Estimation of inbreeding and effective population size of full-blood Wagyu cattle registered with the American Wagyu Cattle Association.

The objective of this research was to examine the population structure of full-blood (100%) Wagyu cattle registered in the United States with the American Wagyu Association, with the aim of estimating and comparing the levels of inbreeding from both pedigree and genotypic data. A total of 4132 full-blood Wagyu cattle pedigrees were assessed and used to compute the inbreeding coefficients (FIT and FST ) and the effective population size (Ne ) from pedigree data for the period 1994 to 2011. In addition to pedigree analysis, 47 full-blood Wagyu cattle representing eight prominent sire lines in the American Wagyu cattle population were genotyped using the Illumina BovineSNP50 BeadChip. Genotypic data were then used to estimate genomic inbreeding coefficients (FROH ) by calculating runs of homozygosity. The mean inbreeding coefficient based on the pedigree data was estimated at 4.80%. The effective population size averaged 17 between the years 1994 and 2011 with an increase of 42.9 in 2000 and a drop of 1.8 in 2011. Examination of the runs of homozygosity revealed that the 47 Wagyu cattle from the eight prominent sire lines had a mean genomic inbreeding coefficient (FROH ) estimated at 9.08% compared to a mean inbreeding coefficient based on pedigree data of 4.8%. These data suggest that the mean genotype inbreeding coefficient of full-blood Wagyu cattle exceeds the inbreeding coefficient identified by pedigree. Inbreeding has increased slowly at a rate of 0.03% per year over the past 17 years. Wagyu breeders should continue to utilize many sires from divergent lines and consider outcrossing to other breeds to enhance genetic diversity and minimize the adverse effects of inbreeding in Wagyu.

Effects of immunization against luteinizing hormone-releasing hormone and treatment with trenbolone acetate on reproductive function of beef bulls and steers.

The objectives of this study were 1) to evaluate the ability of trenbolone acetate (TBA) administered in tandem with LHRH immunization to suppress reproductive function in bulls and 2) to examine the effects of LHRH and androgen (TBA) signaling on pituitary gland function. Forty-four Angus × Hereford crossbred calves (BW=225 ± 2 kg; age=187 ± 6 d) received castration, LHRH immunization, or TBA administration in a 2 × 2 × 2 factorial design. Treatment groups receiving LHRH immunization contained 6 animals, whereas other treatment groups contained 5 animals. Animals immunized against LHRH received a primary injection and 2 booster injections of ovalbumin-LHRH-7 fusion protein on d 0, 42, and 196, respectively. Animals treated with TBA were implanted on d 224. Serum LHRH antibodies increased (P<0.05) after each booster for immunized animals, but were negligible in nonimmunized animals throughout the experiment. Serum testosterone concentration (P<0.001) and scrotal circumference (P<0.05) were depressed in LHRH-immunized bulls compared with nonimmunized bulls by d 84 and 168 of the experiment, respectively. Treatment with TBA tended (P=0.08) to decrease serum testosterone concentrations of nonimmunized bulls. Weights of testes at slaughter were decreased (P<0.001) for LHRH-immunized (232 ± 41 g) compared with nonimmunized (752 ± 45 g) bulls, but did not differ (P=0.80) between TBA-implanted (500 ± 49 g) and nonimplanted bulls (484 ± 36 g). Both LHRH immunization and castration decreased pituitary gland stores of LH and FSH (P<0. 001). There was no effect (P>0.10) of TBA on pituitary gland FSH content and only a tendency (P=0.09) to increase pituitary gland LH content. Immunization against LHRH decreased expression of LH ß-subunit and common α-subunit genes (P<0.001). Castration increased expression of LH ß-subunit and common α-subunit genes (P=0.02). Treatment with TBA further suppressed (P=0.04) α-subunit mRNA expression in LHRH-immunized steers. In summary, LHRH immunization decreased synthesis and storage of LH and decreased storage, but not synthesis of FSH in bulls. The increased synthesis of LH and FSH in nonimmunized, but not LHRH-immunized steers suggests that castration removes the negative feedback on gonadotropin synthesis but that LHRH is still needed for release of these hormones. Androgen replacement with TBA did not restore the negative feedback control of gonadotropin synthesis.

GnRH or eCG treatment fails to restore reproductive function in GnRH immunized ewes.

This study was designed to evaluate the potential of using eCG or GnRH in restoring reproductive functions in GnRH immunized ewes. Thirty-three multiparous Kivircik ewes were randomly assigned into either control group (n=11) or immunization group (n=22). Ewes were immunized against GnRH by injecting with a cocktail of ovalbumin-LHRH-7 (ovalbumin-GnRH-7) and thioredoxin-LHRH-7 (thioredoxin-GnRH-7) fusion proteins generated by recombinant DNA technology in April. 500 IU eCG or 0.008 mg GnRH analogue was used to induce ovulations. Serum GnRH antibodies were present in animals of the immunized group beginning the second week after the first immunization and maintained throughout the study (14 months). Immunization caused anestrus in immunized ewes. eCG or GnRH analogue administration given after 14 days progestagen (20 mg fluorogestone acetate, FGA) treatment during breeding season (mid July) did not induce ovulation in these ewes. Two more attempts with single or multiple eCG injections failed to induce ovulation in this group as well. It appears that the gonadotropin stimulation was not of adequate time since neither eCG nor GnRH administration was able to restore reproductive function in immunized animals. The immunization effect lasted more than a year. These results suggest that GnRH immunization exerts its effect via the hypothalamo-pituitary axis and that more than such stimulation is required to overcome the reproductive suppression.

LHRH fusion protein immunization alters testicular development, ultrasonographic and histological appearance of ram testis.

This study was designed to evaluate the effectiveness of a luteinizing hormone releasing hormone (LHRH) fusion protein immunization on reproductive traits in ram lambs including the changes in histology and ultrasonographic appearance of testis. Thirty native ram lambs at 19 weeks of age were divided into control (C, n = 10), immunization (I, n = 10) and castration (E, n = 10) groups. Animals in immunization group were immunized against LHRH using ovalbumin-LHRH-7 (OL) protein generated by recombinant DNA technology as a primary and a booster injection at 19 and 23 weeks of age respectively. Animals were bled via jugular venepuncture at 2-week intervals to determine LHRH antibody and testosterone concentrations. Bi-weekly ultrasonographic examination of the testes was performed to determine the changes in ultrasonographic appearance as the age increased. Biopsied testicular tissues taken at 19, 29 and 41 weeks age were also evaluated. At 41 weeks of age, animals were slaughtered. Semen and epididymis were evaluated for the presence of sperm cells. Testicular development and sperm production were suppressed in the immunized animals. Semineferous tubule diameters decreased, basal membrane of the tubule was thickened and hyalinized in immunized ram lambs. While testes of control animals gained their normal ultrasonographic appearance as the age increased, immunized animals had uniform hypoechogenic testicular structure as observed at 19 weeks of age until slaughter. Simultaneous histological and ultrasonographic evaluations indicated that the changes in testicular histology could partly be monitored via ultrasonographic imaging. Nevertheless, it is difficult to claim that ultrasonographic image reflects the exact changes in such instances. In conclusion, these results indicate that recombinant OL fusion protein is effective in immunocastration in ram lambs and has a potential to be used as an alternative to physical castration. Further research studies should be conducted to help assess reproductive status of testes from ultrasound images.

Changes in testicular development, ultrasonographic and histological appearance of the testis in buck kids immunized against LHRH using recombinant LHRH fusion protein.

This study was designed to evaluate the effectiveness of recombinant Ovalbumin-LHRL (OL) immunization on changes in testicular size, histological appearance and testosterone production in buck kids. Thirty native buck kids at 18 weeks of age were divided into three groups, control (n = 10), immunization (n = 10) and castration (n = 10) groups. Immunized animals received OL protein generated by recombinant DNA technology. Ultrasonographic and histological examinations of the testes were performed. Animals were slaughtered at 44 weeks of age. Semen and epididymides were evaluated for the presence of sperm cells. Immunized animals generated anti-LHRH antibodies. Testosterone production, testicular and accessory glands development and sperm production were suppressed in the immunized animals (p < 0.01). Semineferous tubule diameters decreased (p < 0.01), basal membrane of the tubule was thickened and hyalinized in immunized kids. Immunization affected ultrasonographic appearance of the testes drastically. While testes of control animals gained their normal ultrasonographic appearance as the age increased, immunized animals had uniform hypoechogenic testicular structure as observed at 18 weeks of age until slaughter. Simultaneous histological and ultrasonographic evaluations indicated that the changes in testicular histology could partly be monitored via ultrasonographic imaging; nevertheless, it is difficult to claim that ultrasonographic image reflects the exact changes in such instances. In conclusion, these results indicate that recombinant OL fusion protein is effective in immunocastration in buck kids and has a potential to be used as an alternative to physical castration. Further researches should be conducted to help assessing reproductive status of testes from ultrasound images.

A researcher's perceptions of United States Department of Agriculture funding in animal reproduction.

Dedicated funding for animal reproduction did not start until 1985 and was available primarily in the reproductive efficiency and physiology areas of the Animal Science Program. Funding for individual grants and duration of funding were similar between the National Institutes of Health and the USDA, typically in the range of 3 yr, with total direct costs of $150,000. The names of these programs have changed over time; the National Research Initiative Competitive Grants Program started in 1991 with a program in animal reproduction. The USDA did not change the award size for individual grants until 2001, when it gradually increased through 2003. The USDA then markedly increased individual grants in 2004 to a funding level of $300,000 to $500,000 over 3 to 4 yr. This has been beneficial for the funded scientist but discouraging to the applicants with high-ranking nonfunded grants. The number of grants funded per year is approaching a low critical number, with an average of only 10 new grants funded per year. At the present funding level it will be difficult for even the best scientist to sustain a research career based only on USDA funding.

Use of recombinant gonadotropin-releasing hormone antigens for immunosterilization of beef heifers.

The objectives of this study were to evaluate the effects of immunization against recombinant GnRH fusion proteins and growth promotants on onset of puberty, feedlot performance, and carcass characteristics of beef heifers. Heifers were immunized against an ovalbumin fusion protein containing 7 GnRH peptides (oGnRH, n = 12), a thioredoxin fusion protein containing 7 GnRH peptides (tGnRH, n = 12), a combination of oGnRH plus tGnRH (otGnRH, n = 12), or a combination of ovalbumin and thioredoxin (control, n = 11). Each heifer received a primary immunization containing 1 mg of protein in 1 mL of adjuvant injected into the mammary gland at wk 0 (mean age = 38 wk) and booster immunizations at wk 6 and 12. Six heifers within each treatment received Synovex H implants at wk -2. Weekly blood samples were collected from wk -2 to 26 for determination of serum progesterone concentrations and GnRH antibody titers. In GnRH-immunized heifers, GnRH antibody titers increased after the first booster injection, peaked after the second booster injection, and remained elevated through the end of the study (P < 0.01). Heifers immunized against oGnRH achieved greater (P < 0.05) GnRH antibody titers than tGnRH heifers but did not differ (P = 0.20) from otGnRH heifers. During the 26-wk study, ovulation was prevented (P < 0.05) in 10 out of 12, 12 out of 12, 11 out of 12, and 0 out of 11 tGnRH, oGnRH, otGnRH, and control heifers, respectively. At slaughter, uterine weights were lighter (P < 0.01) for GnRH-immunized heifers than control heifers. Synovex H-implanted heifers had greater (P < 0.05) ADG from wk -2 to 26, greater LM area, and lesser percentages of KPH, yield grade, and quality grade than nonimplanted heifers, regardless of the immunization treatment. Immunization against GnRH fusion proteins resulted in production of antibodies against GnRH that prevented ovulation in 92% of the heifers without affecting feedlot or carcass performance. Implanting heifers with Synovex H improved ADG, LM area, and yield grade. Improvements in delivery of the oGnRH vaccine may provide a feasible alternative to surgical spaying of heifers.

Reproductive characteristics of grass-fed, luteinizing hormone-releasing hormone-immunocastrated Bos indicus bulls.

Two field trials were conducted in Brazil to evaluate LHRH immunocastration of Bos indicus bulls (d 0 = 2 yr of age). In Study I, 72 bulls were assigned randomly to one of three treatment groups: LHRH0-immunized, castrated, and intact. Immunized animals (n = 25) received a primary and two booster injections of ovalbumin-LHRH-7 and thioredoxin-LHRH-7 fusion proteins on d 0, 141, and 287. Twenty-three bulls were surgically castrated on d 141, and 24 served as intact controls. All animals were slaughtered on d 385, at approximately 3 yr of age. In Study II, 216 bulls were assigned randomly to the same three treatments as in Study I; however, because of a drought in the area, bulls were kept on pasture an additional year, and a fourth treatment was added, in which one-half the LHRH-immunized bulls received an additional booster on d 639 (fourth immunization). All animals in Study II were slaughtered on d 741 (4 yr of age). Luteinizing hormone-releasing hormone antibodies increased following each immunization for immunized bulls, but they were not detectable in castrate or intact animals in either study. Consequently, scrotal circumference was suppressed in immunized bulls compared with intact controls in both studies. By d 287, serum concentrations of testosterone in LHRH-immunized bulls were decreased compared with intact controls (P < 0.01). In both studies, testes and epididymal weights at slaughter were greater (P < 0.01) for intact (500 +/- 17 and 60 +/- 2 g, respectively) than for immunized bulls (173 +/- 22 and 26 +/- 2 g, respectively) and fourth immunization bulls (78 +/- 23 and 20 +/- 2 g, respectively; Study II). At the end of each study, BW was greater (P < 0.01) for intact bulls than for castrated and LHRH-immunized animals. In these two studies, the efficacy of the LHRH fusion proteins to induce an effect similar to that of surgical castration was considered 92 and 93%, respectively. These data support the concept that immunocastration of bulls at 2 yr of age was successful and that it has practical application as a tool for producing grass-fattened bulls in Brazil.

Changes in spermatogenesis and endocrine function in the ram testis due to irradiation and active immunization against luteinizing hormone-releasing hormone.

Spermatogonial stem cell transplantation is a technique that has potential in livestock to enhance genetic gain and generate transgenic offspring through the male germ line. A means for depletion of endogenous germ cells in a recipient's seminiferous tubules is necessary for this technology to be applied. The objectives of this study were to evaluate several methods for depletion of endogenous germ cells in the testes of adult rams and to evaluate ultrasound-guided injections into the rete testes as a means for infusing a suspension into the seminiferous tubules. Sixteen adult rams were randomly divided into 4 treatment groups (n = 4 per group). Treatments consisted of active immunization against LHRH (IMM), localized testicular irradiation (IR), LHRH immunization + irradiation (IMM+IR), and untreated control. Serial bleedings were conducted pretreatment and monthly after treatment for 4 mo, at which time all rams were castrated. Both IMM and IMM+IR rams received exogenous gonadotropin in the form of Perganol weekly for 8 wk before castration to bypass the immunization. All rams also received an ultrasound-guided injection of PBS containing 0.4% trypan blue into the rete testis of one testicle before castration. Rams receiving IMM and IMM+IR treatments had higher (P < 0.05) average percentages of seminiferous tubule cross sections with depleted germ cells compared with controls. Serum testosterone was decreased (P < 0.05) in IMM and IMM+IR rams 1 mo after treatment and throughout the remainder of the study compared with controls and IR rams, which were not different from each other. Serum inhibin concentration was unchanged in all rams following treatment indicating that Sertoli cell function was unaltered. A greater (P < 0.05) average percentage of the total testicular area could be filled with the trypan blue solution by rete testis injection in IMM and IMM+IR rams. These data demonstrate the depletion of endogenous germ cells in adult ram testes without alteration of Sertoli cell viability and function that have potential as methods for preparing recipient animals for germ cell transplantation.

Luteinizing hormone-releasing hormone fusion protein vaccines block estrous cycle activity in beef heifers.

Two LHRH fusion proteins, thioredoxin and ovalbumin, each containing seven LHRH inserts were tested for their ability to inhibit estrous cycle activity. The objective was to evaluate immune and biological responses from alternating the two fusion proteins in an immunization schedule. One hundred ten heifers were divided equally into 11 groups. Two control groups consisted of either spayed or intact, untreated heifers. Heifers in the other nine groups were immunized on wk 0, 4, and 9. Treatments were immunizations of the same protein throughout or alternating the proteins in different booster sequences. Blood was collected weekly for 22 wk, and serum was assayed for concentrations of progesterone and titers of anti-LHRH. At slaughter, reproductive tracts were removed from each heifer and weighed. Heifers with >or=1 ng/mL of progesterone were considered to have a functional corpus luteum and thus to have estrous cycle activity. All LHRH-immunized groups of heifers had a smaller (P < 0.05) proportion of heifers showing estrous cycle activity after 6 wk than the intact, untreated control group. There was no difference in number of heifers cycling between the immunized groups and the spayed heifers during wk 9 to 22. Anti-LHRH did not differ among immunized groups during wk 1 to 9. Starting at wk 10 and continuing through the conclusion of the study, there was an overall difference among treatment groups for anti-LHRH (P < 0.05). Uterine weights differed among treatments (P < 0.05), with intact control animals having heavier uteri than all other groups (P < 0.05). Uterine weights were negatively correlated with maximum LHRH antibody binding (r = -0.44). In summary, the LHRH fusion proteins were as effective as surgical spaying in suppression of estrous cycle activity, but alternating the two proteins in an immunization schedule did not enhance the immunological or biological effectiveness of the vaccine.

Effects of biological type and dietary fat treatment on factors associated with tenderness: I. Measurements on beef longissimus muscle.

The objective was to evaluate chemical, mechanical, and sensory attributes associated with tenderness in divergent cattle breeds--Wagyu (W; n = 12), Limousin (L; n = 12) and F1-cross (WxL; n = 12)--fed two dietary treatments (0 or 6% sunflower oil (DM basis)). A randomized complete block repeated measures design in a 3 x 2 factorial arrangement of treatments was used, and effects of breed, diet, block, and associated interactions were tested. Cattle were fed barley-based diets for an average of 259 d. Twenty-four hours postmortem (PM), steaks from the longissimus muscle (LM) were sliced, vacuum-packaged, aged (1, 3, 7, 14, 28, and 56 d PM) at 2 degrees C, and frozen (-40 degrees C) until analyzed. Wagyu steaks had lower (P < 0.05) Warner-Bratzler shear force (WBSF) values than L steaks across all aging times. At 1 d PM, W steaks required slightly more (P > 0.05) force to shear than WxL or L (0.30 and 0.11 kg, respectively); however, by d 14 PM, W steaks required 0.77 kg less (P < 0.05) force to shear than L. Wagyu steaks received higher (P < 0.05) sensory panel sustained tenderness scores at d 14 PM than L. The pH decline was slower (P < 0.05), and temperature decline more (P < 0.05) rapid, in W carcasses than L or WxL carcasses. Breed and diet did not affect (P > 0.10) free calcium levels (FCL) over time (0, 1, 3, 7, and 14 d PM), 0-h calpastatin activity (CA), d-1 percent collagen (OH-PRO), or d-1 collagen cross-linking (HP). Western blot analysis for the presence of the troponin-T (TNT) 30-kDa fragment, conducted only on samples from steers fed the 0% sunflower oil diet, demonstrated more proteolysis by d 3 PM in L than W or WxL. Overall, breed differences in mechanical and sensory measures of tenderness were not explained by FCL, CA, OH-Pro, and HP. Even though the initial appearance of the TNT 30-kDa fragment was greater in L, linear slopes for appearance of TNT degradation product across aging time were greater for W and WxL (P < 0.01 and P = 0.056, respectively) than for L, suggesting that tenderness differences due to breed may have been facilitated by more-rapid proteolytic degradation over time.

Effects of biological type and dietary fat treatment on factors associated with tenderness: II. Measurements on beef semitendinosus muscle.

The objective of this study was to evaluate attributes in semitendinosus muscle (ST) associated with tenderness in divergent breeds--Wagyu (W; n = 12), Limousin (L; n = 12), and Wagyu x Limousin cross cattle (WxL; n = 12)--fed two dietary treatments (0 or 6% sunflower oil, DM basis). A randomized complete block repeated measures design with a 3 x 2 factorial arrangement of treatments was used to measure effects of breed, diet, block, and associated interactions. Cattle were fed barley-based diets for an average of 259 d. Temperature and pH were measured at 0, 1, 3, 6, 12, and 24 h postmortem (PM). Steaks from the ST were removed 24 h postmortem, vacuum-packaged, aged (1, 3, 7, 14, 28, and 56 d postmortem) at 2 degrees C, and frozen (-40 degrees C) until analyzed. Dietary treatment did not (P > 0.10) affect Warner-Bratzler shear force (WBSF), collagen amount (OH-PRO) or cross-linking (HP), temperature, or pH. Steaks from WxL aged 14 d postmortem had lower (P < 0.05) WBSF values than L (W were intermediate). Cooking time was longer (P < 0.01) in W and WxL than in L; however, breed did not affect (P > 0.10) cooking loss. Cooking time was not influenced by diet, but steaks from cattle fed 6% sunflower oil had lower (P < 0.05) cooking losses. Temperature decreased more (P < 0.05) rapidly, and pH more slowly (P < 0.05), in W and WxL than L in the first 24 h postmortem. Limousin steaks were lighter (higher L*) and more yellow (higher b*) in color than steaks from W and WxL (P < 0.05). The control diet (no oil added) resulted in steaks that were lighter (P < 0.05) than the treatment diet (6% added sunflower oil). Neither breed nor diet affected (P > 0.10) OH-PRO or HP concentration. The results of this study indicate that biological type differences may not be as great in the ST as in longissimus muscle; thus, to increase tenderness in ST, emphasis may need to be placed on processing and cooking techniques rather than genetic selection.

Growth and carcass characteristics of pasture fed LHRH immunocastrated, castrated and intact Bos indicus bulls.

The effectiveness of a luteinizing hormone-releasing hormone (LHRH) fusion protein vaccine or surgical castration, at two years of age, on growth and carcass characteristics of Bos indicus bulls was evaluated. Seventy Nelore-cross bulls were divided into three groups: (1) immunized, (2) castrated and (3) intact control. At slaughter (three years of age), intact bulls had higher body weights, ADG, carcass weights, and muscle percentage compared to immunized and surgically castrated animals. Both castrated and immunized animals had greater marbling and percent carcass fat than the intact bulls. Average tenderness scores were inferior for intact bulls compared to immunized and castrated animals, but these differences were not significant (P>0.05). Juiciness, flavor, thawing, nor cooking losses differed significantly among the three groups. Immunocastration was effective in producing carcass traits similar to that of surgical castration. Therefore, immunization with LHRH fusion proteins appears to have practical utility in the management and castration of grazing bulls.

Growth, carcass characteristics, muscle conjugated linoleic acid (CLA) content, and response to intravenous glucose challenge in high percentage Wagyu, Wagyu x Limousin, and Limousin steers fed sunflower oil-containing diet.

The effect of breed and diet on insulin response to glucose challenge and its relation to intramuscular fat deposition was determined in 36 steers with 12 each of greater than 87% Wagyu (referred to as Wagyu), Wagyu x Limousin, and Limousin breeds. Weaned steers were blocked by weight into heavy, medium, and light calves and placed in six pens with two pens per weight type and with two steers of each breed per pen. Three pens with steers from each weightclass were fed backgrounding and finishing diets for 259 d, while the other three pens were fed the same diets where 6% of the barley grain was replaced with sunflower oil. Prior to initiation of the finishing phase of the study the intravenous glucose tolerance test (VGTIT) was conducted in all steers. Once steers were judged as carrying adequate 12th-rib fat, based on weight and days on feed, they were harvested and graded and samples of the longissimus muscle were procured for determination of fat content and fatty acid composition. Dietary oil improved (P = 0.011; 0.06) ADG and feed conversion efficiency of steers during the latter part of backgrounding and only ADG during early part ofthe finishing period. Generally percent kidney, pelvic, and heart fat was the only adiposity assessment increased (P = 0.003) by dietary oil. The IVGTT results indicated that insulin response to intravenous glucose was lower in Limousin steers than in Wagyu steers. Dietary oil decreased (P = 0.052) fasting plasma insulin concentration in Wagyu steers compared with Limousin steers. The correlation coefficients among the IVGTT measures and intramuscular fat content or marbling score were less than 0.4, and only a negative trend existed between fasting insulin and USDA marbling scores. However, the carcasses of the Wagyu steers graded US Choice, and 66% of the Wagyu carcasses graded US Prime, which were substantially better than the quality grades obtained for the carcasses from the other breed types. Dietary oil did not affect muscle fat content but increased (P = 0.01) conjugated linoleic acid (CLA) concentrations by 339%. Results indicated that IVGTT measures were not appropriate indices of marbling potential in cattle and that dietary oil can enhance CLA content of beef.

Endocrine, growth, and carcass characteristics of bulls immunized against luteinizing hormone-releasing hormone fusion proteins.

This study evaluated the effectiveness of a LHRH fusion protein vaccine on endocrine changes, feedlot performance, and carcass quality of bulls compared with steers and hormone-implanted steers. Crossbred bulls (n = 30; mean weight, 179 +/- 4 kg; mean age, 130 +/- 2 d) were randomly assigned to three treatment groups: 1) castrated (castrated; n = 10); 2) castrated-implanted with trenbolone acetate (implanted; n = 10); and 3) immunized against a cocktail of recombinant fusion proteins, ovalbumin-LHRH-7 and thioredoxin-LHRH-7 (immunized bulls; n = 10). Blood was collected every 2 wk to evaluate antibody and hormone concentrations. Serum LHRH antibodies (P < 0.001) were detected in animals of the immunized group, which had reduced serum LH concentrations (P < 0.001) compared with the castrated groups and serum FSH concentrations, which did not decrease but were significantly different when compared with castrated and implanted animals. Serum testosterone concentrations in the immunized bulls were not different from the two castrated groups (P > 0.05) by d 60 after primary immunization. Initial mean scrotal circumference of the immunized bulls was 18.0 +/- 0.6 cm on d 0 and increased to 22.6 +/- 1.3 cm by d 310. No differences (P > 0.05) in ADG were observed among treatment groups. Immunized animals had an intermediate BW gain (P > 0.05) when compared with the castrates, whereas the castrated groups differed (P < 0.05) from each other. Carcass characteristics were similar (P < 0.05) among the three groups. Vaccinating bulls against a LHRH fusion protein cocktail suppressed LH and testosterone, which led to reduced testicular development and no bullock carcasses. Growth and carcass characteristics of the immunized animals were similar to the steers.

Transplantation of bovine germinal cells into mouse testes.

To develop techniques for spermatogonial transplantation in bulls, it is essential to have an effective bioassay procedure to evaluate the transplantation efficiency of spermatogonial stem cell collection, purification, and culture techniques. The objective of the present study was to develop a mouse bioassay model to evaluate transplantation efficiency of fresh and cultured bovine germ cells. Bull calves of four ages (1, 2, 3, and 4 mo) were used as a source of donor testes cells. Two calves were used for each age point, one calf was experimentally made cryptorchidistic at 1 wk of age and the other left normal. A STO (mouse fibroblast) feeder cell line was used to culture bovine testes cells for 2 wk preceding transfer into recipient testes. Immunodeficient nude mice (nu/nu) in which endogenous spermatogenesis had been abolished by busulfan treatment served as recipient animals for transplantation. Donor bovine germ cells were microinjected into mouse seminiferous tubules. Mouse testes were analyzed 2 wk after transplant with the use of a bovine-specific antibody and whole-mount immunohistochemistry for the presence of bovine donor germ cells. Bovine testis cells were present in all recipient mouse testes analyzed. Fresh bovine testes cells were observed as colonies of round cells within mouse seminiferous tubules, indicating spermatogonial expansion and colonization; however, cultured bovine testes cells appeared as fibrous tissue and not as spermatogenic colonies. The average number of colonies resulting from donor cryptorchid testes was not different (P > 0.05) from noncryptorchid, 56+/-4 and 78+/-7, respectively. Fresh donor cells from calves older than 1 mo gave rise to a greater average number of colonies within recipient testes (P <0.05) (1 mo, 33+/-4; 2 mo, 70+/-8; 3 mo, 63+/-6; 4 mo, 87+/-9). Fresh bovine germ cells are capable of colonization in the busulfan-treated nude mouse testis, making it a suitable model for evaluation and development of spermatogonial transplant techniques in bulls.

Evaluation of American Wagyu sires for scrotal circumference by age and body weight.

Thirty-six percent of American Wagyu bulls do not meet the current minimum standards set by the Society of Theriogenology for the breeding soundness exam. In contrast, only 15% of bulls of domestic breeds do not meet the minimum standards. Scrotal circumference measurements of Wagyu are smaller than those of other breeds. The objective of this research was to describe scrotal circumference of Wagyu bulls as it relates to age and BW. The data set consisted of 190 Wagyu bulls housed at two locations. One hundred forty-one bulls constituted the first set of data (location 1); scrotal circumference was measured one to six times per bull aged between 13 and 70 mo. Ninety-four of the bulls underwent semen evaluation for motility and morphology. Forty-nine bulls constituted the data set for which scrotal circumference and BW was measured one to nine times per bull between 5 and 21 mo of age (location 2). Mean scrotal circumference of bulls within each age group was as follows: 12 to 14 mo, 29.8 0.2 cm (mean +/- SE); 15 to 17 mo, 31.8 +/- 0.2 cm; 18 to 20 mo, 32.9 +/- 0.3 cm; 21 to 24 mo, 31.8 +/- 0.5 cm; and > 24 mo, 35.5 +/- 0.2 cm. Both age and BW were highly correlated to scrotal circumference (r = 0.81 and 0.82, respectively). Within each age group, there were a percentage of bulls that did not meet the minimum standard for scrotal circumference set by the Society of Theriogenology. The percentages were as follows: 12 to 14 mo, 46%; 15 to 17 mo, 25%; 18 to 20 mo, 33%; 21 to 24 mo, 42%; and > 24 mo, 32%. Morphology and motility were > 50% each in 91% of the bulls between ages 12 and 20 mo at location 1. Based on these data, it is recommended that Wagyu bulls be evaluated with the breed-specific minimum standards for scrotal circumference of 26 cm from 12 to 14 mo, 29 cm from 15 to 17 mo, and 30 cm from 18 to 20 mo of age.

LHRH antagonist decreases LH and progesterone secretion but does not alter length of estrous cycle in heifers.

The objective of this study was to evaluate the suppressive effect of an LHRH antagonist, Cetrorelix SB-75 (SB-75), on secretion of LH, FSH and ovarian function in beef heifers. In Exp. 1, heifers were treated with a single dose of 10 microg/kg body weight intramuscularly on d 3 of the estrous cycle. In Exp. 2, heifers received either a single injection (100 microg/kg) of SB-75 on d 3 of the estrous cycle or multiple injections of 20 microg/kg on d 3, 4, 5, 6, and 7. Serum LH, but not FSH, was suppressed from one to several days. However, neither FSH nor progesterone was significantly altered. In Exp. 3, heifers received an injection vehicle (5% mannitol) or 100 microg/kg BW of SB-75 on d 1 of the estrous cycle (30 h after first observed standing estrus). Injection of SB-75 suppressed LH pulse frequency on d 3, 5, and 7 (P < 0.001). The mean LH concentrations in the SB-75 treatment groups were lower on d 3 (P < 0.01) and 5 (P < 0.05). There were no differences (P > 0.1) between the two groups in the mean concentrations of LH on d 1, 7, or 14. Treatment did not affect the secretion pattern or concentration of FSH. Injection of SB-75 did not alter estradiol-173 concentrations (P > 0.1). Treatment reduced corpus luteum (CL) function as indicated by lower progesterone production. However, the length of the estrous cycle was not shortened. These data show that the CL can form and survive in the face of depressed LH concentrations during the early stages of the estrous cycle.

The effects of recombinant LHRH fusion proteins on testicular development and histology in ram lambs.

Two recombinant luteinizing hormone releasing hormone (LHRH) fusion proteins were evaluated for their effectiveness in suppression of testicular development and histology by injecting together. Recombinant fusion proteins, ovalbumin-LHRH-7 and thioredoxin-LHRH-7, were generated using recombinant DNA technology and expressed in E. Coli. Eleven ram lambs ranked by age and body weight were randomly assigned to receive either ovalbumin and thioredoxin recombinant protein mixture (control group, n = 5) or ovalbumin-LHRH-7 and thioredoxin-LHRH-7 recombinant fusion protein mixture, anti-LHRH vaccine, (immunization group, n = 6). Animals in each group were weaned at 17 wk of age and were injected (primary immunization) with either mixture at 18 wk of age. Both groups received a booster immunization 8 wks later (26 wk of age). Scrotal circumference, scrotal length, testicular diameter and testicular length were measured in both groups every other week. All animals were slaughtered at 36 wk of age. Immediately after slaughter, a small testicular tissue was taken and processed for histological examination. In the ram lambs in immunization group scrotal circumference and testicular diameter increased steadily until second booster and then remained as a plateau until the end of the experiment. The differences in scrotal circumferences and testicular diameter were significant between the two groups during the last three weeks of the study (p < 0.05). There were no differences in testicular and scrotal length throughout the study (p > 0.05). Seminiferous tubules lost their regular shape and were decreased in diameter in immunization group. Although a few spermatozoa were seen in some tubules, in general, there were atrophy of the seminiferous tubules and loss of spermatogenesis, nevertheless, it seemed that animals in this group were potentially fertile.