Lathyrism in the Rat Geiger 1932 427

7/29/2019 Lathyrism in the Rat Geiger 1932 427

1/16

LATHYRISM IN THE RATByBEATRICE. GEIGER,HARRYSTEENBOCK,ANDHELENT. PARSONS*(From the Departments of Agricultural Chem istry and Home Economics,U niversity o f W isc on sin , M ad iso n)

Received for Publi ca t ionOc tober 6 . 1932

E^THYRISM , which has been known since the time of Hippocrates,has been very common in the past in India and northern Africa, andsm all local outbreaks have occurred frequently in Italy, France, and elsewhere in southern Europe (1). For a long time it has been known that thedisease is caused by the eating of lathyrus peas of three species, Lathyrussativus, Lathyrus ccera,and Lathyrus clymenum. After a poor crop ofwheat, barley, and other cereals, the poorer people in India and som e partsof northern Africa are forced to eat the lathyrus peas as a large part oftheir diet, and lathyrism then becomes prevalent. The use of the peas asone-third to one-half of the diet for two or three months is consideredenough to cause the disease (2). However, not all persons eating such adiet are affected, and in fam ilies only certain m em bers are attacked. W henthe peas are eaten as a smaller proportion of the diet, they are said to beh arm le ss (1 ).The outstanding symptom of lathyrism in man is a spasticity and rigidity of the leg muscles which results in a jerky gait w ith flexed knees, or aninability to w alk. The condition develops very rapidly and is often precipitated by exposure to cold, w et, and fatigue. D eath seldom results, but thereis usually a permanent lameness (1). The symptoms have suggested tovarious workers the possibility that they may be due to degenerativechanges in the spinal cord. In fact Mellanby (3) has gone so far as to suggest that vitam in A may be able to prevent this degeneration. However,he has not reported any experiments to test this idea so far as we areaware.Different anim als are said to vary greatly in their susceptibility to thepoisonous action of the lathyrus peas. In Great Britain peas of this typeare used as food for farm animals. When mixed in the ration in not too* Thi s p ap er w as s ubmi tt ed b y B ea tr ic e J . G ei ge r in p ar tia l f ul fi llm en t o f th e re qu ir em en ts f ort he Ph.D . d eg re e, 1 93 2. P ub li sh ed w it h t he p erm is sio n o f t he D ir ec to r o f th e W i sc on si n Agr ic ult ur al Expe rimen t S ta ti on , Mad is on .

42 7


2/16

428 LATH YRIS M IN THE RAT Vol.6, No. 5

large proportion they are said to give satisfactory results, although occasionally bad effects have been reported (1). Of the farm anim als, horseshave been found to be peculiarly susceptible. Stockman (1) stated that20 per cent of lathyrus peas in the diet is harmful to horses, while pigs andcattle can tolerate higher levels. Feeding experim ents have been carriedout using m onkeys, rabbits, guinea pigs, rats, pigeons, and ducks. A ccording to Stockman (4) m onkeys have spasmodic attacks of a dramatic kindoccurring at irregular intervals, in addition to the paralysis. He reportedthat death w as due to failure of the respiratory m uscles. In his experim entson rabbits, in which he used seven animals, he observed in some cases avery transient and slight spasmodic condition of the legs, but most of hisanim als grew normally. He assumed the presence of a toxin and inferredthat rabbits are susceptible to the toxin, but that in ordinary feeding experiments they get too little poison to produce symptoms. In four out offive guinea pigs he noted tremors, spasticity, and weakness of the legsand reported that all died in 8 to 35 days. He stated that "the lethal actionis not a specific lathyrus action, but seems to be due to the fact that theproteins of lathyrus peas cannot be assim ilated and utilized by guineapigs." McCarrison (5) reported that he was unable to cause lathyrism inrats even when feeding lathyrus peas as the only food. Zagam i (6) alsofailed to secure sym ptom s characteristic of lathyrism in rats. H e fed a dietmade up entirely of lathyrus peas for 150 days. He did report a decreasein the rate of gain, retardation in sexual development, and a decrease incalcification of the skeleton, which can be readily understood, inasm uchas all seeds w hen fed by them selves are seriously deficient in various nutritive elem ents. Acton is said to have produced paralysis in ducks by feedingthem lathyrus peas (1). Pigeons have been reported insusceptible to theto xin (4 ).The discrepancies in the results of different investigators are difficult ofinterpretation. Undoubtedly many of them were due to the use of poorbasal diets which frequently were deficient in m ineral elem ents, vitam inD , and good protein, and probably were otherwise poorly constituted.M ellanby (3), for instance, believes that vitam in A furnished variable protection depending on the am ount present. Em phasis should, however, alsobe placed on the fact that the investigators frequently used different species and varieties of lathyrus peas and no mention was made of the conditions un der w hich they w ere grow n.There is a difference of opinion among experimenters as to the natureof the toxic substance in the lathyrus peas. Stockman (7) asserted that


3/16

September,1933 GEIGER, STEENBOCK, AND PARSONS 429

the tox ic principle w as an alk aloid w hich he obtained in v ery sm all am ountfrom the peas and w ith w hich he could produce paraly sis in frogs, m onk ey s, and m ice. How ever, A cton and Chopra insisted that an am ine andnot an alk aloid w as the tox ic substance (cited from [l]). T hese inv estigators reported paraly sis in the hind legs of m onk ey s, guinea pigs, rats, andm ice w ith the pure hy drochloride of the am ine obtained from the peas.In contrast to both of these v iew s, A nderson, How ard, and S im onsen (8)are of the opinion that the poisonous ef fect w as due to other seeds, especially V icia sativ a v ariety A u gustif olia, f ound m ix ed w ith the lathy ruspeas. T hey observ ed no tox ic ef fects of any k ind w hen botanically pureLa thyrus sa livus seeds were fed to anima ls, but sta ted tha t when seeds ofVicia sa tiva were fed to monkeys, nervous and muscula r symptoms resulted, and w hen fed to duck s they caused death. V icia sativ a w as reported as containing bases show ing alk aloidal properties of w hich theyisolated three, v icine, div icine, and a cy anogenetic glucoside, v icianin, atleast one of w hich, div icine, produced characteristic sy mptom s w hen injected into guinea pigs. A still dif ferent theory of the cause of lathy rismw as advanced by Y oung (2) af ter he m ade a f ield study of the disease inIndia. He suggested that the disease w as due prim arily to a def iciency ofv itam in A .From the preceding it is ev ident not only that there is considerabledif ference of opinion regarding the etiology of lathy rism , but also that theproperties of dif ferent species are v ery d if ferent and hard ly specif ic.In the ex perim ents detailed in the follow ing report the ef fect of feedinglathy rus peas to rats w as determ ined. A n attem pt w as m ade to ascertainw hether the abnorm al condition produced by the peas w as am eliorated orprev ented by the feeding of cod liv er oil or y east as v itam in supplem ents.T he species of lathy rus peas w hich w as used in our experim ents w asLa thyrus odora tus, the sweet pea used for ornamenta l purposes in thiscountry.

EXPERIMENTALT he com position of the diets is show n in T able I.1 T he casein w as a comm ercial product f reed from v itam in A by heating it at 90 . for tw o w eek s.T he dex trin in reality represented cook ed starch. It w as prepared by m ixing enough w ater w ith cornstarch to m ake lum ps, autoclav ing it for 21 W e w ish to thank John B odger and S ons of El M onte, C alifornia, f or the sw eet peas, E. R .S quibb and S ons f or the cod liv er oil, and N ational O il Products C om pany f or the cod liv er oilconcentrate used in these experimen ts .


4/16

4 30 LATH Y RIS M IN THE RAT Vol. 6, No. 5

Bao

0

O Oio ^H

1 1

3

O OIO r-1

o oIO -! o csio

0 0IO '-'"*

l l S M l l l

^

c nQUO

e i8 ~St,1 1 lII CaCNaCl

'S .M!!- in iB|IJ*"? l

s -s.S g fa a ja 3IJl*s "n^ tf10a s s 11^-

'S9 IT!So 320


5/16


hours at 15 Ibs. pressure, dry ing it, and then grinding. T he cod liv er oilconcentrate w as incorporated in the ration by dissolv ing it in ether, pouring the ether solution on part of the dex trin and then evaporating of f thesolv ent in a current of air. T his w as then m ix ed w ith the other ingredientsof the ration. T he y east w as obtained from the N orthw estern Y east Company and had been prev iously tested in this laboratory for its v itam in Bcontent. T he irradiated y east w as y east w hich had been irradiated for 2hours w ith an A lpine S un Lam p at a distance of 18 inches w ith stirring.T he rations w ere so outlined as to be adequate in all respects, ex ceptthat in som e, v itam in A w as absent aside from such am ounts as w ere contained in the sw eet peas them selv es. For control purposes to dem onstrate the general adequacy of the supplem ents, garden peas w ere substituted for the sw eet peas at the higher lev els and 2 per cent of cod liv eroil w as included. W ith these in the diet approx im ately norm al grow th w asobtained and the anim als m aintained them selv es in ex cellent condition f or14 w eek s. A t this tim e the trials w ere term inated. T he rations w ere fed adlibitum, the food consumption being r ecorded dur ing the fir st 4 or 5 weeksof the ex perim ents. M ale albino and piebald rats w eighing betw een 50 and60 gram s w hen 3 to 4 w eek s old w ere used in lots of 4. Each rat w as k eptin a separate cage w ith a raised screen bottom . Distilled w ater w as giv enin addition to the ration. T he anim als w ere w eighed once a w eek .Contrary to the f indings of M cCarrison (5) and Z agam i (6) w e hav e succeeded in inducing lathy rism in rats. V ery little grow th occurred on thediets w hich contained 80 per cent of sw eet peas and the rats liv ed for only4 to 6 w eek s (L ot 4). A s the percentage of sw eet peas in the diet w as decreased, greater grow th resulted. Of the 40 anim als receiv ing 50 per centof sw eet peas from the tim e of w eaning, 3 grew to a w eight of 160 gram s,during 3-1/2 m onths on the ration; the rest w eighed less than this. One ofthe 8 anim als on the 25 per cent lev el attained a w eight of 215 gram s. However, w hen the sw eet peas w ere fed as 12.5 per cent of the ration or less,practically norm al grow th w as secured for 20 w eek s in a group of 8 rats.T he grow th of the anim als is sum m ariz ed in T able II.Depression of grow th w as only one ef f ect of the diets containing sw eetpeas. T he f irst sign of abnorm ality appeared af ter about 4 w eek s on dietscontaining 50 per cent of sw eet peas and af ter 6 w eek s on those containing25 per cent. A h1 the anim als dev eloped a m arked spinal curv ature. T helong bones w ere v ariously deform ed, and the rat f requently w alk ed w ith asham bling gait. T he abnorm alities becam e progressiv ely w orse and apparently the anim als w ere in discom fort, as they becam e very inactiv e.


6/16

\ -

4 32 LATHY RI S M IN THE RAT Vol. 6, No. 5

TABLEnGROWTHOPRATSON SWEETPEA DIETS(Average weights)

LotNo.143262739505557Characterfration50%

gardenpeas+2%C.L.O.80%

sweeteas50%

sweeteas50%

sweeteas50%

sweet peas+2%.L.O.50%sweet peas+2%.L.O.50%sweet peas+2%.L.O.50%sweet peas+2%C.L.O.+0.1%.O.C.t50%

sweet peas+10% yeastWeightInitialgms.

565254555555585457at

4weeksgms.

1137510411110385107101121at

6weeksgms.

136112123125114132113132at

10weeksgms.

168123118143143143127at

15weeksgms.RemarksControl

group. Approximatelynormal growth. In excellent condi ti on when etherized af ter 13weeks.V ery

litt le growth. Digestive disturbance (flatulence, diarrhea).Respiratory difficulty after 3weeks. H ind legs of 2 rats paralyzed after 5 weeks. All died during 5th or 6theek.Slowgrowth for 4 to 6 weeks.H ind legs lame after 4 weeks,those of 2 rats in L ot 3 almostcompletely paralyzed. Lordosis inthoracic region. Sternum misshapen in 2 rats. Some digestivedisturbance. 3 died between 7thand 12th weeks, otherstherized.Someimprovement in growth withaddition of C.L.O. Lameness andlordosis af ter 4 or 5 weeks. Hindlegs partially or completely paralyzed. All rats in Lot 27 developedhernias. Ventral or lateral spinalcurvature inall. 4 died between 5thand 12th weeks, otherstherized.No

better growth than Lots 27, 39and 50. Lameness and lordosisafter 5 weeks. One rat developeda hernia. Etherized after 7eeks.Nobetter growth than Lots 3 and26. Symptoms of lathyrism after5 weeks. All died between 6th and15th weeks.

t C.O.C.=Cod liver concentrate.


7/16

September ,933 G EIG ER, STEENBOCK, AND PARSONS 433TABLEn (Continued)

LotNo.SI528182282973747576Characterfration25%

sweeteas25%

sweeteas+2%C.L.O.12.5%sweetpeas5%

sweeteas50%

sweeteas(cooked)50%

sweeteas(cooked)+2%C.L.O.50%

sweeteas50%

sweet peas+2%C.L.O.25%

sweeteas25%

sweet peas+2% C.L.O.WeightInitialgms.545453575255359343383342at

4weeksgms.918913911792102345340363337atweeksgms.118112185182100117348352372346at0weeksgms.158159249208100135347351366347at15weeksgms168176277242350346377349RemarksBettergrow th than w ith0%.Growthfairly g oo d for 1 2eekswithno C.L.O. in die t. Symptomsof lathyrism after 6 weeks . Etheri zed af te r 18eeks.Slightlybe tte r g rowth thanot51.S ymptoms o f l athy ri sm i nratsafte r 6 w eeks, in o ne afterweeks.Etherized af ter 18eeks.Normalg rowth w i th no sourc efvitaminA exc ep t swee t peas .osymptomso f l athyri sm af te r0TaArc.Vetnt ! *ol A i . Ti .weeks

to supplement v itamin Bnsweetpeas s ince appe ti te was dec lining . Etherized a fter 20eeks.Slightenlargement o fostochon-draljunctions onutopsy.Samere sults as w ithncookedsweetpeas. 5 die d betw ee nthand12th weeks , o therstherizedafter12eeks.Weightmaintamed. Lameness appeared in al l ra ts . Etherizedfter15weeks.Slightincrease in w eight. N olamenes s deve loped. E theri zedaf ter 15eeks.Weightmaintained. N o symptoms of lathyrism. Etherizedfter15weeks.Slightincrease in w eight. Nosymptom s o f lathy rism . Ethe rized af ter 15weeks .


8/16

4 34 L AT H Y RI S M IN THE RAT Val. 6, No. 5TABLEn (Continued)

Lo tNo.78778379117121Characteroration50%

sweeteas(novitamin.)#2965+50%sweet

peas#2965+50%sweet

peas+2%C.L.O.Control

DietWater

extractofsweeteas50%water extractedweetpeasWeightInitial515455535454at

4weeks90918214613573at7weeks1118893207129116at10weeks12679102256134203at15weeksRemarksSlightlypoorer growth thanots3and 26 which receivedrradiatedyeast.A ll di ed be tw ee n 3 rdnd14thweeks.Lathyrismaf te r 3 weeks . S ixiedothers

etherized.Normal

g rowth . E theri zed af te r11eeks.Go odgrow th for 4 w eeks,hendecline.Lathyrism after 4eeks.AUdied betw een 7th and2thweeks.Goodgro wth. N o symptomsflathyrismaf te r 11 weeks. Expe riment continuing.

In a few cases the hind leg s w ere completely paralyzed. In 2 cases the frontlegs were also involved, the feet turning out at the wrists. In many of theanimals the sternum was deformed. For a typical illustration of suchanimals see Plate 1.A lthoughhe rnia in the rat is a v ery unusual phenomenon, prac tic all ynever occurring in the stock co lony and to the best of our know ledg e neverhaving been reported in relation to dietary abnormality in the rat, it occurred in 10 cases, or 25 per cent of the young animals receiving the 50per cent level of sweet peas. Some of the hernias were permanent whileo thers appeared and disappeared at interv als . The co ntents o f the herniasvaried, sometimes consisting of the cecum and part of the intestines o nly,w hile in others a kidney, a testis, and even the stomach and spleen protruded in addition.The severity of the skeletal changes became evident on autopsy. Thespinal column in every case showed extreme curvature in the thoracic


9/16

September , 1933 GEIGER, STEENBOCK, AND PARSONS 435

regi on. In mos t o f the animals the curvature was only ventral although itsome times was also late ral. Invariably the v entral curvature involv ed

PLATE1. As ide v iew o f o ne rat from Lo t 3 s howing the s pi nal c urv ature c harac te ri stic o fan imal s on d ie ts c on taining 50and 25per c en t o f swee tpeas .the lower seven thorac ic vertebrae . An outs tanding feature was the shortradius whi ch the curvature as sumed even in the larger animals . This did

199

;ut>- ILLS'

PLATE2.Photographs of vertebral columns showing curvatures in the thoracic region. Thesepho to graphs w ere take n w ith the v entral s ide uppe rmos t and the c erv ic al e nd to the le ft. R at4 93anormal an imal f rom the s to ck col ony. Rats 1 97 , 1 99 , 2 00 f rom Lot 30 re ce iv ed a d ie t c ontaini ng 5 0 pe r c ent o f sw ee t pe as and 2% o f c od li ve r o il . R ats 2 25 , 2 2 6, 2 28 f rom Lot 5 7 re ce iv eda die t c ontai ning 5 0 p er c ent o f sw ee t pe as and 1 0 pe r c ent o f y eas t. R at 3 12 f rom Lo t 7 8 re ce iv ed50 per c en t o f swe et peas .


10/16

436 L AT H YRI S M IN THE RAT Vol.6, No. 5

not exceed 3/8 of an inch (Plate 2). In most cases there was also a dorsalcurvature in the lumbar region. In some rats the tail made an acute anglew ith the body.Although the spinal curvature dom inated the picture the thoraciccavity was otherwise affected. In the first place the sternum was frequently deform ed. Som etim es it assum ed a dorsal-ventral curvature, butin other cases it was also deform ed laterally . It was noted that in additionthe ribs were frequently m isshapen. Som etim es the costochondral june-

"

493'

... 197*

PLATE3 . Pho to gr ap hs s how ing ma lf ormat io n o f r ib s a nd s te rn ums . Ra t 4 93 Anormal a nim al from th e sto ck c olo ny . R ats 1 07 a nd 19 7 FromL ots 2 7 an d 5 0 re sp ec tiv ely re ce iv ed a d ietco nta in in g 5 0 p er ce nt o f s w ee t p ea s a nd 2 p er ce nt o f co d liv er oil. N ote a ngu latio n a t the c os toc hond ra l j un ct io ns a nd d ef ormat io n o f t he s te rn um .tions were merely enlarged, but in other cases angulation was also pronounced. The photographs shown in Plate 3 may be taken as typical examples. It was evident that these changes could not have been caused byrickets, since all the rations contained 4 per cent of a complete salt m ixture as well as vitam in D in an am ount previously shown in this laboratoryto b e amp le fo r p ro te ctio n.The femora and numeri were badly deformed. The upper ends of thefem ora were broadened and rough areas were present on either side at thepoints of attachment of the muscles. The humeri were also thickened and


11/16

y i ..-vje;-

September,1933 GEIGER, STEENBOCK, AND PARSONS 437show ed sim ilar roughened areas (P late 4). Taking an average of 30 anim alsand comparing them with a sim ilar number which had sim ilar weightsrecorded by Donaldson (9), the weights of femora and humeri were foundto be 33 and 26 per cent less, respectively.In view of^thc deform ities both in the vertebral column and in the longbones it was believed possible that changes in m ineralization of the boneshad occurred. However, that this was not extreme was evident from thefact that the bones were observed to be neither brittle nor flexible. D eter-

V ^^WMV1


12/16

438 LATH Y RIS M IN THE RAT Vol. 6, No. 5

table that the ash content of the bones of the adults falls within the normal limits. In the young both the amount of ash and percentage of ashwere decreased. TABLEIIIPERCENTAGEKASHIN THEBONESOFRATSONSWEETPEA DIETS

Lot26,2827,

29 ,95557515273747576StockStockNo.

offemora36211322221514Sweetpeadditions50%50%+2%

C.L.O.50%+2%C.L..+0.1%C.O.C.t50%+

10%east25%25%+2%C.L.O.50%50%+2%C.L.O.25%25%+2%C.L.O.StockrationnlyStockration onlyAverage

agedays11310077114161156274281247257101201Averageweightgms.116136116xtractedfemurgms.0.3170.3370.2870.2990.5300.4710.6510fshgms.0.1680.1770.1490.1730.3360.2930.4000

t C.O.C. = Codl iver o il concentra te .Inasmuch as the blood phosphorus and calcium relations have served

as valuable indices of the state of bone metabolism, such data on ouranimals were obtained. The Fiske-Subbarow method (10) was used fordetermination of inorganic phosphorus. The serum calcium w as determined on the protein-free filtrate obtained after treating the serum w ithfour times its volume of 10 per cent trichloracetic acid. Aliquots corresponding to 1 cc. of serum w ere used. The hydrogen ion concentrationw as adjusted in the manner used by McCrudden (11) for organic materials. The calcium was precipitated with ammonium oxalate and aftertreatment with normal H2S04 was titrated w ith N/100 KMnO4, using amicro -burette. The v alues o btained are tabulated in Table IV . Inspectio nof the collected data on the adults reveals that the serum calcium andino rg anic pho spho rus w ere w ithin the no rmal limits . U nfo rtunately blo odanalyses on young rats w ere no t made.One of the outstanding changes in the bones was their congested appearance. D istinct hemorrhages w ere also no ted macroscopically in the


13/16

September,1933 GEIGER, STEENBOCK, AND PARSONS 43 9mu scle an d c onn ectiv e tissu e imme diate ly c on tig uo us to the ro ug hen edareas on som e of the fem ora and num eri. T he appearance of the bonesth em s elv es sug ge sted a p ossib le stim u latio n o f h emato po ietic ac tiv ity .In div id ual d ete rm i natio ns o f h emog lo bin b y th e N ewcomer me th od , u sin gblood obtained f rom the tails of 23 of the anim als, gav e results rangingf rom 12.2 to 15.9. T hree-f ourths of the v alues fell abov e 14, w hich in-

TABLEIVMILLIGRA MSER CENT OPCA LCIUMA NDPHOSPHORUSN THEB LOODOFRA TSO N S W EET PEA D IET S

Lot737451757652No.ofrats444344Sweet peadditions50%50%+2%C.L.O.25%25%25%+2%C.L.O.25%+2%

C.L.O.Serum

Camg.%10.411.210.09.910.010.2Serum

Pmg.%6.97.09.16.96.69.6

dicates a tendency tow ard a high lev el. T he ery throcy te count of thesesame an ima ls e stima te d in a L e v y countin g chambe r w i th sin gle N eubau erruling av eraged 9,900,000 w ith a range of 7,800,000 to 12,000,000. AUbut 3 counts w ere abov e 9 m illion and 10 w ere abov e 10 m illion, thusf alling in the upper part of the norm al z one or slightly abov e. A v eragedvalues of three to four anim als in each group are show n in T able V .Ex am ination of the spleens show ed them to be som ewhat sm aller in pro-

TABLEVHEMOGLOB INALUESA NDERY THROCY TEOUNTSOFRA TSONSW EETPEA D IET SLot737451757652No.frats444344Sweet peadditions50%50%+2%

C.L.O.25%25%25%+2%C.L.O.25%+2%

C.L.O.Hemoglobin,

gm s. per 100cc. oflood15.4814.7514.2014.0213.5514.26Erythrocytespe rcu. m m . oflood9,863,00010,163,0008,850,00010,850,000


14/16

440 LATHY RI SM IN THE RAT Vol. 6, No. 5portion to the body weight, e spe cially in tho se animals re ce iv ing the 50per cent leve l o f sweet peas (Table VI) .

TABLEVITHE WEIGHTOF THESPLEENOF RATSON SWEETPEA DIETS

Lot732627295055577551765279No.frats421144334443Sweeteadditions50%50%50%+2%

C.L.O.50%+2%C.L.O.50%+2%C.L.O.50%+2%C.L.O.+0.1%C.O.C.f50%+

10%east25%25%25%+2%C.L.O.25%+2%C.L.O.Control

dietRange

ofbodyeightgms.294-400125-147 1471401 23-151109 -12099-13 8351-4271 30-19831 3-3spleenweightgms.0.7400.2700.3110.3670.2510.5890.3720.6240.4610bodyweightdividedbyveragespleenweight4755074713815492433436073754

t C .O.C . = Cod l iver oi l concentrate.

To suppl ement our obs ervations on young animals a f ew experimentswere carried out on o ld rats. They were put on the ration at a weight o f 3 00to 400 g rams. S ome o f the g ro ss e ffe cts obtained w ith the young animalswere s ecured with such adults . The femora and numeri became markedlyconges ted, the muscular attachments were hemorrhag ic and the cortexwas roughened. A ll the long bone s were de fo rmed. Lamene ss be came inc ident after 1 2 w eeks in the animals which rec eiv ed 5 0 per ce nt o f sw ee tpeas w ith no v itamin supplement. The ess enti al diff erences were that thespinal columns , the ribs , and s ternums were normal. Hernia als o did no tdevelop. !&&%*!''NWiTo pave the way for further s tudi es on lathyrism, but no t in direct re latio n to o ur immediate pro blem, an attempt was made to inac tiv ate theto xic princ iple in the swee t peas by cooking them fo r 2 -1 /2 hours. How

ever, no decreas e in the s everity o f the toxic e ff ec t could be no ted when thecooked peas to ge the r w ith the wate r in which they were cooked were fedat the 5 0 per ce nt le vel. Later the to xic principle was found to be waterso luble and fairly stable to heat at the bo iling po int o f wate r. In the se


15/16


experim ents the peas w ere soaked over night in distilled w ater and w erethen brought to the boiling point. A f ter standing for 30 m inutes the w aterw as poured of f . T he peas w ere then cov ered again w ith distilled w ater andthe ex traction w as repeated as before, f iv e tim es. A f inal ex traction w asm ade w ith a large am ount of w ater, this ex tract being discarded. T he othercom bined ex tracts w ere concentrated to a sm all v olum e at 60 to 65 C. bef ore an electric f an and w ere then d ried on dex trin at the sam e tem perature.W h en this w as incorporated in the ration so that each 100 gram s of rationcarried an am ount of ex tract equivalent to 100 gram s of sw eet peas, thesam e tox ic ef fects w ere observ ed as w hen 50 per cent of sw eet peas w ere f ed.Furtherm ore, the ex tracted sw eet peas f ed at the 50 per cent lev el producedno apparent tox ic ef fects af ter 11 w eek s.A n attem pt to prev ent or correct the sym ptom s of lathy rism w as m adealong the lines suggested by M ellanby (3) by v itam in additions. Codliv er oil did not protect the young anim als from the tox ic ef fect of thehigher lev els of sw eet peas. In fact w ith 0.1 per cent of cod liv er oil concentrate in addition to the 2 per cent of cod liv er oil no decrease in thesev erity of the sk eletal changes could be noted. Dried y east w as also w ithout protectiv e ef fect. How ev er, the adult anim als w ere protected to som eex tent by 2 per cent of cod liv er oil at both the 50 and 25 per cent lev els,w hich w as apparent in the condition of both the f em ora and hum eri. T hebones of the anim als receiv ing the cod liv er oil w ere sm aller, had few erex ostoses, and w ere m ore nearly of the sam e color as the controls.It is notew orthy that so m uch grow th w as possible w hen no v itam in Aw as present in the diets other than that furnished by the sw eet peas. W ithonly 5 per cent of sw eet peas in the diet norm al grow th w as secured for 20w eek s. T his indicates that the sw eet peas used by us w ere unusually richin v itam in A as com pared w ith other seeds.It is possible that the v itam in A in the sw eet peas ex erted a substantialneutraliz ing ef fect ev en w ithout the addition of cod liv er oil. It is suggested that future studies designed to determ ine the neutraliz ing ef fectof v itam in A be carried out w ith w ater ex tracts inasm uch as these can bereadily obtained free from v itam in A . How ev er, w e believ e that ev en w ithour present technic w e discov ered som e prophy lactic action w ith cod liv eroil. T he results seem ed to be clean-cut w ith our adult anim als.

SUMMARYL athy rism w as produced in both young and adult rats by feeding dietsw hich co ntained L athy ru s odo ratus, the f low ering sw eet pea.


16/16

442 LATHY RISM IN THE RAT Vol.6,No. 5

Growth of the young animals was retarded by sweet peas when theywere fed as 80 per cent, 50 per cent and 25 per cent of the diet. However,normal growth was obtained for 20 weeks with 12.5 per cent and 5 percent of sweet peas. O ther symptoms of lathyrism noted were lameness,spinal curvature, sternal curvature, enlargem en t of the costochond ral junctions, and malformation and abnormal red color of the long bones. Calcification w as interfered w ith in young anim als. The decided reddish colorof the bones harm onized with a slight polycythemia and increased hemoglobin con tent of the blood. H ern ias w ere also o bserved.The young animals received no protection against the toxicity of thesweet peas from cod liver oil or cod liver oil concentrate, or from driedyeast. However, the adult animals were protected to some extent by theinclusion of 2 per cent of cod liver oil in the ration.C ooking the sweet peas for 2-1/2 hours did not destroy their toxicity.The toxic factor was extractable from the peas by water at the boilingpoint.The sweet peas used by us proved to be a better source of vitam in Athan any other seeds previously studied, since norm al growth was securedon a diet in which 5 per cent of sweet peas was the only source of vitam inA. BIBLIOGRAPHY

1 . S to ckma n, R ., L ath yr ism. J ou r. P ha rm . Exp tl . T he ra p., 1 92 9,3 7,4 3.2 . Y ou ng , T . C . M ., A F ield S tu dy o f L a th yrism . In d. Jo ur. led .R es., 1 927 , IS , 4 53 .3 . M ella nb y, E ., A L ec tu re on th e R elatio n o f D ie t to H ealth a nd D ise as e. S om e R ece nt In ve stig at io ns . B r it . Med. J ou r. , 1 930, 1, 67 7.4 . S to ckm an , R ., L at hy ri sm i n Ma n. E di nb ur gh Me d. J ou r., 1 91 7,1 9, 2 97 .5 . McCa rr is on , R ., S tu die s o n L ath yr ism. I nd . J ou r. Me d. R es ., 1 92 7, I S, 7 97 .6 . Z ag am i, V ., S ug li e ff et ti d el l' a lim en ta zi on e e sc lu si va c on s em i d i " La th yr us S at iv us L ." n elr at ti a lb in i. A tt i d ell a R ea le A cc ad em ia N az io na le d ei L in ce i. 6 th S er ie s. R en di co nti , 1 93 1, 9 ,218.7 . S to ckman , R ., La th yr ism . Ed in bu rg h Med . J ou r. , 1 917, 19 ,2 77 .8. A nderson, L . A . P ., H ow ard, A . and S im onsen, J. L ., S tudies on L athyrism . Ind. Jour. M ed.Res., 1924-25,12,613.9 . D on ald so n, H . H ., T he R at. M em oirs o f t he W ista r In stitu te o f A na to my . N o. 6,1 92 4.10. F iske, C . H ., and S ubbarow , Y ., C olorim etrie D eterm ination of P hosphorus. Jour. B iol.Chem. , 1925,66,375.1 1. M cC rud de n, F . H ., T he D eterm in ation o f C alcium in the P res en ce o f M ag nesiu m an d P ho spha te s. Th e De te rm ina ti on o f Ca lc ium in Ur in e. J ou r. B io l. Ch em . , 1 9 11- 12 ,1 0, 18 7.

T he au tho rs w ish to ex pres s th eir ap prec ia tio n to Ja me s T . L ow e, C arl B auma nn an d E vely nC . V an Don k f or a ss is ta nc e in t he a na ly tic al w or k.

Documents

Lathyrism in the Rat Geiger 1932 427