al

Se eaked
Sapte ae Ne

, Laie aa ee wie a ame ince ae Nt ED

Se ee ceeded tag ee ee a me ne e pengtiet 9 ane agitate Wie eae teh ee” aes >
a IE NO Ode lial tele ee a de dee tad cree tent 5 Pato Caen PE RET EE CS
Se EE ee wT SET a died a oe Ee Po Citra SiS -2 Al fth-chs
ee ete eee ee ¥ NE, edhe nee nat Sy og Sa ee See a oe i
‘ Al 5 nae ne ee Se Ee See eC SET TUTE

pag tet sys ewe ‘4 :
ak tedided ste ded or

>

ae

OP ES Pe eee eet Ae SO
Ly Pa 92 Taal is Meat meh és a aH 4 ied cee .
4 ¥. or Petr emit we Sele te aE ee eee ree et Oe OEE ae
ee ee OF a rw ne ois a — nee ee eee Pe eee ne Nealtateete ee ir Pee \ adade Fag MeTadlar =
eda dadinditady th fit dite’ san diaclet sae Sethe pe RT agp tt ag Some age pe PT It epee Oh raid

—~— 7:
oe ee ne win
vy? ate
wwe ee a te a8 " :
: FET CLES L LE Pia ee es
ROE CLEVE VTS ee ee eT wate Ke ote eie ai aan eter ee cet ene nla NE ete te eg ee wie Te Ss
Aaa ieee en” te one Seok ied eee te ete ye Ss Yr Fes pine ten te tis drone Stet: at tre ae tart
eae ee ye ee ww ee ee King Sahara tga haat)

ae died «
ee OT A

A ed rere

eee Se

irae eh deiedd eo

caneiar A Mea ig ete
ey ee eres
Sy OLE CR aS wer eer * so ee Se oe
e ve eee OE ely Saree ear ererak<?
per Tt ey er sr et - i ds tdealintaditilied se voip nena ae hee ee try teh a
eee an eeeedad ag tag he ten tt

= Seyerryr a

eer eeyr yer
“over oe = TY .
epye sr Te ¥ ee eer Fr .
st es = a a eS eel eet Seett ? ¢a
vig wir ir ee pre” err ey ree eee ne Ree ee ee
is ore? ere ‘ - > 7 Sewer eee Tee tvendesrtind? di Cee
reer Trt e : SF v7 aide ; ‘ ewer PST yey Se? ee Se Ce eRe EY
yee ey Pe Tere . j ey Severs : ond a - 3 ves , - i nee NP atte tee ee Pee ee ite eee ie ei ee eS wine ylang onetgteteeepiy ee ST ee ee hss
esse verre 4 £ ~~ re Sais eS ee ee ’ r : opie ee z Sg ee ea oy SEPT ET Te ee ee eid eet re as Se ee or 4 44> vo
vereyrre Se a ee) acta IF er wey errs Fe rrev Ff > * a a a ey eee RT Se ane aE eT ee TY frp ted eh arte ie Bs ener ve - wet wee
vere es facet Bement PO Sg See AO TE deeded dirnodet Mir . eaeey fl ORE ET SS ee ee ‘Ait ne ap ee ap cera e re sae ee ne EY ig arene wrrar co aetha he masons Og
; : period ‘ peer ety vy Sinn nite ip hg ae See eS EP yee Fe Sor ee eS ete ee SN hed ee Sane ey Tope Se ee eee or Ertan pirat rinceir4s
Lip tee eee EUS Te eT ea dirt dah amare - Re a dean deed Sere ee syste ing ar he te eke vee YET TEE MEN ET TE ffs -s-cr<b Mt fie: vndinton th > errr ae npn hae Mie
eres ee Tere eT vee ee eee LT ee ee Se ee ep yee Pee n Sow: mata tnghae nea ation inet we ama np la ee Te ETE TET TUITE ale oxy eter bs Oey rie Mite &
ied tl ii cs re Be ee rete anne miane neetere Sy te'eiy + Wet Face ere eee eee errr eer TT Seater oan
me ees SESS ETE Sean eae ip he ee ee Sse yee Pee TYE acta tity tines bss Hat CSTE tr ried iy ieie astapiarhe
a NT apd dirtd Anal tie th am dindetas ep ae Oe eee ee do trnyit Ret rteeiee Sea tse'rs
gee eT eee ET EES Y Te tr lointimigtoles

ee ceyrerEr
ete ee Se
wine nighe'e ee SONS CRS SE PET Sees

oe yee tree
eee eee 9 eee ree

er Se DE TY

ewer ed ee rerr? eve
ey Fa FETE UT TYE

ee ereyer ied

rene reet —* 3

4 rrr? ele ee ee eg Tate Oe 8 oD er virwr
cit eo OPES 1 tal te npg ~ eee ET ON ETET ENE Cee pe ee OE SOO Label
See Peete VENUE oer vt £9 eee at fe tee DROP ROLES th Sia Rice i I Sill ne ete eS
eye ere ey rey Se eee * Ae eee eee Sy SST <" a dint edly eee PETITES ae see ie edd ee eas
wr ry NN ae eee UTE owsee te ig ee EE ETE ST TY PTT e ey erry reeryTryY ee ee
pct eta ss elites Peaks ate been Fever yye vey S ee Ty vr ve ne peta he LETAATT PERG ew & er wey My Fee eT rw sere?
eee re 2 ree eet err erery. ** sever ree rv De ies ov wie ehreielelyry Sipe ve Sine dings bot de tren
. « whee he iele's = viehe Se Pee pew cles vie eT Yess

me ee ee ee
eee Yee <rer? ee
yee ee er
rerryreys

erry re ye eer HS Te

——=eyewrrrs”
Pind San OO ae tr Oe vies eer eer ee? over ®
Pat Oe estretirde eer ees eee Vey eS ee ree
fat Stel IS Bb id eee eS eee ew eT eee PEDET ETL we eee eee yg ee
sore aon weeyree®? ey ONE PE PREY TY i pia pci nies PVF =? in phe ates lh a ON OT ied
ES eel bce ew : ‘ ain Pred ae tte tO daly Sadie pr urrrr* vr eer led ww a ~ are a= wears A die
ome were rete “ ee eee sn ae arent ee ee TS ees ware Pv ee 0 ee eee eS ae retry . Oe ee ee ee TTY ere
er veetstrT? rr? gd al Sal ele al aD ES eft se of dips nS a ell F ret ee ey ee ee eer nT tn veetre erent .
siied a” = - ~ - * verre i Dy te 4 ? ee ere" ene) . 8 Hann” oe
ewieieee re re? ee v7 rer ee ae eT eE acta int ne pp tad dine efit . a vrs ete wante ee es ve le wns acitne in eMart Ta ee ee ee ee er ee ee
pg ee ee gig hd eye TET EST A ; ¥ <e Smee CSECT CTT hiatal ah iuie tetntnth tet very? a ag -rsapiing op lap ge age pet ee ae “
he . A a aes wees ; ? r* ae santapingcnat a aad é rea Ape ee ‘ - y i n't F
wearers? * ae li ha Pete ttintindrttin’ SE ee eee eT ee eer ere ene eee ey vrvey i lh ite Ae Sint iO te . s
poe reer eee TS er yy eee ee ees er tee? pata hope aS ROLES Sle he nay i Tre Dat anette dete Gr BRNO TELS TS Bled eaeaunenaneenenemn nity So ‘
errrerereT ee eteatalvia sy eure wena I rr prey Coene easyer ey ee ves Side ed ddd A TS de dc dita dented Sh ss ar dic Stn ee pli tudincvliv te de Ae ttle thd ne oe ene eee FEET ee te beh ne
ew rete re Se ad Saal ac rN a oe a Lad cial a NS Te acts Seti Ree ta Pi ete ee eS Ew ee eer ower stan‘ Pagring tg igp Tahoe nr tier ne! aan Wey ie ES C4 re ee ee ci rat tats
eT a ad rey ’ dr tir fit OES ae ne atarg re TTT ae pee 2 aa gee ee eee NN ge E eT TENET ENED TO ET a eee
eee hee re ve seyy rere Te exer Sore een ak a a a aa chen SDN a crea csistaiian talents eetaaie eee . iar diving
oo ¥ E pee werreY weet? =F thy <n de aa . . 6 ee ee ee i ead
Nee aes Seer eee Te ree ee etree “¥ vrriryr rer yt err daly det hig A Dips iy Reale tA ie Ae A a et Se
Pigalle eS eT eer ee ret SEE Ee ee eh TET TONS TO ee ee ee eer e penny werae.ve owe:
ee ewe we 'N as > = Sevres
a ~ - ewv ad hg sef ecrerrrre=ser rv _eyrrerte Tr?

ee ryvyrrerre™

ietyine we
—— *
i vere ove e'?'=
ee se acd ttle Pe OST, i dati voeins tanner tenet
; cael gt we
ge ee eT ey eee Tet osenhe et? er
- as Lot Pei 2 Rete :
one yr eels eS ee reerrerey ree. rye erry ee wig ee tee ee Ts:
SS ere ee VE ETE ey TT ern gnaw ee?) aa rate CR aad ain ET al Ahaha
wrt eet oes es? ~~ wer ~ Z
eT er eeeT tt ewe 2 tes er ore a Cee ie wee ve va ee yew eee is eicivte 2 0te
i.e > SP cts ok a cect AF cin ta Sain nan PSY ET Th Si ca eure ee Rie Sir ae Gath x =e
‘ TY owe? re ew : :
: ee Baglin et Barns Xe we eeverTrTeTt eee ee ee eee Fee
oar’ oe eT ata rt ee we Pere ree = Tha ieee daca pas nla pen hac Be
oirtelenielae tise eS SS oe TS earere yer? eT Bae Ragin Bh sae Bins sire Se PSS eats te ;
Oe Eee eR EET TEER . 2 tir rx talin te rah Saas Ho ST a dindirte Deiat ae ee ee eS re ele me:
peer ere Te Cv ET ee petites OST Th Fe PRET ECLET TE eee cane eet eycner ee pes pureipentet nae
ict tn intr eS ie Ie Lactate eee De ane were? een eea ee er ere ee rer ek ee Ve ee es ree tga det) ve "rrr
cae id ainda 6g <r ties hie? . re STEVES STEVE TUT Te arewrr nS a, he tb ie Stee Leet a lobe sen
ee ee Eee TSE Pee ee eee eer? F NT lon eer? Ute eS Fee T e ee eee ET ERLE EV VETUT sad atin: 54 00 Sit
Pe a OT dort lint te Ee emaree ty A im finn AS Ch tend Minin Al Senne Singin SSA TE Be pi haat sad ay tied hal Pa kdig A RS “ y « ree
ww ee eee ye aed EEE eee nT by ete Tenet See a «le — 4 1a in ote dlncpain He - aon
were tit <ae we rs . -e sen © eh eevee ee ee tee ee as : .
lia ee a : & Ayer tested ae : . . m <r wert
: ve Se teteh tore wee fig Bh Sy ticat Sa e e Dadictincle Aad Sted oe 7: — Sarre 4 Siow "
verre eee Pere eee eer re oe “e : wartime As yl Tara Ay dinette Betlaehae Smee ¥ en
a ee Mae ach thd a Pa a . ? ’ eteeteewverrTry Tyee SS fo a ek ee --— ee? sees > >
eeyeyvEetsyy? et ee ee eee ee RE CELI ERT ES OE ee we 4 “ oh te de r , es ie rome
te cnt Cot nae «1S tet ‘ eite aa meee . a sree Gare
a apa. we . ; ay
sew Y : : Caae poe ak , :
eee er eRe es tT . pk: beatin ete
I ai -
A —<+- iy i over
= ee renTese Sooo oes . =
Conve so “ > an ( , dept - °
eeugree? Latins ah ee datade tated So
eeverrrrt : s :
eetyrerre™ - : a —
weoververresrs” ee : r “: "a
ee ererrt eet ee : :
ae ée - - - - ft
92 oFete on . 7 “
eevee rt wees =
_—~eevererT
pr doe Sp Oe dna : ;
oe "3? e7? “ -
essere™t —"*- - <"er*? .
ona ee ererrererrr? NT oe oe © . ; *
Ain thin pany te allay “ owerverr en = ms
y= ‘ 77
hin eahling =.
=—se 066 OFe « ever = a)
ee ~ .

Historic, archived document

Do not.assume content reflects current
scientific knowledge, policies, or practices.

UNITED STATES DEPARTMENT OF AGRICULTURE
BULLETIN No. 405

Joint Contribution from the Bureau of Plant Indusiry, WM. A. TAYLOR, Chief,
and the Bureau of Animal Indusiry, A. D. MELVIN, Chief

ne,

e Washington, D. C. PROFESSIONAL PAPER December 5, 1916

| LUPINES AS POISONOUS PLANTS

By

_C. D. MARSH and A. B. CLAWSON, Physiologists, Drug-Plant
and Poisonous-Plant Investigations, Bureau of Plant Industry,
and HADLEIGH MARSH, Veterinary Inspector,
: Bureau of Animal Industry

CONTENTS

4 Page Page
| Parti—Introduction . . . . . . 1 | Part I1f.—Discussion and General Con-

; clusignstis Ses oe . 28
Part I1.—Experimental Work . ... . 4;\' Diterature Cited) 3:02.05 Pe a 43

WASHINGTON
GOVERNMENT PRINTING OFFICE
1916

=—Iae*

wh ge tam
a ae aKs

UNITED STATES DEPARTMENT OF AGRICULTURE

BULLETIN No. 405

Joint contribution from the Bureau of Plant Industry
WM. A. TAYLOR, Chief, and the
Bureau of Animal Industry, A. D. MELVIN, Chief.

Washington, D. C. PROFESSIONAL PAPER December 5, 1916

LUPINES AS POISONOUS PLANTS.

By C. D. Marsu and A. B. Cuawson, Physiologists, Drug-Plant and Poisonous-Plant
Investigations, Bureau of Plant Industry, and HapieiacH Marsu, Veterinary In-
spector, Bureau of Animal Industry.

CONTENTS.
Page. Pages
Part 1.—INTRODUCTION.....---------------- 1 | Part III.—Discussion, EtTc.—Continued.
Summary of knowledge of lupines as Toxicity of different species of lupine.... 30
POISOHOUS Plantse =e 3-2 ---2------ 52 1 Toxicity of lupine leaves for sheep.-..--- 30
Disimipmtion of lupines ..-/-- =... ---- << 4 Toxicity of lupine seed for sheep...-.--.-- 31
Common names oflupines....--...------ 4 Toxicity of lupine pods for sheep-......-.. 32
Alkaloids oflupines...-.......-.--------- b) Toxicity of lupine fruit for sheep.....--- 32
HeimOkene nas soles tee seer e cS Sic Sa 6 SYA pPlLOMSe LM. s sais Si aatese ae coe ee 33
Part I1.— EXPERIMENTAL WORK....--------- iG Pathologiyerecc ccc coas este eae Se seas 36
Pharmacological investigations by Soll- Comparison of ‘‘lupinosis”’ and poisoning
TOROS ee AS 2s a eee 7 of sheep by American lupines. ......-. 36
Field experiments with lupines......-... 13 Remedies sieves saiclteees eam eenye 38
Laboratory experiments with extracts Range conditions under which sheep are
| oflupine seed upon mice. ........- eae 25 POISONCO et ee eens asap e sees tases 39
Part IIIl.—DiscussioN AND GENERAL CON- Treatment of range animals to avoid
CUURTORG A823 Saas Sas Se ee ences 28 POISON Rete eee ae cetera i eee ere 41
Failure to poison sheep in 1910, 1911, and SUMMA age ss naan nee shee cess leat 42
HONMS SAE SIRS Se So An SS Se eee ee eee ON}: eed BaP PALURS (CHING bois SORE Sp eedeShooSaonBaeanae 43
Lupine not a cumulative poison....-..-.- 29

PART I.—INTRODUCTION.
SUMMARY OF KNOWLEDGE OF LUPINES AS POISONOUS PLANTS.

, EARLY HISTORY AND LATER STUDIES.

The lupines have been known from very ancient times, and are
mentioned by many authors, e. g., Theophrastus, Marcus Portius Cato
Censorius, and Pliny. The last-named author (ed. 1856, pp. 49-50,
452-453)! treats of the lupine at length, especially with reference to

its use as a green manure. Several species have been used as culti-
vated crops in Europe, more especially for the reclamation of sandy
soils. It has been used also as a fodder crop, and the seeds ground

NotE.—This paper will be of special interest to the stockmen of the West.”
1 Bibliographic citations in parentheses refer to ‘‘ Literature cited,” p. 43.

52191°—Bull. 405—16——1

2 BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.

into meal have been used both for domestic animals and as food for
man. In Germany the land utilized for lupine, according to the
latest available statistics, is 346,753.3 hectares; on 200,000 hectares
of this amount it is cultivated as a green manure. The poorer people
among the Greeks and Romans and the Cynic philosophers made
use of lupine meal in bread. The bitter principle was recognized not
only as disagreeable, but as injurious, and the seed was especially
prepared in order to get rid of this property. Among the Greeks the
seeds were cooked until soft, to remove the outer skin, then placed in
sacks in shallow places on the seashore to wash out the bitter prin-
ciple. Afterwards the seeds were dried, ground in a hand mill, and
baked into a poor bread. Only the poorest people used this meal
unmixed, but others mixed it with other kinds of meal, making a
more digestible flour. (Landerer, 1852.) Because the lupines were
planted in Maina and there used for food, the people in that region
were known as “Lupinophagi.’’ Lupine meal was also used by the
ancient Egyptians, and is still used in Andalusia, Corsica, and Pied-
mont. (Cornevin, 1893, p.314.) In modern times lupine meal, after
a process of ‘‘Entbitterung,’’ has been used to some extent as food
for animals. |

As a medicine, lupine seeds have been used since ancient times.
Pliny (ed. 1856, p. 452-453) enumerates 35 different uses. The
main uses, however, seem to have been as a cathartic and as a vermi-
fuge. For the latter it was used as an external application as well
as internally.

Bellini (1876) reports in detail cases of poisoning in man from
using a decoction of lupine as an enema. He states that Averrhoes
and Hofman pointed out the poisonous properties of the plants, and
that Paullus, 1708, reports a case of poisoning of a boy by an enema.
The reference to Averrhoes and Hofman could not be verified, as
apparently they only mentioned the plants as a vermifuge. The
symptoms mentioned by Bellini are dyspnea, defective sight, dilated
pupils, and stupor. These symptoms, as will be seen later, compare
fairly well with those of poisoning by the lupine alkaloids.

Isolated cases of poisoning by lupines were noted as early as 1860,
but it was in 1872 and the following years that heavy losses of sheep
occurred in northern Germany. While there is evidence that some
animals are poisoned by the alkaloids, most of the cases, and prac-
tically all of the losses, have been from the use of lupine hay and are
caused, as will be seen later, by ictrogen. The occasional poisoning
of cattle and horses reported in Europe appears to have been from
the use of the seed and is alkaloidal poisoning. Sheep are also
poisoned in this way, but the great losses which have stimulated the
extensive investigation of the subject have been by ictrogenic poison-
ing of sheep,

Qa ee ee eee ee _

LUPINES AS POISONOUS PLANTS. 3

In America the first published report of losses of sheep is by

Chesnut (1899, p. 404-405), and this report with the papers of

Wilcox (1899), Chesnut and Wilcox (1901, p. 100-110), and Slade

| (1903), comprise all that has been published up to the present time.

Other authors have mentioned the subject, but their material is

| all taken from the reports of Wilcox, and Chesnut and Wilcox.

The first general account of lupine poisoning in America is by

| Wilcox (1899). A much more extended account is given by
Chesnut and Wilcox (1901, p. 100-110), with details of cases and
symptoms. Wilcox notes that mature plants are the more poison-
ous, and Chesnut and Wilcox definitely state that the toxic principle

is mainly in the pods and seeds and that lupine hay should be cut
after the seeds are shed. Both Wilcox, and Chesnut and Wilcox

| apparently consider the poisoning of sheep in America as similar to

the disease called ‘‘lupinosis” in Germany, although Chesnut and
Wilcox (1901, p. 109) state that the ‘chronic form of the disease”’
has not been recognized in America. Sollmann, in an unpublished
report of a laboratory study of American lupines, a report made
under the direction of the Bureau of Plant Industry, which will be
discussed more fully later, states that he failed to find evidence of
the presence of ictrogen and gives details of the symptoms produced
by the alkaloidal substances extracted by him. His work was not
connected with field investigations, but the results, viewed in the
light of present knowledge, clearly indicate the probability that the
field cases of poisoning by lupine in America are not cases of
“lupinosis”’ in the sense in which the term is used in Europe, but are
the result of alkaloidal poisoning. The field investigations reported
im this paper establish without a doubt the fact that, so far as ob-
served, all cases of poisoning of range sheep by lupines must be con-
sidered as due to the alkaloids and not to ictrogen. These field
investigations are in entire harmony with the preliminary laboratory

| study by Dr. Sollmann, although carried on in an independent way

without reference to the preceding results obtained in the laboratory.
ANIMALS POISONED BY LUPINES.

While the losses of domestic animals have been mainly of sheep,
other animals also are poisoned—horses, cattle, goats, swine, and

_ fallow deer—and laboratory experiments on small animals indicate

that none are immune to the effects of the toxic substances.
~ Wilcox (1899) states that in Montana a few horses have been

| poisoned, and Chesnut and Wilcox (1901 p. 100-110) give specific

instances. Poisoning of horses on the range or in pastures is not
common, but is known to occur. Dammann (1902) states that
cattle are affected by the alkaloid, and cases of the poisoning of
cattle on the range have been reported in America, although the
cases are not very well authenticated.

4 BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.
LOSSES FROM LUPINES.

The losses from ‘‘lupmosis” in Europe have in some years been ©
very great. In 1880, in Pomerania the loss of sheep was 5.89 per
cent (Cornevin, 1893, p. 316). It is stated that in some herds in
northern Germany the loss was as great as one-half to three-fourths ©
of the total number. |

The known losses in America are very heavy. Chesnut and
Wilcox (1901, p. 106) tell of one case, among others, in which out’
of 2,000 sheep trailed over a region covered with a large quantity of
lupine 1,000 were sick and 700 died. In another case 1,150 died out
of 2,500 (p. 104). Losses of several hundred are not at all uncommon,
and occasionally the deaths may be nearly 50 per cent of the whole
number. More complete knowledge of the losses has been obtained
in Montana than in any other State, because Montana has been under
close observation for many years, and it is very possible that this
State suffers more than others, but more or less similar conditions
are found in other western States in which the mountain regions are
used for sheep grazing. It is probable that the lupines rank as a
close second to Zygadenus in causing losses of sheep. :

DISTRIBUTION OF LUPINES.

While lupines are found in the eastern United States rather spar-
ingly, and are there represented by only three species, in the Western
States there is a large number of species, and these frequently grow
in large masses, so that sometimes they are cut for hay. They form
an important factor in the number of forage plants. Horses seem
to be especially fond of them and will sometimes eat the green plants
with great greediness.

Of the species of lupines in America, only a few have been used
in this investigation. One of these, Lupinus sericeus, is illustrated in
figure 1. A thorough systematic study of the genus is in progress
by the Bureau of Plant Industry, and further work is being prose-
cuted for future publication, dealing with the characteristics of the
different species from the standpoint of chemistry and pharmacology,
as well as from the standpoint of field experimentation.

COMMON NAMES OF LUPINES.

The lupines are known under a large number of common names. —
Among these are sundial, old-maid’s bonnets, Quaker’s bonnets,
Indian beans, wild beans, blue pea, and blue bean. In some sections
the name loco is used, but this is generally by those who do not
know the true locoes. 4

LUPINES AS POISONOUS PLANTS. 5

ALKALOIDS OF LUPINES.

_ No investigations of the alkaloids of American lupines have been
published, but the European lupines have been the subject of ex-

Fic. 1.—Lupine (Lupinus sericeus).

tended study, and the literature, especially of the chemistry of the
alkaloids, is very large.

Cxcola (1834) attempted to isolate the alkaloid of Lupinus albus,
but probably did not obtain the pure alkaloid. Landerer (1852)

6 BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.

obtained a substance that showed a beginning of crystallization,
which he called lupin. Beyer (1871), found an alkaloid in lupine
from a study of Lupinus luteus. Baumert issued a series of papers
from 1881 to 1889. Baumert (1886) summarizes the knowledge to
date and announces definitely the presence of two alkaloids in Iwpinus
luteus, lupinin (C,,H,,N,O,), and lupinidin (C,H,;N). Schmidt (1897),
Davis (1897), Callsen (1899), Schmidt (1904), and Bergh (1904)
summarize and bring the subject to date with extensive additions
to the detailed chemistry of the alkaloids. Schmidt (1897) states
the alkaloids as lupinin (C,,H,,N,O,), lupinin (C,H,,N), ‘ dextro-
rotatory-lupanin” (C,,;H,,N,O), and inactive lupanin (C,,H,,N,O,).
These are found in the lupines examined, as follows:

Lupinus: aneustitoltus:. 22.2.4 5.5 ee dextrorotatory-lupanin.
Tapinus.albusse 2 ct ae ce ee { dextrorotatory-lupanin.
aa inactive lupanin.

upmus luteust 2." aes ee ote se

lupinidin.
Lupinus niger... te. a 2 ee

lupinidin.
Lupinus perenitss seg ose ee dextrorotatory-lupanin.

Lupinidin is identical with spartein C,;H,,N, (Muenk, 1914, p. 394).

Callsen (1899) gives an extended account of the chemistry of the
alkaloids of Lupinus angustifolius and L. perennis. Willstitter and
Fourneau (1902) give experimental evidence that the formula of
lupinin is C,,H,,ON. Schmidt (1904) and Bergh (1904) in extended
papers discuss further the alkaloids and state that another alkaloid
is found in Lupinus perennis, oxylupanin (C,;H,,N,O,). }

Liebscher (1880), Lowenthal (1888), and Raimondi (1891) give
details of the pharmacology of the lupine alkaloids. The action of
all is practically the same but differs quantitatively. Liebscher
states that lupinidin is 10 times as active as lupinin. There is a
direct paralysis of the medulla and cord, the respiratory center bemg
first paralyzed, and then the vasomotor. This is accompanied by
weakening of the heart. There is a fall in blood pressure, a slowing
of the pulse, and dilation of the pupils. Death is produced by
asphyxia, with which are associated convulsions.

There is no curare action. The dose required to produce any

symptoms is ordinarily fatal. The alkaloids are less poisonous for
mammals than for cold-blooded animals. Small repeated doses pro-
duce no effect, nor do they establish a condition of tolerance.

ICTROGEN.

Early in the investigation of the poisoning of domestic animals in
northern Europe by lupines, causing the disease which came to be
known as ‘‘lupinosis,” it was observed that these were not cases of
poisoning by the recognized alkaloids of the lupines. Lupinosis has

LUPINES AS POISONOUS PLANTS. al

a definite line of symptoms distinctly different from those produced by
the alkaloids, especially characteristic being a hepatitis, which pro-
duces a jaundice exhibited in the conjunctiva and visible mucous
membranes of the lving animal. Kitihn (1880), Roloff, (1883),
Arnold and.Lemke (1881), as well as others, found that the intoxica-
tion known as lupinosis could not be produced by alcoholic extracts,
but was produced by the marc of the seeds; if the poisoning were
alkaloidal, the reverse would be the case. Dammann (1902, p. 348)
states that the plant often becomes more toxic on keeping. More-
over, while the alkaloids are always present the ingredient producing
lupinosis is inconstant. The plants raised on some fields always
produce poisoning, while on others they are harmless (Raimondi,
1891). It was found (Dammann, 1902, p. 342) that the substance
producing the disease is insoluble in alcohol, ether, glycerine, and fatty
oils, and is soluble with difficulty in water. It is not readily destroyed
by dry heat, but steam under pressure makes it harmless. This
hypothetical substance was called ictrogen by Ktthn (1880) and
lupinotoxin by Arnold and Schneidemthl (1883). It has only been
recognized by its physiological action.

Ictrogen is not considered to be a product of the metabolism of the
lupine, but to be the result of the growth of microorganisms upon the
plants. This explanation is not. based, however, upon any experi-
mental evidence, but is reached by a process of elimination of other
possible theories. This is the theory advanced by Dammann (1902,
_p. 341-343). Other theories of the cause of lupinosis have been
advanced. For example, Ziirn (1879) propounded a theory that the
_ disease is produced by microorganisms in or on the lupine leaves; in
other words, that lupines do not cause the disease, but simply serve
asacarrier. ‘This theory has not been taken very seriously by others,
while the theory that the disease is produced by ictrogen and that this
substance is produced through the action of some unknown micro-
organisms upon the lupines is quite generally accepted as the most
probable explanation.

PART II.—EXPERIMENTAL WORK.

PHARMACOLOGICAL INVESTIGATION BY SOLLMANN.
SCOPE OF THE WORK.

A series of experiments upon Montana lupines, identified as
Lupinus sericeus, L. leucophyllus, and L. cyaneus, was conducted by
Dr. Torald Sollmann, under the general direction of V. K. Chesnut,
then in charge of the work on poisonous plants in the United States
Department of Agriculture, and following is a report of the results.

Kighteen animals, rabbits and guinea pigs, were fed upon pods
and seeds with no results, none of them eating enough to produce
toxic effects.

8 BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.

Extractions were made of the seeds and pods, and impure alkaloids”
were obtained. The experience in this work leads to the suggestion
of the following method of extraction:

Moisten the powdered drug with alcohol containing 1 per cent of HCl; pack in
percolator; macerate with alcohol; percolate with alcohol until the percolate gives —
only small turbidity with Mayer’sreagent. Mix the percolate with sand and evaporate
the alcohol. Treat the residue repeatedly with warm water, until it gives off only a
slight reaction with Mayer’s reagent. To the united filtrate add Mayer’s reagent to
complete precipitation. Wash the precipitate, suspend it in a little water, and decom-
pose with H,§; filter.

Evaporate the filtrate to a small volume, add an excess of Ca(OH),, filter, exhaust
the precipitate and filtrate separately with ether, as quickly as possible, neglecting
the last traces. Evaporate the ether. Treat the residue with absolute alcohol acidu-
lated with H,SO,. Let stand 24 hours. The precipitate will contain the lupinidin,
the filtrate the other lupine alkaloids. These will require further purification.

CHARACTERS OF THE ALKALOIDS.

Physical characters.—The physical effects of the aklaloids make it
likely that they are similar to those occurring in the European
species. Several of these characters were observed in the course of
the isolation of the alkaloids.

The (impure) alkaloids were obtained as brown oily liquids of a
strongly alkaline reaction and a strongly bitter taste. They were
easily soluble in water and in alkalies. They were slightly soluble
in petroleum ether. No crystals were obtained. During their sepa-

ration a strong odor resembling conin, characteristic of lupinidin, ~

was perceptible, especially when strong soda was added. The
isolated alkaloids were almost odorless, but again developed the odor
very strongly when 10 per cent soda was added.

Chemical characters.—Strongly heated, they boiled and evolved
dense, white fumes. Strong sulphuric acid, cold or heated, alone or
with formaldehyde or bichromate, gave only browning. Strong —
nitric acid also gave no characteristic reactions. |

Precipitation reactions—The alcoholic solution gave partial pre-
cipitation with sulphuric acid. The dilute neutral aqueous solutions
of the chlorid or sulphate behaved as follows:

NaOH (1 per cent): Slight precipitate.

NaOH (strong): Precipitate, partly soluble n excess of the soda.

(NH,)OH: No precipitate.

‘Na,CO,: No precipitate.

Mercuric chlorid: Amorphous precipitate, easily soluble in excess of HCl. The
precipitate was incomplete, giving further precipitate with Mayer’s reagent, and also ~
the lupinidin test with alcoholic sulphuric acid.

Picric acid: Fair amorphous precipitate.

Tannin: Precipitate, soluble in excess.

Iodin in KI: Good amorphous precipitate.

Pot. ferrocyanid: No precipitate.

Pot. bichromate: No precipitate.

LUPINES AS POISONOUS PLANTS. 9

Nature of the alkaloids.—The alkaloids were not obtained sufficiently
pure to make definite characterizations possible. Their close agree-
ment with those obtained from the European species in physiological
action and fatal dose make it very probable that they are closely
related, if not identical. Lupinidin was fairly well identified by the
conin odor, by the insolubility of the double chlorid formed with mer-
eury, and by the insolubility of the acid sulphate in absolute alcohol.
The precipitation by these reagents was not complete, so that there
must be other alkaloids present, presumably lupinin and lupanin.
This portion of the work needs further elaboration.

The yield of crude alkaloids in the extraction was as follows:

Specimen IV: Lupinus sericeus, parasitized pods, 0.02 per cent from alkaline ex-
tract; 0.133 per cent from watery extract.
Specimen V: Lupinus cyaneus, seed, 2.462 per cent.

Gerhard found from 0.5 to 1.2 per cent in the European lupines.
TOXICITY OF THE EXTRACTS.

The various extracts, prepared as described, were administered to
rabbits and guinea pigs, by mouth, stomach tube, and hypodermi-
cally. The symptoms were practically identical, and will be described
later. It was found:

(a) That the toxic principles must be alkaloidal rather than ictrogenic.

(6) That the fatal dose of the drug (as extracts) to rabbits by stomach, in the case of
the seeds of Lupinus sericeus and L. leucophyllus, was between 30 and 50 grams per
kilogram, with the seeds of L. cyaneus between 70 and 100 grams per kilogram, and
with empty pods of L. sericeus and L. cyaneus over 100 grams per kilogram.

(c) That the fatal dose of the crude alkaloids (in the purest form in which they were
used, from Specimen V) lies, for rabbits, gastric administration, between 1.2 and 2.4

grams per kilogram; for rabbits, hypodermic administration, between 0.123 and 0.246
grams per kilogram (agrees with Léwenthal’s (1888) figures for lupinidin and lupanin,
viz, 0.2 and 0.4); for guinea pigs, hypodermic administration, between 0.052 and 0.1
gram per kilogram.

(d) That the alkaloids are five to ten times as toxic for rabbits when given hypo-
dermically as when given by the stomach tube.

(ec) That guinea pigs are about twice as susceptible to the poison, when given hypo-
dermically, as rabbits are.

(f) That repeated administration of the poison to animals did not increase their
susceptibility, as is shown by the fact that rather prolonged feeding was not fatal,
_ and that no tolerance is produced in this manner, as is shown by rabbits 77 and 78 A.
The animals in either case, after having been injected repeatedly, died from the last
dose, although these doses were not very greatly above the fatal limit.

SYMPTOMS OF LUPINE POISONING.

Tt will be useful to describe the typical course, which occurs with
only minor variations when any of the extracts are administered.
The symptoms set in with a general depression. The animal is
very quiet, sits flat with ears laid back; the respiration is rapid,
labored, and irregular. The temperature is not altered in a constant
manner. After a time it is noted that the animal, while apathetic

azi9t-— Bull, 205—16——2

10 BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.

-

if left alone, is quite excitable when disturbed. The reflexes are
heightened. The movements are brusque and exaggerated. The
depression gradually deepens; the animal partly loses control of its
hind legs, so that these tend to drag. Often the animal lies on its
belly, the head on the floor and legs spread out. It is still able to
walk. Itsoon loses this ability and sits or lies still. When disturbed,
it reacts with shivering, and becomes more and more convulsive.
Stimulation now produces more or less violent spasms, the first spasm
being strongly tetanic. The animal may assume the strychnin posi-
tion, with legs stretched out, and back arched in; or it may rise on
its feet, the back arched upward. The spasm soon becomes clonic
and incoordinated, the animal pawing the air in an aimless manner.
This is followed by relaxation, and the animal remains quiet unless
disturbed, which disturbance would cause another spasm. After a
time the convulsions occur spontaneously. In milder cases there
may be no general convulsions, but twitching of isolated muscles—
of the face, ears, neck, back, and extremities. The head may swing
in a rhythmical pendulum movement. The respiration during this
convulsive paralytic condition is slow, shallow, and irregular... The
pupils are variable, but need not be dilated (which speaks against
the view that they are asphyxial; the mucous membranes are also a
bright pink). Many animals urinate copiously. The animals may
remain on the abdomen or recover a sitting posture for some time
after the onset of the convulsions, but after a time they fall on the
side. The respiration becomes gasping. Asphyxial convulsions set
n, and the respiration stops from half a minute to 1 minute before
the heart. 3

The depressant and convulsive symptoms agree with those de-
scribed by Lowenthal (1888), Gemma (1882), and Raimondi (1891)
for European lupine alkaloids. |

It is not possible to make any general statement as to the time
required for the development of the different symptoms, since this
is extremely variable. It is remarkable, however, that a consider-
able time may elapse before any pronounced symptoms appear,
and this even when the solutions are administered under the skin.
It is to be noted that doses which are little below fatal produce only
very sight symptoms.

Recovery may occur from any stage and is usually so complete
that the animal gains on its original weight. The intoxication leaves
no post-mortem lesions, gross or microscopical.

OPERATIVE EXPERIMENTS WITH LUPINE ALKALOIDS.

The effects on blood pressure, respiration, etc., were studied on
five dogs, anesthetized with morphin and ether, and arranged for
tracings. The alkaloidal extract of Specimen V (seeds of Lupinus

LUPINES AS POISONOUS ‘PLANTS. 11

cyaneus) was injected into the femoral vein, in progressively increas-

ing doses. ‘The dose is calculated as grams of drug (not of alkaloid)

per kilogram of body weight. Arranged by doses, the effects were
briefly as follows:

SMALL DOSES OF 0.1 TO 0.4 GRAM PER KG.

Respiration: First quicker and deeper; then somewhat slowed; irregular, and shallow.
Carotid blood pressure: Rise, fall, rise, normal.
Heart: Rate somewhat quickened; strength somewhat diminished.

TOXIC DOSES OF 0.5 TO 0.75 GRAM PER KG.

Respiration: First quickened and deeper; then somewhat slowed, irregular, and
shallow; may stop.

| Carotid blood pressure: Rise, then great fall; effect of sciatic stimulation lessened.

Heart: Rate first slowed, then quickened; when vagi were cut, slowed; strength,

weakened.
VERY LARGE DOSES OF 0.9 TO 6.25 GRAMS PER KG.

Respiration: For a few moments deeper; then very shallow and stops.
Carotid blood pressure: First slight rise, then great fall.

Heart: Rate first slowed, then quickened, then stopped; strength weakened.
Convulsions may occur from 1 gram up.

DISCUSSION OF EFFECTS.

Respration.—The respiration shows a short stimulation (increase
of rate and depth), followed by depression (slowed, irregular, shal-
low); with the larger doses it stops before the heart. The action is
probably on the centers, for it occurs after section of the vagi, and
when stoppage has occurred it can not be revived by asphyxia, slap-
ping, or stimulation of the sciatic, or injection of saline. No recovery
occurs from even a just fatal dose after an hour of artificial respiration.
The respiratory center is the first vital center to give out completely.

Blood pressure.-—This shows a short, moderate rise, followed by a
more lasting fall, which is quite marked with the larger doses, even
those which are not fatal. Although the changes often coincide with
respiratory changes, the two are not interdependent, for they may
occur independently, and blood pressure changes occur even during
artificial respiration and are not influenced by the latter. Whether the
changes were central or peripheral was not investigated directly; but,
from the fact that when vasomotor paralysis exists stimulation of the
sciatic is sometimes effective and sometimes ineffective when asphyxia
is effective, it is rendered very probable that the action is central.
The vasomotor paralysis may precede, coincide, or follow that of
respiration. It may be partial or so nearly complete that the pressure
sinks to some 20 millimeters with a good heart action.

The heart rate.—With toxic doses there is first slowing, then quick-
ening; with minimal and maximal doses, there is usualky quickening
only. The slowing and secondary quickening occur equally well
when the vagi are divided; they are therefore at least partly periph-
eral. They are not always accompanied by changes in the strength

12 BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE. 7

a)

of the heart, hence they are probably not muscular. Further, when
the heart is quickened by large doses, electric stimulation of the
vagus has but little effect. The drug may therefore be said first to
stimulate and then depress the vagus end mechanism. It may a
a similar action on the vagus center.

Strength of the heart. Ts is affected only by rather large doses,
_but it is then always depressed. Experiment 87 shows that the life
may be kept up with a complete vasomotor paralysis, if the heart
beats well. This tends to show that the stoppage of the heart, which
forms the final cause of death, must be due to paralysis of its muscles.
This is confirmed by the fact that it can not be revived by injection
of normal saline. |

Convulsions.—Convulsions were observed in but two of the five
cases. This may be due to the anesthetic or to the difficulty of
securing proper dosage. When observed, however, they preceded
respiratory distress and were not connected with any change in
blood pressure, so that it seems that they are produced by a direct’
central action. .

Skeletal muscle—The existence of convulsions in the last stages, as
well as the result of stimulation of peripheral and of the sciatic, chow
that muscle and nerve are not paralyzed. |

Cause of death—The followmg vital functions are eventually :
paralyzed by the lupine: Respiratory center; vasomotor center;
cardiac muscle; the last named being the last to become paralyzed ;
and being the faa cause of death. t

SUMMARY.

The results of the investigations are in many respects incomplete *
and emphasize the need of a more exhaustive chemical and pharma- —
cological study of the American lupines. They have shown, how- —
ever, that these lupines contain alkaloids which are toxic or fatal ifa
sufficient quantity of the plant is consumed, but which are harmless 7
if the consumption is below a certain limit. Up to this point the
lupines may be a useful food if due precautions are observed that the ;
limits are not surpassed. These alkaloids could also be largely ©
removed by leaching with water. The ictrogen, which is especially —
feared in Europe, was not found in any of the American samples. It ©
would be of great-importance to determine whether this ictrogen is —
uniformly absent, for if it is not, much more stringent precautions —

would be necessary. :
CONCLUSIONS. ,

1. Feeding with the lupines does not produce any symptoms in ~
rabbits and guinea pigs, as a sufficient quantity is not taken in this
manner. }

2. The injection of extracts, by stomach or skin, is fatal if suffi-
ciently large doses are used. :

ar

age

LUPINES AS POISONOUS PLANTS. 13

3. No ictrogen was found in any of the six specimens.

4. The toxic constituents are alkaloidal and seem to agree with
those of the European species.

5. These alkaloids produce a stimulation and then a paralysis of the

| following structures: The respiratory and vasomotor centers, some

convulsive centers, the vagus end mechanism, and perhaps the
vagus center. Large doses given intravenously paralyze the heart
muscle. The convulsions do not appear to be purely asphyxial.

6. Pronounced symptoms are seen only when almost fatal doses
are given; smaller amounts do not produce any conspicuous effects.
When death does not occur acutely, there are no late effects. Re-

_ peated administration has no influence on the action.

7. The cause of death is paralysis of respiration. Death occurs,
with hypodermic administration, in 12 minutes to 24 hours; when

_ given by stomach, in 10 minutes to 3} hours. The symptoms set in

only shortly before death.

8. The fatal doses for rabbits by the stomach, figured for the
original drugs, are as follows: For the seed of Lupinus sericeus and
L. leucophyllus, between 30 and 50 grams per kg.; for the seed of L.
cyaneus, between 70 and 100 grams per kg.; for the hulls of L. cya-
neus, and L. sericeus, over 100 grams per ke.

9. The fatal dose of the crude alkaloid for rabbits, gastric admin-
istration, lies between 1.2 and 2.4 grams per keg.; for rabbits, hypo-
dermic administration, between 0.123 and 0.246 gram per kg.; for
guinea pigs, hypodermic administration, between 0.062 and 0.1 gram
per ke.; for dogs, intravenous administration, about 0.012 gram
per ke.

10. The fatal dose for rabbits is between 5 and 10 times as large
when the alkaloids are given by the stomach as when given intrave-
nously. Guinea pigs are more susceptible to the alkaloids than

_ rabbits when the solutions are administered hypodermically.

11. In the treatment of the poisoning, artificial respiration was

found useless. Good results were obtained with potassium perman-

ganate, diuretin, and tea.
FIELD EXPERIMENTS WITH LUPINES.

Field experiments with domestic animals have been carried on for
six years—in 1909, 1910, and 1911 at Mount Carbon, Colo., and in
1912, 1913, and 1914 at Greycliff, Mont. Two species, Lupinus
comatus and L. myrianthus, were used at the Mount Carbon station.
At Greycliff most of the work was done with two local species, L.

leucopsis and L. argenteus, but two feedings were made with L.

leucophyllus collected in Idaho in the Caribou National Forest. Not

only were different species used, but feedings were made of different

parts of the plants and also with the plants at different stages of

- growth.

BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.

14

“HOLOTISB,)
Od
“dure,)
oO
“dureo

pues YaVq sqqoM

*S ABI)

od

‘0d
"HOPOTISBO

‘0d
“UMP [CEL

‘0d

‘OT

"OJOTISBE

‘Od

; og
“a Ure;

“poureyqo SVM

posn quryd Yor
Wor) UOTL BOOT

‘Pyotiaay 10 FL6L pun ‘eleor ‘Cl6r Ue pun “oj0,/) "UOgun ) JUNO, 10 TL6T pun ‘OL6L ‘606T We sniidnT yn spusunrsedxa Burpoof fo hunmungy— | aTAv yy,

£

000'T 49.

JO 1USTOM

8 "FOL
861
611

o SE

88S

L'99

“spunod

OOL LT

06

PTL

T LTP
FOS

12

i PES
8gT

© 6LT

SOT
by

L'8E
CLL
GPP
*spunod

*peuTUe

poyeys
oy posn
spunod

ae ee (OO ee

ee ee ODE

> K1DA090Y
= "YyRod
~ ALOAODO YY,

“YINSOY

trees sQprttt:
Serre topes:

ops

Se) SOUOINT
‘opeydyns
ULNTSOUSdRI,

eee se OuON
‘oyeydyns
ULNLLSOUSRI,

“Apoulo yy]

wees 8 sop:

sees Qpe:

“sess 9p:
steep:

‘op:
"* YOIS JON

BEET iia}:

>" MOIS JON

sees sQpt:

eons 11S
“"YyRoqd
aa SOLS

“ssouyyt
JO AYLOAG

81 ,, AUp,, OLOYM Sosvo UT Jdooxe “VYSIOM Wedd OF Poonpod Orv OSOP JO SouNsY YY Yq ‘[eMoyvUL Aup UOATS 010M

aoyoy OP]
--"qinay pure ‘sureys ‘SoAvoryT |”

07 ON OTL

roprctfrcs ope

Bg? Soe eh i ee eS ODee sat CMA non na Our

Shoes sevens SUOMOT PUG ‘SUIOJS ‘SoABO'T |°" ">" "EZ OF Gg OUNL

see ene Cee ee i ie ae a ASO} aru see uO py COR
‘O16

ak aaa ie po qinay pure ‘suroys ‘soavory | ~*~ ~"6Z 04.9% “Wdog
“6061

siege rn ee) 0 ale large “= gy 07.9 Ayng

Rennie yinay pues ‘sromoyy ‘sur9ys ‘soawory |°° ~~ > ° 9% 07 ST ounL

te TY alle gear es ha spvoy poog |°**"""9% 04 21 ‘Sny

hl le RE MMI SIRE GIT SR GR I CCE Coy oy OVO OS OE TE Co} fo rosin yt

Sco ae Se SIOMOY PUR ‘suId}s ‘SoARO'T |" T “SNy OF 2% Ane
“TIGL

miaie win ps sagehs:eliein/s)=Lalgiereiis ade. ~"e sind "opr" *|"77" ZT 04 FT “4deg

sible)» wie toleip.e wicieie qinay pure ‘suroys ‘soavory |° >" "" ET 07 OT “4dog
‘OL6L

“-""9% pue gz ‘ydog

ASSN IN Se aE Ba NTS AR Ea ODe yan == 9g 01 270 1d09

pac? Ser \ynay pus ‘suomoy ‘suroys ‘SsoaworyT |7 "EZ puw Zz “ydog
“606T

*(poyeqs OSIM “yuOur :

-1oY}0 sseyun poy) posn yueyd Jo par g

“110d x0 JO 07BCT

66%
“CLI

>

G "PES
8gT

G Ger

Ts

€&

GB “8S

bg

9°06
“SpUunog

*posn
yueyd jo
VUSTOM,

SOT 94 SIT
G8 0} 98
0} 08

O&P

OOL
OOL

000 ‘T
000‘T

OOL

009

OOL

009
*spunod

“VUSTOM

{pojou

‘UO

+ eeths = SBTT "ON
Sis
wr aglte “TOT ‘ON

ate

‘ON

pet aks Ss FI ‘ON

SSmyDUuLod
“7 Wears dovyg—*(T

le “029 ‘ON

cre eeeeeee Gn *O

isnyjuniihu

sietaintatete mint eco 8) ‘ON

J8NJDULOD
“T GOALS SOSIOFT—

a Abie P21 ON
Se. FELON

pee: "8 ‘ON

‘ON

‘ON
‘ONT
‘ON

82

UOT} BUSISO(T

*[euUTIy

JOQUINE [VIS OY} Surpoooid (4) Ystaojse Uv Aq poyYeUsSIsep speuraTy

JSnyupiihue
“T UWOATS SOSLOF[—"y

15

0d POL" pects gots Mee ODase Alcs MOIS ONT [orcas see "(Weiq UT) punosd ‘poog |-""7"77 777 6T “Bn . Beg |erereee ==" 06, :
‘od coe imiehose ee op’ ** gsr eOHON Sie OPses EVsas estas (urei3 Ul) punoigun ‘peeg opened eeoe ee a0 ee Scaunensecaal on
| ‘(youeip) |
0d CPSeee es Awede| ec yyeeq | prow oruuey, |-""~" UJCOUL Ga eos ee ee cr ap se apa gee aS aie ee ets) ea LGSes 2b 2 Stig "10% °
: eer oe CG PL SON AC UNE ip re LN TL 10Z *0
con oe ; 2 len one eee eum |e EIOIS AION” | earieh ateiee (uviq Ur) punoIs “paeg |-""7"7- See TODS = 5; 02° PE HOWTO S| SA ee ole ON
u 19 Ig (Uyeis UL) punoisun ‘pesg --*"* op eeiCT SV ial SE Ry: ZI; OREO) = ot | 2 aera 602 “ON
“S161
aw 38° So ee Es Es ee paeia onkconiey) (merase Tees) eee 1G ae g0: 6g Lissa “O8T ‘ON
ne 3 i ieee re | aerial Peele lh aa Oe re ee Sere: POD sees "Z “ydag 07) 7% “3nV Gee le Wey Cais Fey) eos ZLI ‘ON
i On aac eee eee eee erga See ae So ena PEDROS Sates See Celi iene COLO Ay, OT ye | 0800. Chases ances ZOL ‘ON
a aa Beye Oe ale zece=--ceee| os een ee A eee Me PRS ee aloe Oh ee op See ry oa "PI “sny | CFO % SOS 2 Ne een 891 ‘ON
Ky ou Ae eee et eee ee gels eee oP ue eee Eh eh (YoueIp) FOI] ieee RCs Ve Cea 26 iis beet eo [elena es CLI ‘ON
7, oe Bee Sas iven eral a. ae joie seed Be Pao pet an ie gia SUI0}S PUB SOABT |" ~~" 7" €107g8'sny | G's ZG)309 $09" 1a paces €91 ‘ON
5 oe Olen? ees eae rete eRe dla wae ies eee ete (doo W100 UL) AIO | See Ops: tan eke eae) ee eL1 ON
: ed Le et ee BBE As a BU Neal oie c tsa ie Atm SU ea Cenc a ee et cL “ON
| i SSE PA riponinse 9 Sy | ate OD GS nie ODE rel ede g emer aie eee ee Mele UNBORN ae aes morn eRODAIGA | éI NE he g | ee 9L1 “ON
5 SO ald ie re ae oe eee BO ole Aree ate eS ae eee OD GE 10h (03°81 alee ae ILI “ON
AOS MR LRG ne <2 i ete a aca, 0 AOD eee ROD GO| a rt cl ae thr nite ret mretenn ee WODinaa seal cork te rape ODiees: VET Th Seen aetng ere ere OST ‘ON
iP ee nee Aleeip gh -nctd |ersiem oN Bese els CREE ao aee pOienenintie caves cei ewe OE TREG ‘sny 0706 Ate |G 'T NRedeetere ever cc LL ‘ON
2 on AT es vests oe tel eee CEs als cas Dene aaie titiae he Mela ot aes Opis “"gydegozge Ame | $28 | #18 92 %06 [oo POT ON
FOTiGis Pestle OTS aa ke epee aes een Sires Weaetiecnia rec gee ; 7op "7772607 9T Aine | OL CD SOUR Ala aise 191 ‘ON
2 l BI $661 P op {INI PUB “SUIO4S "SOABO'T |-" "~*~ 46041 Apne | G2 °ST CO 2OVE CLs elas naa ents OLT “ON
a “CI6T Jsasdoona}
: Sve eres aC eeiaal RES ee Berar Se Rous eee ea Alte teak Glad on NST eae Nonna ‘T UeAIs desysS— H
ey BESPI SOG o8 ee ass op ek le SOOPE Pert ee Re eee eee OHONGT 2tiy |e 109, || ONL OUNCG ha ovI ‘ON
a . eranicat egy ee aaa BP 4 ABGO'T €¢ 01 G “ony Gor GEE CAS ALG s| viet aioe 9FT ‘ON
< Pp es can ay Seed ae eel nee a On Regd iN ae a Le “FI “3ny 0708 ATNE | Gz °88 VANDA an ZVI “ON
HOG gOnOra gales iice trols seg D Eel Lo, CODES ob Rin carina etch we ee Reg aUDp Shiela nem Ops Dg ER SCT a dlr ee 0
ee od sae | eee Seve Reon ODE 1255 aan Op Faas “qinay pure ‘sIeMOV ‘suIe4s ‘SaAavorT |= => 8% 03 6g Ang cI or age ce oN
. od F901 op OD ns aac oes SIaMOY PUB ‘SUIO}S ‘SeABET |--- ~*~ 61 078 Ajne OOS | eipGarst ate Role Ser OFT ‘ON
7 cen
eee ere ed ete era enna Clee te Wire oe Oe coke eure he ‘T UaAts deeyg—'y
MS ee BCE al seaters dl en ODA Sule PU sales sha aetna tle ate op Hew ee HOUOT SO vial. OU OY 4| OPT Oly Cha ares ae LEI ‘ON
3] 2 (OS pra Sucre oe cate SIE Suni Disc Sule sea Die Gee ie ee "7 SpBey pees |" "6 “sny 0762 Aine | —¢ 09 OGIO # IZiial|= area EFT ON
od G80 Tal inden Gece eee ORAS Ain dies Op' |) ) 135117 37 t amyy pace “sureys “seAwory [oo " 5 re Opss > 92 Ree ean fees are ecT “ON
on owe Be aria le cce ae vel eee rises eden re ee eine eee Ace oleae re Ceres Oe Ss SS Fae se sceae ey OFT (ON
. Gos | areata bos.V ie pes Nee cee OG SEC Ae Sil Wade eect Poy NR Tien: See one Oh |G ey ee ISL, CORE AINE Nee (1H es Skee OS) gi | Peep eee tere 3 ‘O
od 8 SL CURES | euikaee OD es eee Op7**|777 77 741May pure ‘siaMmoy “Sur94s “Seavey |""9 AINE OF Rg ouNL | or eo Sou ae ee I ON
we He reer cetenscs ORs alata ODT alanis aoe Sonar, “ae gene Mae Eee EO Diy Sali ud eats Ona KG ORT re lage eae 6E1T ‘ON
ee op op SI9MOT PUBS SULOJS “SOABOT | ~- >>” cf 04 ZT euNnsg Iv WE rea eer ak (ates eet oF SEI “ON
- “TI6T = ne
; . See aynr Weer oe fee Weare Rake oe feral =f alee io eye Caan eta See ene me “TT UWOAT 90 S— WL
od We be elie omer a pect ODE ilicenee WOR ee eel eee Se eer ge) we Utes 2 01039 °4d9g | ¢°6T Uf OAL Ley RPE oe 6II ‘ON
od (a eps larger acetic QOS ant SUURS ei Wambo st : "---qrmag |72 -ydeg 076g “3ny | gag OD OM (Wot pee COT ‘ON
oa TOMES | oe on tel a ESOP ET Esha PBODEAS Case SIOMOG PUB FMI |**** FZOLVEL SNY | G's 88) 01566) dl catcemenaes &ZI “ON
a 18 op op ; SIOMOY PUR ‘SUIO}S ‘SeAVE'T |“ TT “SNy 01 0g ANE 18 (Nay LL COE oer e ass” 9TT “ON
SSNYQUDILMUL

‘T WoAIs dooys— A

pHing

rea]
on
Pp
isl
=
=)
Oo
=
=
i)
<f
Gy
i)

NT

Ss. DEPARTME

.aes

5)

BULLETIN 40

16

“Od SPS 'T
‘od P9L'T
“Od GCE 'T
‘od Thr *
‘od G88
‘0d Tbh
‘0d G88 *
‘0d B99"
“UOT }BIS | 188"
‘0d 088°
OUR] “S010 01
[BUONVN NoqIeg | OPP’
d ‘Od OPP *
‘Od (ccm
‘0d ae
‘0d 900°
‘Od T98 °
‘0d OSPF”
‘Od CVS"
‘0d 98e°
‘0d Thy"
‘0d. GOV"
‘0d 6VG
0g cTg"
‘Od Obb”
‘Od LEE *
“UOTYBIS JIPOACTY | 61Z°0
*spunod
OOF 49d
; *[CUITUB
*poureyqo sem JO {USLOM
posn yuvrd yoru poyeys
WOly UOTIBOO'TT | AOJ posn
spunog

Y
Pere AO Done tee Sia COLO NBN) | enemas O Dau linia eae ge ag Pee Re ee a ODE Aie game eee Fe 0% Ayne
“urdoaye
yqgvod | puw yesopyg |" aywod |r ic gee tae se Pa ie he OPO cor el ec “6 Ane
SEAR Beene PEaipeangn sce uslecieres SO per le uenne ent ue beb cmon einsie malaise nO pcan lense eon yn |
a ae en sileee ET IO Dyanna |b OLSHLO!NT dl tars necarss cae stay mae cee noe acl nse SA CIe es) TD cre ee (petra cesar Baer TRACT BUC
*(possaidoep)
Magan Tee essiseeeessoprrr] SOLS JON |*(suLpooy pooszoy) podoypoaop ATHY “yrnay foo gy Ate
roar eee DD Bey eo oy oS ae Oa era ole 9 al fe ai IERIE Tu Ga yocs yy oy) {sy OVE U IT GYONOY fal. | [pen aebs! 2h OO RLIEG} oy 9O
Ser tacgire teeta [eRe ERO De wee oh tee fae 8 AO DF ae l| Fes Sa he RGR eee ae ICO) 0), Mania | erg wel “OF ouns
Tree ae a Sea ILLS 0} oY peti || aah Pi(0) 0) aaa i rare SCG OT ““"(qoueIp) SOABOTT [7777 tOpr ts
Peasitabs Toeeeeyesomessoprr cy topes" """"""(qoueIp) sunod ‘spvoy JemopT |--"""""""""""g oung
“PIOL
Pope ene yPenier SOME. ce sors ropr tty sstttscrrsrss"=*(ueiq UL) punois “peog |"---6T puw gt 4ydog
a at ah 2 ig ee Ar Teen Oo} 0 nee ACT GUSH gy 0 a SSPE ee rani Eas Dee eet SPROUL DOOM comes tae ks ct ‘ydog
PR CAC. Beal Goi eel AOR taro ake ene ae IC caters tet enor netaeuy tetera eit ten Sam CD TD eathe ti (AC ee TERT TO) Teed CLO
Pees pista | peace ON AEN Fae OP teal nen pipe Sraattien a gone ca eR OD ee sarees ais Seek 1 ydeg
te athe | Sanat EMO PO bares pa ard aD ees [ieee ran recs each ie asses ae aa Sierra) Trae ia en Tea N CLO
Ap Nacsa eae y| trate wees eho Roce at Oprah ts CE tai hs an a aa ag H pia 0} Oa ote GUS ahah 2, yous aces
Ouse a te Vara O Dee FT OLOTS COONTH Reena Cc Cis Sin mite etre niaiaed sald phot, [Opera lieyce'h 5 gaa “eT ydog
Tere AO Ogee Rae te cao) TE eal PHAC0) oY aia full eal tire Ae Oe there Re SIRO PD NMUS Eee Seer ONO
SPA TOMNODON TI: |pmiitie Ol Pine tal |erererp ois >FHYIR [PRS LOe Bates Od a RE ODse ease CPUS “z ydog
TORT OR Sais a SON) aeneielie a SLOLSa ONE es tienes Pees RO TOeag Se O Dear eae lev creo SS ESO peak
mea MOD. Clas | past <eteran OTD por gras nell aad yyeod, jor Fhe aati hime OR Tp LCR EE SR pe: NAO) DIS Ios cards Sete ebay
Seat cee Ee OMe Fe aOR palemn racine selma ae sor OD Maan rn VOCeDILG Ogera nis
BOE pc opel eae eaearag 0) 0) an TE oO Rieter Se eed ae **(ueiq ur) punols (B908 |7"" "FS PUB Es “SV
MALES aa pea | Sheth Of peter | pats erg eo) 0) LOO Sr Tas “= (upers UL) punodssUN ‘poog j7-- 777 > fa © [0 jane
Bop rope aids aa] (avai bg'(0)/ 0) areata eaina oc 0} 0 Leg can ea ORI ARE Sa Pe) Diy a OP Hae scion eee COU
Ri SESE tale EOD ES kn piso ieasss/ OT Saige |ererara Se SE Se SARS get eve tM 0 OF st sae tara Siti Pat ODie ese

“NOY

*Apoulo xy

-" QUON |" YOIS JON

*ssoul]r
Jo AjLtoaeg

‘

*(po}e1S OSTM

-19Y}0 ssoyun poy) posn yueyd jo yaeg

‘ponurju0)— Muoyy
‘Puypohosy 1 FL6L pu ‘ST6r ‘C6 We puw “ojog ‘uoquny qunopy 10 TIGL pun ‘OL6L ‘606T we snuidnT yun spuauasaduxa burpaaf fo hanurng— | atav J,

“yOu
-T10d x9 JO 07RC]

>
1821 8 ASAP RS CNT
GLP BOGE oe, SRR Rsk scoiirias 8lZ ON
O61 'T ZRS SOR 060 ipikeme nem LLZ°ON
ele" AOS OU pee ole tase “163 “ON
OLN 6L 0} 88 77°77" "26 “ON
Chr Gi \(0) emer 2S “""* 082 ON
698" RSG) ©.) > ae ae SS LLE ON
ggg: (OSE OWN Tk walter a 0G (ON
PRE" sy PAOYE COFNLS) thea pes Seo “69% ‘ON
ssisdoonay
> "T udATS dooygs— M
8£9° 431 Fea “*Z06 ON
Les* OG) OURO) ae ena "S61 ‘ON
le ae
‘T Goats dooys—
oge* SHOR AIm cmsetar ens O12 ‘ON
CPS" OF: CRATES |Faans "°°" Q6T ‘ON
98" GO ONL Baia | eeisiaes “10%, (ONT
P00 * 99) Of 07k |i ast Sen GreaG
“8N0JUIDLD
‘7 UoAts dooyg—'T
Le OL OUMROU | Sceve casas 961 ‘ON
G8P* COTKOT NRO Te | trees “€0% "ON
G0Z Oxh: OFF FOR). [paves sar “21% ‘ON
ZS" BQO) sO 2ii Raia """"Z0Z ‘ON
Z8S* PO) sg, Pe ee C8T ‘ON
618" SOL ON) G7) a | Rae rlevickas 961 “ON
FIZ" OUP Oley OG) | Penaiinsiae 11Z “ON
£9" ORMOTNYp Ru. F Peace nes 802 “ON
LIS LO OR CL itsse mess “061 ‘ON
POL” Gar OlesR()Tealt eens “" 20% ‘ON
190 Te OURS or |r “*° 861 ‘ON
2 SPUN0T “SPUN0T
“posn “VUSTO AA UOl}CUSTSOCT
queid Jo
VUSTO AA - >
peuruy

17

COX G fi TG eae ie meaty pentane Te een ae Meta s O OP ease is
a es eee i OPS aise SR ES eae Seg sae OD ee Se ee 3
ie eee ae ape po9o010 fe) Gurpee} . Ae ae -
eee coe eee oe J) Peey POUR DeM.u4 sped edlNe| ee. “eSny | ove: ee.
ees ae Ce ee apr ee ses ee eS eS
aa eee eee gyocccnchcnenenece eee ops ee ee:
Mee EE Te ee ey eee eye ee cee Drittefecectesses[sesestecessesefeceeseeees seca.
dn et ee ee op Bogiat Else oa koe cis a oe ee ee ee :
eee ieee Be eee C0) Oty bee ee ae Be eee Shoes| ee eee ee ei ON
ee ee ee ee ob cede Seen mae | ie eS ee ae ee Geet or
Ge ere op Aiea ert eae ere, Menno en cnar en eee ee |
eg (ocioucoocp eee ers |or see gees aeeas SRP ee [o)j Oneida i ciety ede erie SO Seeraa ten eee ss ane £8e “ON
: . eae a Opie ccececceecceeeseseenseseent es OPsgseale= sess ta of setae coe Cee lecmea te ee 2 “ON
ees Dea) Se ec eee 0) Spee eee vane Op 1 a ite ON
Eee eee ane SONS anh ee eee Bee ae OD Ey tac Se Uae an ee ee 06 “ON
" ee oahu nee Opec ccc eccceeeeeeeneeetoes ODES sees sae oe a een
ee eee a ODE seas 2 eee ob coc ea ee I) ovens Ree Ose “ON
ee tee oe a Gan ee eats ODEs adlepes es we a gee Lie “ON
gg Sonccufioccicyocs oe Sel ere re ee CO) OE Gar nol He rece a Sas ae Sie ee tea ener ene ae zee ze “ON
: ee ees ae eter ee ODE eines ee Op ee ee eee 122 ON
: eee es ae ache oe Ge oniet ween ee epee OD ems Sloe ce op eee i “ON
g ee Wee Ghee. | ea one a ew OD Smeal eee a a eee sic ON
: a ee eee eee OBE EUs | Pie aa pated nreae, pape Dae ieee cere te ee ee
: eee eee gi eet |isuec aceeseots rscegtenl eterna OD ince ee op Se ie ON
Z ag ococdhcciaee pee Ghss:[ie-adeatescuarees cee oem OPgees [ss ers Be ee ee ioc “ON
i iam icc eee ie op mssecgpes-|-scercesesseeseceseeeeeereess er O Deen | a epee eg ee
: abe ob Siseigpers|prec it inesctseeeeceneeesst 9p Se ye aes eines ape son ee ee ee 86 ON
ia eee eee oo op op Se ee ae ae Sees OD apse eee et ee Bess leerats cf ee cee Loe oe “ON
; i ue ees cee eae 0 Oe ee ee ce (©) ORS 2a Eee oe Sr a8 se Be See ee ee ee ese “ON
Se eee ee ere a
nee spo fac ghey cc Sg eee oe
Seer - seefeosecestecteececietstectictecstiopesces|esrectsesseecgpersesfeeserscned[sereeeseeerees[eesescescs z-ON
f od Seesaseen ase sacar Oke 32 doe <= IRIS ON, | Sates 5 i ees ee ees yp oe ee orcenes eb | yes
z Oi eee ee ope : Vv nlkeees Nee ee eee eens op sg Se es ee ox
Se eS ee ee a
: ag Eos ee pe ka Pe eee eee OD RSS 3 eS eee an Be she is eee eae 196 °0N ‘|
: “g ees Svebeg [eon ee ot ames ae (Oyo Ante Ra aCe Soe He Sous tsb aces ak Ove “ON
so Pecererae a se Cyc eee epee ee tae enacongeut poe i einen anes Rn Gaeie ss 186 “ON
- aa ener rape nee ae ODevea lsc a eee a ee eS at "ON 7!
oo law ee fag Yo} euldny] WO pepe, OATS Gita b tte reat Se Ge on ‘i
es eee =. Op***| -posy peoloj) PeAoTe. poses ae 55 s Oe a wx :
Be ore tee ae |
‘od e911 wseee (youep) YOISJON | peoroy) pedoferop ATTN} oon Deel aals ae | so es Yee a :
“od copy fttttt yea | prow opmury, |----qyeaq, |7+777 7° ee : von
oe ae Sea UG es See ee Se ee . ;
Le ee ISJON [ccccccetetecceeeeeeeeneec eee Fs pe ee ee ce ee a
ce i Op” *| 262 T 768 04 406 v2 16E “ON |
ane teu FEO) Des Se Ree Sei re Ale : 3 6 ise E
| roa 0¢e T COE ONEE Ria ia ear aera OFZ °0 a
od €&h 1 acy epi ee |
POG OLUUe Te: SOIS AONE |S sees ene eee rE Ret AS :
wet eeresoperses|ereeseseee ee
. ez Aine | €88'T
G6 04 Pe ep 69Z “ON

BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.

18

‘od

HOTIRIG

*pouteyqo sea
posu yuerd Yor
ULOLJ UOTPROOT

ThO°%
Tg¢°
Thr”
S6r°
V0G"G
LVL 'S
LV9 'T
166°
908 T
Teg
€6E 'T

122°
POG 'G

T99°
GGL SZ

“spunod
OOT 49d

“BUTTE
Jo VYSTOM
poyeqs
IO} posn
spunog

ae Meeaalle sateen “+> =-0ps=-y-(uypo9y pone) padoqoaep A1ny AMAA BOCROOUR TSH LO: AHKo ts)
soso
BESET aa
9yeuog.aeo
piel ae ives S BTCC CILTID OR i= TOLSMON( = ent) nee snus "7" (SUTpoo] pooloy) poog |-*--- >"> * “i OD eatiae
(uoT) sod ;
“Tpur ef
-T SSO)
ree Ss ewe alse e p= OTR eS "| 0 SOTSMONT || Pees ane es Rae enn rier OO MEA Too o> Ses Oar
CISION ok rebe es aon eae “Op> >>|" (Surpooy art eee AN Nae NH NAuEEE ||PSOQRR OSS “1g “dnVy
*(Soso
poywodor
o}eUuo0gaeo
Pca Cakes -1q wmntpog "se scopr ttt rset sss 2*"(SUTpeey peoroy) poog eet nso} of eee
a ah Ee En ein “--"QUON |* “01S JON |7(Surpooy poo10j) podopoaop AT[NJ “nay | 6s BY
“UlOJ
-jeo pur
oyeydy ns *(SUTPOOJ PodJ0})
a ee op***| uwmrsouseyy |"-"*~"Op**"} poos Jo ysour poys pey yey} spod ody j--*-* 7-7 Opes"
Soa niceXae [lee ae ODe sais ARO CINE sore RO re Segre ou sak (0) olpmecdy elena tg gr 8% “SNYy
CARRERE Aa Benes (Oy Olena SIRES oo Pa aOe OS OSE OC OL (joni oless toy n (Opi) ol\e}sfllo mee ieseae ag lay
EES mee athe scans CO) oVig aie ears SU 0} 0) ira ae ecani tn pi ig Rote (WBIq UT) Peed |" "°- "7" Ops”
pape Oy carats a eae (Oy Ose aaa at po] ON al | aesccrekm aiectaerley arian ane aR CRSA OF(0) ONT at ietaaeelaees aya dems ee Lf
gaptescinisiotoore No} lca ges= (OY Oe OBIS) Ah CET og fp ISERIES FOE SIGN RTL Mo) ONE rte IGS teria oA tet
sae cal eso a OD ees oe ano Dems emmccinn ers(S eT OOT POOLOP ISA GO) |na nes 9 ae Ose NY,
SCOT OES Qe O TSO OINTs | ee aac oases sicaeiaik hs (ogee) tout) jokes}: ese RS es OOS Sao o Sees
acies TOC Cs ee eRO Di ae eee at Gada ot samme Oak (OUTO GOT DOOLOM INGA Ons: ten os aces Os een
* AIQAODO3T |-° >" *” OD ares BHO TS ait isie ss cian oustgiesnie ics --*(SUTPOoJ PodlOJ) Poog >= ST ‘sny
PRR oO gins (Wh OPE | HENS "Cal oN Aid a re EE ceed har fee ace yas pec pes OO) gy tk eee, “ony
*(surpooy
Poiana de NC ons op-****|"777*"0p***} pooros) poos poys pey yey} spod ediy |--*°-**- "TL “sny
Nr aS eae QDs saree? son | ne mene cos s(STILN OG PanLON SeAGoOT) (pect ae aera ON OLY,
*(suTpooy
Pe roe ay akan ae oUON |""" ">" "Op"**| poddlOJ) poos poys pey, 7eyy sroeaie Pessiaeisie (rere rea Tv
oul
----"ygvod | Aystyas ‘pore |" -- 77 Yeo | -poey pooloy) POAOUIOL Poos YIM spog |--77-7 7777 7 g -sny
*Ssouy|r *(Ppe7VIS OSTM “quour

“4INSORT,

*Apoulo xy,

Jo AYLIOANG

-10Y}0 SsoyUN Poy) posn yuvyd Jo qaeg

‘penuryu0)——" ruopy
‘Pyolias) 1 $L6L PUY ‘EL6L ‘CI6L We pun “opog “uoqung qunoyy 1 TI6L puv ‘OL6T ‘606T Ue snudn'yT yum syuaeursadxa burpaaf fo hinmung— | wav,

110d X09 JO OFBCT

L69°T

66h *

OOP “Tt
96h “T

SOP
966 “T

$99 ‘T
LGP*
EGE *
£¢0°
BELG
G6E
Loe
690°
890 'T
866 °
G80 'T

Spot
860°
Ted °

GGL ‘T
"SPUNOT

*posn
que[d jo
JUSIOM

Sy ON PRe [8° ° = === OReONin
§o8 02 4%06 [°°"°"°""° "$9 ‘ON
Ps Ol AGh) + hexoses 19% "ON
OR O1'RCG) Rats =e “""G8% ‘ON
8109 BR. eee Saas 162 "ON
OS Osa eee "" 18S "ON x
18 (Arp) 108 “ON
OO harry hud 2 Eo ae 892 "ON
4O)-" Ol GOR. asl Geese ieee 1S “ON
BAG ON AC hl eras P16 ‘ON
MTG -Oh-26) sees 06% "ON
Ole ae, Siete 99% “ON
Ub, ON OL. oh ree anaes 16% “ON
OG: ON: see J Sepia ra se p22 “ON
TR eae ry. | PE as GhZ “ON
OG> a5 ae eee incites 1&% “ON
#94 0} 28 "(AIp) ES “ON
18 0} $8 (AIP) 08% “ON
QaON Ca Pens Name C66 “ON
Feor 0 GOT |*** (AIP) Lb% “ON
HCO setae cla ena £90 “ON
“SpUuNnvg

“VUSTO AA “MOTYVUSTSO(T

‘yeuurUy

\

19

“ONNUT @ OVW 1

LUPINES AS POISONOUS PLANTS.

0d (Seis Paes Seal lage ere |S es Ope ees oes OP Se ee ee eee ears ODF sear ean 01 -3d99g | 6F0'F 468) 104. ~ 26s sal os woe oe 108 ‘ON
0d TA SARE PGS aceite cee Opseiwe|s SOTS AON ewes | eens (SUIpoey Ped1OJ) SPABET | -------- “26 °SNY | 1hZ"e O6 O98 Gia ewes aces £93 "ON
*(SUIpsej Ped10}) pees
0d OGG oe alo eee eG Es OD aera gsmmojdurAg] pus sIEMOL Moy @ pues ‘sue}S ‘SoAvETT | -- Go “SNYV | 619% COIL OH JANE ies ees eez °0 N
“snaquabio
‘T wears deoys—"]T
ONG) [sates RS Se Ra oe a ee ae 20) Op Sa FESS ae ODS=s| So esas elon oe SUI0}S PUB SOABVAT |------ OT ‘4deg (i) Be eae | eee S ee Cho (ON
0d OLAV eee eee C0} Opteaise [seared 0) ol ee an ie oe aie ee eS Ops cS eo ee 6 “4deg | 999°E ya, OM AS ch erie 908 "ON
0d Ste Oe eerie ee eee ouON 2 STOISMONT pera aun gy yeas (SuTpee} peodoy) Seavey | ----- L“ydeg | 66°F SOLES es | ae aes 066 “ON
urdo148
pure (sesop
peyeeder) |
9} 80 q Ivo .
‘od SLL “oye | -1q ais po SSO Cla nee. ea eee a a Oe eee eee ae g 4deg | e¢.° i Sieraenne ee pees 69% ‘ON
*(Saso
prieadan)
97eu0q Ivo
‘oq ic eeeiee eee eee Iq wintpog |--*"~" Opty 35 a SET (SUTpee} peotoj) peeg | ----- op---*! 62° The, 08 208: 5. eee 966 ‘ON
= =

ak

20 BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.

Inasmuch as nearly all the recorded losses are of sheep, most of
the work was done with these animals, there being, all told, 141
experimental cases. Two head of cattle were fed, and there were
10 experiments with horses. Table 1 contains a summarized state.
ment of the field experimental work.

EXPERIMENTS WITH CATTLE.

Two head of cattle, Nos. 108 and 620, were fed with lupine, but in
neither case was there any result. This should not be interpreted,
however, as proving that cattle can not be poisoned by lupine;
accounts have reached the Department of Agriculture of the poisoning
of cattle in the field under circumstances which make it seem very
probable that lupine was the cause. However, it does not seem
probable that the lupines cause any serious loss of cattle.

EXPERIMENTS WITH HORSES.

There were 10 experiments of feeding lupine to horses, 8 with Lu-
pinus myrianthus and 2 with L. comatus. Horse No. 72, between
September 22 and 26, 1909, ate, per 1,000 pounds of its weight, 77.2 ©
pounds of lupine, including leaves, stems, flowers, and fruit. The
animal died, the marked symptoms being general depression and
dullness, abdominal pain, twitching of the surface muscles, and high
stepping as he walked about. The animal was very much consti- —
pated and in the autopsy there was clear evidence of intestinal im-

paction. At the time, because of the negative results obtained, it was
thought probable that the intestinal impaction was the cause of

death, but a review of the case, after more complete knowledge of
nine poisoning, made it appear quite certain that this horse was a
victim of lupine intoxication.

The two feeding experiments with horse No. 124 were negative.
With horse No. 78 there were five feeding experiments with Lupinus
myrianthus—two in 1909, two in 1910, and one in 1911. Im 1911
there were also two experiments with this horse of feeding L.
comatus. In four of the experiments with horse No. 78 there were
distinct symptoms of illness, consisting of contractions of the surface
muscles, intestinal disturbance, indicated m the main by constipa-
tion but showing diarrhea in one case, dullness and depression, with a
tendency to lift the fore feet unusually high when stepping about.

The experiments with horses were sufficient to establish the fact
that these animals may be poisoned by the lupines. No deductions,
however, can be drawn as to the toxic dose, since the periods during
which the feedings were continued were extremely variable. To estab-
lish definitely the dosage for horses would require a large number of
carefully conducted experiments comparable with those on sheep, and
for such experiments there has been as yet no opportunity.

LUPINES AS POISONOUS PLANTS. adh
EXPERIMENTS WITH SHEEP.

In the experimental work with sheep the attempt was made to
have the animals, if possible, eat the plant. In experiments with
seeds it was found that the extremely bitter taste led the sheep to
avoid them, but when they were ground up and mixed with bran
the material was eaten quite readily. When it was found difficult, in
the case of corral animals, to get them to eat enough to produce
any effect, drenching and forced feeding were used. In drenching,
the sheep was placed on its haunches and the drench, which con-
sisted of the plant material finely ground and suspended in water, was
given by a bottle. For the forced feeding the material was ground
and given by means of a vetermarian’s balling gun. The plant mate-
rial fed in this manner was placed a little at a time in the back part of
the sheep’s mouth. As soon as this was swallowed more was given,
and the process continued until the desired quantity had been given

to the animal.
TYPICAL CASE OF SHEEP NO. 184.

Sheep No. 184, a 2-year-old ewe weighing 62.5 pounds, was taken
into the corrals on August 16, 1913, for feeding. On August 17, at
8.45 p. m., she was given 200 grams of unground seed of Lupinus
leucopsis, mixed with oats, corn, and wheat, which was equivalent to
0.705 pound per 100 pounds of animal. During the day this was all
eaten.

On August 18, at 6.55 a. m., the sheep was found lying partly on
one side with her head on the ground, as though in sleep, and trem-
‘ bling in the forelegs. When stimulated she attempted to get up,
but was unable to raise her body from the ground. Through the
struggle the respiration became more marked and heavy, the animal
breathing much as though in a deep sleep. After the struggle the
pulse was 174 and fairly strong. At 7.10 a.m. the respiration was 36
and the temperature 101° F. Plate I, figure 1, shows the condition
of the animal as she appeared at 7.15 a.m. At 8 a.m. she lay with
her head extended on the ground, in much the same position as at
7.15. She raised her head for a few moments, then slowly dropped it
with eyes half closed, asifin sleep. At 8.05 a.m. she held her head
up, and apparently made an effort to walk. Her respiration was 36
per minute, noisy, and deep. Her sides trembled as she breathed.
The pulse was 122. At 8.50 a.m. she got upon her feet and stood sev-
eral minutes, then leaned against the fence and sank down again. At
8.55 a. m. she stood with her legs bent at the knees as though in an
effort to balance herself. Her respiration was 36 and of the same
character as before. There was some frothing at the mouth. Her
ears hung low. Plate I, figure 2, shows the attitude of the animal at
8.57. At 10.30 a. m. she lay breathing as before and trembling.
Her temperature was 103.3° F. At 10.35 a. m. she was raised upon

22, BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.

her feet and then tried to walk, but was unable to stand, and lay down
again. At 10.47 a.m. she was unable to get her breath and went into
a spasm, in which her legs straightened out very much as though she
had been poisoned by strychnin. At 10.484 a. m. she was breathing
again more naturally. She lay upon her side, trembling violently,
with her hind legs extended rigidly. At 10.50 a. m. she was quiet
again. At this time she appeared to be a little bloated. At 10.52
a. m. the muscles were rigidly contracted again, and the trembling
became more violent; this was followed by a spasm, in which she had
great difficulty in getting her breath. At 10.55 a. m. she quieted
down again, and her respiration became more regular and deep. At
10.56 a. m. another spasm came on. At 10.58 a. m. she was quiet
again, with pulse 174, strong, and regular. At11a.m. another spasm
came on, but the animal soon became quiet. At 11.08 a. m. the head
was thrown back, the heart fluttered, respiration stopped, and at
11.10 a. m. the animal was dead. For several minutes before death
the heart action was weak, and just before death she was unable to
get her breath. The muscles gradually relaxed, the head was drawn
back, and the heart stopped.

An autopsy was made upon the animal immediately, but very little — |

abnormal was found. The heart stopped in systole, and the brain
and spinal cord were congested. Other than this the animal appeared

to be normal.
TYPICAL CASE OF SHEEP NO. 201.

Sheep No. 201, a ewe weighing 71 pounds, was brought into the
corrals for feeding on August 16, 1913. On August 17, at 9 a. m., she

was given 100 grams of ripe dry lupine seeds of Lupinus leucopsis, the —
seed having been ground in a coffee mill and mixed with 100 grams ~
of bran. At 9.30 a. m. this material had been eaten, and she was —

given 100 grams more of the ground lupine seed, mixed with bran as
before. At 12.20 p. m. most of this material had been eaten. Her

respiration was loud and deep, as in a sleeping animal. It was esti- ©

mated that the animal had eaten altogether 175 grams of lupine seed,
which would be the equivalent of 0.542 pound per 100 pounds of
animal.

On August 18, at 7 a. m., the animal appeared drowsy and sleepy.
At 7.15 a. m. she walked about a lttle, throwing the forelegs out
and dropping them as though lacking control. She staggered a

little as she walked and appeared more sleepy than an hour earlier.

There were some contractions of the surface muscles of the body. —

Plate IT, figure 1, shows the condition of the animal at this time. At
8.15 a. m. she was still upon her feet and able to walk around, but
walked as though tipsy. When standing she appeared sleepy, the
ears drooped, and the eyes were half closed. She stood with her legs
bent at the knees and hocks. At 9.12 a. m. she still stood with the

2 ALT
Po ee

LUPINES AS POISONOUS PLANTS. 23

same sleepy look and hanging ears, and occasionally walked about in
an uneasy way. There was some lack of coordination of the muscles
of the forelegs. Plate II, figure 2, shows the condition of the animal
at that time, and Plate II, figure 3, shows the animal about 9.30 a. m.
At 11.10 a. m. the condition was about the same as at 9.12. At 11.55
a.m. the animal was found upon her knees; she got up with difficulty,
but was barely able to stand upon her feet. She had been gradually
growing easier, and at 12.10 p. m. was down with her head upon the
ground, apparently in a sleep. At this time she was given 5 grams
of tannic acid in water, and 5 minutes later 10 c.c. of gin was given
in an ounce of water. At 12.20 p. m. she was found down in a corner
of the corral with her head under her body. She was helped up, but
fell down again on her side in a convulsive spasm. At 12.25 p. m.
she lay quiet; her respiration was 32, deep and labored, and her pulse
98, strong and regular. At 12.30 p. m. an attempt was made to get
her upon her belly, but she rolled over on the other side. This was
repeated several times. After several attempts she lay upon her
belly with her nose extended on the ground. At 12.38 p. m. she
tried to get upon her feet, but was unable to do so and went over
on her side. Her legs straightened out, her head was thrown back
in a strychninlike spasm, respiration stopped, the eyes rolled back,
’ and at 12.41 p. m. the animal was dead.

At the autopsy a few petechiz were found on the walls of the
ventricles of the heart. The vessels of the small intestines were
injected, and the brain was slightly congested. Nothing else abnor-

mal was noticed.
g TYPICAL CASE OF SHEEP NO. 253.

Sheep No. 253, a ewe weighing 62.5 pounds, was kept in the corrals
August 5, 1914, for feeding.

On August 6, at 9.20 a. m., her temperature was 102° F., her pulse
60, and her respiration 16. From 9.30 a. m. to 10.05 a. m. she was
fed with the balling gun 195.31 grams of seed pods of Lupinus
leucopsis. ‘The pods were collected July 16, before the seeds had
ripened, and allowed to dry. In the process of drying the seeds were
expelled. The pods were ground and 4 ounces of water was added to
facilitate the feeding. This was fed in the ratio of 312 grams of pods
to 100 pounds of animal. Allowing for the loss of weight in the pods
due to drying, this animal received an equivalent of 2.755 pounds of
fresh pods per 100 pounds of animal weight. The sheep appeared
all right during the day and was watched until 11 p. m.

On August 7, at 5.50 a. m., her temperature was 100.6° F., her
pulse was 96, and her respiration 21. The animal was found lying
down with her head through the fence between the boards and swing-
ing it from side to side. Her eyes were staring and expressionless.
The femoral pulse was almost imperceptible. There was no evidence

24 BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.

of dyspnea. At 6.35 a.m. she was still lying upon her belly, with 4
her head through the fence and swinging it from side to side. She
was taken up and put upon her feet and walked a little distance,
staggering as she walked. At 6.40 a. m. her pulse was 140, the heart
beating very hard. Her temperature was 100.6° F. and her respi- .

ration 24. The animal was standing with her head lowered and b |

swinging from side to side. She staggered across the pen, hit her
head against the fence, and pushed forward, with her head partly
twisted and under her body. As she lay down her head kept swinging
from side to side. At 8 a. m. she was able to walk, but staggered as
she moved. Her head was still swinging, pendulum fashion, from
side to side. She pushed up against the side of the pen, preferably
in a vorner, with her héad flexed toward the breast, so that at times
she almost stood upon her head. |

Plate III, figures 1 and 2, taken at 7.55 a. m. and 8 a. m., respec-
tively, show the positions assumed. At 8.55 a. m. the sheep was
lying quietly at the front of the pen. Her respiration was 80. She
no longer swung her head from side to side and did not attempt to
push it against the side of the pen. At 9 a. m. her temperature was
102.6° F., her pulse 102, and her respiration 86. The pulse was
rather weak. There was some dyspnea. She lay on her belly with
her head to the left side and was quiet. At 9.15 a. m. she was up and
about the corral. She would butt against the fence, then fall,

extend the legs rigidly, and breathe rapidly. There was no rhythmic }

motion of the head at this time. At 9.40 a. m. she lay panting, with
some frothing at the mouth. At 9.55 a. m. her pulse was 132, and
strong in the femoral artery. Her respiration was 200, panting; her
temperature was 103.2° F. She got her head into the corner of the
corral and shoved forward until she almost stood upon her head.
She struggled to push herself into this position, and as she was
standing upon boards during the struggle her feet would sometimes
slip and she would fall. After falling she would get up and start the
butting process over again. At 10.35 a. m. she lay with her head
to one side, the respiration being very rapid. She was disturbed,
got up and started butting against the fence as before. This was kept
up for two or three minutes, then she went down and remained lying
with her head on one side. Plate IV, figure 1, shows-her position at
10.40 a.m. At 10.55 a.m. the animal was up again and butting into
the corners as before. She was drenched with 30 c. c. of whisky and
an equal quantity of water. Her pulse was 132 and strong. At
11.55 a. m. she stood with her head low and part of the time on one
side. Plate IV, figure 2, shows her in this position. There was a
rapid chewing motion of the jaws; the motion was dorsiventral and
not lateral. She bumped her head into the fence as before and

PLATE lI.

Bul. 405, U. S. Dept. of Agriculture.

1913.

AuGusT 18,

. M,

BvA

<l

—SHEEP No. 184 AT 7

il

Fic.

1913.

—SHEEP No. 184 AT 8.57 A. M., AUGUST 18,

2

FiG

Bul. 405, U. S. Dept. of Agriculture.

AuGusT 18, 1913

M.,

15 A.

DOT Ais

.—SHEEP No

Fia. 1

TOMS:

J

AUGUST 18

M.,

201 AT 9.12 A.

—SHEEP No.

2

Fic

, 1913.

AuGUST 18

M.,

ZOLA:

AT 9.

201

—SHEEP No.

3.

FIG

&

PLATE III.

1914.

M., AUGUST 7,

253 AT 7.55 A. M., AUGUST 7, 1914.
—SHEEP No. 253 AT 8.00 A.

2

Fia. 1.—SHEEP No.
Fia

Bul. 405, U. S. Dept. of Agriculture.

PLATE IV.

Bul. 405, U. S. Dept. of Agriculture.

Auaust 7 1914

M.,

40 A.

253 AT 10

—SHEEP No.

1

Fia

Aucust 7, 1914

M

152s

253 AT 11

—SHEEP No.

2

Fla.

LUPINES AS POISONOUS PLANTS. 95

staggered when she attempted to walk. At 11.22 a. m. she stood
swaying from side to side. At 11.45 a. m. she was lying upon her
belly with her head on one side, her respiration being about 200. At
11.50 a. m. she was up on her feet and commenced to butt into the
fence again. She soon fell and lay panting. Her respiration was
160. At 11.57 a. m. she was lying upon one side and panting. At
12 noon she suddenly got upon her feet and started butting the
corners, but immediately fell over, holding her head on one side.
The chewing motions, as noted before, were marked. At 12.15 p. m.
she got upon her feet, tried to run but fell over; she picked herself
up, however, and started to butt into the corners. At 12.17 p. m.
she fell down and lay quietly. At 12.30 p. m. she was on her side
struggling in an attack of dyspnea. At this time she was bled by
cutting the angular artery of the eye. At 12.35 p. m. her respiration
| was 160. At 12.40 p. m. the movements of the jaws and lips still
continued. At 1.10 p. m. she was struggling and the muscles of the
flanks were trembling. Her pulse was so fast that it was impossible
to count it. Her temperature was 107.6° F. At 1.11 p. m. she had
fallen over on her side and was moving her body about. At 1.13 p.m.
her respiration was barely perceptible, and at 1.15 p. m. it stopped.
The movements of respiration and of the heart stopped at about the
same time. ,

In the autopsy petechiz were found upon the auricles of the heart.
The lungs were congested. The walls of the ileum were injected, with
some ecchymoses. ‘The mesenteric blood vessels were injected.

LABORATORY EXPERIMENTS WITH EXTRACTS OF LUPINE SEED UPON
MICE.

Preliminary experiments showed that mice are very susceptible to
poisoning by extracts of lupine seed injected intraperitoneally.
Accordingly, a few experiments were undertaken to determine by
this method the relative toxicity of extracts of various parts of the
lupine plant. Seeds, pods, and leaves of Lwpinus leucopsis collected
at Greycliff, Mont., in the summers of 1913 and 1914 were used for
these experiments. The following is a description of the method
used in preparing the extracts:

Material was taken from each part of the plant which it was desired to examine,
and all the samples were ground in the same mill set to the same degree of fineness.
They were all ground the same day and at approximately the same time. From each
form of material a sample of 20 grams was taken, placed in a flask, and macerated
with 150 c. c. of Prolius’s solution for 48 hours. The maceration of the different
samples examined was begun and ended at the same time. Ninety c. c. of Prolius’s
extract was then taken from each flask, filtered, and evaporated down spontaneously.
The residue of each was then extracted with 1 per cent of HCl, using 3 portions.
The solution was filtered off, neutralized with Na,CO,, and made up to 10 c. c., so
that 1 c. c. of the solution was equivalent to 1.2 grams of the sample.

26 BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.

These extracts were made by Mr. O. F. Black, chemical biologist
in the Office of Drug-Plant and Poisonous-Plant Investigations, and —
were used in Be injections upon mice. Table 2 gives
the results.

TABLE 2.—Kelative toxicity to mice of the various parts of Lupinus leucopsis.

| |
Animal. | Plant used. |
| Extract i
Part of plant. used. Per 25 Result.
Mouse. Weight. Weight. | grams of
animal.
Grams CC Grams Grams. |
No7 ae 24 WieaVeSctEL 2.8 Sas o i! 12, 1.25 | Death in 7.5 minutes.
No. 35--- -=-% - 2655 ele ea Qe er Sane aa adn .9 . 833 | Death in 9.5 minutes.
No. 445722. 23 55h |e ARE OG 452 08 5238 eke 45, 6 .64 | Death in 26 minutes.
ING Agios AAR ences Ose eens eee ails -18 . 322 | Not sick.
Infepsieessees 26.5 | Pods (seed removed) ae -6 .565 | Death in 4.5 minutes.
INOS3 fase D0 eilteser ae bak Pcs 2 eee ee - 25 .3 .375 | Deathin 6.5 minutes.
Nps 452 =4o55- 19 =e ni dothe |e sees ms 18 . 237 | Not sick.
INQIS8 2425-2 15.5 ods eed shed) === >= oak <o .482 | Very sick; recovery.
MNos43 222 22 IQSS is32e- Os 32> See eee .3 36 457 | Sick; recovery.
NOs«MbO ee NON ie A Oa eae eee .35 .42 552 | Deathin 11 minutes.
NoI39 Ss 24 aaa cists collection) 22 . 24 -25 | Death in 12.5 minutes.
INOSA0S 2235-2 19a ES OOl eee ere eee -15 - 18 . 237 | Sick; recovery.
INOsAI 25 122. - 22.5 | Seed Lists collection) LZ . 24 .266 | Not sick.
IN ONED Eres DAs ASS OR as ae pene a ae -20 .3 .357 | Death in 14 minutes.

Table 2 shows that the amount necessary to kill a mouse on the
basis of the standard weight of 25 grams is approximately as follows:

Grams.
TG OAV CBE see 2 a ec ae eB ee tener 0. 6 to 0. 64
iPods (seed removed) os = * 522. 25 kien ee ees eee as eFC gees 1s
Pods(seed shed). oe orcs Se eic te ee ee eae a .5
Seeds. coligetion OL AOI 3s. i eee es ee eee SAFI a
Seeds, collection: of Vue eee eee oe a a ee een ae . 25

Inasmuch as it is known from other experiments that the toxic and
lethal doses are practically the same, it is fair to assume that this
table gives, approximately, not only the lethal dose but also the
toxic dose. From Table 2 it appears that the seeds collected in
1914 were slightly more toxic than those collected in the preceding
year. Possibly a certain amount of the toxicity was lost in keeping.
It appears also that the pods from which the seed had been shed
were only about half as toxic as the seed. The pods from which the
seed had been removed were considerably more toxic than the pods
from which the seed had been shed, and the leaves were only about
one-half to one-third as toxic as the seed. These figures do not con-
form very closely to those obtained in the field experiments upon
sheep, but perhaps as closely as could be expected under the differ-
ent conditions of experimentation. An especially interesting feature
of this experiment was the definite proof obtained by Mr. Black by
analytical methods of the presence of the alkaloids in the leaves,
with the consequent deduction, confirmed by actual experiments,

' LUPINES AS POISONOUS PLANTS. Dil

that the leaves are toxic. Inasmuch as the field experiments upon
sheep were inconclusive in regard to the toxicity of the leaves, this
is an important addition to the knowledge of the subject.

The field experiments gave little information as to the relative
toxicity of the different species of lupimes. There was nothing to
indicate any difference in the toxic effect of those used in the feeding
experiments. Accordingly, afew experiments were made on mice with
extracts of the seed of the available species. Seeds of three Ameri-
can species were used, and through the kindness of Mr. Piper, of the
United States Department of Agriculture, seeds were obtained of the
three common European species, Lupinus albus, L. angustifolius, and
L. luteus. - Extracts of these lupines were prepared by Mr. Black, using
exactly the same method as that employed in the preceding test of
the seeds of L. leucopsis. A series of tests was made on mice by
intraperitoneal injection of these extracts. Table 3 shows the re-
sults. In this table, for purposes of comparison, the experiments
with seeds in Table 1 are included.

TaBLE 3.—Comparative toxicity to mice of the seed of different species cf lupines.

Used per 25 grams
of animal.
Plant. Result.

Mouse. | Weight. |} Extract.| Plant. | Extract.| Plant.

Animal. Amount used.

Lupinus leucopsis: Grams. Cxe: Grams. Cres Grams.

1914 seed. .-.-..- No. 39. 24 0. 20 OR24 ee oak 0. 25 Death in 124 minutes.
ID Ou eae eas. No. 40- 19 o15 AL SAE Peels .237 | Sick; recovery.
IGISISce Cea acee: No. 41. 2255 - 20 SAN SIREN .266 | Not sick.
MOees 5258 No. 42. 21 ~20 Om Meee ees 357 +| Death in 14 minutes.
Lupinus luteus...-.- No. 49. 18.5 ald .18 0.2 2D None.
IDOE auc eee ee INOSA9S Eee ew 35 -6 ‘il -8 Sick; recovery.
IDO Hee ese cece No. 50- 21 B20, 3 3 - 36 Symptoms.
Lupinus albus. ..--- No. 51. 23 -5 6 - 54 . 65 Death in 3 minutes.
DOA ost ee No. 52- 23.5 2 - 24 21 325 Death in 13 minutes.
pups angusti- | No. 53. 24 25 3 - 26 23 Death in 19 minutes.
olius.
DOs ee Sree No. 54. 27 215 18 14 17 Symptoms.
Hupinus leucophyl-
us:
1913 seed....... No. 56. 2255 «25 Sere Gaeaaenacd -335 | Death in 64 minutes.
Wot si wes: No. 57. 21.5 215 SSM aes Seise - 2075 | Symptoms.
Lupinus argenteus:
1913 seed ....--- No. 58- 21.5 <20 POR eee cece -35 Death in 4 minutes.
1D OVS arene No. 59. 26. 75 calls Las meray .168 | Death in 6 minutes.
Dorie sts3: No. 60- 19 15 oO Os eeereee ane . 1185 | Symptoms.

In regard to mouse No. 49, it should be stated that inasmuch as the
first injection of 0.15 c.c. of extract produced no results, a second in-
jection of 0.5 c.c. was given shortly afterward. From the known facts
in regard to lupine poisoning it is not to be presumed that there was
any appreciable accumulative effect from the preceding injection.
Table 3 shows very clearly that by this method of experimentation
Lupinus albus and L. angustifolius appear to be about equally toxic.
L. luteus, however, is much less toxic, it being necessary to take prob-
ably three or four times the quantity in order to produce results. It

28 BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.

may be noted in passing that L. luteus is the species which is said to
be responsible for the so-called ictrogenic poisoning which occurs in
northern Germany. In regard to the American species, L. lewcopsis
and L. leucophyllus are about equally toxic and do not differ much

from L. albus and L. angustifolius. L. argenteus appears to be much’

more toxic than the other American species examined.

It should be noted that in these experiments on mice the animals
died of respiratory paralysis, with marked dyspnea, the heart some-
times continuing to beat as much as a minute after respiration had
ceased. The work with extracts is of a preliminary character, and
it is intended to prepare for future publication an extended study of
the alkaloids and the effect of the extracts.

PART III.—DISCUSSION AND GENERAL CONCLUSIONS.
FAILURE TO POISON SHEEP IN 1910, 1911, AND 1912.

In the early work of the field experiments it was assumed that the
lupines were sufficiently toxic to poison animals in the course of ordi-
nary feeding. It was thought that if the animals were confined and
given little or nothing of other food material and were abundantly
supplied with lupine, symptons would appear if the plants were toxic.

There was little in American literature to indicate the probable
dosage. Chesnut and Wilcox (1901, p. 108) state that 150 pods were
fed to each of two sheep and both were fatally poisoned. Accord-
ingly, in 1910 the sheep used were confined in corrals and were fed all
they would eat. One animal, sheep No. 102, between July 6 and 30,
ate 128 pounds of lupine tops, including leaves, flowers, and fruit, and
another, between August 25 and September 7, ate 55.5 pounds of
seed-containing pods, with no ill effects other than a loss of weight.
Sheep No. 105, which received the 55.5 pounds of pods and seeds, was
given on some days as much as 8 pounds.

The experience of 1911 was similar and with no more results.
Although a large number of experiments were made in 1912, in only
one case were there harmful results. This animal, sheep No. 180,
was fed 0.85 pound of lupine seeds on September 15, and the next
day it was found dead in the pasture. At that time, because of the
large number of negative experiments, it was thought that the death
of the animal must be due to some other cause than lupine poisoning.
The more complete knowledge of the effect of lupine, brought out in
the work of 1913 and 1914, makes it evident that this was a case of
lupine poisoning. All the other cases of 1912 were negative.

In comparing the work of these years it will be noticed that in
1913 the feeding of seeds was done in a single day, and in most cases
the quantity fed was eaten in a short time. This was true of sheep
No. 180 in 1912, which ate the given quantity within an hour. In

LUPINES AS POISONOUS PLANTS. 29

1914 the experiments were by forced feeding, so that the animal
received the total quantity in a short period of time; while in 1910
and 1911 some of the animals received large quantities, but the feed-
ing was distributed over a long period. Although in some cases as
much as 8 pounds was fed in a single day, this was eaten in a more
or less leisurely way.

In 1912 two sheep, Nos. 175 and 168, were drenched with the fruit,
No. 175 receiving 1.435 pounds and No. 168 4.198 pounds, and
showed no symptoms, although the 1914 experiments indicated that
the toxic dose of fruit is about 14 pounds. Sheep No. 175, however,
received its lupine in three doses, between 10.30 a. m. and 3.30 p. m.;
it is possible that this animal might have been poisoned had it re-
ceived the material in a single dose, as the quantity given was close
to the toxic limit. But sheep No. 168 received 4.198 pounds, be-
tween two and three times the toxic dose as determined in 1914;
this material was given in six doses, between 8 a.m. and 7 p.m. It
seems probable that the only reason this animal was not poisoned
was because of the length of time during which the material was
given.

It appears to be a fair inference that the excretory apparatus of
the sheep can take care of the toxic substance of the lupine provided
the quantity given at any one time does not reach the toxic limit.
It is hoped to carry on later detailed experiments to show just how
this is done—experiments for which adequate facilities have not been
available thus far. It is probable, however, that this excretory work
is done largely by the kidneys, since preliminary experiments with
other toxic substances upon sheep indicate that the kidneys are
very efficient in the removal of some toxic substances. It has also
been shown by others that the lupine alkaloids are found in the
urine. The failure to get results in the earlier experiments was due
to the fact that the sheep did not get at any time more than the
excretory apparatus could remove before the toxic limit was reached.

LUPINE NOT A CUMULATIVE POISON.

The lupines as poisonous plants do not have a cumulative effect.
This has an important bearing on range management of sheep, for
it is evident that sheep may graze continuously on lupines with no
bad results, provided the toxic limit is not reached at any one time,
Inasmuch as the toxic dose is a fairly large quantity, and sheep do
not ordinarily show any special fondness for the lupmes, well-fed
animals are not likely to be poisoned by lupines. It is only when,
for some reason, they eat an unusually large quantity that losses
occur. This is discussed in more detail under ‘‘ Range conditions
under which sheep are poisoned” on page 39.

30 BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.

TOXICITY OF DIFFERENT SPECIES OF LUPINE.

The field experiments give little information as to any difference
in toxicity between the species of lupmes. While only one of the
two species used in the experiments with horses produced effects, viz,
Lupinus myrianthus, the feedings of L. comatus were not conducted |
under exactly the same conditions, the period of feeding being con-

siderably longer than in the experiments in which sane was pro-
duced by L. myrianthus.

The probable reasons why sheep were not aeiabaca on Lupinus
myrianthus and L. comatus are discussed on page 28. Nearly all the
experiments at Greycliff were on L. leucopsis. The field experi-
ments on L. argenteus and L. leucophyllus were so few that it is not
safe to make any deductions. It seems probable that in the experi-
ments of feeding the seed of L. argenteus, the dosage was just a little
too small, even granted that the plant is equally toxic with L.
leucopsis.

The experiments with mice detailed on pages 25 to 28 indicate that
the European species, Lupinus albus and L. angustifolius, are about
equally toxic with L. leucopsis and L. leucophyllus, while L. luteus is
much less toxic, the dosage probably being at least three times as
great as that for the other species. Inasmuch as the experiments
on mice were few in number, too much importance must not be at-
tached to the results. They are interesting, however, as giving some
indication of the probable comparative toxicity of the species ex-
amined. These experiments with mice give an indication of the
probable toxicity of the leaves as compared with other parts of the
plants.

TOXICITY OF LUPINE LEAVES FOR SHEEP.

Sheep No. 243 was an undoubted case of lupine poisoning. The
symptoms were typical and it received only leaves. It was impos-
sible, however, to verify this by other experiments. The other ani-
mals (Nos. 295, 266, 256, 290, and 306, with L. leucopsis, and Nos. 263
and 307 with L. argenteus) were fed much more, in some cases more
than three times as much, without any effect whatever. It should
be added that while all animals upon the range apparently graze
upon lupine with impunity, sometimes eating very large quantities,
there are cases when it seems probable that lupine leaves must be
toxic. For example, cases of undoubted lupine poisoning in the
Absaroka National Forest, examined by Dr. Hadleigh Marsh in the
summer of 1914, apparently did not have an opportunity to obtain
enough of the seeds to cause the difficulty. If it could be assumed
that the leaves are sometimes toxic, these cases would be much more
easily explained. Many similar cases of range poisoning have oc-
curred where it was difficult to explain the loss except by assuming

LUPINES AS POISONOUS PLANTS. al

that the lupine leaves are toxic. Moreover, laboratory work, con-
ducted with the assistance of Mr. O. F’. Black, shows clearly that there
is a large quantity of the alkaloid in the leaves, although less than in
the seed.

A careful analysis of the cases of sheep fed with leaves offers no ex-
planation for the difference in results, although many possible factors
have been taken into consideration, such as difference in animals, differ-
ence in methods of administration, seasonal change of toxicity, ete.
All that can be said at this stage of the investigation is that while lupine
leaves are not always injurious they are toxic, and under some condi-
tions—conditions which can not now be defined—they cause illness
and death. It is evident that no definite statement can be made as
to the toxic and lethal dose of leaves for sheep.

TOXICITY OF LUPINE SEED FOR SHEEP.

Table 4 is astatement of the sheep poisoned by lupine seed in 1913,
showing the quantity in each case which produced the result.

TaBLE 4.—Sheep poisoned by lupine seed in 1913.

| Pounds

Mert fed per £
Feed and animal. Date fed. 100 pounds Result.
of animal.

Unground seed:

INO? ODS 36 See oe an oe OS a = aeRO eS Aug. 15 0.561 | Sick; recovery.

INIOs US GES Sea Ree oS iss Se eee ee ee Ie BN Raa Aug. 17 705 eath.
Ground seed:

Oj, FAD) 4 ee SS SEe es ie Se Sata ve SPN eS Aug. 15 . 268 | Sick; recovery.

INO Ze oe cae oc occ coc obo dauae peek ooe See een ee Sareea ease Aug. 17 . 542 eath.

Note ae eer rn er ee ee” Aug. 31 441 Do.

INR De RE ee OC eat a Bah a am SI Set Sept. 2 . 245 | Sick; recovery.

ING MBs csosaodaces sense Gul see SO ee ai epeee tens eae Sept. 6 439 Do.

Inasmuch as sheep No. 208 received 0.551 pounds of the unground
seed without any toxic effect, it would appear that the dose received
by sheep No. 209 (0.561 pound) with resulting illness, must have been
very hear the toxic limit. As the only sheep that died (sheep No. 184)
received 0.705 pound, the lethal limit, so far as these experiments
show, lies between 0.561 and 0.705 pound.

When ground seed was used, the dosage was much smaller, as
would be expected. Sheep No. 212 was poisoned by 0.245 pound,
and sheep No. 210 by 0.268 pound. Others were fed larger quantities
without effect; for example, sheep No. 196 received 0.425 pound, and
sheep No. 207, 0.423 pound, while several received 0.3 pound or
more. Sheep No. 196, however, received its dose in 2 days, whereas
the others received theirs in a single day. It seems that sheep may be
poisoned on as little as 0.25 pound or may receive as much as 0.423
pound without effect.

Syp4 BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.

The two cases of death resulted from doses of 0.543 and 0.441 i
pound. The general deduction from the experiments of 1913 on |
ground seed was that the toxic dose was between 0.25 and 0.5 pound,
and the lethal dose about 0.5 pound, with the probability that in the —
average cases the lethal dose was somewhat more, perhaps nearly 0.6 |
pound. i‘

It was considered that these experiments determined the dosage
nearly enough for practical purposes, and in the experiments of
administering ground seed in 1914, which were mainly with reference
to the effect of antidotes, the dosage was intended to be heavy enough
to produce serious illness, if not death. The general results showed
that the estimate of dosage made in 1913 was approximately correct,
and that the toxic dosé is somewhat less than 0.5 pound, sometimes
as low as 0.25 pound.

TOXICITY OF LUPINE PODS FOR SHEEP.

The number of experiments of feeding lupine pods was not suffi-
cient to determine the dosage with any exactness. Sheep No. 253
died on August 6, 1914, from eating 2.755 pounds of pods. This
material had been collected previously and dried, but the weight as
given is the green weight, as the loss from evaporation was known,
These were pods in which the seed was not yet ripe.

Sheep No. 301 died on August 28, 1914, from eating 2.041 pounds
of pods which were collected after they had shed most of their seed;
a few seeds, however, remained attached to the pods. The weight
given is the dry weight, as there was no way of estimating the loss
of water in drying. Im all the other cases much smaller quantities
were fed, and no intoxication resulted. It can only be said that the
pods are distinctly toxic, but the dosage is much greater than of the
seeds.

It may be added in this connection that in 1913 a careful compu-
tation was made of the relative weights of the seeds and pods in the
fruit, and that, based on the result of this work, the toxic dose of pods
would be 3.4 pounds. Without much doubt the toxicity of the pods
varies at different seasons and probably is much reduced in the dried
pods remaining attached to the plants in the late summer and fall.

TOXICITY OF LUPINE FRUIT FOR SHEEP.

By ‘“‘fruit’’ is understood the pods with the contaimed seeds. A
considerable number of experiments were made to determine the
toxicity of the fruit as compared with the seed. Table 5 gives the |
results of these experiments. The term ‘‘Seed heads’’ means the |
fruits and the stems bearing them. In the cases listed under “‘ Fruit,
fully developed,” the pods were picked from the stems.

The last four feedings under ‘‘Fruit, fully developed” were of:
very poor, locoed animals; consequently, the dosage (as computed

LUPINES AS POISONOUS PLANTS. Be

according to weight) would be distinctly different from that com-
‘puted for the same animals in a normal, healthy condition. This
may possibly explain why in the cases of Nos. 251 and 280 the dosage,
computed on the basis of a 100-pound animal, so much exceeds the
lethal dose shown in the preceding experiments. For example,
sheep No. 280 weighed 99 pounds earlier in the season, while at the
time of the experiment it weighed only 84 pounds. If the dosage of
material given September 4 were computed on the assumption that
the animal had its original weight, it would reduce the amount given
in the table to approximately the toxic or lethal dose of the preceding
animals, and as the margin between no symptoms and toxicity is so
small this animal would not differ materially from the others.

TABLE 5.—Sheep given forced feedings of lupine fruit in 1914.

Pounds fed

Feed and animal. Date fed. Saat Result.
animal.
Fruit, fully developed:
Os, Ae Soi ae ea a ae a ily Gseeee ss eae 0.882 | Not sick
INOS Plt 3s BBs as As oN NES es aaa gee ea AOU aaa ee ee 441 (0)
INO BUC Sie Se eS hist Cees Ra et eee eon anes ie anee dibuhy ise sepeasae 1.322 Do
INOS ZIG GAS SNe ASA ates SH OC Rep reas ca ere DaTliylO eee esi 1.764 | Death
INO 2 5 Sie ese ee eee ee See eee eS Duliy, 20s eens oe 1.548 Do.
INOS2 OO eGe e mama me Ny ee eke dhol hy PB Aoseeeauemrs 1.433 | Not sick.
INOS 2A 0 eM secon ye es ideas es ce ery Soe poe ot July 2ee ee see 1.543 | Symptoms.
INO 29a eae Seer seen. ves LeU eRe esi Peers COzetaee sees. 1.433 | Not sick.
ING SAB OAS 5 Sac aa Sasa Bok Aaa Ses es ee ditty Paes ba SeRaasse 1.763 | Death.
ING@y PRs Ss oS be Bae a Ae ree eee ine ee ie Onan NG Feds GAY eee ere oe 1.543 | Not sick.
IND: DES ae ee Es a a eee I Nib ae es eee 1.541 Do.
INOW 25 Rina Sepia he eas See Se ee as Lobe oe I Sep te 2ieseee ene 1.761 Do.
Ob PD Sc BONE ae ae ee ee a ee ee Sept. 4 e ss ee 1.901 Do.
Seed heads, fully developed:
ING PND OS Sosa oad ee are emt e eenremasae ate liys28=29 =e eee 1.033 Do.

It is evident from Table 5 that approximately 14 pounds of fully
| developed fruit will produce symptoms or death in a 100-pound
_ sheep—that is, it takes three times as much of the fruit to poison as
of the seed.

SYMPTOMS. :

Some of the sheep poisoned by lupine, froth at the mouth, but this
is by no means a universal symptom. The most noticeable and
significant symptom is the character of the breathing. In the milder
cases the breathing is heavy and labored, subsiding into a condition
of coma in which the animal may continue for a long time, snoring
as though in a deep sleep. If able to stand, the animal may fall
over in its sleep. In the more acute cases, there are severe attacks
of dyspnea, during which the animal throws itself about violently
in its attempts to breathe. During these attacks the tongue and
mouth become cyanotic from the congested peripheral blood ves-
sels. Sometimes in these attacks of dyspnea the animal dies in
convulsions in which the limbs are extended rigidly, much as when
poisoned by strychnin. In other cases the condition of coma deep-

84 BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.

ens until the animal dies without a struggle.
of dyspnoea, however, may be considered typical of lupine poisoning,

The convulsive attacks

Drooping of the ears is noticeable in the early stages of the poisoning, ©

In many cases the poisoned animal is continually pushing its head

|

against surrounding objects. In corral cases the sheep pushes —

against the fence, lowering its head so that sometimes the animal |
almost stands on its head. These attitudes are shown in Plate Iil@
figures 1 and 2. If moved from one point it may immediately push _

up against the fence in another place, sometimes throwing its head

A./7.

LE pee LEN DSS leslie ail a aa
EMO Seema meer:
tee) BBE Mis
ENE | ose 1 ae ae Le

et | eeparazroy | | |

through an opening |
and remaining in that —

position. Sometimes

the

from side to side.

Generally when ©
sheep become af-_
fected on the range —
they run about ina
frenzied way, butting —

into otheranimalsand

objects. Handling the ~
animals under such —
excites

conditions

them to such an ex-

aes
Minas iielaiab telat ar
ore
nue

Sere eee ee ar
PLN | | | pancecmarne

interfere with their

remedies by drench-

the animals lost more
Fic. 2.—Curves showing the changes in pulse, respiration, and tem-
perature of sheep No. 185.

gained from the remedy. If the sheep does not die in the period of

excitement, it staggers until it falls, then lies in a stupor, which in |

the fatal cases gradually grows deeper.

The pulse and respiration are very high in the acute stages of the ~

intoxication, but later are not very different from normal, except
that the pulse rate is frequently increased just before death.
The intoxication produces no effect on the temperature, but in long-

continued cases the temperature gradually falls, sometimes to be- _
tween 98° and 99° F. The curves (fig. 2) showing the changes in —

head will be —
swung rhythmically —

tent as seriously to —
chances of recovery. —
When attempts were —
made to administer —

ing, it was found that —

by the increased ex- —
citement than they —

LUPINES AS POISONOUS PLANTS. 35

the rate of the pulse, respiration, and temperature of sheep No. 185

are fairly typical of the changes which ordinarily take place.

Associated with the condition of excitement there is frequently
more or less violent trembling. Nausea is not often exhibited.
Bloating occurs in some cases, probably being more likely to occur
when the sheep eat very largely of lupine leaves.

Especially characteristic of lupine poisoning are: (1) Excitement,
leading to running about and butting into other objects; (2) con-
vulsions, accompanying the attacks of dyspnea; and (3) heavy breath-
ing, sometimes accompanied by snoring.

There is considerable variation in the length of time which elapses
between the feeding of lupine and the incidence of symptoms of poison-
ing. In some cases symptoms appeared in 1 or 2 hours, while there
were other cases in which nearly 24 hours passed before symptoms
were noticed. It is evident that rumination does not necessarily
precede the symptoms; in fact, there was no evidence that rumination
occurred in any of the experimental sheep, although as they were not
under constant observation it could not be said positively that rumi-
nation never took place.

It can be seen from the experimental work that sheep upon the
range, when poisoned by lupine, may not exhibit symptoms until
they have left the source of their trouble far behind. —

In the autopsies, the peripheral blood vessels were found strongly
congested. The left heart was usually strongly contracted, and upon
the surface of the heart in most cases were petechiz. The lungs were
congested, and sometimes the liver. The blood vessels of the brain
were somewhat congested, and generally the blood vessels of the inner
wall of the ileum were more or less congested.

The immediate cause of death is evidently respiratory paralysis.
Death may follow very quickly after the first symptoms, or the ani-
mals may live for 2 or 3 days.

As noted in the description of the experiments of intraperitoneal
injection of lupine extracts in mice, the animals died of respiratory
paralysis, showing symptoms which were comparable with those ex-
hibited by sheep.

The preceding details of symptoms are those exhibited by sheep.
The experiments with horses were so few that no complete picture
can be given. The distinct symptoms noted were twitching of the
surface muscles, constipation, dullness, and a tendency when walking
to lift the fore feet high.

Dr. A. D. Knowles, of Butte, Mont., has recently treated some
interesting cases of lupine poisoning of horses. Dr. Knowles has
made a very careful study of these cases and has done some experi-
mental feeding, making autopsies and having microscopic prepara-
tions made of the diseased organs. ‘These horses exhibited symptoms

36 BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.

comparable with the “‘ictrogenic” poisoning reported in Germany,

especially noticeable being the atrophy of the liver accompanied by

a yellowing of the connective tissues.

While some American authors have distinguished between ‘‘acute”
and ‘‘chronic” lupine poisoning, their statements appear to have
been drawn from European sources, and Dr. Knowles has apparently
been the first to note and record definitely cases which can be diag-
nosed as instances of ‘‘lupinosis”’ or ictrogenic poisoning. The clear-
cut evidence presented by Dr. Knowles seems conclusive and makes
it probable that other similar cases will be found, although it does
not seem likely that lupime causes large losses of horses in the ag-

gregate.
PATHOLOGY.

Some typical pathological material from the autopsies was exam-
ined by Dr. Formad, of the Bureau of Animal Industry. A large
amount of material remains for examination, which may be made
the subject of a future report. The general character of the pre-
liminary report was confirmatory of the results of the macroscopic
examination of the cases.

There was fullness and congestion of the pulmonary capillaries.

The hepatic cells showed the presence of a mild degree of cloudy
swelling and some cedema. ac!

The kidneys showed a slight degree of cloudy swelling of the
epithelium, and in some cases fullness between the convoluted
tubules of the cortex and overdistension of the capillaries of the
medulla accompanied in places by outwandering of the leucocytes
and diapedesis of the red-blood corpuscles, indicatmg a marked
congestion.

COMPARISON OF “LUPINOSIS” AND POISONING OF SHEEP BY
AMERICAN LUPINES.

The symptoms of lupinosis are described in detail by a number of
European authors, e. g., Ziirn (1879), Arnold and Schneidemihl
(1883), Roloff (1883). Summaries are given by Dammann (1902, p.
340-341) and Hutyra and Marek (1913, p. 524-525). |

The characteristic symptoms are:

(a) Loss of appetite.

(b) Fever in the first stages of the disease.

(c) Weakness.

(d) Cerebral excitement; thrusting the head against a wall and into corners.

(e) Gnashing of teeth.

(f) Pain in hind part of body.

(g) Diarrhea; ill-smelling excrement.

(h) Sometimes bloody urine, containing bile and albumen.

(<) Yellowing of conjunctiva and visible parts of mucous membranes in most cases,
but not in all.

(7) In some cases swelling of ears, eyelids, lips, and nose.

eh wan ahh

ee ie

a

LUPINES AS POISONOUS PLANTS. 37

(k) In autopsies there is seen a citron-yellow color of the body tissues, hemorrhages

_ in various parts of the body, especially in the mucous membranes of the alimentary

canal, the peritoneum, omentum, mesentery, the epicardium and endocardium,
frequently fullness of gall bladder, generally an acute yellow liver atrophy, while
in other cases the whole picture presents an appearance of acute phosphorus poisoning.

The German authors distinguish between acute and chronic
cases, the symptoms being the same, but in the latter the liver
exhibits chronic interstitial inflammation, leading to atrophy of the
organ, this being accompanied by nephritis and enlargement of the
spleen. |

It will be noticed that the laboratory results obtained by Sollmann
and by Clawson and Black agree very well with the field experiments
with sheep, but differ very distinctly from the symptoms of lupinosis.
The fever and jaundice which are especially characteristic of lupi-
nosis have never been observed in sheep in America, either in experi-
mental feeding or in poisoning upon the range. It must be con-
cluded that ictrogen is not the cause of loss of sheep on the ranges of
the United States, but that the poisoning is due to the alkaloids in
the lupines.

The symptoms in the corral experiments and in range cases are
distinctly those of alkaloidal poisoning. The question naturally
arises as to the explanation of the difference between the poisoning
of animals as exhibited in Germany and in the United States. The
lupines examined in Germany possess alkaloids which are similar to
those found in the American lupines, if not identical with them, and
yet few clear cases of alkaloidal poisoning of domestic animals have
been reported, while in America there has been no poisoning from
ictrogen. In the absence of any determination of what ictrogen
really is or how itis formed, only a hypothetical explanation can be
given. If it is granted that ictrogen is the cause of lupinosis and if
the opinion is accepted, which seems to be held by the later authors,
that ictrogen is formed by the action of microorganisms upon the
lupine, a possible explanation lies in the different conditions of the
countries. The European lupines are cultivated plants, grown and
handled like hay. The poisoning cases are caused by lupine that is
exposed in the mass, and sometimes under conditions favorable to
the growth of microorganisms. The American lupines are wild

plants, which grow in a somewhat scattered manner; they are not

collected in masses, and consequently do not have an especially
favorable environment for the growth of microorganisms. More-

over, it is very possible that the particular organisms which produce

the toxic substance in Europe are not present in this country; of
this nothing can be said positively, for no one has yet been able to
demonstrate that any specific organism or group of organisms is
responsible for the hypothetical substance ictrogen. Therefore, on
the supposition that there is such a substance as ictrogen, or lupino-

38 BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.

toxin, it seems possible that it would not be produced in America, |
at least under range conditions.

The fact that there is so little evidence that domestic animals in
Europe suffer from the alkaloids of lupine may be explained by the
different way in which the animals are handled. It is shown else-
where in this paper that lupine poisoning of sheep on the western
ranges is ordinarily associated with deficiency in the food supply. —
Rarely, if ever, are well-fed sheep poisoned. It may be assumed
that the sheep in Germany that feed upon the lupine are stall-fed or
pasture-fed and eat somewhat at leisure, so that seldom would they
be likely to reach the toxic limit in a single feeding. In this connec-
tion, however, it may be noted that some of the symptoms men-
tioned by the German authors resemble those of alkaloidal poisoning,
and, granted that ictrogen is the principal cause of the losses, it is
possible that there is a certain amount of alkaloidal poisoning in
addition. This would account for the nervous symptoms described
by the German authors, for these correspond to the phenomena
exhibited by western sheep suffering from the lupine alkaloids.

It should be noted in this connection that the work of Dr. Knowles
mentioned on page 35 seems to show that under some conditions
ictrogenic poisoning of horses may occur in America.

The exact relationship between poisoning by lupine alkaloids and
ictrogenic poisoning is far from clear, and it 1s to be expected that the
chemical investigations which are now being carried on by the United
States Department of Agriculture will aid in clearing up this subject.

REMEDIES.

It was hoped that some remedy might be found by which recovery
from lupine poisoning might be aided, but the results of the experi-
mental work in this direction were disappointing. Whisky, gin, and
atropin were tried, with no beneficial results. Caffein and Epsom
salts were used to increase excretion, but without any advantage.
Tannic acid and sodium bicarbonate were used as antidotes. In the
publication on Zygadenus,' there is a discussion of reasons why the
use of antidotes in a drench in any ordinary method of administra-
tion should not be expected to be beneficial. As in the work on
Zygadenus, experiments were made of giving sodium bicarbonate in
frequently repeated doses. This was tried in four cases (sheep Nos. _
297, 254, 296, and 269). Sheep No. 269 received 0.772 pound of ©
seed and died in spite of the remedy, but sheep No. 296 received
0.666 pound, considerably more than the toxic dose, and was not
sick. This animal received the sodium bicarbonate every half hour,
while sheep No. 269 received it at hour intervals. There seems to be
little doubt, as in Zygadenus poisoning, that if the sodium bicarbonate

1U.S. Dept. of Agr. Bul. No. 125, p. 41.

LUPINES AS POISONOUS PLANTS. 39

can be administered at intervals frequent enough to catch the toxic
principle as it enters the fourth stomach, recovery may be aided.
This is of considerable theoretical interest, and the method might
| be used in order to save an especially valuable animal, but, of course,
range animals can not be treated in this way, for the herder may
have a large number of sheep sick at the same time. It is not
unusual for 200 or 300 cases to occur suddenly, and any remedy
| which involves the administration of more than one or two doses is
useless.

Herders frequently bleed sheep poisoned by lupine and claim good
results. There seems no logical reason for this, however, and the
experience of the writers at the field station leads them to consider
bleeding as harmful rather than beneficial.

RANGE CONDITIONS UNDER WHICH SHEEP ARE POISONED.

It must be recognized that corral experiments, while superior to
laboratory work in unraveling the complex problems of plant poison-
ing of domestic animals, do not cover the conditions of range poison-
ing. Itissometimes very difficult to decide to what extent the results
of intensive corral feeding can be used in the explanation of range
| phenomena. Laboratory and corral experimentation are, of course,
_ essential in such a study, and may give, in fact do give, fundamental
information. An intimate acquaintance with range conditions is,
however, necessary for the practical elucidation of these problems.
Such an acquaintance with range conditions is somewhat difficult for
| the scientific investigator to acquire. Sheep are grazed in locations
| situated at long distances from regular avenues of travel or communi-
| cation, and they are also moving from place to place. Cases of
poisoning are reported, perhaps, some days after the trouble has
occurred, and investigation at that time is likely to be useless. It is
very difficult for the investigator so to locate himself as to be able to
see these cases when they occur. These difficulties doubtless explain
why there has been so little exact knowledge of the conditions sur-
rounding the losses. There has been, moreover, no way of placing a
_ correct valuation on the reports made by sheep herders and owners.
| The average sheep herder does not have a high order of intelligence,
and this has led to an underestimate of the testimony of these men,
for the fact has been overlooked that many of them are keen observers
with a practical knowledge of conditions far superior to that of the
average scientific investigator. The writers of this paper have had
long and intimate acquaintance with the western stock ranges, but
they have frequently been put to shame by the wonderful, almost
instinctive, readiness with which an experienced sheepman will
unerringly recognize slight symptoms of disease in the members of
his flocks. The writers were fortunate in being able to observe many

40 BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.

range cases, and the conditions under which poisoning may occur
seem now quite clear.

It was early recognized that lupme poisoning ordinarily occurs |
only when hungry sheep graze upon the plants. Well-fed bands —

are rarely, if ever, poisoned. This statement, it may be said in

passing, can be made general and covers practically all poisonous |

plants. If sheep are taken from the cars and turned into a pasture
abundantly suppled with lupine, disastrous results are likely to

occur. In July, 1914, 400 sheep, out of a band of 4,000, were lost |

near Lakeview, Oreg. The matter was investigated by Mr. Norman

G. Jacobson, of the Forest Service, who found that the sheep had

been driven 34 miles in four days with a scarcity of forage. On |

July 25, after a day’s drive of 10 miles, they were turned into a 10-
acre pasture which, investigation showed, contained little but sage-
brush and lupines, and the lupines were in pod. The sheep, of course,
ate the lupines and with resulting loss. Many losses have occurred
in the fall when sheep are coming down from the mountain ranges
and have been caught by snow. On such occasions the fall of snow
may cover the grasses, but leave the lupines exposed. Hungry sheep
coming upon such an area may eat enough of the lupines to produce
poisoning. In the fall of 1913 a sheep owner in Montana lost 300
sheep in this way.

An area in the Caribou National Forest was investigated where
annual losses have occurred. It was found that the sheep coming
- from the mountains pass through thick patches of lupine and eat
it greedily. The fact that the lupines are in pod at the time of the
drive makes the matter worse. In 1911, an especially disastrous year,
one outfit lost 1,000 head in this locality.

And yet, in spite of these known cases of severe loss, sheep some- 7

times graze on lupine through a good part of the season and with no
harm. Except as they are especially hungry sheep rarely, if ever,
eat enough of the lupime to cause trouble. Poisoning is much more
likely to occur if sheep are hurried over a lupine area, for then in
their eagerness and jealousy of each other they seize upon the lupines,
which are more easily reached than the grasses. Generally speaking,
it is much better to drift sheep over a lupine area than to drive them.
It may be noted that sheep that are new to a locality are much more
likely to eat too much of the lupine than those that are accustomed
to the country.

An attempt was made in 1912 to demonstrate in an experimental

way that hungry sheep may be poisoned when they feed upon lupme.
A bunch of 11 sheep were kept without food for 36 hours and then,
during one day, were driven 12 to 15 miles with very little opportu-
nity to eat. About 5 p.m. they were brought to a thick lupine patch
and allowed to graze. They fed until about 8.30 p. m., when they

LUPINES AS POISONOUS PLANTS. 41

were bedded down. They were up and feeding again at 12.15 a. m.
and were grazing most of the time until 6.30 a. m., with the excep-
tion of the hour from 4 to 5. To the disappointment of the observ-

ers none of the animals was poisoned. It was noticed, however,

that while a good deal of lupine was eaten they did not take to it
greedily and preferred the grass, which was in fair abundance.

In thinking over the experiment later, it seemed probable that the
fact that the sheep were allowed to feed freely, combined with the
abundance of grass, might explain the lack of results. It was there-
fore decided to repeat the experiment with the difference that the
sheep should not be allowed to feed freely upon a lupine patch, but

_ should be kept moving back and forth, the idea being that in this

way they might snatch at the lupine as the most prominent plant.
An experiment of this kind was tried July 31, 1914. Thirty-six

| sheep were corralled at night and kept in until 2 p.m. the next day

with no food. At 2 p.m. they were driven about a mile to a lupine
patch having an abundance of pods and seed. It proved to be an
unfavorable time for such an experiment, for it was very hot and the
sheep would not feed until about 7 p.m. They grazed until about

8p.m. and were kept on the move all this time. They were driven
| back to the corrals. The time of grazing was so short that it was
assumed the experiment was a failure, and the sheep were not ob-
served during the night. The next morning sheep No. 241, which

had been observed as one of those eating the most lupine, was found
down, and a little later it died, the symptoms and autopsy indicat-
ing that it was, without doubt, a lupine victim. The outcome of this
experiment was considered to be a confirmation of the general ex-

_ planation of range poisoning as given before.

TREATMENT OF RANGE ANIMALS TO AVOID POISONING.

From what has been said of the conditions under which range poi-
soning occurs, it is evident that much of the loss can be prevented by
proper management of the bands.

Sheep should never be taken from the cars to a pasture having

| much lupine. It is cheaper to buy hay. After long drives with

insufficient forage avoid grazing grounds which are covered with
lupine. If it is necessary to drive sheep over lupine patches, do not
hurry them, but allow them to spréad out and drift across. Special
care should be taken in the fall, when the grass may be covered by a
fall of snow. In the treatment of the flocks remember the general

1 fact, which is applicable in regard to all poisonous plants, that well-

fed sheep are not likely to eat injurious plants. Conditions under
which sheep get ravenously hungry should, if possible, be avoided.
Herders should recognize the fact that the pods and seeds are espe-

cially poisonous and that, consequently, poisoning is more likely to

49 BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.

occur at the time when the plants are in fruit and in seasons when
the fruit is most abundant. In very dry seasons frequently little
fruit is matured, while a wet season, especially a wet spring, may —
cause a heavy production of fruit.

SUMMARY.

(1) Lupines have been cultivated and used from the time of the
ancient Greeks and Romans, but their poisonous properties have been
recognized only in very modern times. Heavy losses of domestic
animals were reported in northern Germany in 1872 and the suc-
ceeding years. ;

(2) While chemists have shown the presence of poisonous alkaloids
in the lupines, the losses in northern Germany have been considered
by investigators as due not to the alkaloids but to a hypothetical |
substance known as ictrogen. |

(3) An investigation by Dr. Sollmann showed the presence of |
alkaloids in American lupines, and pointed to the probability that —
most, if not all, the poisoning of live stock in America was due to |
these alkaloids and not to ictrogen.

(4) Extended field work has verified the conclusions of Sollmann
and has shown that all aerial parts of the lupines examined are |
poisonous, the seeds being the most toxic, then in order the pods and ~
leaves. This has been confirmed by preliminary experiments with |
extracts upon mice.

(5) The toxic substance is excreted by the kidneys; the intoxica- .
tion is not cumulative, and animals may eat comparatively large »
quantities with no ill results, if the toxic limit is not reached at any
one time. Inasmuch as the toxic and lethal limits are nearly the |
same, the prognosis for poisoned animals is not favorable.

(6) There is no form of remedial treatment that can be used ad-
vantageously for range animals. Poisoning in most cases can be
avoided, even where the plant is abundant, by careful handling of —
the flocks, especial care being taken to see that hungry sheep are
not grazed on fields where there is much lupine.

LITERATURE CITED.

ARNOLD, CARL, and LEMKE, CARL.
1881. Beitrag zur Klarstellung der Ursache der Lupinose. Jn Deut. Ztschr.
Thiermed., Bd. 7, Heft 4, p. 251-265.
and SCHNEIDEMUHL, GEORG.
1883. Vierter Beitrag zur Klarstellung der Ursache und des Wesens der Lupinose.
In Jahresber. K. Thierarzneischule, Hannover, Ber. 15, 1882/83, p.
108-116.

BAUMERT, GEORG.
1886. Ueber den “‘ Bitterstoff,’’? das Ictrogen und Tpetcca der Lupinen. In
Arch. Pharm., Bd. 224, Heft 2, p. 49-59.

BELLINI, RANIERI.
1876. Dell’ avvelenamento prodotto dai semi del lupino. Jn Riv. Sper. Fren. e
Med. Lég., ann. 2, fase. 5/6, p. 581-627.

BERGH, G. F.
1904. Ueber die Alkaloide der perennierenden Lupine. Jn Arch. Pharm., Bd.
242, Heit 6, p. 416-440, 1 pl. :
BEYER, AD.
1871. Ueber einige Bestandtheile des gelben Lupinensamen. Jn Landw. Vers.
Stat., Bd. 14, p. 161-176.
CALLSEN, JURGEN.
1899. Ueber die Alkaloide der Samen von Lupinus angustifolius und von Lupinus
perennis var. polyphyllus. Jn Arch. Pharm., Bd. 237, Heft 8, p. 566-595.

CASssoLa.
1834. Note sur lalupinine. (Abstract by Julia-Fontenelle, with note.) Jn Jour.
Chim. Méd. Pharm. et Tox., t. 10, p. 688-689.
For German translation see Ann. Pharm. [Liebig], Bd. 13, Heft 3, p.
308-309, 1835.
CHESNUT, V. K.
1899. Preliminary catalogue of plants poisonous to stock. In U.S. Dept. Agr.,
- Bur. Anim. Indus., 15th Ann. Rpt., 1898, p. 387-420, fig. 38-69.
and Wicox, E. V.
1901. Stock-poisoning plants of Montana. A preliminary report. U.S. Dept.
Agr., Div. Bot. Bul. 26, 150 p., 36 pl.
CorneEvin, C. E.
1893. Des Plantes Vénéneuses et des Empoisonnements qu’ elles Detérminent.
524 p., illus. Paris.

DAMMANN, CARL.
1902. Die Gesundheitspflege der landwirtschaftlichen Haussiugetiere. Aufl. 3,
873 p., 74 fig., 20 col. pl. Berlin.
Davis, L. 8. .
1897. Die Lupanine der weissen Lupine. Jn Arch. Pharm., Bd. 235, Heft 3, p.
199-217.
Gemma, A. M.
1882. Studi Sperimentali e clinici sull’ azione della lupinina amorfaimpura. In
Gaz. Med. Ital. Lomb., v. 42 (s. 8, t. 4), no. 14, p. 147-149.

43

44 BULLETIN 405, U. S. DEPARTMENT OF AGRICULTURE.

Hutyra, FERENCZ, and MAREK, JOZSEF. 4
1913. Special Pathology and Therapeutics of the Diseases of Domestic Animals, |
Authorized Amer. ed. from the 3d rev. German ed., J. R. Mohler and
Adolph Eichhorn, editors, v. 2, 163 illus., 5 pl. Chicago.
Kuan, JULIUS.
1880. Die Schmarotzerpilze der Lupinenpflanze und die Bekimpfung der Lupin-
_ enkrankheit der Schafe. Jn Ber. Physiol. Lab. u. Vers. Anst. Landw.

Inst. Halle, Heft 2, p. 115-128.
LANDERER.

1852. Ueber den Bitterstoff aus Lupinus. Jn Neues Repert. Pharm., Bd. 1,
~ Heft 10, p. 445-447.
LIEBSCHER, G.
1880. Beitrag zur Klarlegung der Frage nach den Ursachen der Lupinenkrankheit
der Schafe. Jn Ber. Physiol. Lab. u. Vers. Anst. Landw. Inst. Halle, Heft
2, p. 53-114.

LOWENTHAL, JULIUS.
1888. Ueber die physiologischen und toxicologischen Eigenschaften der Lupinen-
Alkaloide. 26 p., Kénigsberg. Inaugural-Dissertation.
MUENK, GUSTAV.
1914. Beitrage zur Kenntnis der Bestandteile und Wirkungen der Lupinensamen,
In Landw. Vers. Stat., Bd. 85, Heit 6, p. 293-416.

Puintus SEcUNDUS, CatUS. ; |
1856. The Natural History of Pliny. Tr... by... John Bostock and H. T. |
Riley, v.4. London.
Ramonnt, C. :
1891. Sur les principes actifs et toxiques du lupin. Jn Arch. Ital. Biol., t. 15,
fasc. 2, p. 246-253. ‘4
Ro.LorFr, FRIEDRICH. =.
1883. Ueber die Lupinose. Jn Arch. Wiss. u. Prakt. Thierheilk., Bd. 9, Heft ©
4, p. 1-54.
Scumipt, ERNST.
1897. Ueber die Alkaloide der Lupinensamen. Jn Arch. Pharm., Bd. 235, Heit 3,

p. 192-198.
1904. Ueber die Lupinenalkaloide. Jn Arch, Pharm., Bd. 242, Heft 6, p. 409-415
Suave, H. B.

1903. Some conditions of stock poisoning in Idaho. Idaho Agr. Exp. Sta. Bul.
37, p. 159-190, illus.
Witcox, E. V.
1899. Lupines as plants poisonous to stock. Jn Mont. Agr. Exp. Sta. Bul. 22, p. |
37-45. Literature, p. 43-45.
WILusTATTER, RIcHARD, and FOURNEAU, ERNEST.
1902. Zur Kenntnis des Lupinins. Jn Arch. Pharm., Bd. 240, Heft 4, p. 335-344,
Zien, F. A.
1879. Massenerkrankung von Schafen durch den Genuss beiallener Lupinen. In
Vortr. Thierirzte, S. 2, Heft 7, 29 p. (p. 251-277), 6 fig. ;

PUBLICATIONS OF THE U. S. DEPARTMENT OF AGRICULTURE RELATING
TO POISONOUS PLANTS.

AVAILABLE FOR FREE DISTRIBUTION BY THE DEPARTMENT.

Zygadenus, or Death Camas. (Department Bulletin 125.) .
Larkspur Poisoning of Live Stock. (Department Bulletin 365.)
_ Larkspur, or ‘‘ Poison Weed.’’ (Farmers’ Bulletin 531.)
Prevention of Losses of Live Stock from Plant Poisoning. (Farmers’ Bulletin 720.)

FOR SALE BY THE SUPERINTENDENT OF DOCUMENTS, GOVERNMENT PRINTING
OFFICE, WASHINGTON, D. C.

-Cicuta, or Water Hemlock. (Department Bulletin 69.) Price, 10 cents.
The Poisonous Action of Johnson Grass. (Bureau of Plant Industry Bulletin 90,
Pt. IV.) Price, 5 cents.

The Larkspurs as Poisonous Plants. (Bureau of Plant Industry Bulletin 111, Pt. I.)
=) rice, 5 cents.
The Supposed Relationship of White Snakeroot to Milksickness, or ‘‘Trembles.”’

(Bureau of Plant Industry Bulletin 121, Pt. I.) Price, 5 cents.

Mountain Laurel, a Poisonous Plant. (Bureau of Plant Industry Bulletin 121, Pt. II.)
Price, 5 cents.

Results of Loco-Weed Investigations in the Field. Laboratory Work on Loco-Weed
Investigations. (Bureau of Plant Industry Bulletin 121, Pt. III.) Price, 5 cents.

Barium, a Cause of the Loco-Weed Disease. (Bureau of Plant Industry Bulletin 129.)
Price, 10 cents.

The Relation of Barium to the Loco-Weed Disease. (Bureau of Plant Industry Bul-
letin 246.) Price, 10 cents.

Menziesia, a New Stock-Poisoning Plant of the Northwestern States. (Bureau of
Plant Industry Circular.) Price, 5 cents.

The Loco-Weed Disease of the Plains. (Bureau of Animal Industry Bulletin 112.)
Price, 35 cents.

The Stock-Poisoning Plants of Montana. (Division of Botany Bulletin 26.) Price,
25 cents.

The Loco-Weed Disease. (Farmers’ Bulletin 53k.) Price, 5 cents.

Stock Poisoning Due to Scarcity of Food. (Farmers’ Bulletin 536.) Price, 5 cents.

45

.

WASHINGTON : GOVERNMENT PRINTING OFFICE : 1916

ADDITIONAL COPIES
OF THIS PUBLICATION MAY BE PROCURED FROM
THE SUPERINTENDENT OF DOCUMENTS
GOVERNMENT PRINTING OFFICE
WASHINGTON, D. C.
AT

10 CENTS PER COPY
AY

LIBRARY

|
|
43

WIT
10227898