THE EFFECT? OF ENVIRONMENTAL TEMPERATURE AND LIGHT GEN THYRGID ACTIVITY AND CERTAIN METABGLEC MEASURES IN SHEEP Thesis Ior IIIO Degree oI DIN. D. MICHIGAN STATE UNIVERSITY Hugh Edward Henderson 1958 lfit‘blfi This is to certify that the thesis entitled THE EFFECT OF EIG‘.’IROIII~ENTAL TISI’PERATURE AND LIGHT ON THYROID ACTIVITY AND CERTAIN I‘ETABOLIC I'vIEx‘sSURES IN SIEEP presented by Hugh Edward Henderson has been accepted towards fulfillment of the requirements for Ph. D. degree in Arlimal. Hquandry Wm Major professor Date 5/20/58 0-169 LIBRARY Michigan State University THE EFFECT OF BIN IRONITSNTAL Tszcasmrms mu) LIGHT ON TI—I‘fliOID ACTIVITY AND CERTAIN METABOLIC MEASUKES IN SHEQP by Hugh Edward Henderson AN ABSTRACT Submitted to the School for Advanced Graduate Studies of Michigan.State University of Agriculture and Applied Science in partial fulfillment of the requirements for the degree of Doctor of PhilOSOphy Department of Animal Husbandry 7 1958 Approved \_,7 .Zégi/r:2;2::/7Lékn4AZwL// ABSTRACT THE EFFECT OF WRONMENTAL TmPEEATURE AND LIGHT ON THTROID ACTIVITY AND CERTm METABOLIC MEASURES m SHEEP Hugh E. Henderson The effect of ambient temperature and artificial light on thyroid activity and other metabolic measures in ewes was conducted from June 18, 1957 to January 18, 1958. A total of 12 experiments involving seven ewes each was conducted in controlled temperature and light chambers. Each experiment was con- ducted over a period of 27 days with seven days devoted to acclimatiza- tion and 20 days to collection of data. All experimental animals were maintained in a similar environment prior to being placed on experiment and self-fed feed and water prior to and during the experimental.period. Estimates of thyroid activity for each temperature and light con- dition studied were obtained by thyroidal uptake or retention of 1131, output half-time of 1131, l-thyrqxine secretion rate, chemical analysis of thyroidal 1127, and histological determination of thyroid epithelial cell height. Estimates of thyroid activity Obtained by all five methods were in very close agreement with the exception of thyroidal content of 1127 and percent uptake of 1131 which failed to reveal any significant difference among the various temperatures studied. It was demonstrated that ambient temperature had a profound effect on thyroid activity, with high temperature greatly suppressing and low temperature stimulating thyroid activity. -2- Hugh E. Henderson Daily'body weight gains, daily feed intake per pound of body weight, daily'water intake, rectal body temperature, and respiration rate were all found to be significantly affected by ambient temperature. Ewes subjected to 12 hours of artificial light daily showed signi- ficantly greater values for thyroid activity than did ewes at 8 or l6 hours of light as measured by output half time. Daily hours of artificial light had a significant effect on feed consumption and respiration rate, but had little or no effect on rectal ”body temperature, water intake, and body weight gains. From the correlation analysis, a highly significant inverse relation- ship was observed between maximum uptake or retention of 1131 at three, five or ten days after injection and output half time. These correlations demonstrated that an actively secreting thyroid was accompanied by a low uptake or retention of 1131 and vice versa. Retention values at ten days post injection of 1131 were more reliable estimates of thyroid ac- tivity than three day uptakes and appeared to be a reliable inverse estimate of thyroid activity in ewes at temperatures below 90° F. Th; JFFECT OF ENVIRONMJNTAL TEMPERATURE AND LIGHT ON THYROID ACTIVITY AND CERTAIN METABOLIC MEASURES IN SHEEP by Hugh Edward Henderson A THESIS Submitted to the School for Advanced Graduate Studies of Michigan State University of Agriculture and Applied Science in partial fulfillment of the requirements for the degree of Doctor of PhiloSOphy Department of Animal Husbandry 1958 Hugh Edward Henderson candidate for the degree of Doctor of PhiIOSOphy Final examination: May 13, 1958, 1:30 p.m., 103 Anthony Hall Dissertation: The Lffect of Anvironmental Temperature and Light on Thyroid Activity and Certain metabolic measures in Sheep. Outline of Studies Hajor subject: Animal Husbandry Einor subjects: bhysiology Anatomy - Pathology BiographICal Items Born, September 2, 1923, Glover, Virginia Undergraduate Studies, Virginia Polytechnic Institute, l9hl-l9ho Graduate Studies, Vi ginia Polytechnic Institute, 1953-1956 ifichigan State University, 1956-1958 Experience: Member United States Navy, l9h3-19h5, Farm Manager l9h6 Assistant County Agent, l9ho-l9h8, County Agent 19h8-l9Sl, Assistant Professor, Virginia Polytechnic Institute, 1952, _ Associate Professor, Virginia Polytechnic Institute, 1953-1950, Graduate fiesistant, hichigan State University, 1950-1958. Member of Smerican Society of Animal Production, Royal Order of Squanto, Sigma Epsilon, and Sigma Xi. ACKhChLflD-dHDhTS The author wishes to express his deepest appreciation to Dr. Earold A. Henneman, under whose guidance and supervision this work was done, for his interest, encouragement, inspiration, and help throughout the course of study. Sincere thanks and acknowledgement go to Dr. S. P. Reineke for his constant interest, guidance, and supervision during the study. His ever willingness to offer assistance and inspiration will always be remembered. Grateful appreciation is extended to Dr. William T. Rages for his assistance in the design and statistical analysis. Sincere thanks are expressed to Dr. C. K. thitehair and Dr. Lois Calhoun, other members of the guidance committee, for their willing assistance, advise and encour- agement. The opportunity to have worked with the entire committee and known them as a scientist, scholar, teacher and friend will always be cherished. Sincere thanks are extended to Lee Bell, shepherd, for his assist- ance in caring for experimental animals and Hrs. Kay Butcher for an excellent job of typing and arranging the manuscript. The author further thanks and is grateful to hichigan State Univer- sity for financial assistance in the form of an assistantship and making experimental facilities available for the study. r1 ‘1"; '_1 "‘1 ‘v :1 1", r1 .Lulng 01‘ C‘vi 1.41.1‘8 “—Vf'a -.. ’1‘.., .I..1-J.-.'J-/I-C.L.1.Uil‘. o o o o o o o o o o o o o o o o o o o 0 ma“ nnTr .1, LMUJJU1J. ..JL.) 0 O o o o o o o I o O o O O o e o o a O o .) vrl . ‘)l I. l‘ "J- .’ " A...“ .. IA. ..1 1.41c~~1 v-11.) o o o o o o o o o o o o o o o o " "1" ~ ‘3‘ ’\ 7" '"“ .' 1;.JII:-'1)LJ .11.-=J 1-‘.I.‘~»C._J..n).L.J o o o o o o o o o o o o o o o . V" 91:. _ 4",:‘4y't'7 ‘ .‘ :- .‘j' LLAVULLQ -LL~J.J a—J‘u—L-Lu/klkjdk/d—Vl‘ . o O Q g o Q Q Q g Q g g g . L"*‘L 9 ffect of “mbient Temperature and artificial ‘ ' 1‘“ 'V n — Q Lifnt on Ihyrcinal Uptake OI 1121. . . . . . . . B. Lffect of anticnt Temperature and Artificial Light on Thyroidal Output Turnover Hate of I 31. C. Lffect of gmbient Tenn rature and Artificial Light on Secretion Rate of l-Thyroxine . . . . . U. E“fect of Ambient Temperature and Artificial Li; ht on .c17“: YIII. BLLJJLOJ1LALIIII o o o o o o o o o o o o o o T A'DT) 5‘7": v" VILI."L.1..J1~L'.L){OOOOOOOOOOOOOOOO \O 10 12 115T OF TABLJS One Day Percent Uptake of 1131 by the Thyroid Gland in iwes Under Controlled Ambient Temperature and Artificial Light Conditions . . . . . . . . . . . Three Bay Eercent Uptake of 1131 by the Thyroid Gland in Ewes Under Controlled Ambient Temperature and -LrtifiCifll Jii'i’lt Conditions 0 o o o o o o o o o o neximum Percent Uptake of 1131 by the Thyroid Gland in Lwes Under Controlled Ambient Temperature and arti- ficial Light Conditions. . . . . . . . . . . . . . Zero Tine rercent Uptake of 1131 by the Th; in dwes Under Controlled Ambient Temperature and Artificial Light Conditions. . . . . . . . . . . . bays of gaximum Percent Uptake of 1131 by the Thyroid Gland in “wee Under Controlled Ambient Temperature and Artificial Ligo Conditions. . . . . . . . . . . . E fects of Ambient Temperature on Turnover date One- rroid Gland half Time of Ii3l by the Thyroid in Ewes . . . . . . . Effects of nrtificial Lifht on Eurnover Rate Une- half Time of 11:1 by the Thyroid in ores . . . . . . . fistimated naily l-Thyroxine Secretion date of Ewes at Controlled ambient Temperature and irtificial Light conditions 0 O O O I O I O O O O O O O O O O O O 0 Effect of Ambient Temperature and artificial Light on Height and Iodine Content of Thyroid Gland . . . . lffect of Ambient Tenperature and Artificial Lifht on Spithelial Cell Heifht of Thyroid Gland. . . . . . Daily Body height Gains of dues Fed ad. lib. Under Controlled Ambient Temperature and Artificial light condi ‘ti ODS O O O O O O O O O O O O O D O O O O I 0 Daily Eeed Consumption Per Pound of Lody'fieight of owes Fed ad. lib. Under Confrollcd Ambient Temperature and —“‘$I‘tiiiCial Li‘jfl’b corlditionSo o o o o o o o o o o 0 2h 25 26 27 31 37 no L2 15 16 l7 l9 LIST c? TisT 5 (continued) rounds or later Consumed Laily by lxes tnder Controlled inbient Ternerature and Artificial Lifih Conditions. . . “ffect of Controlled gmbient Tenperature and irtificial Lifht Conditions on Rectal Lody Temperatures in ewes . . nffects of inbicnt Temperature and Artificial nijht Con- ditions on hesniration rate in dues (flank movements per minute ) O O O O O O O O O O O O O O O O O O O O O O O O 0 Summary of Certain PhysiolOQical desponses to Controlled imbien Temperature in Qwes. . . . . . . . . . . . . . . Summary of Certain Physiological Responses to Controlled Ih'tifiCial Lirht in. Jules o o c o o o o o o o o o o o o 0 Correlation between Controlled Ambient Tcnnerature and Certain.khysiolOfical desponses in gees. . . . . . . . . Correlation ietween Turnover date One-half Time and Certain EhysiolO'ical desponses in ewes. . . . . . . . . \_ ’1. CL) LIST OF FIGURES Typical Uptake—Cutout Curve of 1131 by the Thyroid Glarld Of £31180? 0 o o o o o o o o o o o o o o o o o o o o 30 Haximum Percent Uptake and Turnover Late of 1131 by the Thyroid Gland in Ewes Under Controlled ambient Temera-L—Im‘e O C O O O C O C C O O O O C C O O C . . O . C 32 Maximum Percent Uptake and Turnover Rate of I131 by the Thyroid Gland in ewes Under Controlled Artificial Light 0 O O O O O O O O O O O O O O O I C O O O O O O O 0 3b U L—I Hi b—‘I LIST Ob APfiQLQICJS Turnover date of I131 by the Thyroid in hues at 500 r. Controlled Ambient Temperature . . . . . . . . . . . . Turnover Rate of 1131 by the Thyroid in Lees at 550 E. Controlled Ambient Temperature . . . . . . . . . . . . Turnover hate of 1131 by the Thyroid in Jwes at 850 B. Controlled Ambient Temperature . . . . . . . . . . . . Turnover Rate of 1131 by the Thyroid in fives at 900 L. Controllei Ambient Temperature . . . . . . . . . . . . Turnover date of 1131 by the Thyroid in Ewes at 8 H0138 COHtI‘OllCd —rtifiCiC‘Ll Ligl-lt o o o o o c o o o o Turnover date of 1131 by the Thyroid in fines at 12 Hours Controlled Artificial Liyht . . . . . . . . . . Turnover date of 1131 by the Thyroid in Lees at 16 Hours COfltrOlled ArtifiCial Li:ht o o o o o o o o o o Estimated'Daily l-Thyroxine Secretion Rate of Lwes at Controlled Ambient Temperature and Artificial Light conditionSooooooooocoooooooooooo One Day'Percent Uptake of 1131 by the Thyroid Gland of Ewes Under Controlled Ambient Temperature and Arti- fiCial Light Conditions. 0 o o o o o o o o o o o o o 0 Three haw Percent Uptake of 1131 by the Thyroid Gland of Ewes Under Controlled Ambient Temperature and Arti- ficial Light Condition . . . . . . . . . . . . . . . . haximum Percent Uptake of 1131 by the Thyroid Gland of Ewes Under Controlled Ambient Temperature and artificial Lj_E~h-t Conditions 0 O O O O O O O O O O O O O O O O O O ' 10 V ' "' fl ( ' ‘ W ‘0 Zero line fiercent Uptake of lljl by the Therid Gland of Ewes Under Controlled imbient Temperature and arti- fiCiaJ. Li;ht Conditions. 0 o o o o o o o I o o o o o o F‘IL Day of Maximum Tercent Uptake of 1131 by the lnyroid F'fl Gland of Lwes Under Controlled Ambient rennerature and :‘mtifiCial Li‘ht Conditions. 0 o o o o o o a o o o o 0 7h 75 7o 77 78 79 80 LIST or iii": _;r.;;lC’jS (continued) 7“?" ‘ I'IY-I—_ ‘ Ari" .' ‘11“! .' M w a: £1.) (“Q G) N Twenty Lay Cody height Gains of Ewes Fed ad. lib. Under Controlled Ambient Temperature and artificial Lifiht Conditions. . . . . . . . . . . . . . . . . . . do O Twenty Bay Feed Consumption Ter Pound of Body height of Lees Ted ad. lib. Under Controlled Ambient Temper- ature and Artificial Light Conditions. . . . . . . . . 87 P Pounds of later Consumption Daily'by Cwes Under Controlled Ambient Temperature and Artificial Light conditionSooooooococoooooooooooo 8'8 Q Rectal Eody Temperature of fives Under Controlled lmbient Temperature and Artificial Liiht Conditions. . C? R Respiration hate of Lwes Under Controlled Ambient Temperature and Artificial Light Conditions (Blank movements per minute). . . . . . . . . . . . . . . . . 90 S The Determination of Iodine in Thyroid Glands. . . . . 91 T Cffect of Ambient Temperature and Artificial Light on height and iodine Content of Thyroid Gland . . . . . . 9h U Lffect of Ambient Tennerature and Artificial Light on Epithelial Cell height of Thyroid Gland . . . . . . 95 V Testing Lifierence Between TLO Correlation Coefficients 9o I. IhTRDQU TICK It is well established that both environmental temperature and the length of davlight has a pronounced effect on the physiology of prac- tically all of the life processes important to meat animal production. Numerous experiments indicate that both the length of day and ambient temperature play a major role in the seasonal breeding habit of the ewe, and surfest that its action mav'be exerted through the pitu- itary-thyroid gland relationship which controls body metabolism. Since previous research indicates that thyroid activity in the ewe varies from one season of the year to another, it is necessary that re- liable estimates of thyroid activity be made at various controlled ambi- ent temperature and light conditions in order to intellirently adjust activity estimates made at various seasons in tie year for comparative purpos s. Furthermore, if thyroid therapy is a factor in overcoming summer sterility in the ram and anestrus in the ewe, it is necessary to have reliable estimates of normal thyroid activity at the various environ- mental temperature ard light conditions. This investifation has been designed in order to gain more basic in- formation on the normal thyroid activity of sheep under controlled am- bient temperature and light conditions and the interrelationship of thyroid activity to certain metabolic activities in the ewe. hith a clearer understanding of the effect of ambient temperature and length of dayliyht on thyroid activity, a more intelligent approach can be made toward extending the breedine season of sheep and a clearer appraisal made of the effects of thyroid activity on reproductive per- formance and producing ability of the ewe. II. OBJECTIVES 1. To determine the effect of controlled ambient temperature and artificial light conditions in ewes on thyroid activity as measured by: a. Uptake of 1131 b. Turnover rate of I131 c. l-Thyroxine secretion rate d. Thyroid content of 1127 e. Thyroid epithelial cell height 2. To determine the effect of controlled ambient temperature and artificial light conditions in ewes on: a. Body weight gains b. Feed consumption 0. Water consumption d. Respiration rate 8. Rectal body temperature 3. To correlate ambient temperature and artificial light conditions in ewes on certain metabolic and physiolOgical functions. 4,- III. REVIEW OF LITERATURE Following the classical work of magnus-chy (1395) who observe: that the feeding of thyroidal materials caused an increase in the oxygen - carbon dioxide exchange, it has been well established by num- erous investigators that the thyroid hormone regulates the rate of metabolic exchange in mammals. Most of the early estimates of thyroid activity were based on the measurement of carbon dioxide by respiro- meters as reported by Hdrch (1929) and Teitelbaum and Horne (l9hl). Many investigations have been conducted to determine the role that the thyroid hormone plays in the various life processes and in partic- ular, growth and reproduction. Furthermore, the effects of environ- ment on thyroid function and its interrelationship to other endocrine functions have received the attention of many investigators in recent years. The role of thyroid hormone in reproductive physiology of the fe- male has been reviewed quite completely by Reineke and Soliman (1953). They concluded that there is a reciprocal balance between the hormones of the pituitary, the ovary, and the thyroid. Through the influence of estrOgen on pituitary thyrotrOphin, the thyroid undergoes rhythmic fluctuations in secretion rate that in turn regulate the output of gonadotrophins and also modify their action on the ovary. Bogart and hayer (l9h6) concluded that the thyroid gland appears intimately concerned with spermatogenic activity of testes in sheep and that the failure of thyroxine to affect semen volume, or motility ratings in rams with lowered fertility during the period of high summer temperature indicates that the thyroid affects the spermatogenic tissue of the testes and has little or no effect upon the interstitial tissue or the accessory organs which are dependent on androgens. Maqsood (1950) reported that mild hyperthyroidism stimulated sperm- atOgencsis and increased secretory activity of interstitial tissue as judged by growth and activity of accessory organs and sexual behavior of growing mice, rabbits and rams. Thyroidectory and prolonged thiouracil feeding interferred with.spermatogenesis and increased fib- rinous tissue in the interstitial spaces. Jeep (1933), Martinez Campos (19h7), and Wheeler and Hoffman (l9h8), working with domestic fowl and Meites and Chandrashaker (l9h8), working with rats and mice, reported that milk hyperthyroidism increased sperme atogenesis. Organon (1936) cited evidence that thyroid therapy was effective in some cases of human male and female sterility. Petersen.gt; El; (l9hl) found that the administration of thyro- protein to hypothyroid cattle improved both sexual libido and fertility. Masson (19h?) showed that the response of the accessory sex glands in castrated male mice to testosterone was increased by thyroxine administration. Black gt:_al; (1950) reported that the addition of testosterone and therprotein increased semen volume in rams during the summer months. 'Warwick‘§£:.glé (l9h8) concluded that during the summer months semen produced by rams receiving low levels of thyroprotein was slightly superior to controls. Smelser (1939) reported that thyroidectomy and administration of excessive quantities of thyroxine resulted in functional reproductive abnormalities. Oloufa (1951) reported that high ambient temperature greatly reduced semen volume, motility, sperm concentration, and percent normal sperm in rabbits and that continuous heat had a more pronounced effect than intermittent heat. The addition of thyrotrOphin under continuous heat caused a further reduction of fertility but under intermittent heat, it improved fertility. Berliner and warbritton (1937) placed rams in a hot room during the winter period and brought about a stoppage of spermatogenesis. Rams which were thyroidectomized were brought back to fertility by injections of thyroxine. Fertility of rams with intact thyroid glands was greatly improved by injections of thyroxine. Biological assay of the pitui- taries showed that rams with.poor semen quality were also low in thyro- traphic hormone. Reineke §32_§l:_(19bl) reported that following thyroidectomy'of goats, there was a reduced content of pituitary gonadotrophins. VanDyke and Chen (1933) found that hypothyroidism in rabbits causes a reduced pituitary content of gonadotrophins which was also true for animals thyroidectomized. ‘ Numerous investigations have clearly indicated that thyroid func- tion was markedly auénented during exposure of mammals to reduced environ— mental temperature and retarded by elevated environmental temperature; however, little data has been reported on the quantitative thyroid secretion rate of mammals at controlled high and low environmental temper- atures. Some of the earliest evidence of increased thyroid activity at low environmental temperatures was reported by Cramer (1916), seidell and Fenger (1912), kills (1918), and hendall and Simonsen (1928). Dempsey and astwood (l9h3) observed that thyroid secretion rate in rats increased markedly at low environmental temperatures and was depres— sed at high temperatures. i-iixner _€_3_’_C_:_ 31; (191111) reported that the thyroid of the chick res- ponded in a similar manner and also deineke and Turner (l9h5) observed a seasonal trend in the thyroid secretion rate of young chicks. A.pronounced decline in thyroid secretion rates of mice were re- ported by'Hurst and Turner (l9h8) when the temperature was increased from 80° to 87° F. Turner (19h3) reported a seasonal trend in thyroid secretion rate in hens and a pronounced decline in the secretion rate of older hens. Kaqsood and deineke (1950) observed a ten-fold reduction in toler- ance of mice to therprotein feeding when the environmental temperature was raised from 2&0 to 30° 0. They suggested that tolerance limits for thyroid stimulation were determined by the temperature regulating capacity. Half-time turnover rates of extrathyroidal 1131 thyroxine were found to averaée 25 hours in shorn and 38 hours in unshorn sheep by Freinkel and Lewis (1957) when both groups were exposed to winter temperatures in Great Britain. Henneman.§§:_§lz (1955) reported thyroid secretion rates in ewes to be lowest during summer months and highest during winter months and that pregnancy had little effect on secretion rate. Since the advent of radioactive iodine (1131) much research has been conducted in order to improve estimates of thyroid activity in mammals and more recently, in the live intact animal. Perry (1951) measured the rate of loss of 1131 from the hyroid gland of the rat with an external counter and determined an index of the secretion of thyroid hormone. By checking the validity of the index against the effect of thyrotrOphic hormone, goitrogens and thyroxine on the rate of loss of 1131, a relationship was established between dose of thyroxine and degree of inhibition of thyroid hormone secretion. Henneman gt; a1; (1955) measured thyroid secretion rate in ewes by injecting graded dosages of l-thyroxine following maximum uptake of 1131. Counts were taken at the end of each 3 day l-thyroxine injec- tion period and percentage of previous count compute . Using percentage of previous count and level of thyroxine injected, a straight line was computed by regression analysis. The point at which the line crossed 100 percent of previous count was assumed to be the secretion rate of the ewe in question. Reineke and Singh (1955) using essentially the same technique as that outlined by Henneman, were able to determine the thyroid secretion rate in rats. Values compared very closely with those obtained by the goitrOgen technique. Using the same technique, thyroid secretion rates have been deter- mined for dairy cattle by Lewis _e_t_._ Ell: (1955) and Premachandra _e_t_,_ 31; (1957), and for goats by Flamboe (1958). Percent uptake of 1131 by the thyroid gland as an indicator of thyroid secretion rate is a debated point by many investigators at the present time. Reineke 33, 31:. (1956) and Chai §_t_,_ gr, (1957), in studying 1:131 uptake and output rates in inbred mice, found a reciprocal relationship between percentage uptake and output rate; a high uptake was accompanied by*a low output rate and vice versa. They theorized that the low uptake - high output rate relationship meant that there was either a larger thyroidal iodine pool prior to injection or that the rate of uptake was more rapidly overtaken by the rate of discharge in the rapid output rate group. The data collected in this study agrees with that of Reineke and Chai; however, frequent reference has been made in the literature that a high uptake of 1131 was positively associated with a rapid turnover rate and a low uptake was associated with a slow output. The effect of ambient temperature on physiological responses in mammals has received the attention of many investigators in recent -10.. years; however, most of the studies in the ewe and ram have been in relation to reproductive performance. McKenzie and Phillips (1933) concluded that temperature seemed to cause no significant difference in the length of time required for ewes to come into estrus in a trial covering a ten day period of time in Lugust with ne group under natural conditions and the other rroup confined to an iced cellar. Yeates (1953) concluded that high temperature was apparently'with— out effect on the incidence of estrus in ewes but was detrimental to satisfactory gestation. Lhen ewes were treated with 7 hours of 1070 F. heat daily from breeding until parturition, there was a fifty percent reduction in number of lambs born over the controls which received no heat treatment. Butt and B sh (1955) observed that ewes placed in a cold chamber at h80 F. with natural light on Kay 26 came into estrus hd day later on the average, as compared to ewes in a hot chamber at 88° F. which came into estrus 9h days later. Rectal body temperatures for the two groups were 102.00 F. and 103.110 F. with respiration rates of 28/m and 150/m in the cold and hot chambers respectively. Rams in the cold chamber averaged 1.9 services per ewe conception as compared to 5.3 services per ewe conception for the hot chamber rams. Dutt gt,'al,_(l956) concluded that fertility and normalcy of ova in ewes exposed to elevated environmental temperatures were nore severely affected than survival of the young embryo. shearing the ewe before exposure to the elevated temperature modified the detrimental effects. Green (19h0) observed that semen quality in rams decreased from January to Pay, June and July ejaculates were quite inferior, July to October showed a rapid increase toward high quality, and the highest quality was obtained from October to January. Foote gt; 3;; (1957) observed that at high ambient temperatures, testes temperature of rams increased at a faster rate than rectal temperature. Lee (l9h5) observed a very low rate of lay in laying hens at high \ environmental temperatures and concluded that 80° r. appeared to be the critical temperature. Casady (1953) observed that the duration of exposure to high temper- atures in dairy bulls had a tremendous effect on fertility levels and that subjection of bulls to 850 to 900 F. for five weeks may interfere with spermatogenesis, reduce sperm concentration and motility and increase the number of abnormal sperm similar to effects noted during exposure to 1000 F. for two weeks. Casady'gtgialL (1956) in experiments with young dairy bulls at chamber temperatures of 60° to 95° F. observed that correlations between rectal temperature and respiratory rate, rump skin temperature and scrotal skin temperature were lower than the correlation between cham- bem temperature and the same responses. On the other hand, water - 12 _ consumption was more closely related to rectal temperature than chamber temperature. Field studies of Johnston and hranton (1952) indicated significant correlations between body temperature and ambient temperature only when the ambient temperature was within hOO to 600 F. and 700 to 800 F. intervals. despiration rate was significantly correlated with ambient temperature and humidity, except when maximum temperatures were within the 80° to 850 F. and 850 to 90° F. intervals. Pulse rate was signi- ficantly correlated with mean daily temperature within the 85° to 900 F. interval. Later investigations by Johnston and nranton (1953) and Johnston 23;.Eia (lSSh) showed that only respiration rate was consistently related to climatic conditions, with absolute humidity and mean environ- mental teuperature being most highly correlated with respiratory responses. McDowell pt; 31; (1953) with Chamber studies on respiratory activity as an index of heat tolerance concluded that respiration rate was not significantly correlated with rectal temperature under the conditions f heir experiment. Similar conclusions were drawn by i and Findlay (1955a, 1955b) from results of chamber studies with bulls. Fletcher and Reid (1953) in studying heat tolerance of feeder lambs on pasture in Mississippi, found that shorn lambs showed rectal temper- atures of 10h.6° F. as compared with unshorn lambs of 105.h° F. Miller and nonfe (19h6) observed that body temperature and respira- tion rate in sheep were very hivhly correlated with ambient temperature. - 13 - In experiments with swine subjected to chamber temperatures from hOO to 1150 F., Heitman and Hughes (19h9) observed that as air temper- ature increased, rectal temperature and respiration increased and pulse rate decreased. Feed consumption decreased as air temperature increased. Under their experimental conditions, rate of gain was great- est and the amount of feed required to produce 100 pounds of gain was least at an average temperature of approximately 750 F. for hogs weigh- ing 70 to lhh pounds and approximately 600 F. for hogs weighing 166 to 200 pounds. As the air temperature was increased or decreased above or below these averages, rate of gain declined and utilization of feed was lowered. At 96° F. a rise in relative humidity from 30 to 9h percent produced rapid distress in hors weighing over 200 pounds, and the respira- tion rate and body temperature increased rapidly. Heitman §t=_al; (1951), in a later study with pregnant sows, found that increasing ambient temperature markedly elevated respiratory rate and rectal temperature. Feed and water consumption were greatly lowered at temperatures above 950 F. No evidence was accumulated that indicated that rise in body temperaoure under these conditions would cause abortion of a normal litter; however, Ragsdale e}; a}; (1919) reported that two dairy cows aborted h.5 and 6 month fetuses when exposed for 27 hours to a temperature of 100° F. Ragsdale §£=.§l& (1953) presented tabular and graphic data on milk production and composition, feed and.water consumption, and body weight in Holstein, Brown Swiss, Jersey and Brahman cows at low and high rela- tive humidities at various temperatures. They showed that below 75° F. atmospheric temperature, the effect of relative humidity on the above processes is not sirnificant. The effect of humidity increases with increasing temperature above 750 F. Robinson and Lee (19h?) observed from experiments with hens, sows, and ewes on high and low planes of nutrition that under hot conditions, the reactions of animals on the high plane of nutrition were signifi- cantly greater than animals on a low plane of nutrition. In another series of experiments where animals were fed rations varying in protein from 5 to 28.5 percent and exposed to a critically hot atmosphere for seven hours, in no case was any evidence obtained that a high prOpor- tion of protein had any significant effect upon the reactions of animals to heat. Investigations on the effect of length of day or hours of artificial light on physiological responses in mammals have been devoted almost exclusively to reproduction aspects with little or no data on growth rate, feed and water consumption, body temperature, and respiration rate. Bissonnette (1932), harshall (19h0), and hart (1950), all working with ferrets, observed that the time of onset of the breeding season varied with the intensity of light and that the frequency of 1iyht to dark was the important factor rather than the amount. The ratio of 2 hours of light to 1 hour of darkness seemed to be about the desired ratio and a reverse ratio caused the animals to go into anestrus. Hammond (l9hh) pointed out that the breeding season in sheep normally occurs at about the shortest day of the year and that ewes bred the -15.. first half of the season had more twin lambs than those bred in the last half. Sykes and Cole (l9hh) observed that when the plane of light was increased daily until 3 hours were added in Larch, and then decreased light 1 hour per week until Lay, five of eight ewes were bred and lambed five months previous to the normal time. Yeates (19h?) concluded that temperature had no effect on bringing ewes into estrus; whereas, decreasing the plane of light in the spring brought about estrus and increasing the plane of light in the fall pre- vented estrus. He concluded that estrus occurred 1h to lo weeks after ewes were put on decreasina 1iyht and estrus ceases 1h to lo weeks after increasing the plane of livht. Hart (1950) concluded that a gradual decrease of lifht mas not necessary to brinc about estrus in ewes but required only a ratio of 1 part light to 2 parts dark. hafez (1950), (1951), (1952) observed that continuous liyht as well as 1o hours of light and 8 hours of darkness broufht about anestrus in ewes. Eight hours of 1i ht and 16 hours of darkness was effective in bringing ewes into estrus. Mercier and Salisbury (19h7a) studied the records of 125,000 cows artificially bred to 71 bulls and observed that fertility of bulls was highly correlated with length of daylight. One to two months of in- creasing light was necessary before the effect of dayli ht reached its maximum. _ 16 - hercier and balisbury (19h7b) also observed a significant correla- tion between hours of daylight and fertility level of bulls as measured by services per conception. Lowest fertility was observed in winter and spring, and hithest in summer and fall. - 17 _ IV. KLTHDJS ALL PAOCSJURE In order to control ambient temperature and light conditions, two temperature and lihht control chambers were used in this study. Animals were confined to 3 by 6 foot indi'idual pens within the chambers in order to obtain individual feed and water consumption records. The cold chamber consisted of a reconditioned meat cooler measur— ing 10 feet wide, 20 feet long and 12 feet high. The chamber was cooled by recirculating air over cooling coils with electric blowers. For each of the experiments, temperature was maintained within plus or minus one defree of the temperature reported. Sven with the aid of a dehumidifier, it was imaossible to reduce the rela ive humidity .- 1' in the cold chamber to that of the hot chamber. a range of 70 to 8h percent relative humidity was maintained in the cold chamber. Ragsdale EE:.§l;.(l953) reported that high relative humidity did not measurably affect metabolic functions in dairy cattle unless ambient temperature was in excess of 75° F. It was assumed that the high humidity in the cold chamber did not alter the experimental results. All natural daylight was excluded from the chamaer and artificial light was provided by four 300 watt light bulbs suspended 5 feet above the floor. An electric time switch controlled the number of hours the lights were on during each 2h hour period. fhe reported hours of arti- ficial light in all cases was a continuous amount of time out of each 2h hour period. -15- The hot Chamber was of masonry construction and measured 20 feet wide, 20 feet long and lo feet hirh. The chamber was heated by recir- culating air over steam coils with an electric blower and temperature was maintained within plus or minus one degree of the reported temper- ature. helative humidity varied from 60 to 70 percent throurhout the experimental period and a dehumidifier was not used. all natural light was excluded from the chamber and artificial light was provided by six 300 watt lifht bulbs susuended 6 feet above the floor. Liyhts were con— trolled in the same manner as outlined for the cold chamber and the in- J tensity of light was approximately the same for both Chambers. 1 .0 - n J , -_ 1. .».O The oririnal ranye Ol temperatures selected to be studied were ho , ° 0 o a a. . . . q . . SS , 7O , and 85 b. bince the first series Ol experiments were run at q 0 7‘ lo __‘ - .. , , 1 . q ~ 0 . w .0 w o ,5 and 05 n. and little eifierence was observed in the various pnySio- ’ W ' ' a - a - O ‘1 . 1 logical responses stuCied, it was seeided to substitute 90 r. for the o a . . . . v 70 h. group. Tne coolins capaCity of the cold Chamber was not great . . . o -1 . _, 0 ,., enough to maintain the LO r. temperature; thereiore, 50 J. was sub- stituted in its place. The exnerimental animals consisted of 2h blackface California ewe lambs and 12 native fine wool yearling ewes. The California lambs were pproximately h to 5 months of are and averaged 70 pounds when they arrived on April 15, 1957. The native yearlinfs were approximately 6 to 7 months older and averared 75 pounds. Both groups were shorn and placed on pasture where they remained until two weeks prior to the beginning of the experiment on June lb, 195?. - 19 _ At this time both groups were confined to a large pen within a masonry building with heat available where temperature was maintained at approx- imately 700 F. (plus or minus 50) with natural light conditions. This also served as a holding pen for animals between experiments which in- sured that all animals were subjected to the same environment prior to being placed on experiment and helped eliminate any pre-experimental treatment effect from altering the experimental data. Throughout the course of the experimental period, all animals were given feed, water and trace mineral salt §§:.l$2; In order to eliminate wastage and to insure accurately measured feed consumption, the entire ration was pelleted into one-fourth inch pellets. It contained the following ingredients: 1600 pounds ground alfalfa hay 390 pounds yellow corn 10 pounds dicalcium phosphate 10 pounds trace mineral salt 5 pounds Vitamin a pre-mix (5 million units) 2 pounds Aurofac 10 1 pound Vitamin 132 (625,000 units) 2018 pounds Total Each experiment included six California ewe lambs which received 1131 and were again used in succeeding experiments and one native year- ling which did not receive 1131 and was slaughtered at the end of each experiment for histological study of endocrine glands and chemical determination of thyroidal iodine. Data were collected on both the California lambs and native yearlings for weight gains, feed consump- tion, water consumption, respiration rate, and rectal body temperature. Only the California ewe lambs were used to collect data on 1131 uptake and turnover rate and l-thyroxine secretion rate. Both groups averaged approximately 80 pounds at the beginning of the experimental period on June 18, 1957 and 1&0 pounds at the termination on January 18, 1958. Following each experiment, the California ewes were given a 30 day rest in order to allow any residual 1131 to decay and to prevent any carry-over effects from one experiment to another. As a further check against carry-over effects, a stratified system of randomization was used in alloting animals to the hot and cold chambers. Analysis of variance showed that there were no carry-over effects. Since experiments were being conducted concurrently in the hot and cold chambers, a total of 12 California ewes were on experiment at all times and 12 were being rested in the holding pen for succeeding experi- ments. At the beginning of each experiment, animals were randomly assigned to individual pens within the chambers and allowed 7 days to become acclimated to the respective temperature and light condition. On the seventh day the animals were weighed and injected subcutaneously on the medial side of the right rear leg with 50 microcuries of 1131. Individual feed and water records were kept for a period of 20 days along with daily respiration rate and rectal temperature every other day. Animals were again weighed at the termination of the 20 day experimental period and the native yearling was slaughtered. External 1131 counts were taken over the area of the thyroid begin- ning 2h hours after the time of injection and continued daily until maxi- mum uptake of the injected 1131 had been reached. For output turnover rate studies, counts were taken every LB hours after maximum uptake of 1131 and continued until five such counts had been made. The counting technique as outlined by henneman.gtz 2;; (1955) was used throughout the study. All I131 counts were made by a scintil- lation counter (nuclear Chicago, model number 35-1) and a count rate meter (Nuclear Chicago, model number 1020). For l-thyroxine secretion rate studies, lSO microcuries of 1131 were injected instead of the usual 50 microcuries in order that suffi- cient count would be available following turnover rate determinations. A count was taken 3 days prior to the last count used for the turnover rate determination which was considered as zero time. Counts were taken every third day thereafter throughout the l-thyroxine injection period. Again, the technique outlined by Henneman gt; El; (1955) was used for l—thyroxine injection procedures and estimates of secretion rate. All data were collected during the period of time when the lights were on in the respective chambers and the chambers were not entered during the period of darkness. -22.. Immediately following slaughter of the native ewe which did not receive l131, the pituitary, thyroid, adrenals, and ovaries were dis- sected free of adhering tissue in a saline solution and weighed to the nearest millirram. The ovaries, adrenals and one lobe of the thyroid were fixed in 10 percent formalin, sectioned in paraffin and stained with Harris haemotoxylin-eosin. The remaining lobe of the thyroid and pituitary were immediately frozen for later iodine determination and biological assay. Chemical iodine determination of frozen thyroids were made by using the procedure of deineke gt; 2;; (l9h5) as reproduced in part in appendix S. _ 23 _ V. RJSULTS All DISCUSSION A. nffect of fimbient Temperature and Artificial Li ht on Thyroidal Uptake of I131: Counts were taken over the area of the thyroid beginning 2h hours after subcutaneous injection of SO microcuries of I13l and continued every 2h hours thereafter until each animal had reached its peak uptake of 1131. Lech count was compared with the count of a S microcurie 1131 standard prepared on the day of injection and the percent uptake of injected dosage was computed. Since a large number of researchers have interpreted a high 1131 uptake by the thyroid and the speed at which it is taken up to be highly correlited with a hiph output of l-thyroxine, this experiment was de- signed to measure the effects of ambient temperature and artificial light on both the percent and rate of uptake of 1131 by the thyroid and their relation to output rate. The results of one day percent uptakes are shown in table 1. Analysis of variance showed that significant differences (P<.Ol) existed among both temperature and light conditions. Correlation analysis revealed no significant correlation between one day percent uptake and both ambient temperature and turnover rate of I131 one-half time. The results of three day percent uptakes are shown in table 2. Analysis of variance again showed a significant difference (P<.Ol) among lisht conditions but no sirnificant difference amone temperatures; how- ever, there was a significant interaction (Pa mo.am oasm.m ma H .3oHon hampmpmamm pmpwfla ma pH .pcopcoo oSHUOH mom msaw> ewes AHoEmnpxo cm pesonm .m 0mm paw pnmfla mo nude: 0H pm mead one mo one modem ”mpoz maa.o mm am.m mm.mm aaam.m 40H m pamaa mason 0H gma.o am aH.m mo.mm qamo.m em a sauna mason NH mmH.o am a®.m mw.ow Noam.m woe N pemaa made: m mgfi.o Pm OH.m am.oH omwo.m :HH m .a 00m 04H.o 4m wa.m Jm.mm NONH.N mm H .m 0mm pmH.o om Hw.m wm.ma NOHw.H mm m .m 0mm 4ma.o mm os.m em.mm aaam.m om m .a com season pnwflos.hpon A.mEV AmEmhmv A.mnflv pflohmca mo .na hog pampcoo pgmfiox meom pflohmze deEHQd wQHUOH mcHUOH meHeOH .QH ewe mo .p: mo .pa meesaea psoopom memAmOHUH: ommwo>a Uflohhce .w: mmwhm>d erm9e>¢ .oz sOHpHpcoo seeds eaowaee mo essence mewUOH sew panama :0 named Hmaoaaapwa sew mwapmwmaeme semansa do pomp a mmdfia B. Effect of Ambient Temperature and artificial Light on Thyroid Activity as Measured by'HistOIOEical Examination: As was the case for percent iodine determinations, ewes at 8 hours of light, 550 F. and 8 hours of light, 85° F. are not included in this phase of the study since they accidentally'became pregnant and were not slaughtered. All data are based on the examination of one lobe of the thyroid gland as the other lobe was used for iodine determinations. Immediately after slaughter of each animal, the thyroid was dis- sected free of all adhering tissue, weighed, and fixed in 10 percent formalin. The tissue was embedded in paraffin, sectioned at 10 micra and stained with Harris haematoxylin-eosin. The height of one epithelial cell was measured in each of 20 ran- domly'selected follicles for each animal, using the technique of Rawson and.Starr (1938). The height of cells was measured by an eyeepiece micrometer mounted in a stage microsc0pe and calibrated with a stage micrometer. All measurements were taken under oil immersion. The results are shown in table 10. Analysis of variance revealed a significant difference (P<.Ol) among both temperature and light con- ditions. The cell height in the 500 F. group was significantly greater than.either the 850 F. or 90° F. group; however, it was not different fronlthe 55° F. group. Also the 550 F. group was significantly differ- ent from the 90° F. group but was not different from the 850 F. group. Interpreting thyroid secretion rate to be in a linear relationship 'with epithelial cell height (Rawson and Starr, 1938), the temperature effects are very closely associated with the values recorded for I131 output turnover rate and l-thyroxine secretion rate. Table 10 iffect of Ambient Temperature and Artificial Light on Lpithelial Cell height of Thyroid Gland Condition ho. 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NS.SS SS. NS. NSNS. SS.SSN SSSS. SS.NSS SH. S S S SSS NSS. SS.SS SS. SS. SSSH. SS.NN SSSS. SS.SSS SS. S S S SSS SSS. SS.SSN SS. SS. SSSS.H SS.SSS SSSS. SS.SSS SH. S S S SSS SSS. SS.SSN SS. SS. SNSS. SS.SSS SSSS. SS.SSN SS. S S S SSS *dms CH mpmm pnwflu coaSmSomm .p.onhN .S.0Iwh .p.olwx. .S K HSSSSM SSSSQ n SSSSSSMSSwSm A Ho So no Ho MO MS nSSHS Snapmummame SSSSSSSSS SS HSSSS SSS SSS SSS SSS SSS SSSSS SSSSSSS SSSSSSSSSS SSSSS HSSSSSSSSS SSS m Nfiucwmm< Sondpmnmasma Samflnsd Umdflonpmoo SS szm mo ovum nowpmnoom oGHNOHSQBWH SHSSQ copmswpmm _ 81 - Appendix I One Day Per Cent Uptake of 1131 by the Thyroid Gland of Ewes Under Controlled Ambient Temperature and Artificial Light Conditions Hours of Ambient Temperature Artificial Light 50° F 55° F 85° F 90° F Total Average 8 1.1.17 8.11 8.02 9.18 8 7.71 11.95 8.63 6.02 8 11.99 8.72 9.33 1.88 8 8.09 13.70 16.62 3.62 8 8.90 11.08 10.26 8.93 8 8.05 12.36 12.65 2.68 Total 55.91 61.92 61.51 27.78 207.13 .Average 9.32 11.62 10.25 8.63 8.63 12 8.37 5.69 7.91 8.90 12 5.88 6.26 8.26 5.81 12 7.60 7.73 9.19 7.59 12 5.36 9.86 6.67 5.32 12 8.58 7.91 7.15 6.28 12 7.37 7.68 3.38 5.56 {Total 35.12 85.13 38.56 35.86 158.27 .Average 5.85 7.52 6.83 5.91 6.83 16 5.50 13.11 7.86 5.83 16 9.58 7.69 11.62 9.38 16 8.95 8.20 9.06 8.33 16 8.92 8.61 8.09 8.92 16 5.56 10.11 7.07 8.22 Total 35.19 55.29 58.87 80.08 185.39 Average 5.87 9.22 9.15 6.67 7.73 fPotal 126.22 162.38 158.98 103.29 586.79 Average 7.01 9.02 8.61 5.711 7.59 Appendix J Three Day Per Cent Uptake of I131 by the Thyroid Gland of Ewes Under Controlled Ambient Temperature and Artificial Light Conditions Hours of AmbientTenperature Artificial Light 350° F 355° F 85° F 90° F Total Average 8 17.26 7.81 8.38 17.28 8 11.50 6.88 8.76 16.11 8 18.92 5.18 12.06 6.88 8 13.38 15.37 13.02 8.50 8 13.13 10.82 9.65 8.03 8 12.26 18.10 18.86 8.19 Total 86.85 59.32 62.33 60.91 269.01 Average 18.81 9.89 10.39 10.15 11.21 12 11.28 8.82 11.07 6.80 12 9.87 10.07 6.18 8.82 12 10.27 10.18 13.50 10.11 12 8.78 13.82 7.33 11.33 12 8.07 8.58 9.89 7.20 12 12.19 8.70 8.12 9.56 trotal 60.82 60.13 55.65 53.02 229.22 Average 10.07 10.02 9.27 8.88 9.55 16 9.79 20.33 18.98 9.02 16 16.02 9.00 15.39 18.75 16 8 .05 13.68 13.77 10.18 16 6.57 7.61 12.28 10.88 16 8.52 18.81 13.53 18.01 16 5.88 15.93 18.37 15.63 Total 58.83 80.92 88.68 78.03 298.86 Average 9.18 13 .89 18 . 78 12 . 38 12 .88 Total 201.70 200.37 206.66 187.96 796.69 Average 11.20 11.13 11.88 10.88 11.07 _ 83 _ Appendix K Maximum.Per Cent Uptake of 1131 by the Thyroid Gland of Ewes Under Controlled Ambient Temperature and Artificial Light Conditions Hours of Ambient Temperature Artificial 0 Light 350° F 355° F 85 F 90° F Total Average 8 17.26 7.81 8.13 20.68 8 12.38 11.80 10.11 19.09 8 19.36 5.18 12.06 6.88 8 13.38 15.37 16.83 9.35 8 15.08 12.19 11025 80M 8 13.63 18.28 15.58 8.20 Total 91.05 66.19 73.96 68.30 299.50 Average 15.18 11.03 12.33 11.38 12.88 12 12.88 8.82 11.56 6.89 12 10.16 10.81 6.15 8.82 12 10.27 10.23 13.50 10.28 12 8.78 13.82 8.18 11.33 12 8.07 8.71 9.83 8.78 12 18.09 8.76 8.12 10.56 {Total 63.81 60.75 57.38 55.78 237.68 .Average 10.68 10.13 9.56 9.30 9.90 16 9.79 21.91 18.96 9.95 16 19.68 10.62 16.78 15.21 16 8.05 13.68 16.89 10.72 16 7.28 9.75 16.38 10.72 16 8.52 15.01 16.93 15.03 16 6.35 17.26 22.28 15.63 Total 59.63 88.19 107.78 77.26 332.82 Anmarage 9.98 18.70 17.96 12.88 13.87 Total 218.89 215.13 239.08 201.38 870.00 Average 11092 11095 13.28 1.1019 12 .08 —88- Appendix L Zero Time Per Cent Uptake of 1131 by the Thyroid Gland of Ewes Under Controlled Ambient Temperature and Artificial Light Conditions Hours of Ambient Temperature Artificial Light 50° F 55° F 85° F 90° F Total Average 8 21.97 8.93 5.68 25.36 8 16.11 22.25 18.68 18.58 8 28.65 6.39 18.60 7.85 8 18.60 17.22 18.85 10.25 8 21.21 18.31 13.92 9.15 8 17.36 17.78 18.02 6. Total 151.90 86.78 85.71 77.25 365.60 .Average 25.32 18.86 18.29 12.88 15.23 12 18.55 12.68 18.87 6.68 12 15.25 18.01 10.80 7.95 12 10.21 12.19 12.73 9.87 12 9.83 18.00 8.68 11.19 12 9.15 9.01 12.22 11.25 12 28.76 9.78 10.78 10.60 trotal 87.75 71.63 69.28 57.50 286.16 .Average 18.63 11.98 11.55 9.58 11.92 16 12.81 21.72 22.92 11.53 16 23.06 10.18 22.32 21.58 16 9.83 20.60 16.25 11.08 16 8.13 9.11 22.63 17.81 16 10.29 23.21 18.68 17.68 16 7.85 21.17 28.28 20.82 trotal 71.57 105.99 127.00 100.82 808.98 .Average 11.93 17.67 21.17 16.78 16.87 Total 275.22 268.36 281.99 235.17 1056.78 .Average 15.29 18.69 15.67 13.07 18.68 -85- Appendix H Day of Maximum Per Cent Uptake of 1131 by the Thyroid Gland of Ewes Under Controlled Ambient Temperature and Artificial Light Conditions Hours of Ambient Temperature Artificial Light _55° F 85° F 90° F Total Average 000606000000 ‘2‘ :11 w VIC'WUJUIW IT’S" 98 .2 8.08 N Total 2 Average 5'0 K'UIC'KJJUIUI (I) NMWF’C’W U) 0 CO (1) wNNWC'E’ . h) F'UI “1:"va Total 2 Average 16 H I-' H WC WC’NNW: [00“ C’NWNWN 75 07 3013 O O O 16 16 16 nouacreacrua you» crLuLu\u\n\n ITW?’ \n\n\o$r£?\n 87 .3 3.63 Total Average H \O N N 2'13" E‘JZ’C’FC'JI" \p o 0 12‘5" rc—mwc-Jr- no N w o N o O Total 66 65 68 61 260 Average 3.67 3.61 3.78 3.39 3.61 Appendix N Twenty Da;r Body Weight Gains of Ewes Fed 32° lib. Under Controlled Ambient Temperature and Artificial Light Conditions Hours of Ambient merature Artificial Light 50° F 55° F 8 5° F 90° F Total Average 8 10 8 -1 3 8 3 5 8 2 8 3 21 8 8 8 8 3 5 2 8 6 12 8 2 8 13 7 2 8 8 8 15 3 13 Total 51 67 33 38 18 5 Average 7.28 9.57 8.71 8.86 6.61 12 8 2 8 1 12 6 9 9 S 12 8 12 12 0 12 3 6 5 2 12 8 8 12 2 12 8 8 13 2 12 2 8 5 0 Total 35 89 60 12 156 Average 5.00 7.00 8.57 1.71 5.57 16 9 8 -8 5 16 0 9 13 5 l6 6 -2 9 7 16 12 7 3 8 16 5 6 2 7 16 8 10 10 2 16 13 8 10 3 Total 55 82 83 37 175 Average 7.60 6.00 6.18 5.30 6.25 Total 139 158 136 8 3 516 Average 6.62 7.52 6.88 3.95 6.18 Appendix 0 Twenty Day Feed Consumption Per Pound of Body Height of Ewes Fed ad. lib. Under Controlled Ambient Temperature and Artificial Light Conditions Hours of Afibient Topperature Artificial Li:ht 50° F 55° F 85° F 90° F Total Average 8 .62 1.00 .53 .50 8 .76 .99 1.11 .63 8 .56 1.32 .98 1.11 8 .86 .93 .91 .88 8 .72 1.23 .88 .97 8 .53 1.01 .87 .87 8 .71 1.13 .72 .80 Total 8.76 7.61 6.00 5.76 28.13 Average .68 1.09 .86 .82 .86 12 .61 .78 .68 1.00 12 .57 .96 .96 .70 12 .75 .83 .73 .57 12 .61 .92 .81 .71 12 .88 .80 .83 .75 12 .82 .92 .95 .78 12 .50 1.02 .50 .70 Total 8.70 6.23 5.86 5.17 21.56 Average .67 .89 .78 .78 .77 16 .90 .86 .68 .86 16 .57 1.16 .88 .89 16 .71 .63 .65 .62 16 .99 1.19 .96 .86 16 .67 1.09 .79 .89 16 .82 1.19 .91 .69 16 .98 1.15 .86 .78 Total 5.68 7.27 5.69 5.59 28.19 Average .81 1.08 .81 .80 .86 ”Total 15.10 21.11 17.15 16.52 69.88 Average .72 1.02 .82 .79 .83 Note: The weight of all ewes was adjusted to 70 pounds by the use of the following formula (Brody, “Bioenergetics and Growth"): Adjusted W918ht = 70 + (deviation from 70)°73 - 88 - Appendix P Pounds of water Consumption Daily by Ewes Under Controlled Ambient Temperature and Artificial Light Conditions Hours of AmbientfiTemperature Artificial Light 50° F 355° F 85° F 90° F Total Average 8 5.18 7.20 8.29 6.18 8 5.80 8.53 9.99 . 6.82 8 3.81 13.72 10.63 12.71 8 7.70 7.70 7.72 10.29 8 3.28 9.73 9.82 10.29 8 8.72 7.96 12.00 11.78 8 8.93 11.12 8.97 8.37 Total 38.98 65.96 63.82 66.80 230.76 Average 5.00 9.82 9.06 9.89 8.28 12 3.10 7.78 6.51 13.02 12 8.93 10.18 12.71 10.23 12 8.03 9.20 10.66 8.93 12 8.89 9.92 10.58 8.99 12 6.99 9.09 8.68 10.51 12 6.10 8.06 9.86 13.33 12 8.96 11.87 8.38 9.61 Total 33.68 65.66 63.30 78.62 237.22 Average I408]. 9038 9.0).} 10.66 80,47 16 7.25 7.56 6.38 8.92 16 8.19 11.08 11.83 9.83 16 6.35 7.31 6.78 6.23 16 8.92 12.05 12.95 10.16 16 8.12 11.13 7.53 9.26 16 7.00 11.39 9.52 7.08 16 8.89 8.12 9.88 8.18 Total 86.72 68.68 68.07 59.62 239.05 Average 6.67 9.80 9.15 8.52 8.58 Total. 115.38 200.26 190.79 200.68 707.03 Average Sol-l9 905,4 9009 9055 80,42 Note: water was available 2d.31§p. for all groupd of ewes except the 500 F group which was offered water every twelve hours in order to control humidity. This may account for the differences shown by the 500 F group. Under Controlled Ambient Temperature and Artificial Light Conditions -89- Appendix Q Rectal Body Temperature of Ewes Hours of Ambient Temperature~ Artificial Light 3g50° F 55° F 85° F 90° F Total Average 8 102.9 103.7 102.9 102.0 8 102.7 103.1 105.1 102.6 8 102.7 103.7 108.1 102.9 8 102.9 103.1 108.2 103.2 8 102.5 103.6 108.2 102.9 8 102.6 103.2 108.2 103.2 8 102.8 103.9 103.9 102.7 Total 719.1 728.3 728.6 719.5 2891.5 Average 102.7 103.5 108.1 102.8 103.3 12 102.1 102.2 102.8 108.8 12 101.8 102.8 103.8 108.1 12 103.0 102.7 103.9 103.8 12 102.8 102.9 103.6 103.2 12 102.1 103.2 108.2 108.0 12 102.6 103.2 103.7 103.8 12 102.6 102.7 102.5 102.9 Total 717.0 719.7 728.5 726.2 2887.8 Average 102.8 102.8 103.5 103.7 103.1 16 102.0 103.1 102.8 103.3 16 102.1 103.3 108.2 103.8 16 102.0 102.8 103.1 102.8 16 102.5 103.2 108.7 103.3 16 102.1 103.6 103.8 103.8 16 102.8 103.1 108.0 102.6 16 102.8 103.5 103.3 102.9 Total 715.5 722.2 725.9 722.1 2885.7 Average 102.2 103.1 103.7 103.2 103.1 Total 2151.6 2166.2 2179.0 2167.8 8668.6 Average 102.5 103.1 103.8 103.2 103.1 Respiration Rate of Ewes Under Controlled Ambient Temperature and Artificial Light Conditions _ 9o - Appendix R (Flank Movements Per Minute) Hours of Ambient Temperature Artificial Light 350° F 55° F 85° F 90° F Total Average 8 82 61 135 75 8 80 90 186 89 8 31 89 158 102 8 86 118 159 158 8 36 60 187 139 8 33 99 188 182 8 82 92 109 89 Total 270 608 1039 790 2703 Average 39 85 188 113 97 12 68 89 111 181 12 59 55 111 150 12 59 83 129 188 12 82 55 150 131 12 83 50 155 135 12 56 81 138 152 12 37 89 91 123 Total 360 382 881 976 2559 Average 51 89 126 139 91 16 82 88 131 188 16 35 128 168 156 16 80 90 185 137 16 57 100 201 139 16 38 105 159 181 16 80 128 156 80 16 37 137 123 133 Total 285 732 1083 970 3070 Average 81 105 155 139 110 Total 915 1678 3003 2736 8332 Average 88 80 183 130 99 -91.. Appendix S The Determination of iodine in Thyroid Glands% - far 4- 11.68. 361’} CS: * 9. 10. A. 8. Quality KaOH and 8103 for the alkaline fusion H31 ab.) 8 5: methyl orange indicator solution Lodium bisulfite, 10$ solution, A. d. Bromine water, saturated. A. d. 1 Sodium salicylate, 53 solution Lotassium iodide, A. 8., 10; solution freshly prepared immediately before titratin; samples. 0.5% starch solution (keep refrigerated). Standard Iodate Solution) 0. N: Keirh 3.5670 grams of potassium iodate, dissolve in a small a cunt of water, transfer the solution quantitatively to a liter volumetric flask and dilute to the mark with water. then treated with an excess of RI and h280a this solu- tion liberates iodine equivalent to its volume of 0.1 h. The solu- tion is used to standardize the 0.1 8. sodium thiosulfate. lf kept in a cool place and in a glass steppered bottle the solution will retain its strength almost indefinitely. Standard Sodium Thiosulfate Solution: In approximately 0.1 8. stock solution of sodium tniosulfate, rags 09.5 H 0 is prepared by 013. solving 28.820 grams in water and diluting o the mark in a 1000 ml. volumetric flask. The addition of 10 ml. of 0.1 K. haOH per liter greatly increases its stability. *Analytical procedure for metnod publisned by a. P. ieineke, C. h. Turner, G. 0. hohler, d. D. hoover and h. B. Leezley. The determination of thyroxine in iodinated proteins having thyroidal activity. J. biol. Chem. 161:599-611, 1985. - 92 - Appendix t.“ (continued) 11. The 0.1 h. thiosulfate is standardized against potassium iodate after it reacts with an excess of potassium iodide and sulfuric acid. The procedure is as follows: To 20 cc. of standard 0.1 h. lodate solution in a 500 ml. Srlenmeyer flask add about 200 ml. of distilled water and 10 ml. of a 10; solution of potassium iodide and about 5 cc. of 1 A. sulfuric acid. The thiosulfate is added by meals of a burette. when the ionine color has faded to a pale yellow or straw color, 1 cc. of starcn solution is adced and the titration is continued until the blue color has entirely disappeared. . cubic centimeters of 0.1 l. iodate - factor for 0.1 h. thiosulfate cubic centimeters of thiosulfate - From this solution a 0.005 8. sodium thiosulfate solution is pre- pared. If the 8a0h is added the solution retzins its titre for several days. If the Naoh is omitted it must be prepared daily. Procedure Place an accurately weighed thyroid, dissected free of all adhering tissue, into a nickel crucible. To the content of the crucible add 5.0 grams of sodium hydroxide pellets and 2 ml. of distilled water. Then moisten the film of sediment remaining on the sides of the crucible by tilting. Evaporate the water by cautiously heating over a low flame. Then place the cover on the crucible and heat strongly enough to keep the melt liquid. then nearly clear add a small crystal of 8803. Continue the heating. depeat until no more efferscence is observed after adding Kh03. After cooling dissolve the melt by filling the cruicible about two- thirds full of distilled water and heating cautiously. Transfer Quanti- tatively to a 500 ml. drlenmeyer flask, using about 150 ml. of distilled water. Add 6 drOps of methyl orange solution, 1.0 ml. of 10; sodium bi- sulfite solution, and 85% H3PO8 until the color is faintly pink. Add _ 93 _ Appendix 8 (continued) Procedure (continued) enough bromine water to color the solution strongly yellow, add glass beads and boil briskly until clear. (This will take about 10 minutes.) Add 5 drOps of 55 sodium salicylate solution and cool in running water to about 200 0. Insufficient cooling causes an indistinct end point. Add 5.0 ml. of freshly prepared 10% KI solution and 5 ml. of 85% H3P0b. Titrate the iodine with 0.005 8. 8325203 solution. Use a starch solution as an indicator. (The Na28203 is freshly prepared by'diluting it in the ratio of 5.0 ml. of 0.1 8. 182125203 per 100 ml. of distilled water.) Calculation: 1 ml. of 0.005 K. ha25203 solution is equivalent to 0.106 mg. of iodine in the sample. Percent iodine = weisht of iodine 100 weight of thyroid _ 9h _ Appendix T Effect of Ambient Temperature and Artificial Lifht 0n r-H Leight and Iodine Content of Thyroid Gland Ambient Hours of height height Total Percent Temperature Artificial of of Iodine Iodine Light Bwe Thyroid Content Content (lbs.) (grams) (m§.) 50° F 8 103 2.2825 2.82 0.106 50° F 12 79 2.2071 3.80 0.158 50° F 16 88 3.2258 8.55 0.181 55° F 12 118 1.8993 3.88 0.18 55° 3 16 81 1.7220 2.22 0.129 85° F 12 93 2.1707 3.17 0.186 85° F 16 79 2.3810 87.59 2.033 90° F 8 108 2.1109 3.31 0.157 90° F 12 91 2.0885 2.72 0.133 90° F 16 183 2.0776 3.26 0.157 gppmkfix'U Affect of Ambient Temperature and Artificial Light on Bpithelial Coll height of Thyroid Gland Ambient Temperature hours of Average height of Lrtiflcial epithelial Cells Light (Hiera) 50° F 8 10.76 50° F 12 10.98 50° F 16 10.63 550 F 12 7.25 55° F 16 10.82 85° F 12 6.37 85° F ‘ 16 9.22 900 F 8 6.70 90° F 12 6.08 90° F 16 6.82 Append' V Testing Qifference Letween Two Correlation Coeffiicients Convert r to z Variance in z = l n-3 Variance of 21 - 22 = l + l nl-3 n2-3 Standard deviation of 21 - 22 = l + l nl-3 n2-3 t = 21 - 22 ttandara deviation of 21 - 22 Look up t with infinite degrees of freedom USE GE‘M i -".”’.,‘._§.' {.3 1,516.; rlUVid