`
`
`
`TREASURY DEPARTMENT
`
`
`
`
`
`PUBLKJHEALTH
`
`REPORTS
`
`ISSUED WEEKLY
`BY THE
`
`UNITED STATES PUBLIC HEALTH SERVICE
`
`
`CONTAINING INFORMATION ON THE CURRENT
`PREVALENCE OF DISEASE, THE PREVENTION
`AND CONTROL OF DISEASE, AND
`RELATED SUBJECTS
`
`VOLUME 43—PART 1
`NUMBERS 1—26
`
`J ANUARY—J UNE, 1928
`
`
`
`UNITED STATES
`GOVERNMENT PRINTING OFFICE
`WASHINGTON
`1928
`
`
`
`
`
`Elysium Health Exhibit 1013
`Page 1 of 22
`
`

`

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`
`ADDITIONAL COPIES
`OF THIS PUBLICATION MAY BE PROCURED FROM
`THE SUPEIUNTENDENT OF DOCUMENTS
`1T.S.!}O\'EI{NA1ENT PRINTING OFFICE
`WASHINGTON, D. C.
`AT
`5 CENTS PER COPY
`
`-,n:1r<r :151
`
`\V:
`
`Elysium Health Exhibit 1013
`Page 2 of 22
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`

`

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`
`PUBLIC HEALTH REPORT§
`
`
`T’OL. 43
`MARCH 23, 1928
`N0. 12
`
`
`SMALLPOX IN THE UNITED STgTES
`PREVALENCE
`The reports indicate that smallpoxis more prevalent in the United
`States than it was at this time last year or in 1926. For the week
`ended March 10, 19:38, 42 States reported 1,161 cases of smallpox.
`For the corresponding week of 192? these States reported 876 cases,
`and in 1926 they reported 864 cases for the week. "
`In 1925 the peak of the smallpox curve was reached about the
`middle of January; in 1926 the greatest number of cases was re-
`ported during the second week of February, and in 1927 the peak
`came after the middle of M nreh.
`Some decrease in the incidence of
`smallpox is to be expected, but health officers should be constantly
`on the alert.
`V
`
`A FURTHER STUDY OF EXPERIMENTAL BLACKTONGUE
`WITH SPECIAL REFERENCE TO THE BLACKTONGUE
`PREVENTIVE IN YEAST
`
`By JOSEPH GOLDBERGER and G. A. WHEELER, Surgeons, and R. D. LILLIE and
`L. M. ROGERS, Passed Assistant Surgeons, United States Public Health Service
`
`
`CONTENTS
`
`Introduction. _____________________________ __
`Methods __________________________________ __
`Blacktongue-producing potency of a basic
`experimental diet and of three 01' its modifi-
`cations:
`Basic diet No. 123 _____________________ ._
`Basic diet No. 200. .
`
`Basic diet No. 195. ____
`Basic diet No. 258 _____________________ __
`
`Page
`657
`658
`
`The blacktongue preventive:
`Dried brewers' yeast__________________ ..
`Xeast extract powder__________________ __
`Seidell‘s activated solid________________ __
`Yeast charcoal _________________________ __
`P—P activated fuller’s earth ___________ ._
`Seven and a half hour autoclaved yeast._
`Identity ol the blacktongue preventive. __.__
`Summary and conclusions _________________ _.
`References“.-. . .. ..-_____.._._.___-..--.__.
`Tables .................................... ..
`
`
`659
`661
`663
`664
`
`Page
`
`667
`672
`G74
`677
`681
`683
`684
`688
`680
`600
`
`in a previous emnniunieutien, G-oldherger and Wheeler (1] pre—
`sen ted some. of the results of a study of the prehlem of an experimental
`animal for pelingm. They reported the protlmrtien in the dog,
`
`
`
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`Elysium Health Exhibit 1013
`Page 3 of 22
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`

`March 23,1923
`
`658
`
`of this experimental disease. to human pcllagra was pointed out, and
`was considered so striking as to bc well~nigh conclusive of the identity
`of the two conditions, This view was supported, they behaved, by
`the suggestion of a common etiology indicated by the successful
`production of
`the condition (blacktcnguc) in the dog by feeding
`with pcllagra-producing diets.
`In the prcscnt paper we dcsirc to
`report some further results of the same study.
`METHODS
`
`Our methods of harming, caring for, and feeding ihc experimental
`animals have been described in a preceding communication (1).
`It
`may be noted that tho experimental ration was offered each animal
`once a (lay and, as a rule, freshly prepared. The amount offered
`was, in general, all that the dog; would eat. At first, to insure this,
`the attempt was made to offer an excess; but in order to minimize
`waste this practice was, after a time, modified so that, except for a
`young growing animal, only enough food for the maintenance of
`normal body weight was offered. The animals were weighed once
`a week.
`It may be here recalled, too, that it has been our ' ractice
`to use our dogs repeatedly. Depending on the purpose of tbé experi-
`ment, the animal was or was not reconditioned by a period of stock
`diet feeding between successive experiments, for which purpose our
`stock diet No. 156 (1) was the one most frequently used.
`In what
`follox's, details of this nature will be noted in connection with the
`presentation of the pertinent parts of the history of each experimental
`animal. The diagnostic criteria of experimental blacktongue have
`bccn presented in a preceding article (1). Here we will say only
`that the earliest distinctive buccal lesions are a vivid red injection
`of the floor of the mouth or an erythcma of the mucosa of the upper
`lip in the form of bilaterally symmetrical patches or both.
`
`BLACKTONG-UE-PRODUCING POTENCY OF A BASIC EXPERIMENTAL
`DIET AND OF CERTAIN OF ITS MODIFICATIONS
`
`The composition of the diet with which Goldbcrger and Wheeler
`(1) first successfully induced experimental blacktongue is shown in
`Table 1. This diet was constructed as a somewhat simplified replica
`of the type of diet- found associated with, and, unless suitably sup-
`plcmcnted, believed to be responsible for, the occurrence of pcllagra.
`After a number of modifications of the diet shown in Table 1 had
`been tested, the diet No. 123, shown in Table .2, was constructed and
`subsequently used as the basic blacktongue-producing diet in many
`of our experiments.
`It seems desirable, therefore, to summarize Our
`
`
`
`
`
`
`
`March 23, 1923
`
`
`r609
`
`Basic Diet N0_ 123
`EXPERIMENT 1
`
`
`
`
`
`
`The blacktongue—producing potency of diet No. 123 (Table 2)
`has. been tested in a cons1derable number of animals and under
`J
`7
`various conditions. We shall consider here only those tests however
`that were carried out either in animals after a period of reconditionino'
`following a previous experiment and thus presumably in norlnaol
`condition, or in such as were not previously subjected to experiment
`and were not known to have ever suffered from biacktongue. Such
`tests. were performed in a total of 14 dogs,
`the significant details
`rclatmg to each of which are as follows:
`
`Do(g'713.iMalc. Acquired April 7, 1923, between which date and November
`18, 1.).46, served 111 several experiments and suffered seven attacks of experimental
`blacktongue, the latest of which began Jilly 31, 1925. Was on stock diet N0
`-
`156 for reconditioning from November 18 to December S, 1926
`December 8
`1 26:
`'
`'
`-
`'
`'
`'
`‘
`'
`N0. 123.
`,
`9
`In good condition, weight, 10.1 kilograms; begins test dict
`On January 25, 192:7,lat the end of a period of 48 days, noted the first signs of
`blacktongue,1 an-1niection of the floor of the mouth. Weight 9.4 kilograms
`Confirmatory evrdence of blacktonguc noth on February 10, 1927. Course of
`the attack wasremittent or relapsing in character, accompanied by self-imposed
`partial starvation. Killed with illuminating gas April 28, 1927.
`orDog. 1.1.—Ma-le. Acquired April 7, 1923. Between that date and September
`{0, 192.3, scgg'cd in two experiments and suffered a relapsing attack of blacktono'ue
`)cgmmng
`ay 18, 1923.
`011 stock dict N0. 34C for mo 1'
`'
`'
`'
`'6
`tcmbcr 25 to October 16, 1923.
`J
`L m momng hem EM)-
`Octobcr 16
`1923: I
`-
`'
`'
`'
`'
`'
`'
`‘
`"
`NO‘ 123'
`,
`11 good condition, ue1ght, 9.3 kilograms; begins test diet
`On October 30, 1923, at the end of a period of 14 days noted the first signs of
`blacktonguc, an 1n]cctIon of the floor of the mouth and a. patch of redness on
`thelinucosa of the upper lip opposite the base of the canine tooth on each side
`.
`\Ve1ght, 9.2 kllograins. Further history not relevant.
`0 Dog 15.—l\Iaic. Acquired April 1-1, 1923. Between that date and September
`-5, 192.3, served in two experiments and suffered a relapsing attack of blacktongue
`beginning August 7, 1923.
`011 stock dict No. 34C, for reconditioning from
`September 25 to October 16, 1923.
`'
`October 16
`192°: I
`'
`1'
`'
`v
`'
`I
`"
`'
`NO. 123.,
`o
`11 good condition,
`\\ eight, 9.1 kilograms; begins test diet
`On October 27, 1923, at the end of a period of 11 days, noted the first signs of
`blacktonguc. an injection of the floor of the mouth and slight reddening of the
`
`131100513. of {lilo upperhp.
`\Veight, 9.3 kilograms. Further history not rcluvant
`'
`00 50.
`l‘ ale. Acquired September 25 192”. O "t "-k U *'
`{1“}
`“V
`OCtObcr 16) 1923'
`,
`o
`11 s cc
`dict No. 34C to
`Octobc
`16 D2 :
`'
`"'
`'
`'
`'
`'
`NO. $23.,
`.) 3
`In good condition, “eight, 9 kilograms: begins test dict
`On October 27, 1923, at the end of a pcriod of 11 days, noted the first signs of
`blacktonguc, a definite reddening of
`the floor of the mouth. Wciglii
`3 S
`kilograms. Further history not relevant.
`[A l
`
`’
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`
`Elysium Health Exhibit 1013
`Page 4 of 22
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`

`

`March 23, 1028
`
`March 23, 1928
`
`660
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`661
`
`Dag 51.——Bitch. Acquired September 25, 1923. On stock diet No. 340 to
`October 13 1923.
`h
`l
`V
`g
`i
`p
`Y
`October 18: 1923: In good condition; weight, 7.3 kilograms; begins test diet l\.o.
`123.
`.
`On October 30, 1923, at end of a period of 12 days, noted first Signs of blacktolngue,
`an injection of the floor of the mouth. Weight, 7.8 kilograms. Further history
`not relevant.
`.
`L d. t
`Dog 5.3.—Bitch. Acquired September 25, 1923. On a miscellaneous StOCh
`ie
`to October 22 1923.
`.
`p
`I
`October 22, 1923: In good condition; weight, 8.5 kilograms; begins test diet No.
`123.
`L
`I
`‘
`On November 27, 1923, at the end of a. period of 36 days, noted the first Signs of
`blacktonguc, an injection of the floor of the mouth. Weight, 8.5 kilogiams.
`
`Further history not relevant.
`I
`I
`k 1' t
`Dog 55‘. Bitch. Acquired September 25. 1923. On a miscellaneous stoc.
`( ie
`to October 22 1923.
`I
`.
`October 22, 1923: In good condition; weight, 5.7 kilograms; begins test diet No.
`123.
`'
`
`E
`O
`November 20, 1923: Weight, 5.8 kilograms.
`On November 27, 1923, at the end of a period of 36 days, notco the first Signs 0
`blacktongue, an injection of the floor of the mouth and of the mucosa of the.
`cheeks. Further history not relevant.
`I
`V
`i t
`Dog 5.4.—Bitch. Acquired September 25, 1923. On a miscellaneous stock die
`to October 22, 1923.
`1H1 N
`'
`I
`'
`October 22, 1923: In good condition; weight, 6.2 kilograms; begins test ( ie
`0.
`123.
`f l l
`k
`'
`On November 28, 1923, at end of a period of 37 days, noted the iirst Signs 0
`) ac -
`tongue, an injection of the floor of the mouth. Weight, 6.7 kilograms. Further
`history not relevant.
`I
`_
`r
`n]
`‘
`Dog 57.—Ma1e. Acquired September 25, 1923. On stock dict No. 340 to
`October 16 1923.
`.
`'
`I
`October 16: 1923: In good condition; weight, 7.7 kilograms; begins test diet No.
`123.
`.
`‘
`On November 2, 1923, at the end of a period of 17 days, noted the first Signs of
`blacktongue, an injection of the floor of the mouth.
`\Veight, 8.8 kilograms.
`Further history not relevant.
`L
`.
`V
`a]
`Dog 58.4Bitch. Acquired September 27, 1923. On SLOC'k diet l\o. 040 to
`October 16 1923.
`'
`i
`N
`.
`October 16,, 1923: In good condition; weight, 13. 5 kilograms; begins test diet No.
`12”.
`L
`IL I.
`On November 1, 1923, at the end of a period of 16 days, noted the first signs of
`blacktonguc, an injection of the floor of the mouth. Weight, 14.3 kilograms.
`Further history not relevant.
`.
`‘ A
`v
`M
`Dog 7'9.——Bitcli. Acquired June 9, 1924. On stock (lle No. 100 to December
`i
`-‘
`i
`‘
`16 1994 during which period was bred and whelped a litter 01 five pups which
`were successfully weaned.
`-
`I
`I
`.
`7‘
`.
`L
`I
`December 16, 1924: In good condition; weight, o.3 kilograms; ocgins test dict N0.
`123.
`r 1 ‘1
`'
`h
`‘
`V
`F
`January 27 1925: Weight, 5..)
`(1 ogranis.‘
`On February 7, 1925, at the end of a period of 03 days, noted tne fiist signs of
`
`
`
`
`
`November 21, 1924: I11 good condition; begins test diet N0. 123.
`November 25, 1924: Weight, 9.4 kilograms.
`011 December 27, 1924, at the end of a period of 36 days, noted the first signs of
`blacktongue, a reddening of the floor of the mouth and of the mucosa of the
`checks. Weight, 10.4 kilograms. Further history not relevant.
`Dog 8:3.—Bitch. Acquired October 13, 1924.
`011 stock diet No. 156 to
`November 21, 1924.
`November 21, 1924: In good condition; weight 10.7 kilograms; begins test diet
`No. 123.
`
`On December 23, 1924, at the end of a period of 32 days, holed the first signs of
`blacktongue, reddened bands 011 mucosa of the upper lip oiieueh side; reddening
`of mucosa of the cheeks, reddening of the floor of the mouth. Weight 11.5
`kilograms.
`Treated and used in a study of tomato juice from January 3, 1925, to November
`18, 1926, during which period she had an attack of blacktongue beginning March
`8, 1925.
`
`On stook diet No. 156 for reconditioning from November 18, 1926, to December
`8, 1926. Uri December 7, 1926, weighed 14.8 kilograms.
`December 3, 1926: In good condition; begins a second period of test diet No. 123.
`December 14. 1926: Weight, 13.9 kilograms.
`January 18, 1927: Weight. 13.9 kih‘igrains.
`{In January 22, 192?,
`:lt- the end of a period of 45 days, noted the first signs of
`blackt-onguc, reddened patches; on mucosa of the upper lip in region of canine
`teeth of each side; reddening of mucosa of. the cheek, and reddening of the
`floor of the mouth.
`A rapidly progressive attack, untreated, terminating in
`deal-ii January 29730, 1927.
`Dog 109.——Biteh. Acquired October 29, 1926.
`December 8, 1926.
`Dereliibcr R, 1926: In good condition; weight,
`December 28, 191-36: Weight, 7.9 kiiogrmns.
`On December Jill, 1926, at the end of a period of 22 days,
`riot-ed the first. signs of
`lilzwlitongne, an injection of the ilnnr of the month. The attack took a slmi'ly
`prngrcassive. rnniiilent or relapriing course. acron'ipnnied by self-imposed partial
`starvation; untreated.
`tcrininaioil
`in death April 20, 1927. Weight- on day
`preceding death, 3.3 kilograms.
`Thin.. all 14 of
`the test animals developed blacktongnc, the first
`distinctive signs of which appeal-ml within not to exceed 53 (hin after
`beginning the test diet. Allowed to Lukeitsconrscwithont therapeutic
`in tru'l‘oi‘ehiiwi in two of the dogs, it ended in the death of both iilliii'lals.
`(Inc of the nniiiinls; (dog 32) was subjectcc'l
`to the. test during two
`widely oepariited periods: and responded with liliickti'inguc beginning
`at the end of 3:3 days in the first and at the. and of 45 days in the
`second period.
`
`7.1 kilograms; begins diet N0. 123.
`
`011 stock diet No. 156 to
`
`Basie Diet No. 209
`
`"
`
`In diet No. 123 (Table 2) the principal component, quail[-ital‘ivcly,
`is white corn meal, which forms 66 per cent of the. weight of the dry
`
`Elysium Health Exhibit 1013
`Page 5 of 22
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`

`

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`
`
`March 23, 1923
`
`662
`
`No. 123 of yellow corn meal for the white would have on the black-
`tonguc-producing potency of that diet. Accordingly the following
`test was made:
`
`EXPERIMENT 2
`
`Six dogs, numbered ~14, 52, 60, 63, 64, and 73, were each offered a.
`suit-able daily portion of diet No. 209, the composition of which is
`shown in Table 3. By comparison with Table 2 it may be seen
`that this differs from diet No. 11-33 only in that yellow corn meal is
`usedinstcad of white. The significant facts in relation to each of
`the test animals are briefly as followo:
`Dag é-fi.—Mnle.
`thlpml in the laboratory June 26, 1923. Up to J1113‘ 1?,
`1925. served in a number of feeding experiments and suffered two attacks of
`blacktongue,
`tho bites! of which began December 27, 1924.
`011 stock dict
`No. 156 for reconditioning from July 17', 1925, to August 11, 1925.
`August 11, 1925: In good condition; weight, 12.9 kilograms; begins 1cst dict
`No. 2119.
`
`t signs
`On September 1, 1925, at the end of a period or 21 days, noted the
`of blncklongue, a reddening of the mucosa of the upper lip and afijection
`of the floor of the mouth. Weight, 13.3 kilograms. Further history not
`relevant.
`
`Dag mar—Bitch. Acquired September 25, 1923. Between that date gml
`Jlily 17, 1925, served in several experiments and had four a l' tacks of l‘llfl(rki-{}ngun,
`the. latest of which began March 22-1, 1925.
`(in stock dict No. 159 for recondition-
`ing from July 1?, 1925, to August-11, 1925.
`August 11, 1926: In good condition; weight, 10.8 kilograms; begins test diet
`No.209.
`
`On October 6, 1926, at the end of a period of 56 days, noted the first signs of
`blac-ktomgue, an injection of the floor of the mouth.
`\Veight, 9.8 kilograms:
`Further history nor relevant.
`Dag Gar—Male.
`thlpcd in the laboratory November 4, 1923. Up to July
`17, 11-125, served in a feeding experiment and suffered an attack of blacktmrgue,
`which began July 1-1, 1025.
`(in stock diet No. 156 for reconditioning from
`July 17, 1925, to August 11. 1925.
`August 11, 1925: In good condition; weight, 7.2 kilograms; begins test diet
`N0. 209.
`
`On September 13, 1925, at the end of a. period of 33 days, noted the first signs
`of- blacktonguc, an injection of the floor of the mouth and a flushing of the
`mucosa. of the choc ks. Weight- on September 15, 1925, 7.4 kilograms. Further
`history not relevant.
`Dog (7'3.—«l\'lalc.
`thlped in llic laboratory Noventh 4, 1923. Up to
`July 17, 1935, sci-rm] in two experiments and sufl'ererl two attacks of bhuaktongue,
`the later one of which began March 5, 1925.
`(in stock diet No. 156 for recon-
`ditioning from July 17, 1925, to August 11, 11125.
`August 11, 1925:1[1 good condition; weight, 7.]. kilograms; begins 'l-Pst dirt
`N0. 269.
`
`On August 29, 1925, at the end of a period of 18 days, noted the first signs of
`blacktoogue, a reddening of the floor of the mouth. Weight, 7.5 kilograms
`
`
`
`663
`
`March 23, 1928
`
`which was in evidence June 27, 1925. On stock diet No. 156 for reconditioning
`from July 17 to August 11, 1925.
`August 11, 1925: In good condition; weight, 6.6 kilograms; begins test diet
`No. 209.
`On September 12, 1925, at the end of a. period of 32 days, nol'cd first signs of
`blacktongue, a reddening of the faucial pillars,
`the pharynx, and the floor
`of mouth. Weight, 6.7 kilograms. Further history not relevant.
`Dog 73.-—Bitch. Acquired March 19, 1924. Up to July 17, 1925, suffered
`two uncertain, evanescent attacks of blacktongue, the second of which was in
`evidence July 12, 1925.
`011 stock diet No. 156 for reconditioning from July 17
`to August 11, 1925.
`August 11, 1925: In good condition; weight, 10.7 kilograms; begins test diet
`N0. 209.
`
`On December 17, 1925, at the end of a period of 128 days, noted the first signs
`of blacktongue, a reddening of the mucosa. of the upper lip and of that of the
`cheeks. Weight, 8.3 kilograms. Further history not relevant.
`All six dogs developci'l hlacktongue, the first distinctive signs of
`which appeared within not
`to exceed 56 days after beginning the.
`test diet in five, and at the end of a period of 128 days in one of them.
`It would appear,
`then, that the substitution of yellow for white
`corn meal made no significant difierence in the blacktonguc-produc-
`ing potency of the test diet.
`
`Basic Diet No. 195
`
`In diet No. 123 and its modification, diet No. 209, sodium chloride
`and calcium carbonate are used to supplement the inorganic, “ash,”
`constituents furnished by the other components of
`the ration.
`It
`was thought desirable to determine what effect, if any, on the black-
`tongue-producing potency of diet No. 123 would be produced by the
`substitution of a “complete” salt mixture for the sodium chloride
`and the calcium carbonate. For that purpose experiment 3 was
`carried out.
`
`EXPERIMENT 3
`
`Fire dogs, numbered 9, ‘29, 83, 84, and 85, ware each offered a. suit-
`able daily portion of diet No. 195, the composition of which is shown
`in Table. 4. As may be seen by reference to Table 2, this differs from
`dici- No. 123 only in that the well known “complete” salt mixture
`of Osborne and Mendel (3) replaces the sodium chloride and calcium
`carbonate of the latter diet. The significant facts in relation to each
`of the test animals were, in brief, as follows:
`Dog 9.—l\lalc. Acquired April 1, 1923, between which date and ngruary 28,
`1925, served in a number of experiments and suffered seven attacks of black-
`tongue, the latest of which began February 24, 1935.
`011 stock diet No. 156
`
`
`
`Elysium Health Exhibit 1013
`Page 6 of 22
`
`

`

`
`
`March 23,1928
`
`664
`
`On May 31, 1925, at the end of a period of 60 days, noted the first signs of black-
`tongue, a reddening of the mucosa of the upper lip in the region of the canine
`teeth of each side, and an injection of the floor of the mouth. Weight, 11.4
`kilograms. Further history not relevant.
`Dog 29.—Malc. Acquired May 9, 1923, between which date and February 28,
`1925, served in
`number of experiments and eull'crotl nine attacks of blilckt-migue,
`the latest of which began I"E'lJrIIa-l‘,}' 6, 1925. 0:: stock dict No. 156 for recondition-
`ing from February 28, 1925, to April 1, [925.
`April 1, 1925: In good condition; Weight, 11.7 kilograms; begins diet No. 195.
`On. April 17, 1925, at end of a period of .16 days, not-ed the first signs of. black-
`tongue, a reddening of the mucosa of the upper lip and of the floor of the mouth.
`Weight, 11.1 kilograms. Further history not relevant.
`Dog SJ.~Bitch. Acquired February 20, 1925, from which date to April 1,
`1925, was on stock diet No. 156.
`April 1, 1925: In good condition; weight, 6.8 kilograms; begins test diet No. 195.
`On April 18, 1925, at the end of a period of 17 (lays, noted the first signs of black-
`.
`tongue, a reddening of the floor of the mouth. Weight, 7.3 kilograms. Further
`history nor relevant.
`' Dog 84.—Male. Acquired March 2, 1925, from which date to April 1, 1925,
`was on stock diet No. 156.
`
`April 1, 1925: In good condition; weight, 7.8 kilograms; begins test diet N0. 195.
`On April 23, at the end of a. period of 22 days, noted the first signs of blacktongne,
`rcddened patches on mucosa of the upper lip opposite the canine teeth of
`each side. Weight, 9.1 kilograms. Further history not relevant.
`Dog 85,—Male. Acquired March 2, 1925, from which date to April 1, 1925,
`was on stock diet No. 156.
`
`April 1, 1925: In good condition; weight, 6.9 kilograms; begins test diet N0. 195.
`On April 16, 1925, at the end of a period of 15 days, noted the first signs of black-
`tongue, reddening of mucosa of the upper lip, of that of the cheeks, and of
`the floor of the mouth. Weight, 6.8 kilograms. Further history not relevant.
`I All five test animals developed blacktongue,
`the first distinctive
`signs of which appeared within not to exceed 60 days afterbeginning
`the test diet.
`I
`'
`
`The substitution of the “complete” salt mixture for the sodium
`chloride and calcium carbonate of diet No. 123 made, thus, no sig-
`nificant difference in its blacktongue-producing potency.
`Basic Diet No. 268
`
`Although diet No. 123 (and its modifications, No. 209 and No.
`195) contains a considerable amount of protein, fully one—half of
`which is from casein, it was thought, nevertheless, that the biological
`quality of the protein mixture might, perhaps, be improved by increas-
`ing the casein component, a view that was strengthened by the. results
`of some growth tests in rats (unpublished).
`It seemed desirable,
`therefore, to determine what effect, if any, an increase in the casein
`component of diet No. 123, and thus, presumably, an improvement
`
`
`
`665
`
`EXPERIMENT 4
`
`Mai-ch 23, 1928
`
`Eight dogs, numbered 62, 69, and 101 to 106, inclusive, were each
`offered a suitable daily portion of diet No. 268, the composition of
`which is shown in Table 5. As may be seen, this diet differs from
`No. 123 (Table 2) in containing 50 per cent more casein; it differs
`from No. 123 also in that it contains a “complete” salt mixture in
`the. place of sodium chloride and calcium carbonate. This experiment
`may be considered, therefore, as n. test of the effect not only of a.
`substantial increase in protein, but also of the- use. at the same time,
`of a'complete salt mixture on the hieektonguc-produeing potency of
`that diet.
`
`In this courier-hon, it may he stated that this experiment was
`planned to serve one other purpose—namely, to secure tissues suit-
`able for histopnthological study. Believing that diet No. 268 was
`more nearly complete——at least for maintenance—than N o. 123 or
`its other modifications with respect to all known dietary essentials
`except the specific factor or factors related to blacktongue} it was
`thought that animals fed with it when killed at suitable intervals
`would furnish tissues which might show a sequence of histopatho—
`logical changes which would thus be relatable to the specific dietary
`defect.
`
`The significant facts in relation to each of the test animals, except
`as concerns the histopathology, which will form the subject of a
`separate report by Dr. James Denton, are briefly as follows:
`Dog 6'2.—Male. Whelped in the laboratory November 4, 1923. Up to May 26,
`1926, had one evanescent attack of blacktongue which was in evidence March
`24—27, 1925.
`011 stock dict No. 156 May 26 to June 12, 1926.
`June 12, 1926: In good condition: weight, 8.8 kilograms; begins test diet No. 268.
`On June 26, 1926, at the end of {1. period of 14 days, killed with illuminating gas
`for histepathclogicnl study. He presented no signs of blacktongue. Weight,
`SIS-kilograms.
`Dag 6‘9.——Malc. Whelped in the laboratory November 25, 1923. Up to May
`26, 192i}, served in two experiments. but showed no evidence of blacktongue.
`On stock dict No. 1561mm May 2!} to June 12, 1926.
`June )2, 1936: In good condition; weight, 8.8 kilograms; begins test diet No. 268.
`July 27, 1926: Weight, 8.7 kilograms.
`-
`July 2‘3, 1926. at the end of a period 01316 days, noted first signs of blacktongue,
`a slight injection of the floor of the month which became more definite .‘on
`August 6, and on the following day (August 7) there was noted in addition a
`slight reddening of the mucosa of the cheeks and of the upper lip in the region
`of the canines.
`
`Animal killed with illuminating gas for histopathological study on August 7,
`1926.
`
`Elysium Health Exhibit 1013
`Page 7 of 22
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`March 23, 1928
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`666
`
`June 12, 1926: In good condition; weight, 7.7 kilograms; begins test diet No. 268.
`On July 10, 1926, at the end of a period of 28 days, noted first signs of black-
`tongne, an injection of the floor of the mouth. Weight, 7.9 kilograms. Attack
`slowly progressed. Animal killed with illuminating gas for histopathological
`study on August 7, 1926.
`Dog 102.—Male. Whelped in the laboratory December 9, 1925. Reared on
`stock diet No. 156.
`June 12, 1926: In good condition; begins test diet No. 268.
`June 15: Weight, 6.10 kilograms.
`July 6, 1926: Weight, 6.7 kilograms.
`On July 19, 1926, at the end of a period of 28 days, noted first signs of black-
`tongne, an injection of the floor of the, month. Attack progressed rapidly.
`Animal ki|1ed with illuminating gas for histopathologieal study on July 20,1926.
`Dog IOS.——Male. Whelpod in the laboratory December 9, 1925. Reared on
`stock diet No. 1136.
`Jane 12, 1926: In good condition; begins test diet No. 268,
`June. 15: Weight, 6.7 kilograms.
`July 6, 1926: Weight, 6.4 kilograms.
`On July 10, 1926, at the end of a period of 28 days, noted first signs of black-
`tongne, an inject-ion of the. floor of the mouth. Attack was progressive with
`remissions. Animal killed for histopathologioal study August F, 1926.
`Dog 104.—Bitch. Whelped in the laboratory December (I, 1925. Rented on
`stock dict N0. 156.
`
`June 12, 1926: In good condition: begins test diet No. 268.
`June 15: Weight, 5.7 kilograms.
`On July 6, 1921‘), at the end of a. period of 24 days, noted first signs of black-
`tongue, :1 reddening of the floor of the mouth, of the faucial pillars, and of the
`mucosa of the upper lip. Weight, 5.6 kilograms. Animal killed for histopatho-
`logical study July 10, 1926.
`'
`Dog. 105.—Bitch. Whelped in the laboratory December 9, 1925. Real-ed
`on stock diet N0. 156.
`
`June 12, 1926: In good condition,- begins test diet No. 268.
`J one 15: weight, 4.9 kilograms.
`On July 15, 1926, at the end of a period of 33 days, noted first signs of black-
`tongue, an injection of the floor of the mouth. Weight, 5.2 kilograms. Attack
`mild, relapsing. Animal killed August 7, 1926, for histopathological study.
`Dog Ind—Bitch. Whelped in the laboratory December 9, 1925. Reared'r-on
`stock diet No. 156.
`»
`June 12, 1926: In good condition; begins test diet N0. 268.
`June 15:
`\Veight, 4.3 kilograms.
`On June 26, 1926, at the end of a. period of 14 days, killed for histopathological
`study. Had presented no signs of blacktongue. Weight, 4.5 kilograms.
`Of the eight test animals, two were killed at the end of a test period
`0f 14 days for the purpose of histopathologicsl study. Neither ani—
`mal had shown any evidence of blacktongue. The period of obser-
`vation was so brief that these two animals can not properly be con-
`sidered in appraising the blacktongue-producing potency of the diet.
`The remaining six animals all developed blacktongue, the first (lis-
`
`667
`
`March 23, 1928
`
`use of “complete” salt mixture was without significant effect on tho
`hiacktongne—prod[icing potency of that diet as judged by the attack
`rate and duration of the feeding period before the appearance of the
`first distinctive signs of the disease.
`In the foregoing series of tests of the pellegl‘mpmducing potency
`of diet No. 123 and its modifications, :1. total of 33 dogs were used as
`test- animals, and blacktongno was induced in all but two of them.
`The two that escaped Were the two that Were killed at the end of a
`feeding period of 14 days for the purpose of hisropa'thologioal study.
`Disl'egnl'tiing these as not having had a full opportunity for the derel-
`opment of the disease, blacktongnc was induced in every one of the
`other 31 test annuals.
`In catch of two of these the disease was induced
`on two separate occasions, so that 33 separate. attacks of black tongue
`were induced in the 31 animals under r.:onsideration in this series of
`experhnents. These 33 attacks developed at the end of feeding pe-
`riods that varied between 11 and 128 days in duration, but only one
`oi'tho attacks developed at the end of a period of over 61 days.“
`
`THE BLACKTONGUE PREVENTIVE
`
`Dried Brewers’ Yeast
`
`It may be recalled that Goldberger and Wheeler’s first success in
`experinnmtally inducing blacktongue was with a diet that differed
`frmn that previously tried without sum'ess by Goldberg-er and Lake
`in that it included no milk and was not supplemented with’ yeast (.1).
`The difl’erenee in outeomcled no to suspect that this might bc related
`to the diti'erencc in the diets used. There was already reason to believe
`that milk possessed pellegrn-prereotire properties (4) (5) and thus,
`possibly, also blacktongne-prcventivo properties; but, as the amount
`of milk contained in the diet tested by Goldberger and Lake was very
`sn'iull, it seemed improbable that their failure was attributable, at
`least to an important degree, to the. milk. Up to this time we had had
`no suspicion that yeast might possess pollngra~prcrentive properties;
`but in seeking for the explanation of the difi'orence in results under
`consideration it seemed much more likely that,
`if Goldberger and
`Lake’s failure was due to the preventive act-ion of either food,
`this
`failure was due to the action of the yeast rather than to that of the
`milk. Reasoning thus, it was decided to test this View; and after
`some preliminary trials which it seems superlhlous to detail, the fol-
`lowing experiments n'e'rc poi'i'ori'ncd:
`
`EXPERIMENT 5
`
`
`
`
`
`Elysium Health Exhibit 1013
`Page 8 of 22
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`1111111123, 1928
`668
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`
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`
`
`
` March 23, 1928
`
`
`diet No. 155, the composition of which is shown in Table 6, 01' was fed
`separately in gelatine capsules as a supplement to basic diet N0. 123.
`Comparing diet No. 155 (Table 6) with diet No. 123 (Table 2) it
`will be seen that, so far as it is possible to judge, yeast-containing
`test dict. No. 155 differs from basic lllaektongue—produeing diet No.
`123 significantly only in that 5 per cent of the weight of the solids of
`the former consists of dried yeast.
`Five animals, dogs 15, 30, 35, 46, and 50, were used in this experi-
`ment. The essential facts pertaining to each of the test animals are
`presented in the following:
`
`Dog I5.—Male. Acquired April 14, 1923. By reference to experiment 1 it
`will be seen that this dog developed an attack of blacktongue 011 diet N0. 123,
`the first signs of which were noted on October 27, 1923. On October 30 treat-
`ment was begun by supplementing diet N0. 123 with 5.4 grams (approximately
`1 per cent of the dry ingredients of diet N0. 123) of the dry brewery yeast. Not-
`withstanding this, however, a relapse of
`the signs of blacktongue appeared
`November 24, 1923. Accordingly, 011 November 26 the amount of yeast there-
`tofore included in diet N0. 123 was doubled. On Decclnber 27, 1923, a small
`further increase in the yeast was made, and on March 11, 1924, a still further
`incrcase was made which, after slight compensatory adjustment, made dict
`No. 155.
`
`The evidence of the relapse noted 011 November 24, 1923, faded in a few days
`and thereafter to November 21, 1924, the dog continued in good condition without
`presenting recognizable evidence of blacktonguc.
`.
`On November 21, 1924, yeast-containing test diet No. 155