwittortc huniosc Boden immer melir durch die Rcgengiissc 
in grosser Menge thalwarts, so class ilie einen Riickeu bcklcidcnde Hara 
(d. h. deren Boden) sich als ausserordentlich flachgriindig u. nahrungsami 
nachweisen liisst ; nicht selten tritt sogar bald das Grundgcstein zu Tage. 
Im Thale dagegen ist durchgiingig der Boden tief- ja oft sogar schr tief- 
griindig zu finden. 

//rtnr-boden muss also umso seichtgriindiger werden, je ofter die Hara 
gebrannt wird, Auf selten gebranntcm //dtrrf-boden wird durch die Icbendcn 



-322 



0. Shishido: 



und abgcstorbenen Pflanzentheilc tier Kodcn ij^chaltcn, was in s^^rosscrer 
Tiefgrundifrkeit zum Ausdruck kommt. 



(B) Bindigkcit. 

Wie crwahnt sind im Allgemeinen die Vcrwittorungsboden dcr 
Tcrtiarperiodengcsteinc zu den fruchtbaren zu rechnen. Sobald aber, wie 
auf der Kiyosumi Hara, der Bodcniiberzug ofters abgebrannt wird, tritt 
auffallend rasch Verschlechterung ein ; alljahrlich gebrannte Hara, zeigt 
sehr lockeren oder oberfliichlich grob gekrijmelten Boden, welcher gegen 
Pflanzen- Wuchs sicli ungiinstig verhillt infolge seiner grossen Austrock- 
nung. 

Weiterhin hiingt die Empfiinglichkeit eines Bodens fiir die Erwiirmung 
durch die Sonnenstrahlen zum grossten Theile von der Beschaffenheit seiner 
Oberfliiche ab ; in dieser Beziehung gilt, dass, je grobkorniger das Gemenge 
eines Bodens ist, desto schneller und starker er sich erhitzt durch die 
Insolation, aber dass er audi um so schneller die hierdurch erhaltene 
Warmesumme wieder an die Atmosphiire abgibt, sobald die Sonne seine 
Oberfliiche nicht mehr bescheint ; je feiiikrilnicliger oder dicJitcr dagegen 
ein Boden-Gemenge sich zusammenstezt, umso langsamer erwarmt es sich, 
aber um so langer halt auch der Boden die einmal aufgenommene 
Wiirmemenge fest. Der erste Fall ist zweifellos dem Pflanzengedeihen 
scJir schiidlich, wilhrend der Ictztcrc als gijnstig bezeichnet werden muss. 
Da die selten gebrannte Hara den feingekriimelten und milden bindigen 
Boden enthiilt, so gedeihen daher auch die Pflanzen auf dieser Hara 
besser, wiihrend die jahrlich gebrannte Hara mit ihrcr schlechtcn Boden- 
struktur, nur schlechte Gras-Arten, mit extremen Anpassungsvermogen 
ein Fortkommen gestattet. 

Wasser ist ein wichtiges Lebensmittel fiir alle Pflanzen. Ohne Wasser, 
keine Pflanzen ! Fs ist also der Boden, worin die Pflanze wurzelt, nicht 
bios ihr Schiitzer gegen Winde, ihr Erwarmer in der Kiilte, sondern auch 
ihr ]Vasscr spender, wenn die Atmosphiire ihr kein Wasser darreicht ; dieses 
wichtige Amt vollbringt der l^oden einerseits durch sein Wasseransaugungs- 
\ferm(')gcn und anderseits durch seine Wasscrhaltungskraft ; das erste ist 



Ueber die Elinvirkmu? des //rira-Brennens. 323 

eine Thiitigkeit dcr Capillaritat, wclchc sicli umso gunstiger bcmcrklich 
macht, jc fcinkrumigcr der Boden ist, im zwciten Sinne wirkt am besten 
cin an Ilunuis- und Thonsubstanzcn rcichcr Kodcn. 

Den bcstcn Standort fiir die meisten Gewiichse bictcn diejenigen 
Bodenartcr,, wclche untcr den gewohnlichen Verhiiltnissen bei starker 
Wasser- Aufsaugung das in ihncn angesammelte Wasser miissig festhalten 
und miissig verdunsten. Auf der Kiyosumi Hara aber, dort wo das Gras 
jiihrlich gebrannt wird, kann der Boden diese wichtige Aufgabe nicht 
leisten, weil die Decke ja beinahe giinzlich vernichtet wird und der Boden 
direkt den atmosphiirischen Einwirkungen preisgegeben wird. In dcr 
Oberfliichenstruktur des Bodens sind grobe Kriimmel keineswegs dem 
Pflanzengedeihen zusagend. Je hiiufiger die Hara gebrannt wird, desto 
mehr trocknet der Boden aus und desto grober wird seine Struktur durch 
Auswascliung werdcn ; es herrschen Extreme von Trockniss und Fcuchtig- 
keit in solchem Boden. Auf anderer nur selten zum Brande gelangter 
Hara ist der Boden dem Pflanzenwuchsc zusagender, weil feinkorniger und 
mit grossem Wasser- Aufnahmc u. Erlialtungs- Vermogen ausgcstattet. 

(B) Vcrniindcruno- dcr organischoi Substanzcn im Boden. 

Die organischen Substanzen im Boden bilden fiir die Pflanzen niitz- 
liche ja unenthehrliche Stoffe ; man nennt dalier einen Boden, weicher 
an organischen Substanzen reich ist, einen produktiven Boden. Die 
organischen Substanzen im Boden entstelien durch hmgsame Oxydation 
abgestorbener Pflanzentheile der Ikidcndecke unter dem lunflusse dcr 
Bodenfeuchtigkeit und werden " Humus " genannt. 

Auf solcher Hara, wo das Gras alljahrlich gebrannt wird. \erniclitct 
das Feuer fast die ganze Bodendecke mit den I lumusstoffcn. wclche die 
Pflanze so nothig zum Wachsthum hat. Bei mcinen Untersuchungcn 
in Kiyosumi stiess ich demgenuiss stets auf nachthciligc \\"irkungcn des 
wiederholten /AjTcr-PreiuiL-ns. 



Ik 



324 



0. Shishido: 



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Ueber die Eiinvirkmi? des Hara-Brennens. 

II. Das Glilhen dcs gawinmcncn Bodens. 



325 



Namen. 


[ 

Fcincnle 


1 Geu icht der Feinerde 


\'ermindcrung des 








(3 gr.) nach Gluhen. 


Gewichts 










(B) 


Kiridoshi 


(Nordseite) 


3 gr. 


2.2436 gr. 


0.7564 gr. 


" 


(Siidseitc) 


3 >. 


2.2933 ,. 


0.7067 „ 


Kajisaka 


(Nordseite) 


3 „ 


2.21 iS „ 


0.7882 „ 


" 


(Siidscite) 


3 ,. 


2.199S „ 


0.8002 „ 


Omiyama 


(Nordseite) 


3 » 


2.1704 „ 


0.8296 „ 


" 


(Siidseite) 


3 ,. 


2.2388 „ 


0.7612 „ 


Musado 












' 


3 M 


2.2212 „ 


O.778S „ 


Omiyaina 












" 


3 >, 


2-1859 „ 


O.8141 „ 


Suzuriischi 














3 ,. 


2-2631 „ 


0-7369 » 


Saiiiiodai 


(A'ordscite) 


3 » 


2.2664 „ 


07336 „ 


" 


(Siidseite) 


3 » 


2-2913 ,. 


0.70S7 „ 


Suzuriischi 


(Nordseite) 


3 )> 


2.17S0 „ 


0.S220 „ 


Nananiagari 












" 


» 


2 1461 „ 


o.S>39 „ 


Musado 












' 


J » 


2-1559 ., 


0.SS41 .. 


Sene;en 














J ). 


2.1043 „ 


0.S957 ., 


„ 


(Sijtlscite) 




2.05 2S „ 

1 










0.0472 ,. 



III. Bcstimunino; dcr oro;aulscJuu Substauzcn /;;/ Bodcu. 
Aus den Tabcllcn I. und II. wird die fol-cndc : 



Xanu 



Kiridilschi (Nordseite) 

(Siidseite) 

Kajisaka (Nordseite) 

(Siidseite) 

Oiiiiyania (Nordseite) 

(Sudseite) 



Die Menj^e der 
onjf. Suhstanzeii 
sammt dem Verb 
iiiduiigswasscr in 

3 gr. Feinerde. 
(H-A) 



03790 gr. 
0.3792 .. 
0.3994 ., 
O.C3-I ,. 
0.3976 ., 
0.3056 ., 



I'roceiit der org. 

Substaiizen 

.sammt dem Ver- 

In'ndungswasscr in 

ausgctrockncter 

P'cincrdc. 



I'n^ceiu der or.;. Froccnt dor org. 



•4.45I3 % 
14-1SS9 ., 
15.2056 „ 
'4 5' 1 1 ,. 
1S.6567 .. 
14^^276 .. 



Suh.stanzen 

.sammt dem Ver- 

bindungswasscr 

in Feinerde. 



Substanzen 

sammt dem \"er- 

bindungswa s.«er 

im <.'riginalboden. 



12.6400 

'33133 

12.44(7 
16.51X0 

I mS66 



I2-S33I % 
1 2. 25 S3 

1 2.3 ;;S 

'2.1343 
15.9630 



326 




0. 


Sliishido: 






Musiido 


(Siidseite) 


0.3966 „ 


15-1501 „ 


13.2200 „ 


12.9305 „ 


Omiyama 


„ 


0.3S93 .' 


151173 » 


12.9766 „ 


12.7236 „ 


Suzuriisclii 


„ 


0.3780 „ 


14-3122 „ 


12.6000 ,, 


12.2509 „ 


Sannodai 


(Nordseite) 


0.4199 .' 


156311 „ 


13.8966 „ 


13-49S4 » 


» 


(Siidseite) 


0.3609 „ 


13-6075 -> 


12.0300 ,, 


1 1. 7641 ,, 


Suzuriichi 


(Xordseite) 


0.401 1 „ 


15-5550 V 


13-3700 „ 


13-2304 .- 


Nanamagar 


„ 


0.4180 „ 


16.3020 ,, 


13-9333 ', 


13-7954 V 


Musado 


., 


0.4453 .. 


I7.II90 ,, 


14.8433 » 


I43S17 n 


Sen gen 


„ 


0.5 1 10 „ 


19.541 1 „ 


19-0333 » 


16.8426 „ 


" 


(Siidseite) 


05069 „ 


I9.S63I „ 


16.8966 „ 


16.6040 „ 



Aus der Tabellc kann entnommen werden, dass in jahrlich gebranntcm 
Nara-hoden cine starke Verminderung der organischen Substanzen eintritt, 
wilhrend in selten <^ebranntem Hcrra-hodcn diese zunimmt. 



VIII. Absehnitt. 



Die EimvirkuuQ- dcs Hara-brcnncns in okonomischer Bcr^ieJiiing. 



In den meistcn Gegenden herrscht der Glaubc, 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, chi im letztcren Falle die Pflanzen- 
Asche nicht liinger am Ilange bleibt, sondern durch die Regengiisse 
tliahvLirts gefiihrt wirtl. Man liat eljcn nur nach bestimmtcn Artcn gesehen, 
welclie sicli unter UmstLinden auf alljalirhch gebranntcr Hara in der 
Mehrzalil fmdcn, aber niclit nach der allgcmciiioi Beschaffenheit der 
Bodendeckenvegetation und ilu'en Wuchszustiinden. Miscanthus sinensis, 
welches d^w Leuten als Dach-Deckmittel uncntbehrlich scheint, kann auf 
gebranntcr Hara zwar in Masse vorkomnicn, doch wenn man genauer 
zusieht, so fuidct man, dass der einzclne Stamm kiirzer und dunner ist, als 
auf andcrcm I'latzc, der scltcn gcbrannt wird. Diese Thatsache beweist, dass 
das l^rcnncn scinen Zwcck nur mangclliaft erfiillt, es kann soldi zwerghafte 



Uober die Ei.wirkui>i? dis y/«,-«.ll.v,n, ,,s. 327 

Miscanthus den IVdUrfn.ssc d.r Loute auf die Dauer nicht g.rccht werdcn ■ 

d.e nc t,ge„ u„d langen Ha,.c gcwinnt „.an „u. auf ™^.,,.„,„,, ^,„,; 

Auch d,e Grascr von alljahrlich gcbranntcr Hara sind vorzugswcisc "saxrc" 

Grascr und fiir das Vieh kaum seuicssbar. 

Bctrachtct man also di. Sachc u„bcfan,c„ von a„en Seitcn, so ist das 

Brcnncn der H.ra niCtswcitcr als cine scl„echtc, dure,, Nichts bcgrUnd.tc 
Gewol,nlK,t und -nn^Msck„fl,ick- .u nennen, .veil der Boden dem 
sen natUdiche,. „un,us geno„,„,e„ wird, suceessive .ur InproduktivitUt 
hc.-absu,U. In Kiyosu„,i hat n,an s,c„ in Erkenntniss dieses U.standes 
zur Aufforstung in planmassiger Weise entschlossen, da dadurcl, ein woit 
grossercn Gcwinn crzielt werdcn wird, a,s bei der bisherigcn Hara- 
WT hsebaft .elehe fa. das Land nicht die .indeste wirthschaftiiche 
Bcdeutung, hat sondcn led.glich a,s eine '-sMecJUc Gc^Muir bc.eichnet 
wcrden muss. 



XI. Abschnitt. 

Die Einioirkung dcs Brcnncns der Hara im 
laiidzvirthschaftUchen Sinne. 

Das //«™-K.enncn in Japan entstand l,a„ptsachhcl, aus den Bedurfnissen 
ckr La„dw,rthschaft. n,an woMtc dadurch cine grosse Mcnge Grundunger 
bekonnnen, alx. das Brennen der H.ra hatte wie Ubetal, so auch aufde. 
K.yosun„ //,„.„ nu,- das Gedeihen - .nUc,,.,- Grasar.cn. zur Fol^c 
wclchc m,. den Boden als D«ngn,itte, katnn «,rksan, zu nennen sind. Cuf 
jcne,- //,„.„ .,„,.„,,„, „,, ,^,,^, ,^,^ ^.,.^^^ ^ ,^^. ^^^^^_^ ^^^^^^^^^ ^^.^^^ ^^^^^ 

s.cb grosscc Mcngen von guten Species unter andcn, z. B. Legn,inose„ 

S.eKstof,«.e,ches,c aus dc- Luft ahso.-biercn, wodurch sie a,s Da„g™t.el 
sclir wirksani siiul. 

I" dc- Asehe, wcKlK. durch das Brcnnen der Bodenuberzuges erzielt 
-d. n.nden sich zwar fur die Bllanzen einige gute Xahrstofl-e.' z. B. das 
Kah: ahc,. an steilen Abhangen, wic auf der Kiyosumi //„.„ wird es 



328 0. Shishido: 

zusammcn mit den feinen produktiven Bodentheilcn durch die Regengiisse 
thalwiirts gewaschen und dadurch erst recht ein nahrungsarmer Boden 
geschaffen. Als Futtermittel sind jcnc schlccJitcn sauren Grasarten, welche 
sich auf (durch das Feuer) oft vervviisteter Hara bcfindcn, fur das Vich 
kaum verwendbar, wilhrend die auf guter, bezw. selten gebrannter Hara 
wacliseiiden Arten als ein sehr gutes Futtermittel bezeichnet werden 
miissen. Durch das Brennen der Hara werden also gute nahrungsreiche 
Grasarten vernichtct und der Boden landwirthschaftlich werthlos gemacht, 
die Hara erfiillt also gerade ihren eigentlichen Zweck umso weniger, je 
ofter sie jjebrannt wird. 



X. Absehnitt. 

Die EifrcL'irknng dt's Brcnnciis der Hara 
in forstlicJicr Hiusicht. 

I- Kapitel : Gcfalirliclikcit des Brcnncns der Hara. 

Die Forstwirthschaft bedarf im Allgemeinen grosser Vorrath-Kapitalien 
und langere Zeitriiume zu ihrer Produktion, um gute Ertriige nachhaltig 
in ihren Waldungen abzuwerfen. Unter verschiedenen Gefahren wie 
Insekten, Sturm etc, welche den Wald bedrohen, ist auch das Feuer zu 
erwilhnen. In Japan sind Waldfeuer sehr hiiufig, was hauptsachlich auf die 
achtlose Jkhandlung des Feuers zurlick zu fuhren ist, namentlich ist es das 
Brennen der Hara, welchem grosser Schaden im angrenzenden Wald 
alljiihrlich zur Last zu schreiben ist. Die Hara wird hauptsachlich im 
Friihjahre (von Februar bis Ende Miirz) gebrannt ; die an solche Hara 
angrenzenden Waldtheile stehen in grosser Gefahr. Es gibt zwar gesetzliche 
Vorschriften iiber das Brennen der Hara, wonach die Hara ohne zuvor 
erholte Erlaubniss iiberhaupt nicht gebrannt werden soil, auch muss dem 
benachbarten Grundstiick ein gewisser Schutz gegeben werden durch 
Ziehen eines sogenannten Schutzgrabens. Da diese Vorschriften nie 
beachtet werden, tritt das Feuer iiber die Schutzzone in (\k:\\ benachbarten 
Wald nur zu oft iiber. 



Ueber die Einwirkung des Ilara-Bromwns. 329 

Die Hara in Kiyosumi grenzt an ihrcr nordlichen Seite an das 
Schulwald-Arcal an, welches in Ictztes Zeit aufgeforstet wurde ; es bedarf 
deshalb eincr bcsonderen Aufsicht, vvenn man das Brennen gestatten will. 
Die Vernichtung von Wald ist eine constante Begleiterscheinung des 
Brennens der Hara und auch aus diesem Grunde mochte diese Boden- 
nutzLingsmcthode zu verwerfen sein. 

II. Kapitei. : VcrnicJitnng dcr Bodejidecke. 

Wie ich schon bemerkt habe, bildet die Pflanzen- Vegetation durch 
die Humusbildung eine Decke, welche der Erhaltung jcder Verwitterungs- 
schicht giinstig ist. An abhiingigen Lagen bildet sie einen das Fortschliim- 
men der Erdtheile verhindernden Widerstand und an sonnigen Stcllen 
schiitzt sie den Boden gegen die Sonnengluth und bewahrt so den von 
ihr bedekten Boden vor Ausdorrung, im Sommer, wic vor den Einwirkungen 
des Frostes im Winters. Sie liisst starke Regenmengen nicht direkt zum 
Boden dringen, sondern gewiihrleistet dercn allmiihlige und mehr 
gleichmassige Aufsaugung in den Boden. Das zu rasche Abfliessen des 
Wasscrs mit seinen schadlichcn Folgen wird hauptsilchlich verhindert. 
Durch das alljahrliche Brennen von //^'r^'-flachen wird nun die Humus- und 
Pflanzcndecke giinzlich vernichtct, sie kann also auch ilir wichtiges Amt 
nicht mehr vollbringen. 

III. Kapitei, : Vcrjiiindc-nitii^ dcr Na/irinij^sstoff'i- im Boden. 

Man unterschcidet zwci Artcn von Xalirungsstoffcn im Boden namlich 
die organischcn wnd anorganischcn oilcr mincralischen Stofle. Die org- 
anischcn Substan/.cn im Ix^clen enstchen durch den \*er\vesungsproccss der 
abgestorbenen Organismen (d. h. rOanzcn uwd rhiore\ Auf Ham und 
Waldboden sind sie meist von abgestorbenen Pflanzentheiien gebildct. 

Der Boden enthiilt cine um so grossere Mcnge von Humussubstanzcn. 
jc grosser die Zahl tier tlort lebenden und absterbenden Pflanzen 
ist ; auf Hara aber, welche alljiUirlich gebrannt wird, gibt cs nur cine 
geringe Menge abgestorbencr Ptlanzen ; auf der schutzloscn Hara bringen 



330 0. Shishido: 

die Regengiisse etc. unaufhorlich Humussubstanzen, feinen Sand und erdigc 
Theile von den Berggebiingen berab in die Tbalcr zur Ablagerung oder 
in die Biicbe zum wciteren Transport ; wenn man also an die Aufforstung 
oft gebrannter Hara gebt, so feblen dem Boden jene Nabrungsstoffe, welcbe 
zum Gedeiben der jungen Bestiinde so notwendig sind ; man kann daber 
auf solcben scblechten Boden aucb nur jene Arten von Biiumen pflanzen, 
welcbe die geringsten Anspriicbe an die Niibrkraft des Bodens stellen, auf 
die besseren Baumarten muss man also von vorn weg verzicbten, was 
einem erheblicben wirtscbaftlicben Nacbtbcil gleicbkommt. Auf die 
traurigen Folgen der Gescbiebefubrung der Biicbe und Fliisse infolge der 
//(?/'<?-bedeckung der Gebirge sie bier ]iur kurz bingewiesen. 

IV. Kapitel : TrockcnJieit des Bodens. 

Wo dem Boden die notbige Menge von Wasser febit, da bleiben die 
Nabrungssubstanzen in ungeloster Form, was wiederum Nabrungsnotb fur 
die Pflanzen bedeutet ; aucb konnen die Pflanzen ibre zu ibrem Gedeiben 
notbige Transpirationsgrosse bei Wassermangel nicbt aufrecbt ertalten u. 
sterben ab. 1st nun der Boden mit Pflanzen und Humusdecke gescbiitzt, so 
kann er eine bedeutende Wassermenge aufnehmen u. festhalten, welcbe 
langsam aber sicber u. wirksam den Pflanzen zu Gute kommt, wabrend 
auf nackten Hard s die meteorischen Niederschliige oberfliicblicb rascb 
abfliessen und der Boden der Trockenbeit ausliefern. Trockenbeit des 
Bodens aber ist fiir die Forstwirtscbaft oft ein sebr grosses Hinderniss ; gute 
Bestiinde konnen nur auf jenen Boden gedeiben, wo ibnen eine gewisse 
Menge Feucbtigkeit nicbt feblt. 

Nacbdem die Hara in Kiyosumi, alljiibrlicb gebrannt wird, bat sie audi 
nur eine scbwacbe Pflanzen- & Humusdecke. Aus diesem Grunde kommt 
clann der fast nackte Boden durcb I^inwirkung der Atmospbiire, namentlich 
durcb die starke Besonnung mebr u. mebr berab. Wollte man solcben Boden 
sofort aufforsten, so wiirden die Pflanzen tlie notbige Wasser- und Nabrungs- 
menge im Boden kaum fmckn und zu Grunde geben. Dagegen bat man 
stets leicbteres Spiel mit jener Hara, welcbe seltener gebrannt ist, sie 
setzt der .\urrorstun</ viel weniger Scbwierigkeiten entge<a:n. 



rdnT (lie Kiiiwirkiiiijr <I<'s //(^/rrf-'irciiiiois. 331 



Schluss. 

Alls (Icii hisiK r niitL;cthciltcn l)cob;ichtLini^cii niai^ crsclien ucrdcn, dass 
die Kiyosumi Ilnra, da sic in dcr subtropisclicii Zone mit dcrcn t^rosser 
Mcngc an Luftfcuchtif^keit bei seltencm Frost und Schnec Hcgt, in Bczui^ auf 
das Pflanzeni^edeihen zu den Li'unstiq'cn Pliitzcn t^crechnet werdcn muss. 
Trotzdcm wird zur Zeit von den <;rossen Fliichcn der Kiyosumi Hara 
in ihrcr " Ilara " — l^^orm, nur ij^eringer Nutzcn f^^ezogen, sic sind dc facto 
Nichts weiter als unproduktive Flachcn. Oftmaligcs l^rcnncn dcr Hara 
vcrursacht fo]f,^ende Nachthcilc : 

1. Die Pflanzcn, Eodcndccke ist durch das hiiufige l^rcnncn dcr Hara 
fast L^anz zuni Vcrscliwindcn t^cbracht. 

2. dcr J^odcn wird tlirekt den Atmospliarilicn grcisgcgcben. 

3. die Regenwasscr fliesscn obcrflilchlich ab und vcrursachcn kcinc 
Zunahmc der Bodenfeuchtigkcit. 

4. tier Bodcn gcht \\\ grobc Kriimnielstruktur iiber. 

5. tlie produktive Hcnlcnkrunmie wird durcli die Rcgcngiisse thalwiirts 
abgefiihrt. 

6. 13ic Bodcnverwitterunsgschicht wird dadurch seichtcr und seichtcr. 

7. ICs tritt im Laufe der Zeit ITumus-Vcrarmung ini l^oden ein. 

S. die Lichtgrilser ki^mmen auf \'icl gebrannter ffara in Ubcrzalil 

vor. 
y. die (irasspecies werdcn durch das Brenncn in ilircr Zahl \crniindcrt. 

10. die Gcwachse auf oftnial gebrannter Hara ncigcn zur Dcgcncrirung. 

11. Hauniarten xcrschw intlcn soliald cine /Ar/v? c")fter gebrannt win!. 

12. wenn cine Ilara li'ingcrc Zeit niclit gehrannt winl. so geht sic wicdcr 
in W'ald i'lbir, zucrst erschcint " (}uercus gkimlulifcra " in liicsigcr 
Zone wieder. 

13. (his I lauptprodukt dcr Hara, i^d. I1. Kaya) Liras. wird liurch oftmaligcs 
Hrcnncn wcdcr in richtigcr Bonitlit noch ^Icngc crzcugt. 

14. die Aschc, wclchc durch ikis l^renncn ilcr (Eraser cntstcht. kann 
auf dcn\ gcncigtcn /Ar;vr-bo(.lcn nicht hcgcn bleibcn, sic hat also als 
Diingcr aucli kcincn W'crth. 



332 0. Shisliiilo: 

15. das Hara-hrcnncn ist in cler Niihc von Waldungen am gcfiihrlichstcn. 

16. die Wachsthumsenergie der Griiscr sinkt durch oftes Brennen herab. 
Die Hoffnung, durch Brennen Bodcn in seiner Produktivitiit zu heben, 

ist im Gebirge wenigstens (wie in Kiyosumi Hara) eine irrige. Die 
japanische Hara, welche die Haupterscheinungsform der Bodenbenutzung 
im Berglande ist, veriindert den Boden in nachtheiliger Weisc, fiihrt 
Verarmung des ungeschiitzten Bodens durch Auswaschung und Verhagerung 
iiberall herbei, ja nicht selten tritt das nakte Grundgestcin auf japanischcr 
Hara zu Tage. 

Ich schHesse meine Beobachtungcn damit, dass ich behaupte, dass das 
Hara-hxcww^n V'Om Stand punkt der Landwirthschaft wie, der Forstwirth- 
schaft betrachtet, einen entschieden nachtheihgen Einfluss auf die Hara 
sclbst ausiibt, ja dass es geradezu den Hara-ho^cw der Verodung preisgibt. 

Es wiiren nach meiner Ansicht die meisten Hara in Japan, weil 
sie als Hara nicht nur oJluc wirthschaftHche Bedeutung sind, sondern 
geradezu landverwiistend wirken, alsbald aufzuforsten. Wo dies nicht 
moghch ist, konnen gute Griiser nur nach Authoren jedes Brennens erwartet 
werden. 



INHALT. 



Einleitung 

I Abschnitt 

Die Gcschiclitc dcr " Kiyosumi llara." 

II. Abschnitt 
Zustand dcr A7;v;i' ;/;/// Hara. 

I. Kapitcl : l^odenzustilnde. 

(A) Lai^c und Flachc. ... 

(B) Bodcn (Grund _G^estcinc). 

II. Kapitel : Das Klima 

III. Abschnitt 

Cultur dcr (jcjj^cnd bci Kiyosumi. 

IV. Abschnitt 

Die Hara und ilirc l^csitzvcrhiiltnissc 

I. Kapitcl: Bcsitzarten. ... 

II. Kapitcl: (Tcwinnun<^ dcs Haraprodukte 

V. Abschnitt 

Das Z)/r////<:v/ tier llara 

I. Kapitcl: Zwcck dcs Brcnncns 

(A) lOntstchunt^- aus alter Sittc 

(B) Landwirthschaftlichc Bcni'itzuni^ dcr Mara. 

(C) Werth ilcs Harabrcnncns zur Gcwinnun^" von Fiitt^r- 

t^rasern 

II. Kapitcl: X'crfahrcn dcs 1 larabrcnncns 

III. Kapitcl: Zcit dcs I larabrcnncns 

IV. Kapitcl: Die Xfcthotlc ilcs I larabrcnncns 

VI. Abschnitt 

Kinfluss auf physik.dischc l-atj;enschaften tics 7)\'f/tv/jr 

I. Kapitel : i^a) Ticfc, (b) Bindigkeit, {c) Fcuchtigkeit. 

II. Kapitel: .\usscrc Zustiindc dcr Mara 

III. Ka])itcl : Das V^crhiiltniss zwischen Bodcn und Klima 

(Verw itterunsjsprocess.) 



lAGE. 

26S. 

269. 
269. 
269. 
270. 
270. 



-/ 0- 
-73- 

274. 

-74- 
274. 
274. 



-7S- 

-7S- 
-7'-- 
-77- 

2 78. 



134 



liihalt. 



VII. Abschnitt 

Vergleich (k-r in \'crschicdciicni Hrcnn-Turnus gcbraiiiitcn Ilara- 

Pliitzc 279. 

I. Kapitel : Untcrsuchung-cn ini Einzcliicn, Probcfliichcn. ..280. 

n. Kapitel: T'olgcn dcs Brcnncns dcr Hara 286. 

(A) V^craJidcniUi^ dcr Gcii'aclLsartcn. 286. 

1. Vcrmindcrung dcr Gcwachsartcn 315'. 

2. Untcr dcni lircnnen nicht Icidcndc (icwaclis- 
Arten 316. 

(Z>) Grassartcii dcr vcrscliicdcjicii Standortc 317. 

1. Grasartcn auf don Riickcn dcr Bcrgc :;i 

2. Grasartcn am MittclhaiiQc. 

3. Grasartcn im Thalc 

III. Kapitel : Wuchszustilndc dcr Griiscr auf gcbranntcr 

Hara 319 

1\'. Kapital : VcrLlndcrung dcr Podcnbcschaffcnhcit durch 

das Brenncn dcr Hara 321 

(A) VerandcrunQ- dcr physikalischcn Kigcnschaftcn des 
Bodcns. ... 321 

(B) Vcrmindcrung dcr organischcn Substanzcn ini Bodcn. 



317- 
318. 



VIII. Abschnitt 

Die Einwirkung dcs 1 larabrcnncns in (')konomischcr Bczichung. .. 326. 

IX. Abschnitt 

Die Einwirknng dcs Brcniicns dcr Hara in laiulwirthsch.iftliclKin 

^i>i»^ 32;- 

X. Abschnitt 

Die I^inwirkung dcs J^rcnnens dcr ] lara \\\ forstUcJicr Hinsicht. .. 328. 



I. Kapitel: (iefiihrlichkcit des Brenncns dcr Hara. 

II. Kapitel: Vcrnichtung tier Bodendccke 

III. Kapitel : Verminderung dcr NahrungsstolTc in JmhIcii. 
I\'. Ka})itcl : Trockenhcit des Bodens 

Schiuss. 



• 328. 

• 329- 
329. 

•• 33v 



^o. 



Die zLikiinftige Bewirtschaftungsform des japanischen Wa'des! 

vox 

Dr. Hefele. 

Ki^'l. Iniyr. I'lnslnicisier u. Pro/cssor in 1 ukv>. 



Die Umwill/Ainc;" ties Jahres t868 ist audi \'on c,M'ossem Minflusse auf die 
japanischen \\ alduni^en c^ewescn. Walirend friiher wolil die mcisten dcr 
Feudalherren die ihnen iintcrstellten Kronforste untl Staatswaldungcn 
wenii^stens in einiLjerniassin j^utem Zustande erhielten, wenn auch cine 
bcsondere W'aldwirtschaft sicli nicht entfaltete, so ist doch in den letzten 
Jahrzehnten nianclies Stuck treffiichen Staatswaldes der Eec^ehrlichkeit 
insbcsonders bilucrlicher Kreise und den scliwankcndcn. wecliselnden 
Auschauunfj^en iiber die Lr)sunq" der volkswirtschaftlichen Fragen nach der 
Restauration in Japan zuni Opfer gefallen. Ausscrdem besass die neue 
Centralgcwalt \-ielfacli weder die Macht nocli die Organe, ihre Autoritiit 
o-eniigcnd geltend zu maclien, uni tlen durch SilkularisiruuL^ von Klostcr- 
giitern, Kinverleibung' xon (^lemeindsforsten etc. \ernielirten Staatswaldbesitr. 
vor I^'orstfrevei zu schi'ilzen. 

Audi der rri\-at\\aidioesitz unterlag- nianigfaclien \'eranderunL,fcn. 
Armuth und Xersdiuldung \ieler (irund-Besitzcr, welclie bei dor 
Umgcstaltung- der allgenieinen \'erliiUtnisse, sicli der \euzeit nicht 
anzupassen xerniochten, fiihrte hiuifig zuni Wrkauf, zur Zerstuckclung" 
mul X'ernichtung \-on W'aldern. XanientHch haben aber Theihuigen voii 
W'aKlgrund unter ch'e eheileni nur zu Nutzungeii lierechtigten am vorher 
gcnieinsanien W'aKle i(,ienieinde\\ akle zur x'orauszusehenden sicheren 
W'rnichtung, Dexastirung <uler wenigstens X'ersclilechterung der 'I heilstiicke 
gefiihrt. Auch will niidi betliinken. class tlie hier auf dein Tapier so gcordnct 
und genau festgestellte C^beraufsicht iles Staates viber die Cienieindewaldungon 
in }">raxi nicht gcniigend strenge zur Anwendung komnit. Dor factischc 
Zustand iler nieisten bauerhchen rTenieindewaldungon spricht hicrflir. 



336 Dr. Hefelc: 

Habsucht and mangclndes Verstandniss veranlassen ferner gar manchc 
kleinc Privatwaldbesitzer zu einer Nutzungsweise, welchc in vielcn Fallen 
direct der Zerstorung gleichkommt. Die Eigenthumer grossercr Privat- 
waldungen sind bis zu einem gewissen Grade indessen bcreits zum tieferen 
Vcrstiindnisse des Werthes und der Bedcutung des Waldes gelangt und 
wirthschaften im wohlverstandenen eigenen Interesse etwas mehr 
conservativ'. Am besten uurde der Wald wohl da conservirt, wo die 
fehlende Aufschliessung der Gegend und die geringe Revolkerungsziffer 
ihn von selbst bis heute geschiitzt haben und es gilt dies nicht bios fiir den 
Privat-, sondern audi fur den Staatswald. Es mochte fraglich erscheinen ob 
nicht finanzielle Schwierigkeiten des Staates den dort vorhandenen reichen 
Vorriithen ein schnelles Todesurtheil gesprochen batten, wilre ihre Lage 
zum Verkehr nur ein bischen giinstiger gewesen. 

Darin beruht auch heute noch die Gefahr, dass das Bestreben hohe 
Ertriige umjedcn Prcis aus einem Walde herauszuholen, die heiligste Pflicht 
gegen das nationale Wohl vergessen lasst, nemlich die Sorge fur die 
ungeschmiilerte Uberlieferung eines werthvollen Waldbestandes an die 
Nachwelt durch entsprechende Form der Nutzung & Verjiingung. Liegt 
ja doch der Werth der Wiilder nicht bios in ihreni Rentenertrag ; die 
Uberschwemmungen und Wasserkatastrophen, welche der Vernichtung des 
Bergvvaldes z. B. allenthalben zu folgen pflegen, reden eine zu deutliche 
Sprache, als dass sie misszuverstehen wilre. Wohl sind Gesetze zum Schutze 
soldier Forstc erlassen, aber ein Blick in die realc Wirklichkeit zeigt zur 
Geniige, dass das Wollen dem Konnen noch nicht die Wagschale 
halt. 

Der Staatswald leidet abgesehen von einigen grosseren Complexen auch 
nicht wenig durch die Zersplitterung seines Areales ; es wird eine stete 
Sorge der Regierung sein miissen, eine Verbindung der zerstreuten Theile 
auf dem Wege des Ankaufes odcr Austausches herzustellcn. Wenn man 
sicli mit Recht zur Abstossung einiger selir abseits liegender Kleinflilchen 
entschliesst, so sollte doch Verschleuderung unter alien Umstanden 
vcrmicden werden. 

Eine intensivere Pflege muss zweifellos durch den ncithigen gesetz- 
miissicren Druck der Gcuicivdcivald ini ^etheilten und unLfetlieilten Besitze 



Die ;;iikiinftigo Bewirtschaftiingsform des jiipanischen Waldes! 337 

crfahrcn. Auf wclchc abschussigcr Ebcnc man sich hicrin momcntau 
befindet, zcifrt jcdc Excursion durch das Land. 

Das Bcispiel, welches der Staat den Privaten durcli seine eigene 
Wirtschaft zu t^eben bcrufen ist, setzt aber voraus, dass er sich selbst iiber 
seine cinzuschlagcnde Richtung klar sei und mit alien Mitteln auch Ernst 
machcn will, sowohl (lurch die nothigc durchgreifendc Reformirung seiner 
Verwaltung als auch durch consequente Befolgung gcwisser Wirtschaft- 
gru7idsatze. Die wichtigste Frage ist hiehei zweifellos jene nach der 
zwcckmiissigsten Art der Nutzung und damit zusammenhangend die 
Untersuchung der den Verhiiltnissen am besten gerccJit ivcrdcndcn Vcr- 
jilngungs-und Bestandsforin. In Ubung sind in Japan in der Hauptsache 
zur Zeit zwei Hauptnutzungsarten, einmal KaJilscJilag auf grosserer Fliiche, 
und dann eine Art von auszugsweiser Nutzung, welche falsclilicJi den 
Namen cines Plcntcrbctricbs erhalten hat. 

Dem Kahlschlag auf grosser zusammenhangender Fliiche musste friihcr 
da, wo durch einen Wass?rlauf die Waldungen giinstige Abfuhrbedingungcn 
fur ihre Schlagergebnisse hatten, eine gewisse Berechtigung zugcstanden 
warden, immerhin sind die aus ihm hervorgehenden Nachthcile vom 
waldbaulichen Standpunkte so bedeutend, dass man von ihm ubcrall, wo 
es nur irgend moglich ist, abzukommen trachten wird. Nicht bios sind 
alle Gefahren wic Frost, Diirrc, Unkraut (Bambus) im crhohten Masse 
bei dieser Verjiingungsform gegeben, sondern die nachgczogcnen Bestiinde 
nahezu gleichen Alters auf Flachen von grosser Ausdehnung bilden auch 
einmal spiiter fiir den Wirtschafter sehr schwierig zu realisirendc Hiebsob- 
jektc ; umso schwieriger je weniger ctwa die Aufschlicssung der jetzigen 
grossen Verjiingungsschlage solcher Pliitze bis zur seincrzeitigen Haubarkeit 
dcrselben fortgeschritten sein sollte. 

Man d.irf nicht \-ergessen, dass cs nicht Aufgabe einor Wirtschaft sein 
kann, nur IWild schlcchtJiin nachzuziehen, sondern die Gcgenwart hat die 
heiligste Verpflichtung, dom Stande dcs Wissens entsprechentl. auch 
tecJniisch vollkonimcnc Bestandsbilder der Nachwolt zu iiberliefern. 

Die Gewissensberuhigung, welche viclHich ilarin gcsucht wird, dass 
man die grossen Schliige mit Nadelholz in irgend cinem X'crbandc durch 
Pflanzung in Bestockung brinq-t, ist in viclcn Fallen cin grosses Armuths- 



330 Dr. Hofelo: 

7,cugniss fehlciidcr Ubcrlc^un^;" unci Klarhcit i'lbcr die (.irundzuL^e cincr 
geordneten Wirtschaft. 

Zicllosc Auftbrstung" L^ibt wohl Wald, ob abcr don nach Betrachtuni;' 
allcr Verhaltnissc richtigoi Wald. darnach frai^t man, wic niir auf Grund 
niciner cic^cnen Anschaiiung durch Reisen zwcifellos isc, \'iel zu wenig. 
Derii ki'instliclicn Kulturwald glcichoi Alters abcr haftct ein grosser 
Naclitheil an ; man hat wenigstens in Europa diese Erfahrung aus 
bittercn Verlusten gesammelt, dass nemlich die Insektencalamitiiten in 
grosserem Umfange und verderblicher aufzutreten pflegen, als sonst. 

Japan ist zwar in der glucklichen Lage von solchen Catastrophen bisher 
verschont zu sein, ob abcr die unter dem Drucke der wirtschaftlichen 
Notwendigkeit eintretende Umgestaltung seiner Waldungen nach innerer 
Verfassung und nach Holzarten niclit dieselben mit sich bringen werde, 
dafiir fehlt vorderhand jede Erfahrung. Wie erwahnt wiirde man gut daran 
thun, die A'loglichkeit zu bcriicksichtigen. Auch die Verlegung des 
Schwergewichtes nur nach cincr Holzart hin, z. B. der Cryptomcrie, hat. wenn 
zu uniform zur Geltung gcbracht, manche recht bedenkliche Seite. ^lan 
solltc stets darnacli streben ]\Iisch-bestande zu erzichen, um bei einer ev. 
Anderung der Conjunktur des Absatzes nicht auf das Trockene gesetzt zu 
sein und eine bessere. intensivere Bodenausnutzung zu erzielen. Das 
stellenweise bemcrkbare g'knrjlicJic V^erdriingen der Laubwaldungen durch 
Nadclholzer ist ein spiiter schwer Avieder gut zu machender Fehler. 

Zweifellos ist ja der Nadelwald in Japan iiberwiegend werthvoller und 
man winl deshalb Nichts dagegen einzuwendcn habcn, wenn der Laubwald 
l)raktisch in (\.\<^ Inferioritilt getlrilngt wird.aber ihn ganz vom Zukunftswalde 
auszuschliessen. \\\c man das an verschiedenen I^lLitzen bereits bemerken 
kann, ist ebenfalls verfehlt. Ich glanbc man gibt ihm am besten die Rolle 
eines Zwischen-und Unterstandsgliedes. dadurch wird die Bodenthiitigkcit 
besser gewahrt als tlurch die reinen Nadelholzbestiuide. Der rcinc Laub- 
wald, wo er allein angiingig erscheint, wird zweifellos im hochwaldartigen 
Mittelwalde stine beste Wirtschaftsform haben. 

Auf geeignetem Bodcn und bei entsprechenden sonstigen V'erhaltnissen 
ist audi siclur der Laubwakl noch rentabel. sobald nur auf l'>ziehung eines 
sehr wertlivolleii Materiales hingearbeitet wird. Schlechte Bcklen sind fiir 



Die znkiiiiftjgc Bewirtschaftcingsforui des japanischon Waldos! 339 

Laubwald als Hauptnutzholzwald aufzugeben, cla hier die crwiihnten 
Voraussctzungen hochwcrtigcr Produktion nicht vorhanden sind. 

Die grosse Ausdchnung dcr Flache nach, nimmt in Zukunft dcr 
Laubwald in Japan sichcr nicht mchr cin. und mit Rccht, denn cine bessere 
Wirtschaft sucht Rcntabilitixt, aber audi waldbaulichc Gcsichtspunktc 
insbcsondcrc Vielseitigkeit und Nachhaltigkeit mchr in Einklang zu bringen, 
das fiihrt dann unmittclbar zu gcmiscJiteii Waldungen. Fiir gcmischten 
Wald ist abcr die Grosskahlflachenverjiingung die ungeeignctstc und muss 
ihr auch aus dieseni Grande der Wertli, den sic in der Vergangenheit 
hatte, volhg abgesprochen wcrden. 

Ebenso unpassend ist aber dcr grossc Kahlschlag, dcr im Wesentlichen 
seine Heimath in der Ebene hatte, fiir gcbirgigc Lander, wo eben die 
Gefiihrdung des Terranis durch die meteorischen Xiederschlage in ganz 
anderer Art sich iiussert. 

So viele verodete Pliitze des Berglandes, welche hcute in ihrem 
zerrissencn, den Abschwemmungswassern preisgegebenen Boden. die 
Zufuhrquelle der Geschiebe zu den verderbHchen Wildbiichen und Flussen 
darstellen, sind sichcr grossenthcils nur auf Kahlschlagwirtschaft im grossen 
Stile zuriickzufuhrcn. \\'ir batten gewiss noch viel traurigere Bilder der 
Verwiistung des l-5ergtcrrains in Japan vor uns und cine noch viel drohendere 
Gefahr tier Wildwasscr, wciui nicht die beispiellose Reproduktionskraft des 
Podens mit ihrcr unendlich manigfaltigen rflanzendecke hier zu Hilfe kiime. 
Dass man abcr daniit auf (.lie Dauer nicht auskommt. das beweisst zur 
Geniige cine Augcnschcinnahme im (icbirge und die flochwasserschadcn- 
statistik. Die l^xtrcmc bcriihrcn sich untl nach dicsem Cirundsatze scheint 
man auch in Japan xorgcluii zu wollcn, (.Iciui naclulem zwcifellos die 
Fehlcr tier altcn bis hcute im Schwung befindlichcn Cirosskahhchlag-damit 
kunstlic/ii-n Vcrjiingidigs- sou'ic Aus:;iigs/itiintngs- Tolitik sich nicht mchr 
laugncn lasscn, will man sich zum tlircktcn Gegcnthcil bckchrcn, namlicii 
zur )iaturlichcn ]iorst-\\\\(\ gnippcnicciscn Wrjiingung. 

Die Nutzung dcr Altholzbcstilnde in langcn Verjungungszcitraumcn und 
die Nachzucht dcr ncucn Generation auf naturlichcm Wcgc in Gruppen und 
Horstcn. untcr Zuhilfcnahmc stcUcnwciscn kiinstlichcn Anbaucs. entspricht 
ja auch dcr IVwcgungsrichtung wclchc die modcntc waldbaulichc 



340 »r. Hefelo: 

Kntwicklung in Europa cingcschlagcn hat, somit cin Grund niclir, so mcint 
man hier, sofort in radikalstcr Weise vorzugchcn und zwei Treffer auf einmal 
zu erzielen, niimlich 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 begcgnen und die Verfechter derselben 
fuhren als Argument an, dass diesc Form der Bestandsnutzung und Ver- 
jiingung nicht einmal ncii sei, sondern lange Jticr zu Lande geiibt. Ws 
Beweis wird dann eine vorhandene sogenannte Plenterwirtschaft theoretisch 
und bei Gelegenheit auch in praxi vorgczcigt. 

Wenn man eine rcgcllosc Ausraubung des werthvollsten Materiales aus 
alten BestLinden und eine volligc Gleichgultigkeit iiber die inncre Beschaffen- 
heit und den ivirtJiscJiaftUcJicn Werth der Nachzucht, eine Plenterwirtschaft 
heisst, dann allerdings liabcn die Anhilnger dieser Richtung Recht. Der 
Ilinweis auf die Natur, welche ja auch in ahnlicher Weisc, wenn sic durch 
die Hand des Menschen nicht gestort wird, den Wald regenerirt hat, ist 
umso weniger gerechtfertigt, als sic cben den Wald niir als Sclhstzivcck 
weiter produzirt und verjiingt, olinc sich um Zeit und Werth zu kummcrn. 

Die Fragc nacli Zeit ?i. ]]'crt/i ist aber das Grundelemcnt einer 
heutigen W^aldwirtschaft, in der Beriicksichtigung dieser Faktoren liegt das 
Characteristicum des Begriffes " Wirtschaft." Eines der Hauptziele einer 
ivirklicJi okonomischen Betriebes muss die Erreichung des besten Effektes 
untcr Aufwand gcrmgster Mittel sein. 

Das tcc/inisch Vollkovimcnstc nach den moglichen Verhiiltnissen zu 
leisten, iiicJit aber bios iibcrliaupt Production zu treibcn, ist Aufgabe der 
modernen P'orstmannes. XaturlicJic horst-und gruppenweise Verjmigung, 
{FeJimelscJiagforvi) imd bci Aiisdilnuiug des Verjungszeitrannics mif den 
ganzen Umtrieb {Feviel oder Plentcrforni) kann das Vorliaiidenscin cbies 
guteji mid e7itivicklungsfaJngcii Verwuchses oder die ScJiaffung schoner guter 
V^erjiingungsgruppcn nicht entbehren. Wo sic kritiklos j'eden natiirlichen 
Vorwuchs beniitzt, auch wenn er mit 50 oder 70 Jahren hochstens i meter 
hoch ist, da verdient ein solches Verfalucn den Namen einer Wirtschaft 
iiberhaupt nicht mehr, es wird blanker Raubbau, dessen blendcn-soUende 
geringe Kultur-Kosten nichts weiter als blanke Tiiuschung Unerfahrener 
bezwecken. 



Die znkiiiiftif^e ficwirtsoliaftmisrsforiii dcs japaiiisohoii Waldos! 34 ' 

Glaubt man sich abcr damit bcruhij^cn zu konncn, class aus solchcn 
schlcclitcii, obcn crwilhntcn Vorwiichsen ein gcsundcr geschlossener 
Wirtschaftswald sich entwickeln wcrdc, so wiirdc das die Wahrheit des 
Satzes voraiissctzcn, dass cine Jahrzclintelan|:^ nntcrdri'ickte Baumpflanzc bci 
(lewahrung von entsprechcndem Lichtgcnuss und Kroncnfrcihcit sich ebenso 
zum Hauptstamm cntwickehi konne, wie cine gcsundc, durch Druck nicJit 
degenerirtc. Wie grundfalsch cine solche Annahmc ist, zeigt schon das 
Fiasco des auf idinlichcn Voraussctzungen basierenden Rorggrcve'schen 
Durchforstungsverfahrens. Vollends in den JidJicj'cii Rcgionen der Berge 
ist ein soldier h'chmclschlag oder Plcnterbetricb zwccklos. ]"/V/ Material 
kann nicht gewonnen werden und wo man nur mehr einzelne Stiimme 
nutzen kann, da ist man meines Erachtcns in die Region des 
ScJLiitzzvaldcs eingetreten oder ihr bedenklich nahc gekommen und da 
muss, so lange die Waldungen der tieferen Lageii noch keiner richtigen 
Wirtschaft unterstellt sind. einfach vorerst die Hand davon gelassen 
werden. 

Man i'lbersicht aber audi an dcii Platzen, wo aus natiirlichen Griinden 
liorst und ij^ntppcnivcisc Natur- X'erjungung moglich wiire, ein paar Haupt- 
bcdmgiingcn einer solchcn. Wirtschaft vol/konimcii. Sic setzt nilmlich. wenn 
sie riclitig betrieben werden will, cinmal cine \ccitgcJicndc Aufschliessung 
der Waldungen durch Wege etc. voraus und kann ferner auch nur von 
einem erfahrenen und gcscliultoi Personal exccutiert werden. 

Ob diese conditionrs si//i- qua iioii grundlegender Xatur fur Japan 
zutreffen oder nicht. dariiher glaulie icli besser kcin Wort \erlieren zu 
sollen. Dinge, w-elche weitschaueiulen l^Iick. I'msicht und praktische Schu- 
lung der technisch herangebildcten Peamten sowie cine vorziigliche Dressur 
iler Vollzugsorgane erfordern, konncn meines Krachtens nicht wie cine 
Maschine einfach gekauft und in Gang gesctzt werden und im Decreticren 
ist man zweifellos weniger becngt. als in der Durchfiihrung in praxi. Die 
ungchciirc Verantwortung aber. die darin liegt. dass man ein Xationalgut. 
wie lien Wald. der. bei richtiger Hehaiullung, eine der besten Kinnahm- 
(luellen tics Staates sein kann \\\\\\ in andern Landern auch ist. zu 
gcivagtcn lixpcrimcntcn mit zweifellos sclilcchton .\usgang beniitzt. gibt 
ilodi W(>hl .inch zu dcnken 



342 Dr. Hefele: 

Die gaiisc Frage der Rciitahilitat dcs Waldcs in Japan ist cine Fragc 
der Entiuicklung der Coninuinicationslinien ! 

Welche Nutzung und Verjiingungsform ware demnach empfehlenswerth, 
bis dieser Entwicklung des Verkehrs durch Wege etc. mehr Rechung 
getragen ist ? 

Zweifellos die Kahlschlagsfonn in rclativ schnialen Se/ddgen, im 
Bergterrain der Hangrichtung folgend, also Sannisch/ag mit nackfolgendcr 
ki'instlicJier ]^crjiingung durcJi Saat oder Pflanzung. 

Damit vermeiden wir eincrseits die Gefahren des Grosskahlschlages, und 
bekommen mcJir Material auf jedem gegebenen Hiebsplatze, als wie dies 
bei richtiger Fehmelschlag oder Fehmelwirtschaft moglich ist; daj'anf vauss 
aber gesehen uerden, da eben die geringe Ausdehnung des Wegenetzes 
fur manche substituirend eintretende Bringungsmethoden einen JiinreicJicnden 
Materialanfall als Bedingung fiir die Rentabilitiit der Bringungsanlagc 
crfordert. Die Saumkahlschlagform ist ferner technisch einfach und also 
mit dem vorhandencn Personale eher durchfiihrbar, sie ermoglicht die 
Bringung iiber die Schlagflache selbst in ungenirter Weise, und ist z. B. die 
giinstigste Abholzungsform in Bergliindern, denen gute Hangaufschliessung 
durch Wege mangelt. Unter Zuhilfenahme entsprechenden Hiebswechsels 
kann audi die sichere Nachzucht von Bestiinden jeder Holzart oder 
Mischung div. Holzarten sehr gut ausgefiihrt werden. 

Das m. E. nach den dernialigcn Verhilltnissen auf lange Zeit hinaiis 
in Japan Rrreiclibarc ist damit gekennzeichnet. Wird einmal ein wohl- 
iiberlegtes richtiges Fundament, ein solider Unterbau, fiir die weitere 
r>nt\vicklung durcli Reorganisation der Waldwirtschaft Grundsiitze mit 
zielbewustem Krsat?^ der ubersturtsien, iveder gc7ii(gend gckamite7i, nock in 
Hirer Wirkung geiiiigend gescJiatztcn MassnaJimc7i gegeben, dann wird audi 
die Zeit sicli nilhern, wo der primiiren Fundanientalaiifgabc, der Anf- 
scJilicssnng der IValdtuigen durch Wege etc., o/ine Schwierigkeit eine 
natiirliclie Verjiingungsform europ. Stils wird folgen konnen. 

Teehniker, welche se/bst praetiseJie Wirtscliafter sind, miisscn aber 
erst das Personal fiir diesc hohe Auforderung mittlerweile sehulen, was 
bei localer langsamer Uberleitung (der Aufschliesung entsprcchend) vom 
Sannise/i/ag zur fenielse/i/ag'-u'eisen I'^er/fingnng luid s/wj's/ an kleiiien 



m znkiinftige Bewirtscliaftunsrsform des jaiianisclicn WaWes ! 343 

Objecten probirt, bci consequenter r/././.;^.. Methods auch crrciclu werden 

kann. 

Schwierigk-eiten konncn nicht scbrecken, wen .s das Wohl des Staatcs 
g.It, abcr //,„«-„•„„.. i„ waldwirtscl,aftlicl,e„ Dingen lassen sich nicht 



Waid und Wasserwirtschaft. 

VON 

Dr. Hefele. 

Kgl. bayr. Forslnieislcr u. Professor in Tokio. 

(Voi'trag gc ha I ten in dcr dailsck, Osl- Asia/, GcscllsJiafl fi'ir h'attir v. Volkcrkunde 
in Tokio. '■^ Aits den Mittheiltingen der D. O. A. G. f. K. u. I'^J 



Jedcr Kulturfortschritt ini Lcbcn dcr Volkcr bcruht nicht in letztcr 
Linie auf cincr intensivcren Ausniitzung dcr natiirlichen Giiter, wie sic durch 
klimatischc, oro-und hydrographischc Vcrhiiltnisse des von cinem Volkc 
beswohntcn Landes dem Menschcn von dcr gottlichcn Vorsehung beschicden 
wurdcn. 

Die notwcndige Voraussctzung hicfiir ist zwcifcllos die Erkcnntniss dcs 
Werthes dersclbcn und die nienschlichc Einwirkung ist mituntcr in 
ausgedehntem, ja staunenswerthem Masse moglich, hiiufig findct sic aber 
audi cin schncllcs Ende, je nachdem sic in Ubcreinstimmung odcr in 
Dissonanz mit den " cwigen Gesetzcn " dcr Natur ihrc Wege einschlug. 
Das Grundfundamcnt cines ivahrcn Fortschrittcs muss also die Erforschung 
der Ursaclicn cincr Erschcinung bildcn und so schen wir. dass mit der 
zunchcnicndcn Durchlcuchtung bislier dunkler Gebicte durch tlic alles 
durchdringendcn Strahlcn cincr logisch vorgchenden exactcn Wisscnschaft. 
naturlich audi dcr ^^'eg zum Ziclc geradcr und frcicr von irrcfuhrenden 
Seitenpfadcn wird. 

Gcradc die Entwicklung der Naturwissenschaften in ilirem rapiden 
Ciange hat dcr unanthaltsam vorwiirts hastcnden Zeit bei der Kulturmission 
die unschiitzbarstcn Dienste geleistet, wic cin Blick ins tiigliche Lcben ohne 
weiters Ichrt. 

Dass manch altc hcbgcwohntc X'orurthcile oder Annahmen dabci zu 
V ^\\ komnicn uml nianche trotzicfo Burc cincr Scheinwahrlieit ihrc 



346 Dr. Hefelc: 

Jahrhundcrtc alten Grundfesten untergraben findet, ist nur natiirlich, aber 
ungern trennt sich der Mensch von ciner durch Alter geheiligten 
Uberlieferung. 

Die Besprechung des vorliegenden Thcmas wird zeigen, dass es in 
seinem Gegenstande zii einem der heissurnstrittenen gehort, da eben eine 
allseitig einwandlose Losung der damit vorkniipften Fragen noch weitererer 
exactcr Forschungen bedarf, wenn auch das Endziel in grobcn Umrissen 
schon jetzt klar vor Augen steht. 

Wie aber die Ungleichartigkeit von Kriiften allein eine heilsame 
Storung des starren Gleichgevvichtes 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 
grundhchen Untersuchungen, die Vorbedingung geschaffen werden fiir das 
Aufsteigen des Phonix " Wahrheit und Fortschritt " aus der Asche der im 
Kampfe zerstorten Vorurtheile. 

Der Grund warum bci der eigenthcli beabsichtigten Ausprache iiber 
die " WasscfivirtscJiaft'' und deren Werth fiir ein Land, der " IVald" als 
gleichberechtigt, ja voranstehend genannt wird, liegt in der AbhLlngigkeit 
der Letzteren zum grossen Theile von den Verhiiltnissen des Ersteren. 
Im Wasscr 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 P>denbewohner jene Riesenkrafte leiht, welche 
zur Durchfuhrung der immensen Plane seiner nimmcr rastenden geistigen 
Capacitiit eines der grossten Hilfsmittel darstellt. 

Gewissermassen schon der blosse Anblick der Weltkugel scheint uns 
die Bedeutung des Wasscrs auf unserem Planeten ziemlich eindringlich zum 
Bewusstsein bringen zu wollen, deim 3/4 der Oberfliiche ist allein von den 
Mccrcn eingenommen. Die Bedeutung dieser internationalen I landelsstrassc 
der Volker wlichst mit jedem Tage und die Ausgestaltung der Verkehrs- 
mittel durch die Technik macht sozusagen die Welt nach und nach klciner 
unddic Mission der Kulturstasten grosser und grosser, bringt das Zicl der 
geistigen und sittlicJien Hebnng der Volker der luxle niiher und niiher. 

Um aber in diescm Austausch der realen und geistigen Produkte einen 



Wald niid Wasserwirtschaft. 347 

durch liistoi-ische Vergangenheit namentlich abcr den vcrliehcncn Talcntcn 
Lntsprcchcndcn Platz im Rathe d :r Volker fiir den Verkehr uber die Mecr 
zu behaupten, ist die Enhoicklung der Produktion im Innern der Lander 
eine bindende Voraiissetziini,^ Und hier kommen die Binnengewiisser die 
Seen, die Fliisse iiiid Bae/ie rjur Geltung. Sie bilden iS<i\\ Kernpunkt der 
Giitererzx-ugung sei es direct durch Fructificiruni,'- der im natiirlichen 
Boden vorhandenen Xahrstoffe fiir das Wachsthum von I'eldfriicliten, 
wie im Ac^n-arstaat oder intUrect, als Lieferanten motorisclier Kraft fiir 
die Maschinen des Industriestaates. Stets aber ist die Krzielun- 
einer segensreichen Wirkiing gekniipft an ein gewisses Ebenmaas. eine 
gewisse continuirliclie Bestandigkeit, nirgends sind Extreme nacli dem 
Zuviel wie Zuwenig von solch einschneidenden Folgen begleitet. wie 
hicr. 

Die Austattung eines Landes mit /.aliireichen guten Fliissen und 
Wasserliiufen gibt einen guten Masstab der Beurtlieilung seiner Entwick- 
kmgsfahigkeit gegeniiber einem anderen Lande bei sonst gleichen Beding- 
ungen. Je grosser die Strome, je tiefer dieselben sind, je leichter und je 
weiter sie vom Meere hmdeinwarts befahren werden k^uinen mit grossen 
Schiffen, desto werthvollere Verkelirsmittcl stA\y:\-\ sie dar und. wenn es audi 
eine Zeitlang schien, als ob iiire Bedeutung chn-ch chV- Eisenbahnen in ^\^n 
Hintergrund gedrangt wurde, so hat eben ch'e neueste Zeit mit ihren 
Riesenprojekten. z. B. dem MitteUandkanal in Deutschkand bewiesen. wie 
gerade bei stark entwickeltem X'erkehr nw^i wirtschafth'chem Aufschwung die 
Wasserstrasse der grossen I-'liisse u. Kanale wegen ihrer liohen Leistungs- 
fahigkeit und BiHigkeit, speciell bei im Eigenwerth niedrigen Giitern, nicht 
entbehrt werck-n kann. 

Wo ^[a!!gel an natiirhchen W'asserwegen herrscht. wird sich das 
I'.isenbalmnetz bei" giinstiger Aussicht der Produktion verdichten aber 
immer wird es auch mit ciem Xachtlieile gross^Mvr Kosten behaftet sein 
Die gute Heiiutzlxu-keit ck-r W'asserstrassen hangt jedoch nicht zum 
geringsten Afasse von regelmassigen W'asserstiinden und mogHchst un- 
veniuderlielien Sohlenverhaltnissen ab. \^a es bei ^\,:n grossen Stnunen 
vor allem der Oberhiuf und che Seitenllusse sind. welche die giinstigen 
Oder uug-ini^tigeii X'eriinderungen wranlassen. m\^\ eine l-a'nflussnahme von 



34H 



Dr. HcIVle: 



Scitcn clcs Mcnschcn im Grosscn (^anzcn Jticr cinsctzcn muss, S(i crL^icbt sich 
\<.n\ sclbst dcrcn i^rossc W'icliti^kcit im Systcmc tier W'asscrwirtscliaft. 
J3ic im XaclifolgeiKlcn bctrachtetcn wirtschaftliclicn Vortlicilc dcr klcincrcn 
Fliissc iind Wasserlaufc wcrden von den i^rosscii Verkehrstromen, bis zu 
einem ausi4'!ebiLjen (irade i^etheilt, so d^ss ilire gemeinschattliche .Ynfi-'ihrunsj; 
z\veckmassiL( erscheint. 

W'ahrend bei tlen kleincren Fliissen imd Wasserlaufen der W'erth als 
i^rosscs l^crkchrsinittcl dcs llaiiilcls \veL;fallt, leisten allc WasserUlufe fiir 
Laiuhv'irtschaft uml die fiidustric nach mehrfachen ]vichturii_;en Unersetz- 
liches. 

Die Landwirtschaft kann sie nicht entbehren im Sinne : 

l) tk'i' iMitwasserunL^'. 2) Zum Zwecke der kJewasserinii;" 

& k^'uctification des lx)dens, 
walirend die xom Gewerbe u. der Industrie so selir t^X'suchte billit^-c 
l^etriebskraft \\\\i\ die Mogliclikeit eines billii;"en Lokaltransportes fiir 
Rohprockikte, bei _^'77V.sji'r/77' k^ntwickkm^" des lainh^nrtscliaftlicJicn l^etriebes 
selbstverstandh'ch von demselben tku'ch die Zuhilfenahnie der Masckiinen 
etc. im Sinne ckn' i^ieiclien X'ortheik', wie sie die intkistrielle TliatiL^'keit 
sucht, ebenfalls t^etheilt wird. Die Enti^sascnuii^- dcs Laiida durch die 
Jdiissc ist L^ewissermassen ihre natiirHchste .Vufi^'abe, dcww das meteorisch 
niederstrcnnende Wasser muss diesen .\.us\ve<^ haben, wenn niclit ein 
llinderniss der Kultur und ckimit der Jiewohnbarkeit eines Landes durch 
WM-sumpfun<4- [^^eschaffen wertlen soil. Die . I /dl/fSS£-/-(h'Sc'/i der I'di'isse sind 
von verschiedenen UmstLlnden bedin;4t. in erster Linie von -lev .lfr//^<^-c- dcr 
fallciidcn Xicdcrsihlai^-c in ilirem l'a'nzuL;'s_L;'ebiete. \Vo diese in einem 
Dande nur _i,'777 //.;,'■ sinel, wird man veri;"eblich nach Wasserlaufen \o\\ einit^er 
HedeutuUL;' suchen. 

In Aden z. Ik Hdlt durchschnittlich alk' drei Jahre einmal ausi^iebij^er 
ReL;en uiid die \'orkehrui)_i;"en durch kiinstlichen Ausban natiirlicher I'els- 
beckeii im (iebir^e als W'asserreservoire, um nur das nr)thi^\- Trinkwasser 
Uir die spiirliche ikxolkeruuL;" zu eriialten, sind w eltl)ekamil. Der 

Missisipi der i;e\\aItiL;ste Strom Xordamerikas mit dem unL;\'heuren kanzui;s- 
s^ebietc von 3,150,000 L '. Kui. fiihrt daj^et^x-n xon (V-\\ pro Jahr ca 90,000. 
Millionen enL;l-Cubickiss betraL;endeu Xiederschlat;en auf diesem 



>Viil<l II 11(1 ^Vil^^('^^vil•ls(•Il;l^. "4'^^ 

Tcrritorium, ca 19,500 Millioncii Cul)icfii.ss ilcm Mccrc zii. (2$% clef Xicdur- 
schliigc.) 

Die Ricscnsti'fimc Siulamcrikas (Amazoncn & Laplata-Stom/ mid dcs Ob. 
dcs 54r()ssten asiatisclu n Stromcs in Iviissland, dcs Vantzckiaiii^ in China 
lassen in ilit\n L;"c\\altiL;'en W'asscrmasscn cincn Riickschluss anf die Mcnj^c 
der Xiederschli'i^e in ibreni I^inzui^sj^xbiete sowie aiif dcssen Cirossc zii. 
I'^reilich ist die Wrtlu iluiii;- der Xiederschlai:(smen_L,an in (\Qn ant^^efiilirten 
Sti'onieinzu_L:;s<4-ebietcn selbst eine uniso un^leiclimiissiL^erc, je kliniatisch nnd 
orooraphiscli verscliiedeiiere Re_i4"if)nen umschlossen bzw. drainirt u\:rden. 

Das ]'crhaltniss z\\\^z\\*^\\ Xicdtrschlagsvicni^-c unil Abfiihnnasscn ist 
keineswet^s ein koi/staiitis sondern. wie leiclit erkliirlicli, von i\i.:n 
variablen \'erh;dtnisscn des l^otjens nach natiirliclier l*'orm. piu'sikalisclier 
]?cscliaffenkcit etc aljlii'ini^i^" und am lun'strii bedini;t durch cKas Fclilcn 
oder ]\>rl/irii(/ciisci// einer Pnanz(.'nik'cke und deren Zusaiiniicnsctzuiii^. IV-j 
(K r I'esnreclnnin" di s l'",influsses dv,s W'akks werde ich liierauf ansliihrliclier 
zuruckkomnien. 

W'a^ tiic (iraiiiicriiuh- 'I'ltatii^kcit <Lt l^lussc stort, wird naturnot- 
wendi^er W'eisc auch dii- i^iinsti^c W'irkiiiij^- der DrainaL;"e beeintr;icliti_L,fen. 
So selien \\ir (k-nn iiberalK nanientlich in (\^\\ mittleren \\\\i\ untcrcn 
kdusslilufen niit (\(:\\ L;erinL!;en ('lefldlen. woselbst in ckr Rej^el die l-^b».iic 
ckis Charasteristicuni ikr anstosseiuk-n L;iiulschaft bikk-n wirck tUirch 
/ V/Ti'/A/r/v///;'" der /•'/iissc\ also bei iii(Vii^i-/i/tfi-iii l-.inLjreifen des M^nsclien 
Uberscli\\eninuniL;\n, ka'sstopfuni^en beini l'"is<j^anL;". Xeii^inv^" zur Hv.tt\"erle- 
i^niiL;', X'ersaiukini;' cte. in alien k'idlen aber eine starke Wrmintlerunj^' der 
Abli'ihrun^" tks \\ assers eintreten. k's muss also ilie St^r^e einer Laiulcs- 
verwaltuiiL;" auf eine C(>riwfi<>i! und VhiTZ^nr/iiiiii^- ihrer W'asscrlaufe 
i^erichttt sein. natiuliclie Abtlussbindcrnisse niiissen beseiti-t wwd kiinst- 
liche iMiibauten aut iln\' \\ irkuni; in ttua nachtheilii^em Sinn • 'uxor wi.hl 
<.;ej)riirt werden. 

Die zweite \\ielitii;e Autl^abe des l-dusskiufe niimlich ik r " i\'ii.' iis Si- 
nn/ 1^" des Kulturlands li.it eine weitaus _^'7V'V,V(V\- Heachtun;^ von Scitc tics 
Mensclun s^ilunden imd \\\\\\ dies X'erludtniss \volil auch in /.ukunft bestc- 
lun ; die- M(>oliolikeit ^\(V l-'ruktiTieirunL; uni;eheurer l-"kiclu'n ist in viek-n 
k'alkn niclit^ weiter als eine Ira^e der I'.inleitun^ von i;\nu;^vnd W'asser 



\ 



350 Dr. Hefele: 

um niineralisch tauglichen Boden durch die Zufuhrung- der Pflanze nothii^en, 
bishcr fehlenden, Feuchtigkeit, in ertragsreiche Kultur umzuwandcln. 

Das grosste Project der Neuzeit dieser Art, diirften die Nilsperr- 
diimme bei Assua/i bczw. S/?// im oberen resp. mittleren .Acgypten 
sein, welche in Vcrbindung mit den bereits bestehenden bei Kairo eine 
systematisclie Ausniitzung der Nilfluth im Fri'tjabre in grossartigcr Weisc 
bezwecken. Durch Sperrdamme sollen Stauseen geschaffen werden, deren 
Wasser nach Bedarf zur Abgabe kommt inid die Bodenbenutzung in 
ausgedehntem Masstabe, namentlich fiir holier gelegene Liindereien, welche 
mit den bishcrigen Staumittehi Wasser nicht erhalten konnten, garantirt. 
Den riesigen Kosten von 2 Mill Pfund stehen dauernde P'rhohung des 
National\"erm()gens, der Steuerkraft des Landes unci P.innahmcn aus dem 
zunehmenden \'erkehre in i'lberlegener Grosse gegeniiber. 

Btreits ertragsfahiger Bodcn wird in seinem P^rtrage durch daucDide 
Bewasscrung gesteigert, das wird namentlich dort zur Geltung kommen, wo 
sonstigc gi'tiistigc kliiiintisclw Faktorcn iiocli untcrstfitrj.cud cingrcifni. In 
pAiropa hat P^rankreich '}i:w 31. Theil, Italien ckn zwangigstcn Theil seiner 
Landflilche zur I^ewasserung eingerichtct, widirend in unserem I leimathlande 
Deutschtand diese Zifier nicht erreicht ist. 

Je mehr l^ei wachsender Bevolkerung und nicht beliebig vcrvichr-hczii.'. 
cultivirbarcm Boden gcringerc Bcklen der Kultur unterworfen werden, desto 
mehr wird audi das billige Diingemittel des W'assers in .Anspruch 
genommen werden. 

Im Allgemcinen sintl es audi hier wiederum die kleineren u. mittleren 
W'asserUlufe welche fiir solche Zwecke in Hetracht konnen, (\c\\\\ die grossen 
Miisse bediirfen zu ilirer Nutzbarmaclumg kostpieliger Anlagen. welchen 
nicht imnier tlie entsprechendcn (legenwerthe gegeniiberstehen. Was 
(.ndlich tleii Wcrtli von Wasscrlaufen fiir (ici^'crbc iiiul ]]idustric)i betrifft, 
so ftreben (iewerbe, wie industridler Bctriib im engeren Siiuu- nach P>rsatz 
der niLiischlichen Ari)eitskrart (lurch den Motor, tlessen Antrieb \on einer 
m(')gliclist billigeii, Kichl crhrdtlidun nnd nachhaltigen Kraft(iuelle i. e. tlem 
Wasser erfolgen soil. Audi dicseii Auforck rungen entspredien die geringe- 
ren Wasserliiufe besser im Durehschnitt als grosse Str(')me, da letztere wohl 
immense Kriifte liefL-rn aber audi enornie Anlasjeii zur Realisirunu" derselben 



Waltl iiiKl WassonvirtscIiaCt. 351 

crfordern. (Nutzbarniaclum^' dcs Niagnrafalls in Amcrika.j Die i^cdcutun^ 
(Icr Kraft dcs W'asscrs als Antriebsmittcl nimmt zii, jc mchr dcr Cirossbctricb 
dcccntralisirt wird, was hinsichtlich ciiicr yXiizald liuliistriccn /.. 1^. 
] Jolziiulustriccii sclion dcr l'\all ist. 

J)ic AI(')^liclikcit dcr clcctrischen Kraftubertra^imc( ist zwcifcllos als 
die llin\vc<4"r;'uiiniiiis4 cines Ilindcrnisses fiir die Xutzbarnvicliuiij^' mancher 
Wasscrkraft zu betrachtcn. 

Die \Vasserleitun|L;cn der grossen Stiidte greifeii endlich zuri'ick bis in 
die Oucllengebictc, also deu Anfaiig" allcr Wasserlaufe, iind die Wirkungen 
(k^rselbcn in sanitilrer Jlinsicht bediirfen kcincr niUieren l^egrunduiiL;'. 

Den Seen Koniint jene JiedcutunL;" wie den Flussen & Biichen wcnigcr 
zu, sie sind nielir iokalisirt, es feldt ihnen die Moclukition tier Lilngciicnt- 
wickknig', obwolil sie untcr Umstiinden ini s^leiclien Sinne wie kaufende 
Wasser nutzbar geniacht werden k(")nnen. 

J)iesen S(\q-(-//srcic//cii Wirkungen des W'assers der l^inncnfliisse etc. \on 
denen cine einzige gcniigt, uni ilinen (He ICigenschaft tier Unentbehrlickeit 
zu verleihen, stelien Verheerungen und Katastrophen t^egenuber, welchc in 
manchen Liindern (Uu'ch ihre hiUifi^e \\ ietlerkehr die Ix-wolinbarkeit und 
Cultur weiter 1 .antlesstrecken anindlierten und L;anze (k'L;enilen dc-r 
\'err)dun_<4' preis^aben. 

je nielu' die \'(")lkcr der JCrde dieselbe occupieren. je dicliter sie auf 
gegebeneni Raunie sich concentriren, desto mekr wird man sich der 
Jk'kiinipfuiiL;" des tien Iv^den bedrohendL-n l^ngeheuers widnien. 

W'olil wird es nienials nii\<:;"lich seiii. eiL;entlielie Katastropfen grr>ssten 
Stils, welclie in Sttnun^en der Atniosphiire cte. ihre I'rsaclie habcn <iclcr 
eiiier \ eriinderunt;- von bislieris^en (ileiclu^ewichtsfaktoren iin W'eltsysteme 
zur Last faUen ni<)ocn (^uian denke nur an die I'heorie der allniahliijjen 
AbkiihluiiL;- der I'-nie bis zur \'ei-eisunL;', an i\^\\ k'.intluss iler Sonnciiflcckcii 
etc.) abzuwenden. aber bis /.u eineni L;ewissen (Irade besit/t der ilenkomlo 
Alensch Mitlel. inn sich zu scluitzen und zu wehreii gei^en i\K:u \'ernicl)tuni^s- 
kani]>r, den uni^eziioejti,- Nat urkri'ilte L;e_i;en ihn h'lln-en. u. wenn audi nacb 
nach dem I )ichterw()rt(.- " deini die l-.leinente hassen das (lebilil von 
Menschenhaiitl," alles irdische Streben eitel zu sein sr/iiiiit, so tlurfeii 
wirilochdie 1 liinde nicht in dvu Schooss K-i-en u ml niiisser. das J/<v/j>"< "//<■//- 



352 Dr. Hefele: 

nids^licJic 7.\\ errcichen streben. 

Man hat sich in ncucrer Zcit nicht niclir bcc^nugt mit dcr lickampfun^" 
(Icr l^Vjloccrschcinuni;", dcm Hochivasscr als solclicm. sondcrn ist dcr 
I-'ra^c nacli den UrsacJicii nahcri^cruckt. J licr findct man nun in dcr 
Litteratur den \'oni Volksglaubcn lilngst als Ursachc bczcichneten 
J laiiptfaktor, ncmlich den Zustaml des Ur.sprun<(st(cbietes der Fliissc 
In'nsiclitlieh seiner Pflauzciidcckw und in der llauptsache ist der " W'ald " 
(kaniit i;emeint, in 1 lerz und Xieren £4"epruft auf O^^w Zusammenlianq; mit 
den vcrhcercnden W'irkun^en der W'asser in (\vw l^diissen. 

W'ir sind (kmiit unwillkiirlich in jenes (lebiet der W'asserklufe eing'ctrcten, 
das uns k'orstleute naturs^cmilss am meistcn inte-ressirt, in C\.i^\\ Obcrlauf, tlas 
aber audi, wie ich thirzutlum lioffe, die meiste Aussiclit auf crfolgrcichc 
jH-('iiiJIi/ssinio- dcr heute so brennenden "Wasscrfrat^'c" _q;ibt. Icli beschrilnke 
micli tkilier im i^rossen Gan/.en auf die J?esprccliun_L;' der m(")_<4lichen 
k',in\\'irkun<;"en auf die \VasserUlufe, im Jiiisfc-/iiiii_ii-s-(jcbi('ti\ welches wohl 
meisteiis im Ciel)ir_<4e zu suchen ist und auf (h"e lick;lm])funi;' der VJhc\ in den 
oheroi im ("iel)ir_L;e c^elcgenen Thcile der hdiisse. wo die i^ro&soi GefaUc 
up.d inslKSf)ndere (h'c CiCSclticbcfi'iJiruni;' die meistcn Scliiidcn vcrursaclicn, 
widu'cml im mittleren und unteren l.aufe der Strome iiirlir che JAv/^'V der 
/u_L;"cfii1n'tcn W'assermassen verderblich wird. 

Dcr crsterc Tlieil licsitzt \icleicht nicht immer jene riiumh'clu- Ausdcli- 
inm<4" der Scliiidcn, wie sic (h'c \'i(.;dcrun<4en der .AriiniUmL;si;"ebicte u.nd die 
Mittellilufe der I'di'issc aufweiscn, aber an lutciisitiit der \'er\\iistun_L;- kommt 
kein andcrer Iheil ihni L;leich. 1 licr ist (kis l\'rrain (Kr W'ildwasscr, der 
" W'iliiluulic^' und ihrer \'( rmulnnmi^en und jeder ImToIl;" der hier errun_L;'en 
wird, kommt i\i:\\ .\ie(krun;;cn der I'^bene /u (lUte y\\\(\ /war in potenzirtem 
Maase. 

I''rilj4t man ziini'ichst, che man (V-w \\\rt dcr " Ju'waltUuiL;" "' untcrsuolit, 
ob und in iK'ii-iCcit cine rnanzen(k'cls(,- im I'rsprungsgc-bictc uberliau]H cinen 
k.induss ansiibt auf ikis Ktt^ime (Ur W'asser, so konmit man auf (irund tier 
rnt(.-rsucliunL;(H (ks ba^'rischcii I'rof I'lheriuaycr, dts lei(k-r zu friih 
v'crstorbeni II I'.. Wolhi)' in Miinclien u. Anckrcr zu (k'm Schhisse, (kiss 
/>/■/<• rnanz<n(k c Isc (iras, //urn] ciiuit \'(>i:::iij^ L^et^cniibcr /v^/zAv// (icbir_i;c 
Iti'dciitct. 



WaM iiiKl Wassoiwirtscliafl. 353 

Unsclnwr ist cinzuselKii, dass jcdcr l-iodcnLibcr/.uj^^ pcrcnnirciidcr 
(k-wiiclisc eiiKii L^'cwisscii Aus;_,deicli in dcr Wusscrfuhriin'^ licrvorruft. iiidcni 
die ()ber-\vic uiitcrirdischc Wasscrablcitunj^ vcrzo<,rci't wiril, was in tier 
( icsainiiitw ii'kuiii^r ^n^lciLliinassigcrcr WasscrstLiiidc seincn sclilicsslichcn 
Ausdnicl< fiiuLt. 

Im'ik: \'crlaiiL;sanuin_<4" dcr obcrirtlisclicii Abfulir bcdcutct clnii cin 
bcsscrcs liiiisickcrcn dcr nictcorischcii Wasscr in den cku'cli die I'flanzen- 
wurzeln pli)'sikaliscli L^i'instit^er d. li. lockerer i^emacliten Hoden, \V(jdin"cli 
n/i/-(/t/(lssi\>^'kcit II /id Aiifnahinsfa'tigkcit sich stcii^ern. 

I'"ur die wieditige iM'a^e dcr Qitcllciibildiin<^- ii. ILrlialtniii:; an ^\k:\\ 
I lani;en dcr (iebir^e komnit in erster Linic das lant^ere \"er\veilen des 
oberirdischen W'assers auf dcr betr. I'Ulclie in l^etraclit, dennyr mclir da\"on 
in (\k:\\ l^odeii einsickert, desto bcsscr wird die Spcisuiii:; dcr OitclLn 
crfolj^-cii, die ja niclits weiter sind, aks tiefer /u 'I'a.^e ti'etendes Sickerwasser 
b()lierer Lai^en. Sicht man \'o\\ " W'ald'' xorerst ab, S(i ist ini Ubrij^en bei 
(\c\- I[(">/un-L(riic dcY JLiiirjiii^sg-fbic'tc nieist nur eine Jhisc/iz'L\!^\ia(ii>/i mid in 
dcr I faiiplsachc "(iras" als l-5odendecke vorhandcn. 

(iras erfiiUt aber ^\k:\\ Zweck der W'asserx-erlant^sanniwii^- u. der X'erluit- 
uni;- ties 'rerrainano-riffes /////- scJir niaiiii-clhaft , deiin cin diclitcr (irasshi/. ist 
nielit L^enu^end durcldiissit;' und lirini^t nanientlicli in steilen Lii^en cin 
r(ipi({cs obcrjiiiclilichcs Abjlicsscii niit sicli, ist also zweifeilos fiir (ileicli- 
niLissi^kcit iler Wasserfulirnni;" der Fliisse uikI fiir (Juellen \on iii/cn'orciii 
W'ertlie, wenn aucli besser als iiaktcr Hoden. Lockcrc (jiuisdcckcu {//(inH 
sind aber keineswei^s i;et;en tlen An^^rilV iles W'assers L;esichert. wie der 
i\u_<4enscliein in japan lehrt. 

Als man in I'rankreich 1804 iiolTte, eine Jhrit/iiiii/iii^- der j^elVihrdetcn 
I'kulien im Heri;lande diircli lUiRASUXCi dcr voti W'ald ciitldosstcit, 
(/iirc/i die meteorischen Wasscr in Kewc^uni; i;eratlienen, und die Wililbachc 
mil ScliuU Uillcnden Heri;luinL;e zu erzielen. ila erwieseii sieh die d.iiMn 
L^ekniipftcn 1 lolfnunj^cn als truL;eriseh uml es blieb niclits amleres ubri«»" 
als zur .}i(//'(>rs/inii^- zu j^reilcn. leh tlenkc. ilas ist ein deutlielier l-'ini;er/.ci«'' 
Uir ilie Zukunft der wir/.. H. in lapan ent<;\\i;enst uern.wenn die riicksichtlose 
\'erselilecliterun;4 der //(//</ in steilen Lai^en '.veiler i^efiilirt winl, abgcsohcn 
da\on, dass, wie crwiilint, der (iiiisiciic/is an si'c/i, auch woiiii er den 



354 Dr. Hefele: 

l-iodcn ini concrctcn Fallc lullt, dcnnoch fiir cine i^'vvV.v.vr/r Regelmassigkeit 
cicr Wassci'stilndc iin Sinnc dcr A'crlangsanuin;,^" dcs Abflusses des 
.\tmospharilicu imd soniit dcr rjcillichoi Vcrtheilitiii:; dcr Wassermassen 
starker Regen cte. nicJits llervorrgcndcs leistet. 

In Japan muss aber der Fragc rjivischcn dci/i Zusaminciiliajigc dcr 
Bcdcckitiiii- dci- (Ic'birgc mit ciilsprccJtciidcr Vegetation unel den Jldssc/-- 
vcrhalUiisscii eine erhohte Aufnietksanikeit geschenkt werden aus .czlvv' 
(iriinden. 

i) Infolge <\v\- schuialcn Ajisforniniig des Landes mit seinen der 

l.iingsrielitung fo'.genden ziemlieh hohen (jebirgsziigen ist die 

Liingenentwieklung der meistcn Fliisse eine sehr gcrlngc, es 

lierrseht ilas bei grosser Lilnge von Fliissen in anderen Liindern 

auf tk-n oberen Lauf besehrankte ^tcilc Gefalle untl der " Wildbach- 

artigc Charaktcr'' I'or, ja verklsst sie zumeist nicht bis zn ihrer 

k-inmutukmg ins Mcer, wie man an (\c\\ Verb.altnissen einer .\nzakl 

derselben J'^ijigawa, ( )iga\va, Kisogawa ete. cte. beobachten 

kann. 

2) Die Ansfoiiintiig der ] lange im (lebirge ist in diesem \olkanisehen 

Fande eine extra s/ci!r ; tlie erodierende Thatigkeit des auf diese 

Alt raSihcr wie sonstwo zum .\bflusse konimenden Niederschkigs- 

wassers wird im uuguiisligcn Sinne nocli untersti'itzt durcli die 

durehgangig sehr weiehe V'erwitterungsdeeke alter Schiefer etc, 

welche dem (irundgesteine aufliegt. 

\\'(.nn ujiti^'r (Kr bishcrigen l^enutzung als Hara ein grosser Theile des 

Hergkuuks in Japan sclieinbar nock nicJit so sichtbar gelitten bat, als man 

nach dem ("lesagten vermuthen konnte, so ist zu bcdenkcn, dass wir fiir die 

dem Durchschnittsreisenden zu Augen kommenden nietlrigeren Vc^rberge 

einr dureli das Klima bcdiiigtc ausserordentliche Rcgcncratioiiskraft der 

V^egetation habiii. 1 )iese Kommt aber fur liolie, kiihlere Lagen nicht mehr 

so sehr in IVtraeht. I)eutlieh erkennbar siiul auch schon fiir (\c\\ Laien die 

umfangreichen Terrainzerstrjrungen im Herghuul des mittleren \\\\(\ si'idlichen 

Ja])an. \'on Kobe bis Shimonoscki prilsv. iitiren sieh die Ik-rge im liuidigoi 

Zustandc- (1. h. es ])linkt der rothgelbe. nakte (irund der lliinge durch tlie 

zerstorte Ifaia und (\ki\\ W'ald. Wer aber erst einen Blick in tlas Iiiiicrc 



Wald iind Wasserwirtschaft. 355 

gethan hat, dcm ist kein Zvvcifcl iibcr die unhcimlichc Thiitigkeit des 
Rachegcistcs dcs untcrgegangcncn Waldes. 

Es blcibt nun zu untersuchcn, in wic wcit dcr " JVa/d" als dcr Ausdruck 
der hcichstcn organischcn (jcstaltung dcr Bodendcckc der Erde cincn 
Einjiuss auf die Wasscrvcrhiiltnisse cincs Landes hat. Diese Frage liisst 
sich nicht bcantwortcn, ohne dass man die Sachc in zwei Abschnitte thcilt, 
namhch in die sog 

I. ^' IValdkliniafrage" d. h. den Zusammenhang des Waldes mit 
dem KHma eines Landes ini Allgemcinen oder auf concreten 
Orthchkeiten und, 

II. Die IVirkitng- des Waldes, u. zwar des Bergii'aldes in erster Linie, 

auf Regelung des Wasserabjlusses u. Geschiebefiihrung. 

Dcr Glaube an cine hcilsanie Wirkung des Waldes hinsichtlich eincr 
Verbesserung des Klimas durch Milderung der Tcniperaturextreme, \'er- 
mchrung von Niederschliigen, Verminderung der Hagelgefahr etc., kurz als 
klimatischen Factor ist cin sehr alter, nichtsdestoweniger jedoch jetzt als 
eines der unter deni Ansturm der cxacten wisscnschaftlichen Forschung 
gcfallcnen V'orurtheile zu betrachten. 

]\Ian hat auf die Trockenheit resp. Rcgcnarnuith der Mittelmeerlander 
ini Zusamnienhangc mit ihrem geringen Waldroichthum hingewiesen, das 
Beispicl von Italicn & Griechenland angcfi'ihrt. wo grosse Lilndcrstreckcn 
durch die mangelnden Niederschliige zu '_Wusten geworden seien, als 
man den schutzenden Wald zerstort hatte und kein Geringerer als ein 
Alexander von Humboldt hat auf die .\bnahme dcr Regenmcngc und dcr 
Luftfeuchtigkcit durch die Zcrstorungcn dcr Waldungen aufmerksam 
gemacht. Die reichc Zahl dcr Anhanger dicscr Anschauungcn wics die 
hcrvorragendsten Namcn dcr Wisscnschaft auf, aber als man dcr Sache 
durch die metcorologischcn Stationcn imd (.Icrcn Bcobachtungsresultate 
nahcrtrat, nanuiUlich aber durch die Frgcbnissc dcr I'ntcrsuchungen iibcr die 
k>xistenzbcdingungcn dcs Waldes, da gclangte zu dem anfangs Stauncn cr- 
rcgcndcn Rcsultatc, dass nicht dcr Wakl cincn lantluss auf dass Klima habc, 
sondcrn, dass ilie Baum- und Wakivegctation \ollstandig von den 
klimatischen Wrhaltnisscn eincr C^icgcnd abhangig sci, so dass die bishcr 
iiblichcn obencrwahntcn Annahmen nur thcilwcisc richtig sind. 



356 Dr. Hefele: 

Der Wald ist namlich weder naturlich vorhanden noch jemals kunstlich 
bei sonst giinstigsten Verhiiltnissen dauernd begriindbar, wenn z. B. wiihrend 
der Hauptvegetationszeit (in der nordl. gemassigten Zone ]\Iai — August) niclit 
in minimo 50 millimeter Niederschlag auf die betreffende Gegend fallen. In 
Aden 3 B. wird niemals eine Baumvegetation ohne kiinstliche Bewiisserung 
denkbar sein ; die Ebene Californiens verdankt ihren Ruf als Obstkammer 
Amerikas nur den kiinstlichen Bewiisserungsanlagen ; die ersten Ansiedler 
trafen kcinc Biiume an, da eben das nothige Minirintni an Wasscr dem 
Boden bei sonst eminent klimatischen Vorziigen nicht geboten wurde. 

Baumwuchs ist aber trotz geniigender Bodenfeuchtigkeit(:Niederschlage;^ 
cbcnsozvcnig denkbar, tccnn die notwendige relative Lnftfeuchtigkeit von 
mindestens 50% zcdhrend der Vegetations.'^eit nicht vorhanden ist. 

Da wir auf diese zweite Bedingung kaum je Einfluss gewinnen, so ist 
das Bestreben der Griindung von Wald absolut aussichtslos in diesem 
Falle, mag die Bodenfeuchtigkeit noch so reichlich vorhanden sein. 

Beweise fur diese Thesen finden sich durch die ganze Welt in 
priignantem Ausdrucke. 

Die Graspriirien im Centrum Nordamerikas und die Pampas Siidamerikas 
und Australicns, die Wilsten Afrikas, Asiens, (Gobi in China) sie konnen 
nie und nimmcr durch des Menschen Hand auch mit Aufwand aller Mittel, 
Baum oder Waldvegetation tragen, da die Luftfeuchtigkeit in geniigender 
Menge nicht gcgcben werdcn kann. 

Das Sinkcn der relat. Luftfeuchtigkeit unter ^0% wahrend der Vegeta- 
tionszeit kann nur von Gras odcr Staudengewachsen crtragen warden, (Priirien 
Amerikas n. Australians) wahrend cin Herabgehen der Luftfeuchtigkeit unter 
^0% rel. P'^euchtigkeit und eine Niederschlagsmenge unter 20 ?/////<? wahrend 
der Ilaupt-Vegetationsmonate die vegetationsloscn Wiisten (Asiens u. Afri- 
kas) zur P'olge hat. 

Xur auf <\cn Grenzgebietcn von Priirie und Wald ist anscheinend cine 
geringe Verschiebung in Sinne der iVusdehnung des W'aldes vioglie/i, aber 
bei genaucrer Untersuchung zeigt sich immer, dass man ehemaligcn 
Waldgrund vor sich hat, der durch irgcnd eine gewaltsame PLinwirkung 
zu Grunde ging, was beispiclsweise in Amerika im ausgiebigsten Massstabc 
bemerkt werden kann, indem es durch die llilfc des P'eucrs Icidcr 



Wald iind TVasserwirtschaft. 357 

i^elang-, den Wald auf ciner dirckt nord-siidlich streichenden Linie, parallell 
dem Missisippilaufc zuruckzudriingcn u. zwar um x'olle lo Liingengrade 
(100^-90° wcstl. L.), cine gewiss nicht zu unterschLitzende Leistung ! 

Dass man hicr.zitlandc in z. B. Hokkaido durch das schonungslosc und 
Linvcrniinftigc Anziindcn der Rodcniiberzuge dcs IValdes in der Xilhe der dcr 
Ackcrkultur qcwoniicncn Thillcr das amcrikanisclic l^cispicl, wcnn auch im 
schwachcrem Masstabe, aber mit eincr fiir den .\''^7rt'(!7:c'^/(y ebcnso totlichcn 
W'irkung wicdcrholt, babe icb schon in cinem friihcren \'ortragc crwidint. 

Wedcr die hohcn Tcnipcraturgradc im positivcn Sinnc, wclche in den 
sonnigen Wiistcn .\frikas hcrrschen, schliessen bei gcgebenen Beding- 
ungen dcr FciicJitigkcit dew Wald aus (Oasen), noch Wintertcmpcraturcn 
von — 30 u. 40^ C, wie man sic in America besbachtet hat. Ein grosser Theil 
Sibiricns mit seincn Wintertempcraturen von oft- 45° C. ist voll der reichsten 
Waldschatzc. Erforderniss ist einzig und allein cine mittl. Tcmperatur von 
+ 12 bis i^ C. wiihrcnd dcr Hauptvcgctationszcit. W'ird diesc nicht 
crreicht ( + cS' bis 12^ C), so sinkt dcr Wald zur Staudcn u. Buschform hcrab. 

Tnttjcdcn Monat dcs Jahres Frost cin, so ist J'cgctatiofislosigkcit die 
Folgc. InucrJialb diescr natiirlichcn Existenzfaktorcn gHedert sich der 
Wald nach Form und Zusammoisctzung aus Artcu, den lokalot klimatisc/icn 
VcrJialtnisscn cntsprccJicnd, und da d\c jciccils nordlicJicr bezw. siidlicher 
\om Aequator gelcgcne Zone ;//// cincr niiJicr gclcgcncu. abo- iu grosscrcr 
I'Jcvation bcfindlichcn corrcspoudirt, so ist Icicht zu crkcnncn, wie nach 
rclativ cinfachcn Gcsctzcn die Bcdcckung dcr Contiiicntc durch Waldjlora 
zu dcnkcn ist. 

Wcnn dcm Waldc auch ein gcncrcllcr EinJJuss auf das Kluna iibcr 
ganzc Flussgcbictc und Landstriche odcr Continentc abgcsprochcn iccrdcn 
muss, so darf doch die Miiglichkcit eincr Einn'irkufig auf klimatische 
X'crhaltnissc im Siiinc /oca/cr Modulation cbenfalls nicht iibersehcn werden. 
VAi\ sogenanntcs '^ f,okalklima" kanii. ilhnlich wie an cinem grosseren Sec 
besonderc Luftstromungen hcrrschen. vom Waldc bcinjlusst ivcrdcu und 
wird sich natiirlich ilics umso mchr gcltciul machen. um jc grosscre 
Waldflachcn cs sich hanticlt. 

Die Klarlcgung allcr diescr \'erh;dtnissc muss den eingclciteten 
\ ersuchen iibcrhisscn blcibcn und wcnn dcr Kampf licr Meinungen dariibcr 



\ 



35^ Dr. Hefele: 

audi in den berufenen Kreisen noch lieftig tobt, an dem entscheidenden 
Hauptmomente, dass der Wald klimatische Katastrophen nicJit verhindern 
kann und keincn Einfluss auf VernieJirung u. Vertheilung der NiederscJiliige 
etc. Jiat, ist mit alien Consequengen kcin Zweifel. 

Der Hanptzcerth der W'aldbestsockung' fiir^r/T^^r/Zr/r WasserstLlnde der 
fliessenden Gewiisser und damit fiir die Verhinderung der zerstorcnden und 
verdcrblichen Hochwasser oder fiir Erhohung der fiir Kultur und Industrie 
gleicJi inisslichen, zu niedrigen Wasserstiinde, Hegt auf einem andcrcn 
Gebiete. 

Beivaldinig erJioJit niimlich unter Bedingungen einerseits die Sickcr- 
ivassennengc, was von hervorragendem Einjliiss auf die Spcisiuig der 
Qiicllen ist, und ste/lt andcrseits von alien Bodendecken das bedcntciidste 
viccJiaiiiscJic Hindcniiss gcgen die AbschiueviiiiuJig der Bodens u. Abrut- 
schung der ScJicedecke bei Lawin.enbildung etc dar. 

Die erste Behauptung der Erhohung der Sickerwassermenge, also der 
Ouellenvermehrung, kann in ihrem I'ollcn Umfange nach dem heutigen 
Stande der Wissenschaft niclit mehr aufrechterhalten werden und damit 
fdllt li'iedenmi cine seit alten Zeiten gehegte Ansicht. Wie weit sie noch 
Geltung hat, werden wir sehen. Die Untersuchungen von Ebermaycr in 
Miinchen und Ototzkij's in Petersburg haben den Nachweis geliefert, dass 
der Untergrund unter Wald in der Ebene unter allem Umsti'inden iirvier 
an Eeuchtigkeit ist, als auf freiem Lande, dass also dem Walde der Ebene 
ein Einfluss auf Vennehning der Bodenwasser im Sinne etwa der Erhohung 
des Grundwasserspicgels u. der besscren Speisung von Quelle nicht zukommt. 
Das scheint im ersten Augenblicke stark zu contrastiren mit Ergcbnissen 
von Vcrsuchen, welchc za-hlreichc AutoritLiten hinsichlcich der Wasser- 
Absorption wniS. Retention des W'aldes und namentlich seiner Streu 
angestellt haben. 

Durcli die IJberdeckung des Bodens mit Wald wird in erster Linie 
dessen oberfliichliche V'erkrustung, teilweise erzeugt durch mechanische 
Wirkung (Eestschlagen durch Regentropfen) verhindert, ferner, als cine 
Folgc der durch die 1 lumuserzeugung der Strcndccke bewirkten besseren 
Krummehuig und Lockeruug der Struktur des Bodens bei sonst gleichcr 
niineralischer Beschaffenheit die Aufnahinsfiihigkeit fiir meteorische '^ieder- 



Wald nnd Wasserwirtschaft. 359 

scJillige erhoht. Die herrschenden niedercji Tempcraturcn im Waldc wiihrcnd 
der hauptsiichlich wichtigen wLirmeren Jahreszeit, — im Winter ist der Boden 
an sich durch Frost undurchUissig oder die Bedeckung mit Schnec als cine 
momentan latcntc Wasserquelle zu betrachtcn, — und die dazu kommcnde 
aus der Transpiration der Pflanzen resultierende grossere Feuchtigkeit der 
Luft (3-10)^^ im Mittel melir c;ec^enuber dem Freilande) wirken im Sinne ciner 
Vermehrun^- der Wassermenge des Bodens indirekt, indem dadurch die \'er- 
dunstung des atmospharisch zu Boden gelangtcn Wassers verhindert wird. 

Fine gute, in richtigem Zersetzungsgrade befindlichc Strcudcckc, welchc 
im wohlgcpflegten Walde niemals fehlt, vcrmindert die Verdunstung des 
der Bodenfeuchtigkcit nochmals ganz bedeutend, so dass die Verdunstung 
im Walde etwa nur 20^'q von jener einer Freilandsfliiche bctriigt. 

Diesen giinstigcn Faktoren gegeniiber, welche die Sickcrn'asscnucngc 
exjierimentell nachgewiesenermassen urn niclit weniger als 2\^^ fi'ir den 
strcubcdccktcn Waldbodcn gegeniiber dem icald/oscn crJiolicn, steht nun der 
aufstockende llolzbestand bis zu einem gewisscn C3ra.dcfc'ind/ich gegeniiber. 
In erster Linie werden von dem atmospharischen Xiederschlagswasser ca. 
25% desselben durch die Baumkroncn absorbiert. welche iiberhaupt 7iicht an 
den Boden gclaiin-cu konncn und ist diese Ziffer natiirlich nach Waldzustand 
Holzart, Alter etc. einer gewissen \'eranderung unterliegend. 

In dichtcn l'"ichtenbcstiinden werden die Absorptionsprocente der 
Kronen bis zu 40 und 45)',^ ansteigen und in lichten Lanbwaldbestiinden 
luigekchrt niedrige Betriige aufueisen. Das bisher nicht gcniigend in 
Reclinung gezogene Abflusswasser an i\c\\ Zweigon und Schaften. welches 
bci Hinger dauerndem Regen nachtriiglich diese Procente um cinige 
lunheiten verbessert, ist wegen der Schwierigkeit der Untersuchung fiir 
verschiedene X'erhaltnisse noch niclit hinliinglich einwandfrei bestimmt. 

Die Vervielfiiltigung der Oberfliiche durch die Blatter namentlich aber 
die unzahligen Natleln der l^iiume ist iler raschen \'erdunstung des 
aufgefangenen Wassers und tlanut tlem tlehniti\en \'erluste tlesselben flir den 
Boden ungonuin giinstig. \'on dcui — nach Abzug von etwa -'J- 30^^ des 
Niederschlagswassers. welches an ^^\\ Kronen der Waldbaume hiingcn blicb, 
uiul weiteren 8-/0%, welche durche Wrdunstung aus dcni Boden in die 
Lult \erlorcn gehen. — iibrigen Ouantum \on etwa rund (yoji^ werden ca 



36o Hr. Hefele : 

25^43" durch den Bodcn entgiltig mtfgcsogcn, u. der Rest zur oberidischen 
Abfuhr gebracht, Jeder pflanzenbedekte Boden nun, ganz besonders aber 
dcr durch die Vegetation des Waldcs in Anspruch genommene, 
erfiihrt eine kolossale Vcrringcning seiner Feuchtigkeit, denn die Bi'iume 
sind die grosstcn Wasscrconsumcntcn und darauf ist eine Thatsache 
zuruckzufiihren. welche gecignet ist, durch falschliche Auslegung eine 
geivissc CiiniJie in wcildkonscrvativ Jiandcliidcn Krciscn herv^orzurufen, ja 
vieleicht in der Hand gewissenloser Volksagitatoren und habgieriger 
WaldschUlchter zu cinem allerdings nicht einwandfreien Sturmmittel gegen 
die ErJialtung dcr Wiildcr benutzt zu werden. 

Prof. Ototzkij in Petersburg fand, wie bemerkt, die iibrigens aus 
kleineren Versuchen kingst vermuthete Thatsache, dass der Gnind- 
ivasscrspicgcl in dcr Ebcnc unter Waldungcn eine bcdcutcndc Seukung 
crfahrc und somit praktisch das Gcgcnthcil von der behaupteten 
Bodenfeuchtigkeitsbewahrung durch den Wald dartliuc. SicheiHch kann 
nicht erwartet werden, dass diese oft sehr bedeutende Differenz des 
Wasserstandes in tieferen Schichtcn zwischen Freiland und Wald zu 
Gunstcn einer rcicJilicJicii Speisung der Sickerwasser und damit der Ouellen 
durcli den ^\'al(l spricht. Pei gleicher geo-physikahscher Beschaffenheit 
leistet also die baumlose Ebcnc mehr fiir eine continuirliche Unterstiitzung 
des Grundwasserstandes und der Ouellen als der Wald. Man ist fiir 
den ersten Lugenblick erstaunt i'lber die drainircndc Wirkung der 
Waldvegetation, aber cinig'c Zahlen mogen beweisen, wie kolossal 
der Wasscrverbrnuch durch die vcgctativcn Proccssc dcr Biiunic sich 
stcllt. 

An sich ist die Produktion der organischen Substanz pro Jahr bei dew 
l^iiumen schon grosser als bei alien fibrigcn Kulturgewilchsen und daraus 
crklilrt sich audi der entsprechend ]v')hereWasserverbrauch durch Transpira- 
tion zur ICrfiillung dieser gcsteigcrten Arbeitsleitung. Die im l^aunikorper 
u. l^Iiittern sclbst aiifgcspcichcrtc Wassermcngc ist ebenfalls seln- bedeutend, 
sie betriigt nach ICberniayer bei einer kriiftig entwickelten 85 jg- Fichte 
(ini Ilolzkcirper \\\\(\ in (X'^n Xadelen) ca rooo Liter; eine gieichalterige 
Tanne hatte i-200 Liter W'asser. 

Zur i'roduktion der organischen Substanz verbrauchte eine grosse 



Wald imd Wasserwirtscliaft. 361 

Birkc in 6 Monatcn nicht wcnii^cr als 7080 k(^. odcr pro. Taj:^. 38 Liter im 
Vcrdunstungswcge uiifl cine 115 y^. Rothbuchc beanspruchte ca 50 Liter 
in g;Icicher Zeit, wiihrcnd bei junfjerem Alter (50--60 Jahre) eine Rothbuche 
pro. Tag. 10 Liter ben'ith.igte. 1^2in Jhichcnhoclm'ald producirt auf gutcvi 
Standorte jiihrlich durclischnittlich 7057 kg. Trockensubstanz, was ciner 
jahrlichen Wasserconsumption von etwa 2,187,670 kg. oder = 2i8 ;//;// 
Wasserhohe gleichkommt. 

Diese Zahlen eroffnen Einblick in den Haushalt der Xatur von geradazu 
verbluffender relativer Einfachheit und lassen die cntsprechenden Schliissc 
auf den inncrcn Zusammenhang der ausserlichcn P>scheinungen zu- 
Nivinicnncltr wird es gelingcn einen schonen alten Buchenbestand zu 
erziehen, wo ihni das Minimum seiner zum Waclisthum nothigen Wassermasse 
nicht zu gutc kommcn kann. Ks basirt somit die ganzc Abstufiing der Bonitiit 
bei gleicher physikalisch-chcmischer l^odcnbcschaffcnhcit fiir cine Species 
zu cinciii gJttcii Thcilc auf den G rii iidicas scr-hczw . Xicdcrsc/ilagsvcrhiilt- 
iiisscn. W'as nun fiir die Ebcuc vom k.influss der W'aldes auf die 
Wasserverhiiltnisse gilt, ist kciiicsivcgs zutreffend mit Zunahmc der 
Erhcbung des Bodens, also im Bcrglandc und Gcbirgc ! 

Mag in den Ebenen immerhin der W'ald vcrriiigcrt iccrdcn auf 
Cnind dicscr l^eobachtungen. besondere Xachtheile grosscn Stils werden 
daraus niclit erwaclisen, lK')chstens class einige Ouellen nictleren L'rsprungcs 
versiegen. Grosserc Ouellen \erdanken ihrc Entstchung und ihre Speisung 
mcist dem Dnickzcasscr der hldwrcii Lagcn untl ^\•erden datlurch, dass 
tiefcrc Pliitze cntivaldct werden, kaum in ihrer Starke und in dem 
hcrvorragenden Werthe, der ihnen fiir Wasserversorgung der Flftssc, 
Wicsenbewilsserung etc zukonunt, beintriichtigt werden. 

Mit jcdciii meter luWierer l^odcncrhcbung aber gcstalten sich die 
Wrhidtnissheinsiclitlich der Hodenfeuchtigkei. im \\ iildc g/ittstigtr, xcHi'/ist 
die ]u-deiitiiiig, iler ]\'ert/t des W'nldes. Nicht nur die Xiedersc/iliigc tnchrci: 
sich, sondern auch ilurch tlie Abualnne der Temf'eratur unvl daniit der 
Verdunstung, sowie ihu'ch die grossere Loekerheit des Waldbestandes 
die Siiniiiie tier der dem /nu/eii verbleibeiiden Feue/itigkeit im W'lic/iscn bc- 
griffen ist. Die Wgetationstlauer in don Hoch!agen ist kiirzer und damit die 
Aiispri'iche der Baumevegetation an das Bodetncasser geri//ger. die sitikeuiic 



362 Dr. Ht'fele: 

Durchschnittstemperatur unci Zunahme dcr relativen FeucJitigkcit bewirken 
audi eine geringcrc Transpirationsgrosse. 

In je hoJierc Lagen man daher in den Bergen ansteigt, desto ivenigcr 
wird die in der Ebene an sich bcdeutcndc Diffcrcn:^ im ^oi^^w-Wasscrgehalt 
cincr bczvaldctcii und nnbcivaldctcn FlLiche, desto iiieJir cntkraften sicJi die 
Vorzvurfe gegeii der Wald als einen Feuehtigkeitsver:;eJircr ohnc Gleichen, 
desto Diehr bekomvit der Wald das Reeht der Existenz und fiberall, wo 
man gegen dieses Reeht sfindigte, waren die Folgen scJirecklicJie. Die 
konstante Speisung der Biiche & Fliisse ohne exeessivc scJiiidlicJie Extreme 
wich stets naeh der Entwaldung den abnormsten Gegensiitzen und die 
Geschichte der Schweiz, Tirols, des sudlichen Frankreichs (Provence) 
Italiens, Griechenlands (und teilweise auch Japans) lehrt jedem Un.befan- 
genen die traurigen Folgen, welchc die Vernichtung des dem JMenschen 
von der Natur gegebenen schiitzenden Waldes nach sich zieht. 

Im Bergland und Gebirge besitzt der Wald eben nicht bios das Reeht 
der Existenz, sondern er wird zur zivingenden Notivendigkeit , da sich 
der Faktor des nieeJianischen Hindernisses gegen die oberfliiehlieh zuui 
Abflitss koninienden Wasser Jiinzngesellt. Die Menge des in ihreui 
OberJidcJienablaufe zerstorend wirkcnden Niederschlagswassers ist ex- 
pcrimentell auf ca Jjj^^ der Totalniederschlagsgrosse erinittelt und einer 
Vergrosserung oder ]\'rniinder2tng in ziveifaeJier W^eise jinterioorfen. Jc 
absorptionsfiiJiiger der Boden ist, auf dem der OberflLlchenabfluss erfolgt 
desto, mehr wird sie verringert und die Anfsaiignngnienge erhoht, von 
der hinwiederuni das fur die Ouellenspeisung so wichtige Sickerivasser 
abhcingt, welches im Durchschnitt 17-20%" des Niederschlages beziffert. 

In positivcm Sinne wirkt nun auf das oberflachlich zum Ablaut kommende 
Wasscrquantum und zwar in ga)iz bedeutcndem Masse die Neigung des in 
Fragc kommenden Terrains. Je ivenigcr lang infolge der Schwerkraft das 
Wasser auf dem concreten Bodcn verweilt, desto weniger versickert, desto 
mehr nimmt die Oberjlachenablaiifzvasserniengc zu. 

Wo Wald fehlt, ist im geneigtcn Terrain, also im Bergland, ein Ansteigen 
der oberflachlich abfliessenden Wasserquanta auf ^,--55 % der Niederschliige, 
je nach dem Tcrrainwinkel, zu erwartcn uud in vegctationslosen Gebieten 
der Gebirge gehoren 60% und mehr keineswegs zu den Seltenhciten. Die 



Wald und Wasserwirtscliaft. 



363 



I 



zerstorende Kraft bcwegten Wassers wiichst natiirlich init seiner Mengc 
und allcs was dieser Thatif^kcit cincn Ilcmmschuh aufcrlct^t, muss in 
eoiisei'virenden Sinne wirkcn. 

flier i^reift nun dcr Wald wie keine anderc Vegetationsdcckc wirk- 
sam cin, thcilt durch scin Wiirrjclsystcm und seine Streiidccke u. durch die 
hiebei iniplicite geschaffenen tauscndfaciien uiechanisehen Hindernissc die 
abfliessenden Wasscrfadcn und zwingt sie durch l'>schopfung Hirer Kraft 
zur UnschadlicJikeit. Knnstnbildiing, Anrisse, AbsehiveDiDiinig der Feinerde, 
Aiisivasehung der VerzvitterungsseJiieJit, kurz jede Art von Terrainangriff 
mit all den nachtheiligen Folgen werden durch sein l^or/iandensein 
vcrniieden und die Uniwandlung von normalen BiicJien in WildbiicJie 
mit ihrer verderblichen Geschiebefiihrung und den gefiihrlieJien Regleit — 
und Folgeerscheinungen verliindert. 

Unumstosslich ist durch die direkte Beobachtung und lange Erfcihrung 
bewiesen, dass der Zerstorung des Bergivaldes die W'rtroekuung, I'erodnng 
der Hiinge und ini Weitfren unter deni lunfluss dcr meteorischen Xieder- 
schlage die rapid rjune/nnende Abfulir der Bodenkrinnme bis zum nakten 
Fehe>i folgt. Damit entfallen in erster Linie die 5<.V/('er\vassermengen fur 
die c2//t7/67zspeisung und gleichen Schrittes mit der Deterioration des 
Terrains ist die Versiegung dieser fi'ir den HausJialt und die Beted s sent fig 
von Aekergrfinden ete so notJiigen und liucJitigen Feuelitigkeitsspender zu 
konstatieren. Das fallt umso scinverer ins Gewicht als die oberfliichlich 
liegenden und durcli kein so niederse/ilagsreie/ies Einsiekerterrain unter- 
stittrjten Quel/en der tieferen Lagen erfahrungsgemiiss wiihrend der trockenen 
Jahreszeit ihre Thiitigkeit erheblich verringern oder gar einstellen. wenn 
in ^\c\\ tieferen Dage etwa tier Wald fehlen sollte. 

Die unschlid lichen l^ergbache tier beicaldeten Kpoche erleitlen eine 
sueeessive I'era/iderung iu troekene \\\\\\\s^\c welclie nur zu Zeiten heftiger 
Rcgen Wasser fuhren. tlann aber in geica/tiger Jlasse, da die zcitlich 
und rilunilich verzogerte Zufuhr tier l'\uchtigkeit zu ihnen \on tlen Hangen 
naek tleni I'utergange (/es W'aldes, /Proportional zum Wftcilderungs undl'er- 
kaklungsproeess Hires Finzugsterrains, inipetuos initl kurz dauer/td wurtle. 

Diese LImwandlung zu eeliten Wildbiichen mit immenser Geschiebe- 
fi'ihruug tritt naturgemass zuerst im (^berlaufe aller I'liisse auf, bleibt so 



i 



364 \h. IklVlc: 

laiio-c (/oriscibst bis rjii ciinin gL"iK.'i<iScn (h'adc localisirl , bis tier ] Fauptab- 
llusskanal ini (icbii\L;c niit all scincn Scitcnbachcn tlurch sciiic crodircndc 
Thatigkcit, ah Folgc dcr grosscii iiTCi^uliircii Wasscr mid CcscJiicbcfithntng 
iniincr iccitcr unci i^'citcr die seincn Lauf bci(]citcndcn l^Linhilngc iintl Ufer 
in illinlichcni Masse bcvii'jlusst , wie ilie fein.sten Riinsen es erstaials auftleni 
ehenialimn Waldterraiii tliaten. 

Vfcraid^rfichc, IlaiigLinsti'irrsc, kiir.:: ii.'cito-chcinh' WT'a'iisliiugc'ii diirch 
L' lit crw a seining uiid Corrosion fiihren inniier iiic/ir (iesehiebe und ]'\'ls 
triinimer deni Mittel- und Unterlaufe zu, bis endlich die Stini/f und 
Trniinncr'x'cllc die Mbene erreicht, und der an sich \iel wenii^er schadlichen 
Uberschweniniung des Landes die tjbcrlagcriing niit Schuttniassen hinrju- 

gCSc-IIt. 

Die Zeitdauer bis dieses Stadinni erreielit ist, bei deni die Aufnierksani- 
keit der InK'o/iiwr dcr libcno auf die unaufh.iltsani progressive katastropliale 
Gestaltuns^ der Wasserfi'ihruni;' t^elenkt wird, hani^t nati-irlich ab von der 
(irosse des rj'nzugsgebietes und namentlich von dessen goo/ogisi'/wr 
r^orniation. 

Weiche Sc/iicfcr iind JfcrgclgL-birgL i)(\c\- sandii;e / 'criciltiriiiigssi/iii/itcii 
untcrniischt mit ij;r6beren Gesteinstriinniien, i^^eben niclit selten Veranlassun^;" 
zuni Nietlergantj^ einer "' Mil/ire^ Jlier niengt sich das rasch abfliessende 
Xiederschlai;s\vasser so stark niit deni leicht abscliweninibareii Detritus 
bei kurzen aber lieftiiien (iew itterregen (Wolkenbri-ichen), ilass ein lava- 
ai'tii^er IJrei statt eines W'asserlaufes zu Thale eilt. Dicsc spc':::ifisc/i 
sc/iiv'crcrc Masse besitzt ein potcn:::irtcs ^1 ngrif/'s-und Trairf'ortvcrnidgcn 
fiir loses (iestin jcdcr (irbssr xoni haus_L;rossen i'V-lssti'ick l)is zuni kleinen 
Kiesel und die Ver\vi.istunL;en des ihren Lauf einsaunienden Terrains sind 
entsprechend i^esteiijerte, abgesehen tkuoii, class /cein H'assrr ini Standc 
7iV>Vt, so/ciw Steinnieui^en naeli Zalil und bes. Grosse in glciclicr Zcit" 
zu Tliale zu fiu'deil. 

'I'reffen solche elenuiitare Ciewalteii niit ihrein iMideffekt in bcwohntc 
Cicgcnden so ist die Wrici'istiing \i^\\ l^igenthuniw ertlien und die (n'Jalirdiing 
von ^Ienschenlel)en ini aus_L;"iebi!.;sten ATaase zu fiircliten. 

* Dciiu)iit/cy Ijcritlilc't iiiis. von ciiRr soIlIicii ^IuIic, wclclic \\\ ciium Viaw^c mil 05000 cinii 
\Va>scr iiicht wciiigcr tleim i6<;ooo cbtii. fcstc Masse /.u 'I'lial biachtc. 



\V;il<l [iihI >Vii>;s( r>vir1scli;in. 



365 



Die Scliuc'iz Tirf)] uiid l-'rankrcicli wciscn £Tcmi_c( vcruiistctc Plat7.c 
an dcii Mihidiiiigcn soldier W'ildicasscr auf, dcr Ortschaftcn siiul nicht 
wcnij^c, wclclic aiis dcni t^Icichcn Griindc vcrlasscii wcrdcn musstcn I 

In i^^cmiissi'ot kiihlcrcn Klimatcn odcr in (\cn warmcrcn, wo die ]*'Jcvatioii 
(kr (icbiroc cine i^cnui^ciu! i^rossc ist, fiillt die \'crlan[^sauiun<^ dtr 
Sc/i/ic'csc/ni/cI-Jc' diircli W'aldiicstocknnc;' cbcnfalls bcdcutcnd ins Cicwiclit ; 
die }iuo-cJicurcii I lochwasser aus naktcm 'J'crrain hci rnsc/u'r Sc/iii/csc/uiiclzi' 
iiiid die CJcfahrcu dcr Lawiiicu untcrstiitzcn ^\cn aus dcm vorii^cn uohl sclioii 
L;"cnutrend i^ercchtfertigtcn AuspnicJi auf Inzca/dinig des In-rglan Ics. 

Ich dcnkc, dcr Hinwcis auf die ludgcii dcr Entzcaldiuig rciclit Iiin. uni 
das Mittcl VAX zci^cn, wic all dicsen Ubclstilndcn bc^jci^nLt WLrden kanii. 
wcnn audi die Ztrst(')run^- Iciclitcr war als die \\'iede^llerstellunL,^ 

Man iiniss im jedeni />crg/gcj/ Landc, ckis cine gcsuiidc und prodiiktivc 
Wasscricirtschaft'ww Intercsse \on Ackerbau Industrie uml Landwirtscliaft 
aufreclit erlialteii will, eine entsprecbende Pn^haudhing ii)id Dcwirtschaftung 
f/cs Wn/dcs -d])^ f/iiid(7i/icii/n/c l-ordcniiig fiir die Jirrcicf/iiiig dieses Zicles 
verlaiigoi. 

Wo (lev W'ald ini Ikr^- u. I li'i^elland auf seinem nati'irlichen Standorte 
durcli lM'n<4ritT iles Menschen verschwand und \X'rschlechtert wurde. muss 
die schleuiiigstc W'icdcranfforstiiiig'xw weitmiM^lichsteni L'nifanj^e liLfurwortet 
werden. icciiii sic// iiac/ii^'eisoi /ass/, dtrss ]'crsc/i:ciiiden von IWi/d und 
/oca/c Storiingoi c/r/' W'asserfiihruni;- \"on Ix'ichen und kdiissen ini ursiichlichen 
Zusammenlian_<;"e stelien. 

1 )abei sollte wohl <;"edacht werden. (k\ss eine IV-deckuuj;" niit //V/A/ 
sc/i/cc/il/iin nic/tt !^enut;t, uni die entsprechemle Schut/u irkun;4 auf das 
Ti-rrain auszuiiben ; sc/t/cc/ttc W'aUlbestockun!:;" hat eben aucli nur teilweisc 
die L;unstii;'en Wirkuni^en zu \er/A'iohnen, w elelie dem L;ut L;».sclilosseneii 
und j^epfle^ten W'alde zukonum'U. 

Alle Manipulationen und \eben-Xut7.uni;en. welche (\c\\ \\ aid in seinem 
(lediilun beintriichti;;en. siiul auch a/s eine Inintriielitignug seiner Wirkung 
auf^^ufiissen. Vlu-rnit'Jssigc und un-eerni'inftige U'eide hat in l-'rankreich wie 
in Italien. Tirol u\u\ der Seluveiz (]cu Heri^wald herabi^ehrac'it und tlamit 
aueh seine wohlthatii^e Kraft nieht zuni kleinsten Theile i/lusorisr/i 
L;eniacht. 1 >ie Sc/int::en/dgesef::get>nng,\\v\c\\v eine Xutzuni;- ties W ?'■'.< 



366 Dr. Hefele: 

bcscJirdiikt in soldi en Lagcn odcr dirckt vcrbictct nnd anf ivc/c/ic rj. B. audi 
in Japan mit solchen Stolzc hingewicsen wird, hat nnr eincn Werth, wenn 
sie sich auch auf einen wirklicltcn ]]\i[ld unci nicht auf das Zcrrbild eincs 
solchen bczieht und notabene auch dor Willc und die Organc x'orhanden 
si nd, mn seine richtigc Behandlung zu liberwachen. 

Es ist nicht die Xutrjung ah solchc in der Regel, welche Nachtheile 
zeitigt, sondern die Art der XiitrjJing. J^on dicscni Standpunkt ist auch 
die Bewirtschaftung soldier W'aldungen zu rcgehi. 

KaJilscJilage sind namentlich in liohen und steilen Lagen unter alien 
Umstanden zu vermeiden ; ein rcgclloscr Pltxntcrbctricb mit seiner 
schleclit verdeckten Ausschlachtung und Verwahrlosung verdient eben so 
wenig <\c\\ Xanicn einer Wirtscliaft wenn er gar noch wie hierzulande, 
oft It liter dcni Dcckviantcl finer nnti'irliehen ^^er/'iingnng, besser gesngt 
Vericiistnng, segelt. 

Der Sannihieb mag auch hier wohl die besten Dienste leisten, wenn die 
Aufforstung iJnn auf deni Fnsse folgt. 

Wo die Wiederanff or stung nicht umgangen werden kanii, wird man 
zu imterscheiden haben rju'ise/ien nonnalevi Terrain, (lessen Wieder- 
bestockung ohnc weiteres eine mechanische Manipulation der kiinstlichen 
Holzzucht ist, und zwischen eineni ditreh die AtniospJiiirilien r^erstbrteni 
Terrain, dessen AbnorinitiU in Frankreie/i dein k/assiselien Lande soldier 
X'erwiistungen, eine eigenartige liochst wirksame Technik zeitigte. 

Nach dem Vorbilde Frankrcichs bcziiglich der Wildbachvcrbauung und 
Wiedcraufforstung der Gebirge haben Schwciz, 'I'irol, Bayern und Ostcrreieli 
seit langen Jahren denselben oder iUinlichen Wegen folgend, lu-f.ihrungen 
gesammdt, welche man gegebencn Falles sich unter alien Umstanden zu 
Xutzcn machen solltc. 

Auch in dieser }Iinsiclit ni()chte //V;' Japan, das ja erst am Anfange einer 
derartigen TItatigkeit steht, gar Manches zu lernen sein. 

Die Notwentligkeit zu energischevi Vorgeheni wird kaum melir bezweifelt 
werden, angesielits der W-rwiistungen seiner Jdiisse, :>.'e/e/ien alle Jahre 
eine stattliche .Anzahl von Millionen \'en (lOi an Werth, zum Opfer fallen, 
und abgesehen vnn deni dauernden X'erluste an k'ulturfahigem Boden in den 
jV/i'/ndunp'st/ia/ern tkr Fliisse. 



W'ahl und Wsisserwirtscliaft. ^67 

Mehr wic soiistwo hiingt hicr die Ik-bauunj^ dcs Landcs (Rcisbauy von 
der gcrc\c,^cltcn Versorgung mit Wasscr ab und cbcnso vichr z.'ii- sonstwo 
hindert dcr Wildbachchayaktcr die Aufschlicssuiig dcr Innenlandcs 
iiincrhalb oewisscr (Jrcnzcn durch dcii billigcn und cinfachcn W'asstr- 
verkehrsweg. 

Die Gcscltichcfiiltriiiii^r und das bcstaudigc Schi.'ankru zwischen Wasscr- 
losigkcit Oder dock schr nicdrigcn Wasscrstandcn und rdssnidni 
//<?^/m'^^^rr;/, in der Hauptsache zuriick zu fuhren :mU\\c fc/iicndc rir/itigc 
Bchandlung odc-y gar Verwii stung <!&<-, Waldcs und dcs Terrains \x^^\nn^:x^^ 
der Berglandschaft, sic liabcn cinen erhcblichen Antheil an dcr gcriugcn 
Rente schr wohl iiutzbarcr Wiilder, da das Holz auf i\<:n Wasserliiufen nicht 
in rieJitiger Weisc gebracht werden kann. 

Was hat man von der Zukunft zu crwarten, uciui man bei Fahrtcn. 
der Ost-Kiiste der Hauptinsel Hondo entlang. .svV///. wie die Kiscnbahnen 
auf endlosen eisernen l^riicken die enormen und in keinem Verhiiltniss zu 
ihrcr Wasserfiihrung erweiterten Scluittbette (h'verscr Wildwasscrflussc wie 
(Oigawa, Tcnriugawa. Fujigawa etc iiberquert, cnler gar. wie zwischen 
Osaka und Kobe mehrercmale unier den Fhissbetten hindurch fahrt. da 
die enormen Schuttkegel die oberirdische Fiihrung der Linie verbietcn. 

Die l^indammung dcr Fliisse auf ihren Schuttkcgchi. wic >:. W. dcs 
Minatogawa, der dureh Hiogo in ciner llr.hc v..n mchreren Mctcrn 
iUwr dcm Xivcan dcr Strassen bci Regenzciten seine schuttbeschwcrtcn 
Wogcn walzt. ist cin allerdings nicht mchr zu andcrndes abcr gcfahrh'ches. 
verzwcifeltes letztcs Hilfsmittel. dcnn cin einzigcr Dammbruch ist im Standc. 
unsiighches Ungliick zu vollbringen. 

W n-d hier nicht die Zufuhr der (Icschicbc durch AulVorstung unii durch 
Vcrbauung dcr ZuHiisse im JVrgland untcrbunden. so kann dcr Fhiss sich 
nicmals in scincn Schultkcgcl cinsclmcidcn nwO, so sclbst seine (^.cfahrlichkeit 
vcrnuiulcrn. sondcrn die fortihuicrndc AuriuMung ilcs I'hissbcttes bcdingt ih'c 
corrcsponihrcndc- h'.rhr.hung tk r Paninie be/ dieseui, x,'/r fid ydcm ttudnut/ 
W'i/dwasserjluss, uml mit iKr I'.rht.hung dcr Schutzwchr<.Mi wachst in 
Potcnzcn die Kataslrophc bcim Hruch.- ilcrsclbcn. /// n :s- 

urrt/u-ster \\\ isc- /st bereits all deni l.twiUtnlen in Japaii 
nierksamkeit :;>n .r.usiandiger Sei/e :ug<Wtndet i.'orden nnu .> >mJ 



368 Dr. Hofolo: 

Wildbachverbaunngen und Wiedcraufforstungen 7'ol/io- zerstorter Hiingc im 
iMiigugsgebietc diverscr Wildwasser im Gangc. 

Was mail dabci vcrmisst, ist dcr audi in aiidcrcii Spartcii so pcinlicli 
sicli gcltend machcnde 3[aiig-cl grosscn Zngcs. 

Ich \\urdif;"e liiebci schr wolil den Umstand, dass die i'lbrigcns in abseli- 
barer Zeit dcnnoch nolens voleiis sich von sclbst mclir u. mclir aufdrilngendc 
Inangriffnalinie soldier M'icdcraiifforstungcii iind ]^crbauuiigcn ini grosscn 
Stil, namliafte Millionen <in Gdclmittcln crfordern wird, die zur Zeit niclit 
bcschafft werden k()nnen, aber die gcsct::h'chcn Grundlagcn, die Fragc nacli 
dcr Bcitragslcistnng dcr betheiligten und Intercssenten nacli der l-5esdirank- 
ung dcs iMgenthumsredites, I^xpropiationcn etc etc kurzum die breitcrc 
J-5asis, die .\nbalinung" der Wcgc fur das cinJicitlichc Vorgehen und fiir die 
definitive unaufJialtsanie conscqiientc Aiisfi'iJiriing diescr ;/;//- vom Staat 
durcltfi'tJirbarcn ulrbcitcn miissten meines Kraclitcns eine baldinogliclisic 
Wurdigung erfahren, wenn nicht zusammehanglose Stiickarbeit das Rcsultat 
sein soil. 

Die Ausscheidung jcner Fliiclicn in Japan, welelie ivicdcr bcicaldet 
tverden iiiitsseii, diinkt micli cine der nidhigstcn Aufgaben der naclisten Zeit 
fiir planmiissiges \'orgelien, ebenso wie die Dispositionen fi'ir das in crster 
Linie dringend Xottcendigc und das ctwa AufseJiiebbarc und die Sorge fi'ir 
Beistclhing von ]\Iittcln. 

Die [''rage wird zweifellos hier nocli verwickelter durcli den Umstand, 
dass ein 'J'lieil der Ham unbedingt in diese Spiiiire ei/d'c.coge/! i\.'crdcni 
muss, nilnilich da wo die cnornie Stcilhcit der 1 li'inge die ,'\bfuhr der 
mcteorisclien Xiedersdililge in einer fiir das 'I'errain gcfaJirdroJicnden 
W^eise beschleunigt. 

I/ara und Rcisbaii sc/icinen aber V(^rerst in unaufliisbarer Wrbindung 
zu sein und doc/i kann bci niiJiereni Zuselien eine Landwirtscliaft nidit 
pr<)(Iukti\' grnannt werden, die direkt sich die Diingstoffc vom Walde und 
di r Ilara ohne eigentliclie Gegenleistung holt und indirekt spiiter durch 
die daraus \i:'r^\\\\.'\v\\\\(\c ZcrstoruJig des Waldes und scJilicsslicJi des Tcrranis 
d( ni I, audi- Millioneii kostet. 

I'",s ist eiiu- konstatirte 'riiatsadu-. dass die Beans f->rncliung der Ilara 
resp. ihres Grascs fiir die Ivi-iskuUur das ieir/:/ieI/e lieiliirfniss nanihaft 



WiiM iiimI >\ass['iMvir(soli;in. 369 

libersclircitct ; vicllculil, und icli zwcifle nicht daraii, wurdc schon cine 
Rc.liildion auf das Xothigc die A usschcidinig dcr gcfahrlichstcn Platzc ::ur 
W'lcdc-raiifforstiiiig ernioi^diclicii. Die Untcrsucluini(, in wic wcit cine anderc 
Diino-iingsarl in dcr Laiidwirtscliaft IMatz L^rcifcn kaiin imd leclchc Tlicilc 
von Ifara ciwa fiir die Uniformino- in spiitere Wcidcplatrjc i)assend sind, 
weiin eiiinial die- docii koninieiule VichrsucJit einen j^M-osscrcn Umfant; 
ant^eiiomnieii liat, das siml !•" ra<^a-n, dcrcn Ik-aiituortung so reelit eii,rentlicli 
in das (u-bict ciiicr landwirtscliafh'chen Akademie fallen wiirden. 

leli lialte das Sclieitern der vielen scithcrii,ren WtsucIic zur Jlebimi,^ 
der Vieii/aiclit, welclie tleni Ackerban eine ratioiicllere Dimgcrt|iielle liefcni 
wui-ck-, nicht VA\m gcriiigstcn r/icilc iladurch veranhisst, class man dcr Basis 
fill- dcrcii naliirlichc Fortcntwicklnitg d. li. dcr l^-ai^^c nach Wcidctcrrain. 
dcsscn Bcschafrenlieit. ni()olic]ic Ausdchun<^^ UnnvandkuiL,^ von Mara in .m,1- 
clics etc. etc niclit die nothlgc fioidamcntalc Aufiiicrksaiiikcit i^cschcnkt hat. 

Dcr Flcisclikonsum ist steioend und saniit die licrcchtit,ani£,^ /air 
liiufiihniiiq- //. Eniwick/mio- von Vich:zitchl t^cochen. 

Kehren wir zuriick zu unscrni ci-cnth"chcn 'Idienia unil koninicn wir 
zuni Schkisse ! 

WaldwwiX gcrcgcltc ]ldsscrvfr/iii/tnissc\ w ic sic die Landicirtschaft fiir 
cine -utc Ih-wiisscrnng dcs Landcs inul w ic sic llandcl Gtwcrih- und 
Industrie als Iransport und Kraft ntit Id \crlanocn. stclicn /// cngstcr 
Jk'zichuni;-, wic ich Iioffc ckarocthan zu haben. 

Wi-niag dcr Wald auch auf Ahwoxxwc Xicdcrscldai^svcrh.iltnissc unvl 
ckrcn /■a/gcn, die /foc/r.easscr, einen nnircrsa/cn und ///;• icrsagcndtN 
I'a'nlkiss uic/U auszuiibcn, so oibt dock oerack- HcwakJun- ein Mittel zur 
;v>7/4'c/H)urclifuhrun- dcs k:innusscs, iXcn dcr Mcnsch ulhr/iaitptaitfWW-^c 
Naturi^abc auszuiibcn \crniai;-. 

(Icradc fur japan, aui' ik-sscn Iiukistriezunahnic cin j;ut 'I'hcil seiner 
ZukunftscntwickkniL;- beruht. ist eh'c I IcrbciluhrunL;- einer ////(< '■//V//.n7 rcgc/niiis- 
s/gcn Wasscrfuhrun- seiner I'kissc unisonichr \ou cinschneidcnikMU 
Intcressc, ihx dcrcn natiirliclic Ik-seharfenhcit iknvh che Hcsonderheitcn 
dcs Terrains kcincs\vci;s cine an sick gitnstigc genannt werden kann. 

Die Ausniitzung dcr Wasserkraft in dcr Industrie ist zur Zcit cine 
mininiaic, uno-cfahr SoeXi rrcrdckraftc in aUcni. 



3/0 Dr. Hefele: 

Bcivaldung des Einzugsterrains ini Gebirge allcin vermag eine bessere 
Wasserregelung namentlich, wo die Flusslaufe durch laiige V'ernachlassigung 
cine weitgelitnde Verwilderung erfahreii haben, iiiclit ohne ii'citcrcs zu 
garantiren, cs nii'issen in den Biichcii mid F/fissoi srlbst aucli die ent- 
sprechenden teclinischen Wasser-Bautcn coryccl niid tccJinisch richtig 
aiisgefulirt werden durch Krriclitung von Thahpcrrcii, SauimckveiJicrn, 
UfcrvcrsicJicruHL^cii etc, urn mit dcin Waldc die so verderblichc Gcscliiche- 
fiihning, va\ der speziell die japanischen Fliisse infolge tektonischer 
Verhiiltnisse neigen, hintanrjitlialtoi. 

JJie wassertechnischen Bauten sind also als uiiuinganglicJi notJiige 
Stiitrjcn der Waldivirkuiig zii erachten ! 

Waln'end in den ( )berl;iufen niit den starken (jefiillen der Hauptwerth 
auf Vcrthcilinig iiiid Vcrrjogcruiig dcs Wasscrs rjii legen ist, bezwecken 
die Flusskorrektioneii, Dciclibaiitcii etc ini Uiitcrlaufc die raschcre 
AbfiiJirung der rjuJiiesseJideJ/ Wassermengen. 

l^as eintrilclitige Zusannnenwirken des Teelinikers und des Forstmatiiis 
wird den gewi'insehten Effekt gleiehiniissigerer, unseliiidlieJier Wasserfiilw- 
iing zeitigen. 

M. H. Die Natur hat ewige an Weisheit nicht iibertroffene Gesetze zu 
ihrer (irundhige und Xichts geschielit von ungefiihr ! Sie hat die Quellen 
und Wasserliiufe zu Fiissen des Bergwa/des und niclit eines kaJilen Terrains 
gesetzt, iln- Fingerzeig spricht deni Denkenden deutlieJi genug. 

Unterstiitzen sie den haclnnann, wenn cr sic uberzeugen konnte, durch 
das (jewicht ilirer Auschauung in deni Bcstrcbcn, Klarheit iiber den Wertli 
ties Bergwaldes fiir ein geordnetes Wasserregimc zu verbreitcn. 

iJann wird nicht ausbleiben, dass sich der alte Spruch mit ciner 
allerdint/s etwas freien Anwenduii"" audi hier w'icderholt ! 



>Viil(l 1111(1 Wiis-ci'wirlscliiin. 



,,'A';i(jtov ni/y vdan' 



Bciiiilzlc Literalur : — \Vflici-. Anfgal)fii dcr Forslwirtschaft. 
Ncy : Dcr Wald iind die OucIIen. 

Fraucniiolz : licsscit- BtMi.ilzung dcs Wa.<?;cr.s uiid dcr Was^cr'aufc. 
Coaz : Lauinen. 

Koch: Das sclincHe Au-cliwcllcii dcr Gcbirc;s\vas«cr. 
Klicrmaycr : Kinllus-; dcr Walder auf I'odcnfeuchtigkcit etc etc. 
W'csc : I'lcr die Wasscrahciiahme in den Qucllcn etc. 
U'aiif^ : (irundriss dcr Wiidl)achvcrl)auun£j. 

,, licwcgungsgesclzc dos Wa=per?. 

Dcmonlzey : Wicdcrhewaldung der Crchirgc. 
e'c, etc. 



Ueber Entstehung und Vertheilung des Kamphers 
im Kampherbaume. 



VON 



Homi Shirasa-wa, Ringaku/mkushi. 
(mit Tal'eln XXI— XXIII.) 



I. Einleitung. 

Der Kampher verdankt seinen Ursprung dcm Sckret von Cinnamo- 
mum Camphora Nees. Diesc Pflanzc waclist in tropischen und subtro- 
pischen Liindern. In Japan konimt sie ungefllhr bis 36^ nordl. Breitc vor, 
besondcrs abcr an dcr IMcercskiiste ; Shikokii, Kiushiu, Izu, Suruga und Kii 
bieten ihr den bcsten Bodcn. In diescn gegcn Kiilte gut geschiitzten- 
Gegenden, kommt sie neben anderen inmiergriinen Laubholzern in ansehn- 
lichen Mengen vor. Leider gehcn init der immcr zunehmenden \"crmehrung 
des Kampherbedarfcs diese Bestiindc allniahlig zu Grunde ; die grossen 
priichtigen ?2xempkTre triflt man nur mchr in Tempelhainen. Manchesmal 
erreicht der Kampherbaum eine Hohe von 20 m. und einen Durchmesser vor 
iiber 2 m. In Formosa komnit cr in grossen Bestanden in den Urwaldcrn 
vor ; die Kamphergcwinnung ist dort eine bedeutende lunnahmequclle der 
Regierung 

Der Kampherbaum Nvachst auch im ostlichen China in Tschi King und 
Kiangsi aiif iler Insel Chusan, siitllich \on Shanghai, Itschung in dcr Provinz 
Ilupe und auch der Insel Mainan. In diesen Gcgcndcn kommt or in mehr 
oder weniger grossen IMassen vor. wiihrend Formosa als das produktivste 
Land in Kampher bctrachtet wird. 



374 Homi Shirasawa: 

Ueber die Ausfuhr voji Kamphcr unci Kamphcroel von Japan vom Jalire 
1888 bis 1901 gibt uns das Kaiserl. Jap. statistische Bureau folgendc 
Angabc : 

aus Japan ; aus Formosa ; 

Kampher, Kampheroel Kampher, Kamphcroel 

Kin Kin 

1888 4,555,469 1,149,299 

1889 4,971,849 867,414 

1890 4,463,881 778,902 

1 89 1 4,429,050 ^S2 1,537 

1892 3,064,005 699,836 

1893 2,487,485 444.184 

1894 2,071,378 427.251 

1895 2,238,386 516,792 

1896 1,617,660 558,858 

1897 2,608,242 1,094,910 

1898 2,434,028 684,037 

1899 2,758,625 1,100,226 

1900 3.280,715 450,973 

1901 4,165,757 1,561,970 

, I Kin ist ungefahr i cnglisches Pfund.) 
Da von fast alien Lilndern der Welt, Japan die Hauptkaniplierproduk- 
tion hat und der Kampherbaum hier so wcit verbreitet ist, diirfte es fiir 
Forsttecliniker und Botaniker spezicll in wisscnschaftlicher und prak- 
tischer Ilinsicht von grossteni Interesse mid von Wiclitigkeit sein, ueber 
Ilerkunft, l^ntwickelungswcise und Vcrtheilung des Sekretes, resj). des 
Kamphcrs etwas zu erfahren. Bei der Wiclitigkeit des Kamphers fiir die 
Technik und die Wcitercntwickelung seiner Produktion insbesonders, Aviinsclie 
ich mit diesem Gegenstande mich zu beschaftigcn. 

Was nun die Behaltcr des Sekretes in den rtlanzenorganen anbelangt, 
so schrieb zuerst deBary' in seiner ,, Vergleichenden Anatomie " : ,, Nach 
tlem ]>au sind ilie Sek-retbehiUter /ii unterscheiden : in scliliiuche tl. h. aus 

' IX' liiry, Vcrgleithciulc Anatomic S. 143,152 und 2oy. 



4.395.921 


5,761. 


3,174,206 


65,753- 


2,292,098 


15.954' 


3,198,740 


6,440. 


1,571,200 





{ ('Iter Kiilslcliiiii^- iiikI Vcitlu'iliiim' lU-s Kain|)iM'r> iiii KiiinpliiTlt.tiinic. 375 

Zcllcn, \\\lchc ihi'u W'andc l^eibchaltcn, hcrvor^cgangcnc, dahcr gcwolinlicli 
als Zcllcn ]jc/xicliiictc unci intcrccllularc, ihrcr Gcstalt nach cntwcdcr, 
(iilngc (xlcr Liickcn, I lohlcii zu ncniicndc." l^rstcrc untL-rscheidLt cr nach 
den I'ornicii in ,,kurz(j und langc'' und Ictztcrc in ,,schizogcnc" und 
,,lysig"cnc" ,,odcr rhcxigcnc". A. Tschirch' wclchcr sich in den Ictztcn 
zclm Jahrcn specie! 1 niit den Sekreteii der Pilanzen bescliiiftigt Iiat, nennt die 
oelfiihrenden Zellen ,,(Jelzellen"- ; die scliizogenen Li'icken ,,()el-oder I Farz- 
behalter" (audi Oelrauni; ; die lysigenen ,,Oel oder I larzlucken". Fiir die 
scliizogen cntstandenen und spilter aufl)'sigenc .Vrt erweitertcn Behalter hat 
er den Namen ,,Sc]n"z(j-l)'.sigene Raunie" eingefi'ihrt. ,,Obhtoschizogene 
(lilnge" liat er sic genannt, wenn die Sezernierungszelleii der Oelbchalter 
erst nacli Bildung des Iklages obh'terieren. 

Uebcr dicse Sekretbehalter, speciell ch'e k.nstehungsorte ties Sekretes, 
liaben in letzter Zeit Tschrich''' und dessen Scluder eingehende Stuch'cn 
geniaclit. Becheraz, ..Ueber die Sekretbildung in schizogenen (iangen". 
Bern 1893; Sieck, ,J)ie schizcjgeiien Sekretbehalter, xorneiinilich tropisciier 
J leiljiflanzen", l^ern 1894; Lutz, ,,die oblito-schizogenen Sekretbehalter 
tier ^lyrtacecn", Hern 1895 ; Bierman, ,,Ucber Ban und luitwickelungs- 
geschichte der Oelzellen und die Oelbildung in ihnen". 1898. 

Unter diesen Arbeiten hat Bierman ini Si)eciellen iiber Oelzellen und 
die Oelbiklung in ihnen bei den Ptkinzen von den Faniilicn der Lauraceen, 
(Cinnanioniuui Caniplu)ra ausgenonmien) Canellaceen. X'alerianaceen. Zin- 
giberaceen, ?^ragnoliaceen, MN'ristiaceen, l'ii)eraceen. Cal)'canthaceen. L ni- 
belliferae, Convol\-ulaceen. Papilionaccen, I'ntersuchungen angcstcllt. 

In gar nianclien cheniischen und j)harniazeutischen W erkon findcii wir 
Abhandlungen iiber den Kani[)her ; aber da die Ouellen zu dicscn nicistens 
dieselben sind, beschriinkcn sic sich auf seine cheniischen l\igenschaften und 
seine Anwendung. etc. Was aber die Mntstehung und X'ertheilung des 
Kaniphers ini Kanijiherbauni anbetritVt, so ist eine eingehende L'ntcrsuch- 
ung, besoiulers die niikr(\skopische h\>rschung. noch niclit bekannt. 

' I'scliiich, Anijt.\\an(lto ril.ui/on Au.uonuV- S. 41x3 530. 

-' A. Meyer liat tViihcr uicscii Naincii ciuy[ofu!ut : \Vissen-;chafilich l)r«>i:;cnkunik' 1S3, S. 79. 

* Tschircli, liar/ uml Ilarzbchalter, kxx). 

Ti-chiicli nil J Oostcrlo, Anatomisclior AUa-^ lic" l'l»annakoijiK>sic uiul naluun!;r^mittclkuiKlc, 1901. 



3/6 Homi Sliirasawa : 

Fliicki<4"cr ■ schrcibt : ,,dcr Kampher findct sicli auskrystallisicrt in 
Spaltcii dcs Stamnics. sowic aufgclost in dem Oel, welches in alien Theilen 
des Baumcs (mit Ansnahmc dor Bliiten) verbreitet ist. Pfeffer^ hat die 
Ansicht, dass das aetherisches Oel der lebcnden Zellen des Kampherbaumes 
dufch cine postmortale SauerstofRiufnahnic ini Kampher verwaudclt wird. 
In W'eisnicr's Buch steht^ ;" allc Theilc des Baumes cnthalten in besonderen 
Sckrctzellen cin aetherisches Oel, aus welchem zuni Theil schon in der 
lebenden Bflanzt: der Kampher im krystallinischen Zustande sich aus- 
scheidet. 

In Bezug- auf den (iehalt von Kampher im Kampherbaum habcn 
neuerdings Moriya^- und ^^limura durch Dcstillation von IIolz und Blattern 
werthvolle Ergebnisse, besonders mit Rucksicht auf Kamphergewinnung 
gehabt. Sic schreibcn : ,,der Kamphergehalt im Kampherbaum ist sehr 
verschieden je nach dem Alter und Standorte des Baumes. Bei demselben 
Exemplar nimmt er vom W'urzelstock nach den Aesten hin ab. Das 
Kernholz enthalt mehr Kampher als das Splintholz. i\us dem aus Formosa 
stammenden ungefahr 500 jidirigen llolz haben sic 7.13 proz. Rohkamphcr 
gcwonnen, wahrend sic aus dem 70 Jilhrigen Holz, welches in der Provinz 
Bingo gewachsen ist, nur 4,73 jm-oz. Rohkamphcr gcwonnen haben. Die 
lufttrockcnen Blatter cnthalten tlurchschmitlich 3.75 ]m-oz. Rohkamphcr und 
1,25 proz. Kami)hcroel. 

Ucber den Kamphergehalt in den Blattern von Cinnamomum Cam- 
phora theilt uns AI. Kelway Bamber"' im Royal Botanical tiarden, Ceylon 
mit, tlass er durch Dcstillation von lufttrockcnen Bliittcrn durchschnittlich 
2,oSo bis 2,425 [M'oz. Kampher und 1,96 bis 1,05 [)roz. Kamphcroel gcwon- 
nen hat. 

In meincr \'orlicgcndcn Arbeit w ird es sich haupts>lchlich tlaruui haiidcln, 
nach iiiiki-oskcpischtr L' ntcrsuchuiig folgentle I'^-agen zu bcantworten. 



' rliafiuicji^iiusic des i'll.iii/.ciirciclics. iS^iiS. 150. 

'' I'(Lituc-n[)liysiii!(iL;ic I. 1 AuT. S. 501. 

•' Die Rolistoflc (Ics rihui/.cnrciclics kjoo. 

* lounial of tlic dicinical Society Tokyo, \'i)l. 21 Nd. uimI Uiiij^akusliikwai Z.isslii^ll. 

"• 'I'lic Imlian l'"orcstcr \t>\. 28 No. 4, iyo2 S. iGj. 



{ olior Entsteliiiiig iiiul VoHIumIiih!? dos Kamphors iiii Kaiiipiu'rbniiiiic. 377 

T. Alls wclclu'i- Sul)stanz sclicidct sicli dry Kamjihcr im Kanii)li«rbaiini 
aus ? 

2. Wann maclil sicli das crstr I'lodiikl, aiis dcin sicli dcr Kamphcr 
l)ildct, bci dcr wacliscndcn I'flanzc jjcnicrkbar ? 

3. In wclclu" l-'orm t4(-ht das zucrst cntstaiukne I'rodiikt scciiiuliir 
i'il)cr, iind was sind seine mikroskopischcn I^it^cnschaftcn ? 

4. \\'(i wii'd das crstc I'rfxlukt ange]t;L;"t und wodurcli cliaraktcrisicrt 
cs sich ? 

5. \\^o vcrtliL'ilt cs sicli in ckn Tflanzcnort^ancn und in wclchcr l^.Lzicliiin^ 
zu kliinatischcn und sonstii^cn \"crhiiltnisscn stcht seine ]^ilduni(? 

6. Kann man voni Standpunkte dcr Kampherocwinnunf^ aus, den Kam- 
pheroelialt im Baume miki'f)skopiscli bcstimmen ? 

iJicse I'litersucluinoen wurdcn im Jalirc 1900-1901 in der forstliclien 
Versuclisanstalt in Munclu n und hauptsiiclilicli im Jalire 1901-1902 im 
l)liarmazeutischen Institut der Universitat l^ern und spiiter im rorstlichcn 
VtM'suclis^arten in ]\Teo"uro bci Tokio, austi'efulirt. 

Das K'l)ende \"ersuclismaterial stammte aus dem botanisclien Garten in 
Aliinclien, und Hern s(nvie audi aus ilem fdrstliclien \'ersuclisj;arten in 
Abi^uro bei 'l'()l<i(^ ; das tiockene ^Material aus der SammluiiL;' des llerrn 
Trof. Dr. A. Tscliircli und aus meiner eit;\nen. I brr Prof. Dr. W. Alayr 
iiberreiciite mir auch das trockcne Material aus der SammluiiLi'. die er auf 
sciiur keise in ja])an aiiL;elei;t liatte. Mein College I lerr a. o. Prof. 
ATimura, Tokio liat mir \'erschiedenes I lolzmateriai aus der Prcn-inz I'ukuoka 
besorj^t. 

]">s ist mir cine anL;"enehme Pllicht diesen llerrn. vor Alleni llerrii 
I'rof. Dr. A. Tsclu'rcli. unter (lessen freundlicher Leitun*;" ich liauptsiiclilicli 
diese P^ntersucliun<;en im pliarmazeutischen Institut der Unix'crsitat Bern 
aust^'cfiilirt habe, dem inzw ischen \'erstorbenen llcrrn Prof. Dr. R. llarti^'. 
und llcrrn Prof. l)r. II. Ma}-r an der l^nixcrsitat Munchen, welche walireiui 
meines Aufenthaltes in Ariinchen fiir tliese Arbeit ihre t;-utio-e Ililfc mir 
i;"c\\iihrteii. meinen \\;irmstcn uiul \erbiiulliclistcn Dank auszusprcchcn. 

Tokio, Sc[-)tcmber i(^02. 



78 Komi Sliirasawa : 



0/ 



II. Entstehung. 

Kii(>s/>in. 

Um fcstzustelleii, wo sich tlie crstc Anlac^c der Oclzcllen vollzicht, und 
wo man ihrc friihcstc iMitwickcIungsg-cschiclitc wahrnchnicn kanii, habc ich 
zucrst an den Knospcn L'ntcrsucliunoen aus^cfiihrt. Dazu dientc mir 
frischcs AlatLi'ial aiis dcm Ijotanisclicn (Tartcn IVtii. Die diinnon \\\c audi 
die dickci'cn ScbnittL' in der (Hkt- uni\ LiinL;sriclitun<;' wurdon direct in etwas 
verdiinnte i proc. ( )sniium.silnre oder stark xerdiinntc Alkannatinktur t^'e- 
taucbt. In ersterer L()sun_L; ^\nrden (]ie Schnitte nacli einigcn Minuten. in 
letzterer nacli einiL;en Stunden untersuclit. IV-i nieinen nie]irniali;^"en Unter- 
snchnn;4en konnte icli tlie ( )elze]le oder eine tier letztern entsprechende 
Zelle niemals finden. Die flachke^elii^e Spitze wird \'oll.standi^" aus weichem 
mcristcmatisclien (iewcbc t^ebiklet. Schon direct liinter dcm Vcgcta- 
tionspunkte treten in uni'CLtelmassiL^er \'ertheilunL!;" ciriit^e Zcllcn auf. 
\\'elclie sich clnrcli (in'jsse und rundliche l-'orm \on den iibrii^'cn Zellcn 
auszeiclmen. Sie reai^ieren noch nicht mit Osmiumsaure oder Alkanma- 
tinktur, und der Zellinlialt ist etwas ckuxhsichti^'. l-5ctraclitet man dicse 
Zellen an dvn in Alkoliol lic<j[enden Praparaten so bemerkt man, dass ilire 
Zellrilume mit einer farblosen homogcncn Masse erfiillt sintk Dicse ?ilassc 
(juillt (lurch N'orsichtii^es Zufiicssenlasscn \on Wasser und schwindet in 
Alkoliol. Mit Jotljodkali nimmt sie eine hells^elbe Fiirbuni;' an. Dieses 
deutct auf Sclileini. -■ W ir wissen ja (Un-cli die Untersuchungen \-on 'I'schirch ' 
und l^iermann-, dass cHe Oclzcllen als Schleimzellcn an^elci^t werdcn. 
Unmittclbar hinter tlem V'egctationspunktc, bcsondcrs an den Randparticn 
der Knospenachse fmden sich die ijjanz diffcrcnziertcn Oclzcllen, wclche 
durcli (Jsmiumsiiure oder .Alkannatinktur nachc^cwicscn werdcn k()nncn, 
Fi- ,. 

Unter denselben machen sich alxr nocli anderi' Zellen bemerkbar, 

' Die liar/ 1111(1 llaizhcluilltr. 

* l'cl)cr I'au und F,nt\vicl<cluiigSL;cscliichiU' <lt.r ( Vl/cllcii. 



Ichcr Kiitsteliiiiii>- mid Vcrthciliiug^ dcr Kampliors iiii Kaiiiplicrbaiiinc. ;>79 

welchc in den vcrschicdcncn Statlien dcr Kntwickclung der OclzcUen stchcn. 
In das crstc Entwickclungsstadiun, also in die Zeit des Uebcrgchens der 
Schlcimzcllc zur Oelzellc, filllt das Auftreten dcr ,,resinogenen Shicht" 
Tscliirch's, — dicsc Schiclit stcllt cine feinkornige vacuolige ]\Iasse dar. In 
der Zellc ist die Wand mit ciner Sclilcimmcmbran dicht bclcgt, wiihrend 
die rcsinogene Schicht inwendig dersclben auflicgt, Fig. 2, a. 

Die von den verschiedenen Particn des Schlcimbelegcs licrausgehendcn. 
bandformigen Schlauclie treten in der Alittc des Zcllraumcs zusammen. 
Fig. 2, b. 

Im Zusammenhang mit dcr succcssiven Kntwickclung tier rcsinogencn 
Schicht stcht das allmahligc Vcrschmelzcn des Schleimbeleges. Es ist 
jcdoch audi moglich, dass diinnc Schichten des Schleimbeleges odcr Reste 
dessclbcn zuriickbleibcn. 

Das crste Auftreten von Oeltropfen konnte ich bald nach dem Ent- 
stehen der rcsinogencn Schicht feststellen. In Ictzterer fimden sich die 
Oeltropfen hilufig in der ]\Iitte, jcdoch auch in beliebigcn andcrcn Punkten. 
Dicsc Oeltropfen wachsen bald zu ansehnlicher Grosse hcran, was bewirkt, 
dass die bctrcffcnden Stellcn der rcsinogencn Schicht aufgcblasen werden 
und das (ianzc in den Zellraum hincintritt. Die aufgeblasenen Partien 
zeigcn manigfache Cicstaltungen T^ig. 3 — 7. 

Im weitcren Fortschritte der Oelbildung treten die genannten Partien 
miteinander in Contakt und erfiillen <.\cn ganzen Zellraum. lassen aber nicht 
selten Lumen in tier ^vlittc odar an der Zelhvand zuriick. Fig. 8 — 9. Dicsc 
Masse,— sammt dem in ihr bcfindlichen Ocl — ist ziemlich fest. Bci Unter- 
suchung dcr Knospcnachse bci trockenem Material konnte ich sie mit dem 
Messer zcrtheilen und ohne I'cM'nneriinderung aus der Zellc hcrausnehmen. 

In cinigen FiUlen habe ich im Zellraum ilic rcsinogene Schicht stark 
aufgcblasen gesehen, welchc dadurch ein beutelformigcs .\nsehn crhielt, 
P^ig. 10. Bringt man cine solchc Piltlung mit Alkohol in Pcriihrung, so 
schruuijilt sie baUl zusannnen. (^hne Hehandlung mit einen Rcagenz hat 
ilic obenerw idmte Masse ein hellgclbcs, homogencs ctwas lichtbrechemles 
odcr schaumigcs Aussehn. 

Osmiumsiiurc f.irbt diese Masse dunkclbraun uml kontrahicrt ctwas. 
Mit Alkannatinktur nimmt sie cine intensive rothe Farbunt: an. Durch die 



3^0 Hoiiil Shirasawa: 

Behandlung mit Alkohol dehnt sie sich aus unci es bleibt cine feinkornigc 
Masse zuriick. Zusatz \'on Wasscr hat zuniichst zur Folge, dass die Masse 
zu einem ovalcn oder rundlichem Gebildc kontrahiert wird, welches ein 
triibes Aussehii besitzt. 

Diesc Tatsache, welchc den allgemeinen Gesetzen entgegengesetzt zu 
scin scheint, liisst sich vielleicht auf folgende Art erkliiren. Bci Hinzufiigung 
von Alkohol tritt momentan Volumenvcrmehrung der schleimigen Masse 
durch die Losung des festes Oels ein. Setzt man Wasser hinzu, so wird 
infolge Quellung die Oellosung hcrausgeprcsst, und tritt einc Zusam- 
menziehung derselben ein. 

Durch fortdauernde Einwirkung \on Alkohol und Wasser wurde sie 
nach und nach klarcr und zuletzt blieb eine schwammigc, oft grobporige 
oder nctzartige etwas lichtbrechende Masse zuriick. Dicse Masse speichert 
schr begierig Anilinfarben auf. So filrbte sie sich mit Jodgrun-Eisessig 
leicht griin, audi nach Abwaschcn mit Alkohol oder Glycerin wurde die 
Fiirbung ganz energisch zuriick gehalten, wiihrend das andcre Gewcbe 
farblos blieb. Mit Jodgriin und Fuchsin fiirbte sic sich schon violet. In 
Chloralhydrat, cone. Schwefelsiiure oder Salpetersiiure ist die Masse unlos- 
lich, Chlorzinkjod gab ihr eine gclbe Fiirbung. 

Diese Rcactioncn berechtigen mich, sie als ,,rcsinogene Schicht" zu 
bctrachten, welchc Tschirch als das Eaboratorium der Oelbildung (Harz- 
bildung) bezeichnct hat. 

Was fiir cine Rolle diese resinogene Schicht in der Oelbildung spielt, 
ist noch ein Riitscl, wir miissen heutzutag noch der Kraft dcs lebenden 
Protj)lasmas die Fiihigkeit der Oelbildung zusprechen. Zur Zeit kann ich 
jedoch als sicher erkliiren, dass der Ort der Sekretbildung bei dem 
Kampherbaum, wie bei den anderen Lauraceen in Tschirch's resinogener 
Schichit zu suchen ist. 

Iki den Untersuchungen von Knospen habe ich bei der ersten und 
zweiten l^lattanlage, d. h. bei den innersten Schuppen nur Schlcimzellen 
bemerkt. ^\uf dem Liingsschnitte sind sie meistens am iiusscren Teile in 
verticalen Reihen augcordnet. In einigen {"'iillen aber waren bei der dritten 
Blattanlage die bereits ausgebildeten Oclzellen oder die Schleim/.ellen mit 



Ueber Eiitstehun^ iind Vertlieiliiiig dcs Kaiuphcrs im Kaiiipherbaiiine. 3'^i 

(Icr resinogenen Schicht bcmerkbar. Von hicr nach den ilusscrcn Blattan- 
lagen zu ist die Oelzellcnbildung cine intensive. 

Was die Entwickelungsgeschiclite der Oelbildung anbetrifft, so liisst sich 
ein wesentlicher Unterschied bei dicsen zwei beschriebenen Fallen nicht 
erkennen. 

lUattcr. 

Bei dem kaum entfalteten Bliittchen von etwa i ' \, — 2cni Gesammtlange 
sind bereits die Schleim- und Oelzellen vollstiindig ausgcbildet und zwar in 
Blattspreite und Blattsticl. 

Die Oelzellen sind mittelst Alkannatinktur oder Osmiumsl'iure nach- 
weisbar. Die Zahl der Oelzellen ist jcdoch meistenteils noch verhiiltniss- 
miissig gering, wahrend sich \ielniehr die Schleimzellen in grossercr Zahl 
benierkbar machen. 

Bei den einjahrigen l^Uittern, welche sich bereits v()llig entwickelt 
haben, weisen die Oelzellen schon ihre charakteristische Ausgestaltung und 
vollig entwickelte (jrosse auf. Die verschiedcnen Stadien der Oelbildung 
sind jedoch bei diesen nicht nachweisbar. Die Oelbildung in der resino- 
genen Schicht hat zur l^'olge, dass dieselbe stellenweise aufgeblasen wird 
und nach innen in den Zellrauni eindringt und zwar von der Seite der 
Zellwand, der die resinogene Schicht entweder unniittelbar angelagert oiler 
vor der sic durch eine Schleinimenibranschicht getreinit ist. Diese blasen- 
artigen (iebilde sehen rundlich, eifch'mige oder wurstf(')rmig aus. Fig. II. I2. 
13, 14, 15, 16. Manchmal simi eine grosse IMenge derselben beieinander 
angelegt und sie erfiillen dann den Zellrauni. Fig. 17. Ohne Zusatz eines 
Reagenz haben sie ein f.ist farbloses oder hellgelbes, stark lichtbrochendes 
Aussehn. Durch Zuiliessenlassen von Alkohol kontraliieren sic sich baki 
und bleibcn hierbei Fctzen der resinogenen Schicht zuriick. Dicscr Fctzcn 
des resinogenen llilutchens fiirbt sich niit ciner LCtsung von Jodgriineiscssig, 
alx'r die Fiirbung ist keine intensive, nut Jodjodkali ninunt cr cine orangcn- 
gclbc k'ilrbung an. 

Untcr tlen Sekretzellen dieser Hli'itter trifft man noch vicl Oelzellen, 
welche .uif ihrer Innenseite niit einer tlicken Schleininienibran ncbst der 



382 Homi Shirasawa : 

resignogencn Schicht xcrsehcn sind. Biswcikn crscheint die resinogenc 
Schicht beutelformig. Hiebei bliebt die farblose Membran durchsichtig und 
die resinogene Schicht sieht etwas tri'ib aus. Mit Jodjodkali fiirbt sich die 
erstere hellgclb. wiihrend die Ictztere eine orangegelbe Fiirbung annimmt, 
wodurch man das Vorhandensein der Schleimmembran und zugleich die 
AbgrenzAuig zwisclien den beiden leicht \\aln-nehmen kann. 

Bei den zweijahrigen BlLlttern sieht man an der Blattspreite ueberhaupt, 
und auch an dem einzehien Geweben z, B. Epidermis, Zelhvand, etc. eine 
bedeutend dickere und festere Struktur als bei den einjiihrigen Bliittern. 
Die Oelzellen sind bei zweijahrigen Bliittern nocli in den verschiedenen 
Stadien ihrer Entwickekmg. Sie zeigen noch meistens Schleimmembran 
oder die Reste desselbcn. obwohl dininer und weniger zahlreich als in 
einjiihrigen Bliittern. Tropfenweise vorhandenes Oel habe ich nur in 
wenigen Fiillen gefunden. Es bildet sich wie bei den einjiihrigen Bliittern 
in der schwammigen resinogenen Schicht, oder in blasenartigen Beutelchen. 
Ich konnte einen grossen Untcrschied in der lintwickelung nicht bemerken ; 
ein Unterschied besteht jedoch darin, dass in den zweijiihrigen weniger 
Sclileim, hingegen mehr Oel in der resinogenen Schicht vorhanden war als 
bei den einjiihrigen. Ich bin daher der .\nsicht, dass der Prozess der 
Oelbildung in den Bliittern c;in fortdauernder ist und zwar wiihrenti ihrer 
ganzen Lebensdauer. denn untcr den zu meinen Untersuchungen beniitzten 
zweijiihrigen Bliittern zeigten sich stets die gleichen Entwickelungserschei- 
nungcn, wiewohl einzelne der Bliitter schon die auf baldiges Abfallen 
hindeutende charakteristische mattgelbe Fiirbung hatten. Meist war 
entsprechend dem ICntwickelungsstatliuin der Bliitter ein successiver 
Fortschritt in der ( )e]l)ildung zu bemerken. 

Bei (leii Hlattstieleii steht das Fortschreiten der Oelbildung fast immer 
im entsprechenden Wrliiiltniss zu tlen zugeh(")rigen Bliittern. Bei tliesen 
habe ich wiederholt den trojifenformigen Oelinlialt in ilen (X-lzellen 
bemerkt, l'"ig. iS, uu<.\ zwar ist dtrscllie in zweijiilu-igen Bliittern reichlicher 
als in (.-injiihrigen. In mehreren h'iillen war abrr die resinogene Masse mit 
Oel durchtriinkt xorhaiidin. 



lelier Entstchiiiig mid VtMthpiliiiig' <Ios Kaiiiphcrs im Kiiiiipliorbaiiino. 3''^3 

Durch Bchandlung^ mit Alkohol losten sich die Ocltropfchen sehr bald, 
Aviihrend in den rcsinogcnen Schliiuchcn sich das Ocl erst unter daucrnder 
Einwirkung von Alkohol loste. Schleimmembran oder die Reste derselben 
bleiben zuriick, jcdoch fehlten sie audi manchinal, oder waren iiber- 
haupt diinncr luid gcringx-r als in den Blilttern. Die Blattsticle geben 
immer cin sehr zweckmiissiges Material fiir Untersuchung der Oelzellen, da 
Avir bei denselben je nach Belieben einen di'innen oder dicken Schnitt 
l)ekommen konnen und da auch wegen des geringen \"orhandenseins von 
gcfarbteni Zellinhalte im Gcwebe immer ein klares Bild wahrzunchmen ist. 

Bei der Untersuchung von BUlttern uml Blattstielen dcs trockenen 
Materiales aus Java stammend, habe ich auch Oel in tropfenformigem 
Zustande gesehen, Fig. 19. Auf Osmiumsiiure reagiert es schwach, abcr mit 
Alkannatinktur farbt es sich schon rot. In Alkohl l(")st es sich bald, unter 
Zuriicklassung \on nur wenigem Rvickstand. Reste der Schleimmembran 
waren bei dicsen kaum bemcrkbar. 

Auffallend ist, dass man oft in Knospenachsen, Blattstielen, Rinde. auch 
im Holz, netzformige oder hautartige rcsinogene Masse findet, welche fast 
die ganzen Zellriiume ausfiillt. Bei Bliittern fehlt dicsclbe in der Oelzelle 
fast immer und ist bei frischem Material durch dickc Schleimmembran 
ersctzt. Fiir tlen Nachweis der Verkorkung der Oelzellwand, auf welche 
Zacharias ' zuerst liingewiesen hat, habe ich SchwefelsLlure. Chlorzinkjod 
oder Jodjodkali benutzt. Ik-i den jiingeren Pflanzenorganen wurde durch 
cone. Schwefelsaure das ganze (jewebe zerstin't. wobei die feine Zell- 
wandlamelle tier Oelzellen zuruckblieb. Bei (\cn alteren Organen farbtc 
sich die Zellwand durch Jodiodkali orangegelb. 



/\ ///(/(■. 

Die von mir \orgenommenen I'ntersuchungen erstrecken sich aut 
frisches und trockenes Material, unil zwar auf die jiingsten Teile einjiihriger 
Triebe. 

' luilanisclic 'A-i1u!i;j; 1870. S. 617-645. 



;84 



Hoiiii Shirasawa. 



Beziiglich der Oelbildung treten fast die gleichcn Vcrhaltnissc hcrvor, 
wic bei den einjiihrigen Bliittern. 

Gelegentlich der Untersuchung der Rinde 5 jiihriger Triebe war zu 
constatiren, dass der Prozess der Oelbildung schon zum Abschluss ge- 
konimen war. Hellgelb aussehende oft scliaumige Oelmassen erfiillten das 
Zcllinnere. 

Durch Behandlung der Oeltropfeii mit Alkohol losen sich dieselben 
iinter Zuriicklassung einer f^irbloscii schwanimigen oder einer hautartigen 
^lasse 



Holr. 



Ueber die Kntwickelungsgeschiclite der Oelzelleii bezieliungsweise die 
Oelbildung in demselben erfidirt man nur weniges, dagegen kann man bei 
demsclben ausschliesslich die interessanten Umwandelungsformen des ent- 
standenes Oels zum krystallinischen Kamplier studieren. 

Tm Wasserprliparat des jungen 2, 3 (xler 5 jiihrigen Holzes sieht man 
nur i\Q\\ hellgelben Oelinhalt in der von den Nachbarzellen ausgezeichncten 
Oelzelle. Er erfiillt (Xcn Zellraum oder einen Tell desselben. 

Mit Alkannatinktur farbt das Oel sich rot, durch (^smiumsiiure wircl es 
gebriiunt Alkohol lost das Oel leiclit und bluiben manchmal mcmbran- 
artige oder schwannnige Klumpen zuriick. 

Beim ungcfahr 80 Jahre alten imd vor 12 Jahren gefiillten Holz, 
welches aus einer bekannten Kampherproduktionsprovinz stannnte, habe ich 
hauptsiichlich die Bildung vom Kampher untersucht. Wenn man diinne 
Schnitte dieses 1 lolzcs im W'asser betrachtet, so sieht man vier verschiedene 
I'^irmen : 

a. Dunkles oder orangegelbes C^tl 

b. hcllgelbcs Oel 

c. farbloscs Oel 

d. Krystalle. 

Das dunkle Oel ist eine balsamartige Masse ; bei tier mikros- 
kopischen Untersuchung tritt es oft aus der Zelle aus, wobci es seine 



Uebcr Entsteluiiij? wml Vertheilung' des Kaniphers iiii Kampherbaiiiuc. 3^5 

urspriinglichc Form bcibehiilt. Dassclbe hat meistens ein schaumigcs, 
triibcs Anselin, Fig. 20 und erfiillt fast immcr den ganzen Zellraum. Alkoho! 
lost cs langsam unter Bildung von zahlrcichen feinen Hohlraumcn. Durch 
andaucrndes Zuflicsscnlasscn von Alkohol nchmcn dicsc an Zahl und 
Grosse zu, und bleibt zuletzt cin schwammigcs netzartigcs Skclctt zuriick. 
Durch Aether oder Chloroform geht tlie Losung schneller vor sich. Mit 
Alkannatinktur fiirbt es sich intensiv rot, mit Osmiumsaure braun. Das 
hellgelbe Oel ist dagegen etwas durchsichtiger und schliesst meistens in 
der Mittc oder in den der Zellwand anliegcnden Partien grosse Luftblasen 
ein, Fig. 21. Alkohol lost es leichter als das vorige, bei der Losung bleibt 
ebenfalls ein Riickstand. 

Das farblose Oel ist cine ganz ckirchsichtige homogene sehr fliichtige 
Flussigkeit. Es kommt in der Zelle tropfenweise vor, und habe ich nur in 
zwei Fallen ganz mit demselben erfiillte Zellriiume 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 tretcn in dem Tropfen kleine Luftblasen auf Seine 
fliichtigen Eigenschaften kann man gut feststellcn. Unter dem Mikroskop 
betrachtet verfliichtigt es sich schon in einigen Minuten, wobei ein in 
Alkohol loslicher kleiner Klumpen zuriick bleibt. 

Nicht seltcn findet man in der Mittc dieser Oeltropfen oder am Rande 
derselben kleine Krystalle, Fig. 28. Bringt man dicse Krystalle oder die 
Tropfen selbst mit Wasser direct in Bcrlihrung, so drehen sic sich sehr 
lebhaft und vcrschwinden schliesslich vollstiindig. Dieses Oel farbt sich 
sehr schwach mit .Vlkannatinktur oder Osmiumsaure. In Alkohol oder 
Aether ist es sehr leicht l(')slich. 

Die Krystalle treten ofters in ilen Oelzellen des Holzparenchyms auf. 
wiihrend in demselben Schnitte die Oelzellen der Markstralilcn unJ des 
Libriforms mit gelbem oder firblosem Oc! erfiillt sind. 

Dicse Krystalle sind firblos, wcich und ist (.lie Krystallform sehr 
undeutlich. Viele dieser Krystalle sind v.u .\ggrcgaten vereinigt. Fig 
29, a. 1). Dieselben loscn sich sehr leicht in Alkohol oder Aether ohne 
Riickstand. Durch Erwarmen verfliichtigen sie sich vollstandisj-. 



386 Homi SIiiras:nva : 

Die bei dcr Untcrsuchung- ani^cwendeten Mcthodcn uiid Rcaktionen 
sind folgendc : 

I . Kochcii. 

Kleinc Holzspanc wurdcn in cincm Bccher zwci Stundcn lang niit 
Wasser stark gekocht. Das farblose Ocl uad die Krystallc warcn vollstandig- 
vorfliichtigt, aber das gclbe Oel fast unverandcrt. Bei der Untersucliung 
von im eisernen Kessel mit Wasserdampf destillierten Holzspiinen, habe 
ich diesclbe Erfahrup.g gemacht. 

2. ILrJiitzcii. 

Die Schnitte \vurden auf dem Objecttrager einige Alinuten lang hingsam 
erhitzt. Hierbei verfliichtigte sich das Oel und die Krystallmasse giinzlich, 
wahrend der in Alkoliol unl()sliche niit Jodgriin und l^^uchsin filrbbare 
sclnvammige Klumpen zuriickblieb. * 

3. ^ubliuiatioii 

I'lrliitzt man kleine Schnitte des Ilolzes im Sublimierglaschen schwacli 
und langsam einige Stunden lang, so sublimicrt die krystallinisclie Substanz 
(Kampher) und bildct feine Krystalle auf der kalteren Flaclie des Gliischens ; 
zugleicli biltleii sicli dort farbk:)se Oeltroi:)ren, wabrend das gelbe Oel fast 
unvi'randert in der Zelle zuriickbleibt. 

Anstatt des Sublimierglaschens lassen sich auch ausgehohlte Object- 
triiger verwenden, die ll()hluni;- muss mit einem Deckgliischen bedeckt 
werden, welches sorgfi'iltig anzutlichten ist. Der Kamplier krystallisiert auf 
der Innenseite iles Deckglases, wo sich auch das lli'ichtige Oel niederschlagt. 

4. /•'(irhiinx'Si'finx'i'iifii'ii- 

a. Alkainiatinktur. 
1 )ie in tier sehr \crdiiiit>ii fast alkoholfreien L()sung i'llier 10 Stimden 



I 



UelKT Kiilstcliiiiiir 1111(1 V<'itli(iliiiiff dcs Kaiiiplicrs iiii KaiiiplHMljauiiic. 3*^7 

lang liegcnclen Sclinittc zcii^cn das t^clbc Ocl intcnsiv rot ,L,^cfarbt. Die 
Krystalle und das farblosc Ocl sind durch das lant^'c Lict^an in dcr Tinktur 
niclit mehr sichtbar. 

b. Osmiumsaurc. 

Die 1% LosLing dcrselben briiunt des gelbe Oel, dagegen wirkt sie 
kaum auf das farblosc Ocl cin. Jedenfalls ist die Einwirkung auf das in den 
jiingcrcn Organcn cnthaltcnc Ocl starker als auf dasjenige des altcn Holzcs. 

5. Lusunginittcl. 

Die Loslichkeit des im altcn llolz vorkommenden Ocls ist selir ver- 
schicdcn. Im folgendcn ist das Vcrhaltcn des Oels gegeniiber den einzelncn 
Losungsmitteln wicdcrgcgcbcn (Die L(')sliclikcit des I fandclskamphers ist 
bci jedcm Losungsniittcl angcgeben). 

a. Alkoliol ir)st : 

das gelbe Oel, wobci cine farblosc schwamniigc odcr nctzartige ^lasse 
zuriick blcibt ; das farblosc Ocl Icicht, olmc Ri'ickstand ; die Krystalle 
Icicht, ohnc Kiickstand ; den I landclskaniphcr Icicht, ohiic Riickstand. 

b. Aether lost : 

das gclbe Ocl Icicht, niit Riickstand ; das f irblose Oel schr Icicht. <>hne 
Riickstand; die Krystalle schr Icicht, ohnc Riickstand; k\k:\\ Ilandels- 
kamphcr schr Icicht, ohnc Riickstand. 

c. Chloroform lost : 

das gclbc Ocl Icicht, mit Riickstand ; tlas farblosc (~)cl sclir Icicht, ohnc 
Riickstand; the Krystalle schr Icicht. ohnc Riickstaiul ; (V\\ Hamiels- 
kamphcr schr Icicht. (')hnc Riickstand. 

d. .\ceton hist : 

das gclbe Oel, mit Riickstand; das farblosc (^cl leicht. ohnc Riickstand; 
tlic Krystalle Icicht, ohnc Riickstand; (\k:\\ I l.uulclskampher leiclit. olmc 
Riickstand. 

e.Rcnzol Idst : 
das gclbc (^cl nach langcrem l-!in\\ irkcn, mit Riickst.md ; das larblosc (~)el. 



3^^'^ Homi Shirasawa : 

oliiic Riickstand ; die Krystalle leicht, ohiic Riickstancl ; den Handcls- 
kamplier leicht, oline Riickstand. 

f. Petrokacther l()st : 
das t^elbe Ocl scliwer mit Riickstand ; das farblose Oel leichter, mit Riick- 
stand ; die Krystalle. ohne Riickstand ; den Handelskampher, ohne Riick- 
stand. 

i^. Eisessig lost : 
das gelbe Oel erst nach der einiy;en Minuten, mit Riickstand ; das farblose 
Oel leicht, ohne Riickstand, die Kr}'Stalle leicht, ohne Riickstand ; den 
Handelskampher leicht, ohne Riickstand. 

h. Chloralhydrat iSo%) lost : 
das gelbe Oel langsam, mit Riickstand, das farblose Oel ohne Riickstand ; 
die Kr)'stalle leicht, ohne i^iickstantl, ilcn Handelskampher leicht, ohne 
Riickstand. 

6. Vcrlialtcii !:;ci:;c)i Saiiroi iind Alkalioi. 

a. In cone. Schwefelsaure ist : 

das gelbe Oel theilweise loslich und bleibt cine schwammige Masse zuriick ; 
das farblose Oel loslich, ohne Riickstand, die Krystalle lr)slich. ohne 
Riickstand ; der Handelskampher ]()slich, ohne Riickstand. 

b. In Salzsiiure ist : 

das gelbe Oel fast unl<')slich, das farblose Oel schwer oder kaum hislich, tlie 
Krystalle kaum l(")slicli ; (\<cw Handelskampher kaum h'xslich. 

c. In Salpetersliure ist: 

das gelbe: Oel theilweise loslich und (luillt die Oelmasse auf, infolgedessen 
sic oft aus der Zelle hervortritt ; das farblose Oel leicht li'islich, ohne Riick- 
.stand ; die Krystalle leicht loslich, ohne Riickstand; der Handels-Kampher 
leicht ]()slich ohne- Riickstand. 

d. in Kalilaiige ist : 

lias gelbe Oel unloslich, jedcch tritl Hr.iimiing desselbeii ein ; das farblose 
Oel unl.islich, ohne Fiirljung ; die Krystalle kaum l()slich. ohne Farbung ; 



I'cber Eiitslclmii^ iiikI Vertlieiliiiiy: dvs Kaiiiplicrs iiii KaDiplierbaiiiiic 3^9 

dcr Ilanderskamphcr kaum loslich. olinc FilrbuiiL^. 

1\6 fragt sich nun, in wclchcr l^ijzichung dicsc \'crschicdcnc Oelinhalte 
zu cinandcr stehen. Ks schcint mir, dass dieselbcn li()clist wahrsclicinlich 
Uebergangsstadicn zu cinander darstellen. 

Tritt Luft zum gclbcn Ocl liinzu, so bildcn sich in dcmsclbcn zalilrcichc, 
fcinc Blascn, wclchc ilini cin luidurchsiclitigcs, triibcs Anschn x'crlcihcn 
Fig. 20. Dicsc Erscheinung ist vicllcicht aufcinc Oxidation zuriickzufiihrcn. 

Bei liingerer Luftcinwirkung vcrgr()sscrn sicli die l^U'ischen, fliesscn 
zusammcn und bildcn grossc BLascn cntwcdcr in dcr Mittc dcr Zellc odcr 
an dcr Zclhvand, Fig. 21. \'on diescm Zcitpunktc an wird das Oel 
hellgclb und ctwas durchsichtigcr. Ini wcitcrcn Wndaufc wird cs immer 
klarcr und gcht in cine farblose fliichtigc, in Alkoliol Icicht loslichc 
Fliissigkcit i'lbcr, d. h. cs wird zu farbloscm Ocl I-'ig. 22-27. ^^^ ^'^^ diescm 
Prozesse die Menge dcs Ocls in dcr ZclIc abnimnit, odcr ob erst nacli dcr 
Uniwandelung in fluchtiges, farbloscs Ocl cine Abnclinien dcsselben zu 
verzciclincn ist, kann ich niclit bcstimmt sagen. 

Die Krystallc bildcn sich in diesen Ocl, und unterlicgt cs niir kcincn 
Zwcifel, dass dicsc Krystallc ^yKauiphcr'' sind. Fig. 28-29. 

Die Oclzcllcn, welclie fvir die friihe Krystallbiidung in Bctracht 
konimcn, liegen, wic ich konstatiercn konntc, ini Parenchymgewcbc. Das 
Vorkommen dasclbst liisst sich dadurch crklaren, dass im Parenchym dcr 
Luftzutritt wegcn seines lockercn und diinnwandigcn (icwcbcs schr cricichtcrt 
ist. 

III. Vertheilung. 

Kiios/'cii uiid Blatter. 

\\ ie ich Iriihcr gczeigt habe. sinil die Oclzcllcn in den tViihcstcn 
Statlicn tier l^ntw ickelung, unniittclbar hintcr dcni \"cgatationspunktc 
und auch in den iinursten l^lattanlagcn angclcgt wordcn ; derglcichcn auch 
bercits die Schleinmicnibriin und tiic rcsinogcne Schicht. 

Dicsc Ihcilc tier Ptlanzcnorgane bestchcn vollstandig aus mcristcnia- 
tischcni Cicwebc, welches thirch cone. Schwcfclsaurc vollie" zersttirt wird. 



390 



Hoini Shirasawa; 



Darin finden sich abcr bedeutcnd t(rosscrc rundlichc Zellen. Sic sind in 
der Xiihe dcs Vegetationspunktcs im mittlcren, bci dcr Knospcnachsc im 
ausseren Theile zu finden, Fig. i. 

Bei den Knospen nimmt die ZcUenzahl mit der Grosse der Blattan- 
lagen zu ; also erhalten die ilusseren Schuppen eine grossere Anzahl Zellen 
als die innern. Auf eineni Ouerschnitt durch die ]\Iitte der Knospe babe ich 
in einigen Fallen in iler fiinften Klattanlage 5 bis 10 Zellen gefunden, 
wiihrend sich in der zweiten auf dcmselben Schnitt nur 2 Zellen vorfanden. 

Bei dcm weiter nach aussen liegenden Blattanlagen, in wclchen schon 
die rudimentaren Gefassbiindel entstanden waren, niachtc sich eine sehr 
bedeutcndc Zahl von Oelzellen bemerkbar, und letztere waren in den 
inncren und ausseren oder audi in den niittleren Partien der Blattanlagen 
vertheilt. 

Die mittlcren oder die sich densclben nach aussen anschliessenden 
Telle sind jcdoch reicher mit Oelzellen versehen. 

Bei den Deckschuppen, welche durch die dickwandigen und aufihrem 
Ouerschnitt vicrkantig erscheinenden Sclereiden und auch durch die 
lange Behaarung auf der Aussenseite — besonders am Rande derselben — 
charaktcrisiert werdun, sind die Oelzellen mcistens im ilusseren Theile 
angclegt. Die Sclereiden der inneren Schuppen v\eiscn, in der Regel, noch 
eine diinne W'andung auf, und sind in ihrem Auftreten sehr sparlich. In den 
ausseren Schuppen ist ihre Zellwandung jedoch stark verdickt untl liat 
manchmal gabelf()rmige Kanide. Solchc Sclereiden, liegen besonders in 
i\cn inneren Randpartien iler Deckschuppen zahlreich beieinander. 

Bei jiingeren Blattern, welche sich bis circa 2,5 cm. Lange entfaltet 
haben, sind die (Jelzcllen uimiittelbar unter der l''.pidermis zwischen dem 
Palissadengewebe gclegen, zeichnen sich durch eine eigenartige Form, 
analog deu I'alissadciizillen aus, sind aber innner breiter als die letzteren 
und besitzen eine dicki:ri: Zellwand. Die, welcl'e zwischen dem Schwannn- 
parenchym auftreten, sind meistens kreisrund otler i llipsoidisch uiul etwas 
kantig. 

Bei (kn xoilkonuuen mtwickelten F)Iattern zeigen die (Oelzellen eine 
ganz ausge])ragte (iestalt, und zwar erscheinen sic zwischen dem Palissaden- 
gewebe ovalrund, im Schwammi)arcnchym runtllich oder cllipsoitlisch mit 



Uober Eutstcliiing uiul Verthoiliing' (Its Kamphcrs iiu Kaiii]>h!'rl):iiiiiii' 



591 



grosser Achsc parallel dcr Blattflachc. Audi sincl sic nicht mchr kantif,^ 
soiidcrn xollit^^ abgcrundct. Dicsc Formilndcrung verdankt vicllciclit ihrc 
Ursprung" dcr Spaniuing dcr Zellc. wclchc durch die Oelbildung in dcm 
Zellraum verursacht wurde. So schcn wir. nicht selteii, die Palissadenzcllen 
oder die Schwammparenchymzellcn durch die gross entwickelte Oelzellc an 
den Xachbar gcdriickt. 

Vom morphologischcii Stand punktc dcr Ausgcstaltung dcr Oelzellcn 
aus kann man sagen, dass ihrc Gcstaltung imnicr mit den umgcbenden Gcwe- 
ben in Zusammcnhang steht, z. B. die Oelzellen sind alle in ihrcn friihesten 
Entwickclungsstadicn bci den jiingsten Blattanlagen und Knospenschuppcn 
und dcm Vegetationspunktc cts. fast ausnahmslos rundlich ; bei den bereits 
entwickcltcn Pflanzcnorgancn, je nach dcr Stellc ihrcs \'orhandenscins. 
wciscn sic dagcgcn cine diffcrcnte Gestalt auf, bchaltcn abcr noch die 
charaktcrlstische Gcstaltung ihrer Abstammungszelle in dcm entsprcclun- 
den Gewcbe. 

Was die Zahl, Wrthcilung und Anlage dcr Oclzellen in der Blattspreite 
anbctriftt, so habe ich vom Rande ausgehend, bei vollig entwickcltcn cin- 
jiihrigen ]31attcrn das auf folgendcr Tabelle dargestellte gcfunden. 



l.angc 

das 

5-clinittes 

m.m. 


I )ic Zahl dcr 
Oclzellen 
zwischcn 

dem 
Palisadcn- 
gcwebe. 


Die Zahl dcr 

Oelzellen 

unmittelbar 

den Palissa - 

den gewcbe 

anschliesscnd. 


Die Zahl der 
Oelzellen 
zwischcn 

dem 
Sch.wamms- 
parcnciiym. 


Die Zahl der 
Oelzellen 

unmittelbar 
h inter der 
Epidermis 
der Blatt- 
unterseitc. 


Die Zahl dcr 

Oelzellen 

am 

Randgewcbe 
anliegcnd. 


c' 


Durchsch- 

nittlichc 

Zahl dcr 

Oelzellen auf 

mini liinge 

des Schnittcs. 


5 


8 


1 

J 


8 


I 


I 


20 


4 


6 


,0. 


5] 


"1 






— 

-/ 


4-5 




7) 


7 


9 








3»^ 


7 


6 


8 


,9 




T 




3-'^ 


8 


1 1] 


7 




151 








4.1 




6). 


1 1 




15 








4. 




i8| 


3 


19 








4- 


4.6 


9 










I 


I 








12 


10 


17 








39 


4-3 



392 



Homi Shirasawa : 



Die vorliegcndc Tabcllc zcigt, class das Auftrctcn dor Oelzellcn 
zwischcn odcr direct beim Palissadenij^ewebe haufig-er ist als bei den 
anderen Geweben, und dass auch auf dcr IMattunterseite. unmittelbar unter 
der Epidermis oder zwischen den l^pidermiszellen sich nur ausnahmsweise 
Oelzellen finden. 

Bei t\cn Blattstielen liei^t das Gefilssbundel in der ?vlitte und die Bast- 
zellen umschliessen den Sieb- und Holztheil. Die Sclereiden sind rings um 
das Gcfassbiindel unregelmassig verstreut. Sie sind bald zahlreich bald 
spiirlich vorhanden. Ich babe bei deni trockenen ^Material aus Java ein 
hiiufigers Vorhandensein derselben bcnierkt. als bei deni frischen ^Material 
vom botanischen Garten Bern. In letzten Falle war ihre Zahl unbedeutend. 
Die Oelzellen finden sich peripherich zwischen den Subepidermalzellen 
oder unmittelbar hinter denselben. Sonst sind sie zwischen dem Rinden- 
parenchym vertheilt. Auf dem Langsschnitt sind sie oft in verticalen 
Reihen, aber niemals in (iruppen x'ereinigt. Retzteres ist besonders bei 
di^n Randpartien leicht bemerkbar, Fig. 30. 

Im Gtfassbiintlel zeigen sie sich zwischen dem Parench}'m des Sieb- 
theiles, in 1 lolztheil dagegen nicht. Sie sind ziemlich klein mid erscheinen 
auf den Langsschnitt c^dindrisch (xler liinglich oval. 

Diejenigen Oelzellen, wclchc loci den Subepidermalzellen angelegt 
worden sind. zeichnen sich in der Kegel, durch ihre (irr)ssc gegenueber 
ilcn XachbarzclKn aus, widtrend die im Riiulenparencln'm ihnen an Grosse 
fast gleich kommen. Auf dem (Juerschnitt erscheinen sie meistens rundlich ; 
auf den Lilngsschnitt weisen sie dagegen eine oxale oder elliptische Gestalt 
auf. 

Die Oelzellen der Jilattstiele zeichnen sich oft durch ihre L'mgebung 
aus; die Pareiichymzellen sind niimlich rings um die ( )elzellen strahlen- 
fi'irmig angeordnet, so tl.iss die ( )elzellen in der .Mitte der Zellgruppe sitzen. 
Dieses kann man .lus den Ouer- oder Rangcschnitten ersehen. h'ig. 31 a. b. 
In Hvzug auf die Zahl der ()elzellen der Blattstieles ergab eine I'nter- 
suclumg \'ermittelst diiimer nuerschnitte bei eiujidirigen frischen HUittern 
folgendes : 

Im untcnii und nu'tti.ren leile des Blattstieles enthalten die peripiieri- 
sclun Reihen 23-30 (Oelzellen, die der Hlattsjireite sich anschliessendeii 



Leber Eiitsteliiiiig uiul Vertheilniig: lies Kainphcis iiii Kaiiiplieihaiiiiu'. 393 

Thcilc nur 15-23. Hcim trockcncn ^Material aus Java odcr aus Japan zilhltc 
ich tlai^ct^cn 29-35. was mit davon iibcrzcu^te, class das in einhcimisclicn 
Gegcnden L^cwachscnc Kxcmj)lar nichr Oelzellen zu crzcuj^cn vcrmai^, als 
dasjeni^^c ini Treibhaus dcs botanischcn Gartens. Dicsc Tatsachc bewcist 
uns, dass die klimatische, und Standortsxxrhultnisse zur Oelzellbilduni,^ in 
Keziehunti; stehen, was Tschirch' schon bemerkt liat. 

Die Parenchymzellen des HIattstielcs weisen Tiipfel auf. Bc'i (km Oel- 
zellen fehlen abcr diese Tiipfel vollstandii;". wenngleich man nicht selten 
die Tiipfel der Parenchymzellen bemerkt. welche bis an die Mitte der ge- 
meinsamen Wandung heranreichen. 

Die derbe Wandung, das kY-hlen der Tiipfel. und die Korkbildung in 
der Zelhvand bilden tlie charakteristischen l^Ligenschaften der Oelzellen. 
Vielleicht wird damit der Wecliselv'erkehr mit den Xachbarzellen erschwert 
und tlie Ablagerung untl Zuri'ickhaltung des Oels am Orte der Krzeugung 
m<")L'lich uemacht. 



K/uc/c 



Die Oberhaut der Rinde des einjahrigen Triebes ist stark cuticularisiert. 
Sie farbt sich mit Alkannatinktiu' intensiv. Beim Abwaschen mit .\lkohol 
und W'asser halt die l^^iirbung ziemlich lange Zeit an. Die Rindenparenchym- 
zellen sind meistens tangential gestreckt und ziemlich dickwandig. 
Zwischen ilcn I'arenchymzellen timlen sich sehr zahlreiche Schlcimzellen. 
Die (Oelzellen sintl noch spilrlich entwickelt. Sie sind meistens peripherisch 
an die ICpiilermalzellen ange(M-tlnet. und auf dem Ouerschnitt runtllicli. auf 
dem Liingsschnitte li'inglich (»\al oder oft rundlich und bedeutend grcisser 
als die benachbarten Zellen. l-'ig. 32, 33. 

Ini Siebtheil des Ciefiissbiindels kommen sie selten \"(ir. im 1 lolztheil 
dagegen noch gar lu'cht. .\nschliessentl an die aussere Randpartie tier 
Bast/,ellgrup[K' treten oft Ix'deutentl grosse Oelzellen auf 

Bei tier l\intle ties 5 jilhrigen Trielies ist ties Teritlerm noch nicht 

^ 'I'scli'rcli. AiiatomisclKT Atla< S. 132 u:-.<l llaiv iind I laivheluiltor. S. 3S9. 



i 



394 



Homi Shirasawa : 



aufgctrctcn. Die l^pidcrmis ist sehr stark vcrdickt. besondcrs beim Material 
aus Ja\a, uiid rai^t auf dcr Innenseite buckclic^- licrvor. 

Die Schleimzcllcn siiid hicr schr reichlich cntwickclt. Die Oelzcllen 
kommen sowolil in der primilrtn als auch in der sccundaren Rindc vor. In 
der letzteren treten sie, in der Regel, liaufiL^er als in der crsteren auf. 
Diejeni<^fen, welclie in der priniiiren Rinde vorkommen, sind etwas 
abi;eplattet, und in tanq-entialer Richtung lang" gestreckt, wilhrend sie in der 
sekimdiiren Rinde eine rundliche odcr liing^lich ovale Form zeigen. 

Auf der alten Rinde bildet sich cine starke Borke. In der sekundiiren 
Rinde kommen stark verdickte. in radialer Richtung gestreckte Scler- 
eiden \or. Sie vereinigen sich meistens reihenweise in tangentialer Richtung. 

Die Oelzcllen mit eincm gclben Inhalt, sowohl. als auch Schleimzcllcn 
treten hciufig in sckundarer Rindc, sehr scltcn in primilrcr auf 



Holr. 



J)ei der L'ntersuchung ties frischen Matcrialcs habe ich nicmals das 
\'orkommcn von Del in (X'cw Zellen das Ilolzthciles dcr cinjahrigen Tricbc 
bcobachtct, wcnngleich im ]\Iark, besondcrs in (\^w iiuscrc n Thcilcn dcssclbcn 
zicmlich \iclc Oelzcllen vorhanden warcn, h'ig. 34. 

Im Mark treten oft sehr verdickte Sclercidcn auf Sie kommen x'crein- 
zclt (iLJcr zu Xcstcrn vcrcinigt vor. 

]-5cim zwcijilhrigen Tricbc aus Jaxa treten die Oelzcllen im crstcn 
Jahresringe auf willircnd im zwcitcn Jahrcsringe ihr W^rkonnnen noch 
kaum bcmerkbar ist. 

Im 'I "ricbe (ks (h'ittcn Jaluxs habe icli dicsclbc k'.rfduamg gcmacht ; 
in crstcn und zwcitcn Jahrcsringe konuncn schon zahh-cichc Oelzcllen vor, 
dagcgcn im drittcii d. li. im ji'ingstt-n jahrcsringe sind noch keinc Oelzcllen 
\ orhanden. 

Im Mark tritt das ICntgegcngcsctztc cin, iiulcm in dem cinjilhrigcn 
rricijcn iWv ( )clzcllcn vcrhiiltnissmiissig zahh'cichcr als in den zwcijuhrigen 
Tricbcii imd in Ictztcrcn zahhxichir als in dreijrihrigcn auftrcten, sodass mit 
ilcm .\Itcr dcr Tricbe' die Anzahl '.Vx ( )il/,rlKii abiiimml. 



I'eber Entstehung' imd VcrtlK iliiiig' dcs Kami>li<'rs iiii Kaiiii»li<'rbaiiiii<'. 395 

Was clas Vorkommcn unci die Wrthciluuf;" dcr Oclzellcn in Holz 
anbctrifft, so babe icli hauptsachlich clas altc Holz zur Untersuchung bc- 
nutzt ; da in dicscm das Gewcbe den ICndpunkt seiner Ausbildunc^ errcicht 
hat, unci daher fiir die Untersuchung' ein sehr bec|ucmes unci klares Object 
abg-ibt. 

Die Oelzellen treten im Holz zwischen den primilren und sekundiiren 
Markstrahlcn, im Holzparenchym und im Libriform auf. 

Sie finclen sich ausschliesslich am Rande cler Alarkstrahlen, niemals in 
der Mitte clcrselben. Sie sincl meistens ket^elformig mit der in clas aussen 
liegende Gewebe eindrini^enden Spitze, und bedeutend grcisser als die 
iibrigen Zellen der Markstrahlcn Fig. 35, 36, 37, 40. 

Diejenigen, welche zwischen das Libriform eingebettet sincl, sind liing- 
lich oval und etwas zugespitzt. Jhre Zellwand ist ziemlich \'erdickt, wie die 
der Libriformfasern Fig. 38, 39. Die in dem Parenchym liegenden Oelzellen 
haben die grosste Ausdchnung. Sie sind elliptisch un.d finden sich manch- 
mal mehrere neben einander liegend Fig. 41, 42. 

Bei cliesem dreifachen X^orkommen cler Oelzellen treten die im Paren- 
chym liegenden in den \^ordergrund, da dieselben sowohl an Zahl als auch 
an Grosse die andern iibertreffen. 

In einetii Jahresringe enthiUt das Iderbstholz meh.r Oelzellen als das 
Friihjahrsholz. Bei nieiner Untersuchung der Jahresringe habe ich auch 
bcmerkt, dass von einer bestimmten Grcnze an im Friihjahrsholz zum 
Flerbstholz him die Oelzellen zahlreicher auftreten. Diese Tatsache erkliirt 
sich dadurch, dass fiir die Oelbildung gewisse l^'aktoren \'orhanden sein 
miissen. wobei wahrscheinlich die ]'".rh()hung der Temperatur keine geringe 
Rolle spielt. 



jr///-.:.r/. 

Bei cler W'urzelspitze, welche ausschliesslich aus weichem meriste- 
matischen Gewebe besteht, fand ich einige Zellen, welche durch ihre Form 
von den uebrigen Zellen ausgezeichnet sind. Der Tnhalt derselben reagierte 
weder mit Alkannatinktur noch mit Osmiumsi'iure. 



596 



Hoiiii 8]iirasa>va ; 



Bci dcr ctuas cntwickclten W'urzcl warcn die Oclzcllcn schon bcmcrk- 
bar. Sic sind ini Rindeiiparcnchym nebcncinai'idcr angclct^t worden. Der 
Oclgchalt in ihncii ist iK^ch i^crini^. In dcr cinjahrit^cn Wurzcl ist das 
Vorkomnicn dcr Oclzcllcn bcdcutend, wcnn^lcich sic hauptsiichlich im 
Rindcnparcncln'ni abcr nicht im Holztheilc dcs Gcfiissbundcls crscheincn. 
Der Oclinhalt ist gclb luid bcdcutcndcr als in dcr jiinocrcn Wurzcl. 

Bci dcr altcn Wurzcl ist das \'crhaltcn dcr (V'lzcllcn ahnlich \vic bci 
dcm altcn Holz. 



IV, Zusammenlassiing der Resultate. 

Fasst man die vorstchcndcn Untcrsuchunc^cn zusammcn, so lasscn sich 
folLfcndc Schliissc zichcn : 



A. Ucbcr die li)itstL'huug. 

1. ]^ci Cinnamomum C'amphora cntstchcn tiic Oclzcllcn schon friih 
ummittt Ibar hintcr dcm \'cij;ctationpunktc. 

2. J^ci juni;crcn Ptkinzcnori4ancn ist cU-r Inhalt tier Oclzcllc ,.Acthcri- 
schcs Ocl". 

3. ]-)icscs Ocl bildct sicli in dcr \on Tschircli hcn.mntcn ,,rcsino_<;cncn 
Scliicht", wic bci tlcn andcrcn Laurinccn-l'lkinzcn und dicsc rcsinoi^cnc 
Mas!:;c blcibt schr lan<^c Zcit in dcr Zcllc crhaltcn. 

4. In den jiinL,a'ren rdanzcnornancn durchtriinkt das Ocl die rcsin(^_L^cnc 
Masse, im tropfenformigen Zustandc kommt cs schr seltcn vor. 

5. l>ei k\v\- in troj)ischcn Gcgcndcn (Jawa) j4c\\achscncn rilanzcn hat 
tlas Ocl rcsp. die resinocj^enc Masse cine dichterc- C'onsistenz. und die Menge 
dessclben ist _L;rr)sser als bci tlcn im Treibhaus (im botanischen (iarten l^ern 
und Miinchen) t;cziichtetcn Ivxemplarcn. 



robor Eutstehnuj;- iind Yortheiluiig des Ka!ii])liPis iiu Kainplierlmiiiiic. 397 

6. In l^lilttorn kommt das Sckrct oft in b(jutclf<)rmi<^cn Hilutclicn vor. 
(bci clcr Untersuchung von friscliem Material). 

7. Bei den alteren I-51attcrn tritt das Oel reiclilicher als in jungcrcn 
]31attern auf. 

8. Im alten Holz nimmt das Oel eine orangegelbe Filrbung an ; dieses 
Oel geht spiiter (durch Sauerstoffaufnahme ?) in das farblose Oel fiber. Aus 
dieseni bildet sich der krystallinische Kamphcr. 

9. Dieser Umwandlungsprozcss geht erst nach einigen Jahren vor 
sich ; jedenfalls erst lange nach deni Abschluss der Oelbildung in dcr Ocl- 
zelle. So ist im alten Holz die Relativmenge von farblosem Oel und der 
Krystalle bedeutend grosser als die von gelbem Oel ; dagegen im jungen 
Holz iibertrifft die Menge des letztcren die erstere. 

10. Die Oelzellen, welche zwischcn dem Tarenchym liegen, enthalten 
mehr farbloses Oel und Krystalle als die in anderen (jeweben. 

11. Wenn sich bei alten Sti'immen Kamphermasscn in 1 lohlungen und 
Spalten des Holzes finden, so konnen sie dorthin nur aus den Oelzellen 
durch Sublimation gclangt sein. Sie befinden sich also an ,,Secundarer 
Lagerstatte". 

12. Durch die jetzt iibliche Methode der Kamphergewinnung ist es 
kaum moglich, das gelbe Oel aus dem Holz zu erhalten, wenngieich das 
farblose Oel und die Krystalle leicht destilliert werden konnen. 

Ik [\'ln'r die ]^crtJu'ilitng. 

1. ] )ie Oelzellen kommen vor: 

a. bei den jiingsten Blattanlagen in der .Mitte oder den derselben 
anliegenden Theilen ; 

b. bei (\c\\ De'ck'schuppen tier Knospen \"or\\iegend in ^Xcw iiusseren 
Partien. 

2. Hei \-()llig entw ickelten Hlilttern fuidet man sie im Talissaden- und 
Sch\vannnparench\-m. .\m Schlusse ihrer h'ntw ickelung besitzen sie eine 
abgerunelete l"(M'm. 

3. Bei ([^-w in tropischen ("regemlen gewachsenen l-'xemjilaren sind die 



598 



Hoiui ShiiMsawa. 



Oelzcllen zahlrL'ichcr als bci den im Treibhaus gczogcncn. Dieser Unter- 
schied fiillt hauptsaclilich bei den Blattstielcn ins Augc, wahrcnd bci dcr 
Blattspreite dersclbe wcniger wahrnehmbar ist. 

4. In den Blattstielcn sind die Oelzcllen verhaltnissmassig zahlrcichcr 
als in den anderen Organen. 

5. Im liolzthcile des jungen Gefiissbiindels iind zwischen den Epi- 
dermiszellen babe ich nienials Oelzcllen bemerkcn konnen. 

6. In der secundiircn Rinde befinden sich mehr Oelzcllen als in der 
primiiren Rinde. 

7. Im Blattsticl und im der Rin.de des jiingsten Tricbes sind zahlrcichc 
Schleimzcllen vorhanden. 

S. Im jungen Mark sintl die ». )c]zcllen schr zahlreich. mit dcm Alter 
dcsselben nehmen sic ab. 

9. Im 1 lolz des einjalirigen Tricbes sind die Oelzcllen kaum vor- 
handen, wennglcich im z\veijahrig"en Holz ihr Auftrcten schr bedentcnd ist. 

Sic befinden sich zwischen den Markstrahlzellen, dcm Holzparenchym 
und dcm Libriform. 

In den IVIarkstrahlcn treten sic ausschlieslich an dcm Rande auf. Das 
Holzparenchym cnthiilt in der Regel mehr Oelzcllen als die anderen 
Gewebe. 

10. Im llcrbsthojz des Jahresringcs sind mehr ( )elzcllen als im 
I'riihjahrsholz. 

11. In tier W'urzel kommen die Oelzcllen auch \or, und ihre X'ertcil- 
Liiig und ICntwick-chmgcschiehtc ist diesclbc wic bci den (^berirdischcn 
Teilcn der I'fianzc. 



Figuren-Erklarung. 



Abkiirzungcn : 
I'"p. = I'lpidermis. 
foe. =:farbloses ( )cl. 
gf. = (iefiiss. 
kr. - Kr\-stall. 



C = Caml)iuni. 
]'".pz. = I'.piikrmiszcllc 
goc. — gclbis ( )el. 
lip. = I lolzpari. iicluni. 
L. = I aimcu. 



L'eler Eiitstehniia: iiinl YcrtlH'iliiii^ tics Kamiilicrs iiii Kaniplicrbciimc. 399 

lb. — Luftblascn. lib. = Libriform. 

ms. — - Markstrahlcn. Oe. == Oel. 

Oez. — Oelzellc. p.— I'lasma. 

reg. = resinogenc Scliicht. 

Schl. = Schlcim, Sclcr. — Sclcrciden. 

Fig. 1. Liliigsschnitt ciiicr Knospc, uni die Vcrthcilung dcr Schlcim- 
rcsp. Oelzcllen und Sclcrcidcn in dcr Knospenaiilagc und Knospcnachse 
zu zcigen. 

Fig. 2. Erstc Stadicn dcr i^>nt\vickclung von Oclzcllcn : 

a. Die resinogenc Schicht tritt auf. 

b. BandfJh'mige Schlcimfadcn treten in der Mitte dcs Zellraunics 
zusanimcn. (Bcidc sind an Alkoholprilparatcn bcobachtef. 

Fig. 3. iM'stes Auftrcten von Sckret in der rcsinogenen Schicht 
Die resinogenc Schicht ist durch die Oelbildnng aufgeblascn. 

Fig. 4. Die resinogenc Schicht contrahicrt sich und bildct Lumen 
zwischen dcrsclben und dem Schleimbelege. (Die Prilparate sind aus der 
Knospcnachse). 

Fig. 5, 6. Dicsclben Stadicn der Oclbilchmg in (.]cn (3elzcllen aus 
eincni IMattstielc dcs cinjahrigen Blattes. 

h^ig. 7. Dieselbe aus cincm cinjahrigen Hlatt. 

Fig. 8, 9. Fast fertige Oclzcllcn aus der Knospcnachse. Die Reste 
dcr Schleinimembran sind 1:)emcrkbar. Die resinogenc Schich.t ist niit dcni 
Sckret durchtriinkt. 

1^'ig. 10. Oelzellc aus dcrsclben. Oel ist in cineni grossen bcutel- 
fth'niigcn Schlauchc vorhanden. 

Fig. 11-14, Oclzcllcn aus einjuhrigcn Hlilttcrn : 

k'ig. II, 12. Die resinogenc Schicht niit Oel ist contrahirt. 

Fig. 13, 14. Die durch Oelbildung aufgcblascne rcsin(\gcne Schicht. 
Die Reste der Schlcininicnibran sind noch \orhandcn und liic rcsinogenen 
Schlauchc in das niittlcrc Lumen her\ortretend. 

Fig. 15, 16. Oclzcllcn aus zwcijahrigcn I^lattcrn. 

h'ig. 17. Dicsclben mit zahlreiclun eifn'mig aufgcblascnen rcsinogenen 
Schlauchen. 



400 Hoini Shirasawa. 

Fig". 1 8. Oelzellen mit tropfenfonnigen Oelinhalt ; aus einem zwei- 
jiihrigen Blattstielc. 

Fig. 19. Oelzelle aus einem Blattstielc des trockenen INIatcriales aus 
Java. Das Oel ist im schaumigen Zustande vorhanden. 

Fig. 20-29. Oelzellen aus altem Holze, bei welcben die successiven 
Umwandlungsstadien von gelben Oel zu Krystallen zu sehen sind : 

Fig. 20. Das durch die zahlreichen feincn Luftbluschen triib ausse- 
hende gelbc Oel erfiillt den ganzen Zellraum. 

Fig. 21. Die kleinen Luftbliischen haben sich zu einer grossen Luft- 
blase in der Mitte des Oelinhaltes vereinigt. Das Oel hat ein klares 
Aussehen. 

Fig. 22. Erste Stadien der Umwandlung von gelben Oel zu farbloscn 
Oel. 

Fig. 23. 24, 25. Die weiteren Stadien desselbcn. Das farblose Oel ist 
anfangs schaumig (Fig. 24) und dann klarer (Fig. 25). 

Fig. 26. Im f^^rblosen Oel sind noch die Reste von gelbem Oel be- 
merkbar. 

Fig. 27. Ganz klares farbloses Oel. 
Fig. 28. In demselben bilden sich Kr}'stalle. 
Fig. 29. a. b. Krystallc aus dcm farblosen Oel. 

Fig. 30. Langsschnitt durch einen Randtheil voni l^lattstielc. Die 
Oelzellen sind in X'erticalreihcn angeordnet. 

Fig. 31, a. Oelzelle, welche von den I'arenchymzellen strahlenformig 
umgeben ist. (Aus dcm Ouerschnitte des Hlattstieles). 
b. Dersclbe aus dcm Liingschnitte desselbcn. 
hig. 32. Liingschnitt der Rinde des jiingsten Triebcs. Die Oelzellen 
siiul an die Fpidcrmis/.cllcn in Verticalreihen anlicgcnd. 
l""ig. 33. Oucrschnitt desselbcn. 

I'Mg. 34. Oucrschnitt cincs jiingstcii Triebcs. bei dcm die Vcrthcilung 
der Oelzellen bcnurkbar ist. (Mit .Alkannatinktur 1. 

l'"ig. 35. Oucrsclniitt des altcn llol/.cs. Die Oelzelle kommt zwischen 
i\cn Markstrahlzellcn vor. 

I' '.U- 3^'- Radialschnitt tlcssell)cn. Die Oelzellen licgen zwischen 
Randzcllcn der Markstrahlen. 



Ueber E!itst<'limig iiiul Vertheiliing- dcs Kampliors im Kaniplicrbaiime. 401 

Fig. 37. Tangcntialsclinitt dcsselben. 

Fig- 38. Ouerschnitt clcssclbcn, wohci die OlIzcHcii, wclchc zwischcn 
dcm Libriform lieg'en, sichtbar sind. 

Fig. 39. Radialschnitt desselbcn. 

Fig. 40. Tangcntialschnitt dcsselben. Die Randzellen der Mark- 
strahlen sind durch die Sekretbildung- stark vergrossert. 

Fig. 41. Oucrsclmitt desselbcn. Die Oelzcllen sind zwischcn dcm 
Parenchym entstanden. 

Fig. 42. Radialschnitt dcsselben. 




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BCLL. AGRIC. COLL. VOL. V. 



PLATE XXLIL 




Hp 



Investigations on Flacherie. 

PA' 

S. SaAvamura. 



Chapter I, Introduction. 

Flacherie is a frequent disease of the silk-worm, causing great damage 
to sericulturists. The first observation was made by Pasteur who regarded 
this malady as being caused b}' three species of bacilli and a micrococcus. 
These microorganisms penetrated into the tissues of the larva and even into 
the eggs. Cnboiii, however, was of the opinion that the pathogenic bac- 
terium of this disease was a micrococcus which produced black spots on 
mulberry-leaves. Recently IMacchiati ^ made investigations on this disease 
and concluded that the malady was caused by a streptococcus quite diffe- 
rent from that found on mulberry-leaves which was a diplococcus. He 
recognized this streptococcus as the Streptococcus boinbycis. He found 
besides this a bacillus in the digestive canal of the diseased larva, but 
he thought this bacillus could not cause the malady, its action consisting 
merely in the acceleration of the malady. 

According to ISIaceliiati the streptococcus and the bacillus have the 
following properties. 

Streptococcus bouibycis Ulacchiati. 

The cell is round or oval, the diameter being 1.25 — lo/i- It appears 
never isolated, but two, five or more unite in chains. It is aerobic ; the 
propagation on potato is \er}' quick aiul the coloin- thereon is greenish- 
yellow and has a metallic lustre. Colon\- on gelatine is light yellow, and 
gelatine is completely liquefied. 

1 Coiitribuzioiic alia Biologia dei Batten' lici I'achi atiocti da flaccidezza. Ix stazioni sperimcntali 
Aijraric Italiaiic. Vol. XX. T'avt. II. 



404 S. Sawamnra. 

Bacillus bombycis Macchiati. 

This bacillus exists not only in the larva, but also in the coccoon, 
crysalis and imago of silk-worm. The length is i — 3yx or often more ; the 
ends of the rod are round, and two or more unite forming a long thread. 
The 'cell-membrane consists of cellulose, as it is coloured blue by iodine 
and sulphuric acid. It is motile and produces spores usually in the mid- 
dle part of the rod. Colony on potato is yellowisli-brown and elevated, 
which turns afterwards light brown. 

Colony on gelatine is light yellow, and has a milky appearance, and 
gelatine is quickly liquefied. In gelatine stab-culture gelatine is completely 
liquefied, and flake-like precipitates are produced. 

After the investigations of Macchiati, Krassilschtscliik^ in Pasteur's 
Institute made some investigations on this malady, and found that it is 
caused by a micrococcus quite different from that of Macchiati. This 
micrococcus has the following properties. 

Streptococcus Pastoriaiuis Krassilschtschik. 

The cell is round, and lias a tlianieter of i — \.\fx. It occurs in the 
form of diplococcus. Colony on gelatine is round and gray, and gelatine 
is not liquefied. On gelatine stab-culture colony grows in nail-like form 
without liquefying it. 

Macchiati^ assumed that the micrococcus found by Krassilschtschik 
-was the same as his streptococcus, disregarding the great difference of the 
properties between his and Krassilschtschi/y s microbe. 

Macchiati-^ ])roposed to examine witli a microscojie the moth to be 
used for laying the eggs and to w.ish tlie eggs with sublimate solution, 
since the streptococcus exists usual I)- in tlie moth and eggs. lie said that 
very good results were obtained in \arious places by practising his 
liro]iosal. 

• C'omptes rcndus 1S9C. II. V. 427. 
' Socicfa botanica italiana 1896. 



Investigations on Flacherio. 4^5 

The microbes of flacheric were sometimes used for killing insects. In 
1890 Hoffmann'^ used various species of microorganisms to kill Laparis 
inonacJia, injurious insect of forests, and found that Botrytis and the microbe 
of flacherie could become parasitic on this insect and kill it. 

Tangl,- however, denied Hoffmaiui s report, since he did not believe 
Bacillus bovibycis was the true pathogenic microbe of flacherie, and he said 
that bacteria could not be used for killing Laparis, as there was not known 
any microbes that were joarasitic on that insect. 

Tubciif^ described a certain bacterium which became parasitic on 
insects of pine-trees and also caused flacherie. The infected insect lost 
appetite and died finally. The contents of the intestines were brown, and 
numerous bacilli were found in it especially in the fore-intestines. He gave 
this bacillus the name of Bacillus inonacJiac. Its width is o.5yLt and length 
i/i. It was present also in the blood of the dead larva. This disease 
broke out more frequently, when the climate was cold and moist. Tubcuf, 
henceforth, explained the reason, that the disease was produced, because 
when it was cold and moist, the food was not digested well and remained 
long in the intestines, thus offering an opportunity for the growth of the 
bacillus. 

In 1901 the .Vustrian -\gricultural Experimental Station* reported 
that flacherie could not be infected to healthy silk-worms, neither by 
giving together with mulberry-leaves the intestinal juice of the diseased 
larva, nor the pure-culture of the microbe. 

Oniori^ in Japan made investigations on this disease, and found that it 
was caused by four kinds of special micrococci and two kinds of special 
bacilli, and as the symptoms of the disease caused respectively by these 
microbes were different, he distinsjuished three kinds of flacherie. 



» Cenlral-151att fiJr Rikteriologic XI. 1*. 341. 

' „ .. ., XVI. r. 660. 

= „ ., .. XTI. r. 269. 

* OcsteiTcich. Wrsuchsstat'onen IV. I 'art 3. 
•'■• Xihoii Sai)l>vomii. 



4o6 S. Sawaniiira- 

Chapter II, 
Description of the Bacteria found in the Diseased Silk-worm. 

The diseased larva lose appetite, vomit a viscous fluid and suffer from 
diarrhea or excrete a viscous fluid in most cases. When they are dead, the 
third and fourth segments become somewhat elongated and the whole body 
softenes. But there rarely occurs a case in which the dead body shrinks 
and becomes rather hard. The dead worm turns black usually very soon.^ 
Sometimes while the diseased larvae are still living, the third and fourth 
segments arc coloured black. In the digestive canal there are found 
usually a viscous fluid containing very little fragments of mulberry-leaves, 
but sometimes the digestive canal is filled with the fragments of mulberry 
leaves compressed to a hard mass. The color of mulberry-leaves therein 
is usually brown, while that in healthy animals is green. But rarel)' the 
mulberry-leaves in the intestines retain the original green color. While the 
reaction of the intestinal juice of the healthy larva is strongly alkaline, that 
of the diseased is in most cases neutral or onl}- very faintl}- alkaline. ^ The 
fluid excreted shows often an acid reaction. 

In the intestinal juice there are foiuid a great number of bacteria. 
That abundantly found in many cases is a micrococcus. In some cases only 
micrococci are found, but in many cases there exist along with them large 
and short bacilli. There is sometimes a case in which short motile bacilli 
are seen, but it is ver)- rarely observed, except in silk-worms reared in sum- 
mer, that the large bacilh alone occur. Although these organisms exist 
abundant!}' in the intestinal juice of the diseased ]ar\ac, they can not easily 

» Acconliiit; lo Dc-itilz (.\icli. fiir Aiialoniic uikI I'hysiologic ig02. P. 32S) tlic turning black of 
the insect larva is due to oxydase?. 'I'lic fact that tlic diseased larvae turn usually very <iuickly black 
after death, or show black spots while still living, is probably due to the increased production of 
oxydizing enzyms accelerated by the insufilcicncy of nutrition as in the case of the vegetable cell 
which was proved by U'ooils (Central!), fiir Uaklcriologie IT. \'u\. 5. No. 22), or by the poisoning 
by nitrite. 

- The digestive enzyms of Lcpidopfeya arc active onl)' in an alkaline solution, and lose their 
action in an acid solution. S. Sn-vamura. This Bulletin vol. I\'. \o. 5. 



Investigations on Flaclierip. 407 

be detected in the tissues and blood of the diseased. It is clear from this 
fact that flacherie is caused by the propagation of the microbes in the 
intestinal juice. 

By investigating silk-worms reared in spring, summer and autumn, the 
writer found that the micrococci in the intestinal juice are not of a single 
species but of many. But as their form is the same they can not be distin- 
guished only by microscopical examination. The most remarkable diffe- 
rence is the color of the colonies on solid media. As in plate-cultures 
prepared from the intestinal juice of the diseased larvae, colonies of various 
colors made their appearance, it is beyond question that there exist in the 
intestinal juice of the diseased not only a single species but many species 
of micrococci (Fig. I and II). These microbes are present not onl}' in the 
diseased larvae, but also in the healthy ones, although the number is small. 
Their presence in the latter case can be detected with a microscope or 
more easily by preparing plate-cultures. 

To know whether the microbes are present in the interior of the eggs 
of silk-worm, they were washed with O.i^ sublimate solution and then 
with sterilized water, and crushed in bouillon, in which a micrococcus and 
a large bacillus propagated after few days. These experiments were 
repeated many times always with the same results. By examining the 
properties of the microbes, the large bacillus was found to be Bacillus 
inegathcriuin Dc Bary and the micrococcus a Sarcina, the properties of 
which will be described later on. The presence of microbes in the interior 
of the eggs of insects other than silk-worm was observed by Blockjiiaiiii, 
Korschclt, and Zaccharis. ^ 

The properties of the microbes found in the intestijial juice of the dis- 
eased larvae and in the eggs are as follows ; — 

The Large Bacillus. 

Form : The cell, when cultured in bouillon for 24 hours, is 0.8//. wide 
and 3 — 5^ long. In the intestinal juice it is larger. The extremities of 

* Caitral-RIatt fiir Bakteriologie II. p. 546 and XI. p. 234. 



40'^ *'. Smyamiira. 

the rod arc round ; and flagella grow on all sides, and are stained by 
Lofflcrs method. It exists isolated usually in the intestinal juice, but in 
nutritive fluid two or more are united. 

Spore-formation : Spores are easily formed usually in the middle part 
of the rod. 

Motility : It shows a slow oscillating motion. 

Gram" s method : Positive. 

Oxygen : The growth is better in presence of air. 

Bouillon : Propagation is good, and a cloud-like precijDitate is formed 
and a feeble ring on the wall of the tube. 

Gelatine streak : Gelatine is quickly liquefied along the inoculated line. 

Agar plate : Colony having curl-like appearance, irregularly extending 
from a light brown point formed in the centre. 

Agar streak : A dirty white colony is formed on the whole surface. 

Agar stab-culture : Colony is formed straightly along the inoculated 
line to the bottom, and it propagates on the surface quickly to the wall of 
the tube. 

Potato : An elevated gray colony within 2 days of inoculation at 20'C. 

Milk : Milk is coagulated but not with an acid reaction. 

Reduction : Nitrate is reduced to nitrite as shown by the iodine-starch 
and Gricss' reaction. 

Gas-Production : Gas is not evolved by cultivating in a nutritive solu- 
tion containing glucose. 

HoS : A trace of H-S is formed by cultivating it in bouillon. 

Acid-production : By cultivating for 3 days at room-temperature in a 
nutritive solution containing 5^ of glucose (with Ca CO3), there was pro- 
duced 0.i53^y of acid, calculated from the dissolved CaO, as lactic acid. 

By these properties this bacillus is proved to be Bacillus mcgatJicrium 
Dc Bary. 

The Sliort Bacillus. 

Form : I'hc cell cultiu'ed in bouillon for 34 hours is o.6/i wide and 
I.O — 1-5/14 long. It is isolated both in the intestinal juice ami in bouillon, 



Investigations on Flacherie. 4^9 

and very rarely two are united. The extremities of the rod are round. 
Flagella are colored by Lofflcr s method, and some have them in one end, 
while the other on all sides. 

Spore-formation : Spores are not formed. 

Mobility. It moves actively. 

Gram' s method : It is not colored by Grant s method. But some 
absorb colors especially well in the ends of the cell. 

Bouillon : Bouillon becomes turbid and viscous. The precipitate 
formed can be easily distributed by shaking. 
Gelatine plate : A white round colony is formd without liquefying gelatine. 

Gelatine stab-culture : Colony is formed straightly along the inocu- 
lated line to the bottom, and on the surface a white colony is formed which 
extends to the wall of the tube. The centre of the colony assumes a light 
yellow color after some days. 

Agar streak : A moist, bright, white colony is formed. 

Potato : An elevated yellow colony is formed. 

Milk : Milk is coagulated, acid reaction being produced. 

Gas-production : Gas is evolved by cultivating it in a nutritive solution 
containing glucose. 

Reduction : It reduces nitrate to nitrite. 

Indol reaction : A faint red color is produced in pepton-water culture 
(for 24 hours at 25^C), w'hen it is warmed with addition of HoSO^ or HCl. 

Acid : It produces acids in a solution containing glucose. 

By these properties this bacillus is pro\-ed to be the coli-bacillus. 

TJic I\Iicrococcus I. 

Form : The cell cultured in bouillon for 24 hours has a diameter of 
about o.S/i, and appears usually in the form of diplococcus. 

Gram's Method : Positive. 

Oxygen: Growth is better in presence of air. 

Bouillon : A little white precipitate was formed, when cultured for 
2 days at 2 3°C. No scum was formed, although kept for more than 2 days. 

Gelatine plate : A yellow, round, sharply defined, moist, bright, homo- 



4IO S. Sjiivamura. 

genous and elevated colony, that docs not liquefy gelatine, is formed on 
the surface. By weak magnification the appearance is the same. Deep 
colony is a white point. 

Gelatine streak : Colony is homogenous, and at first white but after- 
wards turns yellowish brown : It does not liquefy gelatine. 

Gelatine stab-culture : Colony is formed straightly along the inocu- 
lated line to the bottom. 

Agar streak : Colony is elevated, homogenous, moist, and at first 
white but afterwards assumes a faint brown. 

Potato : A white, homogenous colony is produced along the inoculated 
line in 6 days at 2 3'-'C. 

Milk : Milk is coagulated, acid reaction being produced. 

Gas-production : Gas is not evolved. 

HgS : HgS is not formed. 

Reduction : Nitrate is reduced to nitrite. 

Acid : Acid are produced when cultivated in a nutritive glucose 

bouillon. 

» 

TJic j\Iicrococciis II. 

Form : The cell cultivated in bouillon for 24 hours is about 0.8/i in 
diameter. It occurs usually in the form of diplococcus, but sometimes four 
are united. 

Grain's method : Positive. 

Oxygen : Aerobic. 

Bouillon : At I5°C on the fourth day of inoculation it becomes turbid, 
and on the sixth day a precipitate is formed, tlic supernatant fluid remaining 
clear. It is the same after 20 day's culture. 

Gelatine plate : Surface colony is round, convex, sharply defined, 
homogenous, moist, bright, white antl has a porcelain lustre. Gelatine is 
not liquefied. Weakly magnified appearance is the same as the above. 
Deep colony is a white point. 

Gelatine stab-culture : Colony is formed straightly to the bottom along 
the inoculated line. 



Investigations on Fljicherie. 411 

Gelatine streak : A white, moist, homogeneous, elevated colony is 
formed along the inoculated line, without liquefying the media. 

Agar streak : A white, moist, homogeneous, elevated colony is formed 
which extends very soon the whole surface. 

Potato : A white, homogeneous, moist, elevated colony is formed on 
the fourth day of inoculation at 30°C. 

Milk : Milk is coagulated, acid reaction being produced. 

Gas-production : Gas is not evolved. 

HgS : HgS is not formed. 

Reduction : Nitrate is reduced to nitrite. 

Acid : Acids produced by cultivating in a nutritive glucose solution 
(with Ca CO3) for 6 days at 36°C. was found to be 0.12^ calculated from 
the dissolved Ca O as lactic acid. 

TJie Micrococcus III. 

Form : The cell cultured in bouillon for 24 hours is about i /u. in 
diameter. Usually two are united but sometimes four. 

Gram' s method : Positive. 

Oxygen : Aerobic. 

Bouillon : It becomes turbid by two day's cultivation at 23^C. After 
20 days a feeble scum is formed, and a yellow precipitate on the bottom, 
the supernatant fluid remaining still turbid. 

Gelatine plate : Surface colony is yellow, round, convex, sharply 
defined, moist and bright. By weak magnifying power the appearance is 
the same, granular consistence being visible. Gelatine is liquefied. Deep 
colony is a white point. 

Gelatine stab-culture : Colony is formed straightly to the bottom along 
the inoculated line, liquefying it in the shape of a nail. 

Gelatine streak : A sulphur-yellow colony is formed along the ino- 
culated line, liquefying it completely after a few days. 

Agar streak : An elevated moist homogeneous colony is formed, the 
color of which is white at first, but turns sulphur-yellow after a few days. 

Potato : A very elevated, moist, bright, homogeneous colony is formed 



412 S. Sawamura. 

along the inoculated line. It is yellow at first, but turns brown after a few 
days. 

Milk : Milk is coagulated, acid reaction being produced. 

Gas-production : Gas is not evolved. 

HgS : HgS is not formed. 

Reduction : Nitrate is reduced to nitrite. 

Acid : Acids were produced by cultivating in pepton-water containing 
glucose. 



In the intestinal juice other micrococci and bacilli are of course present, 
although their number is commonly less than the above described. But as 
the colonies formed by those micrococci which arc chromogenous, are at 
first white and assume the proper tint after many days of culture, mistakes 
are possible by not giving time enough 



Tlic Micrococcus present in the Eggs. 

Form. The cell cultivated in bouillon for 24 hours is i [x in diameter. 
It occurs always in packet-form in nutritive fluids. But in the intestinal 
juice of the larvae it occurs in the form of diplococcus. 

Gram' s method : Positive. 

Oxygen : Aerobic. 

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, elevated colony is formed along the in- 
oculated line, gelatine being quickly liquefied. 

Gelatine stab-culture : Colonics 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. 



Investigations on Haclierie. 4^3 

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. 

HgS : HoS is not formed. 

Reduction : Nitrate is reduced to intritc. 

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 Sarciua lutca Fli'iggc. 



Chapter III. 
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 
of the micrococci commonl}' found abundantly in the diseased larva. 

Expcriniciit I. 
This experiment was performed in this College in October of 1901. At 



414 S. Sairamiira. 

this time it was rather cold and since flacheric 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 II cultured 
in bouillon for 9 days at 36°C. The filtrate was prepared from the above 
culture by filtering through Cliauihcrland' 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 larvai of the second 
day of the fourth age. The number of the larvae used for each experiment 
was 20, and the quantity of the materials used w^as 1,5 cc. to 100 grs. of 
mulberry-leaves. The larvae showed a very good appetite, and then they 
were treated as usual. The temperature in the box was 2i^C. 

Oct. 30. When they were examined in the morning, there were no 
diseased larvae found. Hence the culture, the filtrate and the heated liquid 
were given again to the larvae as before, and the temperature was raised to 
25°C. and water was bcsprinckled in order to increase moisture, because high 
temperature and moisture arc favorable to the development of flacheric. 
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 tlie sections. 

Nov. I. All the larva), except one in the control experiment that had 
died, span healthy cocoons. An excreta of the larvaj 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 II. 

The negative result obtained in tlie former cxiierinient might have been 
due to tlie low tem])erature. So this exi)eriment was perfornUHl to repeat 






Inyestigations on Flaeliei ic. 



415 



the former one using higher temperature. The culture used for this was also 
that of Micrococcus II cultured in bouillon for 3 days at 36^0. Filtration 
and heating \vcrc performed as in the former experiment. 

Nov. 8. In the afternoon the materials were given twice respective- 
ly to 20 of the larva; of the fourth day of the fourth age in the same manner 
as in the former. The intestinal juice of the diseased larvae was also 
given to 10 larvjE. At 4 P.IM. they were put in a 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 larva; soon got 
into the stage of ecdycis. and on loth they ended ecdycis. From this da}' 
on death took place. 

Nov. II. The silk-worms were transfered to a room of 21-C. 

The number of the dead larva; will be seen from the following table. 



Date. 


Control. 


The culture. 


The filtrate. 


The heated 
cultui-c. 


The intestinal 
juice. 


Nov. 10 


I 


4 


I 





7 


II 

















12 





I 





I 





13 





I 








2 


14 







I 








15 














I 


16 

















17 


I 





2 








iS 

















19 

















20 

















21 


I 


4 











Total 


3 


10 


4 


I 


10 







The remainder ftirmed coccooiis. 

As soon as the larv;v died, their intestinal juice was examined with a 



4i6 



S. Saivamnra. 



microscope, and according to the microbes present in the juice and also 
other symptoms, the disease of the dead larvae was grouped as follows: — 





Control. 


Culture. 


Filtrate. 


Heated cul- 
ture. 


Intestinal juice. 


Flacherie' I ... 

II 

Grasserie 


o 

I 
o 

2 


7 
o 

I 


I 
o 

I 
3 


o 
o 
o 

I 


7 

I 

o 


rebrine 


O 






Total 


3 


lO 


5 


I 


lO 


Flacherie in % of total 
larvre 


5 


35 


5 


— 


8 







Contrary to the former experiment many fiacheric-paticnts were pro- 
duced in this. It can, therefore, be concluded : — 

(I), 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 ; 

(11) that pathogenic action is not due to the production of toxin. 



Experiment III. 

This experiment was performed to confirm once more the result of the 
former ones. The cultures used in this experiment were prepared from 
Micrococcus II cultivated in bouillon for lo days at 36^C. and from micro- 
coccus I 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 
larva; on the second day of the fifth age, taking 20 larvce for each ex- 
periment. The temperature of the room was I5°C. and moisture 54. 
They were kept to the icSth, no diseased one being observed. They were 
tlierefore placed in a thermostat and kept at 27"C. and on the 22nd they 

> Machcric I dcnoLcs that in wliich micrococci were aliundant, and llaclieric II wlici'c l)aciili were 
abundant. 



Investiffations on riaclieric. 



417 



were again transferee! to the former room, and on the 26th they formed 
coccoons. 

The number of the dead larvae during the experiment was as follows : — 



Date. 


Control. 


Culture of 
Micrococcus I. 


Culture of 
Micrococcus 11. 


The filtrate. 


Nov. 19 
20 
21 

23 
24 
25 


3 

I 
2 



I 


I 

2 
2 
I 


4 


2 

I 
o 
o 
o 
o 
o 


I 

2 
O 

o 
o 

I 


Total 


7 


10 


3 


7 



The disease was grouped as follows : — 





Control. 


Micrococcus I. 


Micrococcus 11. 


Filtrate. 


Flacherie I 

„ II 

Grasscric 


3 

2 
O 
2 


5 
4 
I 
o 


1 

I 
o 
o 


7 




Pebrinc 









Total 


7 


lO 


3 


7 


Flacherie in ^y of the 
total larvn: 


:25 


45 


15 


^ - 







It \\\\\ be seen from these tables that tlacherie was more in the larx'o: 
that were not infected artificial]}-, tlian in those that received the bacteria. 
From this fact it can be learned that the bacteria, that cause tlacherie, are 
already present in the vicinity of the lar\a: and even in their intestines, wait- 
ing for an opportunity for development. Since many patients appeared 
among the larvai fed with the filtrate, tlaclierie would seem to be caused by 



41 8 S. Sawamiira. 

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 ac^ainst the belief held by the scriculturists 
of the present day. 



Experiments IV. 

This experiment was performed to investigate once more the pathogeny 
of the micrococci. 

1902. May 8. Llicroeocciis II cultured in the decoction of mulberry- 
leaves for 7 days at 36^C. were given four times to 100 larvae {AoJiiki 
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 lar\-cC examined on the first day of the fifth age 
were as follows : — 

Healthy Dead Lost 

Control 89 6 5 

Inoculated 91 3 6 

The average temperature and moisture during the experiment were as 
follows : — 

Temjierature Moisture 

First age .i9,O^C. 68,4 

Second „ 21,5 71,4 

Third „ 22,0 75,3 

j'^ourth „ 20,0 78,1 

Experiment 1^. 

Since flacherie occurs usualK' more in old larvae than in young ones, the 
negative result obtained in the former experiments might be due to the fact 



Investijrations on Flaclteric. 



419 



that the bacteria were fed to young larva;. Therefore this experiment was 
repeated, using old larvae. 

The bacteria used for this experiment were Micrococcus II 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 65^0.^ 

IV. The same to which formalin was added in the proportion of i drop 
to 10 cc. of the culture.^ 

May 22. 3 P.M. The materials above described were given together 
with mulberry-leaves'^ each to 100 larva {Akaliiki 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. 


jMoisture. 


May 22 


22,5^C. 


70,0 


23 


22,0 


78.0 


24 


22 I 


80,0 


25 


21,1 


75,0 


26 


22,0 


82.0 


27 


22,0 


69.0 


28 


18.5 


74.7 


29 


-"» "» 


■ ;^^-7 



■J Sard 11(1 ///U-ii Fi'i'is^^i-. 

» Sterilizcil. 

3 Sterilized. 

■« 1,5 cc. of the materials to loo i^.?. of mulhcrrv-leave^ 



420 8. Sawamiira. 



Experiment VI. 

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.J\I. 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 larvae {Akahiki variety) of the second day of the fifth age, 
In the intestinal juice there Avere 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 17,0-C. 

29 18,0 

30 20,0 

31 19.5 
June I 21,5 

2 19,8 

3 20,9 

From these experimental results it is clear that silk-worms do not 
become ill from fiacherie when the surrounding conditions are favorable to 
their health, and tluy have resistance-power. These results agree with 
that reported by the Austrian Agricultural Experimental Station. 

lixpcriineiit J Y/. 

Sinci.- the negative results obtained in the former ex|)eriments nu'glit 
have been duv to the insufficiency of tlu' number of the bacteria, a fiu'tlier 



Investigations on Flaolit'iic. 42 i 

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 : — 

Micrococc2is I. 

II- 
III 
Coli-bacillus (isolated from the diceased larva). 
Bacillus mcscntcricus vulgatits Fli'iggc. 

,, ,, fnscus ,, 

Bacillus subtilis Colin. 
June 7. 2 P.M. The above materials were injected into the larva; 
(^<?/^/-^/-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 larv^ 
were selected from each section, and at the same time 100 larvae 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 naturall)' no decisive 
result. The results of the injection must be observed within 3 or 4 days. 
The results after three days were as follows : — 

I. Distilled Water. 

The larva; 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 app^tit-\ On the afternoon of the second day four 



422 S. Sawaimira. 

of them died of flacherie ; in the intestinal juice micrococci and Bac. incga- 
tJicrium 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 II. 

The larvae lost appetite. On the afternoon of the second day one died 
of flacherie, in which much of Bac. incgatJicriuin and little of micrococci 
were found. On the second night three died, in which a great number of 
micrococci was found. 



4. Micrococcus III. 

The larv^a; lost appetite. On tliat night two died of flacherie, in one 
of which the micrococci prevalent, while in the other Bac. mcgatlicriuni 
exceeded the number of micrococci. On the second day five died of flache- 
rie, in A\'hich a great number of micrococci was found. 



5. Coli-bacillus. 

The larva,' lost appetite completely. On the afternoon of tlie 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 da}- one died. 
In the former only coli-bacilli, while in the latter Bac. incgat/icriinii was 
found. 

6. Bacillus vicsciitcricus vulgatus Fliiggc. 

The larvcc lost appetite completely. On the afternoon of the second 
day two died, in one of which Bac. nics. vulgatus prevailed, w hile in the 
other micrococci were more abundant. On the second night se\en died, in 
six of which llac. incs. 7'ulgatus, but in one only micrococci were observed. 



Investigations on Flacherio. 



423 



7. Bacillus incscntcricus fusciis Fliigge. 

The larvae lost appetite. On the second night seven died of flacherie, 
in four of which onjy Bac. mcs. fusais, but in three this microbe together 
with micrococci were found. On the afternoon of the third day one died, 
in which Bac. mcs.fusctis alone was abserved. 



8. Bacillus subtilis Co/in. 



The larvcE 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. 


in )>(,' of the larvre 
taken for the test. 


Control 














Water 








5 


2 
2 


20 


Micrococcus I 


70 


// 





4 





40 


/// 


2 


5 





70 


Coli-bacillus 






9 
9 


I 




100 


Bac. mes. viilgatiis 


90 


„ „ ftisciis 





7 


I 


So 


Bac. subtilis 


I 





5 


60 



From the results obtained in this experiment the following conclusions 
can be drawn : — 

I. Many species of bacteria can propagate in the intestinal juice of the 
larvx and cause flacherie. 



424 S. Saw.iin'.ira. 

2. Disorder in tlic digestive organ such as injection of water causes 
flacherie. 

3. From the above facts it is clear that bacteria, that" can cause flache- 
rie, arc present at all times in the intestinal canal of the larvae 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 frequent!)' than Avhen 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 Micro- 
coccus III cultured in a decoction of mulberry-leaves in absence of air for 
7 days at 36'- C. 

It was filtered through CJianibcrlana s filter and a part of it was neu- 
tralized with Nag CO 3. 

June 9. 2 r.M. 0,05 cc of the original and the neutralized filtrates 
w'ere injected into the intestines of the larvae {AoJiiki variety) on the fourth 
day of the fifth age. The results were as follows : — 

I. TJic Oric^inal Filtrate. 

Eleven larvae which received this material remained inactive for two 
hours. One of them was killetl and the intestinal juice was examined, in 
which Bac. iiu\i^atheriinii was found in large number. On the next morning 
eight larvaj chtd, in two of which micrococci abountlcd, but little of Bac. 
incgatlicrium was present; and in three others Bac. mcgatJicriuin abounded, 
while micrococci were few; while in three others only Bac. vicgatJicriiim 
was found. On the nth two died, in which 7?^?^:. wr^.'-^/Z/r/vV/;// abounded, 
but few micrococci were found. 



Investigations on Flacherie. 



425 



2. TJic Ncutrali'jcd Filtrate. 

On the forenoon of the loth nine out of twelve larvae used for the opera- 
tion died, in which Bac. mcgathcrunn abounded. On the i ith three died, in 
which both Bac. megatJicriinn and micrococci were found. 

Experiment IX. 

Since some bacteria produce a powerful toxin only when they are 
mixedly infected, 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. II P.M. 0,1 cc. of the original and neutralized filtrate of the 
above cultures were injected into the larvae {Aoliiki \7\.x\QXy) on the fifth day 
of the fifth age. 

The number of the dead was as follows : — 





Number of 

the larva; 

tested. 


First day. 


Second day. 


Third day. 


Fourth day. 


% of the 
dead. 


Water •.. 


00000 


00000 



6 

4 
6 

5 


2 
4 
4 
4 








CO 


Filtrate from coli- 
bacillus 

The same neutralized. 

Filti'ate from the mix- 
ed culture 


I CD 
ICO 

100 


The same neutralized. 


00 



According to the kinds of the bacteria found in the intestinal juice they 
may be grouped as follows : — 



Water 

l*'iltrate from coli-bacillus. 

The same neutralized ... 
Filtrate from the mixed 

culture 

The same neutralized ... 



Much Bac. 
fiic'gal/it. riiiin. 



/nil. incgatlwrtum Bac. inc^at/urintn 
-f miciococci. -i- coli-bacillus. 



Fac. vtcc^itficTiinii 
-(-coli-bacillus 
-(- micrococci. 



426 S. Sawamiira. 

As many died of flachcric 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 flacherie 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 tlierc 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 cnzyms of silk- worm 
are active only in an alkaline solution. The micrococci found in the diseas- 
ed larva; and that in the eggs as Axell as Bae. mcgatlicriitiu, Bae. eoll, Bac. 
siibtilis, Bae. vies, viilgatiis and fnsens, all produce acids in a solution con- 
taining carbohydrates, which were comfirmcd 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 b\' the bacteria. 
1 lence the effect of injection of acids was studied. 

0,1 cc. of distilled water, 2)% normal sodium carbonate solution, 2% 
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. Wy this operation some vomited fluid, esiieciall}' man_\' of those 
injected witli water and srxlium carbonate solution. All the larwx seemed 
somewhat inactive, but those injected w ith water anil soilium carbonate solu- 
tion showed very gootl appetite after a few hours. - 

* Compare .nl>o the altovc cxinriiiicnt. y. 420. 

-' Tlic larviv injected witb cllstillcd water did not die within 24 lioinr. 



Investigations on Flacherie. 4^7 

Those injected witli the acids died with vomition and diarrhea after 
about lo 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 o, I cc. of iO% 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 ol 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.^ 



Experiment XI. 

But the bacteria seem to produce a certain poison, although it is no 
toxiii. 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 
Bae. niei^atlierunii and the micrococci were also proved by the writer. - 

This experiment was performed therefore to observe the etlect of nitrite 
on the silk-worm. 

July 4. 9 A.M. 20 larvie of the second day of the fifth stage were 
fed with nudberry-leaves moistened with a io^q^ solution of sodium nitrite 
for a day.-^ On the next morning a lar\a ilied. 

1 In higher animals also llic sccrclio;i if the iiitcsliiial juice is much accelerated by presence of 
acids. Btitige. Tliysiol. Chcmie. 

2 Mulbcny-lcavcs contains ol'ten much nitrate. 

3 1.5 cc of tlic solufon to ico trr?. of the leaves. The larva- diil r.ot cat the leaves as usual. 



4^8 S. Saiva III lira. 

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 o,i cc. of i% solution of sodium nitrite in the usual manner, 
six out of seven larvae used for the experiment died instantly, the bodies 
of which were softened and stretched. Those to which only 0.05 cc. were 
injected, \vere, 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 larvae 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. 



Jixpcr'uncnt XII. 

Since Bac. DicgathcriiDn or Bac, call are of general occurrence it is no 
wonder tliat they propagate also in tlie intestines of the silk-worms. But 
as to the micrococci it is different. 

As a micrococcus and Bac. mcgathcriiiin exist in the interior of some 
eggs, they might come from the eggs as Pasteur and Macc/iiati 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 fount! in the eggs was quite different from those 
usually found in the diseased larVcU. 

1902 June 23.' Agar-plates were infected with small fragments of a 
mulberry-leaf 

Tlie Cfjlonies formed after two days were as follows : — 

The original plate : Colonics of a large bacillus {Bac. mcgat/icriiim 

or Bac. subtilis ?) 
The second diltition: Xuiiktoiis colonirs of the large bacillus and 
micrococci. 

1 It laiiicd Iwo (l.iys he-tuix-. 



Investigations on Flachorie. 



429 



The third cHlution : Colonics of the hirge bacilhis and white colonics 
of micrococcus. 
June 24. The former experiment was repeated, as on the previous daj' 
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 colonics of micrococcus. 
June 27. The experiment was repeated with mulberr)'-leaves oi Hara- 
jiikit wdierc silk-worms were never reared before. 
The results were as follows: — 

The original plate : Colonies of various bacteria covered tlic 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 wdiether 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.]\I. The micrococci isolated from mulbcrrj'-leaves and 
those of the diseased larvic were inoculated in the usual manner into the 
larvse of the fifth day of the fifth stage, and as they died, their intestinal 
juice was examined with a microscope. 

The results wxre as follows : — 





Kuniher 
of larvrc. 


Fii-st day. 


Second 
day. 


Third 
day. 


Total. 


% of the 
dead. 


Water 


10 
10 
10 
10 
10 
36 




2 





I 

5 
S 

4 

5 



2 
2 


6 
5 



3 
7 

10 
10 
10 




Micrococcus II of silk-uonn 

Bac. viegatheritivi from sillc-wonn. 
Micrococcus I from mulberry- 
leaves*- 


70 
100 

IC» 


Micrococcus 11 from mulberry- 

leaves^ 

Control 


100 







Micrococcus I from muIberry-lcavcs formed a white colony, while Micrococcus II a light brown 
colony on agar. 



430 



S. SawauuiiM. 



The dead larvae were grouped according to the species of the bacteria 
found in the intestines as follows : — 



Micrococcus 
only. 



Bac. megath 
erium and 
micrococci. 



Bac. megath- 
Bac. nu'gatJi-^ ^/;^;;^ ^nd 
erium only, coli-bacillus. 



Very few 
bacteria. 



Water 

Micrococcus II of silk- worm. 

Bac. megatherium 

Micrococcus I from mul- 
berry-leaves 

Micrococcus II irom mul- 
berrv-leaves 



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 larvai exist also on mulberry-leaves or not, micrococci were 
isolated from these leaves and their properties were examined.^ 



No. 



Form : The diameter of the cell cultivated in bouillon for 24 hours is 
I /i. Commonly two arc 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 tlie seventh day a white ring is formed on the wall of 
the tube and a white jjrecijMtate is formed, the su]X'rnatant Huitl becoming 
clear. i\fter 20 days a feeble scum appears. 

rrelatine plate : Surface colony is white, round, sharply tlefuied, lipped, 

' The leaves came partly from the College farm in K'oiiwlni, and \ artly from a garden in Tokio 
where no silk-worms were kejit for 30 years. 



iivestigations on Flacheric. 43 ^ 

moist and has porcclain-likc 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 I2°C. after 20 day's culture, colonics 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 1 5"C. 

H^S : H^S 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 
S% of glucose. This micrococcus is therefore probably Micrococcus corona- 
tiis Fli'iggc, 

No. 2. 

Form : The diameter of the cell cultured in bouillon for 24 hours is 
0.8 /i. Usually two are waited. The microbe is colored by Grani s method. 

Bouillon : On the fifth day it becomes turbid, and on the 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 tiays 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 tleep brown. Gelatine is slowly licjuefied. 

Gelatine stabculture : Thread-like growth to the bottom and liquefac- 
tion in the form of a nail. 

i\gar plate : At 30'C the surface colon-^" is light brown moist, bright, 



432 S. Saivamiiru. 

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 a few days a yellow color. 

Potato : On the second day a flat, dr)-, light brown, granular colony 
along the inoculated line. 

]\Iilk : It is coagulated showing alkaline reaction. 

Oxygen : i\erobic. 

Gas : Gas is not evolved. 

HgS : HgS is not formed. 

Indol reaction : Faint reaction. 

Reduction : Nitrite is formed from nitrate. 

Acids : Acids are formed in glucose solution. 

This micrococcus is ]\Iicrococcus hicolor, Zimmcnnann. 

No. 3. 

Form : The cell cultured in bouillon for 24 hours has a diameter of 
0.8 //. Two are usually united, but sometimes four, isolated cells are rare. 
It is colored by Gravi s method. 

Bouillon : 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 folate : Surface colony is dirt}' white, round, convex, with a 
white ring. By weak magnification, the centre seems deeply colored and it 
becomes lighter towards the margin. Gelatine is slowly*liquefied. 

Gelatine streak : A light brown, homogeneous colony is formed, gelatine 
l)ei ng liquefied. 

Gelatine stabculture : Thread-like growth to the bottom, licjuefying 
gelatine along the inoculated line. 

Agar streak : INIoist, homogeneous, tenacious colony which is white at 
first, but l:)ecomes light redilish brown afterwards. 

Potato : It tiid not grow on potato in a week at 30^C. 

Milk: Milk is coagulated, aciils being formed. 



I 



Investigations on Flachcrie. 433 

Oxygen : Aerobic. 

Gas : Gas is not formed. 

HgS : HgS is formed. 

Reduction : Nitrate is reduced to nitrite. 

Acid : Acid is formed in glucose solution. 

No. 4. 

Form : The cell cultivated in bouillon for 34 hours has a diameter of 
I fx. Two are usually united. It is colored by Grani s method. 

Bouillon : At 2 3^C. on the second day a white precipitate is formed. 
Scum is not formed by 20 days' culture. 

Gelatine plate: Surface colony is yellow, moist, bright, round, sharply 
defined, convex and homogeneous. The appearance is the same by weak 
magnification. Deep colonies appear as white points. 

Gelatine streak.: Homogeneous colony along the inoculated line, the 
color of which is white at first but turns yellowish brown afterwards. Gela- 
tine is not liquefied. 

Gelatine stabculture : Thread like growth to the bottom. 

Agar streak: Elevated, homogeneous, moist, dirty-white colony. 

Potato: At 23'~'C. on the sixth day of inoculation a white elevated 
homogeneous colony is formed along the inoculated line. 

?ililk: Milk is coagulated, turning acid. 

Oxygen : Aerobic. 

Gas : Gas is not formed. 

IIoS: MoS is not formed. 

Reduction : Nitrate is reduced to nitrite. 

Acids : Acids are formed in glucose solution. 

This micrococcus is the same as Micrococcus /of the silk-worm. 

No. 5. 

Form : The cell of 24 hour's culture in bouillon has a dianutor of 1.2 /'. 
Two or more are united. It is colored by Gram' s method. 



434 S. Sawamiira. 

Bouillon : At 2 3^C. on the second day it becomes turbid and on the 
fourth day a scum is formed and on the seventh day a white precipitate ap- 
pears, the middle part being clear. It remains the same after 20 days. 

Gelatine plate : Surface colony is yellow, round, sharply defined, 
granular, moist and bright. It appears alike by weak magnification. Gela- 
tine is liquefied. Deep colony is a yellow point. 

Gelatine streak : Sulphur-yellow, moist, homogeneous, elevated colony 
is formed along the inoculated line. Gelatine is quickly liquefied, yellow 
precipitate being formed. 

Gelatine stabculture : Thread-like growth to the bottom, gelatine being 
liquefied at first in the form of a funnel but afterwards cylindrically. 

Agar streak : Sulphur-yellow, moist, homogeneous, devated colony 
along the inoculated line. 

Potato : ' At 23^C. dr}-, flat, white colon)', having }'ello\v granules there- 
on. 

Millk : Milk is coagulated, acids being formed. 

Oxygen : Aerobic. 

Gas : (jas is not evolved. 

HoS : HoS is formed. 

Reduction. Nitrate is reduced to nitrite. 

Acids : Acids one formed in glucose solution. 

The yellow pigment is insoluble in water or alcohol, but soluble in 
potash solution. The pigment dissolved in the latter solution becomes color- 
less by the addition of HCl, which is restored again to yellow by alkali. 

This micrococcus is a variety of I\Iicrocpccus Intciis, LcJunann ct Xcu- 
inaiui. 

No. 6. 

I'orm : The cell cultured in bouillon for 24 hours has a dianuter of 
0.8 /i. Commonly two are united. It is colored by (Irani s method. 

Bouillon : On the third da}' a precipitate appears, and after 20 days 
a feeble yellow-brown scum and a }-ellow-brown precipitate are fornu il. 

(iL-latinu plate : .Surface colony is yellowish brown, moist, bright, round. 



IiiYOstisations on FLicliorie. 435 

sharply defined and elevated. By weak mag-nification rri-anular consistence 
is seen. Deep colony is a wliite point. 

Gelatine streak : Light brown, tenacious colony is formed along the 
inoculated line. Gelatine is liquefied, film being formed. 

Gelatine stabculture : Thread-like growth to the bottom, liquefying 
Gelatine in the form of a funnel. 

Agar streak : Yellowish brown, moist, homogeneous, elevated colony. 

Potato : At 23^C. on the second day a yellow colony is formed along 
the inoculated line. It assumes gradually a reddish yellow color and be- 
comes elevated, dry and granular. 

Milk : Milk is coagulated, showing an alkaline reaction. 

Oxygen : Aerobic. 

Gas : No gas is evolved. 

HgS : HgS is formed. 

Reduction : Nitrate is reduced to nitrite. 

Indol reaction : A faint reaction. 

Acids : 0.024^5^ of acid, calculated from the dissolved Ca O as lactic 
acid, were formed in pcpton-watcr containing 5J^ of glucose for a week at 
15-20'^C. 

This micrococcus is probably Micrococcus pyogenes aureus, LcJniianii 
ct Neumann. 

No. 7. 

Form : The cell cultivated in bouillon for 24 hours has a diameter of 
0.8 IX. Usually two are united, but sometimes four. It is colored by 
Grain's mcthotl. 

Bouillon : On the second da\' it becomes turbid, and after 20 days a 
feeble ring on the wall, and a )-ellow precipitate were formed. 

Gelatine plate : Surface colony is yellow, moist, bright, round, convex 
and sharpl)' defined. By weak m;ignification it appears granular. Gelatine 
is liquefied. Deep colonies appe;ir as white points. 

Gelatine streak : Sulphur-yclKnv. homogeneous colony is formed along 
the inoculated line, gelatine being liquefied very quickly. 



43^ S. SaTramiira. 

Gelatine stick: Thread-like growth to the bottom, liquefying' gelatine 
in the form of a nail. 

Agar streak : An elevated homogeneous, moist white colony is formed 
which gradually turns yellow. 

Potato : At room-temperature elevated, yellow, moist bright, homo- 
geneous colonies are formed along the inoculated line. 

Milk : Milk is coagulated, acids being formed. 

Oxygen : Aerobic. 

Gas : Gas is not formed. 

H'jS : H„S is not formed. 

Reduction : Nitrate is reduced to nitrite. 

Acids : 0.013^ of acid, culculatcd from ^thc dissolved Ca O as lactic 
acid, is formed by cultivating in pepton-water containing glucose and Ca 
C03 for 14 days at i5-2o°C. 

The yellow pigment is insoluble in water or alcohol, but soluble in 
potash solution. The color is not destroyed by IICl or H^SO^. 

This micrococcus is the same as Micrococcus III of the silk-worm, and 
is probably Streptococcus bombycis of Macchiati. 

No. 8. 

Form : The cell cultured in bouillon for 24 hours has a diameter of 
1 fx. Commonly two are united but sometimes for.r. It is colored by 
(i rain's method. 

l^ouillon : At 1 5^C. on the fourth day it becomes a little turbid, and on 
the si.xth day a precipitate settles. It remains the same after 20 days. 

Gelatine plate : Surface colony is white, round, elevated, sharply de- 
fuied, and moist. V>y weak magnification it appears homogeneous. 13eep 
colonies appear as white points. 

(ielatine streak : An elevated, white, nio'st. liomogeneous colony is 
formc-d. 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. 



luvestigatious ou Flaclieric. 437 

Potato : An elevated, white, moist, bright, homogeneous colony. On 
the central line it is more elevated. 

Milk : Milk is coagulated, acids being formed. 

Oxygen : Aerobic. 

Gas : Gas is not formed. 

HgS : HgS is formed. 

Reduction : Nitrate is reduced to nitrite. 

iVcids : Acids are formed in glucose solution. 

This is the same as IllicroLOcciis // of the silk-worm, and is probably the 
Streptococcus Pastorianus of Krassilschtschlk. 

No. 9. 

Form : The cell cultivated in bouillon fur 24 hours is somewhat oblong 
and 0.8 yu. in the longer diameter. Usuall}- two are united. It is coloured 
by Grauis 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 dr}'. homogeneous colonx', which 
is much elevated on the central line, (ielatine is not licjuefied. 

Gelatine stabculture : Thread-like growth to the bottom. 

i\gar streak : It is the same as on gelatine, but the color is fainter. 

Potato : At 2 3''C. on the fourth da}- flat, light \-ellow. moist, homo- 
geneous colonies are formed along the inoculated line. 

Milk : Milk- is coagulated, acids being formed. 

Oxj'gen : Aerobic. 

Gas : Gas is not evolveil. 

II^S: I r,S is formed. 

Reduction : Nitrate is retluced to nitrite. 

Indol reaction : A flint reactit^n. 

Acids : o.Oi j?o of acid, culculatetl as lactic acid, w as produced in pepton- 

water containing ^^^ of gluc«-»se and some Ca,CO.. fi)r 14 da\-s at i5-'-20 C. 



438 S. Sawamurji. 

This micrococcus is Micrococcus aiirantiaca, Colin. 

No. lo. 

The colony of this micrococcus is dark purple. The properties are not 
yet examined minutely. 

Experiment XIV. 

Nov. II. 1902. On 10 A.]\I. 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 larva; 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 larva; and the temperature during the experi- 
ment were as follows : — 





Numljer 
of the 
larvce. 


II 


12 


13 


14 


15 


16 


17 


18 


19 


20 


21 


22 


23 


24 


25 


Total. 


% of the 
dead. 


1 


iiioriiing 


14 


II 


II 


iS 


16 


II 


9>5 " 


20 


13 


18 


20 


20 


20 


20 






Teiiiperaturc (c)iH 






































( 


evening 


22 


— 


— 


24 


20 


13 


— 


18 


— 


18 


25 


20 


20 


20 


20 






Control 


15 












































2 


2 


13 


Water 


10 











2 


2 














I 


I 














6 


60 


No. I 


10 











3 














1 





I 


2 











7 


70 


No. 2 


10 














5 


























No. 3 


9 











I 


4 


I 


I 














2 











9 


100 


No. 4 


10 











I 


6 





I 











I 











I 


10 


100 


No. 5 


10 





I 


2 


I 


4 


'> 




















10 


100 


No. 6 


S 











2 


3 


I 














I 


I 








S 


100 


No. 7 


10 








I 


I 


2 


3 


I 








2 












10 


100 


No. 8 


10 














I 


3 











2 


2 


2 








.0 


100 


No, 9 


10 











3 


2 


4 











I 












10 


100 


Micrococcus fidin 
llii^fRRs 


10 











4 


5 


I 




















10 


100 



Tcnipcralure afler the lylh was very low during the nigiil, as llie stove \\as not used at niglit. 



luvesligalious on riaclicric. 439 

The conditions of the hirvcc in each section of the experiment were us 
follows : — 



Control. 

15 silk-worms were kept for control. They were healthy and span 
cocoons on Nov. 25. Durini;- spinninij 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 larvaa 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-leaxTs in the intestines 
were green ; here only large bacilli were found. 

Nov. 15. On the morning one died, body softened and stretched out. 
Large bacilli were numerous, and also some diplococci were found. On the 
evening another died, bod)' expanded in the middle part. The pieces of 
mulberry-leaves in the body were brown ; streptococci were numerous. 

Nov. 20. On the morriing one died, body softened, back-part black. 
No fragments of mulberr}'-leaves present ; some diplococci in the intestines. 

Nov. 21. On the 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 0.41 1 gr. 

No. I. 

. After the operation the larvi\: lost appetite considerabl}-. 
Nt)\-. 14. On the morning one died, the third and fourth segments 
elongvited, excretion i>l soft brown ilunu" of a laint acid reaction. The fratj- 



I 



440 S. Saw.iumra. 

mcnts of mulbcrry-lcavcs 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, bod)' softened and expanded. 
The intestinal canal was filled only with lic^uid with numerous diplococci and 
few bacilli. 

Nov. 22. On the morning two died, one with swollen segments and 
black back-i)art. 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 larvae span cocoons, the average weight of 
which was 0.390 gr. 

l^y preparing a plate-culture from the intestinal juice of the diseased 
larva many white colonies of the micrococcus inoculated besides various others 
ai)peared. 

No. 2. 

The larvre lost ap})etite after the operation. 

Nov. 13. On the morning fi\'e 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 leaf-fragnK'uts i\.\u\ luuiKrous diplo- 
cocci and some streptococci. ihe third also with folils, l)ro\\n leaf frag- 
ments, numerous diplococci and some streptococci. The fourth resembled 
tlie third ; streptococci wtvc more numerous, while with the fifth iliplococci 
again prevailed, all the other conditions being the sanu-. 

Nov. 15. 'ihe remainder wen- unfortunate])' lost b)' an accident. 

Hy preparing an agar-plate fi'oni the intestinal juice of the dead larva 
many c(jlonies of the micrococcus inoculated were formed. 



Investigations on Flaclierie. 44^ 



No. 



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 ; dijDlococci 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. On the morning one died, body stretched and leaf fragments 
green, diplococci numerous. All the other larvae excreted liquid feces. 

Nov. 17. On the morning one died, fore-part of body somewhat trans- 
parent, back-part thin. The few leaf fragments green, few diplococci and 
pebrinc-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 cmpt\- inte- 
stines, the back part of the latter larva was black. 



No. 4. 

The larvae 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. On the morning three died, fore-part of bodies shrunk. Leaf 
fragments brown, numerous diplococci. On the evening three died, bodies 
softened, leaf fragments green, diplococci riumerous. 

Nov. 17. On the morning one died, bod\- softened, leaf fragments 
green, streptococci numerous. 

Nov. 21. On the morning one died, leaf fragments compressed; no 
bacteria were observed. 



I 



442 S. Sawniimr;i. 

Nov. 25. One died, body softened. Few fraji^ments of leaves, manj^ 
diplococci. 

By preparing an agar-platc from the dead larva numerous light brown 
colonics of diplococci were formed. 

No. 5. 

The larvse 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. 

No\^ 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. On the morning four died, bodies stretched and containing 
brown leaf fragments. In the first the back part of the body black and 
streptococci, numerous. In the 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 colonics of diplococci were produced in large number. 

No. 6. 

The larva- lost completcl}' appetite by the operation. 

Nov. 13. On the mording one- excreted a light yellow fluid of a fiintly 
acid reaction from the anus. 

Nov. 14. (^n the morning two died. One with stretched and softened 
body, and green leaf fragments. The second with shrunkiMi bod)' had ex- 
creted a yellow fluid of a faiutl)' aciil reaction from the anus. Leaf frag 
mcnts green, numerous diplococci present. 



Investigations on Flccliprio. 443 

Nov. 15. On the iTiorning two died, bodies faintK' }-ello\v. 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. 

Xo\'. 21. On the morning one died, body slirunk. 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. 



N( 



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, man)' 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 
l^resent. 

Nov. 16. rin-ee ilied. bodies ex[Kinded in the middle part. In the first 
leaf fragments green, large bacilli present. In the second the leaf fragments 
brown, streptococci and large l:)acilli numerous. In the third the leaf frag- 
ments brown, diplococci numerous. 

Nov. 17. On the morning one died, body softened am! leaf fragments 
brown and compressed, diplococci numerous. 

Nov. 20. On the morning two died ; one with softened and fainth- 
purple-colored bodies, leaf fragments brown and compressed, bacilli nume- 
rous. In the other body softened, the back part blackened, leaf fragments 
brown. Some sacc/idrojiiycitcs and large bacilli were found. 



444 



S. Sjnvaimira. 



Xo. 8. 

Nov. 15. On the niorning one diet), bod)' expanded in tlie niitldle jnirt, 
leaf fra_q;nients tureen, streptococci numerous. 

Xo\-. 16. On the morninq,- three died. In one the middle part of the 
faintly yellow colored bod\^ exi)anded, leaf fragments brown \vith man\' 
diplococci. In the second body softened, leaf fragments green and com- 
pressed, short bacilli numerous. In the third bod)' softened, leaf fragments 
green, numerous streptococci present. 

Xov. 20. ( )n the morning two died, with softened bodies and brown 
leaf fragments ; diplococci numerous; in one also bacilli present. 

Xov. 21. On the morning two died, one with contracted fore-part and 
a few di})lococci ; the (jther \\itli the fore-])art elongatetl and many 
diplococci. 

Xov. 22. On the morning two died diiring the spimiing of cocoon ; 
bodies softened, the back part black, leaf fragmen.ts green. In one bacilli 
\vere numerous but diplococci few ; in the other bacilli and dii)lococci were 
ccjuall)' nunierous. 

]-))• preparing aii agar-plate from the intestinal juice of the dead lar\'a 
(jnl)' white colonies of nn'crococci were formed. 

Xo. 9. 

Xow On the morning three died, two with the bod)' shrunk, and one 
with the botly elongated. One with man)' bacilli antl few dipk)Cocci, and 
brown leaf fragments; the second with numerous tliplococci, and green 
Compressed leaf fragments; the third with brown leaf fragments, numerous 
diplococci and some large bacilli. 

Xov. 15. On the morning one ilied, Ixnl)- black, leaf fragments brown, 
dii)l(Jcocci numerous but streptococci few. On the e\'ening one died, body 
softened, leaf fragments green, ch'plococci numerous. 

X''ov. 16. On the morning four dieil, bodies, softened. \\ ith three of 
them small 1)1. ick spots appeared on the boil)- ; intestinal contents were 



J 



Invcstij'atioiis on Flaclioric. 445 

f^rcen. In one of these (liplr)cocci, streptococci and some pebrinc-orsi^anisms 
were oljser\ecl. \\ itii tlie second lar^e bacilli and diplococci. With the 
third streptococci. With the fourth i^reen com[M'essed leaf fragments, and 
streptococci numerous, large bacilli few. 

Nov. 20. On the morning one died, bod)- s(^)ftened \^ith small black 
spots all over. Leaf fragments brown and compressed, diplococci numerous. 

By preparing an agar-platc from the intestinal juice of the dead larva, 
\'ellow colonies of di[)lococci were exclusixcly formed. 

MuTocoiTi/s frciii tltc co-gs. 

Nov. 14. On the morning two died, in one the body stretched, the 
second and third segments yellow, and leaf fragments brown and com[)rcss- 
cd, few diplococci were observed. In the other the bod\' was shrunk and 
hard, leaf fragments also brown and compressed ; sarcina was found cxclu- 
si\'el)'. On the e\'ening two died, leaf fragments brown and compressed, 
diplococci present. 

No\-. 15. On the morning fi\'e died, bodies 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, 
)'ellow colonies of sarcina were exclusi\'ely formed. 

1-^'om tlie results of these experimcMits the following conclusions were 
drawn. 

1. The micrococci on the mulberry-leaves can cause flacherie. Xo. 1 
Ix-ing the least able to multiiily in the intestines of the silk-worni. 

2. Flacherie can be caused b)' this micrococcus, what can be proved 
by the fact that b\' prei^aring a plate culture from the tliseased larva: the 
colonies of the inoculated micrococcus were formeil '\\\ greater number or 
exclusix'ch'. 

3. Since tlacherie is caused by injecting water into the intestines, it 
is clear that the Ixicteria that can cause tlacherie exist alwa\-s in the intes- 
tinal canal, which fact pnnes also th.\t the mulberry-leax'es are the carriers 
of the u'erms. 



44^' S. Sawrtiiiiira. 

4. The micrococcus in the eggs can cause flacherie. 

5. The decrease of appetite of the silk-worm by the injection of water 
or l^acteria can be obser\^ed also from the diminished weight of the cocoons 
formed. 

Axxrage weight of a cocoon. 

Control 0.487 gr. 

Water injected 0.41 1 

Micrococcus Xo. I inocuLated O.390 

6. Any constant relation between the bacteria inoculated nr.d the 
s\'mptoms of the malady was not observed in these experiments. 

7. W'hen mulberry-leaves in the intestines are green, the reaction of tiic 
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. iiicgatJicriuvi seems to multiply usually after the micrococci 
had developed juxuriantl)'. 

9. At low temperature the bacteria do not bring on flacherie very 
soon.'' Ihe cause will probabh' be due to the slow growth of tlie loacteria 
at the low temperature. 

Gciicfal ( ^onchisioiis. 

!• rom the rc:sults of the series of the c.xi)erimiMits abo\e described the 
following conclusions were drawn. 

I. There is no doubt that Hacheric is cause-d by the growth of bacteria 
ill the intestinal juice. 

II. The bacteria usualK' f )und in hirge number in the intestinal juice 
of the diseased lar\;e are \arious kinds of micrococci and two kinds of 
bacilh. In most cases micrococci only or together with few large l)acilli 
are lonnd. .\ short bacilhis is also usuall)- found along w ith the niicrococci ; 
liowextr cases in which the short bacillus alone is found are \'er)' rare. 
Hi'sides tlusi' microbes there- ari- man\' other kinds which exist in a small 
luiinbi r in tin- disease'd animals. 

' Coiiipare' l'"x|irriniciits \' 1 1 and \II. 



Iiivostisrtlioiis oil riiU'lioric. 447 

ITT. The larj^c bacillus was identified witli lutcillits mcg-atJicriiim, De 
Bary, and the short one with tlie co]i-])aci]lus. 

I\^ There exists in the interior of the cgi^s of the silk-worm usually 
a micrococcus and a lari^e bacillus. The former was identified with Sarcina 
hitca, Flif/^-o-c and the latter with naci/liis mcc^athcriinii, Dc Bary. 

V. Various kinds of micrococci usually adhere to the mulberry-leaves. 
The writer isolated from mulberry-leax'es lo species of micrococci. Nine 
of them were used for the experiments, by which it was decided that flachc- 
rie is caused by these micrococci and the sarcina isolated from the eggs. 
The micrococci isolated from the dead larvae were identified with those iso- 
lated from mulberry-leaves. 

\'I. It is clear that the sources of the bacteria, which multiph' in the 
intestines of the silk-worm and cause flacherie, are the mulbcrr\--lcaves ser- 
ving as food. But when the ]ar\ ,x are healthy they resist the action of the 
bacteria. However when silk-worms arc 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-lea\"es. 

\\\. Flacherie is, as abov^e explained, n(^t caused l)y an\- special bac- 
teria, hence MaccJiiatis' and Krassilschtsc/iik' s assumption can not be corn- 
firmed. My observations agree with those of the .Austrian ICxperiment 
Station that flacherie is not infectious. 

\nil. The true cause of the disease is the increase of certain products 
formetl In' the undue and rapid multiplication of warious microbes. These 
]:)roducts are in all probability no toxins, but the\' may consist of ammonia 
formed by jirotein decomposition, or of nitrite ftirmed from nitrate con- 
tained in the leaves, or of acids protluced from carbohydrates. \'ery prob- 
abU' these noxious substances are sometimes acting together. I hope to 
settle this {UK"sti(Mi satisfactoriK' In' further inxestiijations. 



44^ S. Sawjimura. 

The author must express here his sincere thanks to Prof. Sasaki who 
kindly transkited Ttah'an articles for him, further to Prof. Locz^' and Prof 
A'o.za/, and to J/r. Honda and Mr. llayaslii. Experts of Tokio Scricultural 
Institute who furnished him the larxx- and et^gs of the silk-worms, and finally 
\.v> Mr. Yavmsaki, .Assistant of tin- College. 



Zur Physiologie des Bacillus pyocyaneus, I!. 



VON 



O. Loe-w unci Y. Kozai. 



In Fortsctzuny" unscrcr friilicrcr Vcrsuchc,' cine mogliclist i^iiiistigc 
Xiihrstofflocsung fiu' den Jhic pyocyaneus z.u findcn, in wclchcr trotz Icbhaftcr 
Vegetation kcine Scbleinibildung aber reichliche Enzynibildung stattliabe, 
fanden wir folgende Lcesung diesen l^edingungen entsi)rechend : 



Tepton. o. 






Glycerin. o. i ,, 

MagnesiunisLilfat. o.oi ,, 

Dikaliuniphosphat. o. i ,, 

Natriumbicarbonat. o. i ,, 

Chlornatriuni. 0.4 ,, 

Das IMagnesiunisulfat wurde stcrilisirt bei der Infection zugesetzt. \\ ir 
\'a\"iirten in dieser Loesung die einzelnen liestandteile niehrfach und jedcs- 
nial war das Resultat cntweder eine langsamere X'egctation oder cine 
Verzogerung der W'iedcrauflccsung der Bactcricnmassen.- In dieser 
Loesung lauft die X'egctation in 18 — 20 Tagen bei 25— 28^C. ab. wenn die 
Kolben /////" rsur lliilftc V(j11 sind was behufs reiclilicher I-".nzyniprotluction 
notig ist untl jeden Tag kraftig vnigescluittelt w ird, wobei unter SauerstoH- 
absorption die gelbe Loesung tief griin wird. ICst \"oni 13. Tagc ab liort die 
Reduction des griincn I'arbstoffs auf. Die anfanglich reiclilichen l^actericn- 
niassen lc')sen sich ])is auf einen geringen Hodensatz allnudig w iedcr auf. 

> Sichc dicsc Lullctin,'-, Inl. 4. \o. 4 und X<\ 5. 

a Wir crliohtcn z. B. das Fopton auf ij?o', das Pikaliuniiiliosplial aut" 0.4%. wir cliiuinirlen das 
Nalriunil)icarl)onat und set/lcn cndlicli die Chlornatriummciigc auf 0.2 und 0.1 %' hcrah : audi 
vcrsuclitcn wir dieses dureh Natiiumsulfat zu ei-setzcn. 



450 0. Loov mid Y. Kozai. 

Diesc zuerst von EiiiuicritJi unci Laiv bocbachtttc Wicderaufloesung wurde 
von C onradi als cine Antol)'sc aufgcfasst, was aber wohl nicht dcni wirklichen 
Vorgangc Lnt>])ric]it ; dcnn Autolysc' ist die Gcsamnitheit dcr in einem 
Organ (odcr Organismus) nacli dcni Todc stattfindcnden fernicntativen 
Vorgllngc, cs wircl also als c'laractcristisch angcschcn, dass irgcnd cine 
vorlicrigc Sccci-nining von liiirjjnn nicht stattfindct. Andcrnfalls ist dcr 
Vorgang cbcn Icdiglich cine gcwohnlichc X'crdauung ; dcnn cs ist d(jch z. B- 
ganz und gar irrclcxant, ab cin Magcnsccrct den Magen verdaut. dcr cs 
abgesondert hat, oder einen anderen Alagen. Bei dcr W'iedcrauflocsung dcr 
gcwachscnen Bactcrienmassen durcli die seccrnirte Pyocyanasc muss erst 
cine gcwisse Anhaufung dcr letzteren in dcr Culturfliissigkcit erreicht wordcn 
scin. Dann erst kann dcr .\ngrilT auf die Xucleoproteidc- dcr Bactcrien- 
leiber luToIg habcn wwOi wird dann audi das wcitcre W'achstuni einge- 
schrankt inul endlich ganz verhindcrt." W'enn aber die l^acillcn auf festcm 
Xahrboden : Tilyccrin-Agar) cultivirt werdcn, so bleibt jedenfalls das J^nzyni 
in den Zcllen, wenigstens grossentcils. Dafiir spricht die Beobachtung von 
Krcvisc,"^ dass der Prcsssaft des B. pyocyanciis milzbrandhcilcnd wirkt/' 
Die Bacilien warcn auf Agarplatten cultivirt und die Vegetation nach 48 
Stunden nn't I'latinspatcl abgcnoninicn wordcn. Xebenbei bcnicrkt muss 
dicscr Prcsssaft ]<aum Toxin und ;>uch nur wenig Pyocyanolysin enthalt'. n 
liabcn ; dcnn 3 cc. warcii nach Krausc einem Kaninehen nicht schiidlich. 

Bei di m InteMX^se, welches sich an tlie P)"ocyanase knupft, suchtcn 

' Thcoliahi Smith liul licrcits i. J. \'i,i)\ die VL'rdaucndc Wiikuiit,' in stLTilcn Clcwcl.iLii von 
TliicTcii h-jubadUct ; si^iUcr li:iI)Cii Salkoiusky, y.ico'ii, I\f,!^iitis-Lcv\\ dicsc Krschcimini^ wcitcr 
Vfifolgt. Besontlcrs intcrcssaiit siiid die I-icsultatc Comadi's. 

2 Kraxohoio {^Ihfincislers licilragc 1, 530) liat ryocyni'.cus-Zcllcii ni:t vcrdiiiiiiu-ii Xatioii cxtraliirt. 
mil lissigsiiurc die Lojsung gefallL und nadi dcin Keiiiigen das Xuclcojirutcid analysirt. Er laiul dariii : 
^' = 5273/0; \\^G.^\%\ X--l6.5o;)u; r = 2.ll%; S^l,o%. In ^k:y\ Menilnanen lar.d er 
C^46.2%; H-^6.7%; Xi:.8S. Diescr SlickstolfgehuU deutct auf einc cliitinaitige Substan.?, \vas 
audi von Einmr/iti;^ fCu- ilie JNIenibranen des Vmc. (luorestens li(|uefaciens vcrnuitet wird (Her. Chcni. 
' ■i^^>. oV. 702). 

^ Xadi Sit-^warl (('. ]!ala. jo, 573) weiilen die Xude<>iroteide der Kacleriin ai:eli von repsln 
verdaul — aber erst naelidem die l!aeil!en ge'.o'.et >ind, was )r(]ri falls auffallei d i^t. 

•» Centibl, f. liakt. j/, Xo. 14. 

'» 'I'ypluiskunnte beini Meerseliweineben daniil niebl gelieill wenlen. 



Ziir riivsi(»l(»:;ic (Ir> I{ii('illii> |».voc>iiMcii>, II. 45' 

wir iKicli (.iiKr Mi^lliotlc. \wlciic bci L;i'<)sser I'j'iifachlKit tlocli cin rcincres 
I'roduct lirfcrt, als bisher ni()_L;licii war. UiisiT Ziel ist nuch niclit crrciclit 
wordcn, (loch ni(')<4en iinnicrbiii i.inii^c l^cobaclituiiL^^cn tier Mittheiluni; 
wci't scin. \'or cini^cn Jahi'LU Iiat /'"■ Xciuiibc'i^i^^ ubcr crfoli^rciclie P)C- 
haiKlluiiL,^ dcr Stapb)-l()ni)'k(jsis niit dcr P)'oC)'anasc i^RcjlifcrniciitUjcsuni^j 
bcn'cbtet. Dcrsclbc stclltc, w ic K. I'tn/'s/'- in scincn eiTob^rciclicn \'(jr- 
SLichcii dcr IMitzljrandbehandluiii;" niit l'}'oC)'anasc, dicsclbc in etwas 
\crscbicdcncn W'cisc dar, wic limincricJL und Locii\ nandich durcli Aus- 
salzcn nach ICrln'tzeii auf 58 (6 Stundcn;. Auf i L. dcr scchswochcntlichcn 
liouilloncultur \\urdcn 500 l;. .\mnionsulfat gcgcben,"' nacli 24 Stundcn das 
Ausijcschicdcnc cincr niclirtai^is^cn Dialysc iibcrlasscn und dann die Locsuni^ 
ini Vacuum zur Trocknc t;cbracht. Die alkah'sclic Locsuni;" wurdc also 
niclit erst neutral isirt, und in tier Tat liaben uns \erLjleichende X'crsucbe 
L,^ezeigt, dass dieses vcjrzuzielun ist. Heini Versetzen niit KssigsLlure' wird 
Kohlensaure frei, welclic 1)eini nacbfolL;enden Aussalzen in lilaseii festi^e- 
balten wird, so dass cine sebr scbauniige ^^lasse erlialten wird, welche 
schweer weiter zu behandeln ist. X c ucnb erg sow o\\\ wic Vacrst vcrwcndctcn 
liouillonculturcn. Dicse liefern cd^er cine sebr scbleimiL^c FIussit:jkcit,'' 
welche beini Aussalzen auch den Scbleim ausscheidet, tier nun einen 
L^rosscren oder i^erini^eren Theil der Tyocj-anase niit sicli reisst. W irtl nun 
dicse Ausscheitlunj; der Diah'sc unterworfen, um tlas Aniiiioiisulfat zu 
entfernen, so beiiierkt man eine aulTallentle Abnabme tics Scbleinis, sti dass 
man zur X'crniutunL;' kommt, es babe cin mit ausgcschietlcncs 1miz}"di 
(wcgen nun t;russcrcr Concentratit)n) tleii Scbleim durch JI\-drol}'se in niclit 
scbleimiije Trotluctc \'erwantlelt. Wenn tlie tliaK'sirte Locsuny," dann im 
Vacuum eini;etlam[)rt wirtl, so wirkt kein Scbleim niebr stt^irciitl, beim 
Locsen, resp. Injiciren ties l'rt)ducts. Wir liabcii aus i Liter Houilloncultur 

1 lIal)ilitation^^chl■il't, 15cni 1900. 

2 Ccntrbl. f. lJ;ikt. j/, No. 7. 

p Eiiic niassigc Vcrmchruni,' dc? S.ilzi'S luiiii^'l mir iiocli cine gciiiii;c ^k■h^.lu^■~cln.klulll; /u 
Wc-c. 

■* Es ist nalif zu 1 pioiiiillc Essiijsauiv bchuts Neutralisation notig. 

Diesc Sclilcinil)iklunt; beruht vicllcicht aut" Jcr Gegcnwarl niilclisaurcr Sal/e. Audi cssigsaure 
Sal/c uiul Asparagin licfcu schicimigc Culturen, IVpton aber nicht. 



45- ^>' liO'" iiikI V. Ko/iii. 

iiur 1.5 l;-. clcs Kuhfcrnicnts erluiltcn. I'rof. \itta bcobaclitLtc, iiach 
l-)anvicluini;" von 0.1 14". dcssclbcn /cy- cs, bci eincm Alccrschwciiiclien kcine 
Spur cine Tcnipcraturcrhohung; odcr ii'y;ciKl wclchcn andcrn Effect, cs 
warcn also kcine Substanzcn vorhandcn, die /cv cs hattcn schadlicli wirken 
konnen, was von einigeni Interessc sein mag", falls einnial dieses Rohfernient 
■/AW ]-)ek;'inii)fui\<4' von l^acillen (Cholera) ini Darm zur \'er\vendun_Li" kcminien 
sollte. 

W'ir haben nun die Aussalzniethode audi bei Cultiu'cn anl;■e^\•andt, 
welche ///V/// sc/ilcii/iig- werden, speciell bci der eini^anj^^s crwahnten Cultur- 
loesuni;'. Zehn Liter der 18 tai^it^en Cultur wurden zunilchst niit Chloroform 
versctzt und einen Tai;" stehen i;elassen, mil etwa noch \"c:»rhandene lebcnde 
Zcllen abzutOten. Am folL,''endcn Tage zeigte die Fliissigkeit cinen inten- 
sivcn (jcnich naclt hoidtril, es musste also ein i)rimares Amin in der Cultur 
L^eljildet worden sein. ]-)ie klare Loesuni;' wurde abgci^osscn, der letzte 
Theil filtrirt und in die Gcsammtmeiige der alkalisch reat^irendcn Fliissis^'- 
keit 10 \..j sechs Kilo Amnionsulfat eingetragen und unter haufigen 
Umriihren bci 6 — S" stchcn gclasscn. Es scliicd sicli nacli einiger Zeit cine 
llockige Masse an der Oberflrichc ab, welche abgenommen und tlurch 
I'iltration v\\\k\ I'resscn \'on der anluingenden ^Vmnionsulfatlocsung so gut 
wie moglich getreiint wurde. iJurcli dreitagigc Dial)'Sc wurde der Rest 
dcs Ammonsulfats cntfernt. Schon beini Anri'ihren niit W'asser wurde 
bemerkt. class sicli ein grosser Teil niclit wicder kiste, trotzdem wurde die 
(Icsammtmassc in den Dialysirschlauch ' gegeben. Der unl(')sliche Theil 
war \-on einer melaninartigcn Substanz schwarz gefiirbt, und cnthielt neben 
blaugruneii l')'oc)'aneusfarbstoff noch einen gcringen Anted lu>hercr Eett- 
siluren, und etwas I'roteinsubstanz. Das l-'iltrat wurde zunilchst auf 
vcrdaueude W'irkung gL-pruft, aber nur cine ilusscrst schwache W'irkung auf 
gcc[uollenes j-llutfibrin beobachtet, sclbst als noch 0.2^.'y Soda zugcsctzt 
wurde. Dar.ius durfte wohl der Schluss gczogen wertlcn, class die l*)'ocya- 
nasc kcine AlbunKJScnatiu" bcsitzt,- sonst wilrc sie niit ausgcsalzcn worden. 
Bci den Versuchcn von Ncucnbcrg und \oii Wicrst musste wohl die \'olumi- 

' /u .iiili.ii.'i>liclii.in /uLxki.' uiuilc .lucli (.■t\\as Cliloiul'unu /.u_L;uscl/.t. 
* W.-iIirscIieinlitli .iliiK.-lt >ic ticii iV'iitoncii. 



/iir IMiv>i(il(»ni(' dcs Biuilliis p.voc.v.iiions. 11. 



453 



iv'isc Sclileinimassc, (li\; ausL;'cs;iIzcii wurde, \'icl Mny.ym niit nicdcr^'crisscn 
habcn. 

l^s ist (Icssliall) wolil dcr Schluss q-crcchtfcrti^t, class hci iiicht sclilci- 
mii;"cn ("ultui'cn die Abdami)rmcth<)d(.- im Wacuum) dcr AussalznictlKxlc 
xor/iizii-lun ist, da sic siclur die ( icsamnitmcn^c dcs l'",nz)-iiis licfcrt. 



iJber den Kalkgehalt der Milchdriise. 



VON 



M. Toyonaga. 



Ich hal)c in mcincr fri'iliLren iXrbcit iibcr tlcn Kalkijchalt dcr i^a'aucn 
unci weisscn liirnsubstanz clarauf liinQ-cwicscn, class die Driiscn im X'crhLiltnis 
zur Mai^ncsia vici mchr Kalk cnthaltcn als aiulcrc Gcwcbc des Ticrkorpcrs, 
was jodcnfalls mit dcr Qrosscrcn Zcllkcrnmassc zusammcnhilnj^t. l\s war 
in dicscr l^czicliunL;" natiirlicli von Intercssc dicsc UntcrsuclninL;"cn fortzu- 
sctzcn, insbcsondcrc wcil in IVzug auf die verscliiedenen Ori^ane ties 
Ticrkorpcrs auffallcnd weni^'c Aschen-Anal)'scn vorliei^'cn. widirend in 
l^ezLic;' auf den Pflanzenkorpcr dicsc ausscrst zalrcich sind. 

Ich habe zuni'ichst die Arilchdri'ise in l^ctracht ^'ezo£;"en. welche ins- 
bcsondcrc deslialb l^cachtunq; \erdicnt, \\cil ihr Secret in l^czuq; ant 
Arincralbcstandthcile _e;anz anssemrdcntlicli \on dem Hlute diffcricrt ; so 
fa nd Biiiio-c: — 

loo Tlicile Asdic viitliailcii : 1 luii'lciiiilcli. 1 lumk-liliit. 

K„() 10,7 3.1 

Na^O 6,1 45,6 

Ca O 34,4 0.9 

Mo-O 1.5 0.4 

FC3O.5 0,14 9.4 

1'.. <^r, 37'5 13.3 

CI 13,4 35'6 

W'ir crsilicn hitraus in Hezui;" auf den Kalkt;'ehalt _q;anz enornn^ I'ntcr- 
scliiede. In der 1 bnukMnilch bercchnct sich ilas \'crludtnis 

Mo-0:CaO=i : 22.93 
in liundibhilc I\TqO : CaO= I : 2.2; 



45*^ ^t- Toyoiias'ii. 

Tell bescliraiikti.: mich Ix'i dcr Analyse auf die Bcstimmunq- des Kalks 
and del" AIac;ncsia, da besonders tlieses Verhaltiiis fur die \ersehiedenen 
Gcwebc sehr charakteristiscli, und die A?che der ^FilclKlnise iiberhaupt nnch 
iiiclit untersucht ist. 

Ich trennte bei der Milelidriise einer Kuli so ^ut als m()glich das 
]^indcc(e\vebc von der cit;er.tlichen Driisensubstanz ab und bestimmte y.u- 
niiclist den Wassergelialt, derselbe betrut^" 66.7^'Q. 

Nun wurden 8(S,r)4T c;" Trockensubstanz mit 5 c;" wasserfreiem Xatrium- 
carbonat i;emischt und verascht wobei das ^^"eissbrcnnen wie gcwrjhnlicli 
schr lan<4"e daucrte. Die Masse wurde zunilchst mit Wasser extrahiert und 
nach l^ntfernunq" des kohlensauren- und jihospliorsauren Natrons der 
ausc^rcwaschene Rlickstand mit Salzsilure i^x-lost, wobei eine Minimalmencie 
Kieselsiiure unq;el(')st blieb hiirauf die L(')sun_g; mit Ammoniak bis zu 
alkalischer Keaktion versetzt und dann mit l']ssi<4"s;uire bis zu schwach 
saurer Iveaktion vermischt. 

Ilierbei bleibt ein geringer flockio-cr Niederschlai^" von phosphorsaurem 
Kiscn uni^el()st. Aus dem Filtrat wurde nun der Kalk mit oxalsaurem 
Ammoniak j^cfiillt und das eini^cenii^te l-'iltrat \-om Kalknieck-rschkiL;" zur 
]\raf;nesiab(.stimmung vcrw. ndet. 

ICs wurde erlialten : 

CaCO^ —0,39951,^ =0,22311; CaO 

^r^^^^'oO, = o, 1 562 o- = 0,0566 o- Mo-O 

lli(.•l•au^ licrcclinct sicli •■- •, , 1- , , , 

lui- 1000 ieilc fnsclicr Druse : 

CaO =0,8401 Teilc 0,2517 Teile 

Ml>0=:0,2I31 ,, 0,0639 ,, 

W'r_L,deiclien wir das sicli hieraus erqtbendt; \^erli;iltiiis ,' mit ilen fiir 
Milz und Niere von Alo>' ' L^'efundenen Zahlen und mit dm ZaliU'ii fiir d.is 
Mu^kelfieiscli von SauL;etii'ren, so crcjiebt sicli : 

MikliilrU-c. .Milz. I'ankrcis. Nicir. SiiugL-tior-Mu-krl. 

-^~ 4,67 6,79 4,05 i,S4 0.34 

]'",s ist somit audi bii der MiKlidiiisi' wir bei der andcrcn I M'iistii der 



' |.iliic~liriit lit 1. 'riiicicluinic 30, S. /|<)J. 



1 



IIh'V ilvn U'alkgciiiiK dcr MilclidniM- 



457 



CalciimiL;xli.ilL !.;r<'>sser .ils dcr l\I;iL;ne.siiiinm.li,ilt, witluxiKl fiir das Muskel- 
i;c\vcl)c (ler \\ 'arniblutcr uniJ^ckclirt ckr ^tlai^iicsium c^elialt L;rosser ist als 
(Ut Calciunigclialt. 

\'cr<;lciclicn wir iUKdi die Aren_L;\n xon Ca und M;^^ im MusKl-I ivn't dciK ii 
in Milclidriisc und Mil/, so hat man fur die organisclie Trcjckensuljstan/ : 

Saugelliicr yi///j/{v/(K'a(z). MihJiJruse. Ali/z (Ribaut) 

Ca o.033;"'o^ O-I73/0 0.141% 

ATg 0.109,, 0.038,, 0.056,, 

l""s ei'f_;ibt sicli soniit, dass nicht nur der C alciuniL;elialt absolut i^a'dsser ist 
in der Driise wie ini ?*Tiisl<eI, sondern aiicli dass der Ma^'nesiuni-_,a:]ialt dort 
welt uei'inLier ist als liier. Ich werde nieine Untei'sucluinu'en fortsetzen. 



Der Erntequotient. 

Oscar liOew. 



Tu alien vullstruKlii^en lu'ntcbcrichtcn aus dcr Praxis sowohl, wic den \'cr- 
suclis-Stationcn, wircl ausscr dcni wcscntlichcn iM'ntcbcstandtcil, wic Knollcn, 
Wurzcln, Friichtcn audi nDcli die ]\Icn[;c dcs Krautcs (nlcr Strobes angege- 
ben. Aus diesen Zablen ersiebt man aber nicbt sofort, ob sicli das Ver- 
haltniss zwiscben diesen iK'standteilen deni Mittel oder eineni Optimum 
nidiert. Beluifs einer sofortigen Reurteiknig dieses Verbiiltnisses moclite 
icli den Ik-griff des EnitcquoticiiUii einzufi'ibren \'orscblagcn. Ya gostattet 
sofort zu erselien, <^b unter den gegebem-n IV'dingungcn (Bodcn, Dungung, 
Wetter, etc.) ein mittleres oder o[)timales VerblUtniss crziclt wurdc. Kr 
gibt die Hauptleistung der l^latter, der wicbtigsten I'roihicenten organisclicr 
IMaterie, in vergleicbbaren Zaien an, er zeigt, ob diesc Organc ibrc Aufgabc 

\()1] und ganz erfullt baljen. Dicser ba-nte(iuotient 

k 
q — . lOO 

s 

tb'iickt die iM-nte ties wesentliebsten liestandteils k ^ Korner, Knollcn, Wur- 
zeln, in I'rocenteii der lilattsubstanz, dcs Stroll's, s. aus. Abui kann fin- die 
Zwcckc tier I'raxis tlie ("icwiclite des lufttrocknen Krautcs oder Stre^bs zu 
(iriuidc legen, widu'eiid Rir rein wissenscbaftliclie Zwcckc this Gcwiclit dcr 
absoluten Trockensubstanz zu dii, iien lu'ittc I'.s wilre von cinigcni \*ortcil. 
den absoluten b^rntcwerten pro ba .uicli ilen I'a'ntcquoticntcn. iler sicb bci 
nornialcn l^tkmzcn oft zwiscben geiiau Ijcstimnitcn Grrmzen bewcgt, bcizu- 
fugen. So bctrrigt (,lerscll)e iiii Mittcl bci ( k rste 73, wabrcnd cr im Optimum, 
wic cs wobl in der Praxis nicbt errcicbt wird, lOO bctragcn kann.' IVi 
15ob nen licgt er in der Kegel weit iibcr lOO, b^i b'.rbscn baufig-iibcr 200. 

> IfLllru.;cl tcilt mit, d.iss cr uutcr sclir i^unsliycii IJciliiij^uni^ca iiu Glashaus bci Gcrste glcichc 
Ucwichtc Stroll uml Kiuiicr t;ccnitct Iial)C. .\iulcrcr.-;cits l>c;schrcibt E, /PV/y Fcldvci"suche, wclchc auf 

100 Stroll iiur 60 1 111. Krinici" ;_;ahcii luul lici Wci.cn i^.ir nur 40. 



4f3o Oscar lioov. 

I'Vi'iKT wiirtlc er. sicli iin Mittcl aus zalrcichcii Oaten crL;cl)en li'ir 

\\'L'izcn 53 

Hafli- 66 

Mais = . .. 80 

Scnf 52 

Hucliwcizcn 54 

WOlil habcn schon vcrschicdenc Forschcr den KornerertraL;' hie uml tl.i 
auf 100 Tlieile Stroh bezo^^en. al^er es ist systeniatisch wecler der niittlere 
noch der optiniale ,, l:rntcqiiotii'/it " bestininit worden. Besonders war es 
/'. Warner, welclier seine Rcsiiltiite niit Cerealien in dieser Form ausdruckte. 
So fand er z. B. dass bei verschieden starker Stickstofftliinguni^ auf lOO Till. 
Stroll resultireii konneii bei Hafer 5 1 87 Till. Korner, bei Rogs^eii 46-53. 
bei W'eizen 33-63. Ferner hat er bei Mafer auf lOO Thl. Stroh 52 Till. K(")r- 
ner erhalten, als er Cliilesalpeter bei der I'j'nsaat t^ab, aber 64 1 hi. K(')riier. 
wenn er diesen bei bes^innendeni Schossen zufut^te.i 

Gewisse Verhaltnisse fiihren zu einem Ueberniass von Hlatt})roduction, 
anelere wieder ermoyliclien den J-Jlattern, die \o\\ ilinen bereiteten ori^ani- 
schen Xahrstoffe in ausi^iebii^ster W'eise der AusbilduuL;' der I-'riichte zu- 
komnien zu lasseii. Diese Arbeit niit einer Zal auszudriicken, beabsiehtii^t 
der Rnitcqiioticnt. 

' I lie Slick>lM|'t.luiii;uii;4 iLt l;mil\\irUi.liaUliclien (_'ulturi)ll.ui/.ai, ibiyj, S. 164. 



Ueber die physiologische Wirkiing des ChlorrubidiLims 
auf Phanerogamen. 



V( )\ 



Oscar Loe-w. 



Vcrsuclic mit lUichwcizcn batten mir fri'ihcr c;"czciL!,"t. ' class cine i)liysio- 
loMisclic X'crtrctun^' vnn Kalium clurch das ihm so nahc stclicntlc Rubidium 
iiiclit m')^]icb ist Zu (h'csuni Scbhissc zwar scbon vor mir Birncr und 
Lucajius- q'ckommcn, allcin icb constatirtc immcrbin ciiicn i^Tossen Unter- 
scbicd zwiscben dcr W'irkunc;" \'on Rul:)icHumn!trat un.d Ivubidiumclilorid. 
Alit Xitrat crt^abcn sicli patliologiscbe Starkcanscboppungcn, cine \'cr- 
cHckun<4' und Torsion dcs Stcnt^cls, Sistirunf^ dcs Liingcnwaclistums, Ivimoll- 
cn iukI Fk'iscbiL;\vcrdcn dcr IViiittcr inid scbbcssbcb crfolgtc dcr Tod, bc\"or 
cine l^liitc cntwickelt war. W urde Ljlcicbzcitii;' ein Cblorid (Sabniakl zu- 
o-csetzt oder RubicHum niclit als Xitrat. sondern als Cliloritl \"cr\vcndet. so 
strcckten sicb die rHanzen und t^ckanf^ten nacb l^rrcicluuiL;" ciner wcit beilcu- 
tendcrcn Ibibe bis zur J-)Kitcnl)ikhm<4" was deutHcb fiir (\c\\ k-influss \"on 
Cbloriden auf ^\<^\\ Stilrkctransport spriclit. Mrst nacb ck-r l^Kitcnbikbuii;" 
tratcn 1 lemmungserscbcinungcn ein. cs kind cine Anbiiufung \-on Zuckcr uiul 
Vcriiiulcrungcn dcs Cbk^roplndls statt unil tUe klkmzcn \"crhelen cincm k-ing- 
samcn Siccbtum, obnc eincn Samen i)roducirt zu babcn. W'citer gckingtcn 
Pflanzcn, dcnen Kabum und Rul)idium zugleicb gcgcbcn \vur(k\ indem die 
1 kdfte des in ck-r Controllocsung \"er\\enck-tcn Cddorkabums durcb Cbk~»rrubi- 
tb'um ersetzt war. Indesscn aucb bier wurck' die 1 b">bc dcr Controlpflanzcn 
nicbt errcicbt y\\\(\ kc-in reifer Same gcbikk-t. (.be Ttkinzcn starbcn nacb ikr 
l^b'itcnpcricKle al). 'i'rotz dcr patlK^k^gisclicn W'irkungen ergab sicb immcrliin 

1 l.amlw. Wis. Stat. 21, S. 3^:9. 
■-• ll'iil. 7. S. 26 ^ 



4^)2 Oscar Loow. 

fur (las Rubidiuni cin physiologischcr Nutzen, den das Natrium nicht bcsass, 
dcnn die Pflanzcn producirten weit mchr Trockciisubstanz, was vicllciclit 
nur auf cincr Untcrstiitzunc;" dor W'irlcung dcr im Samcn gcspcichertcii Ka- 
liumsalzc bcruhcn maq-. Von Interessc ist liier die Beobachtunc;^ von 
jMoliscJi,^ dass Algen in ciner Culturloesung sich gar nicht cntwickeln, wenn 
darin statt der Kaliumsalze Rubidiumsalze vorhanden sind. Ist es hier die 
Zellteilung oder die Assimilation des Kohlenstoffs, oder die Eiweissbildung 
oder sind es diese drei wichtigsten Vorgiinge zusammen. welclie mit Rubidium 
statt des Kaliums niclit ausgefiihrt werden konncn ? Diese Frage konnten 
\-ielleicht Vcrsuche mit Pilzcn entscheiden. Hier beobachtctc ich nun. dass 
l^ierliefe und der gemeine Pinselschimmel sich sogar noch besser entwickeln 
k(')nnen, wenn bei Zucker als organischer Kohlenstoffquelle Rubidium statt 
des Kah'ums dargeboten wird. Beide Klemente kamen als Tartrate zur 
Verwendung. \\' erden jedoch wcniger gute Kohlenstoffqucllen, wie Xatrium- 
acctat, verwendet, so stosst man auf einen bedeutenden Untcrschied zu 
Guiisten des Kaliums. Da nun verschiedene Pilzc cine verschiedenc W'achs- 
tumsgeschw indigkeit besitzen, somit \\ahrscheinlich der iMweissbildungs- 
process mit ungleicher ]'\'rtigkeit ausgefiihrt werden diirfte, Hess sich ver- 
nniten, dass die Verwendbarkeit des Rubidiums bei diesem Process audi 
nicht stets mit derselben Leichtigkeit vor sich gienge. In der Tat hat (u'ui- 
ther- beobachtct, dass wiihrcnd der Pilz Botrytis ciiicrca Rubidiumsalze 
physiologiscli \crwertcn kann, cHlscs bei RJiir:.opus nigricaiis nicht der 
VaW ist. Ich beobachtctc cine Wrtrctbarkeit bei jHicicriinii coU und. 
wcnn audi in weit geringcrem Grade, bei B. pyocyaucus, widircnd bei 
Cloih'thrix or/on'fiiui sell)st bei Zucker als Xiihrstoff cine \'ertrttung sich 
als unm(')glich crwics. 

Kubidimnsalze zeigen somit in physiologischcr Beziehung cin eigenar- 
tigcs V'trhaltcn. Die oben erwahnten pathologischen PLffecte beim Buch- 
weizen einerseits, die giinstigen l^ffecte bei llefe und Schimmcl andererseits 
veranlassten midi, die N'ersudie mit Phaneroganicn in modificirter Form 
wifdcr aufzundinicn. Ich \crsucht(.' die W irkun;.; klcint r J )(iscn Knbidium- 
chlorifls i)(i rikm/.t n nntcr nurmalcn l''.nKdniingsl)c(h'ngungcn. 

> Wicii. Aka.l. l!cT. 1^96. 

* Iiiauguraldissciiation, Kilaiii^'en iSoj. 



Ueber die physiologischc Wirkiing dos Chlorriilndiiiiiis iuif Pliaiiorogriimen. 463 



Vcrsucli init Brnssica chiiiensis. 



Drei Tr)pfc mit je i Kg-. Bodcii wurdcn gediiiic^t mit : i 1;. Kaliumnitrat, 
05 £^ Ammonsulfat, und 0.5 g. Monokaliumj)hos[)liat. Ausscrdcm crhiclt 
Topf a, 10 Milli^^ranim Rubidiumchlorid, 
„ b, 50 

,, c. dientc zur Controllc. 
Der Chlorgehalt des Bodens entsprach nahe zu 0.05 £,''. Na CI per Kilo, er 
war ein Ichmiger Boden, zum Teil aus vulkanischcr Asche bestchcnd. 

Am 21, October wurden 10 Samen pro Topf ausgesi'it und am 5. Xo- 
vcmbcr die junq-en Pflanzen auf je drei moglichst t^leich q-rosse, 6-y cm. hohe, 
rcducirt. Gegen Mitte November ergab .sicb, mit Ausnahme einer Pflanze 
in b. fiir die Rubidiumplianzen ein besseres Wachstum al? fur die Control- 
pflanzen, ein Unterschied, der mit der weiteren Entwicklung immer bedeu- 
tender wurde. Am 17. December ergaben die INIessungen fiir das Uingste 
Blatt jeder Pflanze Folg-endes : — 



c. Controlpllanzen. 



21.0 cm. 
22.4 ,, 
28.2 „ 



14.0 
19.1 

25-5 



16.5 
17.0 
21.1 



Am 22. Dec, wurden die Pflanzen ausgezogen, die Wurzeln gereiniqt 
und mit Pliesspapier gut abgetrocknet, und die ganzen Pflanzen ge\vo"-en 
im frischen Zustande, mit folgendem Ergebniss : — 



a 


b. 


c. 


14.3 g- 

16.7 „ 

1 8.8 „ 


6.1 
14.0 

25.2 


10. 1 
10.2 
13.0 


Mittel : 16.6 


15.1 


11.8 



464 



Oscar Loew. 



Es war somit ein stimulirender Effect des Rubidiumchlorids zweifellos, 
doch war dieser bci Erhohung von lo Milligramm auf 50 pro Kg. Boden 
nicht vermehrt wordcn, die Pflanzenmasse war im Gegenteil in letztrcm Falle 
etwas kleiner als im ersteren. 



Versiicli iiiit Gcrste. 

Zwei Topfe niit je i Kg. lufttrockncm Bodcn erhielten als Grunddiing- 
ung je 1.5 g. Ammoniumsulfat, 0.5 g. Monokaliumphosphat, 1.5 g- Calcium- 
superphosphat, i.o g. Kaliumcarbonat ' und 0.5 g. Natriumnitrat. Einer erhielt 
atisserdem noch 0.2 g. Rubidiumchlorid, der andere die aequivalente Menge 
Natriumchlorid. In jeden Topf wurdcn am 14 October 10 vorher gequollene 
Samcn ausgesiit und die Entwickkmg im Glashause wie beim vorigen Ver- 
such beobachtcd. Am 21. October wurdcn die Pflanzen auf 4 pro Topf re- 
ducirt, so dass allc von m()glichst der glcichcn }I('")he waren. Gegen Endc 
November zeigtc sich cin deutlicher Kohen-Unterschied zu Gunsten der 
Rubidiumpflanzen, der stcts zunalim. Die Messung am 17. December 
ergab : — 



Rb- Pflanzen. 


Control- Pflanzen. 


44.6 cm. 


39.1 cm. 




4^^- 5 .. 


46.0 ,, 




4<''..S .. 


47-0 M 




54-3 w 


4'>5 .. 





\)\c Ii()licn-Untcrschicdc nahmcn zu, wic die am 19. Januar anfgcnom- 
mcne photographic (Tafcl XXV) gut crkennen liisst. Dabei waren die Rubi- 
diumpflanzen vollstiindig normal.- Wegcn Auftretcns von Pilzcn wurdcn die 
I'rtan/.cn schon bald nach der l^)lutenpcriodc gcschnitten. Das Gcwicht 
betrug : 

* Das Kaliunicarhoiiat wiinio siiiiler scparat dfin llodcn cinvcrlcil)t 

2 ()1) Hud)\vfi/c-ii iiiid aiuliTc I'llaii/i'ii lik'la'i ihi-iit'all- normal lilrilicii, m)11 nocli ijipriift weiilen. 



lifbci" (lie phv^iolo^^isclic ^ViiKtmi: <!(•■> ( Iiluiriiltidiiiiris auf I'll iiici"o:.';iiiii'ii. 4^>S 

l\ulji(Iiuiii[)llau/L-ii. C'liitroljjd.mzcii. 

.Aclircn, I'l'isclii^cwicht ; 6.1 i^. 3-7 k^- 

Lcbciulc I')l;lttcr, frisch ; 81.3 ,, ^3-3 . • 

Abt^cstorbcnc l^lilttcr, luftti'ockcn ; 5.2 ,, 4.8 „ 

I'>in drittcr Wrsuch wurdc niit Spiiicaca olcracca aiiLjcstellt. AIlc V'cr- 
haltnissc warcn liicr die L'lcichcn wic (jben bci J>rassu(i. IVi deii Ixiiljidiuni- 
pflanzcn crhielt der I'olIch 50 Milligranini Kiibidiumchlorid per Kilo. Als 
dcr Sanicn rcif war, wurdc gcschnittcii iind die bcstcn lvxcnii)larc friscli 
i^cwui^cn. 

KuhidiuiiipllaiiZv'u. ContioliJllanzcii. 

(jc^\icht der Lj;r6ssten Pllanze, \'arietat I 18.2 lj. 12.O i^. 

Die zwei t^rossten Pllanzen der X'arietilt II ... 16-5 i.,^ 13.2 l,^ 

Es hatte soniit in alien diesen Fallen ein stimulircnder Effect des Ivubi- 
diumchlorids stattgcfunden, was wohl \on betrachtlicheni tlieoretischen In- 
tcrcsse ist. Fiir die Zwcckc dcr Praxis jedoch ist cine Anwendun:^^ des Salzes 
ausgcschlossen, da dcssen Prcis ein zu hoher ist.' 

1 V.i koicn 100 i;. Kl) CI ;:: i; Marl< (ui. 6 Vcu). 



(Jn the Stimulating Action of Manganese upon Rice. 



BY 



M. Nagaoka. 



In our hist JJuJlctiu the observation was ccjniniunicated 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 carr)- on a 
field exi)eriment with rice which is the most important a<^ricultural plant in 
Japan. 

Thirty six wooden frames each representing^' an area of 0.826 square 
Meter were placed, tlnx-e feet apart, into the paddy field of our CollcL^e farm 
to a de[)th of 60 cm., leavini;- 6 cm. above the i^round. The soil had not re- 
ceived ail)- manure the ])revious three years" and was now manured' in the 
ratio 

c^f 100 Kl,^ X per ha, as annnonium sulphate 
,, ,, KoO ., ,. as potassiimi carbonate 
,, ,, 1*2^^5 M .. ii^ double superjjhosphate. 

The potassium carbonate was seiKiratel\- a])plied (June 23) and the 
other two salts four da)-s later. 

On June 2*) manganese was api)lied as manganosulphate in such quanti- 
ties that the amount of manganic oxiil corresponded to the followint:^ propor- 
tions, tliree series being obser\ed in each case : — 



1 Kcloic inaimriin:; llic soil was silted, and all rciunants ol" former vegetation removed. 



468 



M. >'ji8-aoka. 



No of wooden frames. 



Mn, O^ per lia 
Kg- 



Gin 111. 



I 


13 


25 








2 


14 


26 








3 


'5 


27 


10 


0.S33 


4 


i6 


28 


15 


1.250 


5 


>7 


2y 


20 


1.666 


6 


iS 


30 


25 


2.0S3 


7 


'9 


31 


30 


2.499 


8 


::o 


32 


35 


2.916 


') 


_'i 


33 


40 


3-332 


1(1 


-- 


34 


45 


3-749 


II 


^3 


35 


50 


4.165 


1 z 


-4 


36 


55 


4.582 



On Jul)- 7 the youni;- rice pkmts ^55 days old) from the seedbed, were 
transplanted into the frames, each receiving 16 bundles of twelve healthy 
indixiduals of ecpial size.' The treatment (irrigaton, etc.) did not differ, 
fr(jm that usually observed with tlie rice fields in Japaii. The weather con- 
tlitions were not favorable this year for this crop in the whole haiipirc of 
Japan, but the relatixely low sununer temperature dinn'nished on the other 
liand the dangers from fungi and insect pests with this crop. Our frames re- 
mained free from such pests. The croj) was harvested on November 29 
with the following result, obtained by weighing in the air day condition. 



' Tlie variety was the Sii/.ui///,t, cliaiacleiized \>y its resistance j'ower and -.nediiim duration of 
vegetation. 



On llio S4i ill 1 1 lilting- Aclioii of Miniiraupso upon IJico. 



469 



No. of 


l-)cr ha. 


Fu!l 

grain'?. 

gr. 


Enipt y 

grains. 

gr. 


Stra-.v. 

i 




AVERAGE. 




frames. 


Full 
grains. 


Empty 

' grains. 

1 


Straw. 


Total. 


I 
13 


no manure 
and no 


151.6 
142.6 


.1--' 
30 


! 
193.0 

1 71.0 


150.3 


1 

3-0 


1S50 


33S.3 



25 I Mn, O3 i 156.5 



2 no Mn, O3' 1770 



2.7 



191.0 



14 

26 



3 
IS 
27 



4 
16 
28 



5 

17 
29 



7 

•9 
31 

8 
20 
32 



25 



227.S 
202.6 



4.6 
6.3 

5-2 



242.0 I 

3I2.S 202.5 

254.0 1 



250.6 ! 6.8 
239.6 5.S 



251.S 



8.4 



319-6 

285.6 
:;2i.o 



I 249.9 6.3 401.6 

15 I 257.6 4.8 307.5 

I 262.6 : 5.1 278.0 



!56.7 



277.5 ! 5-3 I 354.6 
269.4 1 7.8 341.6 , 264.3 ! 

245.6 , 5.9 i 287.0 ; 



279.0 j 


10.3 


330-^ 


264-5 


5-1 


335-0 


272.7 


5-2 


326.0 


270.0 ' 


5.8 


374-7 


256.5 ! 


4-4 


316.0 


276.S 1 


5-3 


332-0 


264.6 ; 


8.2 


325.S 


267.S ! 


4.4 


3340 


269.4 


5-5 


309.0 



267.7 



267.3 



5-4 



269.6 477- 



247.3 7-0 '■ 308.7 564-0 



44 



320.1 



6-3 



327-7 1 59S.3 



6.8 



34S.5 ! 627.4 



3.40.0 61 2.8 



6.0 



322.9 



596.2 



470 



M. Xascaoku. 



No. of ^^"^^^3 



frames. 



Full , Empty St:;\\v. 

per lia. ' grains. grains. 

l<g. nr. gr. gr. 



a\era(;k. 



Full Empty 

trrains. ijraii.s. 



Straw. : Total. 

i 



40 



261.8 ' 8.0 

269.3 5.6 

285.9 ' 7.0 



364.6 
30S.0 
343-0 



6.9 



338-5 i 617.7 



45 



34 



256-5 
2S6.5 
272.8 



9.0 I 340.6 

] 

7.1 i 33S.0 
5-3 i 326.0 



271.9 



3J4-9 i 613.9 



-.1 

35 





? (iq8.6) 

1 


(9-0) 


312.5 










50 


270.6 

! 287.6 


6.7 
6.5 


399-0 
367.0 


278.. 


6.6 


359-5 


645.2 


1 


1 

254-5 


6-7 


331-4 










55 


279-4 
283.9 


5-2 

6.4 


364.1 
340.0 


272.6 


4.1 


.345-2 

1 


621.9 



The application of maiiL^ancsL' lia<l therefore a considerable influence 

upon the yield, which will be noticed more convenient!)' by the followini;- 

tabic, in which we take the yield in i^rains of the manured plot without 

man_L;'anese as a luiit :- 

Mn.jO^, Ko-. ])er ha. Harvest of full i4ralns. 

(Average) 

none i-OO 

15 1-26 

20 1-30 

25 1-34 

T^o 1-32 



35 
40 

45 
50 
55 



1-34 
'•34 
'•37 
1-3-I 



Oil tilt' Stiuiulatiii;L;° A<-1ioii orMaii^aiiOKC ii|»(m Uico. 



4/1 



It will be noticed frrmi these: figures, tliat a moderate dose of 25 K'\\n 
MnoO., l)er ha led to an increast; of the liarvcst of one third and tli.at 
hig-hcr doses of Mn2<):; did not influence esscntiali)' tin's result undir the 
_C;iven conditions. 

It is furtlier of sonie interest to examine whether the axera^c ratio 
between tlie \\eiL(ht of i^rain and straw is affectcti tf) an)' extent by the in- 

n r 'iM ■ r-iii"^ (X) . 

nucnce ol manganese. 1 lie ([uotient ot )-ield' -r which expresses 

the i)ercentaL;"e of ^rain relative!),' to straw is for the different cases: — 

MAXURKD PLOTS. 



Aiiidunt ol .\lii,j< >3 i>tr li 



( luotient 111" VieliJ. 



No niangar.cse 



1 o K o 

"5 M 

20 ,, 

35 .. 

;^o „ 

?5 " 

40 „ 

45 » 

SO „ 

55 „ 



7S i 

7S 
S2 
So 
Si 
77 
79/ 



I he application of man_L;anese' liad therefore — tY/rr/s /'diibiis — a fu or- 
al ile inHuence on the quotient of )-ield. 

Let us now determine b)' calculation w hether the a]")plication ofman^ani-*- 
sulfate would be profitable for the farmer. The i)rice of \oo \\.\\o of pure 
cr)-stallized manq;anosulphate is according- to the latest pricelist of Tliec^dor 
Schuchardt= iio ^NFark or 53 \-en.2 

The average production /tv /ta, of <^rains of rice with husk is= ^-r^-^-r 
Ki/o and of aiia1r\' straw = 5250 Kilo. 



1 ( )n the Qiioliciit of 3"/V.'</' ^^luiUcquotionl) soo tlu- artMo ol" O. /..'fw in \\\'\< liulli-tin. 
' I Mai k - 3S/„8, ?cii, latest quotation. 



4/2 M. >iiyaoka. 

Tlic wholesale price of. crude rice £^rains is 9,9 sen per Ki;., of airclry 
straw = 1.2 sen. per Kq;. hence tlie a\'erat;"c yield per ha. has a value of 349 
yen in grains and 63 }'en in straw = 41 2 yen. 

.An increase of one third would ha\-e an additional \"alue = 1 37.33 yen 
while the cost of the many;ano-suIphate recjuirctl would be = 30 yen. 
Hence the application of this salt on soils poor in niangancse would be of 
advanta£,a". 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 Iodine and Fluorine 
Compounds on Agricultural Plants. 



BY 



S. Suzuki and K. Aso. 



A. On the Influence of Potassnini lodid on Oats. 
By S. Sitzuki. 



I have demonstrated in a former article', that potassium iodid in 
exceedingly high dilution can exert a stimulant action on plant growth. 
The pea had served for that experiment. I had, however, at the same time 
commenced an experiment with oats, the result of which arc 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 NO3, 3g.K2CO.; 
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 o.oig. potassium iodid dissolved in 100 c.c. water ; further 
pot No. II. o.ooig. and No. III. o-oooig. of that salt, while No. W . served 
as control. Those t[uantities of potassium iodid expressed in percentage 
of soil are : 

No. I. — 0.00 2609 % 

No. II. = 0.000 2609 % 

No. III. = 0.00002609 % 

Hull. College of ^V;^riculture, Toyko \'oI. 5. Xo. 2. p. 11^. 



474 



S. Suzuki and K, Aso. 



In the beginnint,^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 XXVIV The 
plants were irrigated almost daily witli 300 c.c water until the flowering 
stafTc was reached, after that with 500 c.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 : 



Xumber of Stalks, 

Weight of grain?, un- 
husked, g. 

Weight of straw, g. 



I. 



11. 



ITI. 



'5 

24.S 

48.5 



14 

-5-5 
56.6 



14 



5S.4 



21.4 
45.2 



The resul'i. undoubtedly proves a stimulant action of iodine, even if 
present in such a small ciuantity as 2. 6g KJ in 10000 Kilogram of soil as in 
No. 111. The increase however, is with oats not so large as with the pea 
(These ]3ulletins, vol. V p. 199). 

I had mentioned already in my former article, the experiment of 
A. Vu'/i-h-r vv'ho soaked seeds of wheat and barley for a short time in a i^^^," 
sohition of sodium iodid and observed with such seeds, an increase of yield. 
The quantity of sodium ioditl 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 \% potassium iodid solution, washed and then sown 
in two pots, 15 seeds in each. Later on the young shoots were reduced to 
five of eciual height. After a few weeks, it became clear that the plants did 
not so well develo]) as the control ])lants. This may be due to more iodid 
having entered into the grains than in the case described by Wrlekcr. 
This difference is probably caused bs" the prolonged soaking in my case. 
Also differences of temperature during the soaking process can influence the 
result. The i)lants were cut nu July 13. and the straw and grains, unhusketl, 
weighed in the air dry state with the following result : 



On the Phvsiolosrical Action of Iodine and Flnorino Compounds on Aflrric. Plants. 475 




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 10. The total quantity of potassium iodid applied to the 
plot No. I. was o.25g., to tlie plot Xo. II. o.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 somewliat below the actual production- 
The straw and grains, unhusked, were weighed in the air dry state with the 
following result : 



< "ontrol. 



\\'eiglit of straw and grains. 
Weight of grains. 




A small increase of yield had therefore taken place by the application 
of 0.2 T g. KI for 20 ~ Meters, while 0.02 ^g had no influence. 



* The solution was liighly diluted, each dose of j otasr'um io^lid being d!ssolvcil in 10 litres of 
water. 



4/6 



S. Siizulii aud K. Aso. 



B. On the influence of potassium iodid on radisJi 
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. I 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 


10 


Large plants. 








Average periphery' Weight. 


5440 g. 


7500 g. 


2770 g. 


of roots = 9.5 c.m. 








VWe!ght of roots. 


2370 „ 


3360 .. 


1440 „ 


/Nuiiilicr. 


24 


20 


15 


Middle sized plants. 








Average periphcryj Weight. 


4020 g. 


4220 g. 


3020 g. 


of roots = 7.5 c ni. 








V Weight of roots. 


1370 „ 


1540., 


lOIO ,, 


/Xuniliur. 


17 


17 


35 


Small plants. 








J'eriphcry of roots-' \\cight. 


i96og. 


1980 g. 


3 nog. 


--4.7c.m. and less. 








MVcight of roots. 


520,, 


510,, 


790 » 


Total weight of plants. 


1 1420 g. 


13700 g. 


8900 g. 


„ ,, „ root. 


42^)0 g. 


S4iog. 


3240 g. 



> Ivich plot was manurcil with 200 g. double supcrphosph ilc, 312. 5 g. ('^H^), ^' \< •>'"' 3'2.5 g- 
wootl ash, the latter being given in a highly diluted state ti.-M days later. 



On tlie riijsiologieal Attiou of loiliup and Fluorine Compoimds on A^ic. 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 = 0.05 l;', 

Corresponding for i ha = 25 g. 

its value = 0.6 yen 

The increase in harvest per 20 sq.m. = 2170 g. root, 
Corresponding per ha = io85000g. ,, 

= 289 Kwamme. 
its value — 2.89 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 graduall}' 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. Oil the influence of sodium fluorid on oats. 
By K. A so. 



In a former article was shown that fluorine in the form of sodium 
fluorid applied in exceedingly high dilution on barle\-, wheat, rice, soy-bean 



1 Since tliis ash contains about 5 per niiilc iodine, 5 Kilo of it would suffice to supply the necessary 
quantity per ha. 

* Bui. College of Agriculture, Tokyo, Vol. 5. Xo. 2. p. iSi. 



I 



4/8 



S. Suzuki aud K. Aso. 



and pea plants, can exert a stimulant action.- 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 iodic! by S. Suzuki; hence the reader, 
is referred in this regard to the introductory remarks of the above 
communication. 

Pot No. I received on five days o.oig. sodium fluoric! in loo.cc. water. 
No. II. o.ooig and No. III. O.oooig, while No. IV served as control. The 
applications of tlie higlily diluted solutions of sodium fluoric! were made on 
Marcli II, April 14, 21 and 28, and May 6. On May 20, it was noticed 
tliat the plants of No. II. developed best, tlicn followed tliosc of No. I. 
There was hardly noticed any difference between the plants of No. Ill, and 
No. IV. The color of the leaves of No. I. was a little paler than that of 
tlie control plants. On May 29, the number of ears was : 

No. 1. 3 

No. II. 4 

No. III. 4 

No. IV. (control). 2 
The plants were cut on July 6. 'I'lu: straw and grains, unhusked, were 
weighed in the air dry state witli the following result : 





I. 


H. 


HI. 


IV. 


Number of stalks. 

Wcij^ht of grains, un 
huskeci, j^., 

\\ cii^ht of straw , t;. 


8 
23.0 
50.1 


9 

24.2 


10 

25-5 
4S.6 


9 

21.4 
45-2 



This result undoubtedK' shows a stimiflant action of fluorine in the 
{proportion of2.i7g. in 10000 kilo soil as in No III, altliough tlie differences 
arc: here not so large as in the case of the i)ea, described in .1 former 
I lulletiu. 



2 Altliou^,'Ii llu,- prcsc-iin- 1)1 lluorlni.' lias to hr a-suimd aliiK st in every soil, it is ol's|)ec'al interest 
that it occurs naturally in wines tVoni ci rtain countries (1 lol/.nian). 

> CI. I'.ul. \'. Xo. 2. 



On tlio Phjsioloj^ioal Action of Iodine and Fluorine Componnds on A??ric. Plants. 479 

D. On the influence of sodimn fluorid on radish. 
By K. A so. 



T\vo plots, each measuring" lo square metre, had received during the 
summer, o.6g and 0-o6g NaF., and again shortly before sowing the seeds of 
radish, they received 0.8g. and o-oSg. sodium fluorid respectively. The 
manure was the same as mentioned in the above field experiment with 
potassium iodid. The radish was sown on October i, 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 stimulating action of considerable magnitude is therefore quite 
evident and it is of special interest that the smaller quantity .o.i4g- 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 : 

NaT applied for 10 square metres = o. I4g. 
,, corresponding to i ha. — 140 g- 

Its cost = 8.4 sen. 

Increase of harvest [)er 10 sq.m. = 4. 1 56 Kg. 
,, corresponding to i ha. = 4156 Kg. 

Its \alue -- 1 10.6 ven. 



I 



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 
Slozijtzoff^ d.x\d o^ Epstein- seem to fully establish the enzym-nature of the 
oxidases. Recently, however, J. H. Kastic and A. S. LocvcnJiart'^ 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-oxiciizable 
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." 

* Z. rhysiol. Chem. 31. Slowtzoty ohs-cwcA that ilie action of l.iccase is proportional to tlic square- 
root of its quantity. He considers the laccase as a peculiar protein substmice which is not changed l>y 
pepsin or jiancreatin. In the pure state, it is killed already at 5o"c, in presence of mineral substance 
however between 65-7o^c. 

2 Arch. Hyg. 36. p. 140. Kpstdn observed tliat 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 .\meric. Clieni. Jourii. WW 1. N>\ 0. Dec. looi. 



4^"^ 2 K. Asc. 

These authors based their view principally on the following special 
observations : 

I. Benzoyl-phthal}-l-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 anchors 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, jDotassium ferricyanid), but also oxid of 
silver and further (|uinone 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 
phenylh}'drazine 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 h)'drocyanic acitl on the action of oxidase was 
obser\ed again by Kastlc and Locvcnhart- after Epstein and also myself 
had made the same observations. 

A furtlu r observation of those authors relates to the inhibiting action 

N 

of sodium thiosulfate. When to 2C.c. of a potato extract o.sC.c. of 

' ^ lOO 

solution of tin- thiosulfate was added, tlie l:)lue color with guaiiicum was 
])re\'ent(.:d. Since other (-:nz)'ms are not injured 1)}' sodium thiosulfate, it was 
inferretl that the oxidase is no cnz)'m proper. Hut here- it might be objectetl 

■ <,>uiiiiinu iJroducL's ;i Muf color also Willi Hit- tctra ])a|)cr of Warslcr. TIk- cnninioii (|iiiiu)nc is 
li<'lial)ly .1 (liUctoii ami imt a |icToxi(l as formerly hc-liovcd (Fitli^,'). 

- They oljscrved tliil '-9 parts prussic aciil in 10 million parU of juice very nearly mark the limit 
ot the poisonous effect of Ih it acid on tin- oxidizint; sul)slances in the potato. 



On tlio ( heuiical Nature of tlie Oxidases. 4^3 

that an oxidizing cnzym must naturally contain diffcrenth' 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 3'ear 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 Ijlue color after it has made its appearance. - 
Such facts might also apply to the observation of Kastlc and Lvcvcnliart 
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 first 
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 slowl}' produced with the juice itself. 

Recently Each and Chodat observed that the juice of LatJiraca 
squainaria 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 \-on cinem ac\'Iirten Ih'droperoxid herrlihren." 



1 A similar oI)servation was iiv.ule liy the w riter in rei^anl to the behavior ot onzyms toward 
sodium tluorid ; while most enzyms are not injuretl by it, the oxidase proper (laccase) is killeil easily. 

2 Compare my article, • On the Role of Oxidase in the prejxaration of Commercial Tea ; l^ul. 
Colle;^e of Agric. Tokyo, \'ol. I\'. No. 4. p. 256. 

3 Schonbcin had observed as early as 1SO4 (Jonrii. liir prakt. Chcm. lid. SS.3.460) that certain 
aqueous extracts of plants give a blue reaction with acidulated iotlid of potassium starch, which iwaction 
he supposed to be due to nitrous acid. Many ]ilant juices however yielded that reaction only after 
standing for a series of days. In the latter case, nitrite might have been produced from the nitr.atos. 
frciiuently 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 volume of a 2% starch paste to which 

I ^^ potassium iodid and 0.5 % acetic acid was added. These mixtures 
)'ielded even after twent)' four hours standing in darkness, no trace of any 
blue reaction, while the guaiacum blue reaction even in absence of h}'drogen 
pero.xid was still obtained with great intensity.' 

J>ac/i and Chodat recommentl to add some mangano-sulfate in those cases 
in which the iodine reaction with ])lant juices fails. Ikit in the above 
mentioned cases with the juices of jjotato and r.idish, this sulfate ditl not 



» Til one case I \\m\ apjiliol iiitciitiDimlly a polassiuni ioiliil solution not freshly piepareil, but one 
which had been exposed in presence of air for a few days to sunlij,dit. In this case, a blue reaction was 
j^'radually ol)-;erved, evidently due to slight traces of free I'odine formed in this solution. 



I 



On the Clieiiiical Nature of the Oxidase-;. 4'^ 5 

change the result. However after sucli 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 i% 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 h\'drogen peroxid of about 2% and of a faint acid 

10 ^ to r 

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 /lo reaction ivJiatcvcr i^uis taking place. If Kastlc and Locvcnliarf s 
view v/as correct, then the supposed organic peroxid would be formed 
almost instantaneously when hydrogen peroxid comes in contact \\\\.\\ the 
proper organic material in tlie juice. This supposed organic peroxid would 
consequently be also present in the alcoholic precipitate containing all the 
oxidizing enzyms, hencc 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 : no reaction in absence, but an intense reaction in 
presence of hydrogen peroxid. What is true for this kind of peroxidase 
Q^-guaiacolase) is very probably also true for the common peroxidase 
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, 

i Since Boiiiquih^t ol)scrvc(l in the funy:us Kicssu/j, an oxidizing enzym which produces a retl 
color with guaiacol even in absence of hydrogen peroxid, I propose to distinguish this peculiar enzym as 
a-guaiacolasc, from the above-mentioned enzym which T call /3-guaiacolase. Ai)Out this reaction 
compare also my article, ' On ( ixidizing Enzyms in the ^'egetal>le Body ' l?ul. College of Agric. Tokyo, 
Vol. V. No. 2. p. 207 — 235. 

- On heating the solution of the enzym precipitate above mentioned for 5 minutes 10 75', the 
oxidase ami the comm-):i peroxidase are killed, while the guaiacol-hydrogenpcroxid reaction was still 
obtained a'lhouch 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 pcroxids arc then tlic first products of an 
oxidation caused by the oxidizing cnzyni and this opinion seems to be also 
that. of Bach and Chodat, and differs essentially from the hypothesis of Kastle 
and Loevoihart, according to ivhich oxidases themselves are the pcroxids. 

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 diethylperoxid nor dibenzoylperoxids will liberate 
iodine, but benzoylliydroperoxid 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 
Jiydroperoxids 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 im[){)rtant to tlecide 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 prtu'e of some interest in 
regard to this ciuestion. 



I RfCi-'iitly, A'. //. I'iige (Aiiier. I';it. 717016 of 30. 1 Vc. 1902) (locrihcd iiCLtyihyihuptToxid 

ellj,— C_Q_f ,_l I „|,i^.], ],.,^; ^ strong odor after liy[otliliiioii-; acid ;iiul l)ii>- a lowcrt'ul luRk-nVida 
action. 

* (omiarc in regard to the data liere mentioned, tiie articles of i^n.ycr and I'illiger in IJericlitel 
iler Deiitsclien C'liemisclien Cesellscliat't, 1899 and 1900, especially in the latter volume, page 157S. 



Oil tlio Chemical Nature of the Oxidases. 487 



Experiments i^'it/i Buds. 

Sections of potato-buds yielded directly the iodine reaction in presence 
of some acetic acid.' Also the blue i^uaiac reaction was directly produced. 
The cold prepared extract behaved alike. In a second case, with buds 
from other potatoes, however, the iodine reaction failed, althouijh oxidase, 
peroxidase and ■i'-guaiacolase- were present. Of considerable interest were 
the observations made with the tubers and buds of Sagittaria sagittcefolia. 

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 Sagittaria were extracted with 
looc.c. water ; a portion was tested directly and another after boiling for 
^/^ minute. In the former case, oxidase, peroxidase and ,5-guaiacolase 
were easily recognized b\' 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. Xot only the unboiled, but also the boiled 
juiee yielded this reaction with great intensity after addition of some acetie 
acid.^ Even boiling for 2 minutes did not alter this result.^ 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 buds. Hence four bulbs 
were crushed after removing the skins and macerated with lOOc.c. water. 
The filtrate was mixed with alcohol, whereby a consitlerable precipitate 

* The tubers did not t^ivc this reaction, as was mentioned aliove. 

2 See above p. 485. 

3 A l)!inil control test witli acetic acid and jiotas^iuni iodiil-stracli [laste showed no reaction 
whatever. 

•» Bach and ChoJat mention that heating to So^C prevents the iixline reaction. Tins is, however, 
probably only the case when the acidity of the juice is more marked than in the case of the Sagittaria 
buds. It can then be very easily explained that nirons acid set f.ee reacts upon amido-compounds 
and is destroyed with development of nitrogjen. 



488 K. Asn. 

was obtained. This was filtered oft', 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 solu?jle albumin, it was not surprising to find that the juice 
of the bulb was capable to prevent the iodine reaction with the juice of the 
bud, and further that the boiled ]\\\ce 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 }^ield 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 Gricss for nitrites yielded at once a very decisive result. Ilence the 
liberation of iodine is due not to any enzym nor to any peroxid, but to 
nitrites. 

It is ver)' strange that the occurence of nitrite in plants thus far was 
overlooked. It is true that ScJionbciu more than thirty years ago had 
supposed the existence of nitrite in plant juices and further that BcrtJiclot 
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 
poison(^us for plants with an acid plant juice.' But in this regard we must 
not overlook that the (juantity 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 r)f these shoots is exceedingly weak. 

1 O. I/)c-\v, Naturliches System dcr Giftwirkuni^cn, p. 61 and p. 109. 



On the Cheinif al Ji'atnie of the Oxidases. 4^9 

Since the question is of some interest whether this nitrite is formed by 
reduction of nitrate or by oxidation of ammonium salts, I hav^e 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 tliis 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. 



Sununary. 

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 he prevented by the presence of certain I)c:'.zene 
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 phsenogams but they are directly poiso- 
nous.^ 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— yS hydroxylamine- 
disulfonic acid which preparation was kindly furnished me by Prof T. Ilaga.- 

This compound has the formula. 



N^ SO,. Na 

^^\0. SO^. Xa. 
I prepared the following solutions : — 

a) I per mille Calcium acetate. 

I ., ,. Magnesium sulfate (anhyd.) 

I ,, ,, Potassium chlorid. 

I ,. ,, Ferrous sulfate (anh)-<.l.) 

b) 2 per mille Dipotassiumj^hosphate. 

I ,. ,, Sodium a ■ /5. disulfln'droxylamate. 

and for the control plants : 

c) same as a). 

1 Locw, Kill iiatiirliclies System ilcr (."littwirkungeii. 1S03, p. 41. 

2 This interesting salt was prcjiarcd by Prof. Haga from oximiilo sulphonatc o( >i-Kiiuin hy a com- 
plicatctl process which will be pubb'shcd later liy that autlior. 



49- ^' Suzuki. 

d) 2 per millc Dipotassium phosphate. 

0.28 ,, Ammonium sulfate.! 

The apphcation 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 cm. 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. lOth, 
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. lOth, since tlicre 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: — 



Solutitjn of 
A 



Numl)ci" 
stalks, 


of 


Number 
living 


of leaves, 
dead 


Length of the 
longest leaves. 


Weight of the 
fresh plants. 


d b series. 












1 .. 




-, 


(') . . 

6... 


13 cm. 

II 


1 
I 




1 .. 




■ ■ ' :> 


2.68 g. 


I . 




1 


6... 

5 ■■• 


12 ,, 

II 


1 
( 




I .. 




-7 


2.00 ,. 



* This ainmount of anm.oniuni sulf.ite cori-esponds to lli.il of the solium salt of o /3. hydioxyl- 
aniine disulfonic acid, in the solution (h), in regard to the amount of lu'trogcn. 



Can siilfo-dcrivati ves of hydroxylainiiie servo as a source of nitrogen for plants. 493 

Control plants. 

,. ( I 5 12 5 17.5 cm.) 

L \ r 7-02 v;. 



4 7 4 I/O 



-. f 



D 8.42 ,. 

^ 4 9 -4 1(^0 .. 



Experiment ivitJi f/in^qi. 

A culture solution was prepared, consistint; of 
200 c.c. Water. 

2 g. Canesugar. 
0.4 ,, Hydroxylamine disulfonate of sodium. 
0.2 „ KH3 PO4. 
0.0 1 „ Mg SO 4. 
Two flasks, a and b, each containing 100 c.c. of this solution were after 
sterilisation infected with : 

a) Pcnicilliinn glaucinn. 

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. j3. hydroxylamine disulphonic acid is no direct poison for the 
barley but it is incapable to furnish nitrogen and hence the plants un,dergo 
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-yS-hydroxylamine tlisulfonic acid. 

1 It is of some interest to compare with this result tlic loisonous character of ainiilo-sulfoiiic acid, 
these Ikilletins, vol. II. pai^e 4S7 {1S97). 



On the Influence of a Certain Ratio Between Lime and IVlagnesia 
on the Growth of the Mulberry-tree. 



I5V 



K. Aso. 



Since sericulture is one of the most important ay;ricultural 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 reviev.' 
of some of these may be not out of place. 

The composition of the ashes of health)' mulberr\'-lea\-es was found in 
average, as follows :' 

l\ O. 12.02Q^ 

1^2 O 3^-47% 

Na,0 3.140^ 

CaO jj.is% 

MgO 12.48% 

SO, 4.64^ 

CI 0.06% 

SiO, 1.45% 

1^^.03 1.59^ 

On analysis of the bark of the health}' mulberr)'-roots (var. Xezumigae- 

shi) collected on Dec. 4., 1 have obtained the following result: 

In 100 parts of the crutle ash, 

?„ O. 18.4S 

Ko O 9.39 

CaO JS-jo 

MgO 7.34 

' N;vj;;u>ka : C'l'cniical TaMcs tor Daily I'm', p. S4. 



496 



K. Aso. 



s o. 

Si O. 



Fcg O3 



1.38 
2.40 

8-73 



U. Suzuki ^ has determined the hme 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 
greater in healthy than in diseased leaves. 

?iIaeno2 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 develoi)ment. My analyses, 
indeed, have shown that the amount of magnesia iM-ed(-)minated over that 
of lime, wliich is a ver>' unfavorable condition. 

In 100 parts of dry soil, 



I.oC.M.lTV. 


Ca 


.MgO 


•Jclakaramura, (Aicliiken) 


0.232 
0.1 15 
0.150 


0-332 


Jutaii Sciicultiiral Scliool, (Kyuldfii) 

.\iij^;iinui';i (Kyuttiiu) 


0.250 
0.3S8 







J lliil. Collfj^c cif Aj;iicullurc', \>A. 1\ . .No. 3. 
' Il)iil. Vol. TI. \o. 7. p. 495. 



On tlie Ijifliionoo of a cortaiu Ratio ])(4wcon Liino and Magniesia. 



497 



The following experiments will prove, indeed, that a normal and good 
development of this plant depends to a great extent on the ratio of lime 
and magnesia offered to the roots. 

Experiment with U^ater Culture. 

Three young mulberry plants (var. Takasuke) with stems about 15 cm. 
high were placed June 9, in glass vessels of 3 litres capacity containing the 
following solutions : — • 



Ca (X03)3.. 
Mg (NO3}, 

(NHJ, SO, 
FeSO 



11. 



III. 



o-5% 
0.1% 
trace 




0.5% 
0.1% 

trace 



On July II, there was not yet any other difference observed except in 
the number of rootlets that had developed. There were very numerous root- 
lets in I and II, while none in III. 

On July 25, it could be clearl}' noticed that in solution III, the leaves 
developed were very small and of pale green color. On August 8, these 
small leaves had withered \\hile those developed in the other two solutions 
appeared healthy and of dark green color, but it was further noticeable that 
the leaves in I were darker green and smaller than those in solution II. 
Further, while no rootlet were developed in solution III. numerous rootlets 
had appeared in I and II. This experiment shows that an excess of mag- 
nesia over lime is \^er\' injurious ^(^x the mulberr\' tree. 



Rxperi)uent ivitJi Soil Culture. 

l^ach pot contained about 3.8 K. ilry soil of an unmanureil field from 
Nishigahara, Tokyo. The content, in the fine earth, of lime antl magnesia 
soluble in hot concentrated In'drochloric aciil was as follows : — 



49<'^ 



K. Aso. 



In TOO parts of clr\' soil, 

Ca O 1.5 

Mc^ O 1.8 

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 witli it to reach the 
following" ratios : — 



Pots 
a 
b 
c 
d 
c 

f 



Ca O 
I 
I 
I 
2 

3 

4 



Mil O 



I (original! 



The surface of the pots measured 0.0495^ m. As general manure 
served : 

7.5 g. N in the form of ammonium sulf.ite and 

5.6 g. Po O.-, in the form of potassium i^hospliate for each pot. 

These salts were applied in solution. Young mulberr}' plants (var. Shi- 
hozaki) of equal size, weighing 976.7 g. — 1014.3 g. and of stem-length of 
about 30 cm. were planted on April 21. 

On June 6. and Sept. 19, the following obserx'ation was made: — 



1 )ale. 


1. 


11. 


III. 


n. 


\' 


VJ. 


June 6. 


Leaves 
very small. 


Leaves 
small. 


Coiilrol. 


Developed 

Let. 






Sept. 19. 


One jilant 
died. Wilh 
another the 
leaves are 
very small. 


Leaves 

fleveloped to 

sdnic extent. 




I )eveloped 

very well 

nearly, same 

as T\'. 


Ix^ss well 

developed 

than \'. 



Oil tlie Jiifliiciicc <>r ji tcrtaiii Ratio bchvocii Lime and Maj^iiesia- 



499 



On Oct. I. A photograph was taken (plate XXVIl) which exhibits the 
great difference of devcIo]M-ncnt at once. (3n Oct. 2 the following obser- 
vations were made : — 



No. of 


CaO 


Number 

of 

leaves. 


Fresh weight of 
total leaver. 


Average 
A\eight of 
one leaf. 


Number 

of 
l)ranches. 


Remarks. 


jiol. 


MgO 




I. 


0-33 


8 


1-5 


0.19 


3 


15i-anches were 
very small. 


11. 


05 


16 


6.8 


0.41 


4 




in. 


I 


21 


9.8 


0.45 


7 




i\'. 


2 


30 


31-9 


1.05 


7 


One branch was 
longest of all. 


V. 


3 


3^: 


3f'-4 


0.9S 


8 


Average development 
of branches was here 
better than hi 1\'. 


M. 


4 


20 


135 


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 on.e branch more, it will be safest to conclude 
that the best ratio Ca O : Mg O for the mulberry tree lies between 2 and 3. 
If follows further that an excess of magnesia over lime dejiresses the growth 
considerably ; the leaves become smaller, but true symptons of dwarf- 
disease are not observed. 



On the Influence of Different Ratios between Lime and Magnesia 
upon the Development of Phaseolus. 



BY 



G. Eaikuhara. 



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 Lociu, May, A so and Funtta. O. Lociv has named the ratio 

Ca O 
of-^Tr — ?=r 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 Ci '3 +>!;; (-'^a 
for the air dry sand : 


A. 
0.05% 


r.. 

o.i9o 






I 


_3_ 

I 


3 


3_ 

I 


l.imcfactor : 

CaO 

MgU 


11 


2 
I 


2 
IT 


I 


III 


I 

I 


I 
I 


I 
I 




IV 


0.5 
I 


0.5 
I 


0.5 

I 




\- 


£i33^ 
1 


0.33 

I 


0-33 
I 



50- (t. Daikuhaia. 

As t^cneral manure for each pot served : — 

K„H PO, o.i^ 

K H„PO, o.i% 

KNO3 0.2^ 

(NH,),SO, 0.1% 

P^'c SO^ 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 tlie ratio -^r^ — 7-.- was = 2. 

Mg O 

The following table gives the measurements taken at this time : — 





CaO 
Mg 


A. 


B. 


C. 




3 


13-5 


1 1.5 


135 


Length of «tcm. 


2 


18.5 


17.0 


»5-5 


cm. 


I 


14.0 


14.0 


1 1.5 




0.50 


13-0 


15,0 


lO.O 




0.33 


13.0 









3 


S.o 


6.0 


5-0 


Li;iii;tli of tlic largest 


2 


9.0 


7-3 


5-5 


leaf, tm. 


I 


7-5 


6.0 


55 




0.50 


6.0 


6.5 


50 




0.33 


(>-5 








3 


3-9 


4.0 


3-0 


llreadth <if tlie largest 


2 


40 


3S 


32 


leaf, cm. 


I 


4.0 


30 


30 




0.50 


2.S 


35 


2-5 




"•3."> 


3-5 








On the Inducco of l>im.„.«t Ratios l,,t,vee„ Uu,. and Magnesia. 503 

Tltis tabic shows clcarlj- not only an /„/„..„,r ,/ r/u- r„ii„ .fSlSL 
„pon the ,.ci,J, „f tlu- riant l„„ also on thr sir.- of tlu- Ica-oos. ' xtL 
ratto ,s korc . .• ,, at least before the fruiting stage of this plant. The 
plants No. V of B and C had died at the time the measu,e„,ents were n,ade 
very probably from the excess of magnesium carbonate. 

Unfortunately the experiment had to be terminated soon afteruards 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. 



Man}' investigations have been carried out to determine how much 
phosphoric acid in a soil is avaih^ble 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 i^^ 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 o.oi percent of phosphoric acid soluble in 
a I percent solution 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 otlier acids 
in this regard and also to compare different soils. 

I. Application of organic acids in i per cent solution. 

The samples of soil were taken from the experimental paddy field 
(sandy loam) of the Kinai Branch of the Imperial Agricultural ICxperiment 
Station at Kashiwara, Osaka, which were manured every crop with difierent 
quantities of phosphoric acid during three years as follows : — 
No. T. No. \\0^ 
-I No. II. 37.5 Kg PjO. as Superphosphate per ha. 



I'addy 

Soil. 



No. III. 93.75 Kg P,0- „ „ „ „ 

( No. I. No. TjCV,. 

I No. II. 37.5 Kg V^O. as Superphosphate per h.i 

^ No. in. 1 12.5 KgPjO. „ „ .. .. 



1 Journ. of the Clieni. Soc., London, 6<, 11^ (1S94). 



5o6 (t. JMiikiilinra. 

Each plot had received moreover 112.5 Kilo N as NH_jCl and 93.75 
Kilo KgO as K^ CO, per ha. 

The extractions were carried out according- to B. Dyer's method. The 
followini^- table shows the result of analysis in the percentage of dry fine 
earth : — 

r,( )^ Solul)le ill 



1% acetic a. i% tartaric a. 1% citric a. 1% oxalic a. 

/ Xo. 1. No. l'.^Oj. 0.00832 0.04190 0.08381 0.12955 

^■"' J No. 11. 375KRr,()- 0.04798 0.09596 0.16939 

Soil. 

[ No. IIJ. 93.45 Ks ^2*^^-, 0.011S4 0.0607S 0.09916 o.iSooi 

/ No. ]• No. l',< '5 0.00096 0.0092S C.01S23 0.04606 

^''^1 No. II. 37.5 Kg 1*2' >-, 0.0012S 0.0140S 0,02495 0.05342 
Soil. I 

No. 111. ii2.5KsT,0- 0.00192 0.01823 0.04526 0.09660 



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 : — 



On the Boliiivior oftlic riiosidioiic iicid in the Soils toivards diffcroiit Acids. 50/ 





^. 


^O 


Vo 


N= 




vo 


\o 






'u 


c\ 


C\ 


=^\ 


c" 


c\ 


c\ 


C\ 




rt 




C/0 


M 


■+ 


lo 


<s 


-r 






o 


\o 


O 




1^ 


•+ 


W-5 




Z::^ 


O 


r^ 


u", 


-t- 


O 


■^ 


o 




ri 


o 


^ 


o 


q 




^ 


o 






d 


d 


d 


C 


d 


G 


d 








































-e 




^ 


c -, 




■■ o 


^ 


C X 


^ 


r^ 


LO 


CO 


00 


u-i 


O 


\j^ 


I-'". 




t-l 


•q- 


Tl- 


•1- 


CO 


r-- 


t^ 






o 


fl 


M 


00 


-^ 


8 


•^ 


'r 


O 


o 


o 


o 


o 


O 


•S 


:- 


d 


d 


d 


d 


d 


d 


d 


•j 




































^ 


















Zj 




































n 


















J. 


— " 
















_o 


'u 


^ 


^? 


^=s 




^ 


^ 


^ 


^ 




c/o 


l/-> 


O 


r<^ 


M 


c> 


—J 


?* 


'^ 


o 


o 


t^ 


ro 


ro 


^ 


^ 


" 


'b 


8 


? 


o 


O 


CO 

O 


O 


>. 




T^ 


d 


d 


d 


d 


d 


d 


> 




r^ 








































vo 


o 




\o 


NO 






o 










c\ 


c\ 






rt 


o 


l-O 


,^ 


C-» 


O 


CO 


^ 






rj 


u-, 


'~ 


u 


*« 


"1 


o 












r; 


8 

d 


8 

d 






u 


- 


d 


r^' 


■= 


- 




<;■ 
















^ _a \\ 


^ 




^ 


No 




C\ 




3' ^ r- >— 


r^ 


O 


a; 


O 


00 


vO 


r'^. 


.-* ^ •— ""^ 






o 


•-' 


*^ 


►-• 


r') 


~ X o 


d 


C 


d 


d 


6 


° . 


d 


















































































3 


>^ 


>> 


o 


tA 


rt 


^• 


>!, 


u 


<L> 






o 








rt 


>^ 






>-^ 








2 


C3 




^ 


"0 




c 


^ 


'-^ 






rt 

in, 




rt 






































.y c 

tr. -^ 


_5 


1 


, 


^ 


„ 


^ 


5 




^ 


5 










3 




































































=■ 














X 


rt 






























^jLw 5 


cT: 


— -■ 


-^.- 






_^' 
















u 




^ rt 


r3 


"^ ^■ 


^ o 




^ 




;^ 












J 








O 


15 "^ 


1 =^ 


y 


rt 


^ 


»*^ 


C '^ 


O 


Js Cl, 


-^ ^ 


_^ 


^ 


D 




^- ,=? 





" D 


'"' ;il 


^^ 


o 


^ 


5 


















p^ 


^^ 


rj 


rt 


~ 


>^ 


_a: 


^ 






]o 


i5 


|_^ 


'r^ 


:>: 


X 


^ 



5o8 



(i. Daikiihara. 



II. Extraction of Soils zvitJi Organic Acids of Different Strength. 



The soil serving for these experiments contained 1.635^0' of hygroscopic 
water and 0.1 727^^^ of Po O- sokible in boiHng hydrochloric acid (sp. gr. 
1.25). The method of extraction was also here that of Dyer. 

The following table shows the results : — 



Df the acids. 


Acetic acid. 


Tartarc acid. 


Citric acid. 


Oxalic acid. 


0.25% 
0.50% 
i.co% 

-OO/o 

5-oo% 






0.0855% 
0.0948% 
0.1029% 
0.1173% 


0.1326% 
0.1609% 
0.1798% 
0.1954% 
0.1964% 


0.0355% 
o.o499''o 
0.0586% 


00403% 

0.0495%^ 
0.0704% 
0,0918% 



In this case the extractive power of oxalic acid for Po O- in the soil was 
strongest, next in order came citric and tartaric acids, and finally acetic 
acid. 



Can Boric Acid in High Dilution Exert a Stimulant 
Action on Plants ? 



I'.V 



M. Nakamura. 



It has been shown by various authors that the soil of certain districts 
contains small cjuantitics of borates, hence also the plants grown on such 
soils contained some boric acid. E. Hotter'^ proved the presence of boric 
acid in many plants by extracting their ashes with water and transforming 
the boric acid into its mcthylic ether which was distilled off. It is especially 
the fruits in which the boric acid accumulates ; in lOOOO parts of the dr\' 
matter of fruits the amount of boric acid w'as found to var}' from 2.2 to 12.S 
parts. 

(^r?///j'<'7//- made similar observations. Of some interest is also the obserwi- 
tion of Craiiipton'^ that boric acid occurs in grapes grown in California. 
A. Hcrrjfcld and IC. v. Lippmann have made further observations on the 
occurrence of not insignificant traces of boric acid in lemons and other fruits. 
/''. ScJiaft'cr^ 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 ver)- much injured and the plants ilietl 
after 20 days. l-Aen an amount of 10 milligram boric acitl jier liter can 
exert some noxious action ; some difference in resistance power was, 
however, noticeable with various plants.^ The source of boric acid in the 
soils is in'obabK- turmalin which mineral contains about \0% of boric acid 

1 JahrcsljcriclU f. At^ricultur C'lioniic. 1S90. p. 203, als;) /.L-itschrift lur Xalirunijsmittc!, etc. lb'95. 
'■^ Jaliivshcricht 1". Agricultiir Clicinic tS()0. 
■^ Jahrcslicricht f. AgricuUiu" C'hcniic iSSo. 
* Schwciz. Woclicnschr. Chcm. I'liarni. [iqo2,] ./i', p. 47S. 

■\ In rci^aul to algae (Spirogyra, Naiiclicria) I.oew mentions in I'lora, 1S02. p. 374 ll'.ai lliey arc not 
injured \\itliin several weeks hy adding 0.2 per niillc horic acid to the culture water. 



5 lO Can Boric Acid in High Diintldii Exert a Stimnliutt Action on Plants .' 

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 lo milligr. and 50 milligrams respectively of borax 
per Kilo. These pots were manured with ig. XaN03, ig^"- 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 
15 cm 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 2 5"C. Measurements of the shoots to 
the tip of the longest leaves were made on Nov. 9, Dec. i and Dec. 12 with 
the following results: 

Measurements Cm. 





Nov. 9 


Dec. I 


Dec. 12 




, I 


31 
27 
26 

2S 


39.8 
35.5 
34 
38,5 


45 




1 


4- 


50 mg 


^ 


41 


I lorax 


4 






46 




Average 


28 


36,8 


43.5 




f I 

2 


30 
24 

33 
2S 


41 

37 
43 
37 


51 

4i 






51 


Control 


4 




40 




Average 


29 


39.5 


46,75 



Tile percentage of increase was therefore from Xo\-. 9 to Dec. 12 the 
following. 

I ' ) .•) ' ^0 I 

50 mg Horax ...J "■' 35'7/o [. average = 35,5% 

4) 39.0% 



M. XakaiiMira. 



511 



Control. 



15.3%' j 



average = 38, 5% 



1) 41,2% 

2) 46,6% 

3) 0; 

4) 30. 

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 ]\Iarch 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 \vt. 


Number of 


Number ot 


Average length 






grains 


branches 


of branches 


Control 


450 g 


132 


8 


63,5 era 


10 mg. Borax 


28,2 „ 


30 


4 


58,0 „ 


50 mg. Borax 


17,3 >. 


24 


4 


46,0 „ 



In the following experiments, commenced February i, with pea and 
spinach the amount of borax was reduced to 5mg. and img. per kilo soil 
respectively. 10 seeds 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 


qnii:. Borax per kilo soil 


62 cm 
86,25 „ 
69,5 ,. 





ini'T. Borax per kilo soil 


6 


Control 


3 







512 Can Boric Acid in High Dilution Exert a Stimulant Action on Plants ? 







Spinach 






Average vvt. of plants 


Average length of leaves 


5mg. Borax per 


kilo soil 


1 0,3 5 g. 

7.2 g. 


38,2 cm 
34,0 „ 









It will be observed that one milligram of borax per kilo soil exerted 
some stimulant action with the pea plants and 5mg. 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. F. Hofvianii' inferred from his experiments with dogs and 
rabbits that boric acid is " ein starkes Zellgift." Rosf- observed that the 
body weight decreases continuously by the use of borax and vomition and 
diarrhea may result. E. Kistej-'' and also G. Mcrkcl^ observed that 1 — 2 
grams of boric acid can produce injuries of the stomach and diarrhea. Such 
an opinion was also expressed by H. Maycj-J' On the other hand Liebreich 
and Gcrlach deny the injurious charactar of borax and boric acid in small 
doses. However, when wc 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). Metl. Wochen?clir. 1902, No. 46. 

2 Ibid. 1903, I'ebruary. 

=» Zeitschr. Ilyg. 1901. 

•« M. Med. Wochenschr. 1903, No. 50, p. ico. 

i^ Hyp. Rundschau 12, 1230. 
It must also 1)0 ment'oncd here that Doane and Price reported that c.dvcs ted with milk containing 
borax lost their hairs Marryland Agric. Exper, Station Report No. 86. 



On the Action of Vanadin Compounds on Plants. 



KV 



S. Suzuki. 



Although vanadin compounds occur \'cry rarel}- in nature, vanadin was 
nevertheless discovered in the ash of the sugar beet by Ed O. v. Lippinann. ^ 
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 the}' 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 cm. high) were placed in solutions of vanadin sulphate of i^q ; 
0.1% and 0.0 1 per cent. After 5 da)'s the shoots in the solution of i^q were 
dead, while in that of o. i^'q the leaves wilted. Rut the shoots in the o.oi^-q 
solution were still heabthy even after 12 days. 

Water cultures in Knop's solution-' were also started to which i.o and 
O.I 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 thircl pot served as 
control. Each pot contained 10 kilo soiH 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. 

' llcrl. lici". \o\. 21, p. 3492. 

- I applicl the bluish grccii .sulphate ot" co;nmLTCe, tlie so c.ille.l hypovanadic sulphate, 
^ •-• ' '2 (^^^4)2- "I's salt has a strong acid reaction. 

3 ( Illy the .iniount of magnesium sulphate was a l.ttle increased. 

* Each pot was niaiiurcd with ammonium sulphate 2.3 g., sodium nitrate 2 g., potassium sulj hate 
2 g., sodium phosphate 2.5 g. an 1 sodium chlorid i g. 



5 14 ^' J^iiziiki' 

Water culture. Barley shoots (16-17 cm. high) Averc placed (Dec. 4) 
in 6 flasks.' 

a and a, received o. i per mille vanach'n sulphate, 
b and bj ,. O.OI ,. .. .> ., 

c and c^ served as control. 
The observations made on Jan. 20 were as follows : — 

lAnirh of Xuniljcr of stalks Number of leaves Length 
thin 



til 


le longest 
leaves. 


thi'c 


a 


16.0 cm. 


I 


='i 


15-5 >' 


I 


average 


15.S „ 


I 


b 


21.0 c m. 


5 


I'x 


21.0 „ 


5 


average 


21.0 ,, 


5 


C 


18.0 cm. 


6 


<^i 


23.0 „ 


4 


average 


20.5 „ 


5 



, 




of the 


living 


dead 


roots. 


4 


9 


2.0 cm. 


5 


S 


1-5 » 



Remarks 



Xo root hairs visible. 

Development stopped ; 

fresh leaves appear 

but the old leaves 

1 .8 .. ] die off. 



17 3 20.0 ,, \ Normal. 

20 3 



18 




Normal. 



A very weak stimulant action on the roots seemed to ha\'e taken place 
in the case b and b^. l^ut in the cases a and a^ the poisonous action was 
so decisive that tlie plants Were no loni^^er obser\-ed. The final obserwitions 
were made on Feb. 27 with the followinL,^ results : — 





Length of 

the longest 

leaves. 


Numl)er 

of thick 

lliin stalks. 


Number 

of living 

• lead leaves. 


Lengtii 
of the 
roots. 


Weight 

fiesh root.- 

ixjrtion i 


in a 
; upper 
^t:^te. 


b 


32.5 cm. 


21 


I 


7f' 


10 


25 


39-7 K- 


27-4 '^,^ 


w 


30.0 „ 


iS 


2 


70 


S 


-5 


37-7 •, 


^S.3 „ 


average 


3'o •. 


20 


'■5 


73 


9 


25 


3>"^-7 ,. 


-7-9 ., 


c 


34.0 cm. 


-» 1 


I 


7^> 


'3 


30 


37 7 >'. 


-5 9 S- 


«^i 


33.-^ •' 


IS 


' 


66 


10 


-3-5 


4::-7 ., 


2S.2 „ 


average- 


33-5 .. 


20 


I 


7' 


12 


27 


40.2 „ 


27.1 -. 



This experiment proxes tiiat in a normal water culture barley is \"er)' 
much injin-i^d 1)\- the addition ofo. I per milk- xanadin sulphate, and further 

The siilutions were renewed on I )er. 20, (an. S. 20, 31 and i'Vli. 21. 



On the Action of Vanadiii CompoiiiKls on riants. 



515 



that when appHcd in the further dihition of o.oi per inille 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 nuniVier of stalks. 




Jan. 23. 


March 27. 


April 10. 


March 27. 


Aj.ril 10. 


/o.i 5?. vanadiii \ 


c ni. 


cm. 


cm. 






I'ot I. sulphate per 1 
V 10 kilo soil. ' 


8.6 


26.4 


.S7.7 


3 


3 


Pot IT. (ig. „ „ ) 


S.9 


30.3 


59-1 


3 


3 


Pot III. (Control) 


9.1 


37-2 


62.S 


4 


4 


Pot IV. ( „ ) 


9-3 


33-1 


63-5 


4 


4 



This experiment plainly shows th.at vanadin sulphate e\'en in a \ery 
small quantities has no stimulatiuL;" action on barle}'. 



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 higli dihition acts poisonously on plants in water culture.^ 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 WaL;ner's pots each containiuL;' lo kilo soil served for the experiment. 
Each pot received as manure : — 

Ammonium sulphate 2.3 g. 

Sodium nitrate 2.0 ,, 

Potassium sulphate i .9 ,, 

Sodium phosphate (cryst.) 2.5 ,, 

Two pots served as control while one pot received o. i i;". potassium- 
ferrocyanid and another i t;-. J<"ifteen seeds of barley were sown in each pot 
on Dec. 13. and the youn^" shoots reduced to 7 of ec^ual size on Jan. 20. 
After a fc:w weeks a decided difterence was noticed in fa\\>r of the pot 
that received 1 t;-. potassium ferrocyanitl. Measurements were made on 
March 7 and 27 with the following results : — 



1 Tlicsc l^ul. Vol. V. No. 



=;i8 



S. Suzuki. 





Average length of the 
longest leaves. 


Average number of stalks. 




-March 7. 


March 27. March 7. 


March 27. 


Tot I. (o.i g. K^ Fe CVp). 
Pot II. (I - K^ FeCyg). 
I'ot III. (Control). 
Pot R'. ( ,. ). 


14.4 c.ni. 
26.8 „ 
12.8 „ 

15/' .. 


31.2 cm. 5 
44.6 ., 4 
37-2 „ 1 4 
3;.. I ■. 1 


5 
5 
4 
4 



The question ariscd whether the favorable effect in pot II was clue to 
the potassuim 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 ]\Iarch 17 with water and the filtrate tested 
with ferric chlorid, but only an exceeding!}' feeble reaction was obtained. 
In a second test diluted hydrochloric acid served for the extraction, but with 
no better result. A control test with loo g. immanured soil moistened with 
a dilute solution of 10 m.g. ])otassium ferroc)-anid showed further that this 
small ([uantity is entirely absorbeil. 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. 



' I'rovious experiments liad show n 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^ aetion of potassium iotlid on agri- 
cultural crops 1 made it desirable to know wiiether 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 /). Dycr"^ on the field of RotJiauistcd, e. g., have shown that 
chlorids are to a certain degree retained by cla}' soils. He writes: "Now 
the average cjuantity of chlorine which fcdls annuall)' in the rainfall at 
Rothamsted, as calculated on observations for 22 harvest }'ears, 1 877-1878 
to 1898-99, was 14.75 pounds." " Yet we see that the soil of plat 5 in the 
Broadbalk wheat-field retains, on the average, within each depth of 9 inches 
down as f^ir as 90 inches, a quantity of chh^rine ecpiiwdent to tliat which 
falls ui)on its surface each year in the form of r.iin.'' In other words, down 
to a depth of 90 inches the soil, though continually subjected to the wash- 
ing influence of the rain, contains a ([uantit}' of chlorids e([ui\alent to that 
falls upon it during ten }-ears, neglecting the very few pounds annuall\- sup- 
plied to it as impurities in the manures." " It would seem that the clay 
enters into some sort of combinatii")n with the chlorids from which the\- are 
only dislodged by a very free application o{ water." " The ilifficulty of 

* Tlicsc Ikilk'lins \"ol. V. No. z aiul p. 474 in this number. 

" ( 'nice of I'xpcn'iuonl Slatinns, Uul. No. 106, I'. S. IVpait, of Aj^ric. p. S2 and S3. 

=' 'I'licsc ijuantitios arc in corlain countries Comparatively large. Thus in Barbados were found by 
Albuquerque per million parts of rain water from 6 to 3S.5 parts of chlorine, while the nitrogen as am- 
monia varied between 0.015 ^"^ o.i\2. (Report of tlie Aerie, work in Barbados, Government Exp. 
Station, 1902.) 



520 S. Suzuki. 

removing chlorids from the soil by percolation except when a relatively- 
very lart^e quantity of w.iter was used, was demonstrated in some experi- 
ments described in the pai)er 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, i per millc solutions of 
both these salts serving for the filtration through the soil. i\s reagent for 
iodine served starch paste to which freshly neutralized hydrogen peroxid 
and a trace of ferrous suli)hate was added. ^W this reaction of ScJionbcin 
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 cm. high and 5.8 cm. wide. 200 cc. of 
each solution were poured gradually on the surficc of the soil contained in 
a cylindrical vessel. i\fter 35 nu'nutes the first drops appeared at the lower 
end. While now in the case of potassium chlorid already tlie first 2 cc. 
showed a moderate antl tlie second 2 cc. a considercdjle reaction for chlorine' 
with silver nitrate, there was no iodine reaction obtained in the first 25 cc. 
of the filtrate. /\fter this a moderate reaction appeared in the next 6 cc. 
and a strong reaction in the following 2 cc. 

Second test: — Here the cohmm of the soil was higher, namely 15 cm., 
but the tliameter was smaller than in tlie former case, namely 3 cm. While 
tlu: weight of the fine soil used in the first case was 200 g-, it was here only 
84 g. The solutions were added in this case in such a manner that the 
surface of the soil was constantly covered b\' it in a height of 2 cm. The 
total (]u.intity of solution adcUd was lOO cc .After about one hour the 
first drop appeared at the lower k\\(\, and while- the chlorine reaction was 
obtained with the first 2 cc tlu-re was no iodine ri-action noticetl in the first 
15 cc y\fter this the next 3 CC. showetl a weak and the following 3 cc a 
strong reaction for iodine. Both tests ])ro\'eil decisixely that an ioilid is 
much better absorbed in the soil than a chlorid. 'i"he calculation shows for 

' .\ foiitrnl Irst was niailc with a distillal waliT iVt'c iVcim ( hlnriiu'. llio lirst few cc. of lliis 
lilUalc sliowcd a weak rcacticm for cliloriiic owiii^ to tlic clilorid alicady iircsciU in tlic soil, l)ut tlic (ur- 
lii<lity was luucli li^Iitcr than in tlic case of potassium chlorid solution. 



Arc Soli!l)Io Iodi«ls AbsoilHMl 1)y tlio Soil ? 52 1 

the first experiment tliat 100 <^. soil absorbed 0.0125 '^- potassium iodid and 
in tlie second case 0.018 14-. The effect depends naturally much on the 
hei"-ht of the soil stratum. 



BULL. AGRIC. COLL. VOL. V. 



PLATE XXIV. 



Fi^J 




Fig II 




Fig. I. Agar-plate from the digestive juice of a silk worm 30 days at room 
temperature. Original plate. 
P'ig. II. The same. Second dilution. 



Bl'LL. AGKIC.COl.l.. rOL. T. 



FI ATK XXV 




Plate showing the stinudating action of rubidium chlorid upon havlov. 
I Ruliidiuni plants. II (\inlrol plants. To page 4(i4. 



S I 




BULL. AC RFC. COLT.. VOL. ]'. 



PLATE xxvrn. 




1 11 III 

Plato shdwiiiii the iniurious ;iclit>n of borax imi barley. 1, 0.05 g borax per Kilo soil. 
IT, o.ol i^ borax. Ill, I'onlrol. To pai^o 51 1. 



'P :^ H it J?> 



^ X *■' * 



^ m m ^ 

^ ^ 3S 



THE 

BULLETIN 



OF TPiE 



COLLEGE OF AGRICULTURE. 



Tokyo Imperial University 



JAPAN. 



Vol. M. 



1904 — 1905. 



CONTENTS OF VOLUME VI 



PACE. 

On the Wax-producing Coccid, P'ricerus pe-la, VVestwood. B}' Prof. C. Sasaki. - i 
On the Feeding of Silkworms with the Leaves of Cudrania triloba, Hance. By 

Prof C. Sasaki. - -_.__. i- 

Corean Race of Silkworms. By Prof C. Sasaki. ---- - 21 

The Beggar Race (Kojikiko) of Silkworms. By Prof. C. Sasaki. ------ zj 

Double Cocoon Race of Silkworms. By Prof C. Sasaki. ' Z'i 

On the feeding of the Silkworms with the Leaves of Wild and Cultivated Mulberry- 
trees. By Prof C. Sasaki. ---------------- ^7 

Some Observations on Anthercea (Bombey) Yamamai, G. M. and the Methods of 

Its Rearing in Japan. By Prof C. Sasaki. -----------45 

-A'New Field-mouse in Japan. By Prof. C. Sasaki. ----------51 

Studies on the Lability ofEnzyms. By K. Aso. --52 

Ueber fungicide Wirkungen von Pilzculturen. Von Y. Kozai und O. Loew. - - 77 
Zur Frage der Existenz des Pyocyanolysins. Von O. Loew und Y. Kozai. - - - 81 

On the Microbes of the Nukamiso. By Sawamura. ----- S'^ 

'Ueber den Kalkgehalt verschiedener tierischer Organe. Von M. Toyonaga. - - 90 
On the Influence of Different Ratios of Lime to Magnesia on the Growth of Rice. 

By K. Aso. --------- y7 

On the Determination of the i\ssimilable Amounts of Lime and Magnesia in Soils. 

By T. Katayama. - ------------- 103 

Ueber den I'-inlluss des Mangans a^if Waldbaume. Von Oscar Loew und Sciroku 

Honda. ----------- -----135 

On the Practical Application of Manganous Chlorid in Rice Culture. I3y K, Aso.- 131 
On the Stimulating Action of Manganese upon Rice, II. By M. Nagaoka. - - - i35 
On the Inlluence of Manganese salts upon Flax. By Y. Fukutome. - - - - - 1 3S 
Can Potassium Bromid Exert any Stimulating Action on Plants? By K. Aso. - - 139 
Can Thorium and Cerium Salts Exert any Stimulating Action on Pha^nogamous 

Plants.? By K. Aso. --------- 144 

Can Salts of Zinc, Cohalt and Nickel in High Dilution Exert a Stimulating Action 

on Agricultural Plants .-* By M. Nakamura. -----------147 

Can Lithium and Caesium Salts Exert any Stimulating Action on Phxnogams .•* 

By M. Nakamura. - ------------153 

On the Siinuil.uing Effect of Iodine and Fluorine Compounds on Agricultural 

Plants. By K. Aso and S. Suzuki. __---! 60 

On the Tieatmcnt of Crops by Stimulating Comp*)unds. By Oscar Loew. - - - 163 
On the Action of Sodium Nitro-prussid upon Plants. By Rana Bahadur. - - - 179 

On the Behavior of Guanidine to Plants. By I. Kawakita. - 1S3 

Physiological Observations on Bacillus Methylicus. By T. Katayama. - - - - 1S5 
On die Occurrence of Bacillus Methylicus II. By T. Katayama .----- ic^i 



PAOF.. 

On the influence of liming upon the action of phosphatic manures. By M. Nagaoka. 1 95 
On the action of various insoluble phosphates upon rice plants. By M. Nagaoka. - 215 
On the effects of soil ignition upon the availal)ility of phosphoric acid for rice 

culture in paddy fields. By IM. Nagaoka. 263 

On organic compounds of phosphoric acid in the soil. By K. Aso. ----- 277 
On the behavior of the rice plant to nitrates and ammonium salts. By M. Nagaoka. 285 
On Different Degrees of Availability of Plant Nutrients. By O. Loew and K. Aso. 335 
On the Injurious Effects of an Excess of Lime Applied to the Soil. By S. Suzuki. 347 
Is the Availability of Phosphoric Acid in Bonedust modified by the Presence of 

Gypsum } By T. Katayama. --- -_-.. -j^^ 

Ueber den Kalkgehalt verschiedener tierischer Organe. By M. Toyonaga. - - - 357 
Ueber das Verhalten von Fluornatrium zu Blut. By M. Toyonaga. - - - - - 361 

On the Flowering of Bamboo. By Oscar Loew. 365 

Further Observations on Oxidases. By K. Aso. 371 

On the Large Bacillus Observed in Flacherie. By S. Sawamura. 375 

Some New Varieties of Mycoderma Yeast. By T. Takahashi. - - - - - - -387 

Can Nitrite Provide Oxygen in Anaerobic Culture for Bacteria ? By T. Takahashi. - 403 
On Manuring with Kainit. By S. Suzuki, ------------- 405 

On the Influence of Various Ratios of Phosphoric Acid to Nitrogen on the Growth 

of Barley. By Rana Bahadur. 421 

Can Aluminium Salts Enhance Plant Growth .? By Y. Yamano. 429 

On the Application of Freezing in the Preparation of Certain Articles of Food. 

By T. Katayama. 433 

Notes on the Detection and Determination of Fusel Oil. T. Takahashi, - - - - 437 
Is Germination Possible in Absense of Air .i* By T. Takahashi. 439 



On the Wax-producing Coccid, Ericsrus p8-la, Westwood. 



BY 



Prof. C. Sasaki, Rigakuhakushi. . 
Agricultural College, Imperial University, Tokyo. Tapan. 



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, 1 that it may also not rarely be found on Ligustrum Japonicum, 
Thunb. (Jap Xedzumimochi). 

Mr. Stanislas Julien^ mentions in his " Xouveaux renseignements sur la 
cire d'arbre et sur les insects que la produisent, etc. PLxtraits 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 I'epoque appelie IMang-Tchong (apres le 5 Juin), ils grimpent aux 
branches de I'arbre, se nourrissent de son sue et laisscnt echapper 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 I'apparence du 
givre, Apres I'epoque appelee Tchouchou (apres le 23 Aout, on I'enleve 
en raclant et on I'appele alors la-tcha, cest-a-dire sediment de cire." 
" Apres I'epoque appelee pe-lou (apres le 7 Septembre), cette cire se trouve 
agglutinee si fortement a I'arbrc qu'il serait fort difficile de I'enlevcr. On 
fait fondre cette matiere, et on la purifie en la passant dans une sorte de 
filtre en etoffe. Ouelques personnes la liquefient a la vapour ct la font 
decouler dans un Vase. Lorsquelle est figee et reunie en masse, elle forme 



2 C. Sasaki. 

ce qu'on appelle la circ d'arbre." " Ouand Ics insectes sont petits (Cest- 
a-dire vienncnt dc naitre) ils sont dc colour blanche. Lorsqu'ils ont produit 
de la cire et qu'ils sont attaint leur vieillesse, lour couleur est ronge et noire. 
Ils se rapprochent entre eux et s'attaclient 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 craisscnt pen a peu et deviennent 
gros comme des oeufs de poulc. Ils sont dc couleur violette et rouge. Ils 
se ticnnent par grappcs et enveloppent les branches ; on dirait c|ue ce sont 
les fruits de I'arbre. 

" Lorsque cet insecte est sur le point de poudrc, il se forme une coque 
(litteralement une maison) qui resemble aux loges des mantes qu'on voit 
sur les Murier. Cette coque s'appelle communement la tch'ong (cire 
graine), ou La-tseu (cire fils). L'interieur est rempM d'ceufs blanc qui 
rescmblent a de petites lentes. On les trouv^c reunis par paquets qui en 
renferment plusicur centaincs. A I'epoque appelee li hia (le 6 de Mai), on 
recueille ces oeufs, on les enveloppe dans les feuilles de gingembre, et on 
les suspend a differentes distances aux branches de I'arbre a cire " 

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 Ilanbury"' 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 arc much 
required, the Chinese statements on the subject being extremely obscure." 

Furtlier Mr. S. Uyeno'' lias published some account of the coccid, chieil}' 
extracted from the Report of the luiglish 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 



On the Wax-producing Coocid, Ericeriis pe-la, Wcstwood. ■s 

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 v.'ax harvested 
in a year, amounts to 600,000 Chin (100 chin = nearly SS'j^eu^). 

For the last four years, I have devoted some time to the study of this 
interesting wax-producing coccid (Ericerus Pe-la Wcstwood), specimens 
of which have been collected by my friends Messrs. M. Shimidzu, Y. ^liyoshi, 
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. i. PI. 
I.). The diametre of the largest specimens measures about 1 1 mm. and 
the height about 9 mm. (Fig. 2. PI. 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. PI. I.). Close to the smaller patches, there opens a very fine 

* One yt-n equals to 2.5 francs nearly. 



A C. Sasaki. 

pore, from which is secreted a very fine Hght 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 
yellov; drop, hangs down from the body of the insect, and finally they drop 
down on the ground (Fig. i, a. PI. 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 enough 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- 
PI. I.). Male coccid : — Cylindrical, somewhat tapering towards the two 
extremeties (Fig. 5. PI. I.). Mead 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 wlien 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 j)airs of smaller roundish ones, lie at the 



On the Wax-prodiicinsr Coccid, Ericerus pe-la, Westwood. e 

sides of the vertex between the dorsal and ventral larger ocelli (Fig. 6. PI. 
I.). The antennae are long and composed of ten segments, and covered 
with long hairs. The segments arc long except the two basal ones, whicli 
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 b\' 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 ]Ma\-, and 
the young larvae begin to hatch out at the bcgining of June. 



6 C. Sasaki. 

The newly hatched larva is light orange yellow, long oval and 
depressed (Fig. 7 and 8. PI. I.). It is about 0.61 mm. in length and 
0.37 mm. in breadth. The body is composed of eleven segments. The 
antennas 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 fevv^ 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 setas, forming an elongated loop on 
the ventral side of the body, lie beneath the epidermis of the larva, 
through which the setae can be seen very distinctly. The thoracic 
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. 
l^etween 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 abtlominal segment. 

The larvae 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: — l^ody oval, depressed, pale greyish brown, 
with a wide indentation at the jiosterior end (Fig. 9. PI. I.). It is about 
0.70 mm. in Kiigth, and 0.42 mm. in breadtli. The dorsal surface of the 



On the Wax-producing Coccid, Ericerus pe-la, Westwood. n 

body s densely covered with snowy white, entangled filaments secreted 
by the dermal glands (Fig. lo, a. PI. 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 
antennae, which are composed ol three segments, arc 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 setre, is wide apart from the remaining two pairs,- 
which lie very close to each other. 

In the last part of .Vugust, the male larxaz (of the 2nd stage) is com- 
pletely imprisoned within an oval cocoon formed by snowy white filaments 
(Fig. 10, b. PI. 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. PI. 11). These white masses of 
cocoons are again covered with white long and flossy filaments. Within the 
cocoons, the larva; undergo the second moulting, and pass to the third 
stage. 

?klale 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. PI. 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. Antennae lie 
ventrally at the front c\-\i\ of the body, and is composed of rine 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 setae 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, lying close to each other, are 
widely separated from the first. 

About the beginning of October, the larvae 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. Antennae, 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 v.'inged insect 
appears tlirough 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. PI. II.), 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 verj^ 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. PI. 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 
1.35 mm. (Fig. 16. PI. II.). The dorsal surface is light greenish yellow, 
with more or less depressed punctuations. The exuvia; lie cxcentrically 
on the dorsal surface in the form of an elongated narrow ridge. The 
postcrir>r end f)f the body is marked with a deep narrow cleft. The caudal 
lobes lying on either side of the cleft are long o\al and crimson red in color. 
The margins of the body an' thickened, and are jirox'ided with a scries 



On the IVax-prodiicing Coceid, Ericprus j)e-la, Wcvtwood. g 

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 bod)', 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 longi- 
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 seta^ are borne on an elevation lying 
at the front portion of the central depression. The antennae (Fig. 18. PI. 
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 ist 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, PI. II.). The first pair of legs, lying at the sides of the anterior 
swelling, where the rostral sctcC 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 \'entral 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 \-entraI surface (Fig. 19. PI. 
II.). The longer diameter of the bod)' is 5 mm., the shorter 3.3 mm. 
and the height 1.32 mm. The dorsal surface of the bod\' is now light 
greyish orange yellow, and is covered all over with dark punctuations as 
well as sparsely scattered short spines (Fig. 19, a. PI. 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 



10 ^'« 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. PI. 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. PI. II.). 

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. PI. I.). Later, the coccid becomes mature as shown 
in the Figs, i and 2. PI. 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 larvae 
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. PI. II.). L^ach two of these sacs are 
tied together by the petioles of the leaves or some other material, and are 
thus transported to some localities \viiere 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.x of the coccid are largely infested by a parasitic chalcid tly, 
apparently of the genus luicyrtis (Fig. 23. PI. 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 breadtli is nearly equal to that 



On the Wax-producing Coccid, Ericcrns, po-la, Wcstirood. 1 1 

of the thorax. Eyes oval, dark reddish brown. OcclH hght yellow, and 
arranged on the occiput widely separated from each other. Antenna; 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 gth, loth and nth segments 
dark brown. The thorax is nearly 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 arc 
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 
2 mm.; the expansion of wings 4 mm. 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, antenna; 
colored uniformly light brown ; the front wings lack the light brownisli 
patches. The male genitalia are pretty long and project bej'ond the end 
of the abdomen. Length of the: body is about 1.2 nun. ; expansion of wings 
about 3 mm. 

As the results of my study on this interesting coccid insect, I 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 ; 



J 2 C. Sasaki. 

Ligastrum Ibota, Sieb., and Fraxinus pubinervis, BI. in Japan, and Fraxinus 
chinensis, Roxb. in China. 

List of References. 

1. R. Ono, Honzokomoku Kcimo Vol. XXXV. P. 9. 18 10. 
Comptes Rendus. Tom. X. P. 619. 1840. 
Loc. Cit. P. 624. t 

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. 188S. (Japanese.) 



On the Wax-producing Coccid, Ericerns pe-Ia, Westwood. j^ 

Explanation of Plate I. 

Aggregation of Female coccid on the branch of Fraxinus pubinervis, Bl.; a, oil drop. i/i. 

Mature female coccid; a, top view; b, side view; c, looked from behind, i/i. 

Dorsal view of ditto. Zeiss B/I. 

Eggs. Zeiss A/I. 

Male coccid. Zeiss aa/I. 
Fig. 6. Head of ditto. Side view ; Fig. 6, a. Top view. Zeiss B/I. 
Fig. 7. Newly hatclied Larva. Dorsal view. Zeiss B/I. 
Fig. 8. Ditto. Ventral view. Zeiss B/I. 

Fig. 9. Larva of the 2nd Stage. Dorsal view. Zeiss B/I. 
Fig. 10. Section of the dermal gland of male larva of 2nd Stage. Fig. 10, a. filaments secreted 

by the gland. Zeiss D/4. 
Fig. II. Femal coccid collected at the beginning of May. i/i. 



Fig. 


I. 


Fig. 


2. 


Fig. 


3- 


Fig. 


4. 


Fig. 


5. 



i^ C. Sasalii : On the Wax-protluciiig- Coccid, Eiieenis, pe-la, West«oo(l. 
Explanation of Plate II. 

Fig. 12. Bi-anch of Fraxinus pubinervis, BI. with the group of cocoons, and with young and 

mature female coccids. i/i. 
Fig. 13. Male Larva of the 3rd Stage. Dorsal view. Zeiss A/I, 
Fig. 14. Ditto. Ventral view. Zeiss A/I. 

Ffg. 14, a. Antenna ; Fig. 14, b. Leg. Highly mag. 
Fig. 15. Winged males getting out of coccons. 2/1. 
Fig. 16. Young female coccid. Dorsal view. Fig. 16, a. Side view. Fig. 16, b. Ventral view. 

Zeiss, aa/i. 
Fig. 17. Long transparent spines along the periphery of ditto. D/L 
Fig. 18. Antenna. Fig. iS, a. Left fore leg. D/I. 
Fig. 19. Female coccid collected in January. Dorsal view. lo/i. Fig. ig, a. Small spines on the 

dorsal surface. Zeiss F/I. 
Fig. 20. Spines on the periphery of ditto. Zeiss D/I. 

Fig. 31. Ventral scale left on the branch after the removal of coccid. lo/I. 
Fig. 22. Triangular sac for transporting mature female coccids. ^. 
Fig. 23. Parasitic chalcid fly (Encyrtis sp.?) 20/1. 



BVH. AGRIC. COLL. VOL. V/. 



FL.iJt: J. 



Fisr H. 




S asaki e( Yokoyama del. 



BULL.AGRIC. COL L. I'OL. V/. 



PLATE II. 




Sasaki et Yokovama 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 igo-i. 
(Witli 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 i. i. PI. 
Ill and IV.) 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 : — 

P'irst stage : The larvoe or silkworms of the Cudrania race"' came forth 
on May 5th at the temperature of 65° — 6^"^ 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 i cm. Moulting 
began on I\Iay 12th, and ended on 13th. 

2nd stage : On ^lay 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 INIay, 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, 
I I have named the Cliinesc race feeding on Cutlrania triloba, Hance " Cudrania race." 



J 5 C. Sasaki. 

and given regularly six times a day. The average length of ten largest 
worms at the end of this age was about 4 cm. :\Ioulting began on the 29th. 

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 -j.-j 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 cxpriments, 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. PI. lY.). 

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. The 3rd 
body segment bears no markings. The markings on the 5th and 8th 
segments arc 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. Tl. IV.). 

The worms of this race, without passing the 4th moulting or casting, 
which is usually tiic case in oilier 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. 



Oil the Feeling orSilk\vorm> with tlis Lsavci of Ciidrauia triloba, Hance. ij 

6. 7. PI. IV.). Their Iciif^th varies from 37 to 43 mm. and their breadth is 1 5 mm. 
on the average. The coloration is of two sorts — white and orange yellow. 

The whitish cocoons (Fig. 5. PI. 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. PI. IV.). 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 an}' 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 

with it and mulberry) 



Awobik 


i race 


7 ' 


lays 


5 


.. 


6 


» 


7 


.. 


S 


•' 


-V- 


dnvs 



I St stage . 
2nd stage. 
3rd stage . 
4th stage . 
5th stage . 



8 day; 

8 „ 

9 " 
7 V 



i8 



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 : — 



Aver, length of ten filaments 

Aver, weight of ditto at the length of 
400 aunes 

Aver, litre of ditto 

Aver, number of duvets of ditto 

Aver, number of ruptures of ditto ... 



Physical nature of the filaments 

reeled from the cocoons of 

silkworms fed ■with 

Cudrania only. 



516 aunes 
(613,04 metre) 

0,147 grms. 

1,96 denier 

i>i 

0,2 



Physical nature of the filaments 
reeled from the cocoons of silk- 
worms fed with Cudrania 
and mulberry. 



(6 


527 
27>i3 


lunes 
metre) 




0,146 


grms. 




1,96 


denier 




o,S 






0,3 





It is very interesting to observe that the parasitic maggot (Larva of 
Ugimyia sericaria^, 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 i)eriod with our 
native races, and the consumption of the meals is also similar to the 
attcr. 

2nd. The quality and quantity of tlic filaments reeled from the cocoons 
of the Cudrania race, are never inferior to our native races. 



J 



i 



On the Feeding of Silkworms with the Leayes of Cudrania triloba, Hance. m 

3rd. If the 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. I. Branch of Cudrania triloba, Hance I '2. 

Plate IV. 

Fig. 2. Largest leaf of Cudrania triloba. Hance i/i. 

Fig. 3. Mature silkworm forming a yellowish cocoon i/i. 

Fig. 4. „ „ ., a whitish cocoon i/i. 

Fig. 5. Wliite cocoon i/i. 

Fig. 6. Yellow cocoon i/i. 

Fig. 7. Fitto of varied coloration. I'l, 



BULL. AGRIC.COLL. VOL. V/. 



PLATE jn. 




BULL AGRIC. COLL. VOL. V/. 



PL A TE ly. 




Fig^y-S'T' Yokoyama del. 



Corean Race of Silkworms. 

BY 

Prof. C. Sasaki, Rigakuhakushi. 

Agricultural College, Imferial University, Tokyo, Ja 

June Tgoj. 
(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 arc laid, are soft nearly rectangular pieces of 
paper of about 24 cm. by 17 cm. The surface of the cartons on which the 
eggs lie, is evenly covered with the ashes of m.ulberry 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 eiven them each da\- are as follows : — 



Number of clays 
taken for breeding. 


Dato. 


Ages. 


Number of meals given 
jier day. 


I 


7 V. 1903 


I. 


.- 


2 


S „ 


M 


7 


3 


9 .. ,. 


" 


7 


4 


10 ., 


•> 


/ 



22 



C. Sasaki. 



Number of day 
taken for breeding. 


Date. 


Ages. 


Number of meals given 
per day. 


5 


II V. 1903 


I. 


7 


6 


12 „ „ 


,, 


7 


7 


13 ,. „ 


» 


I 


8 


14 „ » 


„ 





9 


IS » » 


11. 


2 


lO 


16 „ „ 


„ 


6 


II 


17 » » 


„ 


6 


12 


18 » ,. 


„ 


6 


13 


19 " ,' 


>. 


6 


14 


20 „ 


» 


6 


15 


21 „ „ 


» 


5 


i6 


22 „ 


III. 


*> 
J 


17 


23 „ „ 


» 


5 


i8 


24 „ » 


.. 


5 


19 


25 „ „ 


.. 


5 


20 


26 „ „ 


>• 


5 


21 


27 „ „ 


" 


5 


22 


28 „ „ 


» 





23 


29 „ „ 


" 





24 


30 » .. 


IV. 


4 


25 


31 ,. ., 


., 


4 


26 


I VI. „ 


» 


4 


27 


2 „ „ 


„ 


4 


28 


3 .. .. 


» 


4 


29 


4 ., ,. 


>» 


4 


30 


5 » .. 


» 


4 


31 


6 „ „ 


became r 


nature 




20 ,, 


imago ap 


peared 



From the above tabic, we sec that tlic Corcaii 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 Silkirorms. 23 

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 : — 

I St. Body whitish with a pale bluish shade at the junction of the body 
segments. Head dull greyish brown ; the dorsal shield of the ist 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 nth 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.'j cm. to 6.9 cm. (Fig. i. PI. 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 arc mostly larger and very irregular and 
variable in shape, and they 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 arc nearly oval, pale purplish 
blue ; those on the 8th, and 9th arc 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. PI. V.). 

Cocoons arc either white or light green. The shape is also very 
irregular. Double cocoons arc produced abundantly. 

3rd. Body whitish with light bluish and yellowish shades. Tiie head 



24 (■• Sasaki. 

and the dorsal shield of the ist 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 iith 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 
(Figs. 3. 3, a. n. v.). 

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 ist 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 witli iDJack, 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 i ith and 12th segments with two 
bands. Each of these bands except the ist, 2nd, and 4th, is marked dorsal- 
ly with two jDairs 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 9tli 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 arc light grej'ish yellow. The length of the bod}' 
same as in the ist variety (Figs. 4. 4, a. PI. V.). 

Cocoons are snowy white, and of \ariable shape ; but normally they 
are oval and without a constriction at the mitldle. 

5th. l^ody pale bluisli wliite. Mead and the ilorsal shield of the ist 
setrment colored iiornialK-. The 2nd segment is marked dorsallv with a 



Coreau Race of Silkworms. 25 

dark greyish broad band, which broadens posteriorly. Beside this band, 
there He 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. IMoreover, the dorsal surface of the 4th to nth 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 loth 
segments, there lies, on each, a \ shaped pale bluish marking. The length 
of the body same as in the ist variety (Fig. 5. PI. V.). 

The cocoons of the above mentioned five varieties are equally very 
v'ariable 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. PI. 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 

,, 2nd ,, ., white or light green 

,, 3rd ,, ,, deep yellow 

,, 4th ,, ,, snowy white 

,, 5th ,, ,, white or light green. 

4th variety are much superior in their brilliancy. 
The double cocoons, which amount to about 50 ^q of the total, are 
irregularly roundish or oval, and more or less depressed. Their diameter 



The cocoons of the 



26 



C. Sasaki : Coreaii Bace 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. PI. V.). 

The following gives some of the qualities of the cocoons of the five 
varieties selected from the Corean race. 





ist Var. 


2nd Var. 


3rd Var. 


4th Var. 


5 th Var. 


Aver, length of lo filaments 
(Bave) of cocoons. 


363 Aunes. 
(432 Metre.) 


329 Au. 
(3S2 M.) 


422 Au. 
(503 M.) 


3SS Au. 
(462 M.) 


316 Au. 
(376 M.) 


Aver, titres of ditto at 

400 aune?. 


1.36 Denier. 


1.2SI). 


1.21 D. 


1.43 D- 


1.28 D. 


Aver, duvets of ditto. 


1-5 


2.0 


0.6 


1.9 


2-3 


Aver, ruptures of ditto. 


0.1 


0.2 


0.6 


03 






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. 



Kig. I. Corean race 1st variety i i. 

Fig. 2. iJitto 2nd „ i/i. 

l''g- 3- yyttto 3rd „ i/i. 

Fig. 4. Ditto 4th „ i/i. 

Fig. 5. Ditto 5th „ i/i. 

Figs. 6-1 1. Cocoons of Corean race i/i. 

Figs. 13-15. Double cocoons of ditto i/i. 



BULL. AGRIC. COLL I 'OL . V/. 



PLATE V. 



Fig J. Fig-Z. Fig. 3. Fig J. Fig-J. 'Fig-.e. Fig-. 7 






UT 



fm^ 






Via 



'i' 



V ) 



U- ■-/• 



w 



/\>. (Ir 



— , 1i 






^J> ^^, 






/V^r ■? . a /■ '. 




Fig. 4, a . <l'- 



m 




Fig.n, 



Fig-. IB 



X 



/■ 



Fig-. W. 



Fig. I if . 



Fig. 9. 



Fig. //. 



Fia-. /S. 



L 



The Beggar Race (Kojikiko) of Silkworms 



BY 



Prof. C. Sasaki, Rigaku/iakushi. 
Agricultural College, Imperial University, Tokyo, Japan. 



Align St 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 unknown 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 at a 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. 



Ages 


Dates 


Number of days 
required for cultivation 


Number of meals per 
day 


I. 


3 V. 1903 


I 


3 




4 ,, „ 


2 


4 




5 " " 


3 


4 




6 „ „ 


4 


7 




7 ,. ,> 


5 


5 




8 „ „ 


6 


4 




Q ,. „ 


7 


5 




10 „ „ 


8 


6 




II » „ 


9 


3 




12 „ „ 


10 





n. 


13 " » 


II 


5 




14 „ „ 


12 


4 




15 » » 


13 


4 




16 „ :, 


14 


4 




17 =. ,, 


15 


6 




l5i „ „ 


16 


J 


III. 


19 » „ 


17 


2 




20 ,. „ 


iS 


6 




21 „ „ 


19 


6 




22 „ „ 


20 


6 




23 ,. » 


21 


S 




24 „ M 


22 


5 


iv^ 


25 „ „ 


23 


I 




26 „ „ 


24 


4 




27 .. „ 


25 


5 




28 „ „ 


26 


5 




29 ., » 


27 


5 




30 „ „ 


28 


5 




31 .. .. 


29 


2 


V. 


I VI. ,903 


30 


2 







31 


4 



I 



The Beggar Race (Kojikiko) of Silkworms 



29 



Ages 


Dates 


Number of days 
required for cultivation 


Number of meals per 
days 


V. 


3 ^'- 1903 


32 


4 




4 „ » 


33 


4 




5 „ ,. 


34 


4 




6 „ „ 


35 


4 




7 „ „ 


36 


4 



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. 



Ages 


Dates 


Number of days 
required for cultivation 


Number of meals 
per day 


I. 


4 VII 1903 


I 


3 




5 >. » 


2 


7 




6 ,. „ 


3 


7 




7 „ ., 


4 


7 




s ,, 


5 


7 




9 » » 


6 


9 




10 ,, „ 


7 





II. 


II ,. „ 


8 


6 




12 „ V 


9 


8 




13 ,. .. 


10 


8 




14 „ „ 


" 





III. 


15 „ .. 


12 


6 




16 „ ,. 


13 


6 




17 •, „ 


14 


6 




18 „ „ 


IS 


5 




19 » >. 


16 






C. Sasaki. 



Ages 


Dates 


Number of days 
required for cultivation 


Number of meals 
per day 


IV. 


20 VII 1903 


17 


6 




21 „ „ 


18 


6 




22 „ 


19 


6 




23 ,. „ 


30 


7 




24 ; 


21 


4 


V. 


25 ,. „ 


22 


7 




26 „ 


23 


7 




27 ,. „ 


24 


6 




28 „ „ 


25 


6 




29 ,> 


26 


6 



On the 30th July the silkworms matured and commenced to spin cocoons. 
On the loth 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.'/ cm. in length. 
There are two varieties of these worms, viz : — ist : Body is white, its anterior 
and posterior segments of a faintly yellowish hue. Head dull greyish brown, 
the dorsal shield of the ist 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. i. PL. VI.), 2nd. : The color of the 
head and body as well as that of the last body-segment, of the 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 (Eojikiko) of Silkwoins 3 1 

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. V"I.). 

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. 

Aver, length of 10 filaments 2.95 aunes = 35i metre. 

Aver, titre of 10 filaments for the length of 400 aunes (476 Metres). 

1.35 denier 

Aver, number of duvets of 10 filaments for the length of 400 aunes. 

0.9 

Aver, ruptures of 10 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. 
10.2 denier 

Aver, tenacity of ditto 40.0 grams 

Aver, elasticity 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. 



i 



Double Cocoon Race of Silkworms. 



BY 



Prof, C. SasaM, Rigakuhakushi. 

Agricultural College, Imperial University, Tokyo, Japan. 
August IQOJ. 

(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 tht natives 
with great ease It appears once a )'ear, 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 }'ellowish cocoons. 

ist variety : — Body is light bluish white ; head and the dorsal shield of 
the ist 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 th.e 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 cur\'ed line. The markings on the 
8th segment are round and greyish yellow, the iieripheries are lined with 
black, and a small whitish spot lies at the centre of each. The 5th to iith 
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 j.6 cm. (Fig. 5. PI. \T.). 



■JA t'. Sasaki. 

2nd Variety : — Body light yellowish white, head and the dorsal shield 
of the ist segment colored same as in the ist 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, PI. VI.). 

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, PI. VI.). 

The double cocoons, wiiich 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, PI. VI.). They enclose usually more than two 
chrysahds, and not rarely even seven or eight chrysalids (Fig. 12, PI. 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. 



Double Cocoon Race of Silkworms. 



Explanation of Plate VI. 

Mature larva of beggar race witliout markings i, i. 

Ditto „ „ „ „ with „ i/i. 

Cocoon showing the coloration. 

Showing the sizes of the cocoons of two breeds. 
a, of spring breed. b, of summer breed. 

Mature larva of double cocoon race, ist variety. 
a, dorsal. !', side view. 
Fig. 6. Ditto 2nd variety. 

Figs. 7, 8. Simple cocoons i 'i. 

Figs. 9, lo, II. Different forms of double cocoons I'l. 
Fig. 12. Double cocoon cut open to show the contained pupce i 'i. 



Fig. 


I 


Fig. 


o. 


Fig. 


J- 


Fig. 


4. 


Fig. 


5- 



BULL. AGRIC. COLL. VOL. V/. 



I' LATE VL 



Fig. 6. 



Fig. 5. 



F' 



^r- 




Fig.8. 





Fig. 9 . 



Fig. 10. 



C^ 




Ml ' 



^M^ 



r^ 



\3^^ 



\l 



Sasaki <7 Yokovajiia del. 

/-^. DcuhlL-Cocooii Rccc 
j-i2. Bi'g^er-Ract\ 





'ig. 11. 



^^-*^^- 



On the Feeding of the Silkworms with the Leaves of Wild 
and Cultivated Mulberry trees. 



BY 



Prof. C. Sasaki, Rigakuhakiis/ii. 
Ai^ricultural College, Impcriiil University, Tokyo, Japan. 



In the year 1500, 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 ^^'hether 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. INT. Tokunaga, to wliom 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 arc 
two principal ways, viz. — ncgari (cultivated) and takagi (uncultivated or 
wild). 

I. The ''ncgari'' method is extensivel)' practiced in the north-western 
districts of our main island, where it is esteemed as an improvement. Ac- 
cording to this method, the young shoots prepared from cuttings, arc plantetl 
in a farm in the proportion of 300 to 600 to a single tan.^ 

The young shoots thus planted are left for two }-ears without cutting, 
and in the spring of the third year, the shoots together with the branches, 
are cut off at the height of about 3 to 5 inches above the ground, and the 
leaves are used for feeding. 

In June and July, many \ig(M'ous }-oung branches soon come out from 
the short stock left after cutting. These branches generally grow to the 

1 Tan is equal to 1/4 acre. 



28 C- Sasaki. 

height of sev'eral feet or in favourable conditions over ten feet. In the fourth 
year, the branches left over from the previous year are cut off in May or 
June exactly in the same manner as mentioned before. The mulberry trees 
treated by the negari method, are usually vigorous, and yield an abundant 
crop of fine and large leaves ; but they have always a tendency to be affect- 
ed by the so-called dwarf disease widely distributed in our country. 

2. The takagi method, or that in which the trees are left to their 
natural growth, is at present largely practiced in the north-eastern as well as 
the western districts of our main island. The young shoots are usually pre- 
pared from seeds, layerings, graftings or cuttings ; but those by seeds are. 
mostly employed as stocks for grafting, while the cuttings are only in rare 
cases selected as shoots or stocks for grafting. 

In the spring, some time before the appearance of the leaves, the stems 
may be cut off at the height of about four feet from their base, and planted 
in regular files in a farm. In April or IMay, each stem bears several vigorous 
branches, which are left over to the next or 2nd year without gathering 
leaves. In the second year, before the sprouting of the leaves, the branches 
of the last year, are cut off close to the stem, leaving only three vigorous 
ones, which may also be cut off at the distance of 2—3 feet from the stem, 
according to their lengths. From each of these three branches left, may 
again come out several branchlets, which grow to certain lengths during the 
same year. The gathering of the leaves from the branches or branchlets 
for feeding j)urposes begins in the third year, and thence year after year, 
the same method of gathering leaves is repeated until the stems die from 
age. In still other localities, the young shoots prepared from seeds, graft- 
ings or cuttings &c., are planted in a farm, and are allowed to grow in a 
natural condition, no care being taken as to the arrangement or cutting of 
the branches. It is generally tliought that the mulberry trees cultivated 
after the takagi method, yield generally smaller, rigid, and less nutritious 
leaves than in the negari methotl, whose leaves are larger, more succulent, 
and look to be richer in nutriment. 

It appears to mc that, in l-'rance anil some otlicr l-2uropean silk-rearing 
countries as well as in China, the mulhi-rry trees are treated similarly as in 
our takagi method. 



Feeding of Silkworms with Leaves of Wild and Cultivated Mulberry Trees. 3^ 

In 1898, ]\Ir. F. Lambert/ director of the sericultural station at Mont- 
pellier, after some experiments on the two sorts of mulberry trees, — wild and 
cultivated — concluded as follows : — 

" Dix pour cent environ des educateur font exclusivement usage de la 
feuille sauvage et paraisent s'en bien trouver. Les bons effects de I'alimenta- 
tion avec cette feuille plus fine et plus nourrissante que la feuille greffee, 
poussee dans le meme milieu sontmanifestes. Les educateur faites avec cette 
feuille out rendu en moyenne plus des 57 Kilogrammes de cocons par once, 
tandis que les vers nourris avec de la feuille greffe seulc or associee a la 
sauvageonne ont a peine donne 47 Kilogrammes." 

The above experiment tells us that the silkworms reared with the wild 
mulberry trees yield a larger quantity of cocoons than those reared with the 
cultivated. It seems, however, the mode of their cultivation differs in details 
from ours, which makes us quite unable to compare the results of my experi- 
ment with that of F. Lambert, 

A most renowned weaver, ]\Ir. J. Kawashima of Kyoto, holds the opinion 
that the raw silk produced in the province of OmiXShigakcn) are far superior 
in strength and gloss to that of other districts ; and for this reason it is used 
as a material for winding around the hilt of our swords, and also for making 
the strings of the Japanese guitar (Samisen). The characteristics of this silk 
depends, without doubt, according to I\Ir. J. Kawashima, mainly upon the 
mulberry trees, which arc cultivated after the method of takagi, and not 
after the ncgari method. Till now, we have had no opportunity to examine 
the chemical composition of the leaves of the mulberry trees cultivated by 
these two different methods. 

The main object of this paper is to ascertain whether the silkworms fed 
with the leaves of these two different sorts of mulberry trees afford silk of 
different qualities or not. 

In order to solve the question, I took 5000 individuals of the race Mata- 
mukashi (one of the best white race\ and reared them in May 1900 at the 
temperature of 70-' to "jG Y . Before rearing, they were separated into two 
equal numbers, and reared with the two different sorts of leaves mentioned 
before. 

1 Monitcur des Soics Xo. 1931. 30, IX. iSoO- 



40 



('. Sasalii. 



The number of days in each age or stage of growth, and the average 
weight and length of the worms in relation to the two different food stuffs, 
as well as tlie number of diseased worms during tlie cultivation, are as 
follows : — 

Race Matamukashi. 

The breeding began on the 3rd May and ended on the 6th June, 1900. 



Ages. 


Number of days in 
each a'jjc. 


Aver, weight of 50 full 

grown silkworms at 

each age in 

grms. 


Aver, length of 50 full 

grown silkworms at 

each age in 

mm. 


Number of diseased 
worms at end age. 




7 days 12 lies- 

5 days 10 his 

6 days 21 l>rs 

7 days 9 hrs 


Silkworms 
fed with 
negari. 


Si]k\\orms 
fed with 

takagi. 


Silkworms 
fed with 
negari. 


Silkworms 
fed with 
iiikagi. 


Silkworms 
fed with 
fiegari. 


Silkworms 
fed with 
takagi. 


I. 
11. 

HI. 

IV. 


o,co6 
0,027 

0,153 
0,865 
3,60 


0,0062 
0,029 

0,173 
0,918 
3>7o 


7 
12,5 

24,8 

44,55 
72,6 


7,5 
12,9 

25,4 

46,86 

73.6 


7 
no 

S3 
13 

84 


5 
94 
68 

3 
116 


Y, 


7'l''iys 


Ni 

died 


imber of negari-iu\ worm 
during sj)inning cocoons 


s which 
107 


Xumbc 
hjiinning 


:;r of takagi-iK:Ci. worms 
cocoons 


which died 


during 
200 











The average length often filaments taken out by unwinding the cocoons 
of the worms fed with /^r^^rZ-leavcs was 577,15 metres, while those 
from /c^/vr ^'•/-leaves was 583,10 metres ; the average diameter of the 
filaments f)f the former was 0,0264 mm., while that of the latter was 
0,0192 mm. 

Further, tin; (jualitics of the raw silk recletl from 100 momme' of the 
cocoons jirociu'ed from the- worms fi'd with the two sorts of leaves was as 
follows : — 



' '• Mnmme " i> (.i|ual to 3.75 grms 



Feeding of Silkworms with Leaves of Wild and Cultivated Mulberry Trees. 41 



Raw silk reeled from the 

cocoons of the worms fed 

with «^irrt;7'-leaves. 



Raw silk reeled from the 

cocoons of the worms fed 

with iakas;i-\<i2L\t.s. 



Weight of raw silk. 
Weight of floss silk. 

Tenacity 

Elr.sticity 

Titrc 



11,3 momnic. 
2,76 „ 
47,00 grms. 
11,00 cm. 
12,9 denier-. 



12,7 momme. 
2,25 

48,00 grms. 

11,7 cm. 

I I.I denier?. 



The results of my experiments on feeding the silkworms with two sorts 
of mulberry leaves, viz. wild (A) and cultivated (B), may be summed up as 
follows : — 

1. Silkworms fed with A and B take the same length of time for their 
growth. 

2. Silkworms fed with A are larger in size at each stage than those fed 
with B. 

3. The weight and length of the silkworms fed with A exceed at each 
age those fed with B. 

4. That the number of diseased silkworms fed with A is larger than 
those fed with B, depends chiefly upon the presence of parasitic maggots 
(Larvae ofUgimyia sericaria;, Rond.) which prefer A to B. 

5. The length of the filament procured from the silkworms fed with. A 
exceeds that of B. 

6. Raw silks reeled from the cocoons of the silkworms fetl with A 
are mostly of superior qualities to those fed with B. 



Some Observations on Anthercea (Bombyx) Yamamai, G. M. 
and the Methods of its Rearing in Japan. 



BY 



Prof. C. Sasaki, Rigakuhakushi. 

Agricultural College, Imperial Univ-ersity, Tokyo, T;ipaii. 
JlDlC ipOJ. 

WITH PLATE VII & VIII. 



This well-known moth lias been studied by many, both natives and 
Europeans, and a number of works on this subject have been published both 
in the Japanese and forcic^n languages. The principal works are those of 
C. Personnct', Y. Saiki', T. Wardle'', L. Sonthonnax^ S. Yaguchi^, A. 
Wallace'', &c. With regard to the larva at each stage, tlie coloration of 
the moth as well as our methods of cultivation, our knowledge remains still 
imperfect. This leads me to state some of my obser\'ations made on the 
subject during the past years. 

The moth is widely distributed in our countr)', and we may find it in 
almost all the mountainous districts. The principal food plants preferred 
by its larva are Ouercus serrata, Thunb., O. glandulifera, BL, O. glauca, 
Thunb. forma serica, O. phyllireoides, A. Gray and other Onercus species. 

The larva come forth from the latter part of April, and after the final 
moult, they attain full size from the end of June to the beginning of July, 
hence the larval life up to spinning the cocoon lasts for from sixty to 
seventy days. The larva, after being imprisoned in the cocoon, changes 
into the pupa at the end of a week or more, and the moth comes forth 
generally at the end of 40 to Co da}'s after pupation. The single moth lays 
from 150 to 300 eggs, and dies at the end of about a week after its appcar- 

1 C. Fcrsonnct, Lc ^'er a soic du Chenc, 1S6S. 

2 T. Wardle, The Wild Silks of Iiulia, iSSi. 

•' V. Saiki, The Culture of Aiitlir. Yamamai, G. M. 1S7S (Japanese). 

^ Laboratoire d'otudes de la soie Lyon — Essai de classitication des I.epidoptercs productcuis dc 
soie 1897-1S98. 

5 Journal of the Silk Industry Nos. 24, 25, 26, 1S95 (Japanese). 

*• A. \Vallaco, On the oak fecdini; silkworm from T-ip^n. Trans. I'.nt. Soc. Vol. \'. 1S62 — 64. 



44 



C. Sasaki. 



ance. The cg^gs thus laid, hybernate and hatch out in the following spring. 

The eggs are nearly roundish, more or less flattened, and are of a dark 
greyish brown color. The diameter is 3 mm. on the average. They arc 
aid singly or in groups on the stems or branches of the food plants. 

Newly hatched larva-Stage I. Length 9 mm. Head dull ochre brown, 
body light brownish yellow, prothoracic shield light greyish brown bearing 
a pair of small warts each with 4 yellowish hairs. The five blackish 
longitudinal streaks (dorsal, subdorsal, and infraspiracular lines) beginning 
at the 2nd segment of the body, extend as far as the nth segment, and the 
dorsal line is much broader than the rest. In the space between the dorsal 
and subdorsal, the subdorsal and infraspiracular, and the infraspiracular and 
basal lines, there lies on either side of each of the three thoracic and nine 
abdominal segments, a single wart provided with more than five blackish or 
greyish hairs. The subdorsal warts on the 3rd thoracic and the 8th 
abdominal segments are larger and blackish, and those of the latter lie close 
to each other. The suranal plate is marked with a blackish triangular, and 
the outer side of the anal segment (9th abdom.inal segment) with a blackisli 
marking (Figs. I, I, a. PI. VII.). 

luid of stage I. Length 13 mn:. The head prothoracic shield and 
warts on the body-segments colored same as in the previous stage ; but the 
body has now changed into bluish green, and the five longitudinal streaks 
arc deep blue (Fig. 2. PI. VII.). 

Stage II. Length 30 mm. Plead dull ochre brown as in the previous 
stage. Body yellowish green. The five longitudinal blackish streaks have 
disappeared ; but there still remains a bluish dorsal line, and there is in 
addition a pale yellow supraspiracular line. The warts on the body have 
still the same position and are of the same number, they are now colored 
orange yellow and are provitled each with two sorts of black and yellowish 
hairs of various length. The warts on the 8th abdominal segment are con- 
solidated into one. 'Plie blackish marking on the suranal plate is absent, 
while that on each side of the anal segment [Persists (Figs. 3, 3, a. PI. VII.). 

Stage in. Length 42 mm. Head greenish brown. Pod)', dorsally 
above the light yellowish supraspiracular lim-, light }'elk)wish green, and 
vcntrall)' below tin- same line, lively green, and covered s[iarsely with club 



Seme Observations on Anthcnra (Bombyx) Yamaniai, (J.M. 



45 



shaped light yellowish liair. The supraspiracular line, which begins at 
the ist abdominal segment extends to the last segment. The prothoracal 
shield light greenish yellow. The warts on the subdorsal and supra- 
spiracular lines are reddish brown and are provided with white and blackish 
hairs of various length ; those on the infraspiracular lines are bluish and 
bear some blackish hairs. Close to the base of the warts on the subdorsal 
lines of the 3rd thoracic and the ist and 2nd abdominal segments, as well 
as on the supraspiracular lines of the 2nd and 3rd abdominal segments, there 
is a single silvery dot ; but it is often absent on the latter lines. The 
pectoral legs dark brown ; the abdominal legs deep green. The suranal 
plate is bordered at its free edges with a broad brownish marking, which 
encloses a dark bluish patch. The edges of the anal legs are also colored 
brown (Fig. 4. PI. VII.). 

Stage IV. Length 58 mm. Head light green, but its color deepens 
later. The body, dorsally light green, ventrally deep green. ^\ yellowish 
supraspiracular line which begins on the ist abdominal segment, and ex- 
tends to the last abdominal segment, is bordered above with a dark brown- 
ish streak, which meets posteriorly with a broad brownish marking on the 
suranal plate. The warts on the subdorsal, supra, and infraspiracular 
lines are much reduced in size and are colored blue. They bear brownish 
or blackish hairs of various lengths. The number of the silvery dots lying 
on the subdorsal and supraspiracular lines are not definite, and vary in dif- 
ferent individuals. The color of the pectoral and abdominal legs are similar 
as in the previous stage. The following shows the number of the segments 
which bear silvery spots on the subdorsal antl supraspiracular lines. 



Nos. of indivi(.lu;i!s. 


Ncs. of segments bearing; silvery 
dots on the subdoi-sal line. 


Nos. of segments bearing silvery dot 
on the supraspiracular line. 


I 


4 


5 


2 


4, 5> 6 


5. 6. 7 


3 


4. 5- (■> 


5,6 


4 


4, 5. ^'> 


5- 6 


5 


4.5,6 


5.6 


6 


4. 5 


5. 6 



46 



('. Sasaki. 



Nos. of individuals. 



Nos. of segments bearing silvery 
dots on the subdorsal line. 



8 
9 

lO 

II 

12 
13 
14 
15 
16 

17 
18 

19 



4, 5 
o 

4. 5, 6, 7, S, 9, 10 

4, 5- 6, 7, 8, 9 

4, 5, 6, 7, 8, 9 

4, 5. 6 

4, 5, 6, 7. 8 

o 

4 

4, 9, 6, 7, 8 

4, 5. 6 

4, 5. 6 

4, 5, 6, 7 

4, 5, 6 



Xos. of segments bearing silvery dots 
on the supraspiraculars line. 



5. 


6 















5. 


6 






5, 


6, 


7' 


8 


5, 


6, 


7, 


8 


5. 


6, 


7 




5, 


6 






5> 


6, 


7 













5. 


6 






5. 


6 






5, 


6 






5. 


6 






5, 


6 







Silvery dots on the subdorsal lines are usually much lartjer and more 
conspicuous on the 5th and 6th segments than on the rest, and those on 
the supraspiracular lines are generally smaller than the former (Fig. 5. 

ri. VII.). 

Stage V. Length 95 mm. Head deep emerald green. The pro- 
thoracic shield light yellowish green. Body dorsally above the supra- 
spiracular line, light yellowish green, while vcntrally below the same line 
deep green, and covered all over with short yellowish hairs. The supra- 
spiracular lines as well as the dark brownish markings on the last abdominal 
segment and the anal legs are same as in the previous stage. All the warts 
on the subdorsal, spra. and infraspiracular lines arc small, bluish, and pro- 
vided with greyish or yellowish hairs of different lengths, while those on the 
subdorsal lines on the 2nd and 3rd segments are yellow. The silvery dots 
on the subdorsal lines are small roundish and lie close to the warts mostly on 
the 4th, 5th, 6th, and 7th, segments; while those on the supraspiracular lines 
lie mostly on the 5th, 6th, 7th and 8th segments, of which those on the 5th, 
and 6th are much larger and more conspicuous than the rest. The spiracles 



Some Observations on Antlicrwa (Bombyx) Ynmaniai, G.M. 47 

dull brown ; the pectoral Ictjs light brown, while the abdominal deep 
green (Fig. 6. PI. VII.). 

Cocoons : — Length 50 mm., breadth 25 mm. on the average. They are 
oval and compact in texture. The color varies from light greenish yellow 
to deep green. The surface of the cocoon is rather rough and marked with 
irregular fine wrinkles. lit one end of the cocoon, is a single long silky 
pedicel with which the cocoon is tightly attached to the branches, from 
which it hangs down (Fig. 7. PI. VII.). The cocoon is, in this position, usual- 
ly protected by leaves nearly on all sides except one. The protected sur- 
face of the cocoon is usually light yellowish green, while the exposed surface 
is deep green, and resembles closely the leaves which protect it. 

Pupa. — Large, oval, blackish brown. Length 43 mm. Breadth iS 
mm. 

Imago ^. — Head and body bright yellow. Eyes blackish. Antennoe 
plumose with short branches. Prothoracic collar greyish brown. Thorax 
with a tuft of hairs on each side. Wings are always bright yellow. The 
costa of the fore wing is greyish brown and forms a continuous band with 
the prothoracic collar. Fore wing larger than the hind, and with a faint 
orange line running about the middle from the costa towards the inner 
margin. At the middle of tliis line, lies a large transparent eye spot, whose 
inner side is bordered successively from the inner towards the outer by 
reddish, brown, white and reddish arcs, and the outer, by yellowish and 
blackish arcs. A blackish transverse line running between the eye spot and 
the outer margin of the wing is decorated along its outer side with a whitish 
line, and then with a reddish brown coloration. 

On the area lying between the c}'e spots and humeral angle, there lies 
transversely an incomplete zigzag reddish brown line, whose inner side is 
margined with white. 

The eye spot on the hind wing is smaller than that on the fore, and 
surrounded successively from inner towards the outer by yellow and reddish 
brown rings. The outer side of the reddish brown ring is decorated by two 
colored arcs— yellow and black. The upper end of the blackish arc 
broadens into a large oval dot, wr.ile the inner side of the reddish brown 
ring is also decorated by two colored arcs — whitish and reddish brown. The 



48 <^« Sasak?, 

colored streaks lying between the eye spot and the outer margin and bet- 
ween the former and the insertion of the wing are almost sim.ilar to those of 
the fore wings. Length 37 mm. Expanse of wings 130 mm. (Fig. 8- PI. VII.). 

Imago ^. — The coloration is very variable, but there are no specimens, 
which take the same coloration with the female. The branches of the 
antenna are very long. The most prevalent colors of the male are of two 
sorts : — I St. Body greyish brown. Prothoracic collar greyish brown, with 
the front half white. The fore and hind wings are equally colored greenish 
brown with light colored inner areas. The number and position of the eye 
spots and streaks are like those of the female, but their colors change with 
ground color of the wing. 2nd : — Body and wings dull reddish brown. The 
prothoracic collar dull brownish white. The fore and hind wings dark 
reddish brown with lighter colored inner areas. The number and position 
of the eye spots and streaks same as in the ist; but their coloration varies 
more or less according to the ground color of the wing. Length 34 mm. 
P^xpanse of wings 127 mm. (P^igs. 9, 10, PI. VHP). 

Methods of culture. The culture of the larva is only practiced by the 
people in the village Ariakcmura in Naganoken on the open grounds, where 
the food plants are regularly cultivated. The plants selected as the food of 
the worms are of two species — Quercus serrata, Thunb. and O. glandulifera, 
Bl. They are planted on the ground in the proportion of 2 in every three 
tsubo.' The height of the stems as well as the branches are not allowed to 
exceed four feet, as otherwise it would be very troublesome and incon- 
venient in treating the worms. 

In spring about a week before the coming forth of the worms, the eggs 
are pasted on a long ant! narrow piece of thick and stout paper, and the 
latter is tied up on the JM-anclu s of tlie food i)lants. When the larwa come 
forth, they crawl on tf)\vards the young branches and devour the young 
tender leaves. Thus the people allow the worms to grow freely in a natural 
condition. No furtlier care is taken about them, except that the larva are 
at all times protected from birds, tree frogs, wasps, spiders, S:c. If the 
leaves of a trc:e are entirely e.itcn up b)- the worms, its branches are cut off 
ar.d transferred to leaf-bearing trees. 

' iMiho is equal to 6 feet siiiinre. 



Sonio Observjitioiis ou Anthercjpa (IJombyx) Yamamai, G.M. aq 

If the worms attain full growth, they bind together two or more leaves 
by means of threads, within which they spin a cocoon, so the cocoon is 
usually protected by the leaves nearly on all sides with only a small un- 
covered portion. The cocoon is usually colored yellowish green ; but its 
exposed portion is lively deep green. After a week or more, when the ex- 
posed surface of the cocoon has the appearance of being covered with a 
thin white layer (this is the indication of fuiished pupation), the people 
collect the cocoons together with the branches on which they are attached, 
and then hang down the branches on the strings stretclied out horizontally 
under a projecting roof 

When the moths come forth, they are then transferred into a large open 
worked bamboo basket (diameter about half a metre, lieight little less than 
the diameter), within which they are allowed to pair. Each pair is now- 
taken out of the basket, and again transferred to another bell shaped 
bamboo basket (diameter 20 cm., height 17 cm.) with its wide mouth closed 
with a large sheet of paper in order to prevent escape. After a while, the 
female moth will lay the eggs on the inside of the basket successively in 
two or more days. When egg-laying is finished, the moth is removed, and 
6 or 7 empty baskets bearing the eggs on the inside are piled up one upon 
the other, and may be hung down by means of strings under a projecting 
roof mentioned before. Later the eggs are scratched off from the basket 
by the aid of a long piece of bamboo, and then they are spread over on a 
rectangular wooden frame with a bottom made up of strong grass cloths. 

The frame is kept by hanging down in a cool chamber till the follow- 
ing spring, until the eggs can be pasted on the long pieces of paper in order 
to bind them up around the branches of the food plants as stated before. 



50 



C. Sasaki : Some Observation on Antheroea Yanianiai, U.M. 



Plate VII. 

Fig. I. l-aiva of Anthercea Yamamai, G.M. ist stage 5/1. 

Fig. I, a. Abdominal segment of ditto. 

Fig. 2. Larva of A. Yamamai, G.IM, at the end of ist stage 5/1. 

Fig. 3. Ditto 2nd stage 4/1. 

F'g. 3j 2- Abdominal segment of ditto. 

Fig, 4. Larva of A. Yamamai, G.M. 3rd stage i/i. 

Fig. 5. Ditto 4th stage i/i. 

Fig. 6. Ditto 5th stage i/i. 

Fig. 7. Cocoon of A, Yamamai, G.M. i/i. 

Plate VIII. 

Fig. 8. Anthercea Yamamai, G.M. Female i/i. 
Eig. 9. Ditto male 1/2. 

Fig. 10 Ditto male l/l. 



\ 



BULL. AGA'/C. COL J.. I'Of.. 17. 



PLATE VLf. 



Fig. 5. 




Fig.3. 



fig.4. 





Fig. 2. 



Fig.l. 




Fig.3,a. 



Fig. 1/1. 



Fig.7. 




Fig.6. 




^.^ 



^% 



Yokoyaina del. 



BULL. AGKIC. COLL. J'Of. If. 



PL ALE VILL 




Yokoyama del. 



A New Field-mouse in Japan. 

PA' 
Prof. C, Sasaki, Rigakuhakushi. 

Agricultural College, Imperial University, Tokyo, Japan. 

May igoj. 
(With Plate IX.) 



Four years ago, field-mice made their appearance in the prefecture 
of Ibaraki lying north-east from Tokyo, and the injuries to crops extended 
not only over the whole prefecture, but also into the neighbouring provinces. 
In 1900, Dr. S. Onugi' and Mr. K. Sanui made some observations on their 
habits and have carried on their experiments for annihilating them by the 
inoculation of the so-called mouse typhusbacillus. In January 1901, Dr. S. 
Onugi- from a study of the characters of the mice and pointing out the dif- 
ferences between the latter and the house-rat, has concluded that the mice 
was probably of the same species with Arvicola subterraneous, Selys. In 
February of the same year, I also visited the above stated prefecture for the 
purpose of studying the characters and habits of the mice and made public 
my results in the Journal of the Agricultural Society of Japan, No. 235. 

In 1902, Prof Y. Kozai^ after two years' experiments for killing the 
mice by means of Mereskowsky's Bacillus, has obtained a satisfactory result, 
and at present it is practically employed largely in various provinces. 

In the following lines, I will state the results of my studies upon the 
characters and habits of the hateful mice. 

Characters : Body rather small, but pknnp. Winter pelage : dorsalh- 
rusty greyish brown, ventrally greyish white (the dorsal hairs are greyish 
with rusty yellowish ends, while the ventral are greyish with whitish ends). 
There is no marked line laterally between the dorsal and ventral colora- 
tions. Tail rather short, covered sparsely with hairs, its dorsal hairs dark 
grey, and the ventral greyish white. Limbs light greyish brown. Snout 

1 Journal of the Agricultural society of Japan, No. 224, 1900. 

2 Ditto „ ,, „ ,, ., „ No. 233, 1901. 

' The Bulletin of the College of Agriculture, Tokyo, Imp. Univ. \"ol. IV. No. 5. 



e2 <^'« Sasaki. 

rather blunt; eyes dark greyish brown ; the upper incisors not projecting 
beyond the snout. Ears nearly quadrangular (length 13 mm. breadths 
mm.), about 7^ as long as the head, with a few hairs on the inner side, and 
their outer side as well as the free edges are covered with greyish hairs. 
They are not completely concealed within the hairs of the head. Incisors 
yellowish brown on the outer side (Figs, i ; I, a. PI. IX. 1. Hip glands 
are large and oval. Plantar tubercles 5 (Figs. 3 ; 3, a. PI. IX. 1. MammtK 
8, inguinal 2—2; pectoral 2 — 2. 

Skull long, smooth and flattened ; auditory bulla comparatively large, 
bulky, and oval in shape (Fig. 2. PI. 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 
open base ; 3rd. with 3 closed triangles with an anterior and a posterior 
loop, with 4 inner and 3 outer salient angles ; of the lower jaw, 1st. 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. PI. IX.). 

In general aspects and characters, the present species resembles to a 
certain degree Arvicola subterraneous, Selys., which is described by Mrs. PI. 
Leunis' and J. R. Hos.-; 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 feet! the mice in an 
enclosure, they remain (juiet 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 

I II. Ixiuiiis, Synopsis dcr Tlucikuiidc Hand I. 
- K. Bos, Ticrischc Schiidlinge u. Niitzbngc. 
■* Hatanedzumi (Jap.) means Farm mouse. 



A \ew Ficld-iiioiisc in Japan. 5:5 

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 cat them on the spot, or else they carry them to their nests; 
thus tlie 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. 
ri. IX.). 

The nests (Fig. 6. PI. IX.) 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 \'arious plants. The inner 
layer of the nest consists of a mucli 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, Ga^rtn., Daucus carota, L., the stalks or leaves 
of the tobacco plant, ears of the rice plant and others. The roots arc 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 arc equally 
cut off to the length of four to five inches, and then they are piled up 



CA 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. 



A New Field-monse in Japan. cc 

Explanation of Plate IX. 

(Figs, i; I, a, 6. drawn by K. Yokoyama) 

Fig. I. Arvicola liatanedzumi, Sasaki i/i. 

Fig. I , a. Ditto, 

Fig. 2. Skull of ditto ^ i/i. 

Fig. 3. Right fore foot i/i. 

Fig. 3, a. Right hind foot i/i. 

Fig. 4. Molar series of right upper jaw 4/1. 

Fig. 4, a. Ditto of left lower jaw 4/1. 

Fig. 5. Root of mulberry tree with the trace of teeth ; 

a blackish line shows the level of ground. 
Fig. 6. Nest with two openings 1/3. 



BULL. AGRLC. COLL. Vol.. VL. 



I'L.A'I L LX. 



Fi^. I. a 




Fic^. V ^ 



Studies on the Lability of Enzyms. 

BY 
K. Aso. 



The cause of the chemical powers of enzyms has frequently been the 
object of speculation. According to the tlieory 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 E. 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 i 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 t^roupings we must select such 
compounds that have quite specific actions, even in liigh dilution and in 
perfect neutral solution. Loeiv suspectetl formerly that the lability of 
enzyms is caused by the simultaneous presence ot amido — and aldehyde 
groups, but his own tests with alkaline silver solution Hiiled.- The 
presence of aldehydegroui)s would i)rovide a plausible \"iew, as Vcrnon^^ 



1 riliig. .\rcli. 27., 212 ; Die chomisclio Kiiergic dor Icbcndcn Zollcn, p. 149; Jouni. f. prakt. Chein. 
iSSS, p. 194. 

2 rtluj^crs Archiv. fiir die ges. Pliysiologic, vol. 27, p. 212. 

3 Journ. of Physiology, vol, 29, p, 331 [1903]. 



58 K. Aso. 

has pointed out': "it may, for instance by alternate hydration into 
CH (0H)2 groups and subsequent dehyrationbe able to eftect the hydrolysis 
of proteids, whilst ili-amido-or other aldehyde groups by the reverse 
process may be able to eftect the dehydration of caseinogen into casein." 
As to the action of zymogens, Vernon expresses himself as follows : 
•' Let us also provisionally accept Loeiu'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. Loezv, the 
zymogens contain ketonegroups, and that the activation process consists 
in the opening of lactamgroups in the zymogen molecule, labil amido- 
groups thereby being generated.' Amidoketoncs also are very labil bodies 
as seen from the behavior of diamidoacetone which spontaneously changes 
to an indifferent substance {Riigheimer and ]\IiescJiel). In regard to the 
amidogroups O. Loew infers their presence from his observation that 
dilute formaldehyde easily destroys the action of cnzyms at the ordinary 
temperature.- It is well known that formaldehyde easily attacks amido- 
groups of a certain lability, e.g., : 

C,M..NH3 + CH30 = CgH5.N = CH2 -f-H^O 

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 
amidog'roup as such has disappeared. The following table shows the more 
or less intense action of formaldehyde on enzyms. 



* Cciilralljl. f. IJaktcriologic 12, p. 445. 

2 Journal fiir prakt. Chcmic 188S. vol. 37. p. 104. Sucli oliscrvations were later on made also l)y 
various authors. 



studios on tlic Lability of Enzyms. 



59 



Enzyin. 


Formaldehyde.^ 


Time in which 
the enzym is killed. 


Author. 


Diastase 


^% 


in 24 hours 


Bokorny. 


Myrosin 


s% 


soon 


») 


Rennet 


o.^X 


" 


M 


Zymase 


0.05X 


»> 


Wroblewsky 


Sucrase 


s% 


one hour at 54° C. 


Pottevin 


Catalase 


A% 


I hour 


Loew 


Pepsin 


s% 


24 hours 


>) 


Pepsin 


\% 


24 hours 


Sawamura 


Papain 


OA% 


Soon at 40^^ 


Vines 



It is true that formaldehyde will act also on hydroxylgroups and 
produce mcthylene-compounds as, c.l;-., with the polyv'alent 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 arc far different from those just mentioned. The inference that 
amidogroups of a certain lability are concerned in the activity of enzyms 
would receive fiu'ther support, if it could be shown, that the enzx'ms take 



' "I'he coinniorci.U Innniiliii or formal lonlains ahaut 40%' foniialclt'liYilc. 



6c K. Aso. 

up free cyanogen and would lose thereby their activity. ^ As to the 

different labiHty of amidogroups Lociv expresses himself as follows: "Die 

Amidogruppe kann unter gewissen UmstLinden stabil, reactionsunfiihig, 

untcr andcren aber wieder iiusscrst labil und reaktionsfahig sein. Die 

Amidogruppe ist Z. B. im Urcthan schr stabil, im Harnstoff labiler, noch 

mchr im Guanidin. Im Hydroxylamin und Diamid aber ist sic so 

energisch geworden, dass diese Stoffe selbst bei grosser Verdiinnung 

noch in alles Icbende Protoplasma ohne Ausnahme eingreifen konnen, 

d. h. Gifte allgemcincn Charakters sind. Folgende Formcln lassen die 

Einfliisse bcnachbarter Gruppen auf die Energie der Amidogruppe 

erkennen : " 

/NH. /NH„ /NH, 

CO CO C=NH 

"^O.C.H, ^NH., ^NH, 

Urethan Harnstoff Guanidin 

NH, NH, 

I " I ' 

Nil 2 OH 

Diamid (Hydrazin) Hydroxylamin 

When dicyanogen acts on amines, it forms as chief compounds addition 
])ro(lucts with two molecules of amine. Thus 

2(C,II..NH2) + (CN)„ = C,H,-N = C-C = N-C,H,2 

I 1 
lUN NH, 

Wlien, however, it acts on amidocompounds several products ma)' result. 

'Ihus the chief product \\\\.\\ amidobenzoic acid is cyancarbimidamidoben- 

zoic acid CN-C-NH.C.H^.COOH. 

II 
NH 

' Also liydroxylgioups can take up cyanogen at the ordinary temperature as Lonu has shown witn 
pyrogallol. l!ul tliis is the only case known thus far. (Journ. f. prakt. Chcm. Vol. XV. 326.) Very 
lahil methylene groupings as in the acetylacctic ether also can react with cyanogen hut only in presence 
(if sodium ethylate ('^H-ONa (W. Trauhe). Thus far, however, only a few such cases have hcei: 
described. 

'>■ This lormula has heen sliow n liy 7'i<'iii..iiin\n correspond lietler to the behavior of the cyananiline 
tiian the fnrmer iniiddfnrniula. 



studios on the Lability of Enzyiiis. 6 1 

It is noticeable, liowever, that not every amido compound and amide 
can combine with dicyanoc^en, and imidojTroupinc^s are not acted upon at 
all. Neither liydrazobenzol nor asparagine nor urea are acted upon. But 
AndrcascJi has shown that methylthiourea enters into reaction. The 
peculiar behavior of free cyanogen towards higlily labil amidogroupings 
have induced Loeiv and Tsukainoto^ to test the beha\-ior of a highly 
diluted aqueous solution of dicyanogcn 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 dicyanogen 
would act on enzyms when applied in excess to a concentrated solution, 
since Locic has shown that cyanogen combines with ordinary albumin. - 
He applied solutions containing lo— 25^q albumin. In m\' 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, lience a reaction under this condition would admit a safer 
conclusion as to the degree of lability. 

Experiment wit/i Pepsin. 

* grams of commercial pepsiiv' were dissolved in 200 c.c. water and 
divided into halves.^ Cyanogcngas 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 2^0 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 l'"orsclmngsbcn'chtc iiber I.chensmiUcl, \'ol. T. Xo. 7; — Journ. College of Science, Tokvo, 1S06. 
Cf. .also These Bulletins \'oI. II. No. 7. 

2 Journ. I", prakt. Chcni. 1S77. 

3 The solution of this sample was acid. 

* Sonic ether was added to the control llask to prevent bacterial orowtli. 



62 K. Aso. 

and neither the precipitation with nitric acid nor the saturation with 
ammonium sulfate did show any difference ; nor the colorimctric comparison 
of the biuret reaction. In a second experiment, 2£^ 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, wdiile the other half was treated with 
thc same quantity of cyanogengas as before, but in this case, the solution 
was kept at 35-4o''C during the treatment. Moreover this liquid ^was 
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 w^ere 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- 
ment the conditions were again changed. Pepsin, ig, was dissolved in 
200 c.c. water and ig 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 2_^q 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. 
Gi'ccn reports that pepsin is injured by o.o02^q soda solution after 1-2 
hours at bodil)' 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 trom 
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 cither 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 Latighy and Eves found a distinclly inliiliilDiy actiiJii li> lie manilVstcd hy the presence of a.s 
little as 0.0015% of soiliuni carlioiiate. 



studies on the Lability of Eii/yms. 63 

Experiment luith Trypsin. 

I gram of commercial trypsin was dissolved in 200 c.c. of water 
containing- 0.4$^. sodium carbonate and divided into halves, one servinj:^ 
as control and the other being treated with cyanogen gas developed 
from 5 grms 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 zuith Emulsin. 

A solution of o.\% emulsin with o.\% NaoCO., was treated with 
cyanogen gas developed from 5 grms mercuric cyanid. After standing 
for 48 hours, O.ig. 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 fornKition of 
the decomposition products of amygdalin, \'i?:. of glucose as well as of 
benzaldehyde. 

Experiment tvitJi Talcadiastase. 

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 developetl 
from 5 grms mercuric cj'anid 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 O. iJ?q 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. 1 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. 

45b- '^^ '^ fresh radish root were triturated with addition of lOO 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 1 5 hours, the cyanogen gas was replaced by air and the 
liquid tested for oxidizing enzynis 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 acjueous 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 40-5o"C to remove the [)russic acid, whereupon the guaiac and the 
guaiacol test for peroxidase were readily obtained, only the guaiac test 
for oxidase was somewhat weaker. 

In all the cases here described dicyanogen has failed to destroy the 
activity of the enrjynis, what reveals a great chemical difference between 
the lability of enzyms and the lability of the active proteins in the living 
protoplasm. 

Locw and Tsukamoto (1. c.) have observed that a fresh solution of 
dicyanogen in water in a dilution of 1 : 5000 kills bacteria and of i : lOOOO 

1 yVllhouyh tliis solution liad a weak alkaline reaction, the diastase was not perceptively injured. 
Cldltctuicn and also Grulziicr have observed an injurious action of small quantities of alkalies, but 
/■'.pstcui and Siliulzc found lliat herel)y tliis en/.yni is not destroyed, but merely " iiaralyscd," because 
by neutralization the activity is again restored, at least partially. 



studies oil the Lability of Euzynis. 6$ 

phacnogams, 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 
a 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, i 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 : 

(x)NH,+Nr^ r=(x)-X = X-OH-f-H„0 and 

\0H 

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 

Fxpcrinicnt icith Pepsin. 

i). 0.3g of pepsin were dissolved in 300 c.c. water, and dividetl 
into three equal parts. To one potassium nitrite and sulphuric acid in 



* In this connection it is also an interesting lact tliat a very high ihlutcxl nitrous acid (i : looooo) is 
much more poisonous for lower orgam'sms than nitric acid, as Loc~:j anil Bi}korii\' have shown. 



66 



K. Aso. 



high dikition were added in the calculated proportions to produce free 
nitrous acid of O.i^^ in the liquid. To another potassium nitrate and 
sulphuric acid were added in such proportion as to produce o.\% 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 &%^^ white and 10 c.c. of a 1% hydrochloric acid were added. 



After 1/2 hour at 40X. 



After I hour at 4o"C. 



Control. 



Fibrin dissolved ; c^^'g 
white not yet. 



Egg white wholly 
dissolved. 



Nitrous acid. 



Fibrin dissolved ; egg- 
white almost unchang- 
ed. 



Some 'i^g white still 
remained. 



Nitrous acid. 



Some pieces of fibrin 
dissolved ; o.^^ white 
remained intact. 



Egg white not 
attacked. 



An equal quantity of fibrin was again added ; 




After i hour. 


After I hour. 


After 17 hours. 


After 3 days. 


Control. 


Fibrin and egg 
white dissolved. 














'I'rcated with 


Fibrin as well as 
egg white attacked. 


Both almost 
dissolved. 






nitric acid. 






Treated with 
nitrous acid. 


Fibrin and egg 
wln'tc not dissolved. 


Both 
unchanged. 


Fibrin dissolved, 
but not egg white. 


Egg white a little 
attacked. 



2). In a second experiment o.2)'y nitric acid and nitrous acid were 
applied in the same way as before. In the control solution however, 
0.2%' 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. 



StiKlies on llie Lability of Enzyms. 



^7 



Hereupon lo c.c. of a \% 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. 





jVfter i hour. 


After \ hour. 


After I hour. 


After 2 days, 
(at ordinary temp.) 


o.2_% sulphuric 
acid. 


Fibrin dissolved 
very h'ttle. 


Fibrin almost dis- 
solved ; Egg white 
unchanged. 


Fibrin dissolved. 
Egg white 
unchanged. 


Egg white 
dissolved. 


0.2% nitric 
acid. 


h'ibrin h'ttle 
dissolved. 


Fibrin almost dis- 
solved. Egg white 
unchanged. 


F'ibrin dissolved. 
Egg white 
unchanged. 


Egg white 
dissolved. 


0.2% nitrous 


Not 
dissolved. 


Some fibrin dis- 
solved. Egg white 
not. 




Some fibrin and 

egg white still 

unchanged. 


acid. 





A further quantity of fibrin and egg white was added and kept at 
40°C: 





After -I- hour. 


After I hour. 


After 20 hours. 


0.2% sulphuric 
acid. 


Fibrin almost dissolved. 
Egg white unchanged. 


All filn-.in dissolved. 

Egg white hardly 

unchanged. 


Fgg white dissolved 
very much. 


0.2% nitric 
acid. 


Fibrin almost dissolved. 
Egg white unchanged. 


All fibrin dissolved. 

Egg white hardly 

unchanged. 


ditto. 


0.2% nitrous 
acid. 


Much fibrin unattached. 
Egg white unchanged. 


Much fibrin and all Egg 
white unchanged. 


Egg white unchanged ; 
Some fibrin still present. 



3). In the third test, O. ig. of nitrous, nitric and sulphuric acid 
were added respectively, each bottle holding 100 c.c. of 0.\% pepsin 
solution, and immediately heated at 35°C for one hour. 10 c.c. of i)^^ 
hydrochloric acid, further eggwhite and fibrin were then added. 





After 2 hours at 40° 


After 24 hours at 40° 


0.1^0 sulphuric 
acid. 


Fibrin dissolved. 


Egg white almost dissolved. 


0.1% nitric 
acid. 


" 


" 


0.1% nitrous 
acid. 


Some fibrin unattacUcil. 


Egg white unchangeil. 



68 



K. Aso. 



O.I grm. of nitrous, nitric and sulphuric acid was again added to 
tlicse solutions respectively, and kept at 40° for one hour. An equal 
quantity of fibrin was then added. 





After I hour at 40° 


0.2% sulphuric acid. 


All fil)rin and all egg white dissolved. 


0.2% nitric acid. 


,» )) » 


0.2% nitrous acid. 


All fibrin and egg white undissolved. 



Experiment zvith Trypsm. 

Into three flasks holding 100 c.c. of a 0.5,% trypsin solution, 005 
grams of nitrous, nitric and sulphuric acid were added and kept at 40^ 
for one hour. These solutions Avere now neutralized and 10 c.c. of a 2^0 
sodium carbonate solution added and some fibrin. 





After 2 liours at 40- 


After 17 hours at 40° 


0.05% sulphuric 
acid. 


Some tihrin dissolved. 


All librin dissolved. 


0.05% nitric 
acid. 


,. 


>> >i » 


0.05% nitrous 
acid. 


Fibrin undissolved. 


Fibrin undissolved. 



lixpcriincnt 'li'it/i Emulsiii. 



This test was made' in the same way as in the last mcntionetl case. 
After 20 hours, 10 c.c. taken from each flask received o. ig. of amygilalin. 
After 15 minutes at 40'^C, the following was observed. 



studios on tlio Lability of Eiizvms. 



69 



Control. 


Odor of benzaldehyde and prussic acid. Reduction with 
Fehling's solution and witli ammonical silver solution. 


O.i^y nitric acid. 


No odor developed. No reduction took jilace with the 
above-named reagent?. 


0.1 °o nitrous acid. 


» » )> 



The result did not differ wlieii the ainyc^daliu A\as added after neutrah'sa- 
tion of the hquids. 

2). In the next experiment, the quantity of nitric and nitrous acids 
was reduced to 0.05_%', while to the control solution sulphuric acid was 
added in the same concentration. After keepin^^ for 16 hours at 18" 
these solutions were neutralized and 0.5g. amygdalin added. On keepint^ 
these mixtures now at 40° for three hours, the followintj was observed : 



0.05% sulphuric acid. 


The cliaractcristic odor of l^enzaldehyd appeared 
very plainly. 


0.05% nitric acid. 


» 


0.05% nitrous acid. 


No cdor 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 gnvips, experiments were made with 
hydrazine, mcthylhydrazine and hydroxylamine. The sokitions 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) Hydrazine. 

Pepsin: loocc o. i_^^ pepsin solution + i^^ free liydrazine. 

After two hours at 40^ ^•-% hydrochloric acid and three llocculi ol 
fibrin were added and kept again at ^,&Q. 



K. Aso. 





After -J hour. 


After 2 hours. 


After 4 hours. 


1% free 
hydrazine. 


Filirin not attacked 
at all. 


Not dissolved. 


Not attacked at all. 


rontml Ylhrin wholly 








dissolved. 







Trypsin: \% trypsin solution + \% free hydrazine. 

After two hours at 40'^C, 0.2^^ sodium carbonate and three flocculi 
of fibrin were added, keeping the mixtures in the incubator. 





After 2 hours. 


After several days. 


1% free hydrazine. 


Fibrin unattackcd. 


Not attacked. 


Control. 


Almost dissolved. 


All dissolved. 



Diastase: Sohition of o.i^ diastase + \% fi'ce hydrazine. 

After keepino- at 40°C for 2 hours some 0.1% starch paste was 
added and kept at 40X, for 2 hours. After evaporating- and removing- 
thc hydrazine with alcohol the residue was treated with water and te.stetl 
with iodine dissolved in potassium iodid. 



\% free hyilrazine. 


Blue starch reaction. 


Conlrol. 


No starch reaction. 



llmulsin : 0.\% emulsin solution + \% free h)'drazine. 

After keepiiii^- at 40X for 4 hours, o. I K'nii. amys^dalin was ailded 
to 10 c.c. 



Stiidics (HI the Lnbility of Euzyins. 



71 



1% free hydrazine. 


The odor of benzaldehyd and prussic acid 
developed very weak,* 


Control. 


The odor was very strong. 



These solutions were kept at 40X and tested in the same way again 





After 6 hours. 


After S hours. 


1% free hydrazine. 


A very weak odor appeared. 


Trace of odor. 


Control. 


Very strong odor. 


Very strong odor. 



I have observed further that also zymase is easily killed by a 1% 
solution of hydrazine. 



Kmulsin 



2) Mct]LylJiydni.':i)ic. 
I. Experiment. 

o.2% solution + o.032_^ free methyliiydrazine. 



Pepsin 

Trypsin 0.5% ,, + 

After keeping at 40*^0 for i hour was added : 
To emulsin : o.ig. of amygdalin 

„ Pepsin : 0.2^^ HCl 

,, Trypsin: 0.2% Na^.CO., 
iMiiulsin : After keeping at 40^C for 15 min. the enzym was still active 
but weakened sornewhat by methylhydrazin.e. 



* It might be olijcclod, that the diminution of the oJor might have iiccn due to the formation of 
bcnzyliden hydrazine but the control tests with the fresh mixture and that which had been testcil after 
8 hours digestion at 40° revealed a great ditVercnce in the intensity of the oilor. The formation of 
bcnzyliden hydrazine from l)cnzaldehydc and hytlrazinc in such high dilution does not take place 
instantancouslv. 



72 



K. Aso. 





Pepsin 


Trypsin. 


After 2 Iiours. 


Fibrin dissolved. 


Still some filn-iii. 


„ 4 ., 


" 


All dissolved. 



TI. Ex per uncut. 

Emulsin o.\% -f free nictliylhydraziiic 0.64% 
Pepsin o. 1% + ,, ,-, 

After 2 hours at 4o"C was added : 

To Emulsin 0.21;". amygdalin, 

To Pepsin 0.2% HCl and 3 tlocculi 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, 

(0.64^0 free methylhydrazine — P'ibrin not dissolved at all, even not 
after 2 days. 
Control : All fibrin dissolved. 



TTT. lixpcriiiicnt. 



iM-nulsin 0.1,%" + free methylhydrazine 0.(04%. 

Kept at 32''C for 20 hours, and o.i grm. of amys^dalin adtled and 
warmed. 

Immediately tested : 

Control : — Odor of benzaldehytle. 
'I'reated : — No odor at all. 



Sludk's on the Lability of Jon/yms. y^ 

IV. Experiment. 

Takadiastasc 0.\% + free methylhydrazlne 0.32^. 

After 20 hours at 24^ 10 c.c. of a 2% starch paste were added, the 
mixture kept at Afp for 2 hours, then evaporated and extracted with 
alcohol to remove the methylhydrazine. The residue was treated with 
some water. 

Control : — Alre