EFFECT OF THE ORAL ADWNISTRATION OF CHLOROBUTANOL ON REPRODUCTION OF THE MALE AND FEMALE BOVINE Thesis for the Degru of M. S: MICHIGAN STATE COLLEGE Elmer Christian Scheidenhéim 194 7' M5195 This is to certify that the thesis entitled Effect of the Orel Administretion of Chlorobutenol on Reproduction of the Male end hemele Bovine presented by Elmer Christirn Scheidenhelm has been accepted towards fulfillment of the requirements for J; 3; “degree in _D.s'=..i.£1[_~ (3. f //¢/a1 Major/pr/ofessorfi’z DateMfiL_. _ EFFECT OF '15! CRAL ANINIS'EATION OF OHLCROBUTANOL ON BPRODUCTION OF THE MALE AND FEMALE BOVINE fay Elmer Chrietien Scheidenheln I A masxs attained to the Graduate School of lichigen State College of Agriculture end Applied Scimce in partial fulfilment of the requirements for the degee of MASTER OF SCIENCE Department of miry 1947 THESIS ACKNOWLEWENTS Ihe writer wishes to express his sincere appreciation to Dr. C. 1'. Huffman, Research Professor in Dairying for his suggestions and guidance in planning the investigation and hie criticisms and direction in preparing the namecript. In addition the writer is especially grateful to assistance given in collecting the data to 11‘. C. 1'. Clark, formerly Associate Professor of Animal Pathology and not State Veterinarian. and the many dairy breeders throught the state she cooperated in the mrnshings of animals for this study. ta. x.) . (4» --I’ f -9 1.1 ;. he TABLE 01' OORTM'I‘S Ismomcnon...................................................... RENE! OF LITmAML............................................. Reprodiction in Males........................................... Libido........................................................ nutrition factors........................................... Hornoneo.................................................... haci'.00000000000.0.0.0...00.0.0000...OIOOOOCQOOOOCOOO0.0. Factors Affecting Quality of Senen............................ Age of sires................................................ Lethal factors.............................................. Season aid cliaate.......................................... Frequency of nee............................................. Exorciso.................................................... Ration end Semen mality...................................... Vitamin C................................................... Bone factors influencing vitamin 0 synthesis................ Inter-relationship of vitamin c and other vituins.......... Vitamin 1................................................... Aline acids................................................. Minerals".............................V..................... Hormones and semen quality.................................. Characteristics and preservation of spermatozoa............... Korphologyu................................................ Air and sunlight...”....................................... pH and sperm concentration of eenen......................... storage at 'eunQOOOOOCOOOOOO0.0.0.0....OOOOOOOOOOOOOOOOOOOOO caucus-clam OOQQG 12 12 14 15 16 16 1'7 17 18 Standard of semen quality".................................. Reprodiction in Ionian...”.........................‘......... Factors Affecting Conception”............................... Libido and conception....................................... Lethal factoreandconception...u.......................... Nutrition and Concqtion...................................... Yituin 6................................................... Vitamin A................................................... Vitamin E................................................... Anino acids................................................. “13”“.000000000000.0.0.0...00.00000000000000000IOOOOO0..O. mastery of Review of Literature................................. Reproduction in the Mole...................................... Reprochction in the Tantalum”............................... OBJECT“........................................................... WERIMENTAL PROCEDJRE.... Animals Used..................................................... Feeding Procediro................................................ Bleeding Proceehre“............................................. Determination of Plasma Ascorbic Acid.. Determination of Semen Ascorbic Acid............................. Semen Collection - Spernatosoa Counts and Progressive Motility Evaluations... ”.111.an the base or LibidO..00...O...OOOOOOCOOOOOOOOOOOOOOO. an“ mar'..0...OOOOOOOOOOOOOOOOOOOOOOOOOOOOOOO0.0.0.0.....0... Mauring the 1”,"th 1n rert111w0000COOOOOOOOOOOIOOCOOOOOOO 26 28 28 28 29 32 82 REmTSQOOOOOOOOCOOOCOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOO0.0.000... Chlorobntanol ministration to Sires.......................... Libid-OCCCCCOCOOOOOCCOOOOCO00.0.0.0....OOOOOOOOOOOOOOOOOOOO... Breed» ”d Libido mprmmentOOOOOOOOOOOOOOOO0..00.0.0000... AG. and Libido IwrovwentOOOO0.0000000000000000000000.00.. Semen Qnality................................................ 'Patty. 3111.00.00...OOOOOOOOOOOOOOOOOCOOOOO00.00.000.00... ”16“” EIIOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOOCO0.00.0... Sire Sire Sire Sire Sire 311‘ e Sire Sir e Sire 'VZeL”................................................ '1601“................................................ "Shorthorn"........................................... 'lells”............................................... "IFSZO"............................................... '1hornfl................................. ..... ......... 'Ladd" 'Rem'............................................ ..... .mn"........OO..00......OIOOOOOOOOOOOOOOOOO0.0.000. Measuring the Improvement in Fertility..................... Nature of Inprovmnent in Fertility......................... Administration of Large Doses of Chlorohutanol. . . . . . . . . . . . . MOWhOlog or me %.n000000000000000000......O...0.0.0... Chlorobutanol Administration to 00's........................... smpo of 8m”...OOOOOOOOOOOOCOOOQOOOOOOOOOOOOOOQOO0.0...O... Effect of Treatment on Milk Productionnuuu................. DISCUSSION OF REwLTSOOOOOOOOOOOOOO.OOOOOOOOOOOOOOOO0.0.0.0000... Page 83 38 38 33 $38638£K3b8 3 8 5O 51 51 51 53 Page Effect of Libido in Bulls...................................... 53 Effect of Feeding Chlorotn tanol on I'ertility of Ellen”... 54 Effect of Breeding Efficiency of Oows.......................... 56 Effect of fliock Therapy........................................ 57 Effect of Milk Promotion...................................... 58 SJMMAR! AND CONCLUSIONS.......................................... 59 LITEA‘IURD CITED................................................. 61 flpmmIOOOCO0.0.0....OOOOOOOOOOOOOOOOOOOOO..0.0.0.0.000...0...... 7o Imomcn 0!! he problu of repromctive difficulties with both the ale and the femle bovine be long been one of concern to easy dairy-ea. With the advent of the proved sire and the artificial inseeinat ion program. the average age of sires need in dairy cattle" breeding is constantly increasing. In one airy herd ignorant association in lichipn in 1943, the anther found that twenty-five per cent of the sires becano in- potent. In sole cases it was he to the sire losing his libido. in other cases it was due to the low motility and fertility of the sperm. All the problens of sterility are not confined to the sales. In 1940 Butter (1:) reported that over a five year period in Iichigan seven per cent of the cows on test in dairy herd improvement association work were rnoved dne to steriliv. The nee of ascorbic acid by subcutaneous injection as a cure for bovine sterility was reported by Phillips and co-workers (5). (14), (24), (36). Additional evidence that ascorbic acid aided breeding performance in livestock vae reported by nvis and Cole (3) in their work with horses. no work of Bertree, Huff-an and mm in 1941 (15). (16) showed that the synthesis of vita-in c in dairy cave and bulls could be etimlated by orally administa'ing chlorobntonal. Since that tine Inndqnist and Phillips (31) have also reported the positive effect of increasing blood plasna ascorbic acid values by feeding chlorobntonsl. In view of the relationship of ascorbic acid notabolisa to reproductive efficiency. it seemed advisable to study the effect of chlorobntonal adninietration to sterile cows and balls. Blflfl G‘ HEW] be proud of reproduction in dairy cattle divides itself natu- rally into two fields of study. aanely: (a) that of the me. and. (b) that of the resales. Jones and co-workere (48) enunerate the reasons ftn' poor reprodiotion in sales and fueles, naeely, inadequate mtrition: poor aanageaen practices: disease conditions: inheritance of low fertility factors: and disturbances of the endocrine gland systea which in turn is affected by nutrition. heredity and disease. he literamre pertaining to libido and seaon quality in the halo and failure to conceive in the female will be reviewed. ”rotation in Ilales Reproductive failure in the nale is due either to a lack of libido or infertility of the opera. Libido he problu of ieproviag the libido of sires is rarely aentioned in the studies of sterility in the aale bovine. Nutrition factors Phillips (5) in his euidies with sterile bulls observed that their sex drive ieproved with an increase in the blood plasaa ascorbic acid values. Loptev (83) reported that peen feeds are valuable in sti-slating sex drive. he feeding of gees pass, or grass silage, or 4.7 pounds of beet fodder per 100 pounds of body weight etimlated the sex drive within about 15 days. -2... More recently Albrecht (73) reported a mrked difference in the libido of rabbits fed two kinds of lespedesa. Male rabbits fed for three weeks on lespedeza lay gown on soil fertilized with phosphorus refused to mate with does in estrun. Those fed lespedeza hay grown on soil for- tilised with both line and phosphorus continued to be somally excited by does in estrum. Males on the boy from phosphorus treated soils re- turned to normalcy when fed hay grown on soil treated with both calcium and lino. Vitamin I did not play a role in improving the libido of twenty sires fed wheat germ oil as raw: ted by Salisbury (88). He stated that the service tine was the sane as the control group which was 104 seconds per ejaculate. Tinin and Perelurian (85) reported improvement in the libido of bulls fed 800 to 1000 game of wheat germ per bull per day. Boyer and co-workers (51) reported that rats on a manganese de- ficient diet lacked sex interest. Roberts and Dawson (49) found that the feeding of arsenic solution to rabbits decreased libido considerably. Only 4 per cent ladted libido in the control group, while 89 per cent in the expaimental group lacked sex desire. Hormones Moore (27) stated that one important function of the hormones se- creted by the testis is that of regulating the behavioristic reaction of the male as evidenced by sex drive. He also aentioned that the pituitary gland is the master regulatory organ. Turner (46) stated “that low thyroid activity could cause bulls to booms sluggish in eating." He reported a marked iaprcveaent in libido of bulls fed tbyro-protein (thyroid active -3- casein) for a few days. his worker also pointed out that the effect of the thyroid feeding'nay be indirect since the thyroid gland influences the pituitary gland. he importance of the thyroid in maintaining the libido of sires has also been reported by Peterson and associates (4:3) and Spielman and co-workers (94). There was complete inhibition of libido in thyroideo- tonised sales. Tnese investigations and that of Turner definitely indi- cate that when the thyroid of the ale has become taporarily or permanently non-functional. one can expect the absence of libido. Turner stated, "that this is die to a lowered metabolic rate.“ Stone reported (18) that castrated rats which had lost all powc of libido were reJuvinatcd by albcutaneous injections of the ale sex hormone in the fore of testosterone propionate. Libido has no relationship to conception rates according to Dinhaa and co-workers (28) and (89). In 105 matings using bulls 75 were recorded as prompt. 'Ihese resulted in 0 per cent pregnancies. Ihe thirty services that were rQorted slow resulted in 50 pa cent pregnancies. Exercise Kelley (80) found that with slow bulls exorcise greatly dilinished the time of service. It is evident from this review of literature that there is a need for more informtion concerning the factors affecting libido in the male. Factors Affecting Quality of Seaen {the quality of semen produced will detanine to a large extent the breeding efficiency of the sire. Mam factors play an important role in - 4 - affecting the quantity and maliw of the semen produced. gs of sires Vhether the age of the sire has any bearing on the quality of the semen and the rapidity of service, has received sons study during recent years. Evidence that older bulls can stand heavier service has reported by Phillips (5) in which he concluded that Junior yearling bulls can easily be over-used causing them to produce low quality semen. He recommended one service every seven to ten days for a bull of this ago. With older sires Phillips suggested one service every fourth day in order to maintain a high quality of sample. Heidc and co-workere (4) using data from 8,919 services report that the service per conception for bulls of all ages was 2.67: bulls 7 to 10 years of age required 3.14 services per conception. while bulls under 5 years of ago 1.52 services per concep- tion. Bowling and associates (1) in stuaing breeding records over a period of 18 years found that bulls thirteen years of age or over were significantly loss sure breeders than bulls six years of age. heir tabu- lated data are as follows: _A_gg_ of Sire Services For Conception l - 2 years 1.50 4 - 5 years 1.56 7 - 8 years 10 - 11 years 14 - 15 years (00)!“ sit-'0 QOUI 0n theother hand Baker and associates (2) from studies with range beef bulls reported that the age of bulls did not have a significant effect on the calf crop. lo report was made on the number of services per concep- tion. It is possible that the results might have paralleled those for dairy cattle. Lethal factors It is apparent that lethal factors can reflect themselves in the breeding of females according to work done with cattle by Mead et a1. (6) who reported that two forms of sterility are sex-limited. Season and climate hiring the past four years several rather comprehensive reports have been made on the effect of season and climate on the semen of sires. Two of the reports that support the theory that the best semen is produced in the spring and the poorest in file late summer come from states in the same latitude. Phillips et a1. (82) reported. that total. sperm produced was highest inuspring and lowest in the fall. In the examination of the results of 1.135 matings they found that the higlest percentage of fertile matings occurred in April with 59.6 per cent and the lowest was in August with 40.8 per cent. Storage life of the semen in the summer time was 102.? hours. that in the winter the 126.7 hours. Otha' months showed little seasonal effect on the survival in storage. Drb and co-workers (11) as a result of a study of 879 ejaculates produced by nine bulls over a period. of one year expressed the belief that temfiramre changes are at least partially responsible for the differences in the semen quality. 'Iheir studiee confirmed those of Phillipe et el. (82) who found that semen of consistently inferior quality was produced during July. August and September, while semen of superior quality was promcsd in April. May and June. Season of the year also appeared to influence the percentage of abnormal spermatozoa. the number of abnormal spermatozoa was 25 per cent greater during July, August and Septuber than for the next highest month of the year. Similar -6- results were reported by Swanson and Herman (91). They concurred with previous workers in that the highest number of abnormal sperm was in summer. Their work is of interest in that young bulls were not affected as much by season as were older bulls. Swanson and Herman also found that the pH of semen was significantly lower in the manor than in the fall. These worka's concluded that spermatogensis in dairy bulls is not greatly affected by season. Salisbury (10). and Veetherby and associates (58) did not find a great variation in semen quality became of season of year. underson (75) reporting on semen studies in England. found that poorest semen production was in by. June and July. This is exactly contradictory to the findings on tins continent. Frequency of use fine quality of the semen of many sires has probably been lowered considerably by too frequent service. Weatherby and associates (58) found the collection of ejaculates once a day for 57 days from a two year old Guernsey bull. that the concentration of the sperm changed considerably and that the average longevity was only four days. When changed to one eJaculate once a week the volume increased from 2.6 to 4.1 cc and the longevity to 12 days. Exercise Bartlett and Perry (72) reported that bulls exercised for one hour . at two and one-fourth miles per hour resulted in increasing the volume of senen by 51 per cent. Kelley (80) found that exercising bulls at a speed of two and one-half miles per hour improved motility from a range of 5 to 45 per cut to 60 to 100 per cent. Life of the sperm was 3 to 24 hours -7- for bulls with no exercise compared to 22 to 39 hours for those with exercise. Ration and Semen Quality Vitamin 0 Phillips and associates (14) reported that bulls having a low ascorbic acid value for semen (two mg. per cent or less) were brought back to normalcy in a large majority of cases in five weeks' time by the sub- cutaneous injection of vitamin 0. Phillips also pointed out that semen ascorbic acid values above eight mg. per cent were toxic and the quality of the semen may be as poor as when ascorbic acid values are low. Just how vitamin C improves the quality of semen is not definitely known. Several theories have been advanced. Berg (32) and co-workers found that the ascorbic acid is concentrated in the seminal vesicle. has one might asmime that the spermato son are kept viable after spermatogenesis is completed by the ascorbic acid in the seminal vesicles until they are aiaculated. It is generally conceded that young sires have a higher quality of semen than older bulls. this phase of the problem was reviewed earlier by Heider (4) and Bowling (1). A possible logical explanation of thehigher quality of the semen of young bulls is given in the work of Biskind and Glick (34) who found that in bovine testicles the vitamin 0 reaches a maxim con- oentration soon after birth. D1 010 and Schteingert (45) using two to four months old white rats found a favorable effect of vitamin C on the development and function of the gonads. he injection of 50 mg. of ascorbic acid and 25 rat units of gonada- tropic hormones pa- day for a period of twenty to thirty days resulted in twice die average penis weight than when the hormones were used alone. he testis increased in size at the same rate as the penis. iiie woidits of the testis for the four different groups were as follows: Group It. in es. of testis 1. Iormal O. 361 2. Ascorbic acid alone 0.37? 3. Gonadatropic alone 0. 672 4. Ascorbic acid - gonadstropic 1.150 Sinltaneously with the report of Di Oio and co—workers. 1% and Andrews (44) reported the effect of ind acting 2250 rat units of gonadatropic hormone into a bull. here was a decrease of 42 to 67 per cent in the venous blood plasma ascorbic acid values tring the first twenty-four hour period. About two vein were required for the plasma ascorbic acid values to return to normal following the isJoction. Almquist and Andrews (42). however. in experiments where the simultaneously ianctod 50 mg. of ascorbic acid and 25 rat units of die gonadatropic hormone in 24 day old rats for four days and in 70 day old rats for 3) days produced no pester testis weight than 25 uni ts of the hormone alone. ihey concluded that thee would be no effect on rate over 50 days of ago since thq produce cospleto spermatozoa and are sexually mature at that age. Reid and Sykes (54) working with guinea pigs obtained the same results with the males. as Almquist and Andrews did with male rats. hey reported no mgmontation in testis weight when so mg. of ascorbic acid was added to the basal diet of rats that had received injections of tho gonadatropic hormone at the dosage of either 50 or 100 rat units. Vitamin 0 may indirectly fifect suen prediction by affecting the plane of nutrition. IcEenry (99) and co-workors working with guinealigs dis- -9- covered that on vitamin c deficient diets. appetite disappeared and oven- tually death occurred. they found that appetite could be maintained with as little as one-tenth of a milligram of ascorbic acid per guinea pig. Some factors influencisgfltuin c synthesis no value of sunlight and vitamin c synthesis was noted by Umxva (3?). He believed that solar radiation stimulated the organic synthesis of ascorbic acid in sun and animals. his work of King and associates (100), (101). (102) with rats showed that the feeding of several compounds stin- lsted the synthesis of ascorbic acid. The feeding of carvone. chlorobutanol a lipid fraction from alfalfa. and many other compounds with a low ascorbic acid diet increased urinary excretion of this vitamin. Bortree et a1. (15) showed that the feeding of chlorobutanol to cattle resulted in a marked in- crease in plasma ascorbic acid. Recently Linquist and Phillips (31) have shown that succinylsulfathiasole fed at the rate of 2.0 gm. per day to new born calves raised the blood plasma asca'bic acid values considerably during the first week. mccinylsulfathiasolo when fed at the rate of 15 gms. daily to cattle increased blood pla-a ascorbic acid values similar to chlorobutanol. mlorobutanol has a phenol co-efficient and could retard the develop- ment of bacteria in the rumen. Linguist and Phillips (96). however. showed that it had no injurious effect on the synthesis of thiamine. riboflavin. niacin and pantothenic acid when fed at a level of 5 gm. per thousand pounds of live- voidit for 160 days. Inter-relationship of vitamin c and other vitamins It has long boa! known that there is definite vitamin inter-relationship in various animals. 311‘. and co-workers (39) using rats concluded that one -10- avitaminosis prodicod on a diet satisfactory in every respect leads to the othistion of reserves of other vitamins. Their report on individual and multiple vitamin relationships to ascorbic acid was as follows: 1. 2. 4. Vitamin D deficiency caused a decrease of vitamin c in various tissues as heart - 35 per cent: kidney - 23 per cent: liver - 19 per cent and thyme - 27.8 per cent. Vitamin A deficiency decreased vitamin C content of heart - 20 per cent: kidney - 27.3 per cent and thyme - 24.1 per cent. Vitamin 36 deficiency - no noteworthy change in vitamin C in tissues or endocrines. Multiple depletions results in a vitamin 0 loss of - 26.6 per cent in the kidneys: 45.0 per cent in the liver: 19.2 per cent in the thyme: 12.4 per cent in the adrenals: 25.8 per cent in the pituitaries: 15.3 per cent in the thyroids. Kimble and Gordon (98) rgortod that the administration of riboflavin and vitamin A failed in some cases to improve the plasma vitamin values and the biophotometor performance. While these values were improved by file administration of ascorbic acid. according to Belasco and Murlin (33). the use of either vitamin A or O as a supplement reduced the hypermetabolism associated with hyperthyroidism. They stated that the rediced hypermetabo- lism was associated with an increased respiratory rate of the liver and the 1:1an cortical tissue. -11- Phillips and co-worlrers (38) (5) (it) showed that blood plasma ascorbic acid declines with the development of vitamin A deficiency. Twenty cubic centimeters of cod liver oil per day was required to maintain normal ascorbic values in Guernsey bulls. cotton et a1. (35) fed young bull calves on a low vitamin A ration to one year of age. The postmortem findings were as follows: degeneration of the germinal epithelium of the testis. absence of spermatozoa in the epididymie, and an accumulation of fluid in the cleft between the anterior and posterior lobe of the pituitary gland. Wolbacl: and Bessey (20) were of the Opinion that the strephy of the seminiferous tubules in vitamin A deficiency as in other epthelial organs spares the undifferentiated cells and hence recovery is possible with replacement therapm Sutton (36) work- ing with white rate found that a restriction of vitamin A caused a marked decrease in the blood vitamin C. Erb and associates (92) reported that a bull fed a low vitamin A ration for four months went blind, had staggers. and gastro-intestinal disturbances. By using testicular biopsy they found severe degeneration of the eeminiferous tubules and nearly complete die- appearamce of germ from the lumena of the tubules. reading of 60.000 to 150,000 international units of vitamin A per (my resulted in recovery within two to five months. Just before semen prodiction ended. the number of ab- normal sperm was four times that of a healthy bull. Guilbert (95) in his review of studies of nutritional reproductive failures also mentioned testicular degeneration due to vitamin A deficiency. Phillips (68) reported that cellular sloughing is noted in vitamin A deficiency but spermatogenesis contimes. The condition could also be re- paired after 60 to 90 days of vitamin A administation. More recently Moore and Cotter (97) in a well controlled experiment -12- with young bulls seven to four teen months of age and with older sires studied the effect of vitamin A intake on plasma ascorbic acid values. They concluded that only in extreme vitamin A deficiencies would there be a decrease in the amount of vitamin C excreted per day era depression in the blood plasm values. They predicted that extremely high levels of vitamin A intake migt stimlate vitamin 0 synthesis. They further stated that it remains to be shown that low carotene intakes of animle fed normal winter farm rations affects vitamin 0 synthesis or results in breeding defici enci es of sires. Vitamin I That vitamin E has no value in improving semen quality has been re- ported by Oullickson and co-workere (87). ‘lhey fed seven ales on a ration devoid of vitamin E and found that growth, physical and sexual development were normal in every respect. In the work of Salisbury (88) sires fed vitamin E in addition to the normal ration failed to show an increase in the volume of semen, sperm concentration or motility of the sperm. Timin and Perslurian (85) on the contrary reported decidedly favorable results from the feeding of vitamin E, in the form of wheat germ concentrate. 'iheee workers in comparing five bulls fed the concentrate to five bulls fed the basal ration reported a 31.5 per cent increase in spermatosoa volume. 63.0 per cent increase in sperm viability and a 32 per cent increase in sperm concentration in favor of the wheat germ. The beneficial effects were noted 25 to 30 days after feeding started. They recommend 800 to 1000 gms. per bull per day. Amino acids Whether dietary protein has been a factor in producing high quality -13- semen has been under investigation for some time. In stucb'ing sterility problems Gunningham and Hapkirk (22) found that feeds which were too my in protein content brought on sterility. Using rats. the feeding of pro- tein diets of 65 to 82 per cent and those containing 15 to 18 per cent protein from maize and gelatin caused sterility, which was associated with a degeneration of the testis. These results were believed to be due to an amino acid deficiency. Webster (23) reported a high evidence of sterility and abnormal spermatozoa when the crude protein of the grass was as big: as 35 per cent on a dry matter basis. Holt and coworkers (50) working with humus found that a diet deficient in arginine over a ten day period cased a reduction in the number of spermatozoa to one-tenth of the normal values. Apparently strephy of the spermatogenic tissue took place. A definite standard for a ration for breeding bulls has been worked out by Pankevic (74). He stated that a sire should get 411 gms. of digestible protein for each 500 kgs. of body weight: furthermore the feeding of the animal at the rate of 162 gms. of protein per mating should keep the bull in good breeding condition. The use of pro teins of animal origin in the ration of sires was made by Laptev (83) and hernov-Ugrjunov and Laptev (21). The use of blood meal and skimmed milk weed a makred improvement in the sperm concentration. These feeds apparently influenced spermatogenesis since a noticeable effect took place in 1'7 to 37 days. Concentrates of plant origin rated next to the two pro- teins of animal origin. They mentioned that green grass alone caused a de- crease in the amount of the ejaculate. mirymen in many regions have turned to alfalfa as the sole source of hay. This legume is low in sulphur containing amino acids. Jones and associates (48) using two groups of bull calves starting at 5 to 7 months of -14- age and continuing to 32 to 41 months of go compared a ration of alfalfa hay. salt and disodium phosphate. to one of alfalfa hay. salt. disodium phosphate and a grain mixture of skimmilk powder and eat groats. In com- paring the initial motility of the semen of the two groups it was found that tho alfalfa hay group had only 52 per cent of the semen score ex- cellent. compared to 72 per cent for the supplemented hay group. Both groups of bulls. however. were fertile. iho bulls fed hey alone required only 1.37 services pa conception compared to 1.54 for bulls of the supple- mented group. ihe Jersey hills prodiced 3.75 so of sun per eJaoulate (sowed to 6.89 for the Holstein bulls. oniihert (95) stated that when feed restriction takes piece which retards growth in the young or weidtt less in adults. testimlsr atrophy may occur in the male. mnerals Lardy and associates (52) fed a group of bull calves to 18 months of age on a grain ration containing only 28 parts per million of manganese. he semen promced was not as high in quality as flat of the ration high in this element. They mentioned. however. that most farm rations are higher ih manganese that the one they fed. Johnson (47) be shown that with pigs "promotion was not mioeessfnl with one-half part per million of manganese. hpplementing the ration to bring the mnganeso content to six parts per million brandit back normal reproduction. Mr and associates (51) in investigating the effect of manganese deficimit diets in the rat anclnded that it did not reult in a lowered ascorbic acid content of the various tissues. However, the rats on the deficient diets showed complete lad: of spermatids and spermatosoa. he -15.. nuclear division was decreased or absent resulting in a decrease in the aims of the individual ubulos. i'he lumen of die tubules was filled with degenerating cells instead of spermatozoa. these workers also noted that ascorbic acid feeding had no powth stimulating powers when added to low manganese diets. i'hat iron is present in spersmtosoa was reported by little and sun (29). they reported that the non-hemin iron of tun spermatozoa was estimated at sixty per cent of the total iron. In an extensive study using three thousand rabbi ts Roberts and hwson (49) found that arsenic fed in the form of lbwlers solution had in- Jurious effects on the reproductive system. The treated males prodiced less suen and had fewer sperm: however. there was no difference in the motility of the sperm. ihere were also fewer young in the litters sired by the treated males. the sulphur contut of sperm is quite hid: and varies com 1.6 to 1.8 per cent ”cording to little and O'dell (77). Hormones and semen quality Haw investigators have mentioned fie fact that the pituitary gland plays an important role in reproduction. Phillips (14) stated that this gland plays a role in normal repromction in he bovine. kites and co- workers (25) and Phillips (68) found fiat spermatogenosie dependent upon normal functioning of the anterior pituitary. In fact the removal of the gland cased mar-atogenesis to cease. mat the thyroid has no effect on the quality of the sperm has been effectively duonetrated by Peterson and others (43) and Spielman and associates (94). In both reports it was stated that normal reproduction took place from the semen of thyroidectomized males at four months of age. The gonads developed normally and there was no deleterious effect on the volume of semen. longevity and fertilizing ability of the sperms. It was also found that the blood plasma ascorbic acid values were normal. hill (64) found that in hyperthyroidism there is an increased re- quirement fa' vitamin 0. Characteristics and preservation of spermatozoa Mogpholog he relation of the morphology of the sperm to fertility has been suidied by Swanson and Herman (12). They found that coiled tails and pyriform heads did not reduce fertility up to :1) pa‘ cent of the total volume of sperms. Salisbury and associates (60) (81) showed that the methods of cooling influences the morphology of the sperm. 'Iheee workers found that in order to avoid inJuring the vitality of the spam that it was necessary to cool semen by reducing the tanperature at a rate of 6° C. way 10 minutes to a storage temperature of 5° C. may also report that the rate of warming to bring back motility should be rapid. In fact. thirty degrees per minute was not harmful. lur- thermore. Salisbury and Mercier (90) found that to determine accurately the amber of morphologically abnormal spam that only 100 needed to be counted on a slide. my and associates (70) reported that a large number of abnormal sperms in semen is an indication of low quality as Judged by oxygen con- sumption. Beck et a1. (65) devised a method to quickly evaluate semen low in ongon consumption. Studies by mugierty and Ewalt (78) found that there was very little correlation between the abnormal forms and the fertilizing ability of bulls. Thq found that 'croohod tails were the principle vpe of ab- normality. Laeiq and Bogart (71) in making a careful etudy of the location of the bending of the tail found the bonding was at the posi- tion of the protoplasmic droplet on the posterior region of the midpiece. Air and sunligt Both llaoLeod (79) and Letard (o?) pointed to the fact that eenen cannot be exposed to air and sunlight very long without killing the sperm. dew and associates (9) working with rm in high altitudes concluded that the altitude does not affect the spermatozoa tut probably has some detrimental effect on the reproductive system. pi and sperm concentration of semen misbury (59) commented on the fact that the hydrogen ion concen- trate is of some value for estimating the quality of seen. He stated that the pH of the semen ranges mm 5.0 to 7.2 with a mean of 6.5. hvis ‘ and 'illiams (56) reported pH values for eJaculates taken in accession. be first. second and third ejaculates had pH values of 6. B5. 7.23. and 7.21. respectively. Salisbury (59) stated that the number of spor- usually varies from 330.000 to 1.700.000 per cubic m.m. and that good quality semen should have 500,000 or more sperms per cubic m.l. hvis and 'illiams (56) reported concentrations of 826.000 per cubic m.m. for the first ejaculate. 635,000 for the second and 337,000 for the third. Salishury and hvis and lillisms need he hemo-cytometer for sperm counts. Schaffner and Andrews (52) de- veloped a method of counting spen using hematocrit hibes since in using the hue-cytometer one out of nine samples varied. in count as much as twenty per cent. hey obtained more uniform results with their method and gave an average count of bovine semen as 960,000 sperms per cubic mm. Storage of semen Suen can be stored either with diluter or undiluted. Undeer erg and nvis (57) found that the motility of diluted and undiluted semen was saintained for 200 hours. lurthermore they reported that the egg yolk salts of diluters had little effect on the fecundity of stored samples. fleet investigators are in agreement on to temperature to use for storing semen. nvis and co-workers (66) found that 35° r. gave a better storage record than temperatures of 40° or above. Salisbury (69) found in his work that the best storage tesperatnro was 5° C. which is close to the temerature recommended by mvis and his associates. Standard of semen quality Several workers have attaptod to establish standards to use in estimating the quality of semen which is satisfactory for normal conception values. 8women and Herman (93) using 23 sires reported that semen to be of good quality diould have a progessive motility of d5_per cut or more. Lasley (69) corrleated the motility and fertility of bull sperm. In salples with less than 40 per cent motility the fertility was 54 per cent cospored to 69.4 per cent when the motility was 61 to 70 per cent. In com- paring the quality of canon of the first and second ejaculates. Davis and lilliams (56) found that the first ejaculate required 1.33 services per eon- cqtion and the second ejaculate 1.25 services. Another standard of motility for age can be taken from the work of 'hvis and co-workers (66). ihoir roail ts with storage of undiluted send: at 35° were: -19- Stage 52 loLof Samples llotilig fresh 4? 81 per cent 24 - 28 hours 12 60 per cent 48 - 52 hours 33 53 per cent 72 - 78 hours 10 47 per cent Loproduotion in Females Soveral workcs have published data isich give the livestock breeds a standard with which to moanre reproduction in the female bovine. Raider and coworkers (4) reported that with dairy femles of all breeds there was little apparent effect of age upon breeding efficiency up to nine years of ago. hey also found that breeding cows for a 365 day record in- stead ef a $5 m record did not decrease the breeding efficiency. Using 432 cows in this study. they reported that the cows bred to calve at 15 months interval required 2.54 services per conception compared to 2.69 services per conception for those bred for 12 months calving interval. they also reconendod that in commercial herds cows do not warrant more than five services. Casparing heifers bred for the first time and older cows. they found that the heifers required 3.23 services pa- conception com- pared to 2. 76 for the older cows. Similar data showing that virgin heifers do not conceive as readily as older cows was reported by Bowling and collaborators (l ). hey rqoorted that to number of services required for conception were as follows: first 2.79: second 1.86: third 1.79: fourth 1.82: fifth LE). and for all services airing life time 2.02. ihq used 708 funles in this study. Work- ing with beef cattle Rhoad (8) concluded that there was not much difference in the rate of conception for heifers and cows. - go .. Factors Affecting Conception Libido and conception Unless cows have regular estrus periods conception cannot take place. A report by Pinetoeu (84) stated "that the feeding of ergot of rye had aphrodisiacal powers.“ By feeding thirty grams to cows and sows after fasting the animals came in heat in a few days. If they were not mocsssfhl the first time the dose was rqieated. accessful remilts were obtained from sixty to one lntndred per cent of the time with cows. A. lack of sex interest was produced in female rats by feeding a ration low in manganese according to Boyer and co-workors (51). Lethal factors and conception lead et a1. (6) using six bulls tested for deleterious recessive genes which were associated with female sterility. two different forms of fouls sterility that prevented heifers from conceiving or producing offspring were found. All the affected animals in the Jersq herd mani- fested normal heat periods. the sterility found in the Holstein breed was associated wifii abnornl estrous cycle. usually complete absence of heat. htri tic n and Concepti on Vi tamin 0 According to Riddoll and vhitash (so) and wants (41) cows synthe- sise mifficient vitamin c to supply their needs. Phillips and'co-workers (24) reported that occasionally cows do not synthesise mifficient quanti ti es of vitamin 0 and consequently diow a rediced conception rate. they found that "hard to settle'' cows did not show an elevation of ascorbic acid values during estrum compared to normal cows which showed a 75 per cent increase -21- he treabat of sterility by subcutaneous inJections of ascorbic acid resulted in conception in so per cent of the cows. That vitamin C is concentrated in the corpus luteua was reported in studies of humans by Ley (so). He treated 10 wonen who had experienced two or more abortions by the following procedire: early in the aibsequent pregnancy each women was given injections of vitamin 0 until the blood was saturated: than smaller doses were given to the end of the sixth or seventh month of pregnancy. All tn woaen delivered living children. Biekind and oiiek‘ (55) working with cows were of the opinion um the corpus luteum is physiologically related to the anterior lobe of the hypophysis and that it contains vitamin c in approximately the same con- cutraticn. hey made determinations of the vitamin C ccntnt of the corpus luteua of cows at various stages of the estrus cycle. hey found that when the organ was fully developed that there was 1.4 lg. of ascorbic acid per gn. of tissue aid following regression it contained only 0.8 lg. per gm. hiring gestation the vi taain 0 level remained at 1.5 to 2.2 ag. per ga. of than for seven months and decreased to 1.1 gm. in the eigith south. ihe authors flicught that there Ildlt be a relationship between vitamin c and progesterone. Studies have been conducted by Erb and associates (44) on the effect of the gonadatropic hormone on the blood plasma vitamin C of cows. the in- Jection of 2250 rat units of the hormone caused a an to 50 per cent decrease in the ascorbic acid values during a 24 hour period. Almquist and associatee (42) working with rats failed to find any effect of ascorbic acid administration an the gonadatropic hormone. more was no increase in the size of the ovaries or the uterus. Sutton and co- workers (17) reported hat the estrogenic hormone caused an increase of -23... vitamin C in both the blood and the urine. Working with fenle guinea pigs Reid and Sykes (54) were able to potentiate the inJections of gonadatropic hormones by adding 50 mg. of ascorbic acid to the basal diet. ihe weight of the ovary was 1.38 times that obtained when gonadatrepic hormone was added alone. Vitamin A The effect of the daily intake of vitamin A in relation to con- caption rates was studied by Kuhlman and Gallup (53). they found that the minim daily carotene requirement of dairy cows for normal reproduc- tion appeared to be 40 to 45 micrograms per pound of body weight. 'nie conception rates of cows fed different levels of carotene intake were as follows: Days red Number of Carotene Services Before Service Cases Intake Per Conception 90 21 33-39 micrograms 1.99 90 23 40-59 " 1.35 90 15 100-353 " 1.21 Moore and Cotter (97) feeding dairy heifers on a carotene free pain mixture. 10 pounds of wheat straw and carotene in alfalfa meal equivalent to 42 micrograms per kilo of body weight. found-no particular relationship between the stage of gestation and the excretion of ascorbic acid. rive grams of chlorobutanol fed to two of the heifers resulted in four to eigit fold increased ascorbic acid excretion in the urine. The ascorbic acid values of the blood of pregnant heifers fed a low carotene ration did not increase, however. thq excreted large quantities of it. Cuilbert (95) stated “that vitamin i deficiency results in death of the fetus or the birth of nonviable young." -23.. Bertree (51) reported that a fair year old heifer receiving only 14 micrograms of carotene per pound of body weigit for a considerable length of time had vitmin C values within the normal range. He observed that animals on a low vitamin a ration did not have a wide variation in vitamin C values. Vitamin l he effect of vitamin B on conception was studied by Henke (86). min- v-eight cows with irregular breeding behavior were fed sprouted oats. Of these 82 per cent produced calves. Thirty-seven cows with similar irregular breeding record had a 76 per cent calf crop. The cows received you feed in addition to the sprouted cats. The author concluded that the feeding of sprouted oats was of little value as a treatment for sterility in cows. Gnllickson and co-workers (87) in a study using 15 cows fed a ration totally devoid of vitamin I reported that the sexual development was normal in every case and only two of the heifers required more than cue service. Amino acids no value of proteins for accessml reproductions has been definitely indicated by (hilbert (95) in which he discovered that protein deficiency causes a cessation of ovulation in the female. Ginningham and Hopkirk (91) reported that excessive protein of 55 to 82 per cent in the ration cased some sterility in female rats but not to as great an extent as with males. -24- linerals According to Cuilbert (95) phosphorus is essential for repro- thction. minlen and Roux (7) also reported that in areas where the soil was low in phosphorus that the fertility of the cattle was low. mese authors were of the opinion that forced his: milk derands are reflected in the gnital tract resulting in the failure to show the physiological symptoms of estrum. lauganese deficiency and its relation to female conception rates have been rqaorted by Lardy and associates (52). Johnson (4?). and Boyer et a1. (51). the latter workers using rats reported that the females on low manganese diets showed a delay of twice as long in the opening of the vaginal orifice as did the normal animals. be ill effects of Fowlers solution iiich contains arsenic upon rabbits was discovered by Roberts and hwson (49). the services re- mired for conception increased from 1.08 to 1.78. The litter size was also reduced and thus was a such hiQier mortality of the ytnng. i'he destructive effect of potassium iodide upon the vitamin C of the blood has been reported by Brown and associates (53). hey stated. "that it might be possible for this mineral to rediee the breeding efficiency of a bed.“ aimmary of Review of Literature neprcdiction in the hale he factor affecting libido in bulls has not received very mch study. It appears that a ration tdiich improves vitamin C snythesie or the indection of ascorbic acid will aid in maintaining sex drive. ‘me data showing the effect of feeding green feeds and sources of vitamin B on libido in the male is contradictory. he feeding of manganese deficient diets to rats resulted in sterility in the males. lite feed- ing of arsenic «proceed libido in rabbits. A marked lack of thyroid activity is associated with impaired sex interest. Exercise may increase libido in the bull. fine quality of semen qipears to be lower bring the summer months. Exercise and the frequency of service uy influence semen quality. ihe age of the sire does not appear to have any great effect on semen quality. until the bull has reached seven years of age. after this age semen quality tends to be lower. nations appear to have some influence on sea quality. rectors iiich improve ascorbic acid metabolism may improve semen quality. ihis vitamin does not migment the effect of gonadotrophic hormones. lurked vitamin A deficiency results in an increase in the amber of services per conception. The experimental work conducted in the United States indicate that the feeding of wheat germ oil as a source of vitamin E is of no value in maintaining semen quality. Recent work indicates that the feeding of animal proteins as supplement to bull rations increases semen quality. Kanganese is the only mineral element that has been studied to any extent in relation to ease quality. Sterility was prohced on a very low manganese ration. Farm feeds contain more manganese than were used in these experiments. lite feeding of chlorobutanol and succiwl sulfathiasole to bulls increased plasn ascorbic acid values and consequently my be of value in the treat-at of sterility. Hormones have been used in an attempt to increase semen quality. be evidence indicates that the anterior pituitary gland is necessary for spermatogenesis. he throid gland has no effect in this respect. Several investigators have studied the preservation of semen. lost workers are of the opinion that a storage temperature of 5° C. was optimin. Values for good quality of semen varied someshat. but it appears that the initial motility miould be 45 per cent or more. Reproduction in the female Two studies have been made on the factors affecting libido in the female: one with the bovine and one wimthe rat. he use of manganese and the ergot of rye appeared to have some value in increasing libido in the female. Studies on the factors influencing conception in to fuale have not been as extensive as those with the fertility of sires. ihis is die to the fact that reprofictive problems of the female bovine affect only one individiel. whereas infertility in the sire affects the entire herd. female bovines do not lose their reproductive efficiency as early in life as the males. lo diffa'encs is noticeable up to nine years of £30. Heifers -27- require nore services for the first conceptiai than ucceeding ones. Lethal factors which decrease fertility m be inherited. Vita-in o appears to be cloeely related to conception. The vitamin C onntmt of the corpus luteun is hid: early in gestation period. he ad- ainistration of vitamin 0 along with gonadotropio hornones in giinea pigs resulted in an increase in the sire of the ovaries. Vitanin A does not appear to be a contributing factor in reproductive failure in the fenle bovine under farn conditions. Vita-in 1 ie not a factor in bovine fertility. hosphorous and manganese deficiencies any cause breeding difficulties. 0n the other hand the feeding of exceseive counts of arsenic and potassina iodide caused reduced fertility. - 23 - OBJECT ihe obJectives of this experiment were to determine: (a) the effect of increasing the blood plasma ascorbic acid by the use of chlorobutanol or some other stimlating effects of the drug to improve the libido of the mle bovine: (b) to neaeire the improvement of the quality of the spermatozoa of male bovine with poor breeding histories: and (c) to stuw the influence of the drug on the conception rates of the |'hai'd to settle" female bovine. MIME PROCEIIJRE Animals Used lhe bulls and cows used in this experiment had histories of having breeding difficulties. therefore. they could not all be housed at the College Experimental barns because of lack of room and labor to take care of that. “any of the animals were left on the farms of the owners and data were acquired with their cooperation. A few of the animals in the experinent were iron the main breed- ing herds of the M17 Departments of Michigan state College and Rutgers Univer si ty. feeding Procedire he anilnls were fed the chlorobutanol in the grain ration. me entire quantity for a one-day dosage was fed in the grain at one tine. he basic dosages were fed at the rates indicated by Bortree et el. (16). Variations from these amounts were made for various animals largely on a weight basis. The chlorobutancl was supplied to the cooperators by the - 29 - Michigan Agricultural Experiment Station. Instructions for keeping the records were also furnished. Bleeding;Procedure The animals were bled from time to time during the experiment when it was convenient to bring the blood samples to the laboratory within two or three hours time. Because of war time restrictions of travel. it was impossible to bleed many of the animals as often as desirable. In all cases the smples were drawn in 25 ml. tubes. ‘me antico- agulant used was five or six drape of a 20 per cent solution of potassium oxalate. 'Ihe blood was mixed with the anticoagulant by turning the stoppored tubes and over and in a slow manner. 'i'ney were then placed on ice and protected from the light while being transported to the laboratory. Determination of Plasma Ascorbic Acid The procedure used for the analysis was that described by Mindlin and Bitler (19) with certain slight modifications for the micromethod. The usual practice of adding a five per cent solution of potassium cyanide to the blood was omi tted as several workers had reoor ted that it gave no added protection to the ascorbic acid. The samples were centrifuged as soon as they reached the laboratory. 'lhe filtrate was prepared by pipetting two ml. of the plasma into a test tube containing two ml. of the distilled water. To this was added four ml. of a five per cent solution of meta- phosphoric acid and mixed by shaking gently. The mixture was filtered through a number two matman filter paper and the filtrate collected. The next step was altered somewhat from the usual micromethod as the only instrument available was for the micromethod. Only one ml. of the filtrate -30.. was added to one ml. of the dye and the amount of dye reduced was measured colorimetrically. To remove the error of a visually undectable turbidity of the filtrates the samples were reduced further after making the reading for the sample. Corrections were then made for the final reading. In calculating the results the original formula was used and the results multiplied by four to compensate for using only one ml. of filtrate. The indephenol-acetate solution was made up as described by Mindlin and Butler (19) except it was diluted so that the blank analysis gave a galvanometer reading of 70. This weaker dye solution gave a. wider range between the blank reading and the sample reading and reduced the error in the results. Diplicate readings were made in a majority of the determinations on each filtrate and an average of the two used in each calculation. The I value which was used in the calculations was checked at various intervals by the use of standard solutions made up in 2.5 per cent metaphosphoric acid solution with crystalline ascorbic acid. Determination of Semen Ascorbic Acid One ml. of semen was diluted with nine ml. of distilled water. Two ml. of the diluted semen was used. and the determinations for ascorbic acid proceeded from this point Just the same as for blood plasma. The final value was always 10 times greater in mg. per cent than for blood plasma. Semen Collection - fiermatozoa Counts and Progressive Motility Evaluations All samples of semen were collected by using an artificial vagina. The semen was collected in a pyrex centrifuge tube. Immediately after -31- collection flue semen was placed in a receptacle containing water at a temperature from 95 to 100° 1'. Upon arrivirg at the laboratory one ml. was taken fro ascorbic acid determination and another small quantity was used for making smears and for motility studies. The remainder of the semen was kept in a tightly stoppered centri- fuge bibs. To avoid temperature shock «he to too rapid cooling the following procedure was need. he tube was wrapped with paper toweling as insulation and then placed in a large pyrex test tube. This tube was placed in a beast of water at 55 to 60° F. be beaker was then set in a refrigerator at 40° I. Hi thin the insulated cnetrifuge tube fine semen cooled slowly at the rate of qaproximately 5° C. every 10 mimtes until the storage temperature of 40° r. was reached. it 24 hour intervals for three nocessive days small samples of sasen were removed from the original sample by pipette and studied for motility. After removing the sample and placing it in a small test tube, the cold diluent which was at the sme temperature as the semen was added. 'men be diluted semen was warned rapidly to 95° r. and motility values made imediately. Spermatosoa counts were ads at various intervals by using the hemocytometer and following the red blood oountprocednre. Early in its study the motility estimates were made without dilution. 1 drOp of saon was placed on a slide. covered with a cover slide and values established at 450 x magnification. A majoriw of the determinations were done by diluting with a nutrient diluent described by Willett et a1. (81). IAiring the early part of the experiment motility values were estab- lished in four groups as -. --, ---, and ---. The first group repre- ssnted values from so to 0 per cent. the second AD to 60 per cent. the -32- third 60 to 75 per cent and the fourth over 75 per cent. Only original motility was recorded at that time. Later in the experiment all values that were above five per cent were recorded to the nearest five per cent. lhis figure was arrived at by evaluating motility at five different areas on the slide and averaging them and placing the value at the closest five per Cents Evaluating the Mes of Libido lhe procedure used was to note the length of tin in minutes or fractions of a minute that elapsed from the time the male was stood at a distance of two or three feet behind the female and the completion of the service to the cow. Semen Smears Occasional smears was made of the fresh semen to be used for morphological studies. A small drOp of semen was placed near the end of a glass slide. A second slide was used to draw the draplst evenly across the glass by contacting the drop with the end of the second glass. After the semen was dried it was set with alcohol and then stained by using a three per cent rose bengal solution. gearing the Improvement in Fertility Breeding performance of a sire was measured by the number of services per conception. To accurately refl set this basis no cow was considered pregnant until she was pronounced safely with calf by a veterinarian or had actually delivered a calf. - 33 .. RESULTS Chlorobutnnol Administration to Sires Libido hiring a three year period fifteen slow breeding sires were studied to determine the effect of the oral administration of chlorobutanol upon libido. The detailed results with each sire will be found in the Appendix. The rate of feeding had to be kept at the five gram level for most sires. Only one large Brown Swiss bull could be kept at the ten gram feeding level without manifesting muscle incoordination due to the anaes- thetic effect of the drug. It appeared that many of the sires were affected by the drug althmgh they did not lose their muscle coordination. This effect was one of drowsiness. Most owners reported that their libido was much better a week after the feeding of chlorobutanol was discontinued than while the feeding was being done. Only one sire had any ill effects from eating the drug. In this one case the individual developed a foaming at the mouth when consuming only five grams of the drug pa' day with the ration. Recovery of libido was attained in 76.6 per cent of the cases. “his time of service was reduced from an average of forty-five to three and seven tenths minutes per bull. The results also indicate that a feeding period of 4) days must be anticipated with a total consimption of one-half pound of the drug. Bread and Libido Improvement Whether sise was a factor in the speed of recovery appeared as an -34- important part of this study. It was discovered that most bulls could not be fed consistently more than five to seven grams of chlorobutanol without losing their mscle tone. lherefore. it may be assumed that a heavy Holstein bull m require more days of feeding to get the same re- sults. Table II presents the data according to breed. The data for the Guernsey bulls cannot be considered representative since there were only two animals in this group. Ihrthermore, one of the sires of this breed was 14 years old and was fed in excess of the normal period of time in attempting to get a response. A comparison of the Jersey group and the Holstein group shows that the Holsteins were twice as large in weight as the Jerseys and that it took an average of 14.7 more days with 41.1 per cent more chlorobutanol to get the same results. It was interesting to note that there was very little difference in the age of the two groups and that they both had an average return to normal libido of 75 per cent. The Holstein group made more recovery when one consideres the time required for service: however, they were as slow originally as the Jerseys. go and Libido Improvement To determine if the factor of difference in age would have an effect on the degree of recovery the data were assembled in two groups. Eigit sires two years old or younger were placed in one group and seven sires three years old or over was placed in another. he data are pre- sented in Table III. 'ihe older group of sires with an average age of 6.6 years were twice as heavy as the younger group and consumed dosages on a daily basis one-third larger tlan the younger group. he older group took only two -35- more days to recover than the younger group. The main difference between the two groups was the percentage that returned to normal. Only 56.2 per cent of the older group recovered compared to 94 per cent of the younger group. 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Seven of these sires were kept on the owner‘s farms, since the bulls were needed for service. ihe data were semred from 10 separate experimental periods. Two sires were studied for resmnse to chlorobutanol. in two separate trials. All of the sires used had low conception rates before starting on the experiment. Conception rates were determined on the semen of a majority of the sires used in this study. in order to meamre the effect of chlorobutanol feeding on sperm fertility. "Patty" bull: 'Ihis 900 pound two-year-old Jersey sire was fed chlorobutanol as a treatment for sterility. me brseding history airing the seven months prior to the feeding of this dmg was 5.33 services per conception (32 services and 6 conceptions). Diring 4-3 days following the beginning of the treatment, only 1.50 services were required per conception (6 services and 4 conceptions). Tne effect on blood plasma ascorbic acid and sperm motility are shown in Table VII of the Appendix. The plasm ascorbic acid was 0.22 mg. per 100 ml. on February 28. and gradually increased following the admnistration of 5 gm. of chlorobutanol daily until a peak value of 0.51 mg. per 100 ml. was observed on March 22. The quality of the semen also improved as was indicated by increased sperm motility. Chlorobutanol was discontinued on April 12. [there was a marked Map in plasma ascorbic acid but the sperm motility increased to '75 per cent where it remained for the next five months. ibis sire was purchased by the Michigan Agricultural heriment sta» tion in November, 1941 and placed on a long time experiment. In Mauch.1942 -40.. the ascorbic acid plasma values had drapped to 0.10 mg. per cent. his low value indicated that this sire was subject to ascorbic acid deficiency. It also appeared that the original motility may not be the best meamre of the fertility of the sperm. '1!) study this problem further it was decided to make motility determinations at intervals of 24-hour periods for three days. he ascorbic acid content of the blood plasma and semen and the pH. volume and motility of the semen for one year are tabulated in Table XVIII of the Appendix. hiring the second chlorobutanol feeding period. '75 days were required to bring the sanen of this sire back to normal. fibers were no experimental cows available for service at this time. The sperm count returned to a normal level in a period of three weeks time after the beginning of chloro- butanol feeding. Almost ten weeks. however, were required to obtain nornal liveability of the sperm. A study was also made of the morpholog of the sperm in order to determine the possibility of other casuative factors contributing to poor fertility in this bull. The results are shown in Table II in the Appendix. At the beginning of the second trial there were 76 per cent abnormal sperms and a majority of these had coiled tails. After a period of four and one- half months of chlorobutanol treatment the abnormal sperms had decreased to 21 per cent. 'Ihese abnormal spam had coiled tails. Two and one-half months after feeding of chlorobutanol had been discontinued the per cent of abnormal sperms had again increased to 43 per cent. . 811‘s ”1645": Ihis sire was a l500-pound three-year old Holstein-biesian bull which had a poor breeding history. the number of services per conception for the ca. 11 months prior to chlorobutanol treatment was 3. 8. Data showing the effect of chlorobutanol on plasma and semen ascorbic acid and germ motility are shown in Table I of the Appendix. 'Ihis sire started with a blood plasma ascorbic acid value of 0.28 mg. 135‘ cent which is considered normal. The motility of the semen, however. was very low. A mximm ascorbic acid level in the blood was reached within three weeks time. In three months time the motility of the semen had re- turned to a normal value. Five months after the feeding of chlorobutanol had been discontinued the semen ascorbic acid values were declining and the number of services per conception had increased to 11. reading of chloro- butanol was again started. In two months time both the ascorbic acid values and motility had returned to normal. In Table II of the Appendix the breeding efficiency of this sire is evaluated. During the first five month feeding period. 13 services resulted in five conceptions or 2.60 services per conception, compared to 42 services and 11 conceptions or 8.80 services per conception prior to chlorobutanol feeding. After feeding of chlorobutanol had been discontinued for five and one- half months. the semen values had decreased rapidly. Feeding of chlorobuta- nol was again started. hiring the preceding three month period prior to the start of the second trial. eleven services resulted in only one conception. After feed- ing 1170 grams of chlorobutanol over a four months period in the second trial seven services resulted in three conceptions or 2. 33 services per conception. 811' e " 72-A“ : Sire 72-A was a Holstein-Friesian bull. five years of age, weighing 2330 -Q- pounds. he effect of feeding chlorobutanol on the ascorbic acid content of the plasn and semen. and sperm motility is shown in Table III of the Appendix. Ihis bull received 1080 gm. of chlorobutanol over a period of 149 days. Although Sire '72-A weighed 2200 pounds.it was only able to tol- erate the sedative actioh of 10 grams of chloro‘mtanol per day for 38 days. I'ive grams of chlorobutanol per day was not sufficient to maintain a plasm ascorbic acid level at a sufficiently high plans to produce viable semen. An average dosage equivalent to about seven grams per day appeared satisfactory. Although the blood plasma level reached a peak of 0.5 mg. per cent and the motility reached normal values during the 149 day feeding period, it was never possible to get the ascorbic acid level of the semen up to 5 mg. per amt. Daring the three months prior to chlorobutanol administration the breeding history showed 6. 66 services per conception (17 services and 3 con- captions). while during the first fcur months of trsahnent the number of services per conception was 2.60 (13 services and 5 conceptions). 'Ihis data. is presmted in Table XIII of the Appendix. Sire "1601": 'ihis sire was a two-year-old Holstein-Friesian bull weighing 1600 pounds. It had a normal amount of ascorbic acid in the semen prior to the feeding of the chlorobutanol. Even though this situation existed the sper- matozoa sealed to be of low fecundity as the sire required more services per conception than normal. Data with reference to the blood and semen ascorbic values and motility of the sperm is found in Table XIV of the Appendix. Table XV in the Appendix presents data on the mmber of services per -43- conception. hiring the month prior to treatment the ratio of services to conception was 3.00 (6 services and 2 conceptions). IThe feeding of chloro- butanol for one month rediced this ratio to 1.36 (13 services and 11 con- ceptions) over a period of four months. Sire “morthorn': his sire was a five-year-old bull weigiing 2100 pounds which was sterile at the time the feeding of chlorobutanol was started. All cows bred to this sire for a period of seva'el months had failed to conceive. The bull was brougit to the miry Emerimsnt Station barn for study. The semen was of poor quality as evidenced by the short time liveability of the sperm. nta in Table XVI of the Appendix show that after chlorobutanol ind been fed for some time. the semen did not show any marked change in ascorbic acid content. The sperm. however. showed marked improvement in liveability. Bo Opportunity presented itself to test the fertility of this sire before it had been disposed of to make room for other animals. Table XVII in the Appendix presents data on the per cent of abnormal spermatozoa in the semen of this bull. The semen had 13 per cent abnormal sperm a month after chlorobutanol feeding had started and nien per cent when feeding was discontinued. Two months after the feeding was discontimed the abnormal sperms were back to 12 per cent. At all times a majority of the abnormal sperm had bent bodies and coiled tails. Sire “Wells“ : The Wells" bull was a Jersey three-year-old and weighed 1000 pounds. hiring the three months prior to treatment the number of services per con- ception for this animal was 5.0 (10 services and 2 conceptions). hiring the two months after starting chlorobutanol. the conception rate was 1.6 -44.. (3 services and 2 conceptions). See Table III in the Appendix. The plane ascorbic acid values for plasma and semen were not changed significantly during the first month of chlorobutanol administration. hta were not collected on this animal from June 6 to July 1. Chlorobutanol feeding was started June 26. he plasma ascorbic acid values became markedly higher. Semen ascorbic acid values did not show an increase until July 15. mlorobutanol was discontinued August 17. he plasma and semen ascorbic acid values remained within the normal range until the bull was sold. here was an improvement in sperm motility of the fresh sample and at 24 hours associated with increassdsemen ascorbic acid as shown in Table XVIII of the Appendix. Sire "M20": Sire 13-52:: was a four-year old bull which weighed 2000 pounds. he ascorbic acid content of the blood plasma and the semen data are presented in Table III of the Appendix. his sire had been out of service for several months prior to the time this study was begun. The spermatozoa were all non- motile. Heavy (bass of chlorobutanol were administered orally by capsule in orda' to stimulate the rapid synthesis of ascorbic acid. Chlorobutanol feeding was started June 20. he feeding of 170 grams of chlorobutanol over a 9-day period resulted in muscular incoordination. Although the blood plasma ascorbic acid values attained in four weeks time were as high as those in the other sires used in the experiment it was impossible to bring the semen values into a normal range. Most of the sammes were seminal fluid without sperm. considerable amount of cellular debris and crystals were noticed in the seminal fluid from time to time and on one occasion the fluid was dark brom in color. he feeding of 5,000,000 1.0. of vitamin A in the form .- shark liver -45- oil had no effect on the plasma and semen ascorbic acid values. The pH values on most samples of seminal fluid or semen were higher than found on normal samples of semen. dire “morn": This bull was a seven-year-old Jersey weighing 1100 pounds. mta on the blood and semen ascorbic acid values and sperm motility are found in Table XXII of the Appendix. This sire was able to tolerate a lO-gram dose of chlorobutanol for 23 consecutive days before developing muscular incoordination. A total of 810 grams of chlorobutanol was fed over a 61 day period. The problan with this sire was to improve the fecundity of the sperm. men the sire was placed on the trial the semen as normal as to span motility for fresh samles, at 24-hour and 48-hours of age. Although this was true the conception rate for the three months period prior to the experiment was 4.33 (13 services and 3 conceptions). Daring the three month period following the 61 day feeding trial the conception rate was 1.43 (7 services and 5 conceptions). Data for concep tion rates are found in Table XXII of the Appemdix. Sire 'Ladd": Results of feeding chlorobutanol to sire 'Ladd" a six-year-old Jersey bull weighing 810 pounds are tabulated in Table XXIV of the Appendix. The feeding of 425 grans of chlorobutanol over a period of 85 days resulted in an increase in per cent of motility over the entire 72 hours evaluation period during the first two months and then declined during the rest of the chlorobutanol feeding period. -46- hta in Thble m of the Appendix show. that the breeding efficiency of this sire was not improved to any extent. Before treatment flee number of scvices per conception was 9.00 (9 services and 1 conception). After treatment it was 7.5 services per concep tion (15 services 2 conceptions). These negative results were probably the to the presence of 30 per cent ab- normal sperm and consida‘able mount of cellular debris in the semen. Sire "Rex": Rex. a five-year-old Guernsey sire weighing 1400 pounds was treated on the owner's farm. The only record that could be obtained during the war time travel restrictions was improvement in breeding efficiency. Chloro- butanol. was fed at the rate of five grams per day for 60 days. The results on breeding efficiency are as follows: Period Treatment gervices ConceLti on Services per Conception 3 months previous None 10 3 3.33 3 months following 60 days 8 6 1.33 Sire “Maxim“ : This seven-year-old Guernsq bull weigaing 1500 pounds was fed chloro- butanol at the rate of five grams per day for 60 days. This sire was also treated at the owner 's farm. The results on breeding efficiency are as follows: Period Treatment Services Conception Services per Goncgption 3 months previous None 13 6 ‘ 2.60 3 months following 60 days 10 7 1.45 -47- measurigg the Improvement in Fertility Services per conception for the ten experimental periods using nine sires are aimarised in Table 17. These sires required an average of 5.21 (2.6 to 11.0) services for each conception prior to the oral administration of chlorobutanol. A sire with 2.60 services per conception might be con- sidered a border-line case of low breeding efficiency. Table IV. SUMMARY OF THE USE OF CHLCBOBUTANOL ON THE BREEDING EFFICIENCY 0F SIRES. .: 3 I T 8 : llams of Sire3mys Fed 3Grams of3Services per3Period 38ervices8Period 3Chlorobutan0130hloro- zconception 3Before 3Per Con-3After 3 3butanol zBefore Iced-3(Weeks)3ception 3(leeks) 3 3red xing 3 3After 3 3 3 3 3 Weeding 3 3 3 3 3 3 3 1. Patty 3 42 3 120 3 6.33 3 28 3 1.60 3 6 2. " 3 102 3 490 3 not used 3 - 3 - 3 - 3. 1645 3 55 3 275 3 3.80 3 44 3 2.60 3 20 4. 1645 (2d: 3 3 3 3 3 period 3 158 31170 3 11.00 3 12 3 2.33 3 16 6. 72-A 3 149 3 1080 3 5.66 3 12 3 2.60 3 l6 6. 1601 3 28 3 190 3 3.00 3 4 3 1.36 3 4 7. Wells 3 60 3 390 3 5.00 3 12 3 1.50 3 6 6. Thorns 3 61 3 810 3 4.33 3 12 3 1.40 3 12 9. Ladd 3 85 3 425 3 9.00 3 12 3 7.50 3 12 10. Rex 3 60 3 300 3 3.33 3 l2 3 1.33 3 12 11. Maria 3 60 3 300 3 2.60 3 12 3 1.45 3 12 3 3 3 3 3 3 Total 3 860 3 5840 3 52.06 3 160 3 23.57 3 118 Av. : 78.1 3 530.9 3 5.21* 3 16.0’ : 2.36‘ 3 11.8‘ *AveragLealculated only on the trials where the sires were used. After chlorobutanol had been fed the average number of services per conception was reduced to 2. 36 (1.33 to 7.60). The sire having 7.50 services per concqtion did not respond to chlorobutanol treatment. m a group basis 81 per cent of the sires improved in fertility of the sperm after chloro- butanol was fed. The average number of days that chlorobutanol was fed was 78.1 (42-158) and the average number of grams of the drug that was fed was -48- 530.! (190-1170) grams. Ho ill effects wa'e observed from feeding the chlorobutanol even for the longest period of time. Nature of Improvement in Ferti lit; Therapy for poor fertility in sires by the use of oral administra- tion of chlorobutanol deve10ped with a definite pattern. Data covering this phase of the proJ set is found in Table V. be improvanent was first noticed in increased blood plasma ascorbic acid values. The average days required to reach maximum levels of plasma ascorbic acid for the group of sires studied was 38.7 days. The range was Ron 2) to 60 days. fresh semen samples returned to a 75 per cent progessive motility in 60.4 days for his group. A wide variation also occurred in this category. The most rapid response occurred in fifteen days compared to 128 days for the bull showing slow recovery. TABLE V. LENGTH OF TIME RMUIRED TO OBTAIN IMPROVBAENT IN BLOOD AND SHEEN VALUES AFTER FEEDING CHLOROBUTANOL 3 3 3 3 Mto Obtain 3 3 Name of Sire 3 Age 3 Maximum 3 Maximum 3rresh Samp1e3lmprovement in 3 Yrs. 3 Blood 3 Semen 375 % Motil- 3Conception 3 3 Ascorbic: Ascorbic3ity mate 3 3 Values 3 Values 3 Ihys 3 3 3 3 3 3 Patty 3 2 3 2O 3 27 3 54 3 Yes Patty (2d) 3 3 3 60 3 32 3 60 3 Not used 166 3 3 3 24 3 115 : so : Yes 1645 (as) 3 4 3 2:5 : 127 : es : Yes 72-A 3 5 3 24 3 143 3 128 3 Yes 1601 3 2 3 36 3 36 3 36 3 Yes mar thorn 3 5 3 60 3 67 3 60 3 Not used lells 3 3 3 60 3 75 3 60 3 Yes 13-520 3 4 3 32 3 121 3 Failed 3 No Thorns 3 7 3 48 3 55 3 l5 3 Yes Ladd : 6 3 - 3 - 3 48 3 No Box 3 5 3 - 3 - 3 - 3 Yes Maxim 3 7 3 - 3 - 3 - 3 Yes T0681 8 56 8 337 3 798 2 60.4 2 Yon . 81 fl Av"I 3 4.3 3 38.7 3 79.8 3 60.4 3 I"Average calculated only on number of sires for which there was data -49.. Maximum ascorbic acid values developed much more slowly in semen than in the blood. The average length of time required to reach the highest values in the semen was 79.8 days. Here agin. the variation in time was from 27 to 143 days. The sire showing the slowest recovery in per cent of sperm motility required file highest number of days to attain the maxim amount of plasma ascorbic acid in the semen. Furthermore. the sire that reached the maximm blood values in the shortest length of time also attained the maxim semen values in the shortest length of time. Administration of Large Doses of Onlorobutanol Large does of chlorobutanol were adninistered in an attempt to re- duce the time required to bring about recovery of fertility. Sires studied as reported in Tables VIII. XII. XVI. XVIII and XII of the Appendix received doses of more than five grams per day for various lengths of time. Individual sires showed considerable variation in their ability to tolerate massive dosages of chlorobu tanol. The Patty bull (Table VIII in the Appendix) withstood 20 grams per day for nine days without deveIOping muscular incoordination. This period was followed by four 30 gram doses in capsules over a period of eight days. At the end of this time pronounced incoordination of the muscles deve10ped. With Sire 72-A (Appendix Table III) ten gram‘doses were fed for 38 days. then five grams for 65 days. Following this period this sire was again fed a ten gram dose and in nine days time developed incoordination of the legs. m Shorthorn bull (Table xv: in the Appendix) was fed a ten gram dose for 21 days before evidence of muscle incoordination developed. The Wells bull withstood ten grams per day for ten days, however, the heavy -50- dose equivalent to 15 grams per day fed at the rate of 30 grams every otha' day produced pronounced muscle incoordination in eigit days time. With the 13-520 bull (Appendix Table m) 20 and so gram doses fed by capsule pro- duced a positive incoordination in nine and eight days. respectively. Blood plasna ascorbic acid values increased more rapidly following the massive dose treatment than when small doses were administered. None of the sires exhibited an ill effect after the large doses were discontinued and normal doses of chlorobutanol were again fed. gorpholog‘of the §perm Pour sires were studied with refa'ence to the morphoIOgy of me spermatozoa during a part or the entire period of the experiment. Table II of the Appendix shows the history of the Patty bull. This sire improved rapidly in the appearance of the spermatozoa. After the therapy was discontinued the number of abnormal spernntozoa again increased. In four and one-half months of treatment the abnormal spermatozoa drapped from 76 per cent to 21 per cent and then in two and one-half months without treatment returned to 42 per cent. Tables NH and XI of the Appendix present the data on the 'Shorthorn" and "Wells" bulls. In both of these animals the per cent of abnormal sper— matozoa was not above the average for high quality semen. me sperms of neither one of these two bulls showed any improvemait from a morphological standpoint. The Ladd bull ahd 30 per cent of abnormal sperm after feeding had been in progress for a period of a month. This may explain in part the reason why this sire did not improve with reference to fertility of the sperms. .. 51 .. Chlorobutanol Administration to Cows Scape of Study In this part of the experiment 32 cows with breeding difficulties in four different herds were studied. Two of the horde were privately owned. One was Michigan State College herd and the other the Rutger's University herd. All five breeds of dairy cattle are represented in this study. The results are tabulated in Tables XXVI to xxx inclusive of the Appendix and summarized in Table VI. All of me 32 cows had been examined by veterinarians and pro- nounced normal fmm a clinical standpoint with reference to their repro- ductive organs. Fifteen of the 32 cows or 46.9 per cent had been bred to two or more sires. which tended to eliminate the possibility of the lack of fertility caused by the sire rather than the female under study. Effect of Treatment on Milk Producti on In Table XXVI of the Appendix data are presented showing the effect of normal therapy and shock therapy (massive doses) on the rate of milk flow when lactating female bovines were treated. When five gram doses were used no appreciable decline in milk flow was experienced otha' than the normal decrease the to stage of lactation. Prior to the treatment on five milking cows the average prediction per day was 33.5 pounds of milk. hiring treatment the prediction averaged 31.8 pounds of milk. Smock therapy caused a sharp decline in milk flow amounting to slightly over a 50 per cent de- @0880. - 53 - MARY 0F EFFECT Cl" FEEDING CEOROBUTANCL T0 “HARD TO SETTLE" COWS IKBLE VI. Q. Breed Cows m 1w .1 80 Val mm 0 tn 0 I1 e.t mm ”C em SC @r ac 8? m a 9 CO .1... O m: 0 SB 59 a .1. O In 1 m m... . t a e r T mt rm Mm V. m “.0 n 6 m a m: aaammaamgwawwmu NO. 16171411811211... “.0 2*“ .0 m.“ :m: .OPWW .0 .0 .0 .0 z .. m .omw.on.omwauaumwausuauausuau mrm : .0 $ '0 W .. fi .. .0 fi .0 .. .0 g .0 .. Em .o.v.o.e:ueu:uaun.7.4.374.0.4 N 1 . oommmommoaewemm b wm888w 8 213771 F» 1.1.1. .1 8 ' a m e ."'.""" "." r o e r a :‘w..:...0::..:m::w..: .i r m e . .. a .N .iuu.1 .m.m ya .6 ct wwimmMuanmoa4aw w 00.000.990.000:90.0.0.0:00000000 1.1222112111141162 :..::..::::’:::.0 .0 .0 .0 5343333485555 a 54.236353344334414 8058020 55 9 00 mmmwewmmmmmmammmm 1.1.1. 1. 1.1. 1.1.1.1.1.1. OI n n a. an..." M“ Yam I h“ t. at n I.80.8"I'flhfl”"' m... 1m 00 m :‘COmzm::::wm.l.O::: a a” an " 1.1.1 H. .mmm 4443$fl333$628flwmm JJAGbekkbefiaam A 1167 36.5 .0 .9 .0 .U 62 00...... Total Aw. DISCUSSION OF RESULTS Effect on Libido in Bulls Response to the feeding of chlorobutanol took place usually within a week to ten days. The average number of days that chlorobutanol was fed was 41.8. The range was from 13 to 70 days. The most satisfactory dose appeared to be about five grams per day. Some sires tolerated heavier doses than others. The "Spotts" bull withstood seven grams per day for 34 days as shown in Table I. Blood plasma ascorbic acid values increased greatly in sires ”Petty" and 'Pheifer Boy“ as shown in Table mm in the Appendix. These data are in agreement with the results of Bortree, Huffman and mncan (15), (16). Phillips (26) obtained definite response in 'hlow breeding" bulls by the subcutaneous injection of vitamin C. Chlorobutanol may have produced libido stimulating effect in part at least through increased blood plasma ascorbic acid. “his failure of the 13.4 per cent of the bulls to respond to chloro- butanol may have been due to imprOper function of some other endocrine gland. Phillips (14) was of the Opinion that ascorbic acid has a beneficial effect on the pituitary. Turner (46),Peterson and et a1. (43) and Spielman and co- workers (94) associated lack of libido in sires with low thyroid activity. 'i'nere was no difference between Holsteins and Jerseys in the rate of improvement in libido as shown in Table II. The administration of chlorobutanol reduced the average time of service from 45 minutes to three and seven-tenths minutes: Each sire re- quired an average of 233.4 grams of the drug or the equivalent of about one- half pound. -54- Improvement of libido in slow breeding male bovines was attained in 76.6 per cent of the animals used in the experiment. The loss of libido is not entirely a problem that deve10ps with old sires since 53.3 per cent of the bulls reported as slow breeders in this experiment were under two years of age. Age of the sire was the greatest factor which influenced the degree of response. With the bulls two years old or younger, there was a 94 per cent recovery. Only one sire. two year old or younger, failed to recover completely. The partial recovery was called one-half recovery in calculating the percentage. In the older group of bulls the average age was 6.6 years. Four of these bulls recovered normal libi do. while three bulls failed to recover to any noticeable extent. me average recovery for this group was only 56.2 per cent. Effect of Feeding Chlorobutanol on Fertility of Bulls In this study the oral administration of chlorobutanol remlted in improvement in fertility of nine of the eleven sires or an average of 81 per cent as was indicated by the number of services that were required per conception. These results are in close agreement with those of Phillips (26) in which three out of four or 75 per cent of the sires improved in conception rates by the subcutaneOus injection of vitamin C. It was necessary to feed chlorobutanol for a longer period of time in order to improve the fertility of the sperm than to improve the libido. The data show that an average of 78.1 days were required to improve fertility and only 41.8 days to improve libido. The improvement in fertility was in- dicated by the following results: (a) The blood plasma ascorbic acid value reached a maximum in 38.7 days. (b) The fresh samples of semen showed 75 -55- per cent progressive motility within a mean of 60.4 days. It is interesting to note that ascorbic acid content of the blood of the sires took more days to reach a maximum content than for the motility to reach 75 per cent or more for the flesh semen samples. It took on the average 79.8 days for the semen sanples to reach maximum vitamin C values. The 60.4 days required for the semen values to reach 75 per cent progressive motility on the fresh samples after starting chlorobutanol administration is considerably longer than the five we‘ks of treatment with subcutaneous inJections of vitamin C as reported by Hiillips and co-workers (26). The bulls low in fertility as reported by breeders for this study averaged 4.3 years of age. The ages of the two sires that did. not respond to treatment as reported in Table V were four and six years, respectively. The data presented in Table IV show that the bulls which received chlorobutanol were not brought back to the average of 1.56 services per conception as reported by Bowling and associates (1) or 1.52 for bulls under five years as reported by Hilder and co-workers (4). The data in Table VIII show that in six of the ten trials or 60.0 per cent of the sires that were used for breeding had a breeding efficiency of 1.50 services or less per conception after treatment. All of these sires were fed rations that should have maintained fertility. None of these bulls wa'e in poor flesh due to malnutrition when they was reported as having a poor breeding efficiency. It was observed that the animals umally took on somewhat of an added bloom during the period when chlorobutanol was fed. No apparent detrimental effects were observed from the chlorobutanol feeding up to 158 days. These results are in agreement with the work of Lundquist and Phillips (96) who reported that the feeding of chlorobutanol at the rate of five grams per day had no -55- detrimental effect on the synthesis of the vitamin B-complex. This is of interest because chlorobutanol has a snail phenol coefficimt. Chlorobutanol therapy as an aid to correcting abnormal sperms could not be measured since there was only one sire that had a high percentage of abnormal sperm. However, there was an improvement in the morph010gy of the sperm during the time chlorobu tancl was fed in the case of this bull. An interesting observation was that the abnormal sperm with coiled tails in many instances lived longer than the normal sperm. In addition their path of locomotion was reverse to that of morphologically perfect sperms. Effect on Breeding Efficiency in Cows In measuring the value of the oral administration of chlorobutanol to ”hard to settle" cows. only cows that had a clinically normal reproduc- tion tract were used. The basis for using the chlorobutanol was that Phillips and co-workers (24) had reported that the subcutaneous injection of vitamin C resulted in the improvement of 60 per cent of the cows with breeding difficulties. Boy (30) working with humans showed that ascorbic acid administration aided pregnancy in women by keeping up the ascorbic acid content of the corpus luteum. Ihvis and Cole (3) reported favorable results from the subcutaneous injection of vitamin C in the breeding per- forsance of mares. Since Bortree et al. (15). (16) had shown that blood plasma levels could be raised measurably by orally administrating chloro- butanol it appeared likely that the use of this compound might be valuable for overcoming shy breeding in the female bovine. The number of services prior to feeding (the chlorobutanol ranged from one to 14. The breeder who started the cow on chlorobutanol after the first service did so because the cow had a history of shy breeding. Twenty -57- of the 32 cows that had an average of 4.55 services prior to treatment conceived on the first service after treatment. The entire group of 32 cows had an average of 4.84 services per conception before treatment. The feeding of the drug was usually started immediately following the cessation of the heat period and continued for at least 21 days. The average feeding period for the 32 head was 36.5 days. Following chlorobutanol therapy, the 32 cows averaged 2.00 per services per conception. Twenty of the 32 animals or 62.5 conceived on the first service after therapy. The 12 cows which reouired more than one service, averaged 5.33 services before treatment. and 3. 83 services following treatment. It is likely that some of the 32 cows reported in this stucb' would have eventually conceived without treatment. Asdell and associates (103) using 4'? sterile dairy cows reported that the control group eventually had 50 per cent of the cows conceive while those that had been treated with gonadic and gonadotrOpic hormones only 42.9 per cent conceived. In this experiment all the cows treated conceived, which indicated that the treat- ment with chlorohitanol improved the breeding efficiency. Effect of Shock Therapy As shown in Table VI, there were five cows included in this study that were treated with massive or shock doses. Four cows received 20 grams daily for a period of five days and the fifth cow received eight grams for 22 days. These five cows had an average of five serVices prior to treat- ment. All of the cows conceived on the first service except one, following the shock treatment. This cow required only two services. -58- Five of the 32 cows were'fed three grams of chlorobutanol per day instead of five grams. The average number of services before treatment was 3.4. Following treatment all the cows conceived on the first service except one which required two services. Effect on Milk Production Milk production was affected only slightly by the normal dosage of three to five grams of chlorobutanol per day as shown in Table XXVIII in the Appendix. Shock or massive doses of 20 grams of chlorobutanol per day reduced milk flow 56.5 per cmt. This may have been due to the reduction in appetite associated with the sedative effect of chlorobutanol. Whether or not the action of the chlorobutanol is a primary or secondary one was undetermined. The ascorbic acid may provide the correct media for the enzyme which breaks down the cell wall of the ovum permitting the sperm to enter. The return of the treated cows from 4.84 services before treatment to an average of 2.00 services per conception after treatment may be inter- preted as returning the cows to a normal condition. This statement is based on the report of Bowling and collaborators (1) who found that the averages of all services for the lifetime of a cow to be 2.02. - 59 - SUMMARY AND CONCLUSIONS Fifteen slow breeding bulls were fed chlorobutanol orally. The libido of the bulls improved in 76.6 per cent of the cases. The average time required to serve a cow was reduced from 45 minutes to 3.7 minutes. There was no difference between Jersey and Holstein-Friesian sires in response to chlorobutanol administration from the standpoint of libido. Sires two years or under had an average recovery of 94 per cent com- pared to 56.2 per cent recovery for a group of sires that averaged 6.6 years of age. Plasma ascorbic acid values showed a peak 38.? days (average) after starting chlorobutanol feeding. The maximum semen ascorbic acid value. however. was mt reached until 179.8 days (average) following chloro- bu tanol admini stratio n. The feeding of chlorobutanol to nine sires with histories of low breed- ing efficiency which were in service after treatment resulted in the redaction of the number of services per conception from 5.21 to 2.36. This treatment improved the breeding efficiency of 81 per cent of these bulls. In this study an average of 60.4 days was required for the fresh semen to attain ’75 per cent or more of progressive motility after starting - chlorobutanol feeding. No ill effects were observed from feeding a total of 10 grams of chloro- butanol over a period of 158 days, or an average of five grams per 100;) 10. 11. -50.. pounds of body weight per day. Only two sires. however, were able to withstand doses of 20 grams per day over more than a three week period without deve10ping muscle incoordination. Sires used in the improved breeding efficiency and libido studies were fed average daily doses of 6.8 and 6.1 grams of chlorobutanol, respec- tively. These doses were equal to 4.9 and 5.0 grams per 1000 pounds of live weight, respectively. The most satisfactory daily cbsage for feeding periods of 60 to 150 days appears to be five to seven grams. Thirty-two shy breeding cows with an average of 4.84 previous services were used in this investigation. Twanty of these cows conceived on the first service following the feeding of five grams of chlorobutanol per day for about 21 days. Five of the 12 cows which did not conceive on the first service were fed large doses of chlorobutanol (up to 20 grams per day) until they showed incoordination. Four of these five cows conceived on the first service following this treatment while the fifth cow conceived on the second service. Twenty-four of the 32 cows used in this study received an average daily dose of five grams of chlorobutanol per 1000 pounds live weight. These of three and four grams per day appeared Just as effective as five gram doses. All 32 cows used in this study conceived after treatment with chlorobutanol with an average of two services per conception. Five lactating cows which were fed five grams of chlorobutanol per day declined only five per cent in milk production. Milk production de- creased 56.5 per cent in two cows fed 20 grams of chlorobutanol per cow per day. (1) (2) (s) (4) (5) (6) (7) (a) (9) (10) (11) (12) -51.. LITERATURE CITED Bowling, G. A. 194). Age as a factor influencing breeding efficiency in a dairy herd. J. of Dairy Sci. 28: 1171-1176. Baker, A. L. and Qiensenberry, J. R. 1944. Fertility of range beef cattle. J. of Animal Sci. 3: 78—87. ‘mvies, G, K. and Cole. 0. L. 1943. The relation of ascorbic acid to breeding performance of horses. J. of Animal Science 2: 53-58. Heider, R. 1., Fohrman. M. H. and Graves, 3. R. 1944. Relation of various factors to the breeding efficiency of dairy animals and to the sex ratio of the offspring. J. of miry Sci. 27: 981-992. Baillips, P. H. . 1943. Sterility, some causes, and handling. Jersey Bulletin 62: 1203. . Head, 8. 1., Gregory, P. W. and Regan. I. M. 1946. Deleterious recessive genes in dairy bulls selected at random. J. Dgiry Sci. 29: 557-558. Quinlan, J. and noux, L. I... 1936. Researches into sterility of cows in South Africa. 0nderstepoort J. of Vet. Sci. and Animal Ind. 6: 719-773. Rhoad, A. O. 1944. Rate of conception in beef cows pasture—bred during a con- trolled breeding season. J. of Animal Sci. 3: 153-158. Accame, F., Mange. L. and Miller, J. C. 1944. Effect of reduced barometric pressure on rem semen. J. of Animal Sci. 3: 431. Salisbury. Ge '0 1943. Feed, climate, affect bull fertility. Jersey Billetin 62: 1194. D'b. R. 3., Andrews. F. N. and Hilton. J. H. 1942. Seasonal variation in semen quality of the dairy bull. J. of Dairy Sci. 25: 815-825. Swanson, E. W. and Herman, H. A. 19%.. Variation in bull semen and their relation to fertility. J. of miry Sci. 24: 321-331. -52.. (13) Baltzer, A. C. 194). Causes for cow removal in Michigan herds under test in miry Herd Improvement Associations. Mich. State 0011. Quar. Bull. 22: 147-153. (14) Phillips, P. 3., Lardy, H. 11., Beizer. n. E. and Rupel, I. w. 194). gaerm stimulation in the bull through the subcutanious administration of ascorbic acid. J. of Dairy Sci. 28: 873-878. (15) Bortree, A. I... Huffman, C. F. and mncan, C. W. 1942. Normal variations in the amount of ascorbic acid in the blood of dairy cattle. J. of Dairy Sci. 25: 983-990. (16) Bortree, A. I... Huffman, C. F. and Dincan, C. W. 1943. Ascorbic acid stimlation in the blood plasma of dairy cattle promced by the ingestion of chlorobutanol. J. of Dairy Sci. 26: 553-562. (17) Sutton, r. s., Kaeser, H. m. and Hausard, s. I... 1942. Some factors affecting the synthesis of ascorbic acid in the Albino rat. J. of Biol. mom. 144: 183-191. (18) Stone, c. P. 1939. Capulatory activity in adult male rats following castra- tion and injections of testosterone prOpionate. Endocrinology 24: 165-174. (19) Mindlin, R. L. and Butler, A. M. 1937. The determination of ascorbic acid in plasma: a macro- method and micromethod. J. of Biol. Chem. 122: 673-686. (20) Walbach, S. B. and Bessy, 0. A. 1942. Tissue changes in vitamin deficiencies. Physiol. Kev. 22: 233-289. (21) Snernov-Ugyunow, D, N. and Laptev, M. A. 1945. The feeding of stud bulls (bring mating season. Probl. Zhirotu 2: 64-72 cited by Jones, I. R.. Ibughtery, R. W, and Hoag, J. R. J. of Dairy Sci. 1945 28: 311-320. (22) Ginningham, I. J. and Hepkirk, c. s. M. 1935. Dietary protein in relation to sterility. New Zeal. J. Sci. and Tochnol. 17: 420-432 cited by Jones, I.P.., Daugherty, R. W. and Hoag, J. R. J. miry Sci. 1945 28: 311-320. (23) Webster, W. M. 1932. Bovine sterility in New Zealand. Austral. Vet. J. 8: 199-222. cited by Jones, I. R., Daugherty, R. l. and Hoag, J. R. J. Dairy Sci. 28: 311-320. (24) (25) (36) (27) (28) (29) (30) (31) (32). (33) (34) (35) -53.. Phillips. 1’. H.. Lardy, H. A., Boyer, P. D. and Werner, G. M. 1941. The relationship of ascorbic acid to reproduction in the cow. J. of Dairy Sci. 24: 153. Walton, 5., Edwards, J. and Hammond, J. 1940. Fertility in farm animals. J. Royal Agric. Soc. of England 100: Part III. Phillips, 1’. H., Lardy, H. A., Heizer, E. E. and Rupel, I. W. 1940. New hepe for the sterile bull. Hoard's Dairyman, Mar. 167. Moore, 0. R. 1941. Glandular physiology and therapy (physiology of the testis). J. Am. Med. Ass'n. 116: 1638-1644. Ibnham, C. R. and Simone, B. T. 1931. Fertility studies in the bull. J. Am. Vet. Med. Ass'n. 78: 658-665. Zittle, C. A. and Zitin, B. 1942. Non-hemin and total iron in bull spermatozoa~ J. of Biol. Chen. 144: 105. Lav. L. 1937. Treatment of habitual abortion with vitamin c. Manon. Med. Wochenschr 84: 1814-16. Nutr. Abs. and Reviews 7: 1088. Landquist, N. S. and Hiillips, P. A. 1945. Some observations on the effect of sulpha drugs on blood plasma ascorbic acid levels in calves. J. of miry Sci. 28: 139-145. Berg, 0. C., Huggins, C. and Hodges, C. V. 1941. Concentration of ascorbic acid and the phosphatases in pecretion of the male genital tract. Am. J. of hysiol. 133: 82-87. Belasco, I. J. and Mulin, J. R. 1940. The effect of vitamins A and C on experimental hyper- tmroidism. J. of Nutrition 20: 577-588. Biskind, G. R. and Glick, D. 1937. The vitamin C in the testis in relation to anatomic and functi anal changes. Arch. of Path. 23: 363-371. Sutton, T. S., Krauss, W. E. and Hausard. S. L. 1940. The effects of vitamin A deficiency on the young male bovine. J. of Dairy Sci. 23: 574. (36) (37) (38) (39) (40) (41) (42) (43) (44) (45) (46) (47) -54.. altmn. T. s. 1941. High quality roughage gains new importance. Jersey Bulletin 60: 433. Umava, C. T'. 1941. Vitamin C in children in Begata. J. of Am. Med. Ass‘n. 116: 2109. Phillips, P. B., Rupel, I. W., Aleson, J. J. and Bohstedt, G. 1938. The effects of an atypical blindness producing ration upon the vitamin K and C content of calf blood. Proc. Amer. Soc. of Animal Pro. p. 320. Sire, B., Theis, R. 11., Harrelson, R. T. 1939. Vitamin interrelationships. 1. Influence of avitaminosis on various tissues and endocrines. J, Biol. Chem. 129: 245. Riddell, W. H. and Whitnah. C. H. 1938. Vitamin C metabolism in the dairy cow. J. of kiry Sci. 21: 121. Wallis. G. C. 1943. Evidence of the synthesis of vitamin C by dairy cows. J. of Dairy Sci. 28:401 Almquist. J. O. and Andrews, 1'. N. 1944. The failure of ascorbic acid to augment equine gonadotrcpin in the rat. J. of Animal Sci. 3: 183-187. Peterson, W. B., Spielman, A.. Pomeroy, B. S. and Boyd, W. L. 1941. Effect of thyroi dectomy upon sexual behavior of the sale bovines. Proc. Soc. of Exp. Biol. and Med. 46: 16-17. Erb. R, E. and Andrews, 1'. N. 1942. Effect of the gonadotropic substance of pregnant mares serum on the blood plasma-ascorbic acid of the bovine. 30: 258-262. DiCio, A. B. and Schteingart, M. 1942. The influence of ascorbic acid on the activity of gonadotropic hormones. Endocrinology 30: 263-264. Turner. C. '0 1943. Sterility in sires and cows. Guern. Breeders Jam. 63: 712-713. Johnson, 8. R. 1943. Studies with swine on rations extremely low in maganese. J. of Animal Science 2: 14. (#9 (49) (50) (51) (52) (53) (54) (55) (56) (57) (58) (59) -55.. Jones, I. B., Daugherty, R. W. and Haag, J. R. 1945. Relation of nutrition to growth and breeding performance in dairy bulls. I. Alfalfa hay rations. J. of Dairy Sci. 28: 311-320. Roberts, Elmer and Dawson, W. M. 1935. Effect of Folwers solution on animals. Bull. 413: 187-202. Univ. of Ill. Holt, E., Albanese. A. A., Shettles, L, B., K3,} di, C. and Wangerin, 13.11. 1942. Studies of eXperimental amino acid deficiency in man. I. Nitrogen balance. Fed. Proc. - Fed. of Amer. Soc. of sap. Bio1. 1: Part II, 116-117. Boyer, P. 1)., Shaw, J. H. and Phillips, P. H. 1942. Studies on managenese deficiency in the rat. J. of Biol. Chem. 143: 417. Lardy, H. A" Boyer, P. B., Slaw, 1942. Manganese for cattle. and Phillips, 1’. A. Board's miryman 87: 558. Kuhlman, A, H. and Gallup, W. D. 1942. Carotene requirements of dairy cattle for conception. J. of Dairy Sci. 25: 688-689. Reid. J. T‘. and Sykes. J. 1'. 194.5. The influence of ascorbic acid on the activity of gonadatropic hormones of guiena pigs. (unpublished data). Biskind, G. R. and Glick. D. 1936. The vitamin C concentration of the corpus luteum with reference to the stage of estrous cycle and pregnancy. J. Biol. Chem. 113: 27-31. Davis, H. P. and Williams, N. K. 1939. Evaluating bovine semen. Proc. Soc. of An. Prod. 232-242. Undeer erg. G. L. K. and Davis, H. P. 194). Longevi ty. fecundi ty. motility and pH of diluted and undiluted semen. Proc. J. of Soc. of An. Prod. 229-235. 'eatherby, E. J., Reece, P. R. and Bartlett, J. W. 1940. The ability of dairy bulls to withstand regular service for artificial insemination during one year. Proc. Soc. of Animal Prod. 224-229. Salisbury, G. W. 1941. Recent research developments in the preservation and handling of bovine semen. The Cornell Veterinarian, 31: 149-159. (60) (61) (62) (63) (64) (65) (66) (67) (68) (69) (70) (71) Nillett, E. L and Salisbury, G. W. 1942. me effect of various dilutors, cooling rate, temperature of storage and some other factors on the liveability of spermatozoa in stored samples of bull semen. Memoir 249, Cornell Univ. 3-4.5. Bortree. A. L. 1941. A study of some of thefactors affecting the amount of ascorbic acid in the blood plasma of cattle. Thesis. Michigan State College. Schaffner. C. S. and Andrews. 1'. N. 1943. The determination of the concentration of spermatozoa in fowl and bull semen. Anatomical Record 86: 99-107. Brown, I. C., Vanlandingham, A. H. and Westley, C. E. Jr. 1941. Oxidized flavor of milk. I. The effect of feeding postassium iodide supplements to dairy cows on the carotene content of the butterfat and the ascorbic content of the milk and the relationship to metal- induced oxidized flavor. J. of miry Sci. 24: 1035-1039. n'ill, V. A. 1943. Interrelation between thyroid function and vitamin metabolism. Physiol. Reviews 23: 355-379. Beck, G. H. and Salisbury, G. W. 1943. Rapid methods for estimating the quality of bull semen. J. of miry Science 26: 483-494. Davis, R. P., Undeererg, G. I. and Williams. N. Y. 1940. The effect of storage temperatures upon certain diaractsr- istics of bovine sane). J. of wiry Sci. 28: 1057-1068. Letard, E. 1936 Artificial ineemimtion in the domestic animals. Vet. Jour. 92: 61. 1935. The physiolog of spermatozoa. Proc. of Animal Prod. 222-2350 Lesley. J. F. 1944. The relationship between spermatozoan motility and the percentage of live spermatozoa and fertilizing capacity of bull semen. J. of Animal Sci. 3: 433. Ely, R. B., Herman, H. A. and Winchester, C. F. 1942. Studies of respiration rate of dairy bull spermatozoa. Univ. of Mo. “es. Bull. 353: 1-21. Lasley, J. Land Bogart, R. 1944. A comparative study of epididymal and ejaculated sperma- tozoa of the boar. J. of Animal Sci. 3: 360-370. (72) (73) (74) (75) (76) (77) (78) (79) (80) (81) (92) (a 3) -67- Bartlett, J. l. aid Perry. I. J. 1939. Lessons learned from eighteen months experience with c00perative artificial breeding of dairy cattle in New Jersey. Proc. Amer. Soc. Animal Prod. 32nd meeting. Albrecht. I. A. 1945. Discrimination in food selection of animals. The Scientific Monthly 60: 347. Pankevic. A. A. 1945. The effect of the protein level of the diet on the sperm of breeding bulls. Feeding of farm animals and fodder prod. 79-93. Pub. by Lenin Acad. Agric. Sci. Moscow 1940. Nutr. Abst. and Reviews 14, 3: 576. Anderson, J. 1941. Further investigations on the semen of the bull. The Veterinary Record 53: 197.205. Hammond, J. 1921. Further observations on the factors controlling fertility and foetal atrOpy. J. Agric. Sci. 11: 337-366. Zittle, C. A. and O'dell, R. A. 1941. Chemical studies of bull spermatozoa. J. of Biol. Chem. 140: 899-907. Daugherty, R. W. and Ewalt, H. P. 1941. Semen studies in the 3:211. Amer. J. of Vet. Res. 11. 419-426. MacLeod, J. 1941. The metabolism of human spermatozoa. Am. J. of Physiol. 132: 193. Kelley, P. L. 1940. How should we manage the herd sire. Roard'. Dairyman 85:40. Willett, E. L., miller, H. K. and Salisbury, G. l. 194). Preservation of bovine spermatozoa in yolk-phosphate dilumt and field results from its use. The Cornell Veterinarian 30: 507. Phillips, R. I" Knapp, B. Jr., Hanstra. L. C. and Eaton, O. N. 1943. Season variation in semen of bulls. Amer. J. Vet. Res. 4: 115-119. Laptev. M. A. 1945. Thseffect of feeding on the sexual activity and sperm of bulls. Feeding Farm Animals and Fodder Prod. 67-78. Pub. by Lenin Acad. Agric. Sci. Nutri. abs. and Reviews 14: 3: 5'76. (84) (85) (86) (87) (88) (89) (90) (91) (92) (93) (94) (95) -58- Pinetean. M. 1939. The aphrodisiac properties of ergot of rye. J. Amer. Vet. Med. 95: 511. Timin, X. S. and Perelurien, M. I. 1944. The influence of wheat germ on sperm production of bulls. Akod. Sel. Skokhoz. Newk. 6: 17-21. Vet. Bull. 14:278. Her-Ike. L. A. 1935. Results of feeding sprouted cats to correct sterility in cattle and swine. J. of Agric. Res. 51: 51-59. Gullickson, L. 3., Palmer, W. L. B. and Olson, F. C. 1944. Vitamin E in the nutrition of cattle. Jour. of Dairy Sci. 27: 634. Salisbury, G. W. 1944. A controlled experimmt in feeding wheat germ oil as a supplement to the normal ration of bulls used for artificial insemination. J. of miry Sci. 27: 551-562. Donham, C. B., Simms, B. T. and Shaw, J. N. 1931. Fertility studies in the bull. II. The relation of miscroscopic findings in semen to its fertility. J. Am. Vet. Assoc. 78: 665-680. Salisbury, G. W. and Mercier, E. 1945. The reliability of estimates of the proportion of morphologically abnormal spermatozoa in bull semen. J. of Animal Sci. 4: 174-177. Swanson, E. W. and Herman, H. A. 1944. Seasonal variation in semen quality of some Missouri dairy bulls. J. of Dairy Sci. 27: 303-310. Erb, H. B., Andrews, F. N., Bullard. J. F. and Hilton, J. H. 1944. A technique for the simultaneous measurement of smen quality and testis histolOgy in vitamin A studies of the dairy bull. J. of Dairy Sci. 27: 769-772. Swanson. E. W. and Herman, H. A. 1944. The correlation between some characteristics of dairy bull semen and conception rats. J. of Dairy Sci. 27: 297. Spielman, A. A., Petersen, N. B., Fitch. J. B. and Pomeroy, B. S. 1945. General appearance, growth, and reproduction of thyroidectomized bovine. J. of miry Sci. 28: 329-337. Guilbert, H. R. 1942. ”Some endocrine relationships in nutritional reproductive failure." (A.review) J. Animal Sci. 1: 5-13. . P v t r e t e l l I. ‘ I t 0 v 7 s . Q 0 I t o ‘ I ‘ l s 4 s . l c s l r. I a s e l c P n l n I s e s P 6 s _ s . O A ‘ V ' w s D f v a . s Q s. s e I f e s 0 U u I s _ J u l s I e n s s s s A s f l n a v o o s f s n n I _ . _ \ O s n s e I {SD-I... w.“ 1 F‘.’ .2. . . . . (96) Lindquist, N. S. and Phillips, P. H. 1945. The effect of chlorobutanol on certain members of the B complex in the rumen and blood plasma ascrobic acid levels. J. of Dairy Sci. 28: 25-28. (97) Moore, L. A. and Cotter, J. W. 1945. The relationship between a low carotene intake and urinary excretion of ascorbic acid in dairy cattle. J. of miry Sci. 28: 495-506. (98) Kimble, M. s. and Gordon, s. s. 1939. The importance of riboflavin and ascorbic acid in the utilization of vitamin A. J. of Biol. Chem. 128: 111. (99) McHenry, E. W” Reedman, E. J. and Sheppard, M. 1938. Physiological proper ties of ascorbic acid. Biochem. J. 32: 1302-1304. (100) Musulin, R. R., Tully, 3. H. 3d., Longneckar, H. E. and King, C. G. 1939. Vitamin C synthesis and excretion by the rat. J. Biol. Chem. 129: 437. (101) Smythe, c. v. and King, 6. G. 1942. A study of ascorbic acid synthesis by animal tissue in vitro. J. Biol. Chem. 142: 529—541. (102) Longnecker, H. E., Musulin, R. B., Tully, R. H. 3d. and King, C. G. 1939. An acceleration of vitamin C mthesis and excretion by feeding known organic compounds to rats. J. Biol. Chem. 129: 445-453. (103) Asdell, s. 1., Fincher, M. G., Smith, s. E. and Elliott, r. I. 1942. A controlled attempt to restore fertility in dairy cattle by treatment with gonadic and gonadotropic hormones. Cornell University Memoir 243, 1-24. APPENDI X .. 70.. TABLE VII. macs or FEEDING CHLCROBUTANCL 10 THE "PATTY" BULL (JERSEY) mte :Ascorbicicid per 190 m1.: Sperm Motility : Chlorobutanol : Blood Plasma : Semen : Evaluation : Treatment : : : Fresh Sample : : mg. ' mg. 3 (1) : 3 3 3 3 2/28/41 : 0.22 : 10.47 : + : 3/2/ 41 : : : :Started feeding : : : :5 gm. per day 3/7/41 : 0.28 : 4.80 : + : : : : : 3/14/41 : 0.412 : 8. 83 : + + : 3 3 3 3 3/22/41 : 0.510 : 7.73 : 1- + + : 3 3 3 3 3/29/41 : 0.270 : 8.89 : + + : 3 3 3 3 4/4/41 : 0.397 : 7.52 : :- + + : 3 3 3 3 4/12/41 : 0.350 : 8.77 : + 1- + :Discontinued , g : :chlorohutanol 4/25/41 : 0.079 : 7.71 : + + e 1. : 3 3 3 3 5/3/41 : 0.123 : 5.27 : + t t t : 3 3 3 3 5/17/41 : 0.246 : 5.59 : + :- + + : 3 3 3 3 5/31/41 : 0.187 : 6.41 : + -t t + : 7/14/41 : 0.291 : 6.90 : 4. + + + : 3 3 3 3 9/22/41 3 0.238 : 8.04 : + 1» + + 3 3 3 3 3 (1) 1' : 20-40 per cent motility: :- + = 40-60 per cent motility; .- *- 4 : 60-75 per cent motility: + + i- ? : 75 per cent motility. -71- TABLE VIII. PLAsrrA ASCCFLBIC ACID AND seam: DATA FOR THE "mm?" BULL (SECOND TRIAL) 3 3 3 3 3 Date : Ascorbic Acid :Sperm : Vol. 3 Semen :_ Per Cent Motility :_per 100 m1. :Conc. : 0.0. 3 pH : Frash : 24 hr. : 48 hr.: 72 hr. : Blood : Semen : : : : : : : : mg. : mg. :lcc : : : : : : 3 3 3 3 3 3 3 3 3 3/27/42: 0.100 : 0.572 : - : 3.0 : - : 40 : 20 : o : 0 4/3/42 : 0.200 : 3.68 : - : 3.0 : - . 10 : 0 : 0 : 0 4/17/42: 0.244 : 1.456 : - : 2.0 : -' ' 70 : 20 : 0 : 0 4/21/42: 0.190 : 2.083 : - : 3.5 : 7.12 : 50 : 0 : o : 0 4/23/42: 0.150 : 1.766 : 712 : 3.0 : - : 30 : 10 : 0 : 0 4/28/42: 0.098 : 1.953 : 432 : 3.0 : 7.08 : 40 : 20 : 10 : 10 5/6/42 : 0.235 : 4.269 : 160 : 3.0 : 7.30 : 40 : 5 : 0 : 0 5/7/42 : Started feeding 10 gr. chlorobutanol every other day 5/7/42 : 0.216 : 4.685 : 176 : 3.5 : 7.14 : 40 : 20 : 10 : 0 5/12/42: 0.235 : 5.280 : 288 : 4.5 : 7.40 : 40 : 10 : o : 0 5/19/42: 0.350 : 5.346 : 260 : 6.5 : 7.12 : 50 : 20 : 10 : 0 5/26/42: 0.329 : 6.565 : 808 : 3.0 : 7.28 : 60 : 5 : 0 : 0 5/28/42: Discontinued 10 gr. everyother day after feeding 100 gr. 6/2/42 : 0.312 : 6.747 : - : 5.0 : 6.90 : 40 : 10 : O : 0 From 5/29 to 6/6 fed 4 20-gr. doses 6/9/42 : 0.236 : 9.105 : 872 : 3.0 : 6.98 : 30 : 15 : 0 : 0 6/9 to 6/17 fed by capsule four 30-gr. doses. In-coordination 6/18/42: 0.360 : 4.120 : - : 5.8 : 7.06 : 70 : 20 : 0 : 0 6/26/42: Started feeding 10 gr. doses usually every other day 7/1/42 : 0.321 : 3.500 : - : 4.5 : 7.40 : 0 : 0 : 0 : 0 7/7/42 : 0.573 : 1.941 : — : 6.0 : 6.48 : 75 : 10 : 0 : 0 7/15/42: 0.288 : 5.869 : - : 4.0 : 6.88 : 65 : 3 : 0 : 0 7/22/42: 0.430 : 4.927 : - : 5.2 : 7.02 : 80 : 35 : 15 : 0 7/28/42: 0.494 : 6.797 : — : 3.2 : 6.88 : 70 : 50 : 30 : 20 7/21/42: 0.316: - : - : — : .. : 70 : 40 : 20 : 5 8/4/42 : 0.369 : 7.870 : - : 5.0 : 6.86 : 80 : 40 : 40 : 10 8/10/42: 0.371 : 4.059 : - : 3.2 : - : 70 : 55 : 35 : 5 8/17/42: 0.373 : 6.296 : - : 1.7 : 6.9 : 45 : 20 : 20 : 0 8/17/42: Fed 190 gr. since 6/26 - discontinued feeding 8/24/42: 0.419 : 4.370 : - : 2.5 : 7.0 : 35 : 35 : 30 : 25 8/21/42: 0.405 : 4.531 : - : 3.0 : 6.8 : 70 : 55 : 15 : 0 9/4/42 : 0.182 : 5.830 : - : 4.0 : 7.0 : 60 : 46 : 15 : 0 9/12/42: 0.252 : 5.790 : - : 4.4 : - : 70 : 25 : 1 : 1 9/22/42: 0.323 : 6.059 : - : 1.7 : - : 75 : 25 : 15 : 10 9/20/42: 0.319 : 5.479 : - : 1.3 : - : 15 : 15 : 10 : 1 10/15/42: 0.340 : 3.134 : - : 2.6 : - : 50 : 25 : 20 : 1 10/20/42: 0.362 : 4.226 : - : 4.0 : - : 80 : 20 : 10 : 0 10/27/42: 0.272 : 2.537 : : 1.5 : 6.9 : 85 : 10 : 0 : 0 11/3/42 : - : 2.070 : - : 1.6 : 7.12 : 55 : 10 : 0 : 0 11/10/42: 0.403 a 3.948 : - : 2.0 : 7.25 : 45 : 35 : 15 : 5 11/17/42: 0.374 : 3.655 : - : 3.2 : - : 70 : 60 : 25 : 15 11/24/42: 0.287 : 2.781 : - : 2.8 : - : 65 : 30 : 0 : 0 continued on next page TABLE VIII. - continued : Ascorbic Aci d mte : Sperm : Vol. : Semen : Per Cent Motility :ier 100 ml. 3 Cone. : c.c. : pH :B‘resh: 24 hr.:48 hr.: 72 hr. : Blood 3 Semen 3 3 3 : 3 3 3 : mg. : mg. : m/cc : : : : : : g g or 3 3 3 3 3 3 12/8/42 : 0.287 : 3.207 : - : 2.6 : - : 35 : 30 : 12 : 0 12/15/42: 0.366 : 3.570 : - : 3.2 : 7.12 : 65 : 45 : 0 : 0 12/22/42: - : 4.55 : - : 6.7 : - : 45 : 5 : o : 0 12/29/42: 0.327 : 8.415 : 1.584 : 4.0 : - : 40 : 20 : 5 : 0 1/5/43 : - : 4.695 : - : 2.6 : - : 55 : 45 : 15 : 0 1/14/43: 0.275 : 3.123 : - z 3.4 : - : 65 : 40 : 0 : 0 2/24/43: 0.399 : 2.804 : - : 3.8 : - : 70 : 40 : 15 : 10 3/11/43: 0.323 : 4.418 : - : - : - : 45 : 40 : 25 : 15 3/18/43: 0.259 : 2.501 : - : 5.4 : - : 60 : 35 : 15 : 5 4/27/43: 0.544 : 4.552 : - : 4.8 : - : 20 ; 10 : 0 : 0 : 3 3 : : : . 3 2 TABLE III. PPR CENT or ABNCEMAL SPEDMATOZCA :1: 5mm: 01“ "PATTY" BULL IliRING SECOND TRIAL mte : Per Cent Abnormal Sperm : Comments 3 3 4/12/42: 76 : Majority coiled tails 3 3 5/19/42: 62 : “ N " 6/26/42: 42 : " n n 3 3 7/26/42: 45 : N n n 3 3 8/24/42: 21 : .. w n 3 3 11/10/42: 42 : u a n -73- TABLE 1. msCT or mmomTANCL mum: T0 BULL “1645' (BCLST'EIN). iAscorbic Acid per 100 m1. 3 nte :spern :Chlorobutanol treat-set : Blood : Semen :Motili :y: : lg. : . : Trash : : "L : i T : 1/17/41 : 0.28 : 0.88 : None : - 1/39/41 : - : - : :Started feeding 5 grm.per day 1/27/41 : 0.30 : 0.288 : + :Continuing 2/5/41 : 0.43 : 10.22 : + : I 2/8/41 : 0.43 : 8.98 : + + : " 2/10/41 : : : :Discontinued 2/15/41 : 0.52 : 5.52 : + + 4 : 2/22/41 : 0.31 : 6.72 : + + : 35/1/71 : 0.36 : 8.3) : + : 3/17 41 : A : : :Started feeding 5 grl.per day 35/22/41 3 0.389 : 6.22 : + + 4- :Contimed 35/29/41 : 0.495 : 4.08 : + «t + : ' 4/5/41 : 0.494 : 9.630 : + + + : I 412/41 3 0.435 : 7.77 :+ + i- + :Discontimed feeding 4/24/41 : 0.157't : 9.21 : r+ + + : 5/3/41 : 0.218 : 8.55 ,. + + + : 5/17/41 : 0.342 : 10.63 :i- 1- 1- + : 6/7/0. : 0.150 : 8.80 :+ + + 1- : 9/24/41 : - : 5.36 :4 + t i- : 10/1/41 : Start of second period - started feeding 10 grams per day 10/10/41 : 0.290 : 4.076 : 1- 4 e :Continnoi 10/17/41 : 0.250 : 2.502 ~ : + : - 10/24/41 : 0.475 : 5.316 : :- + : « 11/7/a : : ~ : :Discontinued - lost muscle tone 11/8/41 : 0.191* : 5.563 : 4 7 + : 11/12/41 : : : :Started 5 ms. per day 12/2/41 : 0.356 : 7.417 3? + + #QContimed 12/10/41 : 0.310 : 4.18 :1- + + + : I 12/17/41 : 0.366 : 4.532 : + «t 1» : " 1/14/42 : 0.152* : 6.495 :+ + + + : u 1/23/42 : 0.1351"I : 6.158 :1. 4- + + : I 2/5/42 : 0.358 : 9.003 :+ t + + : ' 2/20/42 : 0.257 : 0.931 :Bloody : 9 3/14/42 : 0.290 : : . :Discontimed 3/17/42 = 0.290 : 1.256 3+ + t + 3Sallple by massage 4/7/42 : 0.177 : 4.59 :4 «r r + : 3 3 * Low values due to hemlysis. ”‘ Saen had 90 per cent motility when fresh and 50 per cent at 96 hours. -74- TABLE II. m 01' CHLCROBUT‘ANOL 0N BREEDING EFFICIENCY OF BULL '1645'. 3 3 3 3 Period : beatment : Savices : Conceptions : Services per : : : : Conception : : : : 1st. 11 no. : None : 42 : 11 : 3m : : : : 1st. 5 mo. : 275 gr. : 13 : 5 : 2.60 : : : : 2nd. 3 no. : None : 11 : 1 : 11.00 : : : : aid. 4 no. : 1170 gr. : 7 : 3 : 2.33 3 3 3 3 TABLE 111. mm or FEEDING CELcsosoTANOL To SIRE 72-1 (HCLST‘EIN). 3 3 3 Date :mrbic Acid per 100 ml. :Spern :Chlorobutanol T‘reahent : Bloo d : Semen :Mo til ity : : mg. : g. :(h‘esh) : 3 3 3 3 9/24/41 : 0.323 : 4.081 :e- 4 : 10/1/41 : : : :Star ted feeding 10 5!. daily 10/10/41 : 0.1245:I : 0.646 (1):+ 4 :Contimed 10/17/41 : 0.214 : 2.197 :1. + : I 10/24/41 : 0.504 : - (2): : I 11/8/41 : 0.224 : 4.186 :+ + + : I 11/9/41 : : : :Rednced dosage to 5 g. daily 12/2/4 : 0.276 : 1.935 :4 :5 gr. per day 12/10/41 : 0.208 : - : :I " I I 12/17/41: 0.232 : 1.996 :.+ + :I I I I 1/14/42 : 0.197 : 1.839 :4 z' " " I 1/23/42 : 0.273 : - (3): :l'ed 10 gr. for 9 days then 5 gr. 2/4/42 : 0.484 : 3.75 :- + 1: 1- :5 gr. 2/20/42 : 0.54) : 4.615 :4 4 + + :I I ,_ 3/14/42 : : : :Discontinued feeding 3/16/42 : 0.226 : 2.082 :+ :Nons 4/7/42 : 0.017 : 1.20 :4-4» :" 6/9/42 : 0.147 : 0.920 :l'ew :- 3 3 : ‘ Sligit hmolysis. (1) Semen sample bloody. (2) Not able to get sanpls (he to lack of coordination. (3) Semen thick, bloody mass - only 0.7 co. in sample. -75- 71131.13 n11. misci- or 06106030710101. on BREEDING 27710123307 or 813! 72-3. 3 3 3 Period 3 Treatment 3 Services 3 Conceptions 3 Services per 3 3 3 3 Coantion 3 3 3 3 3 no. Prior 3 None 3 l7 3 3 3 5.66 3 3 3 3 4 no. of treat-- 3 3 3 3 Int 35-10 paper 3 13 3 5 3 2.60 3 3 6.; 7361.1: 1317. mm or mmaau'rm 7620160 10 616: 1601 (HOLSTEIN). 3 3 3®ern 3mlorobntenol Treatment hte decorbic Acid per 100 m1. 3 Blood 3 Semen 3M0 til ity 3 3 m5. 3 .5. 3(rreeh) 3 3 3 3 3 33/1/41 : 0.22 : 6.66 :1. :- + : 33/7/41 : : : 3Started feeding 5 gr. daily 3/22/41 3 0.27 3 8.44 3+ + + 3 33/29/41 : 0.117 : 7.21 :+ .3 : 4/2/0. 3 3 3 3Discontinued feeding 4/6/41 : 0.119 : 6.606 :+ + : 4/7/41 : 0.229 : : 3Stnrted feeding 6 gr. daily 4/9-12/41: : : :10 gr. daily for 3 days 4/12/41 : 0.313 : 10.60 :+ + r + 3Discontimed feeding 3 3 3 TABLE IV. EFFECT OF CHLCBOBUTANOI: ON THE REEDIN} EFFICIENCY OF SIR] 1601. 3 3 3 3 Period 3 Treatment 3 Services 3 Ooncqation 3 Services per 3 3 3 3 Conception 3 3 3 3 1 no. 3 lone 3 6 3 2 3 3.00 3 3 3 3 4 no. 31 m0. gr.3 15 . 11 3 1.36 3 3 3 3 3 3 3 3 MI XVI. -76- 13mm 0? CHLGIOBUTANOL FEEDING T0 SIR] Immmcan". Bite 3 3_A_scorbic Acidper 100 nl. 3Spern3SeInen3Per Cent Motility - 3 3 ”BI 3 Blood 3 Semen 30onc.3 pH 3ft:esh324 hr.348‘nr.3 2 hr. 3 :15. 3 mg. 35/00 3 3 3 3 3 3 3 3 3 3 3 3 3 3/17/42 3 0.241 3 3.3) 3 - 3 - 3 - 3 - 3 - 3 - 3/20/42 3Started feeding 10 grams of chlorobutanol per day 6/26/42 : 0.087 : 4.760 : : : 10: 0 : - : - 4/6/42 : 0.316 : 6.064 : : : 60: 20 : - : - 4/10/42 :rea 16.3 10 a. dose - none 3.6 4/6 an 4/9 - Incoordination 4/17/42 3 0.244 3 1.994 3 3 3 503 10 3 - 3 - 426/42 : 0.166 : 6.906 :1.612: : 70: 20 : - : - 26/42 : 0.222 : 6.615 :.620:6.9o: 70: 60 : 60 : :1) 6/4/42 : 0.297 : 4.629 3 .140: 7.60: 40: o : - : - 5/7/42 : 0.616 : 4.260 :.646: 6.66: 60: 40 : 40 : 16 5 7/42 3Started feeding 40 grains of chlorobutanol per week 6/12/42 : 0.626 : 6.292 :.660:6.76: 76: 10 : 10 : - 6/19/42 : 0.664 : 2.616 :.672: 6.66: 60: 50 : 20 : 10 6/26/42 : 0.664 : 11.66 :.792: 6.76: 90: a) : - : - 5/26/42 3mscontinned feeding chlorobutanol BIZ/a 3 0.097 3 4.132 3.7883 6.603 803 4) 3 a) 3 15 6/9/42 3 0.227 3 4.060 3.3363 6.963 m: 60 3 4) 3 20 7/60/42 : 0.174 : 4.999 : -: -: 66: 76 : 65 : 60 3 3 3 3 3 3 3 3 rm: XVII. PIE cm 01' ABNMMAL SPEMA'I'OZOL IN 13113 SEMEN OF SIRE "SHCB'HCRW ' 3 Date 3 Per Cent Abnormal 3 Comments 3 3 21/42 3 13 3 Dent bodies 5/19/42 : 11 : Bent 66616. and coiled 3.11- 6/9/42 : 9 : I I I I I 7/60/42 3 12 : I I I I I 3 3 -77- TABLE XVIII. mm 0! RIDING 06106060131301. 1'0 THE “VILLS' HILL (JESS!) .ee m M ...5. t e M33M333333W ”00 00 OMMOMO 0 “05m000522 .4 33 33 3 30333 0333333333 3 W M ...m.0 000 50 on” m u m::6:::::-ua-:: :-m1 mum . 6 “16661666: ” m " eeeeeeeeeeeoeeeo eeeeeoeoeeeeeoeeeeee ee 3 4 .5.m5 00 a m::m::::::mwm:u WWW» mmmmmwoo azuwumummu r 6 mm0 1 3333 3333333333 3 m:-m:-,-mmmmwmmm...mmm.»m.mma 62.6.6.6». 1O.” oovsonflosza 00 333333 eoeeeoeeeeeoeoeeeeeo 3 a2 m506476 M33”333WWW1&222M&&$”fl352202Mm MMMMMMMMMM : :::o:: ::::::: m “98 omwmiw4hhu8 3 3333333333 3 3 ”m1 we,w,-n-nnm www.mnm. m37m. m.m6mmm.....mm m M . 6M889m07 0:8 ”$33333333 3333333333 3 6% w n1mao1mm mu 1 n ::::::::..:: u mtm n u no 43 a 33 33 am 33333333h3333333333 3 “Lmn mmwommamwwwmmmmmmz3mflw&“¢WW573w2fl06 .. ozzoo4u11o LL1mu.. one vasonw41wsma mmu,-mu--,,u--,m,-”11mumw-wwmmwszza.aaaz m w. mummwmmwwuumm ammo m:u:::::: : “all e e 1291"..."3' e6$$54fiww awma -,:--mmmmwm,mmmmmmmwwmmmmmmmmmoooomo1ww.1 an :1:--:-:-- » Ammmm4WWMMMMMI4WWe6wmm mmmnm aeowmm.m h ll]h1/tlflhfllflll 384077 [I’llmmyo 44646666 6a:ovnammmflmummwmwwwmmwwwmm a h 4 4 e O t v . .. . . . . l b e . i 1 1 e u e ~ I e . I l 1. . o. 3 . . . . 4 A e o e 4 4 . u. . 1 . , . . . ._ .. . . . . . 4.1.,l it 3.. -73.. TABLE XIX. EFFECT OI' CHLQQBUTANQ: 0N BREEDING MICIENCY 01‘ 'WELS' SIRE. Period Treatment Services Conceptions Services per conception 3 months lone 10 2 5.00 2 months 390 grams 3 2 1.5) TABLE 13:. Pm GET W ABNORMAL SPERMA'IOZOA I}! SHIN Ol' WILLS BULL. at. Per Cat Abnorgl _lomente 4/21/42 14 0611.6 tails 5/5/42 14 . . 5,9,4? 15 s 33 7/16/42 19 n 33 6/24/42 12 u . -79- mu m. mom or monaeumm. mum on 61613 In-620I (6131) 1mm). 3 T 3 3 nte 3 Ascorbic Acid 333cm Semen 3Per Suit Motility of Spa- 3 per 100 ll. 3Conc.3 3_ 3 Bleed 3 mom 3 3 3 3 3 3 3 mg. 3 165. 30/111311 3 pH 3 Vol. 31resh324hr.348 hr. 372 hr. 3 3 3 3 3 3 3 3 3 6/16/4230.1393-3-3-3-3-3-3-3- 6/23/42 3 0.334 3fi1trate destroyed 3N0 notile sperns3considerable sedinent 6/23/42 3 Started feeding 20 grams chlorobutanol by capsule 6/29/42 3 0.324 3 - 3 3 3 uppeared eleepy 6/29/42 to 7/6/42 fed 160 gram of 66161-666366“ by 60 gran doses 111 caplulo for: 7/7/ 42 7/15/ 42 7/15/42 7/22/42 7/26/42 6/4/42 8/10/42 6/17/42 6/17/42 6/24/42 6/61/42 9/4/ 42 9/12/42 9/16/42 9/19/42 9/21/42 9/29/42 10/2/46 3 10/15/423 10/10/42: 10/20/42: 10/27/42: 11/10/42: 11517/42: 12 1/42 3 12 8/42 3 12/12/423 12/17/42: 12/22/42: 1/5/43 3 1/5/43 to 1/16/46 : 1/16/46 1/20/46 1/27/46 1/27/46 2/6/46 2/10/46 2/24/46 3/11/43 3/18/43 4/6/46 ”””~”“””””””N”””” ””"”””..OO“” ova a 5 day period. n'owsy airing this per1od 0.328 3 0.2453 3 6.42 3 1.3 3130 sperns 0.257 3 1.1273 3 7.60 3 2.0 3130 sperms Started feeding 10 gram chlorobu tanol every other day 0.44831.9983 3692313633103 630 30 0.385 31.532: 37.103 2.93 163 6 3 0 3 0 0.332 3 2.0123 3 7.22 3 7.0 316w dead sper- 0.671:6.0003 :- : 6.6:I I I 0.392 3 2.2243 3 7.00 3 3.3 3" " " .Senen brown in color Discontinued feeding chlorobutanol 0.402 3 2.1303 3 6.78 3 2.5 35o sperms 0.255 3 3.4443 3 8.96 3 1.5 37.1: deed sperIs 0.1373-3 3-32.332323230 036133.103 3 - 3 - 3Dead sper- O.226 3 led 25 cc. of shark liver oil containing 25.000 LU. of vitanin A. Increased to 50 cc. shark liver oil every 3 days 0.332 3 2.7923130 motility. Considerable cell debris. 0.267 3 Hasmrbated .. 3 I 0.295 3 2.6823 3 3 1.6 3rew dead spans Started 25 greens chlorobutanol twice a week 0.362 3 4.4503 3 3 2.2 3Iany dead sperms 0.363 3 3.9943 3 7.08 3 0.7 3I I I 0.242 3 3.5833 3 7.12 3 3.3 3Ce11 debris and crystals 0.323 3 3.4953 3 3 0.6 3" " I I 0.254 3 2.7093 3 3 0.4 3" I I I 0.279 3 2.2453 3 6.95 3 9.6 3Seninel fluid Started feeding 50 grams chlorobutanol 2 times per week [as too drowsy to collect sample 0.334 3 [as too drowsy to collect sample 0.2:” 3 0.8703Suina1 fluid 1/12/ 43 - He chlorobutanol - 3 3.5833l'ew club tailed oper- Uterted feeding 5 grans chlorobutanol per (la - 3 - 3 3 3 1.9 3rew dead sper- - 3 3.2103 3 3 3" I I Stepped feeding shark liva‘ oil Stopped feeding chlorotu tanol 0.318 3 2.9903 3 3 0.4 316 sper- 0.364 3 3.2753 3 3 6.4 3" I 0.286 3 2.7563 3 3 1.7 316w deed sperms 0.231 3 1.0313 3 3 3" I I - 3 4.2043 3 3 3Senina1 fluid - no sperms I. C O 1 . . V e e v 1 1 . e s I e . l c 1 1 . 3 53.5335! fluja -80- mu 1111. mm or mmomrmoz. 7136111136 013 61613 ImumrI (6166133). 3 f 3 n3. 3 Ascorbic Acid 3 3 3_per 100 n1. 3 Semen 3Per Cent Ibtility of Spern 3 Blood 3 Semen 3 3 3 3 3 3 3 mg. 3 5. 3pH 3 Vol. 3Presh 3 24 hr. 3 48 hr. 3 72 hr. 3 3 3 3 3 3 3 3 10/10/42 3 - : 2.977 : - : 4.6 : 90 : 60 : 45 : 20 10/22/42 3 0.226 : 4.422: - 3 3.4 : 90 : 66 : 46 : 60 10/29/42 3 - 3 2.632 3Started feeding 10 grams chlorobutanol per day 11/6/42 : - : 2.646 : : 6.6 : 6O 3 46 : 40 3 60 11/10/42 3 0.286 3 10st 3 3 3.2 3 50 3 35 3 a) 3 a) 11/12/42 : - : 6.666 : : 4.1 : 66 3 50 : 4o : 35 11/21/42 Muscular incoordination. lo 611161-666361161 from 11/21 to 11/26 12/1/42 3 0.219 : 2.709 3 7.96: 6.2 : 60 : 15 : 0 3 0 12/16/42 : 0.366 : 2.620 : 7.20: 2.6 3 60 3 15 3 o 3 0 12/16/42 : - 3 2.671 : 7.053 4.2 : 66 : 46 3 o : 60 12/22/42 3 - 3 5.041 3 - 3 4.1 3 75 3 5 3 0 3 0 1/5/43 3 0.341 3 No econ sample taken 1/5/43 3 1/12/43 Muscular incoordination. Io chlorobutanol 1/6/43 3 3 4.390 3 - 3 4.2 3 w 3 35 3 25 3 0 1/11/43 3 3 6.745 3 - 3 3.8 3 85 3 45 3 4) 3 a) g 3 3 3 3 3 3 3 IABLI XXIII. EFFECT OF CHLOROBUMQ: OH BREEDING EFFICIENCY 01' "HEN!” BULL. Period Treatment chices Conceptions Services per Concgption 3 no. previais lone l3 3 4.33 3 1310. following 61 days 7 5 1.0 TABLI 10317. 13771303 or mmowmm 76611610 013 813! IunnI (Jam) n3. 3 Ascorbic Acid 3 3 3 per 100 ml. 3 ger- 3Per Cent Motility of Sper- 3 Blood 3 Semen 3 Vol. 3 3 3 3 3 mg. 3 mg. 3 n1. 3besh 3 24 hr. 3 48 hr. 3 72 hr. 3 3 3 3 3 3 3 1/26/443116 :11636.6I:m: 0: o : 0 1/28/44 3Dsterni1ntion. Started feeding 5 grams chlorobutanol per day 2/16/44 : nade : 6.0 : 46 3 20 : 0 3 0 2/17/44 3 3 3 3.5 3 50 3 20 3 0 3 0 2/25/44 3 3 3 4.5 3 50 3 40 3 25 3 20 6/6/44 : : : 6.6 : 60 : 46 : 10 3 0 6/16/44 3 : : 4.6": 76 3 60 3 40 : 16 4/19/44 : : : 4.03: 10 : 0 3 o 3 0 4/22/44 : : 3 6.4 3 4o : 0 : 0 3 0 4/22/44 3 conti :‘H (0. 3B O 3:6 feedi ng chle robu tanol f 03 .I g .1 ma AW wave. r t ..‘ w.“ .. ...«I .thnw. . s. 3 . . {Hm M! m. MT 0! 'IHE HEREIN} EFFICIENCI 0? EB "LADD' SIRE. Period host-lent Services Conception Servicee per Concep tiong 3 no. previous None 9 l 9.00 3 130. following 425 gm. in 85 days 15 2 7.50 IABLI m1. mwr O! CHLGCIBUTAKOL FEEDING ON RMONCTIW IN DAIRY GOES. MD "A”. 3 3 3 3 3 3 Cow 3 leigit 3 Sires 3 Services 3 usage 3 we 3 Services per 3 3 Used 3 Before 3 per 3 red 3 Conception 3 3 3 Trenton“ 1h: 3 3 3 1.66. 3 No. 3 No. 3 Grass 3 3 N0. 3 3 3 3 T 3 airprise 3 850 3 3 3 6 3 5 3 21 3 1 Joy 3 600 3 3 3 9 3 5 3 21 3 (5) Joy 3 3 3 3 20 3 5 3 (1) Joy 3 3 3 3 5 3 16 3 6 fluid 3 800 3 3 3 6 3 5 3 21 3 1 Jewel 3 850 3 4 3 l4 3 5 3 21 3 (6) Jewel 3 3 3 3 20 3 5 3 (1) Jewel 3 3 3 3 5 3 16 3 7 Enable 3 850 3 3 3 5 3 5 3 21 3 l Marcie. 3 900 3 3 3 8 3 5 3 21 3 (3) larcis 3 3 3 3 20 3 5 3 (1) Heroic 3 3 3 3 5 3 16 3 4 Vere 3 900 3 l 3 5 3 5 3 21 3 l Neidne 3 850 3 2 3 5 3 5 3 21 3 l 733113.333 013-1 3 600 3 6 3 a 3 6 3 21 3 (6) 333113333 01331 3 3 3 3 20 3 6 3 (2) Vslient Girl 3 3 3 3 5 3 16 3 7 3 3 3 3 3 3 -82- TABLI EVILWT 01‘ NCRMAL AND 8300! ‘IBEA'MENT ON MILK PROWCTION,EED 'A'. 3 3 3 3 Cor 3 hpe of 3 Grate in 3 Lbe. of eilk 3 Lbs. of Milk 3 ‘i‘reeteeet 3 usage 3 Before freeuent 3 hiring beehent = 8 3 (jar daw) 3 (per d5?) 3 3 3 3 mrpri se 3 Normal 3 5 3 35 3 35 field 3 " 3 5 3 25 3 20 Bremble 3 " 3 5 3 35 3 32 Vere 3 ' 3 5 3 37 3 35 hide 3 " 3 5 3 35 3 37 Average 3 3 3 §.5 3 31.5 Jubilee 3 amok 3 20 3 21 3 10 Velisnt Girl 3 " 3 23 3 20 3 S Averege 3 3 3 23.5 3 5.6 3 3 3 3 TAIBLI munmncr OF CHLORCRT'I‘ANOL ON REPROIUCTION IN DAIRY CMSJERD '3'. Cor lo. 3 3341333 3 Date of 3 Sgrvice 3 Sire 3 Romerks 3 3 Snrvice 3 3 3 3 3 3 3 3 652 3 1285 39/11/44 3 1 3600-0 3 Brown 3 311/9/44 3 2 3600-! 3 Seis- 3 312/20/44 3 3 3600-0 3 3 33/4/45 3 4 3600-! 3 3 33/25/46 3 5 3600-1 3 3 34/17/46 3 6 3600-] 3 3 34/28/45 3 7 3600-! 3 3 36/11-6/193 3 37.31 6 guns per a: 3 35 31 45 3 S 3600-! Green“ 63 3 117 33 2 3 l 3 3 63363333 3 34/18/45 3 2 3600-6 3 Swiss 3 35/9/45 3 3 3600-! 3 3 35/m/45 3 4 3600-3 3 3 36/60-6/19: 3 313.31 6 gm p63- as: 3 36 1 45 3 6 :wo-Wt 621 3 1377 33 7 45 3 1 3600-2 3 Brown 3 33/29/45 3 2 3600-3 3 Swiss 3 34/17/45 3 3 3 3 3 35/11-6/193 3 3red 5 grams per day 3 35 31 45 3 4 3600-] 3Pr t 334 3‘75—‘674W 2 3 1 3Mseoruc ecid Jersey 3 32/7/45 3 2 3330-0, 3 3 32/27/45 3 3 33304 3 3 :4/11/46 3 4 3310-0, 3 3 35/12—6/193 3 31-631 6 guns per dey 3 36L350L46 3 6 3600-31 3193-3133363 ML] 117111 - Continued 3 3 3600-! 3Pregnent 3 3 60'. lo. 3 '4‘“ 3 lite of 3 Service 3 Sire 3Renerke 3 3 Service 3 3 3 3 3 3 3 3 .3—6 : 766 :1/22/46 3 1 3350-0 3 Jersq 3 32/10/46 3 2 3650-4 3 3 :2/14 46 3 6 3330-0 3 3 33/5 45 3 4 3310-0, 3 3 33/25/45 3 5 3330-0, 3 3 36/12-6/193 3 3364 6 grens per 4c 3 36 8 45 3 6 3M 3Pr met 53-! 3 11 3 2 3 3 3 3 35/2/45 3 2 3330-0, 3 3 35/23/45 3 5 3330—0, 3 3 36/21/45 3 4 3330-4 3 3 35/26-6/193 3 xl‘ed 5 grees per as: 3 38 l 45 3 5 3Wt 7:2 3 3 3 l 3 3 ieTescorFic eoid Jersq 3 32/19/45 3 2 3610-0 3 i 83/26/45 3 6 3600-0, 3 3 34/19/46 3 4 3600-0, 3 3 35/9/45 3 5 3&0-0, 3 3 36/12-6/193 3 3J'ed 6 grene per day 3 35 3O 45 3 6 3m 3Pre at 3:3 3 550 3251/& 3 I 35% 3EIed escorbic ecid .3ch 3 32/22/45 3 2 3330-0, 3 3 33/15/45 3 3 3330-0, 3 3 34/19/46 3 4 3610-0 3 3 35/12-6/193 3 Wed 5 gram: per day 3 36 15 45 3 5 3Wt If 3 1r/77L6 36 45 3 1 3o 3 Ayrshire 3 34/17/45 3 2 3430.! 8 3 35/11-6/193 3 3red 5 grens per dew 3 35/39/45' 3 3 3430-! 3 3 36 9 45 3 4 3430-! 3Pr t 3'54— 3 IE§33/£735 3 I 350:]: 3 Brown 633134 3 34/17/46 3 2 3600-13 3 3 35/9/45 3 3 3600-] 3 3 35/11-6/193 3 3red 5 gram per m 3 35/30/45 3 4 36m-B 3 3 36 26 45 3 5 3600-! 3Pre t W 3 1% 31271513 3 1 32333338. Bridge3 Brown Swiss 3 31/6/45 3 2 3Denonetrator 3 3 31/27/46 3 6 31363363336336: 3 3 33/7/45 3 4 3 " 3 3 33/29/45 3 5 3600-1 3 3 34/18/45 3 6 3600-: 3 3 35/11-6/193 3 3red 6 grene per 313: 3 35/23/45 3 7 3600-! 3 3 3 8 3 3 36/22/45 3 3 3 -84- 661333133 3 1067 34/14/44 3 1050 :6/16/44 3 1045 37 14/44 661333133 3 1611 310/16/44 3 " 3. 3Pml 3Sterted 3 gm. chlorobntenol deily 392144 3 1667 311/4/44 3 1605 31/12/46 3 1606 32/24/45 1697 33/19/ 46 1.461.: 1111. m 03' (HLOBOHITANOI. 033 61313063011013 113 311131 0076.6n6 '0'. 3 3 3 3 3 00' lo. 3Ieid1t3 Dete of 3S.rvice3 Sire 3 Remarks 3 3Mice 3 3 3 3 3 3 3 3 1-27 3 966 31/24/44 3 1 321-111: 3 631334133 3 962 :2/16/44 3 2 3 33 3 3 972 36/6/44 3 6 3 3 3Sterted 6 gl. 63313336333333.3361 31.11: 3 964 36 24/44 3 4 3 '8 :Pr. 3 6/6744 32E 3 1166 :6 6/44 3 T :m—Er d3 ' 631334133 3 1060 37/16/44 3: 2 333361 3 31099 38/5/44 3 6 3 33 3 3 1064 38/25/44 3 4 3 3 363333-3631 4 p.0h10rohtm1 9/60/44 31038 39/15/44 3 5 3 " 3 3 1123 310 28 44 3 5 deirfield3PreQnt A- ~ 3 423 3 3Pril y 3 116133316 3 964 36/6/44 3 2 3 33 36363-33431 6 gl. 63316366636661 per 633: 3 976 36/24/44 3 6 39313-316131: 6f6/44 3 960 3 16 44 3 4 3 3 393-6 3333-1663-3331 11/15/44 37 3 32 3 3 r 3 1361336133 3 1066 34/4/44 3 2 3 :3 393333-3631 6 gm. 36133663336661 daily 3 1060 34/26/44 3 6 3Pen1 3 5/10/44 3110036844 3 4 3 " 315-313;“ 3 596 312 31 43 3 l 3Prilly 3 86133.13 3 966 33/14/44 3 2 3rdl‘f101d3 31017 34/14/44 3 3 3 ' 3 A 3 1010 36/4/44 3 4 313-31 36341-3631 6 533. 33331633613333.3361 daily 3 1025 35 20144 3 6 31.131141131139333 6/10/44 33 3 31 HIE 3 l 3 orne 3 Jersey 3 706 34/10/44 3 2 31.333131 3 3 700 35/3/44 3 3 31'110rne 3 3 760 :6/27/44 3 4 3 - 3Sterted 6 a. 33316366634161 daily 3 760 37 61/44 3 6 36.31% 3131-23931 6110/44 2 3 10 33 2 44 3 l 3 ir el 3 3 2 3 3 3 4 3 l 3 2 3 3 3 4 3 5 3 6 3 7 , _ 1696 :4/12/46 _3_1'41rr141313r_3-_open3 6116144 31'eirfield3 3Pe11 3 3 " 31‘sd 4 an. chlorobutanol daily 3 ' 311/8/44 to 1/25/45 3 " 3 3 " 3Stertsd 8 9|. chlorobutanol deily 3 " 3Pregnent 3/33/45 TABLI m. EFFECT 03* CHLGROBUTANOL 0N REPBOIIICTION IN DAIRY COWS,HED '1’. V 3 3 3 3 - 3 00' zloightflhnber of 3800-7100: 31136" porxmyomunber of 3 xsiroo Una: before 3 my mod 3591-vicol 3 3 3'rroatnont3 3 330 Conceive 3 3 3 3 Gran: 3 3 3 3 3 3 3 3 $410 I 3 1350 3 2 3 4 3 5 3 35 3 1 330335611 3 1000 3 1 3 4 3 5 3 24 3 1 Admiral'o Clara 3 1100 3 2 3 1 3 5 3 20 3 (6) " I 3 3 3 5 3 3 31 3 (2) I " 3 3 3 3 5 3 24 3 6 Admit-61's Mary 310003 2 3 4 3 5 3453 2 3 3 3 WW 0? limes Bred without concoiviniaTtor flea-ing for days shown TABLE mu. m 01' mammal. MD!“ G FLASH ABCCEBIC ACID AND LIBIDO 3 3 3 3 Sir. 3 Into 31’1”“ hoorbic330rvico Tino3ch10r0‘mtanol has. 3 3 Acid. 3 3 3 3115 per cont 3 3 3 3 3 3 Patty 32/28/0. 3 .22 3 10-15 33133. 3133333. 36/7/41 3 .28 3 5 I 36 3133. 43433431 2/28/41 36/14/41 3 .41 3 1 I 3 I I 33/22/41 3 .51 3 5 000. 3 " 33 29 41 3 .27 3 5 " ' Pheiffor Boy 3 2 12 41 3 3 I 53. 330310 312/16/41 3 .668 3 310 gm. started 12/12/41 312/60/41 3 .660 3 15 .133. 3 I .3 3113/42 3 3 5 ' 3130310 MIC CHIGAN STATE UNIVERSITY LIBRARIES I III II III III I III III III I II