• TABLE OF CONTENTS
HIDE
 Title Page
 Acknowledgement
 Table of Contents
 List of Tables
 List of Figures
 Introduction
 Literature review
 Experimental procedure
 Results and discussion
 Summary
 Bibliography
 Appendix
 Biographical sketch














Group Title: effects of a vitamin A deficiency on the reproductive capacity of rams
Title: The effects of a vitamin A deficiency on the reproductive capacity of rams
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Permanent Link: http://ufdc.ufl.edu/UF00097944/00001
 Material Information
Title: The effects of a vitamin A deficiency on the reproductive capacity of rams
Physical Description: ix, 156 leaves : ill. ; 28 cm.
Language: English
Creator: Sosa, Enrique, 1940-
Copyright Date: 1964
 Subjects
Subject: Rams -- Feeding and feeds   ( lcsh )
Animal Science thesis Ph. D
Dissertations, Academic -- Animal Science -- UF
Genre: bibliography   ( marcgt )
non-fiction   ( marcgt )
 Notes
Statement of Responsibility: by Enrique Sosa.
Thesis: Thesis (Ph. D.)--University of Florida, 1964.
Bibliography: Bibliography: leaves 92-96.
Additional Physical Form: Also available on World Wide Web
General Note: Typescript.
General Note: Vita.
 Record Information
Bibliographic ID: UF00097944
Volume ID: VID00001
Source Institution: University of Florida
Holding Location: University of Florida
Rights Management: All rights reserved by the source institution and holding location.
Resource Identifier: alephbibnum - 000406503
oclc - 24680729
notis - ACF2774

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Table of Contents
    Title Page
        Page i
    Acknowledgement
        Page ii
    Table of Contents
        Page iii
    List of Tables
        Page iv
        Page v
        Page vi
        Page vii
    List of Figures
        Page viii
        Page ix
    Introduction
        Page 1
        Page 2
    Literature review
        Page 3
        Page 4
        Page 5
        Page 6
        Page 7
        Page 8
        Page 9
        Page 10
        Page 11
        Page 12
        Page 13
        Page 14
        Page 15
        Page 16
        Page 17
        Page 18
    Experimental procedure
        Page 19
        Page 20
        Page 21
        Page 22
        Page 23
        Page 24
        Page 25
        Page 26
        Page 27
        Page 28
    Results and discussion
        Page 29
        Page 30
        Page 31
        Page 32
        Page 33
        Page 34
        Page 35
        Page 36
        Page 37
        Page 38
        Page 39
        Page 40
        Page 41
        Page 42
        Page 43
        Page 44
        Page 45
        Page 46
        Page 47
        Page 48
        Page 49
        Page 50
        Page 51
        Page 52
        Page 53
        Page 54
        Page 55
        Page 56
        Page 57
        Page 58
        Page 59
        Page 60
        Page 61
        Page 62
        Page 63
        Page 64
        Page 65
        Page 66
        Page 67
        Page 68
        Page 69
        Page 70
        Page 71
        Page 72
        Page 73
        Page 74
        Page 75
        Page 76
        Page 77
        Page 78
        Page 79
        Page 80
        Page 81
        Page 82
        Page 83
        Page 84
        Page 85
        Page 86
        Page 87
        Page 88
    Summary
        Page 89
        Page 90
        Page 91
    Bibliography
        Page 92
        Page 93
        Page 94
        Page 95
        Page 96
    Appendix
        Page 97
        Page 98
        Page 99
        Page 100
        Page 101
        Page 102
        Page 103
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        Page 119
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        Page 121
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        Page 129
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        Page 131
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        Page 136
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        Page 150
        Page 151
        Page 152
        Page 153
        Page 154
        Page 155
        Page 156
    Biographical sketch
        Page 157
        Page 158
        Page 159
Full Text












THE EFFECTS OF A VITAMIN A DEFICIENCY

ON THE REPRODUCTIVE CAPACITY OF RAMS














By
ENRIQUE SOSA


A DISSERTATION PRESENTED TO THE GRADUATE COUNCIL OF
THE UNIVERSITY OF FLORIDA
IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE
DEGREE OF DOCTOR OF PHILOSOPHY


UNIVERSITY OF FLORIDA
August, 1964














ACKNOWLEDGMENT


The writer wishes to express his sincere appreciation to

Dr. A. C. Warnick, Chairman of his Graduate Supervisory Committee and

Dr. T. J. Cunha and Mr. R. E. Deese under whose guidance and supervision

this study was made.

Grateful acknowledgment Is extended to Dr. R. L. Shirley,

Mr. P. E. Loggins, Dr. H. D. Wallace, Dr. F. E. Neal and Dr. W. P.

Callahan for their efforts and counsel extended freely throughout this

study.

The writer expresses his appreciation to Mr. R. C. Dees, Herdsman,

for his assistance in the feeding and handling of the experimental ani-

mals.

The assistance of his fellow graduate students, Mr. J. Easley,

Mr. C. Piedra and other personnel at the Nutrition Laboratory is also

appreciated.

The advise of Dr. A. E. Brandt in the statistical analysis is

gratefully acknowledged.

The writer is indebted to his wife Irene for her patience and en-

couragement during his graduate study.


















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31 Analysis of Variance Mean Squares for Semen Volume,
Motility, Abnormal Cells and Total Sperm Cell
Production--Week 2 . . . . . . .... 144

32 Analysis of Variance Mean Squares for Semen Volume,
Motility, Abnormal Cells, and Total Sperm Cell
Production--Week 3 . . . . . . .... 145

33 Analysis of Variance Mean Squares for Semen Volume,
Motility, Abnormal Cells, and Total Sperm Cell
Production--Week 4 . . . . . . . 146

34 Analysis of Variance Mean Squares for Semen Volume,
Motility, Abnormal Cells, and Total Sperm Cell
Production--Week 5 . . . . . .. . 147

35 Analysis of Variance Mean Squares for Semen Volume,
Motility, Abnormal Cells, and Total Sperm Cell
Production--Week 6 .. .. .. .... ... .. 148

36 Analysis of Variance Mean Squares for Semen Volume,
Motility, Abnormal Cells, and Total Sperm Cell
Production--Week 7 . . . . . . . 149

37 Analysis of Variance Mean Squares for Semen Volume,
Motility, Abnormal Cells, and Total Sperm Cell
Production--Week 8 ................ 150

38 Analysis of Variance Mean Squares for Semen Volume,
Motility, Abnormal Cells, and Total Sperm Cell
Production--Week 9. . . . . . . .. .151

39 Analysis of Variance Mean Squares for Semen Volume,
Motility, Abnormal Cells, and Total Sperm Cell
Production--Week 10 . . . ... ..... 152

40 Analysis of Variance for the Difference in Mean
Size of the Testes . . . ... . . . 153

41 Analysis of Variance for the Difference in Mean
Size of the Epidydlmis . . . . . . . 153

42 Analysis of Variance for the Difference in Mean
Size of the Adrenal Glands . . . . .... 154

43 Analysis of Variance for the Difference in Mean
Size of the Seminal Vesicles . . . .... 154

44 Analysis of Variance for the Difference In Mean
Size of the Cowper's Glands .. .. .. .. .. 155











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LIST OF FIGURES


Figures Page

I Cloudy Condition of the Cornea Present in a
Vitamin A Deficient Ram . .. . . . . 30

2 A Vitamin A Deficient Ram Showing Extreme
Weakness . . . . . . . . . . 30

3 Effect of Vitamin A Supplementation on Body
Weight Changes of Rams .. . . . . . . 43

4 Effect of Various Hormone Treatments on Body
Weight Changes of Vitamin A Deficient Rams . . 44

5 Effect of Irradiation on Body Weight Changes
of Vitamin A Deficient and Normal Rams . . .. 45

6 Effect of Vitamin A Supplementation on Mean
Semen Volume of Rams . .. .. .. .. ... 54

7 Effect of Various Hormone Treatments on Mean
Semen Volume of Vitamin A Deficient Rams .... 55

8 Effect of Irradiation on Mean Semen Volume of
Vitamin A Deficient and Normal Rams . . ... 56

9 Effect of Vitamin A Supplementation on Mean
Sperm Motility of Rams ............. 59

10 Effect of Various Hormone Treatments on Mean
Sperm Motility of Vitamin A Deficient Rams . . 61

11 Effect of Irradiation on Mean Sperm Motllity of
Vitamin A Deficient and Normal Rams . . ... 63

12 Effect of Vitamin A Supplementation on Per Cent
Abnormal Sperm of Rams. . . . . . .. 65

13 Effect of Various Hormone Treatments on Per Cent
Abnormal Sperm of Vitamin A Deficient Rams . . 67

14 Effect of Irradiation on Per Cent Abnormal Sperm
of Vitamin A Deficient and Normal Rams . . .. 69














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INTRODUCTION


Efficient reproduction in his animals is one of the goals that

any producer must achieve in order to successfully compete in today

livestock business. This underlines the importance of research in any

aspect of reproductive physiology.

In the literature one finds that efforts in this area of research

have been directed mostly towards studying various effects of environmental

and genetic factors on female reproduction, somewhat underestimating the

importance of the male; consequently the basic aspects of the male repro-

ductive physiology have been overlooked to some extent in the past.

This study Is part of a broad project being undertaken at the Uni-

versity of Florida to Investigate the effects of various nutritional fac-

tors on the male reproductive capacity. Other studies in this project

have been the work reported by Meacham (1962) and Tilton (1962) in which

protein and energy have been investigated.

Lack of vitamin A is the only specific dietary deficiency that has

been shown with any certainty to affect semen production in farm animals.

It is also true however that the studies from which this knowledge was

obtained are to some extent outdated in the light of today improved re-

search methods and techniques. The use of purified diets is indeed a tool

of great value in animal research for It facilitates complete knowledge

of the nutrients present in the diet, thus allowing a more precise charac-

terization of the role that a specific dietary factor has in the physiology











of the organism in question. It Is with this in mind that t-*- p ,reer

study was undertaken to further Investigate tr'e ii'fcril of a,,raci.r..r,.

A on the reproductive capacity of the ram.

Due to the ever increasing need of information related to rjdrlal:--.

and its effects on biological systems, a study on the tolerance of ., a.Tii.

A deprived animals to gamma irradiation from a Co source was inclJdEJ.

Also attempt was made in this study to counteract the harmful

effects of vitamin A deficiency by administering Testosterone, preg-..a..

mare serum and Thyroprotein.














LITERATURE REVIEW


Vitamin A and Reproduction in the Male

As early as 1924 it was found in experiments with rats by Gross (1924)

and Wolbach and Howe (1925) that vitamin A deficiency caused pro-

nounced atrophy of the testes. Since then extensive investigations have

been carried out to elucidate the influence of this deficiency on the

male germinal epithelium in laboratory and farm animals. In addition to

atrophy of the testes this deficiency will also cause impaired develop-

ment of the epididymis, seminal vesicles, and prostate in young rats.

Injection of testosterone can obviate this atrophy in vitamin A deficient

rats as shown by Mayer and Truant (1949). Further injections of gonado-

trophic hormone will stimulate the development of the accessory sex glands

In vitamin A deficient rats according to Mayer and Goddard (1951). This

suggests that this deficiency causes a lowered excretion of pitituary gona-

dotrophic hormone or a lowered response to this hormone by the interstitial

cells of the testes.

Recent work by Howell et L. (1963) showed that male rats main-

tained on a diet in which the vitamin A alcohol had been replaced by

vitamin A acid developed lesions in the reproductive tract. The testicular

changes were a sloughing of the cells of the germinal epithelium followed

by an obliteration of the lumen of the tubule by sertoll cells. Testicular

regeneration was produced by the administration of vitamin A alcohol. The

lesions were comparable to those of vitamin A deficiency as described by












aio.- (9I I: tuL- ir*CF unciOCpLilC3ted bt the i conddr .ani bnfetati ;cn. Of

u t rir.I h d ielc ienc Tr.i paper I t r-e r lr t I0 dc cr .e le I r.s .n

Ih. tIc : IC heal in., )rOic ..i' j pra pubtFrt I r it fe. (., jit 3a.ln A acid

diet It I1 kid I cccipted Etat vr.ti -Tin A acid car. perfor.c. all .ce

Su-ci c 1C.... c r i A Iclil-n l -; CcePpf ior tT part 1, 1 .. .,. DI r l.1o 9

anrd wald li9c0l :ujuar iz; d arie rrle or vlwiArin A acid in a recer.r a,.T|-

p.:..16 i . itI a-Ir A. A Tr.e general i, uc ru-.cilur. .:.i li.litr.r, A that iup-

port growth an.d c.iratr,l.5c.i c n thee rat are itrafd dl ;.:. 1 l.a.nin A .30.d.

bul ;ACte t 6c E a EduC hi. t rd i t. :u I .anc t i or. r. ti t [er ite

Sicor.oi ir., form, n .,r.r i. l[ T A i t.frcd. *..:.r ir'e sidc, ide retirer ere)

r. edc ,,:,r (h: ,,-i I | ,. Url i ud I p: t ,.' .i ; r 1 i A . .: r : ; i I 3 i .r -




c:.i .-. ., Lni. e r i r.i t LI Ctuol i nd roeIn f allure




luter tcr':En .r ihe I ; ual c I det er ior are. r, i.:.l d ry the ir: ; u

a' ll L a cr a i. :1 trc ;ci r. t .cil: i t ir. 3r. ti he ,ri c c pcarir.d ret in .

Tie r..din j ..,f M. h ii et i. (l L I ., c t.-.:i zuFpor th e iata c 't'i oi

r[' *.j u i l. IA l tI W. I i,. I i

In t- i i: a prcn.our.c.J .'J..i rC dciC icc,' dec a : i -uaf *a...u-

r ty ; upprew e. .c ; ua' i.rt-ric:t a.t Cju:c (aSi i Iular deene r iu ic.n rij

thu a I&.er rg c.f :pert0 pFr duct u C nm Qut .In, Ir I has I:e, nhon, r ,r.

Irt att r. et ai (i ..i t Erbt et a (i 4.) a.iJ tL i.t-Ert arij M.art I i 35).

b atg .n [ a i 1 1 .S1 studied the af riec of r nation |.:.. .r caruter.e or.

ihe hreedji.-j ber3,ior 5 eraI. t:.ye.-c s: : an-id Ee.Tr. prd.c ior. of *-.atu'e

Jair, bulli. Laec I o 2 1to "cy o* .i ra,.i,. A pE r i0i .TI. .:.f bl,:.yj

nere recoridcd th-f.:.re ar., ut the Jef icienc, i,.7>p e. appeared










Incoordination and loss of the ability to mount without loss of libido

were the first signs to appear and they occurred before any marked impair-

ment on semen quality was evident. Percentage of motile spermatozoa was

decreased. The per cent of abnormal spermatozoa was increased. Histolo-

gical examination of the testes showed degeneration of the germinal epi-

thelium of the seminiferous tubules. Hogdson et l. (1946) also found

that vitamin A deficiency apparently does not cause severe disturbance

in spermatogenesis until the animals are in advanced stages of deficiency.

Erb e al. (1947), Hodgson et al. (1946), Jones etal. (1946)

and Madsen etal. (1942) have reported that pituitaries are often cystic

In the vitamin A deficient bovine. These findings suggested the possi-

bility that vitamin A deficiency may exert its deleterious effect on re-

production indirectly through the anterior pituitary gland. Gunnet al.

(1942) improved the semen quality of a ram deficient in vitamin A by

injections of pregnant mare serum. Hogdson et al. (1946) obtained similar

results with a vitamin A deficient bull. However Lindley et al. (1949)

reported that testosterone propionate injected semiweekly in dosages of

12.5 mg. before and during the deficient condition or of 37.5 mg. after

vitamin A deficiency symptoms began to appear resulted in no improvement

on semen quality of vitamin A deficient rams. Similar results were ob-

tained from Injections of pregnant mare serum. This work suggests dif-

ferences between rat and sheep since Mayer and Truant (1949) were success-

ful in preventing testicular atrophy in the rat resulting from a vitamin

A deficiency by administration of testosterone propionate.











Pathology of Vitamin A Deficiency

Moore (1960) described at least three basic lesions occurring in

avitamlnosis A: (1) Lack of vitamin A, required in the form of its alde-

hyde for the formation of rhodopsin, causes defective dark adaptation

unless complications have ensued this lesion may be regarded as "bioche-

mical" rather than "structural." Thus the response to treatment with

vitamin A is rapid and dramatic. (2) Lack of vitamin A causes xerosis

or keratinization of membranes in many parts of the body, the well-known

xerophthalmia belongs to this group of lesions. Epithelia tends to

become dry and excessively thick and horny. Those membranes having a

columnar structure, often associated with secretions of mucus give place

to thick layers of stratified epithelia as found in the outerpart of the

epidermis. The most common secondary effects of avitaminosis A arise

from the bacterial infection of the abnormal membranes. In some sites

such as the urinary bladder, the injury to the membrane may sometimes

induce calcification with the formation of stones. (3) During growth,

lack of vitamin A can cause defective modeling of the bones. As a result

the bones are not compact, strong and well shaped, but cancellous, weak

and excessively thick.

Lesions of more than a single type are often super-imposed, and

the possibility of further types cannot always be regarded as secondary

effects of the bone lesions. Another abnormality Is an increase in the

pressure of spinal fluid sometimes associated with hydrocephalus. These

lesions might well be regarded as secondary effects of malformation of

the skull bones, but a rival theory suggests that deficiency of the vitamin

causes the chloroid plexus to secrete unduly large amounts of fluid.











Vitamin A generall)

Vitamin A is an alcohol and forms esters with fatty acids. Most

of the vitamin A in the livers of both mammals and fish is present in

the sterified form (White et al. 1959 and Harper 1963). Conversion of

provitamin A (carotene) takes place in the Intestinal wall in rats, pigs,

goats, rabbits, chickens and sheep. According to Barnett and Reid (1961)

carotene may be found in the liver of animals suffering from a vitamin A

deficiency and does not appear to be capable of utilization for vitamin

A synthesis. Dowiing (1960) summarized today'sviews on vitamin A metabo-

lism as follows: Vitamin A alcohol, the transport form of the vitamin

and the storage form as the ester, is oxidized to retinene because of the

rapid removal of the aldehyde in its combination with opsin to form rho-

dopsin. The alcohol is oxidized to the acid in the liver and it is rapidly

used in its function of growth and tissue maintenance. Acid is the im-

portant form for all functions other than visual, it must be stored as

the alcohol and cannot be reconverted to either aldehyde or alcohol.

Most of the information on the concentration of vitamin A in the

various organs has been gained in studies on rats. Moore (1931) used

the antimony trichloride reaction, with the aid of a Lovibond Tintometer,

In studies of vitamin A in the tissues of rats which had been given liberal

amounts of carotene. By this treatment total stores of up to 50,000 I.U.

of vitamin A were accumulated in the liver. Apart from the intestine

which contained much yellow pigment the rest of the carcass contributed

100 I.U. of vitamin A. When the various tissues were examined separately

the intraperitoneal fat contained 0.5, 5 and 5 I.U. in four experiments,

and the kidneys and lungs each 0,0 and 5 I.U. in three experiments.













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[i.: rc..,3j(,A ir.9 [rre u. 5 i ** ;," I ai r.' A ;r, Iarg:e *.- n C

d'l., :e .C .C-ic r[ .r.- :.f ,'JLL' I U.. pa"i qp r s.. Ii.cr, 3JU I .1. pC( 'ara'

kidr: ,j 2":1.' i U per ,ra.. ,:.r I n., ,ifrc >:,bt in d. TJr., Ir. rAperit.:r.cl

I t 3C p t. i r d '.h I .I. pc r grj. >.J Che :drena I anrd I II 1. p.e r

ra.T. 8, r i I- .4 I h Jul. I ', i-e ac reiC .C rt" I er Cui. c :. ;ild uD t.

.,C :" I.U. per .r ,.a.. rhc I d',,,.: ,c l.U. l.,-, E.: ,.'I I .U. And :u, pr.i -

re ,-ale: -> Il.U. C.r.cer.r- c. Ci uP to 1 . per rr t ar ere rt:.und Ir.

Stihe i. C 1: Tr art :P ;r. i-,j t r i..

Tr.. (,: al : (urag- Cf Lar.:E.-t and ta..,., 'n ., t rIe li r And de -

p:- I at jl C.:.: ,hit.| Iad 02 -Cc: Ch: C r: b L I .- rC C rt6[ i .1 tclircuyr C. u

I .A: : I ...ated i :. E J. I:L I r -.i. f .i -. C A f or I r.e yC.un er

ar.i.:jl aiid up t' 3 q rj3- ;,'. jycJ Ci:, -*L M rird and Li-c4ll 19621. Fror.

r, t j C) pEr ce,-.i f the .tor aje di .r. (r lir I1 [--- :1r.j aP i ._.-:i

.:. i th, ..raqt ..A' I~r he fcr r.- oi tie- .. = 1.-. i el fr Cit i; 11-. tat

.ip.-:. car:t e .e pre .,r.itn tej. Tr,; :.rk r-porricd .:i Janre b; Gullib ri

iend Hart 119i) An 19;51. E At.:n llE1i) Iii .hown (riat I darry

cal 51 v Aor..r i.atel f 'c r r.,. r,i: fi aI e fc d a dE IeL ..n fs Ic r nc. dc -oiJ -3

I ain in C t I*t iE LlA E pl v .T-min I I; .cIl Jccrca r ir. a I;rn ar

*.Jmn.iL r u,..IiI a I lu3 e of rc. per Li ri are r- :.Chcd. C I .e: c rh bi q











such low plasma values for two consecutive weeks were shown to be depleted

of liver stores of vitamin A. Unpublished data by Dr. R. L. Shirley and

co-workers showed that cattle receiving 30,000 I.U. per day and those

receiving no supplemental vitamin A had blood plasma levels of 54 and 34

respectively. Another trial conducted during the summer showed levels

of 46 and 18 mcg. of vitamin A per 100 ml. of plasma for groups receiving

the supplemental vitamin A and no vitamin A respectively. Vitamin A in

liver expressed as mcg. per gram of liver dry weight was found to be 119

and 78 for the supplemented group during the first and second trial res-

pectively and 17 and 7 for the second group during the first and second

trial respectively. In the second trial vitamin A level in the heart was

found to be 0.17 and .05 mcg. per gram of heart for the supplemented and

non-supplemented groups respectively.

In sheep vitamin A storage has not been as extensively studied

as it has been in rats. Harm (1942) reported the average vitamin A con-

tent in a gram of liver as 503 l.U. in 19 animals. Moore ad Payne (1942) reported

levels of 460 I.U. per gram of liver in 20 animals. The above data were

expressed in terms of grams of dry liver. Recent data in the literature

indicate the levels of vitamin A in the liver of sheep to be within a

range from 25 to 54 mcg. per gram of fresh liver, Cline (1962), Goodrich

(1962) and Cline et l. (1963). Data on the vitamin A content of tissues

other than blood and liver are lacking for sheep.

The Thyroid and its Role in Reproduction
and Vitamin A MetabolIsm

The role of thyroxine in reproduction has been most extensively

studied among the hormones that may directly or indirectly influence the











secretion rate of hormones influencing repr jj.ucC ac I. i t I :utci a: iLh

gonadotrophs and the hormces of the ovary an'd ic .ti.

Berliner and Warbrltton (1937) reported trat sth/irtodct:.., oi

rams produced a decrease in semen volume, scerr....alz:..,-, c..r.'fl
an increase in the relative numbers of abncrreai :pr... r..,r ., 'n ad..i-

nistered to the thyroidectomized animal re!ulrEa .r. tic :r.A .uci; -. cf

semen with an increased number of sperm and djcra:ced perier.r le .:.r jb,,.r-

mal spermatozoa. Bogart and Mayer (1946) rp..:rted tii.a trEac.c.ient ..lt.

thyroxine and thyroprotein alleviated sympr.iE-: 1 .j,-Tr.er tie' ll, re',il-

ting from impaired spermatogenic activity. Tr,e reeJi.g r Lt;.r.u.a i

during the fall breeding season maintained ,.-h'e.. tjractCeris [.:; t,i~iCl

of the summer months. It was concluded in tr,,-i :ud, I[r. the E[.,r:.,d

gland is of major importance in the reproduciii e 1 h, ,i.:.3, :r Er_, ra,.

Bhatnagar (1955) studied the seasonal variar ..i-. in th, rist:log, ,r te

thyroid and the testes of Indian buffalo bull- TI.- tiouL.J :..,l eir ,jL E

seasonal variations in the histological str. ucture (r.e (.:. jlai.n.

they also observed that during the period thenr the ,ual., ..,' ie.Tn jeri-

riorates in buffalo the secretary activities: ..4i r.e t .. jl j..d: j rh,..ar.

by their histological studies appears to decre.:. GC:-,.. %1 i 2 ,-

ducted an experiment to study the effect of tr.,rc.,n: and cr- -,ant ..are

serum (PMS) hormones on reaction time and .i r,-. qualir, :.r tui,i5 bl ll:.

The experiment was conducted in India from Ju-,r. ij Febr-t,, One tru 5-3..j

I.U. of PMS were administered by weekly subcutla. IIeu: Irnjecr *..: n

daily dosage of 100 mg. of thyroxine was ac..,;nisterid orallr Trca. t.e,-,

with thyroxine and PMS did not influence the reaction t'.,, .:.r t, ar.I,,,aTi.














: r I :.: l r : L lh, r ri.r,- r, .












i., thyroprotein in rams. Ten mg, of testosterone and 1 gm. thyroprotein

.-:re .Jd.... ri red daily by subcutaneous Injections and orally respectively.

O.ne ,.. of thyroprotein per hundred lbs. of body weight was given orally

t:* a second group. A third group received both testosterone and thyropro-

ten at the same level administered to groups one and two. Thyroprotein

reduced weight, decreased libido and had no effect on sperm motility,

.iume, concentration or number of ejaculates. The testosterone treated

gr:;up showed a significant decrease in semen concentration. They also

repr.rr. that testosterone could prevent the loss of libido caused by

crain.ern. with thyroprotein.


Recent studies by Brooks (1961), do not support the obser-

vations of Berliner and Warbritton (1937), and Bogart (1946) Brooks

(1961) injected thyroxine at levels of 2, 3, or 4 mg. per hundred Ibs.

of body weight to rams. Considering the fact that 1 mg. of thyroxine

injected equals approximately I gm. of thyroprotein fed (Cole and Cupps

1959) the dosages used in the experiments of Brooks (1961) are compar-

able to dosagA% used in previous work in this field. The results

obtained by Brooks (1961) indicated that injections of












Er,r:. rair.c t hi)h [e.T-.pcr6 Eulc w C.Ec.-.E[u ittd JI Ic d1eE r i nT r i I er'rir : r,.,

-e ] r I 'i L . I apa :.l ..a: l., iEt .l 3'.- r ue pr c.*uuCd

a h,p r .ld C., -.11 j". but t.li t i j r... r f'e : .:.r. i,-, .. Ha,. e lcr ..-.e

r.uit L C C.u luu -....dal *. , y.46-eI l Iz tiO at hiT h .....I ;r'tCE [hE

S rl .:. aI r rep:r [ .E .'.i j ,-r Ja-'iJer i de r -.je .j1 ... r .- .r,[ 1 :. ,, i -

(i.:.r .ih r, ..:.. i ar.,ut,(E.I h .. ei- :( r. t1-' cl,rf, a3 C cidl 0'

the. ? .ri..aIl:. .utC. u, r..:r- a: Tur. n r \Ilcl'. LI'. a.d Lupp 1 J591 a..J

MaleZ (IrI l I .) r..? t ai [i It., ui pl 1.c . ;.-..I d j r. itr. repr.,:ucti.-

i: r .e: tut i. i p. .rI, u.-dc r [ *.-- B N. ,,

,-.i[i lc i:.:.-,rI u'.de r:[ c r.-.c- ti.r, .:.r I t. L r .,r, 3 ,d E-i j[ .

re li[ *. -,h;p I i [ ,,,-, ..E b. ,He. LrI, ( i 4. I re .i e.- d [ : I- 5 :, ,-

j C( rC ..d r 5C ed i e rai .. 1 v F ;I<.o I ,: j I I The lait .L f [ ...i,, A I r.-

asc ihr i.c i. i at- d I.,p r r...T..ir.:. r .. i [t.t opp:: ct h t C .

1 I r l ;C r.:, 1i1- t L-r E' i 11 pir J .- r. p rc r opr., r the [ t e : .j

I ;1 F c r c I 3.,-, rl t,p rt.,r :.i 1 r;s. ir, rcr .'-: e ui .-.,i : r : r O it i,- A .

14I I -. (1-. Ercc t, t.-,r r .d Car.I.-.cE c. C.u l LE U .... c iei [., ..[ Et .. r

v ; t theic ..-"E .-: r > i r. ha tb.v c.:..-.r r.- d E j rk .j..e :...c

Ir i I I I ;-' ,' ,c3 : rC'. I.. -. h :,r ,-,% vc. .l j b u r.a.-l i i, u C 1 [i.ar 1,J t r

i.,rc..- .c .-cre : *.: pe., t ur .:r, ji r .t da ,-, d r.r r ar c...ever

hc p .-.i. .Su(t t.r.i [t1-6 r r culti ru C).-ipl ; : ic l tb acC I.r trc .d r.Si.th

L.t:er...:d Ei n t. [h ., r.:, i,.. r c ratI ;rn,. hi , .i uI,,, ra. i r.:v CrEa: iC.

[I.E '. T.5 j ,-.i: ra. rutr'-c'.ti i.Ii:I d.j i t. .n I. R J a-.d TrI .3]-


(l ti l -..ri *'.; ..i h cai-ac rat ,t, I i-J L,. .,I rEp.: rIt 1 [.tr.a I. h ,p:,[, h

r1.5 : C.:.ri ;.,r of cJ it. ia la.T.i, A S- jr.C'J. c.d but *u -C.aIiI':dr ;r.

pI r C r. ,rt .idi :*T t. Ei r i.c r tu'I.r-t .l -.u.7.Ler ir ai-.,ITaI i r.c E I ;.ri.i'J..











Cline et al. (1963) found that feeding 150 mg. of tapazole per day or 100

mcg. of triiodo-l-thyronlne did not affect liver storage of vitamin A.

Any conclusion drawn from work where histological examination of the thyroid

glands were not made to confirm thyroidal status should be made with

caution.

Moore (1957) in his book of vitamin A reviewed the interrelation-

ships of thyroid and vitamin A metabolism. He concluded at the time

that the usual effect of thyroxine will be to accelerate any prevailing

trend In vitamin A metabolism. If conditions are conductive to expendi-

tures the rate of expenditure Is increased. If dietary conditions are

conductive to the storage of vitamin A the amounts stored will be increased

by thyroxine, however the why and how of this remains unsolved today.


Histology

Warbritten and McKenzie (1937) described the pituitary glands of

ewes in various phases of reproduction. They found the pituitary body

of the ewe to be very complex. They were unable to classify the cells

of the pituitary into the conventional acidophils, and chromophobes.

They found 9 morphologically different types of cells which they didn't

think necessarily represented distinct functional types. In this study

the different regions of the pituitary were found to be conspicuously

different, that counts made at one level would not be valid for another

or for the whole gland. At the time of the studies by Warbritton and

McKenzie (1937) present refined histological techniques were not avail-

able.













i-.i :. t cE cr r.i a r.ji .jre ..l Ir. .- t.:rct:,r if Cr, dui( : L F it he

L..r I: pl au ary ll. :. aric ( ~I5,': E n, w ,,I .. `n.i Ear.-. i1 35 '1 td.

r .p.: *d di Ec r e. .; i : l r. ,I j Kr., ar. rni r F r., :I Trhe, peri dic

?Cid .cr-.r i t c i. u E f A l: :f 1:- r : F I fl'jI .a- *., ed b, Jb.,ut nd FcEn.-e1

I I F, I tc .L t ., f 'jh I-c. ,.:, r Tp. .tc T, r r t I: I : CI c n : ,i '

a .-rul t':rC i-n..r, ce ll : r it Ic b A I. Br a nrd F. F I 3 E Ope ,n

-r I.p i r. F p : I: n ai T..r pr.,i, c nd unrc t iur i d c r-r'.Ce rft Fa

f.1r I ..cr Sc r i ted : r p.:.: r t ne r .r.ula It d jn-.J wi.t

lI e tr.il Ct pi ., M .:-. I r. Th.,r r r .-,r: C:r.l rre ('red : t ', ,rd l r

sI :c : 1 CI c itcreJ in t r. C:.r c* .4, rhe a. r '.C :.r i:.. The r uf ncr.j i

r Ilt i :..- :r-. 1 r t I.:e t 6 Cc l: id:C ii, Cr.C. i,[ h r.,r ., tr.:ci c h rT.rc-C .nc

pr-ducC F I :... T i:. : i F: 1 ,r eFr.1F r d icyr r.. IF SEd Si cu- .> trJ ..

Ih r..ud dcF i ic ''C T b. LIc t [ : I : crc iTrC I. l r I I r C r .idcd nail ih.rpi

-*u l *i* let" j. :-r r I tu c' a *icf rc r r 'u ar i Ei ; And r pe

Tt.e C- F I: I I c. Cr.. :e. -.t I pjrt C tFhe dCe ..)F- .5I5p C .h C.F C:., f -Ccr,-

Sr aicE j i . .ecui : 1 L5I.. .-Eritt or iJ bc. Thc.: E I r ec identi Il i iblc

.I Ch y;.rISad r : p. h iC aCt .I Ezp C i FI L T.. ere u- L lc tSL .. i Id-

tI r., c i6.ia 1 Pur..: 5s.-id CrI.. t-cr. (9i 5.. I 1I Jq Cr itCf .cd t c i.ate ..r.:

C.f : or.- i d-t-Irop t r e I ia c l I. tfhe r u -'.4 rec Po'. w' i-.OC r.. E Pe

I he, tI urI re : C(edCe C Cl p r Fi Ir .. re icti.r: ran r r-c cif.cr C.: .t r 1a S rtaF

t he- g9i,.ir. ir. addi tr, E CC r. r,djr.. rhoa. dirFr.;l Ic JF ti r i but l.nr.

r-.cE r.0.' :-atcd rhA the h c ;icph .ras .Crn..J.CrcfpFi C.. .5 c.i. ar ar* uira-

F C. arcd yF .. .7-T,rE .I.erI.t reSC li.o .. -ir. Pi. rh h.: -L F c lv c

nCtur rriri .' C-c': L.' I C geb: Ct f ," d l .r.phr CtC[c Ft CE[C lCer,.r, trcSalenr

ind aCf tr ir r. pEr i cD-I C f 0>-.adeCtC..M, E c .e .:r krr S uyc Ied CI8C











their "peripheral" gonadotrophs produced FSH and their "central" gona-

dotrophs produced LH.

Wilson and Ezrln (1954) described a modified PAS method in which

the Schiff reaction is followed by a methyl blue counterstain by which

some but not all of the PAS reactive cells changed from red color to

purple color. These workers concluded that the "PAS-red" cell represented

the Beta cell or thyrotroph and the "PAS-purple" cell the Delta cell or

gonadotroph. Rennels (1957) used a modification of the PAS-methyl blue

method which in conjunction with aldehyde fuchsin, enables'the tinctorial

differentiation of two types of gonadotrophs in the pituitary gland of

the rat. These two cell types were both gonadotrophic in function as

indicated by their development into distinct PAS-red and PAS-purple cas-

tration cells following gonadectomy. Thyroidectomy cells were found to

be PAS-negative, aldehyde fuchsin-negative and to have an affinity for

orange G. Sanders and Rennels (1959) found the predominant chromophilic

cell type In grafts producing luteotrophin (LTH) to be an elongated acido-

phil staining selectively with orange G when the azan stain is used.

Pearse (1960), Humason (1962) and Ham and Leeson (1961) were also used

as references for proper histological techniques, in this dissertation.


Effects of Irradiation on Reproduction

Schubert and Lapp (1958) described the spermatogonia as the most

radiosensitive cell in the human body and indicated that 50 roentgens

inhibits their development, permanent sterility for the human male re-

quires about 500 to 600 roentgens while 250 roentgens may produce steri-

lity for one to two years. Seminal fluid production was not affected











tu -n* I .4 .rr .-r; t. r c i T t rr. Ci E l I .-a e pr., d jd:.i* de-



CFj ..; F- I 41 C.C. Kti r-c uI rec.. .* ft 1 1 r r . ti rr E ., .n (lit

fertility of the male r i an.Jd mouse by t '[.ic ;.: c l;- matings 01' tr.e

treated animals. In both species irradiation with 300 roentgens p.i:-duce

a short period of sterility associated with oligospermia. This occurred

about 45 days after treatment in the mouse and 65 days in the rat.

Histological examination showed that the main effect is on the sper-.atL:-

gonial stages but there are obvious differences in the timing and e.ctr-.

of these irradiation effects. The time from treatment to the onset iC.

sterility is a measure of the overall duration of spermatogenesis. Tri-

highest dose level used 500 roentgens did not cause a rapid onset :,i

sterility in contrast to the action of some alkylating agents, whicir r4-

pidly induce infertility. in the cock X-ray doses of up to 10,000 r.-n-

gens has been shown by Kosin (1944) to have no detectable effect o- .*r-.

motility but the fertilizing capacity of spermatozoa was markedly rijlus-:t

after the exposure to 200 -i.enlqtge and was destroyed altogether :r'.tr

a dose of 500, supposedly due to damaged chr, -sai.;. Mann (1954) f..u.d

that rams exposed to luu.0'j0 r,:.t.-rr. and examined immediately af:.r C.-

posure had normal motil :.perT.altz.:. in their semen, fructolysis a-d a -

nosine triphosphate c.c..:-' [ ,cre :1:: normal. This would indicate ri.at

,I an damage -a: 3.:-r, Ir, l-.. ram semen it must be ascertained thrc.ugi.

ucr.,.r cr;terl. hM.r; i-re ad i- rish (1956) reported that when mature t.be

.ull *.-r.- p'P-J [* ; a ingr.ji- .ose of 100-400 roentgens whole-bod-, ga..aa

.'r.r.diari.on, Er, I rlru :.t.:, r ed s.',,-n changes were an increase in p.r

;nr. stIb ,r.Tsl _IPr iT[ .- I c,c after irradiation and a decrease i., ipcri.












.:-ne nt ra.:( --.r- alt *.gh.[ iEt ar Ertr r r i. [ ;.:.r. Frrc .j-r. 1 a i luifrC.r i -

( IL I t f ju..J r. [t '.I. L, 1 T, a Cn r r d ..:i.r. '*1 r.ure He Hcr rurj I ll:

S.tr, a :;i .: ,-,j, ., Li'J, r..ciL ,e,'. ir;T. t .: l : i[ l .:,,Jr .*i i d I**-.[ 'rd : i(

,r. p.1 ap Cr ..r h r..n e .. -cr.. char act r I ti C; I E r. c.5.Cc. r-. ..E ...d

r. .[ l I r uC : I E c a l :.r p. r... C t l t.-. 5 l ......l ..J i L ,-

I c .(c l >1 urn I n e r ee p. e- rr ad .* .-,r. p.-r c '.d F Lc

I I' : :. d Erh, it raCt .' .i ;zarl~.. p .r Ilj l ;( r... .. 'I Lul : .3 ,j-

. Cn;c .[I ,d rc .. d jlu.Il y1 [ ''r[. T i l a.iu I 1 Ee l.. r : I I ... r. 4 r..

b.;d, ija,r, r r .; 3 ,. i h 4lj ''' 1 e.-. j e r.a .

T I-. fi-ccts of irradiation on semen characteristics of animals

,-.dr ;.j[ r. t ..rn l stress of any kind ..ere not available.


Errr: .-r irradiation Compounded with Vitamin A Deficiency

Tr.e :-,, ,ork found in which irradiation was involved with vitamin

; *.t e,,ic...: clone by Meacham et al. (1962) working with swine. Sixty

cr...tr .d .... c.re allowed to two groups of 30 each. One received a

L .i- r ;..., c:.-r ii ning only traces of vitamin A, the other group received

rr,. :.;.c r t.I:.-. pplemented with 2,000 I.U. of vitamin A acetate per

.ur.-d 1cr leJ. .rter 13 weeks on the ration, 10 animals of each lot were

..:.. i r,-,..... ior irradiation. These pigs were subjected to 350 roent-

*j;.-n i.: ti.-.e-.-.d gamma irradiation from a 6,000 curie cobalt 60 facility.

i, ti--. lid : i IS ays, all of the control and nine of the 10 vitamin A

deprived animals had died. The average survival periods for control pigs

and those fed the low vitamin A diet were 234 and 286 hours respectively.

Other than this no variation in irradiation response could be attributed

E.., vitamin A intake of the pigs. The major :,"..;: Ts of irradiation damage








18


were: depression, skin hemorrhaging from eyes and mouth, increased res-

piration, mild muscular spasms just prior to death, extensive hemorrhage

of lymph nodes plus varying amounts of hemorrhage in the kidneys, heart,

urinary bladder, intestine and stomach. Shirley et a. (1962) in conjunc-

tion with Meacham on the above experiment found that vitamin A supple-

mentation decreased the deposition of copper in the liver while copper

supplementation increased the level of vitamin A in the liver. Gamma

irradiation was shown to increase the deposition of copper in the liver,

but had no significant effect on the deposition of vitamin A in the

tissue.














EXPERIMENTAL PROCEDURE


Materials

Animals

The animals used in this study were 48, 20-months old Florida

native rams from Mr. L. A. Maxie's ranch in Frostproof, Florida. These

rams were brought as yearlings to the University in June 1962. At this

time they were placed In a preliminary experiment in which 12 received

5,000 I.U. of vitamin A per head per day. The remaining 36 received no

supplemental vitamin A. The rams were fed rations consisting of 25 per

cent cottonseed meal and 75 per cent white corn meal. All rams had

access to coastal bermuda hay at all times. This preliminary experiment

was continued until January, 1963. At this time two rams were brought

in as replacements, from the University Sheep Unit for two that died.

From January to February the rams were given a period of adjustment to

a new purified diet to be used in the study here reported. On February

2, 1963, the experiment described henceforth was initiated. The average

weight of the animals at the start was 87 lbs. with a range from 115 to

74 lbs. The twelve rams that had previously been receiving vitamin A

were put on the control diet consisting of a purified ration shown in

Table 2 and 3, containing 3,000 I.U. of vitamin A per Ib. of feed. The

remaining 36 rams were placed on the vitamin A free diet.

Assignment to treatments was accomplished in the following manner.

Vitamin A deprived animals were not assigned to a treatment group until










blood analysis showed 15 mcg. or less of vitamin A per 100 ml. of pi:-i,,

A liver biopsy was also performed when such levels were reached. W-,.

12 animals were a.:cu.-rji.l td -hich met the above criteria they were their .

assigned at random, two to each of 6 treatments. Treatments consistiJ :i

an irradiated group (A Irradiated), a second group received 60 R.U i

PMS/100 Ibs. body weight administered daily by subcutaneous injecti:.

(A + PMS). A third group received daily intramuscular injections ,r .ci-

tosterone propionate at a rate of 10 mg./100 lbs. body weight (A + 7-:-

tosterone). A fourth group was given daily capsules of thyroproteli-

I gram/100 lbs. body weight (A + Thyroprotein). A fifth group was l:Ica.c

on the vitamin A containing diet (returned) and the sixth group was cc..-

tinued on the deficient diet without further treatment (deficient).

At the same time rams that had been receiving vitamin A wer. ..i,

randomly divided, two to each group, a control (control) and a grouu cp.:.:c

to irradiation (control irradiated). In this manner eight treatment jr.:..pi

were formed. This procedure was repeated three times so as to alloi rjhe

experimental animals in their totality. The experimental procedure ;i i:r-

lined in Table I. One of the animals assigned to the deficient group j-e.j

in the biopsy operation and was e-,.ludEd from the experimental data.

The experiment 1w. conducted it the Physiology Barn at the .ii.r.er-

sity of Florida. T'. ara .:.i the bj.,, used in this experiment cont in l

8 pens, 12 feet wii anij 4?8 rat i.iy and all pens had concrete fl :r:.

A metal rof covered tre fe-. arca j,.J 12 feet of thp pen. The peni .-re

ij j celr I pl, d n.l -:i.. ..-.r t ar. served two pens.











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'0 2




















































04










Sc aIe. and ...i:.. k r,. g rea *-' r, .. I. 1 6, Ie i r. h: 1. i 3 r. dl'E (re

, p:.r E t, I *ct- c c r. t tC'a u ; J.1 I rr- E h.E






The composition of the experimental rations is recorded in TaIte

2. Solka-floc was used to supply fiber in the rations. Corn oil was

included in the ration to supply fat, to improve palatability and to .r.-

sure a uniform blending of the fine texture ingredients with the coar:er

materials in the rations. Casein was the protein source and corn sugar

and starch were included for energy.

The mineral, mineral premix and .it-;in mixtures used in the ra-

tions are shown in Table 3. The mixtures containing 13 essential min:ral

elements was essentially the same as reported by Meacham (1962) and T.It.:r.

(1962).

Vitamins A, D, E and choline, in a corn sugar carrier were ad-

ded to each ration except that the vitamin A was deleted from that of Ir.

vitamin A deprived sheep.




Management and Feeding

The rams were fed dal., t t''u a.m. all the feed they would cliar.

up by the next feeding. Feed refusal was weighed back daily. Fresh .at.r

was available at all times. The rams were ,nrit r..d in the pens at all

I; r.i: -e epc r.X r v....F c to collect experimental data and for daily ,ir-

T-r. tr r.eC,.t T.er:- was one pen for each treatment group and six r.ja,,


























Ingredier


Cellulose


Casein (9


Corn Star


Corn Sug;


TABLE 2

COMPOSITION OF EXPERIMENTAL RATION


It Per cent in Ration


S(Solka-floc) 20


90 per cent protein) 20


ch 23

ir 25


Corn Oil


Trace Mineral Premix


Vitamin Pre-mix


Minerals 6.5


100.00


aControl group received 3,000 I.U. of vitamin A per
lb. of feed.










TL-ELt .

Jr .I F 7iE iLn 'i l i.J' f L 1MI n '1., VI T fI rIr
tj -Ti.ihL T i. :El:. i E* Pf A i NTM L P .T li :


L,', I H 'J
r



ril. i
FiaC I

B r, o.:r al f r .,-


FN : '. H H 2. '
N. 6, .:' Ir. l ... .

S ,,-, M .

I 'H H











i r* ..2..r lC'rr c'


l U .[ ..r. r ..




S ti . I n P 1 :.1:.':1 I


.. I 1 i r H r ,le |I'


I- I ,: 1 I '7 it, F. F ,


I;-' 6

*L l.-j It.: l. -r












I. i,
.1 :



11 r.

H ,02,










J ..:,T 1

i l Ij I

;. := tr :e i.[ LY. I r~i r


LcT.rn *...r liar r


?i..

11 . .







1" .








:" ':'0':' 'J' >'iu I, l



''. '..'

.:' ': : 'b .


Il.,c-- r p u-d u.* J [.. -, c .r, Ie i u. td : rcc 4

'L- r.. - i-, -i [ :.. *c .. ..c .r, ,3. e..J

*%jJJ ?i y., *r u ar [: [ I,, . .C. ..r... -. Jhir, ed r-.l .. r


_ I __ ~











y. I .:. ... r cr. pc... T .e r .. a .* .. .ed -: r I -e i : dJur ..

ti-c c*p'r: r -:,al v-ru d

TI,- th.r r c.c: I' .e 1 Jililu -r.'d 1 4 r.v ,11 r :.[I r r r.I T.,1 a rI:I ,-


























Semen was collected using an electroejaculator. A monthly col-
u j I r I E Ir.c hr1 v I .: -d1 1 E r ,I r
























election was made on all animals prior to allotment to the eight treatments.

A week prior to assignment to a treatment semen was collected eight times,

four times in each of two consecutive days. During the first week fol-

lowing initiation of the various treatments, s.en was collected four times

in one day. For the remainder of the experiment semen was collected once


a week.

Volume of semen was determined directly by collecting the semen

in graduated centrifuge tubes. Motility was evaluated to the nearest


5 per cent by microscopic examination (X430).of the cells with progressive

forward movement. A blood hemocytometer was used to determine the number

of sperm per cubic millimeter. From assignment of the various groups to











their respective treatments until slaughter a per.'.j ;r .r.,-.:r, ,-

elapsed, during this time semen collected was evel,..j- d F:r .j.jra .j il

data on percentage of abnormal cells present at ej,:- -. -i1 .:: ..


Blood Evaluation

Blood samples were taken initially and at .-rcrr 1. .:.i 8 o ,.

until the biopsy was performed. Blood was evaluar:.. r,,,- .,r ,, Sc .:,r-

ding to the method of Kimble (1939) based on the ,: r-. ... r- ,. ,,:


Liver Biopsy

In order to have a more reliable knowledge I I-- cr- r..-r..

of vitamin A in the liver of the animals to be as *.-.:.l r.: in. ar

treatments a liver biopsy was performed prior to r. : u, .-,, .,,

was done by Dr. Fred Neal from the Veterinary Sci,-n- ['p -f-r 1 :.rt [I-,r

University of Florida.

Essentially the technique consisted of thr. i .: I j

teral incision was made in the area immediately b:-.*-j [r5 i :- r r.-

right side of the body. A piece of liver was rer.:.-.: L, i.i :u ,. -

size as to allow vitamin A determinations on it. '.: .. : .. i r,

closed and 10 cc. of penicillin were injected to i--.*. r .,-r ,-, -: ,-,


Slaughter Data

On the twenty-first week from assignment E. i aC.t C u* :.. -..

rams that had survived were slaughtered. Of the i- r,. ii :ll :E C d I.. he

first group during the last week of April, 10 sur.. -~ .1r.3 m ir.; :mIu lrld

in the second week of September.













.it [I .eC,.,oJ Or.,,: r F r ,; ,u....- It., i':. h*j d t't.-,'. Il ,: IrEd


i tr. 1 1.1 11.r ..r .. rr. i n [ r-.r i I fcr .. -F C : .r.0 *..c i t : ---


tr Tr, r j Cr,..u r 1.j Lieo E1 3 : II 1- [ i :.C-:r. E 5 ..I JuI a .jk .Ej:


:Ijui"i c l r. [tI. t r. : D. E rec t r r rl ,, dl . I:-i. c .u '. ' J


u.L Il Fl r .T i C. ut [ .. rjcr .c. r

i- c,[t. mtilk i. t ,.j f. I, i C [ r r .- ..-. .. I .l c


per* 1:I [ rI c .r r ig.J dy r. .i- e re Ee'c ake.


.1. Ij ,.- (cr .i. i ir. rl u . LI 'J "e" J r-r r',r .-






Ti, i I.. S. e l. I 1 J i r .r I I *.: I LI r. jrI:.e : .:

























,., ,rCI .,2 u I I I.l i,, I ,l, :i *r I, :E : i.r i, .u Ii Ic- ,c j.[. i .
[ 'r., : i -d C[ j .[ l r ic r -e e F 1 F I r t. I:.. .:5i : i. : t.1. [ C..
r i.. i [ hdl .c r IF .i-F :'-[ ir. c t tcr dI i.



H EC c I C al rc n r I : -i-4 i i


T .: rI.[; I r r* [ r :E[ : L t .* =1 p C :v ttu E I E re F E E .













Tl.: :.r : C^ re ju[l l CJ L '.l. .. Er.1 t r. IIl : '.1 .0 t l J [C Z uJ,





tll r . -p* : C alu i .- r .v. f A (Ii rI f 3 -, al r al I E i ,-E- h-h1


*-rt t i .: 2 r.1 [E IE e I :p.,T,4 ,r : r r .i :AC rc r :.r, r .. uc .E r b..-': E .[* E ..-j


[ f. r :.i T ..- J r 1 r [ 0E I I r .. [r.4 .: . r. pc pi". 1 -r.u J .r


E I *J,.*C jE C j r .r. l il :.r ..j . Co : [ : m.













' I. -i.L As 1, j.

The Jd .I ..- JEl : ,, r : u'.'. 1. .-1 .*r,[r.. E1-. I p .1.1 P .,, : iI


.-..1 lF.. -:c I f -..C II h- rc .TL.r..r J r.:. r. 3: I (c(. c.:fI 6iCI,


Pr' s I c *E l-.Ji t. F .. [t.. 1- E .: l aCi.u...6I .i [.,EI ...(. ,lrc *u 1r a I 1 .




.1 tr _,-. F,c c.- .1 Prj:, I ti r. tec, ..3' .-n .- r [I., .- -, e.'.r i rF( r _1.*








I, rhe a ,. I. i :.r. ...e ..r. i., : ..._r, jar a. ..i,Z .J I..r *-eri




1.11 I 5 -,.1 I. [ 1 I c r.-.. -I I1 I:.* l. in l cr. 1 [E .. r- .r : II ., .. k : ..I


It,.Cc I' -:*c zr,, ,- .J .i rI : I. r, -,lu : r, t u Lt.r .pr.-:.:r ,,, .. I- .*-.,u


CLe r.I1 It,' I .I l : 1 J .c, : .I -rI[ E r. at : F C C I ..51 ,., .c *. , C S C -

I, : ..a.. r 5 C I.: r .. u r u.1- ar.. t c '1....c .. i. .


t :r.,,.: I 1,rt ..,,'( r, ., j L L, II. nE-... r. r ': -










I r. r. .. [ I .' E l r.,' .-..), L t lcr, l-cc -. c r ..r <


LE [.,,:r, [rcur ...,r. .re r .1 .,,'.co L, [ .: .[ r . rcC TK pr.: JuC r i ..1














RESULTS AND DISCUSSION


General Symptoms of Vitamin A Deficiency

Approximately two and a half months after assignment to the puri-

fied diet night blindness was observed in some of the experimental animals

on the vitamin A deficient diet. The rams were checked for night blindness

by placing a panel across an alley after dark and checking the rams indi-

vidually for their ability to avoid it when driven through the alley. All

animals on the deficient diet eventually showed symptoms of night blind-

ness.

There was considerable variation in time of onset and severity of

symptoms. Some rams showed a characteristic cocking of the head to one

side similar to that described by Lindley et a. (1949). A number of rams

showed a characteristic cloudy cornea. In some,this cloudiness disap-

peared but in most it persisted. Figure I illustrates this cornea con-

dition in one of the vitamin A deficient rams. Most rams on the deficient

diet eventually exhibited an uncoordinated gait and progressive weakness

which became so severe in some cases that they were unable to stand as shown

in Figure 2. Several rams walked on their knees, apparently as a result

of loss of control over the lower part of the leg. This condition was ob-

served in only 3 of the 35 vitamin A deficient rams. Other symptoms exhi-

bited by the deficient rams included loss of appetite and evidence of pain.

It is interesting to note that some animals did not show any degree

of incoordination even when the liver stores of vitamin A had been nearly

depleted.










'4


Fig. 1--Cloudy condition of the cornea present
in a vitamin A deficient ram.









A mr w
a---
.. ... r... .


., .' i s .. - -

Fig. 2--A vitamin A deficient ram showing extreme
weakness.


A 9, _OAW' J00











The deficient rams that were returned to a vitamin A containing

diet recovered from all lesions. However even though the vitamin A liver

stores of these animals had been drastically reduced no severe symptoms

were evident in most of the rams in this group at the time that these

began receiving vitamin A. One of the rams receiving PMS treatment exhi-

bited a protruding eye with evident infection. At autopsy the veterinarian

diagnosed purulent pleuritis as the immediate cause of death. Another

ram on the same treatment developed an abcess in the sheath area. This

same ram showed bloody urine before dying.

In general the rams on thyroprotein treatment showed less severe

symptoms than did other deficient animals. The appetite and growth of these

rams was higher than in other deficient rams. Rams in the A + Testos-

terone group exhibited symptoms similar in nature to those of other rams

on the vitamin A deficient diet.

At autopsy no striking gross pathological lesions were evident in

rams subjected to the vitamin A deprivation.

This author believes that some of the deficiency symptoms observed

were in part caused by multiple nutritional deficiencies resulting from

the decrease in feed consumption observed in the vitamin A deficient rams.

The A- + Thyroprotein group had more appetite as well as less severity in

their symptoms. It should be stated however that the deficient ration

would have supplied the required amounts of nutritients had they been

consumed at the same level as the control ration. The basic cause of this

multiple deficiency is therefore an absence of vitamin A.









Entv i f'CL i or C jr. irrsJ;Er;.r,

Rams exposed to irradiation suffered a gradual loss in appetite

until death. Those rams surviving the irradiation treatment eventually

recovered from the loss of appetite.

No signs of external hemorrhaging were evident in any of the rams

exposed to irradiation. Several rams exhibited limited anal bleeding

prior to death. Most of the rams would lie down, shiver and show signs

of pain prior to death. At autopsy multiple hemorrhages were present in

the lungs and chest cavity areas. Limited hemorrhagic spots were observed

in the cortical region of the kidneys and adrenals and in the urinary ...lj-

der of some rams. No characteristic histological lesion was observed ,r.

the testes of animals exposed to irradiation.

The multiple external and internal hemorrhaging described by

Meacham et al. (1962) in pigs exposed to 350 roentgens of whole body

gamma irradiation from a Co60 source were not observed in these rams

suggesting different mode of action of Irradiation in these two species.



Plasma and Liver Vitamin A Content

Table 4 shows plasma vitamin A values for rams in the various all,.-t-

ment groups (Group i, II and III) and treatments, a summary of all treat-

ment groups is presented in Table 5. It should be noted that the various

treatments appearing in Table 4 were not started until after the biopsy.

However the rams appear grouped into the various treatments so that compai-

isons can be made on the vitamin A reserves of rams assigned to a given

treatment. Liver storage of vitamin A determined on biopsy samples is

also shown in Table 5.























- I C C N -





















4
- r I .



2I 4 C -. 2 7


Cr




















- II -l: ~ 7' - .







-3 -
















-Z -.r~


-T C S


Zr' -Zr







C-
\I J





9




'' ''
















4 i


i







r


b-





_1


.i .n











r






c




















i


-


























1


z


? i


'-

















TABLE 5

AVERAGE VITAMIN A PLASMA AND LIVER VALUES FOR THE
VARIOUS TREATMENTS AT TIME OF REALLOTMENT


Control'

Deficier

Return

A + PMS

A + Thy

A + Tes

A + Irr

Control

All vit;

All vita


Number Plasma
of mcg./lOO ml.
Treatment Rams of plasma

6 36

nta 5 3

6 9

a 6 5
a
6 3

t.a 6 3
a
ad. 6 8

Irrad.a 6 57
b
amin A normal 12 39

amin A deficient 35 5


Liver
mcg./g. of
fresh 1iver

154.0

1.4

17.8

2.3

1.9

2.4

7.7

157.8

156.0

5.6


aValues represent an average of all three groups for
a given treatment.

bValues represent an average of all three groups for
all animals receiving vitamin A in their diets.

CValues represent an average of all three groups for
all animals deprived of vitamin A in their diets.













T ; I I.-. i re . vr. jII r1.- .. :; E h C J I't-r- E r. II 4i .-.. I

Ti irv c: r peered IIu n ..: r [t-. r. -r r .:.&- E -, E[ IT.. J i ..Cr -I

' I I r -. i...-. [ .1-. II: I ,r l t . : C E I I C: J K T - e j.,iu

I 1f h*'.. v r E t i : >1 -: ." ,- 'I.:...- -. DC.k p I *-.: it *I I I E f r. r. ..

T i.e J I r I fi rr r.r .r. r r [ a ..I. i ij r E .r. [ e r .,..: a J.:., t. C j .:. rr.;

.:tr ..-. t I t : .ir t aL i .

T ti I I:. rt-..[ E .: r i ..I I, '~ .. .r ,...,,-, I .tturrin j

from month to month ir. 3 ji --. ia r f tas . : c*.:. icrat rcr.: ii -V.Ecr,

the plasma vitamin A c.e I : .a. tee... r ri c.C j I i .i r. i ri. ., i i. r

stage a plateau is reachej. It E as at L L :,-, [ .E rr.h c t :.p ., p Cr-

formed to ascertain the deficiency conditi-:r.. I .i er.ral ..rI ...ir. li.:-r

reserves were at abnormally low levels at *--: 1 t ,v jnr.r :Fr n- re".

on the deficient diet to the various treat-.Ei : .- ;r,.. I Tre 3.7:

S : r. c.j [... r.. ri. tire rc r..r . cr-:.p III : e.1 K E. 1.r .i S. .. -

*r Ie .jr, :,[- I r ..r ii.3 ri T but C**. J. o t, I I .i [E h r. i- :.:E re : .-.

-r, f.. *...-.L i ; rn:riana l rr l. .T. Ir. j .er l ..*r.e Iea 51 r. .rate I l

a I S r.-Is iC i.r r-.C C.:..d r, .5 r -r.[ :. r t.. I. 3- C1.. ..T.-.t I.

[tEh r .; C:... [i. JC T *ci r.r .J E[ (.:. [Cl., ..... c rca ie.s[ r p- a r II. I I

: "- Ij kc= .; 1 nr .E, tE6 -nE t. t.. (i L E .E :. E E - r,v .cr I i r

A 1 .I : r. 1l. Tabiie \- Fr- r a h.rEj 1r. Eh.ese raT uk.1i .-i .I -C tC E

* riL I j re, j T 11..3 :. p or .:I :.r:Isr *r 1 :ir. ii[. ti [i -e.. ;i n.ia (.-.

r. Ir ir .i A ..u e The- . l pI : i .I a, r=: b:C r .. 11 r..-. ir., i-...

I r I .' i.aS Et -. i .ufld i[ r.i .:.t rm ll.SE-rnu .,ert- rEc he .'C I tu d

r.acje teen. r-:. pEri :...- i r :I ,-s :c T .: c E i jr. tLi cad.c i ; i. I al r* .cr l

l-rc-ugh-":'ui c- pir r. E TID .: .: .-: r,a: bie c ( i r *: .j..? (rJ












. : [ I i r.ke e .,. I (. 'IL-d ; l h.L a . JA=5 r. I : .e ,. It I 31 :*:. i: : : b0 t

IJughttr rf 3.J.. ip- .i l dj t c trerit per;ud: and ta ad :.; r, [r.e amu l

nrc r i aTa, r t.t atu [. ift .pula., t, 5 c -, 's,3 .1,e t iG t Hi; : .. r E r. [E l r. :

e p-.rir...i ,, ( t h I t 3 [ er .T ths t uI1 &r, I .: I low.. : rc or .I/ j i t.Tn ot-.

nriter .- r 1 m- were ca. Ia lat.lI

Tartl. 1 ,: r Lt tap, ,, ci n .., i ,r i ,, J duC u r I IT.T- 1 31 -.3 ai. i l i . r

..[a.T.i r, r .= E ra?. [ ..C f r.-a I .i trr.rr.. a , Tr tl, e i ,n- or. ir -

%1 a l t I .I 3 . .[3. 'I. .i .- ;3 I r .-r .. ,,-| _r. r ,: : ,_ r, I. : r r,-

a. 3 . thn C4 the pe .nr ,.c The p l3T- .13..,. 1 .E1u: ; it..ui*d pr c E:r

*r r, .,-,: .. (.he .J, ,. -, It C n c 1 t r t Ir t .:. i e, ,, .:r. Jr I t-a..

E .: :c i r Eh, 5,up ar.i In I- ir re ur .e r up r.a ra r. ( 1 1 .7.

.Ia -i Iu- 3 r lab I e tab thn C u.l r.tr.I:.



Fed. IE 'c r

Fted a .( e ca j .; r i.-1.i dual r a I r er. :t p.: L tcE t- 3Fc -r

L I-u.c c i i i uI E .- E id C l c. er 0 ii0 .I ; I.j r.-r i.t J uCid.l

3111 [r..C, E Dur lr ] i tgra- II Itrd .n j m tt Jji j rt I r rt:-r .:.rir..:..Erdi E. tc -

ca [ n a *:.4n- nt r vn. :-. n, .:. the i t T.--n e : .13. ; d..r. ii-.. .ret

lr :c. r, r.3 C cKr. ejr' held 1 -.rcna. l, I .r. r I r.uT.CItr C i a-. ur iI

thr. 4,7 >er a .-n eC 3 c' c .-- Itu. : hlt wae In.- -.t.er --.f ra- 7 ma: .-

Sre cn[ ,. l [ ..I E p r..':-:.. t .:.r .r e .: It h c t re a .. F: r

thic- r- a: i : cal l an ,-al -1 i E I rI t .k ,e .I a t t c M*A C : t-I

h er .. r -.E -ir i-I c -Ac ;f. J* r. Jt CL .: rhic : j Cu.c c I u : : i. an t dr n..-

Fr Th [t.c 343i I it Jat a

1 3tbi. :U h' W c fc- .J I :.t djaE a r...r (t.t .ar ,;.u tre -at.rn. t jr,-up

*ur ilj prd-6: I .1 11 I pha :i I r a,-: .*r [.t-c E ..C ,I 31, j A[c i;,1-,











J
P


- 4 -


0 CO 0 0
r --


- - 4 -


0

0


-~ '7 0 7 ') 7 3


44 0 0 0 4






-. 3) O4*~ 4


'3


4-4 4. '


44 '


*2 3




'3 '


- o


3T


-, -


3) '3
C LI







o C

1'1 4-1
.._3


0)
o
.D



ow o
o .3


S



( a )
C 0 -

C O 4
m 0
c- m
4i .4- 0


3) '




4< C 3,
S o S




4 3







*0 0 4-
C a3 C
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o< 3, 3,
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70 0 .
^


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34. 4) 4- 4 O

'3 0 3)










deficient diet consumed an average of 3.0 and 1.8 Ibs. per head daily,

respectively. During phase I no widespread occurrence of vitamin A defi-

ciency symptoms was observed in the rams on the vitamin A deficient diet

indicating that the detrimental effect of vitamin A deprivation on feed

intake was manifested prior to other external signs of a vitamin A defi-

cient condition.

During phase It average daily feed intake per head was 2.1, .85,

1.90, .92, .71, 1.59, .67 and 1.54 Ibs. for the control, deficient, re-

turned, A" + PMS, A + Testosterone, A" + Thyroprotein, A + Irradiated

and Control Irradiated groups respectively. Rams receiving the vitamin

A containing diet during phase 11 (control, returned and control irra-

diated groups) consumed more feed than rams on the other treatments, ex-

cept for the A" + Thyroprotein group. It is interesting to note that the

control group consumed less feed during phase II. This resulted from the

increased stress to which all rams were subjected during this phase. Thus

the decrease in feed consumption evident in rams on the vitamin A deficient

diet can be attributed partially to the increased stress. The A" + Thyro-

protein group consumed more feed than other rams on the vitamin A deficient

diet. PMS or testosterone treatment did not have any beneficial effect

on the appetite of vitamin A deficient rams. Irradiation appeared to have

had a depressing effect on feed intake on all rams irradiated.

Feed consumption in the control group was comparable to that re-

ported by Meacham (1962) but somewhat less than that of Tilton (1962).

The latter used corn cobs as a source of fiber in a diet very similar to

that used In this experiment. Meacham (1962) used Solka-Floc as the source

of fiber which is similar to that used in this experiment. At slaughter,











extensive amount of wool was found in the rumen of rams in all trcT.--rr.-r

This was to be expected from the frequent "wool eating" obser..i.r ;rn r.t

experimental rams. This suggests that the amount or the source :r r,.cr

provided or both were not adequate. The possibility also exi:t: tr..t

some other nutritional factor was not adequately provided.


Body Weight Changes

Table 6 shows average daily gains during phases 1, II a 1.d I .J.1

II combined. In phase I rams had not been assigned to the vas..,ire tr -

ments; however for the purposes of this discussion rams that ,irc .js-ij-i.e

to a treatment during phase 1I appear in this treatment in ph.:= I. :

that it is possible to follow growth rate for rams in a grou: riir E.: jnd

during treatment.

During phase I average daily gains were .24, .06, .10 ... .

.06 and .23 Ibs. for the control, deficient, returned, A- + PrI.. A- Ti:-

tosterone, A" + Thyroprotein, A- + Irradiated and Control Irradl;aed

groups respectively. Table 7 shows the ranked average daily .:.-.: Fr re.

various treatment groups. Rams on the vitamin A supplemented diet jaiwd

significantly more than those on the vitamin A deficient diet P: .,a- E...

be expected the various groups on the vitamin A deficient dire dJd .-:.

differ significantly between them. Individual daily gains arnd .,irgt

during phase I are shown In Tables 19 and 20.

Average daily gains for the various treatment groups d.urnr. prasi

II as shown in Table 6 were 0, -.10, 11, -.04, -.27, -.04, -.2:, 23 Ib:

for the control, deficient, returned, A" + PMS, A + Testoster.:rn *

Thyroprotein, A" Irradiated and Control Irradiated groups, respecii..l,








































~I















U U.





I-








CU


O
L
Y
E
O
u






o
P


+
s










+ u






o
< 0

c






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Tl.1i 7 :":h o (rc r if 3.Led j.ir g ja jI .3 -. r.:. t hie :a.c grr..u : I.e

r. -tur. ,d 3' .ul c .; dl rl d ll C n I, r C [ P.1 ai I .[r.[ r r r. =i r.. L r..JP:

? *C pl t r.,r he c-, ,.rFrol Tr-. crnt i cI : d j -.it -g a ; :.gn i r c ar r,,,r

[h 3.1 Et.r. iM T ,rc.pr.. r.; r, ari.j et ic .-.( r Tu r.T, ? I 3 ,-r c

da I I a'. 6 re C ..: tr*.'l r ,m,-, : I v r)1 l lut, l [t -e Jrla : ight reJuc -

1 1.0r. r T t* 3 .h.:, i. .i i I b dur r.g ph e II ia. C.,: :cd .:

r r ail i i t! J -c r .a I. C cri a ci Jal i .arn fr, lw fcrt I. 1i3 ci ICt A I

irradiation on average daily gains was equivalent on bo:r. c.:r. rcl Ir.,d "

Irradiated jr ,>p:. Individual "..ei l. ; and average daily' gairi ir. : .:-r.

in Tables 19 and 20 of the Appendix. The combined phasv I anj II a.c.r n

daily gains are shown in Table 6. Table 7 shows the ra-.ed a..rge j~1 l ,

gain for the various treatment groups during phase I ani II c :it..-,.d. Ml

rA.7: .: .- lhe ; l 3>7. r, :upp't..'etret d I5 l .-. i l*.-illcautl rC.C re 1,I..I,

r ,i: s ,-, hF e rdi ,FC i il ,i rat li I ii r .- Ipni l* thl : i e l .idu l




t -,.,I : : r s r i l. [r te [rC b.:. -, 9rg t cr.mr- .a3 ir r. T.i. ,.r,

[r,. ar-tiu ; tueat.T.e i v t..ri d i :,u: I.4 thci : r i uri e .[ :h,...lId be *.':leJ

tLI [ trE e r.....L r oir r 6rr. pct (rea[r..cr-,i ., :,, ll frLrJ.-. the a. (it, p.sr gr.:.up

a i. Cr.s rf., d ,r, I.1:- .r,rti tce.j I, r. T...I A L, C d rici.cn Jr : tica l,

r.*jd-,C J tie r.'11: p rr 4r-K p :-- Tha* -I r, .r.j ir- air.ce5 il.,- 5 ,e p [lcted

ir .3 rre I E .-. J( rr re t:r.( [t.E t.od, ofe.oI ch ari gce 0t ad .er, I *.i te d

nfat.Er .EI rA .-..: I Lr r -re.., cr C d3c r. i[aI [ d.d L t C.i.e Lr.-.:r.A r,

,:t c. e-.,aer r..r t :I In., .r.. .ad ,.., rty,. ne ll-.e rcu t r :.,[ -.'. i 1. t

Cu r. t t I I tie-pe .-j Ci.i p clrl .j I rme *.,.,ld har- Eari :I[eep r I.ad [ C,

'ur i .Lj.


















PHASE I





6




5



./^


PHASE II









/
..^/-, "


3s'


2-








DEFICIENT ------------
RETURNED ------



-40
3 6 9 1 4 7 10

2-WEEK PERIODS C

Fig. 3--Effect of vitamin A supplementation on body
weight changes of rams.

aphase I extended from the start of experiment to assignment to the
various treatments. From periods I to 6 there were 6 rams per group. From
periods 7 to 8 there were 4 and from periods 9 to 11, 2 rams were present per
group.
bphase 11 extended from assignment to the various treatments to termi-
nation of the experiment.

cNumbers adjacent to plotted values represent animals remaining in each
group for a given two week period. Differences in numbers between periods in
a given group resulted from death loss.


















40-








20-






o
L.)
I
0
I- -


Lu




-20-








-40-


PHASE 1I






6




S ,, 6




----
6












CONTROL
A-* PMS
A-* THYROPROTEIN --
A- TESTOSTERONE- ----



3 6 9


C
2 .,EL.- ER ODS


Fig. 4.--Effect of various hormone treatments on body
weight changes of vitamin A deficient rams.
aPhase I extended from the start of experiment to assignment to the
various treatments. From :er';.d: 1 to 6 there were 6 rams per group. From
periods 7 to 8 there were L ,ni Irom periods 9 to 11, 2 rams were present per gr;uo.
bPhase 11 extended from assignment to the various treatments to termlna-
tion of the experiment.

cf..-,Lir: aJ;acent to plotted values represent animals remaining in each
group for 0lr. nc..-week period. Differences in numbers between periods in a
given group r.:ultEJ from death loss.


PHASE IIb












5


\ ,










4

\ \



2\.
\ I

4 7 10



















PHASE I PHASE II b
40-








20- ----





5 .
m 7 ,
u 2



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FgI 5.--Eect of irradiation on y ight h es o Gtmn
-20-
CONTROL
CONTROL IRRADIATED ------
DEFICIENT IRRADIATED -- --



40 1--- i -------,- ------------
3 6 1I 4 7 0
C
2 WEEK PERIODS


Fig. 5.--Etrrect of irradiation on body welgst changes of vitamin
A deficient and normal rams.

aphase I extended from the start of experiment to assignment to the
various treatments. From periods I to 6 there were 6 rams per group. From
periods 7 to 8 there were 4 and from periods 9 to 11, 2 rams were present per group.
bphase II extended from assignment to the various treatments to termi-
nation of the experiment.

CNumbers adjacent to plotted values represent animals remaining In each
group for a given two week period. Differences in numbers between periods in a
given group resulted from death loss.












F. ut. .1 1 u tr .,5 i the -e .-. ,.i ht Ch_.nc) i: th-I C.,. r ) Jcti -

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i l rI .: ,, ia ,' ior jp '. tlh 1 t4 . cE fpc i i ..J l. Lt..-: jinr [E r

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turr.e. r r.: I c I al :frr..-i T re., e I., en r a.-,: C l cd [. I .. c i 'fr.[

I.h I *:rc-i tr.,Ilr i ncrEo-i .d in -.i -r [ up t. fc J ,.j thr,-- iA, Ci-n: 3

plt -tc ,u F ur, :1- i, l-. L .. l, .- .hlit iCrt n- t jr [r.lc COr. l r...I ja r..-.r-..c.

tr- t A _r ,up:. ur i r,J i3r, : I Ktne :,, 4r, I r c .; t C rn.r :, in b,:. .i .r.c .,-C

TI l- C :-C-. r il r ,7, ,:C: .Tp ir .1 rt-i T, :.r. th-y d f.Ci-r.t d. t I : le rl f I t... r.

L .r ;.- p- :,. II rA.T.. .n t r. 4 PM'. jnj n A T .. ri n n r .up: urTcr;J

dr t:inr *t i a c u L r t I CC IEl.rti t i r r [ l r ic ir,.: tedn i;C dl ir, t

tt .:,r t ei n r.,d n,1 pr-i nn i L d igh l I : : ip [.:. P, r 1 r,,; ,.

part Ial ly -jj., t5 a iiT1 it :n in apijIr i prduCvd bL, i.t i(.,rricprn- C n

F i jrc Ii l r i ri ir c ct.l ..t ir r 3. Att ift I -.,- 4 L j J, .*e I P.t

C. air-, :. It CI ,,r I r.at al rI I a E p .d [: rr aj i I ., : v Iul"rri r e3 uert

-r i t In- -eL. Tr: :..rrel Ate .,11 .inr tr.e Jecrcadi : n .e c .n:.J n ,)j ...

Lie .ed Ir, [.r arE,: r.iT, Tr,'.e rI : ;-, ithe ..- r. I rr ad;atle3 r up .v

:ur, i rir .3 i a t rec : .rE ,J

T1 I.' re i .1 e c t r. g r..ut.l' ra i Ar,-a I i c j -e th ;t ..-.in

C31 im: i.: r ., . ...b: r.-e.l 1 t I e p r .Tr- r.t F -,r rj. :. .. t a .., A r : er e:

V- h d b .jr ia; |ic I ricduCECd r..j '. a- -.p reicr.4: r, i rdI -nl* l I .

,.er" stle t... rec.:. er (.en ),'CU 1.1.1 -r ta.i l .e r It. ..1 re-.j rr-

ip -. in jled. PM: ar..- t -:lt ):.erC.n. t r.Ar -A.r-t Jid rt.r pr e riE r Fturirh r ., i j h

In-: ; In t iaL.rr. Jdl i.e, tir ma,.. Tt. )r-pr...i :-pp- -jrCed I: I-. Ae 5 itI-

0iuatI nj CM' L unr. rpp- t iE c re mitlr l arP rt l i r. I c :c re rc- .l t.t











losses, as compared to other rams on the same vitamin A deficient diet

without thyroprotein. This finding would appear to contradict the widely

accepted theory that an increase in appetite which would normally sti-

mulate growth, if the diet in question is deficient in a nutrient would

accelerate needs for the nutrient in question. This would in turn increase

severity of deficiency symptoms. Irradiation affected feed intake almost

immediately. This was accompanied by a severe weight loss until death.

The appetite of rams surviving irradiation returned to normal and did not

undergo further weight reduction.

Analysis of variance for differences in daily gains between the

various treatments in phases I, II and I and II combined are shown in the

Appendix Tables 26, 27, and 28 respectively.


Survival

Average survival time expressed in days as shown in Table 8 was

141, 82, 141, 85, 97, 125, 46 and 63 for rams in the control, deficient,

returned, A" + PMS, A" + Testosterone, A" + Thyroprotein, A' irradiated

and control irradiated groups respectively. Survival time for rams in

the various allotment groups (I. II and III) was also shown in Table 8.

Per cent death loss for the same groups as shown in Table 9 was 0, 80,

0, 84, 66, 50, 84, and 66. Table 10 shows the ranked average survival

time in days for rams in the various treatment groups.

Rams in the control and returned groups suffered no death loss.

They survived a significantly larger number of days than rams on the other

treatments except for those on thyroprotein. This latter group survived

significantly more days than all remaining groups except for rams in the




















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PE- L(ElT UL-frI Lit.. Ful-F -Mi IN THE
Vit lO '. Ti.E.IMEILt


II ..r . r. l : 9- 4C ..-
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A + Testosterone group. There was no difference in tolerance to gamma

irradiation between the control and deficient groups exposed to irra-

diation as measured by survival time. Differences in survival time be-

tween allotment groups (I, II and III) were not significant.

Most of the rams whose vitamin A reserves had been depleted even-

tually died. Treatment with either testosterone or PMS was not effective

in preventing death losses resulting from a vitamin A deficiency. Daily

capsules of thyroprotein were effective in reducing death loss of vitamin

A deficient rams. Rams whose vitamin A reserves had been drastically

reduced but were replenished by feeding 3,000 I.U. of vitamin A per lb. of

feed daily suffered no death loss. The vitamin A status of rams exposed

to irradiation did not affect significantly the survival time of these

animal s.

Individual survival time for all rams is shown in Appendix Table

21. Analysis of variance for differences in survival time between the

various treatment groups is shown in the Appendix Table 28.


Semen Characteristics

Semen data %ere statistically analyzed during the first ten weeks

of phase II. Beyond this point total number of rams per group was too

limited for statistical consideration of the data.

A series of figures are included which illustrate semen charac-

teristics of rams in the various treatments during all of phase II. It

should be noted that in some instances the number of rams per treatment,

as Indicated by the numbers in parentheses adjacent to the plotted values,

is small resulting fr:e. h-a ,' death losses in those groups. It is also











true however that death losses in treatment groups on the vitamin A defi-

cient diet occurred when these rams were in advanced stages of avitaminosis

A. At this time changes in semen characteristics affected by vitamin A

deficiency had taken place. So that in many instances plotted lines even

though based on data from a reduced number of rams, do reflect the trend

of the initial population. For example Figure 10 illustrates per cent

sperm motility for several treatments. In looking at the A- + Testosterone

group one sees that from weeks II to 12, 2 rams died. After week !2 a

drop in per cent motility was observed. Appendix Table 24 shows individual

semen characteristics for all weeks. One sees that the 2 rams that died

were not showing high sperm motility, so that the drop in per cent sperm

motility at the twelveth week was not due to death of rams with high sperm

motility values but rather to a drop in values in surviving rams. Had

the rams survived the change from weeks 12 to 13 the slope of the line would

have been more pronounced. This should be kept in mind in drawing conclu-

sions from the experimental data available.

A summary of semen data collected monthly during phase I is shown

in Table 11. Per cent abnormal sperm cells was not determined during

phase I nor was semendata from phase I analyzed statistically.


Vol ume

Average semen volume for rams in all treatment groups during phases

I and II as shown in Table 11 were 2.2, 1.4; .8, .9; .6, 1.1; 1.0, 1.0; 8,

.5; 1.1, .7; 1.9, .9; and 1.0, .8 ml. for the control,deficient returned,

A + Thyroprotein, A' + PMS, A- + Testosterone, control irradiated and
























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.: r E..:.E ,:,n it.- i t ; r d l C.cI ICe r. d;. l dur ir., pla. I 4r T pr.3 I

t;. p I i f re .,h r : 3 dr.,,, 1- .er3.j :ci, -i .:l uT i. if. [r. : C :,Il r r ..

Th : 1: Tr ,ta I j l Ju I t.l u r i::,' .1 i r EQ u -. .,C .:, ,I ,cC[i .r, duI : :,n,,

. E I I Ti -,. iI .-C t j l dI J D E ** dr.... .. c a. r il .*Clui

*lur ir., plr I 1 II rir r. :t ur ,' 1 jr, Jp.C j, l : .. d :T,-r. ,lu '.; J ur i p. :. a-

I .. ufCr a.;ige c*...ld be a1Cr *itu d C-. j..T.cri..tf r. i. .. .. 1 1pp -

C E, at. :r.. i r 6 .II :t r et :e ner. I n lu7c r..r.ar : w. r r r.c r3 jrrCu

du.' I r.j t I I i ll i e r.,:. he :*-. rul .. d,: i a .T ..; r. d .d r.

crl- .iu:. b E-... [h .. tr :r .r uP ir,..: gr rua ... \ *j. I iet.- ri

t urrneJ r.ui C rlE har : h r, ICi (i. tC.:r, r,,,-r., : upr..li.E.rl, ed .j. r upi LF. V

PM' .,id 3 c: I ir .:.. t E I d r e Tc.; :ul 6 Cr d a. I ur j .r I e u it. n i' a e r a.r

.Cie -. C.lu dur. r. n t. .E II. Th-, *rcpr,: (e Ir, ren:.i.;. di d -..;.-: r. [ Ir

riarqr. .us c E Or- ,:-I Eilu.Tn;. i ur: Iu:rr i : -.-cE r. .I.lu.T.c Cr--3r46

r,.r the hormone treated groups compared to the controls. Semen volume

in the testosterone group was quite variable from weeks 12 to 19 but it

remained lower than in the controls. The PMS treated group had low semen

volume throughout phase II. The semen volume values for the thyroprotein

groups did not drop below their initial value in any of the 20 weeks.

Both groups of irradiated rams showed reduced average semen volume during

phase II; this change was more drastic in the control irradiated group.

Figure 8 shows this change during phase II.

It should be kept in mind that semen volume differences between

all treatments were not of statistical significance when analyzed. Tables

30 through 39 show analysis of variance for differences in semen volume





















































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during the first 10 weeks of phase II. However some trends were evident

enough to permit certain conclusions. Vitamin A deficiency appeared to

have had a depressing effect on average semen volume. This effect could

not be overcome with thyroprotein treatment. Treatment with PMS or tes-

tosterone resulted in a more pronounced drop in semen volume of the vitamin

A deficient rams, Irradiation appeared to have had a harmful effect on

average semen volume from normal and vitamin A deficient rams. The reduc-

tion in semen volume resulting from a vitamin A deficiency found in this

experiment is in agreement with the findings of Lindley et al. (1949).

They also found no beneficial or harmful influence on semen volume of vi-

tamin A deficient rams from treatment with PMS or testosterone. Goswami

(1962) reported a reduction in semen volume of bulls treated with thyro-

protein during the summer months in India. This effect on semen volume was

not evident in thyroprotein treated vitamin A deficient rams in the study

reported. Black et al. (1950) reported that thyroprotein had no effect on

semen volume in rams which is in agreement with our findings. However it

is very difficult to compare our data with those reported elsewhere because

of dissimilarities in climate, diets, hormones dosage and age of rams

present.

The harmful effects of Irradiation on semen volume apparent in

this study have not been reported elsewhere.

Individual semen volume values for all rams during phase II are

shown in the Appendix Table 24.



Average per cent motile sperm cells during phases I and II as

shown in Table II were 71. 72; 29, 22; 66, 68; 84, 48; 57, 19; 88, 18;











;8., l* and b61 46 r r the c...r, tr, d nc i; r.[ retur r.d, d Tr., r.:r.:-r [ein

~ +* Prn'. i~ Ie : tCr..-re, cc.r.trI Irradiated and irr alat-d 4riupi

re pecr .el ,. iLur..19 pl.a.- I LTr. c.:,ntriol.' rcup :t. .d Ii.I I I I E al u

ri hre r [, r r ,..,. Ir,-, [ct r i. 1: I r. 9 r uup. re:.t i t: lu : i m :t' her r up

, rh- t .il a n rA der.C I E n I e I c .r c C:rp.ar at 16u t t re c .3r. r:I .cf3

r..: III .alu.: t.:.r tI'e C.nt r l r ~;. ere iiilJar ;n pra:: I ar.1 I. The

Jet i ic n grup .u red urer rrtr : n r tir I I., ale jr. ng phjie I

.EraI c ..:-.ti i i .:,r the ret t rr..:-d grjup ii.j r.,C Le r, artEcred Juri.nq phaue

I :. It P il 1 i t l h.: t,e r.,er = dr..p 1 iotil I i r.. .I[E.& i r.

E t I ,C .E.c:/ C .. L :.be erc*.-1 C tv y raT.I r % 4 : upliceit. at3 IO.1.

Fi.Jur 9 i luiirarJ i cat ; ig n per cent rf..:.t le : pe... dJ.r iny Dh

II. ir. it.c c'.tr.[r.. d ffi i ;cr.it r, returned *gr.up: tscept ror C -F k f I

rt lit i alue r the j: l. :i gro up i re i:.c r ti-. a ( :-.: : f r e co, -

tr:-1 3.I c ri tur d 9r.:.up: Lr i l ierer.ce i i. per cent uoi i e Eperri tie ev r,.

tihe c..*n r:. i,-d dei ~fl er.t qr.-up : h -i.r. ir. T bt e I2 .. re : ; r,.rica.Ir in

, es: d. 9 and 0 or .1 n l;,.I: or ..arl arce t.:.r d;ileren.ce! per cent

m..;Ile :peir. during all 1 r I ie rir:i 1I'0 e.kz .:.t pha:e II are :r..-:.-. in the

Appenro.ii T I e 31) tr.rougr. 39. il>erage per crit iclit. [e iperac. aluu a

:i.0,,n i r TathI- II ,.erv loirc e Jurin3y pna:c II ini tre i'ori.,-,e trrealed rai..

TI hi r ..pru etei .-. Ireatr.Ent .:,jerc *Me r hir-.a t tihe dep re : ingy crr ctl: t ;-n ac-t i I lity

IrciT. i t.e .r I. le A lef I er.c, F igurc I1 il I trite. aer .ge p,:r cent

,tile :pErii r.cr tt., r..:,r.Tonl treated anri c)rtr l group: during pha'e 11

Frc.,i uEek: i ti: 4 a drop ,n j. Eraye per cint m.ic;le .pcr cell. a.: oterred

in all r r:..,r-ritn treated ygr,'uip rp iT ..eL.; 4 to II taer:e 47..:.till ue

.arit lr iJr. the A" TE: ic.t er jane 'ruup but ej,'itual l dr:-pp d lt. O r i T,

jceek: 12 Ct 21. Fro.i. -.;6i : to 6I jrer j.3 per cer.t -,.-t i L 'peri reiT.ained
























































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[urr ed (.ah .ch ha.1 r.i,l I I;s U uC C.Tii r atble Er : ,. l dIr urr l t ..- 'pI a t(1e

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qr.u m I 'ere C. [ i ati :Lica ll, iynq il caf nt ir. -ir v o r r... I i .- II) u k .:Q

phr.:e Ii 4 I ,-,a of a i rnc I i iferenflce r. per ce-i : ri7 r .: i i1t,

duri'Cy p-:at I I are -.o.r, I r. it-? App .-J Table: 31 Et.rO.a r. 19

SIul : *tL. A l ined in Ir.i neris.-eni -.nd:cate ithat :-cre., c: E; 1t,

e .c.rE.rd s ct C eC J bl .i iE a r. H f I C E.- ,. Tr,. I C- rej m per-. ..C. t -

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ufluC:CC ui iin pre.enc g r I.Tipr. s n, p r. |prr 1T.D ix E, a ....r, P

dcr icient r a.-.- t E re a5 4&t.-.E -iEh to -<.:re r.nt r PM_. Gu e' c f (1942;



























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:u :e j : r u : i pairi r.:r, [.; the rEpr ..(: I ; c C api :.r .hei ra n .

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the tEc.per i..i. i ii,...r., i (he rpperdi* Tl e 2.


Ptr (tc.c M.Pncr ajl frer,T.

A5 :r..:.,.. ;r. Table II per c .c aL t.1,-r.T i cr. .Ma l, ..: re 9. F c ,

1. 5,, j. t. r. t,. .. t.-r t .e :-.itr .i . icie -, returr d r r.. -

r :.ic r. a '* M ;, M" :-: [CF c'.:. :.a c.. r.Ir.I i rr d. lced .-d o. irr adi itEd

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E alu fc-r 1 c .ntr .: *I d t i c, ,id rct.urr.cd jro.,up d.urli .-.q hi .-.

The c .-tr l qr.-.up T .-a' i d per cer.t Ltr,.:.r,,..,l .: r.T,. at .. le e tihrc, hn-

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gr .up begjarn :..- .,i-, ir.crc~ ni l Ir arer cr cer.t .ae :,-' ,'t,:.r.cal pCr,

t,. I li :-.p.. ire .c: c urred ir. the returned .Jr :-u, r;l r'et,-,ce: in percern-

al 4 :. r -.br..;:r .l :1 nT. bel Ec tC [te:0 .r ul: .re rI-.i[ i .j.-. ca.r :[.lt

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As shown in Table II it appeared that treatment with testosterone,

PMS or thyroprotein was not effective In overcoming the harmful effect

of vitamin A deficiency on sperm abnormality in rams, rather it appeared

that hormone treatment had a detrimental effect. Figure 13 illustrates

changes in percentage of abnormal sperm during phase 11 in the hormone

treated and control groups. The A + PMS group had a higher percentage of

abnormal sperm than the control throughout phase 11. The thyroprotein

treated group showed a temporary improvement in sperm abnormality percen-

tages from weeks 3 to 10, from then on an increase in sperm abnormality

was evident. Testosterone treatment produced a gradual decline in sperm

abnormality from weeks 4 to 7. From weeks 7 to 12 values were variable.

After week 12 no further determination of per cent abnormal sperm was pos-

sible because of the reduced sperm concentration present in the semen of

these rams. These phenomena occurred In the A + PMS group from weeks 18

to 21. Differences in per cent abnormal sperm between the returned group

(with comparable values to the control group) and the hormone treated

groups were not of statistical significance. Irradiation appeared to have

increased sperm abnormality. As shown in Table 13 this effect was of sta-

tistical significance during weeks 6, 7 and 8 of phase II. However only

2 rams were present in each of the Irradiated groups at this time. Table

13 also shows that the effects of Irradiation on sperm abnormality were

more drastic during weeks 6 and 8 on the control irradiated rams. Figure

14 illustrates changes in per cent abnormal sperm values for the control

and both irradiated groups throughout phase II.

Analyses of variance for differences between treatments in per cent
abnormal sperm during the first 10 weeks of phase II are shown in the

Appendix Tables 30 through 37.





















II~


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PrEulirt ct-tieJ in this experiment indicate that vitamir A dcrI -

c c.C, ..-crciEE.d (,. presence of *.,Ljr.,,l *pci i in the semen of r.-,,-.n

.II .-I r, h jer,,:,c t diet. This elrr .-:t c.:.ul, J ot be overcome witr. treat-

..ci.t o.r rei t. er.:-.e. PMS or thyroprotein. Figure 15 shows a brca en. r .'

:.ind;i.:,n i' ,cJ rai eri tlc of .per-, r.,.. vitamin A deficient rams. uilti

obtained in this study are i-i, ajree.T'r. with those reported by Lind,'e,

al. (1949) in rams, and Bratton et al. (1948), Erb et al. (19L41 ar.

Gilbert and Hart (1935) in bulls.




As shown in Table II total sperm per ejaculate (xl06) durir. pl.=.e-

I and II were 2945, 1723; 637, 461; 605, 1184; 1627, 1163; 1025, 2.s, 1':.,

313; 3036, 1346; and 1421, 670 for the control, deficient, returrea '

Thyroprotein. A" + PMS, A" + Testosterone, control irradiated and A- .rrj-

diated groups respectively. During phase I total sperm production a.i

somewhat less in rams on the vitamin A deficient diet. The defic-i,,a r,:,,p

had a further drop in sperm production during phase II. A similar Ercr-d

was observed in the controls. However the returned group experie-i:ci 3

recovery during phase II. Figure 16 illustrates changes in total Ipcr.Tr

proOuct l: ':i.n .r [rC cnrol. i i.~ ,cinrt d ri'~urred groups. The alT .r :'Tri-

diate irnr. ;: c r, t :, l :~-: r., per eiac.1 i i, -. the returned group :::urrred

fror, ix.c tr..r.d eek )1. irt *e f,:.i rt ,r :up ra r n-. ed a low spermi pr.-

uCtLl.r. (hr ,u i :,ur[ ih 1 .I ei of pria. II.

A: hou-. ;,r. T,=t. II [hE Pf h : .rj TE:r ): uraC- treated rar: under-

r.nt a3 frcrih r drop I r. perT roduct i.. dl, inrg pr- :e II. The total .per-,.

,cr elcul ate :t ri-.: ini thr. e :? .j e ri..up: .6 ijhuI / lower t-j'. i itre















































F. ,jr. -- :-. I ;.-. .r j..in A deficient rams.






72
















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deficient group, suggesting the possibility of a detrimental effect from

treatment with these two hormones. The thyroprotein group maintained an

average sperm production that was above averages found in any of the groups

on the vitamin A deficient diet. Figure 17 illustrates total sperms per

ejaculate from rams on the hormone treated groups throughout phase II.

The rapid decrease in sperm production in testosterone and PMS treated

rams is clearly shown. The beneficial effect from thyroprotein treatment

on total sperm production was apparent until week 20. Irradiation caused

a drop in total sperms per ejaculate of rams in both groups irradiated.

This effect was more severe in the A" irradiated group. Figure 18 illus-

trates total sperms per ejaculate from rams in the control and irradiated

groups. There was a decrease in sperm production in both irradiated groups

up to week 4. Froms weeks 4 to 7 sperm production in both irradiated

groups was comparable to the control, at this time a decreasing trend began

which remained with the A- irradiated group until termination of the ex-

periment, while the normal irradiated group appeared to have recovered in

week 17.

Differences in total sperms per ejaculate between the various treat-

ment groups were not statistically significant. However some biological dif-

ferences were evident. Vitamin A deficiency had a severe effect on total

sperms per ejaculate. This detrimental effect could not be overcome by

treatment with testosterone or PMS. This supports the findings of Lindley

eta. (1949) and is in disagreement with the results of Gunn et al. (1942).

Thyroprotein treatment on vitamin A deficient rams appeared to have dimi-

nished the harmful effects of the vitamin A deficiency on total sperms per

ejaculate. This has not been reported in the literature. The favorable


























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.a P. DuE C rI:i jc r L, . e.r I ar i Ci, t r- h .I :: I >1 1 1 al Ic : )r.i L ,in L

Occur r. ttE i1 t ,. A -t ii n r -. Tr, et ric t: : r rr r ri .t _t i: :1 :.r ..

prod.J ct ;.:n c 5n Le e l i ir ned i t .r : a :. 5un t l..c ,rr daj 3 i,..,', Ai ecrt d per-

7.E':. c- : ii t. Ir.E arl : t t 3j e t.t :.c ; (r.3 C r ,.. r.e.- :p rma -

C ~c t : re pr. luced *-..'..;al :per pr.:.Jc ht:. .aJ, r : ..i.ed. Tri... -- ld

d* 1i., l I'E rtEC --,. r r, I tr r.T, pr.. u: r i l ,. :... L r e.i ar. ( he t--..r .aI i r r aji e d

gro.,jp 16 ee l, t ie. : r di tat :-r.

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.re Ei. Hir. h .d;* t. -p. o 1T 45..t I i .: r r i r-ce' .-r j li cre.ice

t',.Eu :p -rr.. CF r e ic.l te rd r-,.,,, in, TstIe 3'.1 [5ltr...-j. ,) V.. r.'E

rA -.. ii. p .-I

I t ,P .j5p r ri t Iii p;.0 .-'-E t,:, ra,, :-,eral C o.,cd, 1 : E, r. E["

eC rrcti C, th .e a -.Ti. reat-rc *- cr. :. chtr Lcr, i .: 1 t -.-.re doir.j

l : er.phai ed [r.= r ipr.e.. nt aI ruT.Lter: C re 1 i.i t r. I jr e de a r.

E ; t i-. je the .i'Lntie :r .3 r in te e, :.',ler. tr. -. I ri ,-

L- lit, Ir. t ,j e JEpr ,.i the r.l :. C Al i-.J t c c F c .C : t. r r r ..-

t icoal n; i ic r.ct -. t Ia, t r.i c :eiTen *e j-- A .i_ ,n,.; M e. r

ra,,I .f*a. redu ed_ r ir,;' ei fect h. pr. :r .tr [ ut .,re L e.tern-aT H -,,. r..:r-

ji Ioc it ed TI 'i i h ra-.! i .- e -r, cj it r.t C, ...ere *..t e r r- .:j .n ie -id r it[

ut the r .. T. ; c Er C t jn i-t.E-r. ii:l n i ..i pr t, t rt-J rer.-..:-(










with testosterone, PMS or thyroprotein. Sperm motility was severely

affected by a vitamin A deficiency. This effect did not become evident

until the nervous lesions associated with a vitamin A deficiency began

to show in most of the experimental animals. PMS or testosterone treat-

ment had no effect on preventing changes in sperm motility on the vitamin

A deficient rams. Thyroprotein was of some benefit. Irradiation appeared

to have had a detrimental effect on sperm motility values in both groups

Irradiated. An increase in number of abnormal sperm occurred in vitamin

A deficient and irradiated rams. Hormones used had no beneficial effect

in reducing the percentage of abnormal sperms present. Total sperm produc-

tion was severely affected by the vitamin A deficient condition. Manifes-

tation of this effect occurred in association with a decrease in semen

volume. Of the hormones used only thyroprotein appeared to have had any

beneficial effect on sperm production. Histological examination indicated

that the reduced sperm production was due to a reduction in the germinal

epithelium of the seminiferous tubules which caused an impairment in sper-

matogenesis. The ineffectiveness of PMS or testosterone on improving

semen characteristics would suggest that the effects of a vitamin A defi-

ciency on semen traits do not occur through a secondary effect from im-

balances or reduced secretion of hormones normally associated with sperma-

togenesis.


Histoloqical Investigation

Quantitative evaluation of testes of rams from the various treat-

ments is shown in Table 14.




















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N 0 N 0 0 -











ri liu, cr..:k.-.iss in all rams on the vitamin A deficient diet. The dia-

ic.r *.. r n :eminiferous tubules also appeared reduced. Similar lesions

rha t...r., rtp.rc. d in rats by Mason (1933), Howell et al. (1963), in bulls,

Br:cr.jn j2 (1948) and in rams by Lindley et al. (1949). None of the

hr.:m:ar... ,u.*d nad any effect in preventing testicular damage.

ir. r:critial tissue was not affected by the vitamin A deficient

:j.ji ir., Tt.ere was more interstitial tissue in rams on the deficient

.1ic[ (nr, ir. those on the diet containing vitamin A. The reduced amounts

.:t ;:nrr.riilIl tissue present in the testes of vitamin A supplemented rams

can not be explained. Figures 19 and 20 illustrate the lack of epithelium

thickness in the seminiferous tubules of a vitamin A deficient ram compared

to a control. Note also the sparse interstitial tissue present in the

testes of the control ram.

Thyroid tissues from the control, deficient and A- + Thyroprotein

rams were studied for differences in thickness of the follicular wall cells.

Table 15 shows average epithelium thickness for the 3 treatments. Based

on the small number of thyroid samples studied there appeared to be no

differences in thyroid secretary tissue activity between the 3 treatments.

Histological studies made on the adenohypophisis of rams on the

various treatments revealed no unusual degree of degranulation and vacuo-

lation present. Even though the observations made of the adenohypophisis

were limited in numbers, the lack of abnormal hypophyseal development

suggests no alteration in physiological processes that were dependent upon

the pituitary hormones due to the vitamin A deficiency in these rams.












































Figure 19.--Seminiferous tubules in testes from a control ram.














































Figure 20.--Seminiferous tubules in testes from a vitamin A
deficient ram.












N : (( r :[ r. ,r r "u i-d a iI .* I [ *I ...-. t F .-r r .

I lti i : :r,[ r E.. [rn I r. ... ..r L r,.j I E, I- t I hl I r, v I Ee-.1


S[*C f I ,r. c n r. IT. h I r; .. e.;.Ed i: i t a. i r r

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: :r i:r r..ued pC r..' d . i t ..

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TAuLE li

r.Iuji Tar IVE EL.LUAlT I.iN -.,F THE TiivhmCln [ FACar Pi' IN THE
turt7 L, DcIUtCH'. E rt .Ib Li Li rhilAEe-uP i1I dir.



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Testes were significantly heavier in r --i I.. th. cornticl ir.l re-

turned groups than in all other treatments. Ra..: the "- .rrlral-.J

group had significantly heavier testes than other ra.. : r, Ihe .. i r. ta

deficient diet. This was probably due to the 1fct r-ct -:-t E:.l Ih- rar.:

on this treatment died much earlier in the expel ie.-.n tectaEu: .*1 irra.l.e-

tion exposure thus were subjected to vitamin A cje-.rl. ri ,.. r:r c.-...Or-

able shorter period of time. PMS or testosterone tre.n.li-n. .d.J I'ot preven[

testicular atrophy in vitamin A deficient rams. TI.,r.-: r .. tr tEJ r .,

had slightly heavier testes than rams in the devrcicr.t Jgr.u tut Ihis .ji-

ference was not significant.

Results obtained in this experiment she. it-l.l .i -.-i dotr.c c.

causes a reduction in testicular size. This ei recr d,.,e. ,.:,[ p:,:,r E.. t. r

secondary to a testosterone deficiency since tet:: r :,.i ,.-.i-n : rat. :.,

was not effective in preventing the reduction :- I.:1iCulaI .- c

The reduced testicular weights associate- r ..,r j a- det c..-:c ,

in rams have been reported by Lindley et al. ( .-') ..r..lar re:- r : .-re

reported in rats by Gross (1924), Wolbach (19251 M,,.- anJ Trur.i 1194l

and Howell et a (1963). Mayer and Truant (19'49i .r.recir..-. te:ro .re-

rone were able to obviate testicular atrophy ir. ..ra-r,,i *Jl-t.cenr r ji

Lindley et ai. (1949) were unsuccessful in prec.-,[ ..9 [I:Ciculr cir:.:h;

in rams with injections of testosterone. The re-i c- t r [. : -*.pr,.l-,;

are In agreement with the findings of Lindley Et J. Ii,4,4l.

Epididymal weight comparison between the ar...r.. r-a.T.r,ct 3r:up

is rather inconclusive. Rams in the deficient. A" n: a.-d T-1c, i -

terone groups did not show significantly differe=-r 'e, dd,lii :Ize tr.-r.










the control rams. Rams on the A + Thyroprotein groups showed signifi-

cantly smaller epididymal size than deficient and control rams.

There were no significant differences in seminal vesicle weight

between rams in the different treatments.

The cowpers of rams in the A- + Testosterone were significantly

heavier than those of rams in all other treatments. Rams in both irra-

diated treatments had significantly smaller cowpers than rams in all other

treatments. Vitamin A deficiency appeared to have had no effect on cow-

pers development in rams. The explanation for this irradiation effect is

not known. Testosterone treatment had a stimulating effect on the deve-

lopment of the cowpers. This is to be expected since it has long been

known that the functioning of the cowpers is under testosterone influence

(Turner, 1961).

Statistical differences in adrenal size between the various treat-

ments are not conclusive. However in general rams in the control and re-

turned groups showed smaller adrenals.

A significantly heavier thyroid was present in rams in the A" irra-

diated group. However rams in the control Irradiated group had the smallest

average thyroid size. So that if irradiation per se had an effect on

thyroid development this was only evident in vitamin A deficient rams. No

other effects on thyroid development were observed.

Pituitary weight did not appear to be affected by the vitamin A

status of rams. Treatment with either testosterone or PMS did not appear

to have any definite effect on pituitary development. Had the level of

thyroprotein given been sufficient for normal body functioning, pituitary











Sr..r:.t,i ro r r..:.r r. e :cre[ ld hj.e Let r. reduced CU.-r. uei.C .-[ I a

r Ja .Cl Io .-. t fr.. i i t r r:.- I cL te. rtr .phIc r..r.- r.e : i C .ui t ,,

c.:-ul 3 I I r C ul ted I ir ,i r ere:. irq t rC.IC that Fh ,r.:,rr, I e t r- t -

.-ent did not cause a reduction in thyroid size. However the data are nv:.

conclusive enough to permit a definite statement on this relatior...,-p

Table 25 in the Appendix show individual gland size for .ll ri .:

The reduced number of rams and the variability in gland size bet.-*ee ra-.:

within a treatment suggest caution in making any categoric stateur~c-t: *.

hormonal and nutrrr.onjl interrelationships in these rams.

Analysis of variance for differences in testes, epididyci:. tm--..7a,

vesicles, cowpers, adrenals thyroid and pituitary size are shown i. tie

Appendix Tables 40, 41, 42, 43, 44, 45 and 46 respectively.














SUMMARY


A study was conducted to determine the effects of a vitamin A defi-

ciency on the reproductive capacity of rams. An attempt was made to coun-

teract the harmful effects of the vitamin A deficiency by administering

testosterone, pregnant mare serum (PMS) or thyroprotein. Information was

obtained on the tolerance of vitamin A deprived rams to gamma irradiation.

The experiment was started on February 2, 1963. During phase I of

the experiment 12 yearling rams were assigned as controls to a purified

diet containing 3,000 I.U. of vitamin A per Ib. of feed. Thirty-six year-

ling rams were placed on the vitamin A free diet. During phase II the 12

rams on the control diet with average vitamin A values of 139 mcg./100 ml.

plasma and 156 mcg./g. fresh liver were continued on the same ration with

6 serving as controls and 6 irradiated with 400 r of whole body gamma irra-

diation. During phase II the 36 rams on the vitamin A free diet with

vitamin A values averaging 5 mcg./100 ml. plasma and 5.6 mcg./g.fresh liver

were continued on the vitamin A free diet and were assigned in equal groups

to one of 6 treatments: (1) 400 r of whole body gamma irradiation; (2)

Daily injections of 60 RU of PMS/ 100 Ibs. body weight; (3) Daily injections

of 10 mg. of testosterone/100 Ibs. body weight; (4) Daily capsules of 1 g.

thyroprotein/100 Ibs. body weight; (5) 3000 I.U. of vitamin A/lb. of feed;

(6) No additional treatment. Semen was checked monthly during phase I

and weekly for 21 weeks during phase II.

Night blindness, cloudiness of the cornea, uncoordinated gait and

extreme weakness were symptoms associated with rams on the vitamin A free













d i. L.T.ir d i.-,I.[ rr, l t,.c,,,:.rrl..~ -..i n I he Cc:[ Ca I ".~ cr.ar r cr t

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:.:,n , le c Jd r,,. i 1, r ',rr . 91'"( rh ,r. C 1 ro l L'ur -..-) l, a ie II

PL :.r c t .. 1e' I r.c re c .Tc.' d nd, ri e an, t. i.cv C l I I cCi *r. c r f e-

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i. ei re atlc I. :, rdC. er J'Jd utrr.rt ed n: j.[ hi I : t .ri C UUU I .1.

L i .. a-,T.,, 1 C i rC Z .i C e Ir.....di : r I t [c c r ar, .n,,:,c ra ..;..

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.it ,- .-i (" :. r,.i -* L i Ed [:. ,rrad jil *. j 1 .1or[ r iF CC the r .ur -

I. l [. c ( r. dc I d e..C Cd a c -J 3 F 13r eCuiF ,r., .- ar..n l n p C Ere -

'U. Ihi . j h i C.-. [he JJ.d lec I .. rh S.r.l- r- c t li i .E I ( e:[<

H. :[ul.A iC al rai ce- .:, 1 [ic .r,,r,..J ,'..1 ade...i-.,h, ,. i-, .. : : r al ,.i ,.;. Ch...,) i

lihd C:.:ul Lc ail Lult I t. .. [ m.n 4 r .:.r r .T: de.' a Iu.' i:.er.

I,:.( .I I per r i jl 'r -r.7".al "pD r.. r.,I 1 31 f. ri pr -JuC I ,...n r re l,

'tcCr cd t, .t a.T.i r. .J c C e-r. .per.i r_. I l. r, a-,d [(.(tal 'per' pcr., J.,J

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pc r CE I 3-r reduCc d *.:. 1 al uI . r .;..: i n.: r '3 i . aT.. i








91


reserves. Testicular weight was reduced in all vitamin A deficient rams

Epididymis cowper's, seminal vesicles, adrenals, thyroid and pituitary

weights were not affected by the vitamin A deficiency.




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