Seasonal variation in freezability of buffalo semen

Fifty-six buffalo semen ejaculates of mass activity greater than plus three on a five-point scale (28 each in summer and winter seasons) were frozen in Tris yolk glycerol (TY-G) extender with four hours equilibration time. Motility of semen was checked after first extension (initial motility), after equilibration time, 15 minutes after freezing, and 30 days after freezing. Motility increased during winter season. Significant differences (P<0.01) in motility after freezing were observed between summer and winter seasons.     

Seasonal changes of semen quality and freezability in Franches-Montagnes stallions

The objective of this study was to investigate seasonal changes of semen quality parameters in Franches-Montagnes stallions and to compare the freezability of ejaculates collected in autumn and winter. Experiments were performed using 15 stallions from the National Stud Farm in Avenches (Switzerland). Ejaculates were collected and evaluated every month during 1 year as well as cryopreserved in autumn and winter (September to February). In fresh semen the gel-free volume, concentration, motility and morphology (normal sperm, major defects, vacuoles and acrosome defects) were evaluated and in frozen-thawed semen the motility as well as the viability (SYBR-14/PI) were performed. To analyse seasonal differences four periods of 3 months each were defined as autumn (September, October, November), winter (December, January, February), spring (March, April, May) and summer (June, July, August). During the 1-year experiment all fresh semen quality parameters demonstrated a clear seasonal and individual pattern. The gel-free volume was significantly (P<0.05) higher in spring and summer compared to autumn and winter while sperm concentration was significantly (P<0.05) lower in spring than at any other time of the year. Total sperm number was significantly (P<0.05) higher and sperm motility significantly (P<0.05) lower in summer than in other seasons. Regarding sperm morphology, normal sperm was significantly (P<0.05) higher in autumn than in winter and summer and major defects were lowest (P<0.05) in autumn. In frozen-thawed semen motility was significantly (P<0.05) improved in the ejaculates collected in autumn compared to winter, while viability showed no obvious differences. Our results clearly demonstrate that individual and seasonal differences occurred in semen quality of Franches-Montagnes stallions. Ejaculates collected in autumn (September, October, November) demonstrated good quality, especially regarding sperm morphology, and were more suitable for cryopreservation because of better motility in frozen-thawed semen collected during autumn than in winter.     

Seasonal changes in semen quality and freezability in the Warmblood stallion

The objective of this study was to investigate seasonal changes in stallion semen quality and to determine the best time for semen cryopreservation. Experiments were performed using 10 Warmblood stallions from the National Stud Farm in Avenches (Switzerland). Ejaculates were collected and frozen every other week during 1 year from January to December 1999. Volume, concentration, and motility, and the number of morphologically normal sperm and sperm with major defects (abnormal heads, acrosome defects, nuclear vacuoles, proximal droplets, abnormal midpieces) were evaluated. For all frozen-thawed semen samples motility as well as viability (SYBR-14/PI) was tested, and the hypoosmotic swelling test (HOS) was performed. To analyze seasonal differences 4 periods of 3 months each were defined: autumn (September, October, November), winter (December, January, February), spring (March, April, May) and summer (June, July, August). During the 1 year experiment all semen quality parameters showed a clear seasonal pattern. The volume, total sperm count and motility in fresh semen were significantly higher (P<0.05) in summer than in winter, while sperm concentration was significantly lower in summer compared to the other seasons. Regarding morphology, normal sperm was significantly lower (P<0.05) in summer than at any other time of the year and higher values (P<0.05) were found for major defects in summer than in spring and autumn. In frozen-thawed semen motility was significantly (P<0.05) improved in autumn when compared to spring and summer. Viability was lowest in summer and differed significantly (P<0.05) from other seasons. The HOS test revealed significantly more (P<0.05) membrane damaged spermatozoa in winter than in spring, summer and autumn. Our results demonstrate that in our climatic conditions clear seasonal differences occur in semen quality of fresh and frozen-thawed semen and that cryopreservation of stallion semen should preferably be performed in autumn.     

SDS-polyacrylamide gel electrophoresis of buffalo bulls seminal plasma proteins and their relation with semen freezability

The objective of this study was to evaluate the protein profiles of seminal plasma in buffalo bulls and to examine their correlation with semen characteristics. Semen of 10 buffalo bulls were collected by a bovine artificial vagina. Semen characteristics (motility, morphology, viability and concentration) were recorded. A part of the semen sample (1 ml) was diluted by tris-egg yolk-glycerol extender, packed in French straws and was frozen in liquid nitrogen. The straws were later thawed and semen characteristics were compared with those of the fresh semen. Seminal plasma was harvested by centrifugation; treated with cold ethanol and then, underwent SDS-polyacrylamide gel electrophoresis (PAGE). Twenty five protein bands were identified on the gel, of which those of <35.5 kDa were prominent (72% of the bands). Of these protein fractions, 24.5 kDa was significantly correlated with sperm progressive motility in fresh and viability in frozen-thawed semen while 45 kDa bands were correlated with abnormal morphology in frozen-thawed semen; 55 kDa protein fractions were correlated with sperm viability of fresh semen. Progressive motility, viability and abnormal sperm morphology of frozen-thawed semen were highly correlated with these parameters in the fresh semen. In conclusion, seminal plasma protein fractions in buffalo bulls are similar to those reported in other animal species and have some correlations with semen characteristics before and after freezing.     

Associations of scrotal circumference with semen traits in young beef bulls

Associations of percent live sperm, sperm number, sperm concentration, sperm motility, and potential breeding efficiency score with scrotal circumference were studied using data from the records of 465 Polled Hereford and 264 Simmental bulls. Data were subdivided by breed and within breed into three groups according to scrotal circumference (SC) size. Group I bulls had SC measurements <32 cm. Group II included bulls with SC measurements between 32 cm and the mean measurement of the breed (i.e., 35 cm for Polled Hereford and 38 cm for Simmentals). Group III was comprised of bulls with SC values above their respective breed averages. Age and body weight differences were also evaluated in the three groups. Average age differences among bulls in the three groups differed by not more than nine days in the two breeds. Average body weight differences among the three groups were statistically significant (P<0.05) with the maximum difference between any two groups within breed being 80 pounds. Average differences in percent live sperm and sperm number were greater (P<0.05) in Groups II and III than in Group I. For sperm concentration ratings, sperm motility ratings and potential breeding efficiency score, and for both breeds, Group I had a significantly lower (P<0.05) number of bulls classified satisfactory and a significantly greater (P<0.05) number of bulls classified unsatisfactory than did groups II and III.     

Estimates of non-genetic effects and genetic parameters for semen traits in Piemontese bulls

Male reproductive performances are often ignored in cattle breeding programmes, although semen traits might be used to improve bull breeding soundness. Effects of genetic and environmental factors on semen production and quality traits were estimated in 693 Piemontese bulls with the aim of providing the first estimates of genetic parameters for semen traits for this breed. Volume and concentrations of individual ejaculates (up to three per each test-day), and volume, concentration, total number of spermatozoa and post-thawing progressive motility of within test-day pooled semen were available for 19 060 ejaculates. Bulls reached the maximum amount of daily semen production after their third year of age, with concentration rapidly increasing until 23 months of age, and then slowly decreasing. Semen volume was at its highest when collection days were at least 15 days apart, whereas the maximum concentration was reached when the interval was 6 days. Heritability estimates were generally moderate (0.14–0.26), and low for progressive motility (0.08). Estimates of genetic correlation among the volumes of the individual ejaculates were high and positive (≥0.79), as were the genetic correlations among their concentrations (≥0.46). Genetic correlations among volume and concentration traits varied from −0.47 (with a 95% high posterior density interval ranging from −0.65 to −0.23) to −0.32 (with a 95% high posterior density interval ranging from −0.55 to −0.09). Progressive motility was unrelated with the other traits, but moderately positively correlated with volumes of the second and third ejaculates. The magnitude of heritabilities showed that selection for semen traits is possible. However, the unfavourable relationship between volume and concentration must be taken into account if a future selection programme is to be established.     

Seasonal variation in the semen characteristics of Muturu (Bos brachyceros) bulls

Semen was collected by electroejaculation from seven mature Muturu bulls at weekly intervals for 9 weeks in each of three seasons (‘Dry’, ‘Rainy’, and ‘Late Rainy/Early Dry’) to study the effect of season on semen characteristics. The respective mean values for the ‘Dry’, ‘Rainy’ and ‘Late Rainy/Early Dry’ seasons were: volume 1.8 ± 0.1, 2.3 ± 0.1 and 2.0 ± 0.1 ml; percentage motility 36.2 ± 2.6, 37.7 ± 2.8 and 27.5±2.9; and percent morphologically normal sperm 70.0 ± 3.1, 79.1 ± 2.6 and 79.8 ± 2.3. No seasonal differences were found for sperm concentration/ml (overall mean 1.92 ± 0.16 × 10⁸) and for sperm output/ejaculate (overall mean 4.14 ± 0.39 × 10⁸). The chemical composition of seminal plasma (mg %) for the respective seasons was calcium: 4.8 ± 1.0, 7.9 ± 0.9 and 2.5 ± 0.7; magnesium: 4.8 ± 0.4, 4.3 ± 0.7 and 2.5 ± 0.7; and chloride: 330.7 ± 33.7, 136.0 ± 9.6 and 344.3 ± 31.8. No seasonal differences were found in sodium or potassium concentrations. Fructose was found in the semen of only one bull and only during the ‘Dry’ season.     

Genetic correlations between yield traits or days open measured in cows and semen production traits measured in bulls

We used a bivariate animal model to investigate the genetic correlations between yield traits or days open (DO) as characters measured in cows and semen production traits as characters measured in bulls. Lactation records of 305-day milk, fat, and protein yields, and DO, from 386 809 first-lactation Holstein cows in Hokkaido, Japan, that calved between 2008 and 2014 were used. Semen production records were collected between 2005 and 2014 and included volume per ejaculate (VOL), sperm concentration (CON), number of sperm per ejaculate (NUM), progressive motility index of sperm (MOT), and MOT after freeze-thawing (A-MOT). Number of sperm per ejaculate was log-transformed into a NUM score (NUMS). A total of 30 373 semen production records from 1196 bulls were obtained. The pedigree file used for analysing the records was involving 885 345 animals. Heritability was estimated for VOL (0.42), CON (0.12), NUMS (0.37), MOT (0.08), and A-MOT (0.11). Weak and negative genetic correlations were recorded between yield traits measured in cows and VOL, CON or NUMS measured in bulls. Moderate and negative genetic correlations were obtained between DO and MOT (–0.42) or A-MOT (–0.43). Selection focused on MOT or A-MOT measured in bulls may therefore improve DO measured in cows.     

Testicular, semen and blood parameters in adapted and nonadapted Bos taurus bulls in the semi-arid tropics

Two experiments were conducted to evaluate differences in testicular, seminal and hematological characteristics in adapted (Senepol) and nonadapted (Holstein) Bos taurus bulls under the semi-arid environmental conditions of St. Croix, Virgin Islands (17 degrees N, 64 degrees W). In Experiment 1 mature, sexually-rested Senepol (n=10) and Holstein (n=9) bulls of similar age (37 months) and body weight (715 kg) and grazing on adjacent native pastures, were tested on the same day in July (28.8 degrees C mean ambient temperature, 81.5% humidity). Senepol bulls had lower (P<0.01) rectal temperatures (39.3 vs 40.0 degrees C) and higher (P<0.01) packed cell volumes (41.4 vs 35.2%) than Holstein bulls. Scrotal circumference was 3 cm larger (P<0.1) and testis tone firmer (P<0.001) in Senepol compared to Holstein bulls. Ejaculates, obtained by electroejaculation, contained 3.2x10(9) more spermatozoa with fewer abnormal tails and detached acrosomes in Senepol than in Holstein bulls (P<0.05). In Experiment 2, Senepol (n=42) and Holstein (n=30) bulls, representing 3 beef and 5 dairy farms, were evaluated during the summer (August/September) and winter (February/March). Again, scrotal circumference was larger (P<0.05) and testis tone firmer (P<0.001) in Senepol than in Holstein bulls, with no effect of season. Seminal fructose was higher (P<0.01) in Senepol than in Holstein bulls and decreased (P<0.01) during the winter collection. Blood plasma urea nitrogen and glucose were similar between breeds, but urea nitrogen was lower (P<0.01) during the summer. Significant (P<0.01) breed-by-age interactions were observed for the frequency of spermatozoa with protoplasmic droplets, decreasing in Holstein but not in Senepol bulls. The data point to differences between the adapted and nonadapted breed type in testicular and ejaculate characteristics, but also suggest that season has only a limited impact on bull reproductive function under the environmental conditions in St. Croix.     

Equine frozen semen: freezability and fertility field results

The freezability of stallion semen defined as the number of selected ejaculates/total number of ejaculates frozen from 161 different stallions was analyzed. Of the stallions, 19, 30, 27 and 24% had a freezability of 0%, 0 to 33%, 33 to 66%, over 66%, respectively In 85 different stallions, the correlation of freezability between first and second year was 0.60 (P < 0.001). The relationship between fertility with fresh and frozen semen and freezability was analyzed in 40 stallions whose freezability and fertility information was recorded during 5 years. There was a strong relationship between fertility of fresh semen and semen freezability (P < 0.001). However, the relationship between fertility of frozen semen and freezability was not as marked (P < 0.05). Analysis of the field fertility per cycle results when mares were bred with 300 or 150 x 10(6) total spermatozoa at different frequencies until ovulation indicated that mares that were inseminated 2 times or more per estrus show an improved fertility in comparison with mares inseminated only once (34%, n = 1576 vs 26%, n = 626; P < 0.001). Foaling rate when mares were inseminated with frozen semen (1858 mares during 8 breeding seasons) was mainly influenced by mare age (< 16 years: 54% vs >/= 16 years 42% p < 0.001). Date of first insemination (before May 15: 58% vs after May 15: 37%) also had a significant effect on foaling rate (P < 0.001).     

Genetic and phenotypic parameter estimates for scrotal circumference and semen traits in young beef bulls

Estimated in this study were heritabilities and genetic and phenotypic correlations involving scrotal circumference (SC), percent live sperm, sperm number, sperm concentration, sperm motility, and an overall measure of a bull's potential breeding efficiency. Potential breeding efficiency is a composite trait based on a consideration of sperm concentration, sperm motility, sperm morphology and scrotal circumference. Data used were from three sources. Records on 863 Angus, 753 Polled Hereford, and 302 Simmental bulls were made available through the Missouri Performance-Tested Bull Sale and records on 1169 Polled Hereford bulls came from the American Polled Hereford Association. Information from these first two data sets were used to estimate heritability of scrotal circumference. The third data set was provided by Nichols Farms of Bridgewater, Iowa, and included information from the records of 465 yearling Polled Hereford and 264 yearling Simmental bulls. This latter data set was used to estimate all of the above mentioned parameters. Each data set was kept separately for the purpose of statistical analysis. Parameters were estimated using components from paternal half-sib analysis of variance and covariance. Pooled estimates of heritability for SC, sperm concentration, sperm motility, percent live sperm, sperm number and potential breeding efficiency were 0.51 +/- 0.09, 0.20 +/- 0.13, 0.11 +/- 0.12, 0.00, 0.19 +/- 0.14 and 0.13 +/- 0.12, respectively. Phenotypic correlations involving the six traits were very consistent for the two breeds. Combined across breeds their values ranged from 0.47 for SC and percent live sperm to 0.96 for sperm concentration and potential breeding efficiency. Corresponding genetic correlations were generally positive and high and ranged from 0.65 for SC and sperm motility to 1.14 for sperm number and potential breeding efficiency.     

Seasonal changes in semen quality and correlation with plasma hormone profiles in Karan Fries bulls

The aim of the study was to assess the influence of summer and winter seasons on semen quality and plasma hormone concentrations in cross-bred bulls. Semen was collected by an artificial vagina from eight bulls and microscopically evaluated for quality parameters. Semen volume was higher in summer season (p < 0.05) than winter season, whereas nonsignificant variation (p > 0.05) was observed in mass motility, individual motility, sperm viability, sperm concentration and percentage of membrane-intact and acrosome-intact spermatozoa. Plasma prolactin and testosterone concentration were significantly (p < 0.01) higher in summer season than winter season. Plasma testosterone levels were positively correlated with semen volume and negatively correlated with individual motility (p < 0.05). Prolactin showed a significant positive correlation with semen volume. A well-defined seasonal pattern in semen characteristics was not observed and few correlations existed between plasma hormone levels and semen characteristics in Karan Fries bulls.     

The effect of gossypol in cottonseed meal on performance and on hematological and semen traits in postpubertal Brahman bulls

The effects of a cottonseed meal diet containing high levels of free gossypol on hematological traits, including erythrocyte osmotic fragility and semen characteristics, were examined during an 11-week period. Eight Brahman bulls were randomly assigned to 1 of 2 groups. The control group (n=4) was fed a mixture of soybean meal and corn. The treated group (n=4) was fed a mixture of cottonseed meal and corn. Both groups were allowed hay free choice. The treated group consumed 8.2 g of free gossypol per bull per day. The percentage of normal spermatozoa was lower (P<0.01) in the treated than in the control group from Week 5 (49+/-9.8 vs 83+/-3.2%), which was primarily influenced by changes in mid piece morphology in the treated bulls. Erythrocyte osmotic fragility was higher (P<0.001) in the treated than in the control group over the entire study period, although group values diverged more acutely from Week 7 of treatment. Sperm motility was lower (P=0.04) in treated bulls than in control bulls at Week 9 (52+/-9.8 vs 82+/-6.2%). These data suggest that concurrent discernable changes in bull erythrocyte osmotic fragility and in semen characteristics can occur following commencement of a diet containing 8.2g of free gossypol per day.     

Influence of repeated treadmill exercise on quality and freezability of stallion semen

The objective of this study was to investigate changes of quality and freezability of stallion semen in response to repeated acute treadmill exercise. Ejaculates from 11 stallions were collected, evaluated and frozen weekly during four periods of 4 weeks each defined as before (period 1), during (period 2) and after (periods 3 and 4) intense exercise. In fresh semen the gel-free volume, sperm concentration, motility, normal sperm and sperm with major defects (acrosome defects, nuclear vacuoles, abnormal heads, midpiece defects and proximal droplets) were evaluated. In frozen-thawed semen, motility as well as viability (SYBR-14/PI) were examined. In period 2, all stallions were exercised on an indoor high speed treadmill twice a week (total of eight sessions) using an incremental workload test. Heart rate was monitored telemetrically during exercise and blood samples were taken for determination of cortisol, testosterone and lactate. Results of our investigation demonstrate that heart rate and the plasma concentrations of cortisol, testosterone and lactate significantly (P < 0.05) increased during each exercise session. Furthermore, significantly more major sperm defects were present in periods 3 (69.5+/-2.1%) and 4 (66.8+/-2.1%) than in periods 1 (62.2+/-2.4%) and 2 (62.5+/-2.2%). Acrosome defects increased towards the end of exercise but improved 3 weeks later to values observed before exercise. In frozen-thawed semen, motility was significantly lower in period 2 (45.4+/-2.3%) compared to period 4 (51.6+/-1.7%) and viability was significantly lower in period 2 (49.2+/-2.0%) than in periods 1 (53.8+/-2.1%) and 4 (53.7+/-1.6%). Our results clearly demonstrate that in the stallion repeated strenuous treadmill exercise can negatively influence semen quality and freezability.     

Seasonal variation in semen quality in Bos indicus and Bos taurus bulls raised under tropical conditions

In the present study, we tested the hypothesis that Bos taurus taurus bulls have greater reactive oxygen species (ROS) and lower activity of antioxidant enzymes in their semen than Bos taurus indicus bulls. Sixteen Simmental bulls (B. t. taurus) and 11 Nelore bulls (B. t. indicus) were managed extensively in a tropical environment. Semen was collected twice annually (summer and winter) for 2 consecutive years. Simmental bulls had significantly higher percentages of major sperm defects during the summer than the winter (20.3+/-3.1% versus 12.2+/-2.4%, respectively; mean+/-S.E.M.). There was an interaction of breed and season for minor sperm defects (P=0.037; highest in Nelore bulls in the summer) and an effect of season on total defects (P=0.066; higher in summer). To evaluate oxidative damage, malondialdehyde (lipid-peroxidation metabolite) concentrations were indirectly measured by semen concentrations of thiobarbituric acid reactive substances (TBARS); these were higher in summer than in winter (728.1+/-79.3ng/mL versus 423.8+/-72.6ng/mL, respectively; P=0.01). Glutathione peroxidase/redutase (GPx) activity in semen was higher in Simmental versus Nelore bulls (741.6+/-62.1 versus 510.2+/-62.8; P<0.01). However, superoxide dismutase (SOD), another antioxidant enzyme, was not significantly affected by breed or season. There were correlations between TBARS and sperm primary defects during the summer for both Simmental and Nelore bulls (r=0.59, P=0.021 and r=0.40, P=0.034, respectively), and between SOD and primary defects during summer for Simmental bulls only (r=-0.51, P=0.041). In conclusion, there was a higher level of lipid peroxidation (ROS) in semen of Simmental versus Nelore bulls; apparently the higher GPx activity in Simmental bulls was insufficient to avoid damage that occurred concurrent with increased ROS production during the summer.     

Influence of vedaprofen (Quadrisol) on quality and freezability of stallion semen

The objective of this study was to evaluate the effect of the non-steroidal anti-inflammatory drug (NSAID) vedaprofen (Quadrisol) on quality and freezability of stallion semen. Experiments were performed using 22 Franches Montagnes stallions from the National Stud in Avenches (Switzerland) randomly divided into a control and test group. Vedaprofen was given orally to all stallions of the test group at the recommended therapeutic dose (initial dose of 2mg/kg followed by 1mg/kg body weight every 12h) for 14 days. Control animals received the same amount of carrier substance. During treatment, blood samples of five stallions in both test and control group were collected for PGF(2 alpha)-metabolite (PG-metabolite) determination. Ejaculates from all stallions were collected and cryopreserved weekly for 14 weeks from September to December. Concentrations of PG-metabolite, PGF and PGE were measured in the seminal plasma of ejaculates collected 2 weeks before, during and 2 weeks after treatment. In fresh semen the volume, concentration, motility and number of normal sperm and sperm with major defects (acrosome defects, abnormal heads, nuclear vacuoles, proximal droplets, midpiece defects) were evaluated. In frozen-thawed semen samples motility as well as viability (SYBR-14/PI) were tested and the hypoosmotic swelling test (HOS) was performed. Results demonstrate that vedaprofen had no effect on blood plasma concentration of PG-metabolite but significantly inhibited both, PGF and PGE concentrations in seminal plasma. Furthermore, all quality parameters in fresh and frozen-thawed semen were not affected by vedaprofen treatment but the time of semen collection had a significant (P<0.05) effect on motility, normal sperm and sperm with nuclear vacuoles in fresh semen.     

Genome-wide detection of non-additive quantitative trait loci for semen production traits in beef and dairy bulls

Semen production traits are important aspects of bull fertility, because semen quantity leads to direct profits for artificial insemination centres, and semen quality is associated with the probability of achieving a pregnancy. Most genome-wide association studies (GWASs) for semen production traits have assumed that each quantitative trait locus (QTL) has an additive effect. However, GWASs that account for non-additive effects are also important in fitness traits, such as bull fertility. Here, we performed a GWAS using models that accounted for additive and non-additive effects to evaluate the importance of non-additive effects on five semen production traits in beef and dairy bulls. A total of 65 463 records for 615 Japanese Black bulls (JB) and 50 734 records for 873 Holstein bulls (HOL), which were previously genotyped using the Illumina BovineSNP50 BeadChip, were used to estimate genetic parameters and perform GWAS. The heritability estimates were low (ranged from 0.11 to 0.23), and the repeatability estimates were low to moderate (ranged from 0.28 to 0.45) in both breeds. The estimated repeatability was approximately twice as high as the estimated heritability for all traits. In this study, only one significant region with an additive effect was detected in each breed, but multiple significant regions with non-additive effects were detected for each breed. In particular, the region at approximately 64 Mbp on Bos taurus autosome 17 had the highest significant non-additive effect on four semen production traits in HOL. The rs41843851 single nucleotide polymorphism (SNP) in the region had a much lower P-value for the non-additive effect (P-value = 1.1 × 10^?31) than for the additive effect (P-value = 1.1 × 10^?8) in sperm motility. The AA and AB genotypes on the SNP had a higher phenotype than the BB genotype in HOL, and there was no bull with the BB genotype in JB. Our results showed that non-additive QTLs affect semen production traits, and a novel QTL accounting for non-additive effects could be detected by GWAS. This study provides new insights into non-additive QTLs that affect fitness traits, such as semen production traits in beef and dairy bulls.