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A STUDY OF THE EFFECY OF
FEEDING T‘HYROPROTEIN TO
LAYING WIRES OF ONE STRAIN
OF RHCDDE ESLAND REDS

Thais for The Degree cf M, S.
MIG-{36AM STATE CGLLEGE

Maurice 5. Armsi'rang
1947

THESIS

This is to certifg that the {

thesis entitled

A Study of the Effect of Feeding Thyroprotein
To Laying Birds of One Strain of _
Rhode Island Reds y

presented by

Eeurice S. Armstrong

has been accepted towards fulfillment

of the requirements for

Master Omngegree in_P_°_Lll-ULV- .HUSbandrY

 

Major professor

Date Decembe r'_1_§1_;947

 

M-795

 

A S'NDY OF Tl! MOT OF FEEDING THYROROTIIN 1'0 LAYING BIRDS
OF M SMIN CE RHODB ISLAND REDS

B!
maria. S . Armstrong
1’

A THISIS
Submittod to the School of Groduoto Studio: of mohim
suto College of Agriculture and Applied Scionoo
in partial £111!th of tho roquirononts
for tho than of
(mm tr some)
Doparhnont of Poultry mm

1947

THESIS

ACKNOWLEDGE”?

Tho author mm to apron his opprooiotion to luocioto
Protoooor J’. l. Boudoon Ind Professor c. 6. card of tho Poultry
Donortnont, out Doctor n. P. 331mb of tho Physiology nopmnt of
[10111911 Stoto College, for thoir odvioo, miotmoo, and cooperation

1‘ m ”rko

194.381

Table of Contents

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Liuratm Rm.‘ 0 O O O O O O O O O O O O O O O O O O O O O O

A.

B.

P.

G.

Effect of Feeding Rev or Deoicceted Thyroid on the Egg
Mduction Of rule 0 O O O O O O O O O O I O O O O O O 0

Influence of Thyroidectony on the Egg Production of Poole.
Relation of Thyroid Function to Molt end Feather Growth. .

leeccietion of Egg Production, Seneecenoe, and Seasonal
CycleenithThyroidFunction. . . . . . . . . . . . . . .

DeveIOpment end Syntheeie of Thyroprotein . . . . . . . .

Effect of Thyroprotein on the Rate of Growth and Feather-
lngofBabyChlcks....................

Effect of Feeding Thyroprotein on the Egg Production of
“Ying ’0'18 0 O O O O 0 O O O O 0 O O 0 O O O 0 O O O O O

Reeeone for Undertaking Experiment . . . . . . . . . . . . . . .

Materials and Experimental Procedure . . . . . . . . . . . . . .

A.

Basic for Selection of Pullete . . . . . . . . . . . . . .
Experimental Procedure for Pullete . . . . . . . . . . . .
Experimental Procedure for Too and Three Year Old Birds. .

Experimental Procedure for Testing for the Effect of
Thyroprotein on Egg Shell Quelity . . . . . . . . . . . .

1. Experiments Relating to the Effect of Thyroid Materiel
OnEgsShcllQutllty...oo............

2. Experiments Involving the Verioue Teete for Egg Shell
QUfility o o o o e e e e e e e o e o e e e e o e e e e

3. Reasone for Employing Moisture Lose Teet . . . . . . .
4. Egg Shell Quality Tents and Recorde . . . . . . . . .

Experimental Procedure to 'Deet for the Effect of Iodineted
lenonlggWolghts.o.........o.....o

Page

10
10

13

13

14
14

15

15

hwimnmRMIuoeoeeeeeeeeeee

‘o P1111.“ " Egg ROdUCtion e e e e e e e e e

B. Pullete -

1c. Pullets-Molt..............
D. Pulleto-Egg Shell Quality . . . . . . .

E. Pullete-EggWeighta............

F. The and Three Year Old Birds
0. Two and

Discussion.....oo..............

‘e ESSPPOdUCtioneeeeeeeeee

B. Body Weight and Feed Consumption .

0. volt.........’......

D. Egg Shell Quality and Egg Weights.

Sunry

Oooooooooooooooo

Bibliography..............

Body Weight and Feed Consumption

Egg Production

Three Year Old Birds - Peed Consumption

Page
17

17

23
23
27

29

36
56
38
39
39
41

43

A STUDY OF '11-: NOT OI" FEEDING THYROPRO‘I‘EIN ‘10 LAYING BIRDS
OF ONE 8mm OF BHODB ISL-AID REDS

Introduction

The process of reproduction of the fool is highly complicated
and involves considerable glandular activity in the body. The complete
physiological processes have not as yet been vorked out, hovever a
great deal of infomtion pertaining to then he been obtained. mug
the more inpertant of the glands associatedsith egg production is the
thyroid. In the chicken the thyroid is a small, oval red body located
on the ventral side of the comet: carotid artery at the point where it
touches the jugular vein. The thyroid gland, Ihich is protein in
nature, secretes the hormone, thyroxine, shich is an iodinecontaining
derivative of the anino acid, tyrosine. Thyroxine is thought to be
catalytic in tint it raises the excitative capacity of cells, yet
vithstands digestion, Kendall (1929) . The more general influences of
thyroxine are that it is responsible for the aaintenance of nornl
netabolism and alterations in the metabolic rate to counteract changes
in the environment. In addition thyrexine plays an important role in
the promotion of growth, this affect apparently reuniting from its
actiu on metabolism in general, Lorna-n (1942). These general effects
are brought about by the following more specific influences: oxidation
of carbohydrates, and more primarily oxidation of proteins and fats;
heat production; and tissue consumption of oxygen.

Since environmental conditions affect body netsbolisn, it follova
that they similarly influence the thyroid. Thyroid gland activity and

body aetabolisa are decreased by high temperatures and increased by low
temperatures, Kendall (1929). Furthermore it is knovn that thyroid
activity tends to decrease vith advancing age.

Thyroid secretion has a too-fold influence on egg production.
Since egg production is to some extent associated with body metabolism,
it cones under the influence of thyromine in this respect. In addition
to the above effect, an excess of thyroxine in the tissues results in
a depression of the thyrotropio activity and an increase of the gonado-
tropic activity of the anterior pituitary; the gonadotropic hormone in
turn influences ovarian activity, Marine (1940). a deficiency of
thyroxina in the tissues results in the opposite of this, Cameron
(1940). In vies of these facts it vould appear that an optimum level
of thyroxine in the body would be the most conducive to high egg

production.

Literature Review

Effect of Peeding Raw or Deoiccated Thyroid on the Egg
Production of Powls

Various experiments have been performed with the feeding of ran
or desiccated thyroid to fowls and their influence on egg production.
Crew and Huxley (1923) fed raw thyroid to six birds, using six other
birds as controls, but were unable to obtain any significant results
pertaining to egg production. Following this Crew (1925) administered
desiccated thyroid to seven senile hens. All birds promptly melted,
the new plumage being characteristic of younger fowls. In addition an
enlargement and reddening of head furnishings took place. The low egg
production of d. 67 eggs per hen for six months preceding the experiment
increased to thirty-four eggs during the treatment, and twenty-four
during the six months following the treatment. Desiccated thyroid
(one mg. of thyroid iodine per 1750 gm. of body weight) was administered
to laying hens by lsmundson and Pinsky (1935) with the following results:
slight increase in the amount of shell, a reduction of yolk weight and

rate of ovum growth, and a decrease in body weight and egg production.

Influence of Thyroidectony on the Egg Production of Powls

Winchester (1939) decreased the egg production of White Leghorn
hens eighty-five percent by thyroidectomy. leekly injections of thyro-
xine resulted in an increase of the egg production of these same fowls
to forty and fifty to sixty percent of that of nornl fowls. Complete
thyroidectony by laylor and Burneister (1940) brought about a decrease

in egg production of one-third to one-fourth that of normal fowls.
while incompletely thyroidectomiaed birds showed a decline in egg pro-

duction of about two-thirds that of normal.

Relation of Thyroid Function to Holt and Feather Growth

Cele and Reid (192A) and Mercy (1925) found evidence to the
affect that thyroid-fed birds showed more rapid and uniform feather
growth, and that the thyroid plays a role in the regulation of the
growth and melting of feathers. In an experiment conducted by Cole and
Flatt (1928) thyroid-fed birds exhibited a rapid molt, dropping all fea-

thers simultaneously and growing new ones uniformly.

Association of Egg Production, Seascapes, and Seasonal
Cycles with Thyroid Function

Various investigations involving the association of environment,
egg production, and thyroid function have been undertaken. Brody,
tbndmon, and Kempster (1923) and Clark (1940) obtained evidence that
the average annual decline in egg production is twelve to twenty percent
of tint of the preceding year. Indications that egg production is
definitely influenced by light and length of day were obtained by
Bissonnotte ( 1933) and Winchester (1940). Yearling Rhode Island Red
hens were subjected to a fourteen hour day-light period by Byorly and
Gardner (1943). Those birds laid at a rate of fifty-eight percent from
December through larch, while the controls laid at the rate of twenty-
four percent for the some period, being subjected to the normal daylight

periods. The molt was not affected by the extended light period.

Dempsey and Atwood (1943) found evidence which indicated that the
thyroid secretes its active principle more rapidly in cold than in warm
environments, and that the basal metabolic rate is elevated by exposure

to OOMo

Development and Synthesis of Thyroprotein

Considerable work has been done relating to the development and
synthesis of thyroprotein. Ludwig and lutzenbechor (1936 and 1939),
‘ and Reinoke and lurner (1943) were able to isolate thyroxine from
artificially produced iodinated proteins including iodinated casein
(thyroprotein). Reineke and Turner (1942) developed the currently
used method of producing iodinatod casein by treating casein, in the
presence of manganese as acatalyst, with iodine and sodium bicarbonate
in a warm water bath followed by on eighteen to twenty hour incubation
period, and the iodinated casein was precipitated out by the isoelectric
method, following dialysis. The thyroidal activity of an iodinatod
potency attains a maximum when two atom of iodine have been added for
each sole of tyrosine, and that additional iodination results in a
definite decline in thyroidal activity. Thyroactivo proteins have also
been obtained with egg albumin, soybean proteins, silk fibroin, and
serum globulin, but casein has been most widely used due to availability
and ease of mipulation.

The iodinated casein (protamone--supplied by Cerophyl Laboratories,
Inc., 2438 Broadway, Kansas City, llissouri) prepared above tested 2.13
thyroxino by biological essay and 3.03 percent by chemical assay. Bio-

logical assayo have been accomplished in various ways. Reineke and

hoe

 

m: (1942) employed the metamorphosis of tadpoles, and the oxygen
consumption of guinea pigs due to thyroid stimlation as a measure of
the thyroxine level of iodinated proteins. Another sethod of biologi-
cal assay, developed by Miner, Reineke, and Turner (1944), involved
treating day old chicks with thiouracil (anti-thyroid hormone), and
observing the rate at which thyroactive proteins caused a decrease in
the sine of the enlarad thyroids. Reineke, et. a1. (1945) worked out

a chemical assay of iodinated proteins, which consisted of the hydrolysis
of iedocasein with forty percent barium hydroxide solution, the extrac-
tion of the thyroxine with n-butanol, and determination of the iodine

content of the purified extract.

“feet of Thyroprotein on the Rate of Growth and
Feathering of Baby Chicks

although this work is concerned primarily with the egg production
of laying birds, a few experiments are herein reviewed relating to the
rate of growth and feathering of baby chicks, since molt and subsequent
growth of feathers is a normal occurenoe during the sumsr and fall
months of the laying year. Irwin, Reineke, and Turner (1943) fed thyro-
protein at the rate of thirty-six grams per one hundred pounds of feed
to lhite Plymouth Rock chicks, and at the end of twelve weeks the exper-
imental chicks were slightly heavier and showed improved feather growth
as compared with the controls. Rhode Island Red chicks raised to twelve
weeks on rations containing .025 to .08 percent iodinated proteins by
Parker (1943) showed slightly greater gains in body weight than did the
controls. lbrner, Irwin, and Reineke (1944) fed thyrcpretein at the '

rate of forty-five gram per one hundred pounds of feed to Barred

Plymouth Rock «chrels with the result tint the treated birds cow
slightly slower than the controls, indicating tint this amount was the

uppper limit of tolerance without a marked depression of the growth rate.

Effect of Feeding Thyroprotein on the Egg Production
of Laying Powls

The first experiment involving the feeding of thyroprotein to lay-
ing fowls was conducted by Mner, Irwin, and Reineke (1945). Party-
eight two year old lhite Leghorn hens, selected on the basis of physi-
cal characteristics, were divided into four coups with one coup as
controls; the other coups receiving the following levels of thyropro-
tein (2.73 percent thyroxine by biological assay); five, ten, and
twenty cams per one hundred pounds of feed. The average percentac
egg production records obtained for one year were: controls, 22.6;
five can level, 38.1; ten gram level, 40.6; and twenty gram level,
30.7. The five and ten gram level coups attained a higher level of
egg production and maintained the level longer firing the summer,
however the thyroprotein—fed birds dropped precipitously in egg produc-
tion in late August and went into a salt. lo marked differences were
obtained in regard to egg weights, body weight, or mortality.

In a nu:- experiment ms, Kempster, m1, and Reineke (1945)
used the some white Leghorns with some addition. A ten cam levwl of
thyroprotein (2.73 percent thyroxine by biological assay) was fed to
twenty-four of the birds, and another coup of twenty-four birds was '
used as the control coup. In addition ten and twenty can levels of
thyroprotein were fed in the ration to two coups of twelve Rhode Island
Bed pallets each, and a third group received the basal ration with

physical characteristics serving as the basis of selection. ill the
birds were subdected to a.ten and one-half hour daylight period. The
three year old thyroprotein group had an average egg production record
of 38.8 percent during October to lay, 36.3 percent during Hay to
October, with.an overall average of 37.7 percent; while the controls
had an average of 42.0 percent during the first six months, 23.4 per-
cent during the last half of the year, and 33.9 percent for the whole
year. The egg production of the controls decreased 43.8 percent in.the
second half of the year, while that of the experimental group decreased
only 6.4 percent. The three year egg production of those three year
old birds surviving this and the previous experiment.was as follows:
controls, 81.3 percent and 89.5 percent of their previous years' produc-
tion; thyroprotein-fed group, 72.5 percent and 102.3 percent of the
previous years' production. Similar results were obtained with the
pullets. During the first half of the experiment the controls laid.at
the rate of 70.8 percent, the ten gran level at 71.6 percent, and the
twenty gram level at 61.7 percent. In the second half the average egg
production.was as folloss: controls, 52.7 percent, ten gram level,

69.7 percent; and twenty gram level, 51.7 percent. the overall percent-
ages were: for the controls, 62.8 percent; for the ten gram level, 67.2
percent; and for the twenty gram level, 52.0 percent. Decreases of 25.6
percent, 6.1 percent, and 14.8 percent during the final half of the
experiment were recorded by the controls, ten gram level, and the twenty

gram level, respectively.

Reasons fer-Undertaking Experiment

In view of the foregoing information further work involving the
feeding of thyroprotein to laying birds to determine its effect on
egg production is necessary, before its value in this respect can be
definitely established. The significant results obtained by Turner,
Irwin, and Reineke (1945), and mrner, et. a1. (1945) indicate that
thyroprotein has possibilities as a method of increasing egg produc-
tion. Furthermore, there is an ever continuing need to increase egg
production by delaying the seasonal decline, either by delaying molt,
or speeding it up; this applies both to pullets and old hens. There-
fore, it seemed that an experiment, involving the feeding of thyropro-
tein to laying birds, to determine its effect on egg production, would
be of considerable interest. It would appear that this procedure would
provide the birds with a uniform level of thyroxine in the body, parti-
cularly during the time, when the hot weather of simmer results in a
decrease of the body thyroidal activity. As a result it was believed
that a test of this nature would be more apt to be significant if

conducted during the spring, summer, and early fall.

10

laterials and prerinntal Procedure

A. Basic for Selection of Pullets

In order to have the birds employed in this experiment as closely
matched in relation to egg production ability as possible, they were
paired up on the following basis: age at first egg, rate (intensity),
total lay for the first four months of production, and where possible,
the same dam. Out of ninety-six Rhode Island Red pullets, which were
pedigreed from the Michigan State College flock, twenty-seven pairs
were selected with ten days being the greatest variation in age at
first egg, and eight eggs the extreme as to comparison in reference to
total lay, while three of the pairs had the same dam. Following exams
ination of the birds, three pairs of the twenty-seven were discarded,
as one of the birds in each instance was unusually light, and.notice-
ahly dull and inactive. These birds had been reared on range and

housed in laying cages during September and October, 1946.

B. lxperimental Procedure for Pullets

The experiment was started on larch 3, 1947, although the experi-
mental birds received ten grams of thyroprotein per one hundred pounds
of feed in their diet for a period of two weeks prior to the official
starting date. Forty-eight pullets were divided into two coups of
twenty-four birds each, and housed in laying cages. lost of the birds
were in production at the start of the experiment, and none were melt-
ing. The birds were divided into groups I and II with the former behng
the control. These birds had been subjected to a thirteen and one-half

hour daylight period prior to the beginning of the experiment, and this

light period was extended to fourteen hours for the duration of the

experiment. The controls were fed the regular college battery laying

nah (15.0 percent crude protein), which was made up, as follows:
foodstuff Amount (pounds)
Corn meal ............................. 690
Round oats . 4106
Wheat bran ............................ 300
Wheat middlings ....................... 200
Alfalfa meal (17% dehydrated) ......... 60
Heat scraps ........................... 60
Drymilk ..... 40
fish meal ............................. 50
Soybean meal .......................... 50
Oyster shell flour .................... 100
Steamed bone meal .... 30
Salt ..... 12
rich oil (400 A, 2000 D) .............. __8
Total 2000
woup II received the me ration plus ten grams of iodinated casein
(3.04 percent thyroxine by chemical assay) per one hundred pounds of
feed. The ten gram level of thyroxine was selected, because it was
found to be the most optimm in the experiments performed by Turner,
et. a1. (1945).
Inasmuch as the amount of thyroprotein added to the ration was

very snll, it had to be meticulously mixed to insure even distribut

ion.

The ten grams was first thoroughly mixed with one-half pound of feed.

This one-half pound of feed was then mixed with one pound of feed.

This

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process he progressively continued, until ten pounds had been mixed.
The ten pounds mind in the above manner were then added to the remain-
ing ninety pounds, and the final mixing was done with a McClellan
Etch Illnr. ‘

The experiment was divided into four week intervals to facilitate
record keeping. The egg production, and feed consumption records were
tabulated at the end of each four week period. it this same time the
birds were. individually weighed and their respective weights recorded.
These birds were checked for molt on October 5 and 26, and November 9,
1947, on the basis of the number of new primaries present. The work

herein reported was concluded on November 9, 1947.

C. prerimental Procedure for he and Thru Year Old Birds

Since thyroid activity as well as egg production decreases with
age, a study of the influence of feeding thyroprotein on the egg pro-
duction of two and three year old hens was also included in this
experiment. This portion of the experiment was started on Hay 15, 1947.
These birds were pedigreed Rhode Island Reds from the Ilichigan State
College flock and were paired up on the basis of egg production for
the first two and three years, and as near as possible, age at first
egg. The greatest variation in two year old birds for first year pro-
duction was three, for second year production, fifteen, mile the
differences in age at first egg varied from five to seventy-five days.
for the three year old birds the largest difference for the first year
egg production was sixteen, for the second year, twenty-seven, and
for the third year, thirteen. Differences in age at first egg for these

birds ranged from two to forty-nine days.

13

Ihese birds were divided into groups of eight as follows:

group III, two year old controls; group IV, two year old thyroprotein-
fed birds; group V, three year old controls; group VI, three year old
thyroprotein-fed birds. As with the pullets the two and three year old
birds were housed in laying cages and exposed to a fourteen hour day-
light period. Egg production and feed consumption records were tam-
lated at the end of four week intervals. The controls were placed on
the regular college battery laying mash, while the experimental birds
were given the some ration as the experimntal pullets.

D. aperimental Procedure for ibsting for the lffect of Thyropretein

on Egg Shell Quality.

1. Experiments Relating to the Effect of Thyroid Material on
ng Shall Quality.

Involved in the overall effect of thyroxine on body metabolism is
a further influence on calcium metabolism, and consequently a possible
effect on egg shell quality. lsmundson and Pinsky (1935) were able to
bring about a slight increase in the amount of shell by feeding
desiccated thyroid to laying hens. Gutteridge and Pratt (1946) con-
ducted an experiment involving the feeding of vitamins ”2 and D3, mid
iodinated casein at the rate of fifteen grams per one hundred pounds of
feed, to white Leghorn pullets. 0n the basis of the specific gravity
test, the coup receiving the iodocasein showed a significant increase

in egg shell quality compared with those receiving vitamins : D and 03,

2
and the controls. No significant differences in egg production were
recorded for the duration of the experiment, which extended from

January through July. In a later experiment of similar nature Gutteridge

and flovikoff (1947) obtained results which verified the previous work.

13

mass birds were divided into groups of eight as follows:

group III, two year old controls; group IV, two year old thyroprotein-
fed birds; group V, three year old controls; group VI, three year old
thyroprotein-fed birds. As with the pullets the two and three year old
birds were housed in laying cages and exposed to a fourteen hour day-
light period. ngg production and feed consumption records were tabu-
lated at the end of four week intervals. The controls were placed on
the regular college battery laying mash, while the experimental birds
were given the same ration as the experimental pullets.

D. Experimental Procedure for Rating for the lffect of Thyropretein

on lgg Shell Quality.

1. Experiments Relating to the Effect of Thyroid Itaterial on
ng Shell Quality.

Involved in the overall effect of thyroxine on body metabolism is
a further influence on calcium metabolism, and consequently a possible
effect on egg shell quality. Asmundson and Pinsky (1935) were able to
bring about a slight increase in the amount of shell by feeding
desiccated thyroid to laying hens. Gutteridge and Pratt (1946) con-
ducted an experiment involving the feeding of vitamins Ba and 33, and
iodinated casein at the rate of fifteen grams per one hundred pounds of
feed, to White Leghorn pullets. 0n the basis of the specific gravity
test, the group receiving the iodocasein showed a significant increase

in egg shell quality compared with those receiving vitamins - D and D

2 3’

and the controls. No significant differences in egg production were
recorded for the duration of the experiment, which extended from
January through July. In a later experiment of similar nature Gutteridge

and Rovikoff (1947) obtained results which verified the previous work.

.2

14

In this case the thyroprotein was fed at the rate of ten gram per one
hundred pounds of feed, and as before egg production for the- various

groups was about the same.

2. Experiments Involving the Various Tests for Egg Shell Quality

Various nthods have been devised for testing for egg shell
quality. “organ (1932) tested for percent of shell, breaking strength,
and thickness of shell by breaking the shell by applying increased
weight to the egg at right angles to the long axis. A test for breaking
strength of the shell by measuring the striking force of a mass of
known weight falling as a free body through a given distance onto the
egg we devised by Swanson and James- (1932). Lund, Heiman, and wilhelm
(1938) employed a complicated device involving a pulley, fulcrum, car-
riage, weight, and graduated scale, as a means of testing the egg shell
for breaking strength and shell thickness. in experiment using the
moisture loss as a criterion of egg shell quality was conducted by
Quinn, etc. a1. (1945). The egg weight in grams (nearest tenth) was
recorded the day after the eggs were laid, and the eggs were then sub-
Jected to a fourteen day incubation period in a forced-draft incubator
(99.50 r. and sixty percent relative humidity). Following the incuba-

tion period the eggs were re-weighed to ascertain the moisture loss.

3. Reasons for Employing loisture Loss Test

The moisture loss test has the advantage of being a simple total
score test for egg shell quality. It does not involve testing specifi-
cally for such factors as, shell strength, porosity, thickness, percent
of shell, and the shell membrane characteristics. Individual tests for

these. numerous factors tend to bring in such variables, as the manner

15

of breaking or crushing the shell and the fact that different parts

of the egg shell respond differently. Further the size of the air cell
has long been used as one of the standards of quality in eggs, and the
less of moisture, which is accompanied by the less of weight, results
in increased size of the air cell. Though this type of test does

not test individually for the various shell quality factors, it does

serve as an overall measure of these factors.

4. Egg Shell Quality Tests and Records

A record including the results of the incubation of the eggs of
each group for the week of Kay 19 to 23, 1947, was made. The average
moisture loss rates of the two groups of pullets were compared to
determine the effect, if any, of thyroprotein on egg shell quality. In
order to further investigate the possible influence of iodocasein on
egg shell quality, a second test was conducted, which included the
incubation of those eggs laid from October 13 to 20, 1947. The latter
records were summarized individually first, then compared with those~of
the first test to determine whether or not, the feeding of thyroprotein
might cause a seasonal'variation in egg shell quality.
It. hperimontal hocedure' to Test for the Effectlof Iodinated casein

on Egg Heights

In order to obtain an idea of the possible effect of iodocasein on
egg weights, the eggs of the two groups of pullets were weighed once each
week for the first two and one-half months of the experiment, than twice
in June and once in July, and the weighings of the eggs of October I:
to 20 were also included. These weighings were then divided into five

different intervals. The egg weight which was nearest the mean for that

16

particular period for each.bird that laid during that period was recorded.
The average egg weight of each period was then computed. Finally an
overall.amerage was computed. The average egg weights of both groups of

the pullets for the five periods were then compared and summarized.

17

Experimental Results
Pullets-Egg Production

The Rhode Island Red pullets employed in this experiment laid at
their contest rate during the first four week interval, which began
larch 3, 194'! (Tables 1 and 2, and Figure no. 1). This peak performance
by both coups was followed in the second four week period by a decrease
of 0.7 percent in production by the controls, and a decrease of 15.2
percent by the experimental birds. The two coups continued to decrease
throughout the third interval of April 28 to may 26. The controls
declined more rapidly through this third period, but their production
remained above that of the thyroprotein-fed birds. The period from
lay 26 to June 23 found both coups showing an increase in production
at about the same rate.

The experimental birds apparently began their seasonal decline in
late June, while the control birds registered a slight increase in their
production during the period of June 22 to July 21, at which time they
began their seasonal decline. A rapid reduction in production was
recorded by the controls from July 21 to September 15. The thyropro-
tein-fed coup also exhibited a similar decrease during this period,
but at a slightly more retarded rate. Both groups fell off precipitously
in production fnm September 15 to October 13, as a result of their going
into a melt. hiring the last four week interval the experimental birds
increased their production 8.? percent, while the controls increased
only 0.5 percent.

The two coups were nearly equal in egg production on the basis of

their laying performs” during the fir“ five months of the laying

18

year, the controls laying at the rate of 38.3 percent during this
period, with the experimental birds laying at the rate of 37.9 percent.
Except for the first and last intervals the control birds consistently
laid at a greater rate, this rate varying from 4.6 to 20.8 percent, or
at an average of 11.7 percent. The controls had an average egg pro-
duction of 47.8 percent for the first four intervals, while the thyro-
protein-fed birds averaged 41.9 percent for the some period.

The seasonal decline of the controls extended over three four-week
intervals, and their production average for this period was 28.3 percent
with the average decrease being 12.8 percent. In contrast to this the
seasonal decline of the experimental coup covered a duration of four
four-seek intervals, while the average production for this period as
19.8 percent and the average decrease for this period was 8.5 percent.
During the second interval of their seasonal decline, that is, from
July 21 to iugust 17, the emerimental birds showed a tendency to resist
tb decline, by dropping only 2.0 percent in their average woduction,
while ttm controls decreased 7.4 percent during this time. As indicated
by hble 9, this is the period of the highest nan temperature. In
referring to this table it will be noted that these were outside atmos-
pheric temperatures. During March, April, and May, artificial heat was
supplied to maintain a constant temperature of sixty decees rarenheit
in the battery room. Therefore these temperamres listed in the table
after hey are the only ones that apply for this experiment.

Statistical analysis, employing standard deviations and standard
error, showed that the difference in the overall average egg production
0f the two coups of pullets was not large enough to be significant.

The standard deviation of the overall average egg production of the

19

 

 

 

 

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22

control birds was 31.9, while for the thyroprotein-fed birds it was 24.0.
The standard error, using these standard deviations, for the two groups
was 9.16. The difference between the means of the two groups was 21.6,
which was considerably below three times the standard error, or 27.5.

The fact that this difference of 21.6 was more than twice the standard
error, or 18.3, would indicate that it approached significance. However,
it would appear that other factors, such as, variation in the amount of
thyruine available, or environmental conditions, are involved here,

and further work would be necessary to determine whether or not this

difference was definitely significant.

Pallets-Body Height and Feed Consumption

The body seights of the two groups of pullets followed a rather
similar pattern for the duration of the experiment (Tables 3 and 4).
The controls outweighed the experimental group by about 0.45 of a pound
on larch 31. In general, this difference continued throughout the
experiment, except for a difference of 0.3 of a pound in favor of the
controls on dugust 18, and 0.6 of a pound, again in favor of the con-
trols, on October 13. Both youps showed a slight decline for the next
two intervals following the March 31 weighing. A slight increase
occurred from May 26 to June 23, and this was followed by a negligible
drop during the next interval. Pros July 21 to September 15 the two
groups both increased about 0.25 of a pound. Although the therprotein-
fed birds showed a small decrease in bodyrweight from September 15 to
October 13, while the controls remained about the some for this period,
both groups increased a little during the last interval. 0n the basis
of the overall average, the controls weighed 5.38 pounds, and the experi-

mental birds weighed 4.93 pounds.

23

The feed consumption of the two lots of pullets included those
birds that died and their partners. Because of this the number of birds
in the two groups differed beginning with the‘ interval of July 21 to
lugust 18. The fact, that four of the controls died, while only one of
the experimental birds died, would infer that the latter group would
have the greater feed consumption. Acmally the controls consumed the
most feed, although their consumption was not noticeably larger, the

difference ranging from 0.2 to 0.7 of a pound per bird (Tables 3 and 4).

Pullets-llolt

As indicated by Table 5, the ...1. of both groups of the pullets
was well underway when the first check was made. it the tin of the
first observation at October 5, 1947, the thyroprotein-fed birds had
melted 32.2 percent, while the controls had molted 19.7 percent. Buring
the five weeks following October 5, the birds were checked twice, first
on October 26, and again on November 9. Both of these observations
showed the two groups to be melting at about the same rate. he controls
increased 13.7 percent in the progress of their molt during the five

weeks, while the experimental birds showed an increase of 14.5 percent.

Pallets—Egg Shell Quality
The first test for differences in egg shell quality was rude with
the eggs of lay l9 to 23, 1947 (Table 6). The average moisture loss in
grams for the two groups of pullets varied daily from 0.2 to 0.6 of a
gram. The overall average for the controls was 2.85 grams, or 4.8 per-
cent; for the experimental birds it was 3.14 grams, or 5.5 percent.
The next test involving egg shell quality included those eggs of

October 13 to 20, 1947 (Table 7). The daily variations in moisture loss

24

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

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27

in pens for the two groups for these eggs ranged from 0’}2 to 1.2 grams.
The controls had on overall enrege moisture loss of 3.42 grams or 6.1

percent, and the experimental birds had an overall average of 3.43

grams or 6.2 percent.

Pullets—Rgg Weights

On the besis of two weighings from March 11 to March 18, the con-
_ trols had on average egg weight of 58.0 grams, while the experimental
' birds had on everege egg weight of 56.9 grams (Table 8). The average
egg ueight of the controls increased to 59.1 grams for the seighinge of
by 7 to Kay 23, end the final ulghlngs of October 13 to 20 abound I.
decrease to 55.8 grams. The storage egg weight of 58.6 grams recorded
during the weighings of Key 28 to July 21 m the peak for the experi-
ments]. birds, and their lmst everoae, 55.8 grams occurred during the
October 13 to 20 veighings. The controls had an overall average of
57.9 grams, and the experimentel birds had on enroll average of 57.7

Fm,

I .|.Iy

27

in.grams for the two groups for these eggs ranged from d§2 to 1.2 grams.
The controls had an overall average moisture loss of 3.42 grams or 6.1
percent, and the experimental birds had an overall average of 3.43

grams or 6.2 percent.

Pulletsémgg‘Weights
On the basis of two weighings from March 11 to March 18, the con-
_ trols had on average egg weight of 58.0 grams, while the experimental
' birds had an average egg weight of 56.9 grams (Table 8). The average
egg weight of the controls increased to 59.1 grams for the weighings of
In: 7 to [lay 23, end the final weighings of October 13 to 20 shoved e.
decrease to 55.8 grams. The average egg weight of 58.6 grams recorded
during the weighings of May 28 to July 21 was the peak for the experi-
mental birds, end their lowest averaa, 55.8 grams occurred during the
October 13 to 20 weighings. The controls had an overall average of
57.9 grams, and the experimental birds had an overall average of 57.7

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Ivnuaennuun II 0 Hanna

29

The and Three‘Year Old Birds - Egg Production

Both groups of the two year old birds showed a marked drop in egg
production during the second interval which extended from Ilay to June 22
(Tables 10 and u, and Fibure no. 2). 1111s was followed by an increase
of 15.4 percent by the experimental birds, and 10.2 percent for the
controls during the next period. me two groups both started their
seasonal decline at the beginning of the fourth interval on July 21.
Iro- July 21 to lugust 17 the controls dropped 11.8 percent, while the
thyroprotein-fed birds dropped only 0.5 percent for the same period.
The controls decreased 20.9 percent to an average egg production of 5.1
percent from August 18 to October 13. For the same period the experi-
mental birds declined 10.2 percent to an average egg production of 4.6
percent. mring the last interval the controls increased their produc-
tion to 9.2 percent, while the thyroprotein-fed group remained the same.
Except for the interval of September 15 to October 13, the controls
maintained a markedly higher average egg production than did the experi-
mental birds. This superiority ranged from 4.6 to 22.5 percent. In
addition three of the controls laid more than forty-five eggs for the
entire period, while none of the experimental birds laid more than
forty-five eggs during the experiment. One of the thyroprotein-fed
birds died during the experiment, and the records of this bird together
with the records of the corresponding paired bird in the control group,
were eliminated from the final compilation.

Following an average egg production of 16.9 percent during the ini-
tial period, the three year old controls went out of production complete-
1y during the next interval, and laid only one egg during each of the

next two intervals (Tables 12 and 13, and Figure no. 3). The experimntal

30

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

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34

birds laid at a rate of 22.1 percent during the first period, then drop-
ped.to 4.1 percent from May 26 to June 23. From.June 23 to August 18,
while the controls were laying only two eggs, the experimental birds
increased their average egg production to 19.4 percent on July 21, and
20.4 percent on.August 18. The therprotein—f d birds showed a seasonal
decline from.lugust 18 to October 13, dropping from 20.4 percent to 0.5
percent during this time. Although the controls aseraged 3.3 percent
during the last three intervals, only three-birds were laying during
this time. The average egg production of the experimental birds climbed
to lO.2 percent during the last period. The thyroprotein-fed birds mainr
tained a much higher egg production throughout the experiment except for
the interval of September 15 to October 13, and their overall average
was 11.1 percent, while that of the controls was 2.7 percent. As with
the two year old birds, one of the experimental birds died during the
experiment, and the records of this bird along with those of the corres-
ponding paired.bird in the control group were not included in the final
tabulation.
Two and Three Year Old Birds - Feed Consumption

The feed consumption of those birds that died during the experiment,
and the feed consumption.of their partners was included in the final
computation (Table 14). In addition the feed consumption of the two
and three year old controls was combined, as was that of the experimen-
tal two and three year olds. The thyroprotein-fed birds consumed
slightly more feed throughout the experiment than did the controls, the

difference ranging from 1.1 to 0.2 pounds per bird.

TABLE 14 - Feed Consumption

 

 

Feed consumption per bird by 4 week intervals

 

 

 

 

May 26 Jun 23 Jul 21 Aug 18 Sep 15 Oct 13
to to to to to to
Jun 23 Jul 21 Aug 18 Sep 15 Oct 13 Now 10
Lots III a v - Two?
and Three Year Old
Controls 6.1 6.0 6.1 6.0 5.5 5.8
Lets IV a VI - The
and Three Year Old
Experimental Birds 6.3 6.2 6.2 7.1 6.7 6.1

 

 

 

 

 

 

 

 

35

36

Discussion

Egg Production

The lewel of ten grams:of thyroprotein per one hundred pounds of
feed was found to be the most nearly optimum in the experiments conduct-
ed by Turner, etc. a1. (1945), however it would appear that this level
was not optimum in this particular experiment. This would indicate that
the Optimum level for different strains and breeds of birds would tend
to vary. Since the previously mentioned experiments were performed in
lissouri, the further possibility of the environment affecting the
optimum level exists. In addition the thyroprotein used in this experi~
sent tested 3.04 percent thyroxine by chemical assay, while that used
at Missouri tested 2.73 percent thyroxine by biological assay, which
would imply that there was a difference in the actual amount of thyro-
xine awailable.

Inasmuch as the results of this experiment indicate that the level
of thyroprotein, or at least the amount of available thyroxine, was not
optimum, the question arises, was the level too high or too low! The
egg production of both the experimental pullets and two year old birds
was consistently depressed in comparison with that of the control birds
of these two groups (Tables 1, 2, 10, and 11). However, the egg produc-
tion of the experimental three year old birds was decidedly stimulated
in comparison to that of the three year old controls (Tables 12 and 13).
Since thyroid activity is known to decrease with advancing age, Crew
(1925), there is a possibility that the optimum lewel for the three
year old birds was slightly lower than that of the two year olds or the

pullets. Due to the limited number of birds available only eight of

~JC.

3?

bath.the two and three year old birds could be employed in this experi-
ment, and this is a.rather small number from which to draw any definite
conclusions. Despite this the average egg production of the experimen-
tal three year olds was definitely greater, particularly during the
months of highest mean temperature, than that of the corresponding
controls.

The ten gram level of protamone (2.73 percent thyroxine by biologi-
cal assay) supplied the chicken with an amount of thyroxine in an excess
of that amount normally secreted by the body, Turner, Kempster, Hall,
and Reineke (1945). In the experiments of Turner, et. al. (1945), a
twenty gram level of thyroprotein was found to have a depressing effect
on egg production and was therefore considered in excess of the optimum
level. While the results of this experiment imply that there is a
possibility that the ten gram level was slightly below the optimum in
this particular case, the above information indicates, that the ten
gram level need be increased only a.few grams, before favorable results
might be effected.. Therefore, it follows, that this work might have
produced.more conclusive results, if levels of twelve and possibly
fifteen grams of thyroprotein had been included; however, a limited
number of birds prevented such procedure.

It is interesting to note that in each of the three different age
levels of birds employed there was a definite tendency on the part of
the experimental birds to resist the normal seasonal decline in egg
production during the period of July 21 to August 18, which was the
period of the highest monthly mean temperature. In addition the season-
al decline of the experimental pullets and the experimental two year

old birds, was somewhat retarded in comparison with that of the control

38

birds of these two age groups. Although these results can not be consid-
ered.decisive, there does exist some evidence that the feeding of thyro-
protein may yet be found to have value in increasing egg production by
delaying the normal seasonal decline. Apparently the feeding of thyro-
protein supplies the birds with a constant level of thyroxine throughout
the laying year, whereas high temperatures probably cause a reduction

in the normal rate of secretion of thyroxine by the thyroid gland, and
consequently the egg production undergoes a seasonal decline during the

suns-er and early fall.

Body Weight and Feed Consmption

The body weight of the control pullets was greater than that of the
experimental pullets (Tables 3 and 4). The greater weight of the con-
trols remained at a fairly constant rate throughout the experiment.
This would indicate that the controls were heavier as a group than the
thyroprotein-fed birds, and that the level of thyroprotein (available
thyroxine) employed here had no noticeable effect on body weight.

The control pullets consumed more feed during the experiment than
did the experimental pullets. In general, however, the feed consumption
of the controls was not markedly greater than that of the thyroprotein-
fed birds. Furthermore the controls maintained a higher average egg
production and were a heavier group of birds, and for these reasons
would naturally consume more feedL Therefore, it can be assumed that
the ten gram level of protamone used here, had no marked effect on feed
consumption.

The two and three year old thyroprotein-fed birds consumed slightly
more feed than did the controls of these two age levels (Table 14).

Since the average egg production of the two and three year old experimental

39

birds was slightly more than that of the corresponding controls, it
follows that their feed consumption would tend to be a little greater.
As a result this slight difference cannot apparently be attributed to

any influence of the iodinated casein.

Holt

The experimental pullets had.pr0gressed much further in their molt
than had the controls, when the first check was made (Table 5). How-
ever, the results obtained from the next two observations, indicate
that the two groups were melting at approximately the same rate. These
results would further imply that the thyroprotein-fed birds started
their molt at an earlier date than did the controls. The fact that the
controls began their molt at a.later date would account to some extent
for the superior average egg production of the thyroprotein-fed birds
during the last four week interval of the experiment. There is some
evidence here to indicate that the feeding of thyroprotein may hive had
some effect on the molt, since the experimental birds began their molt

at an earlier date than did the controls.

Egg Shall Quality and Egg Weights

The eggs laid by the control pullets had a slightly smaller rate of
moisture loss in grams than those laid by the experimental birds during
the time that the first egg shell quality test was conducted (Table 6).
This difference was not great enough to be significant, however. The
moisture loss rate of the eggs of the two groups used in the second egg
shell quality test differed even less than that of the first test. Com-
parison of the moisture loss rate of the first and second tests for egg

shell quality (Table 7), indicate no significant differences due to

40

seasonal variation. The negligible variations in moisture loss rate
obtained in this experiment indicate that the feeding of protuone at
the level employed had no effect on egg shell quality.

The differences in the egg weights of the two groups of pullets
followed a similar pattern throughout the experiment (Table 8). Here
again, the results did not differ sufficiently to infer that the level

of thyroprotein herein employed had any influence on the egg weights.

41'

Sumry

Three different age mups of Rhode Island Reds received a ten gran
level of thyroprotein (3.04 percent thyroxine by chemical assay) per
one hundred pounds of feed. The average production of the experimental
pullete was 29.1 percent, as against 37.? percent for the controls.

The two year old controls outlaid the experimental birds 20.1 percent
to 10.2 percent. Three year old experimental birds were apparently
stimulated to a production level of 9.0 percent above that of the controls.

The rate of decrease of the seasonal decline of the experimental
birds was somewhat retarded in comparison with that of the controls,
particularly during the interval of the highest temperature, July 21 to
August 18.

In two tests ads for egg shell quality, so significant differences
between the moisture loss rate of the two groups of pulleta were obtained.
In addition it was found that the level of thyroprotein, or at least the
amount of available thyroxine used here, had no noticeable effect on
body weight, mortality, egg weights, or feed consumption.

Although the two groups of pullete melted at about the same rate
from October 5 to November 9, 194?, the experimental birds had melted
about 12.4 percent more by October 5, than the controls had. This dif-
ference indicated that the feeding of thyroprotein apparently resulted
in the experimental birds going into the molt sooner.

The lack of agreement of these results with those of the work of
Turner, et. a1. (1945) indicates that results obtained in one experiment
may not hold for a similar experiment performed with different birds in

a different environment, or with different amounts of available

thyroxine. Furthermore, more work is necessary to establish the optimum
levels of thyroprotein (available thyroxine) necessary for different

strains, varieties, and breeds, and for different environments.

42

43

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