Anatomical studies on the PES region of Zebu cattle (Bos Taurus indicus) with special references to 3D computed tomography imaging technique.

The 3D render volume reconstruction CT (3D-RVCT) produced detailed images of the PES region, determining its relationships with the surrounding structures. Despite extensive research in veterinary studies on the PES through gross anatomy and CT, there is a lack of studies on the PES of zebu cattle. The study aimed to analyze the PES of Zebu cattle using gross cross-sectional, radiographic, CT, and morphometric methods, with the use of 3D-RVCT to provide anatomical guidance for surgeons and students. The study was performed on sixteen PES regions to provide hard and soft tissues in CT images. Three are five tarsal bones and two large fused (III and IV) metatarsal bones that were completely fused except for their distal extremities, which were divided distally by the intertrochlear notch. The cortical thickness of the metatarsal bone was equal on both sides. The bony septum divided the medullary cavity between the two fused large metatarsal bones in the proximal distal half only and disappeared in the middle part. The reconstruction showed similar sizes in the right and left limbs, confirming the pes bones. The radiographic and CT images could be used as a normal reference for the interpretation of some clinical diseases in the PES. The 3D CT reconstruction of the pes bones was described by various CT oblique dorsal and plantar views. The study focuses on diagnosing PES disorders using CT imaging, improving medical interventions, improving Zebu cattle health outcomes, and empowering students to contribute to veterinary medicine research and advancements.

Effect of exogenous melatonin implant on post-thaw semen quality of buffalo bulls.

Melatonin is an intracellular antioxidant of sperm membrane that protects the cells from lipid peroxidation. Yet, its role as an antioxidant on semen quality of buffalo bulls is still obscure. The present study was undertaken to assess the effect of exogenous melatonin implant (18 mg/50 kg bodyweight) on post-thaw sperm characteristics, oxidative stress, endocrinological profiles and fertility of buffalo bulls. Six apparently healthy breeding Murrah buffalo bulls were randomly selected at bull farm, Guru Angad Dev Veterinary and Animal Sciences University for the present study and divided into two groups viz. control (n = 3) and melatonin implanted group (n = 3). A total of 120 ejaculates were collected from bulls of both groups (n = 60 each) throughout the study period. Most beneficial effects of melatonin implants were observed during post-implantation period. The percentages of post-thaw sperm total and progressive motility, viability and mitochondrial membrane potential were higher (p < .05) in melatonin implanted buffalo bulls compared to controls during post-implantation period. Following melatonin implantation, MDA production in post-thaw semen was lower (p < .05) in melatonin implanted group than in control group. Plasma melatonin and testosterone concentrations were higher (p < .05) in buffalo bulls implanted with melatonin as compared to their control counterparts. No differences (p > .05) in plasma LH concentrations were observed in both groups. First service pregnancy rate was 43.3% using semen of melatonin implanted bulls and 30.0% with semen of controls (p > .05). Thus, melatonin was able to protect sperm membrane against oxidative damage and improve post-thaw semen quality, thereby resulting in higher fertilizing potential of spermatozoa.

Kaempferol as an Alternative Cryosupplement for Bovine Spermatozoa: Cytoprotective and Membrane-Stabilizing Effects.

Kaempferol (KAE) is a natural flavonoid with powerful reactive oxygen species (ROS) scavenging properties and beneficial effects on ex vivo sperm functionality. In this paper, we studied the ability of KAE to prevent or ameliorate structural, functional or oxidative damage to frozen-thawed bovine spermatozoa. The analysis focused on conventional sperm quality characteristics prior to or following thermoresistance tests, namely the oxidative profile of semen alongside sperm capacitation patterns, and the levels of key proteins involved in capacitation signaling. Semen samples obtained from 30 stud bulls were frozen in the presence of 12.5, 25 or 50 µM KAE and compared to native ejaculates (negative control-CtrlN) as well as semen samples cryopreserved in the absence of KAE (positive control-CtrlC). A significant post-thermoresistance test maintenance of the sperm motility (p < 0.001), membrane (p < 0.001) and acrosome integrity (p < 0.001), mitochondrial activity (p < 0.001) and DNA integrity (p < 0.001) was observed following supplementation with all KAE doses in comparison to CtrlC. Experimental groups supplemented with all KAE doses presented a significantly lower proportion of prematurely capacitated spermatozoa (p < 0.001) when compared with CtrlC. A significant decrease in the levels of the superoxide radical was recorded following administration of 12.5 (p < 0.05) and 25 µM KAE (p < 0.01). At the same time, supplementation with 25 µM KAE in the cryopreservation medium led to a significant stabilization of the activity of Mg2+-ATPase (p < 0.05) and Na+/K+-ATPase (p < 0.0001) in comparison to CtrlC. Western blot analysis revealed that supplementation with 25 µM KAE in the cryopreservation medium prevented the loss of the protein kinase A (PKA) and protein kinase C (PKC), which are intricately involved in the process of sperm activation. In conclusion, we may speculate that KAE is particularly efficient in the protection of sperm metabolism during the cryopreservation process through its ability to promote energy synthesis while quenching excessive ROS and to protect enzymes involved in the process of sperm capacitation and hyperactivation. These properties may provide supplementary protection to spermatozoa undergoing the freeze-thaw process.

Effects of hyaluronic acid on sperm parameters, mitochondrial function and apoptosis of spermatozoa in Simmental bulls with good and poor freezing ability.

Bulls with varying freezability exhibit substantial variation in semen characteristics after cryopreservation. Sperm freezability is positively correlated with membrane cholesterol content, membrane integrity, mitochondrial activity and antioxidant content. The purpose of this study was to determine the optimal concentration of hyaluronic acid (HA) in bull sperm with different cryotolerances. Simmental bulls (n = 10) semen samples were taken and categorized based on their progressive motility (PM) after freeze-thawing: Group I, consisting of bulls (n = 5) with progressive sperm motility ≥45%, was considered good freezability ejaculates (GF), and Group II, including bulls (n = 5) with progressive sperm motility ≤30%, was considered poor freezability ejaculates (PF) bulls. Semen samples were diluted with a Tris-egg-yolk-glycerol (TEYG) extender containing various concentrations of HA: without HA (control), 1 mM HA, 2 mM HA and 4 mM HA. After the freeze-thaw process, sperm kinematics, plasma membrane and acrosome integrity, mitochondrial activity and apoptotic status were evaluated. The addition of 1 mM HA to the diluent of bulls with GF increased PM and linearity (LIN) compared to the control group (p < 0.05). Normal morphology was improved after thawing in the samples treated with 1 and 2 mM HA in the GF and PF bulls respectively. The membrane and acrosome integrity of GF bulls treated with 1 mM HA was significantly (p < 0.05) greater than that of the control groups. Adding 1 mM HA to the extender of bulls with GF and PF improved the proportion of viable cells compared with the highest concentration (4 mM) of HA. The mitochondrial activity of PF bulls treated with 1 and 2 mM HA was significantly (p < 0.05) greater than that of the controls and 4 mM HA. Finally, it can be concluded that adding low doses of HA (1 mM) to the TEYG extender of GF and PF bulls ameliorated the post-thaw semen quality.

Effects of folic acid and riboflavin on growth performance, nutrient digestion and rumen fermentation in Angus bulls.

This study examined the influences of coated folic acid (CFA) and coated riboflavin (CRF) on bull performance, nutrients digestion and ruminal fermentation. Forty-eight Angus bulls based on a randomised block and 2 × 2 factorial design were assigned to four treatments. The CFA of 0 or 6 mg of folic acid/kg DM was supplemented in diets with CRF 0 or 60 mg riboflavin (RF)/kg DM. Supplementation of CRF in diets with CFA had greater increase in daily weight gain and feed efficiency than in diets without CFA. Supplementation with CFA or CRF enhanced digestibility of DM, organic matter, crude protein, neutral-detergent fibre and non-fibre carbohydrate. Ruminal pH and ammonia N content decreased and total volatile fatty acids concentration and acetate to propionate ratio elevated for CFA or CRF addition. Supplement of CFA or CRF increased the activities of fibrolytic enzymes and the numbers of total bacteria, protozoa, fungi, dominant fibrolytic bacteria and Prevotella ruminicola. The activities of α-amylase, protease and pectinase and the numbers of Butyrivibrio fibrisolvens and Ruminobacter amylophilus were increased by CFA but were unaffected by CRF. Blood concentration of folate elevated and homocysteine decreased for CFA addition. The CRF supplementation elevated blood concentrations of folate and RF. These findings suggested that CFA or CRF inclusion had facilitating effects on performance and ruminal fermentation, and combined addition of CFA and CRF had greater increase in performance than CFA or CRF addition alone in bulls.

Identification of genomic regions associated with total and progressive sperm motility in Italian Holstein bulls.

Sperm motility is directly related to the ability of sperm to move through the female reproductive tract to reach the ovum. Sperm motility is a complex trait that is influenced by environmental and genetic factors and is associated with male fertility, oocyte penetration rate, and reproductive success of cattle. In this study we carried out a GWAS in Italian Holstein bulls to identify candidate regions and genes associated with variations in progressive and total motility (PM and TM, respectively). After quality control, the final data set consisted of 5,960 records from 949 bulls having semen collected in 10 artificial insemination stations and genotyped at 412,737 SNPs (call rate >95%; minor allele frequency >5%). (Co)variance components were estimated using single trait mixed models, and associations between SNPs and phenotypes were assessed using a genomic BLUP approach. Ten windows that explained the greatest percentage of genetic variance were located on Bos taurus autosomes 1, 2, 4, 6, 7, 23, and 26 for TM and Bos taurus autosomes 1, 2, 4, 6, 8, 16, 23, and 26 for PM. A total of 150 genes for TM and 72 genes for PM were identified within these genomic regions. Gene Ontology enrichment analyses identified significant Gene Ontology terms involved with energy homeostasis, membrane functions, sperm-egg interactions, protection against oxidative stress, olfactory receptors, and immune system. There was significant enrichment of quantitative trait loci for fertility, calving ease, immune response, feed intake, and carcass weight within the candidate windows. These results contribute to understanding the architecture of the genetic control of sperm motility and may aid in the development of strategies to identify subfertile bulls and improve reproductive success.

Seasonal and climatic factors have a significant influence on fertility associated sperm phenomic attributes in crossbred breeding bulls (Bos taurus × Bos indicus).

Although seasonal variations in semen quality and fertility have been studied to a considerable extent in breeding bulls, the effect of climatic variables on sperm functional competency has not been understood in detail. The present study analyzed sperm functional parameters in breeding bulls, over a period of 1 year, and assessed the effect of climatic variables on fertility associated sperm parameters. Seasons were categorized into summer, rainy, autumn, and winter based on the meteorological data. Semen was collected from crossbred bulls (n = 7) across the seasons and evaluated for functional membrane integrity, acrosome reaction status, protamine deficiency, capacitation, and lipid peroxidation status using specific fluorescent probes. The results of the present study revealed that bulls produced higher (p < 0.05) viable and acrosome intact spermatozoa during the autumn. The proportion of uncapacitated spermatozoa was also higher (p < 0.05) during autumn. Further, correlation of sperm functional attributes with environmental variables revealed that sperm viability was significantly (p < 0.05) and negatively correlated with daylength and temperature; acrosomal integrity was significantly (p < 0.05) and negatively correlated with day length; and protamine deficiency had significant (p < 0.05) positive correlation with day length and average temperature, and negative correlation with relative humidity. It was concluded that semen produced during autumn was superior to the semen produced during other seasons in terms of sperm functional competencies required for fertility.

Changes in genomic inbreeding and diversity over half a century in Swedish Red and Swedish Holstein dairy cattle.

Swedish Red (SR) and Swedish Holstein (SH) are the dominating commercial dairy cattle breeds in Sweden. Both breeds have undergone substantial changes during the last half century due to intensive selection for breeding goal traits, but also resulting from increased international exchange of breeding animals and genetic drift. The aim of this study was to learn more about changes in genomic diversity and inbreeding in these two breeds over time. Therefore, semen samples from old bulls were genotyped using the 150K Genomic Profiler SNP array and combined with 50K SNP array genotype data, obtained for more recent bulls from the Nordic Cattle Genetic Evaluation. Different measures of level of homozygosity, genomic inbreeding, relatedness and changes in allele frequency were estimated for bulls born during different time periods from the 1950s until 2020. In total, more than 33,000 SNPs for 9737 SR and 5041 SH bulls were included in the analysis using PLINK v1.9. The results showed higher average homozygosity for SR than for SH bulls up to around 2000, but the difference was very small after that. The average inbreeding coefficients based on deviation from expected homozygosity as well as on runs of homozygosity decreased until the early 1980s in both breeds, whereafter they started to increase again for SH, but stayed more stable for SR. From the 1990s onwards, SH displayed higher average inbreeding coefficients than SR. In the last studied birth year group (2015-2020), the mean inbreeding coefficient based on runs of homozygosity was 5.9% for SH and 3.7% for SR. A principal component analysis showed a pattern of genetic relationships related to the birth year period of the bulls, illustrating the gradual change of the genetic material within each breed. The change in allele frequency over time was generally larger for SH than for SR. The results show that the inbreeding level was higher half a century ago than at present, and the inbreeding levels were lower than in some other studied populations. Still, the increase seen for inbreeding coefficients and homozygosity, especially in SH during recent years, should be considered in future breeding strategies.

Epicatechin Prevents Cryocapacitation of Bovine Spermatozoa through Antioxidant Activity and Stabilization of Transmembrane Ion Channels.

Epicatechin (EPC) is a flavonoid belonging to the family of catechins; it has been described as a powerful scavenger of a wide spectrum of reactive oxygen species (ROS) and a modulator of ex vivo sperm vitality. In this study, we assessed the potential protective abilities of EPC on cryopreserved bovine spermatozoa. We focused on conventional quality parameters, as well as the oxidative profile of spermatozoa alongside capacitation patterns, and expression profiles of proteins involved in the process of capacitation. Semen samples were cryopreserved in the presence of 25, 50 or 100 µmol/L EPC and compared to native semen (negative control) as well as ejaculates frozen in the absence of EPC (positive control). A dose-dependent improvement of conventional sperm quality parameters was observed following EPC administration, particularly in case of the sperm motility, membrane, acrosome and DNA integrity in comparison to the positive control. Experimental groups exposed to all EPC doses presented with a significantly lower proportion of capacitated spermatozoa as opposed to the positive control. While no significant effects of EPC were observed in cases of superoxide production, a significant decrease in the levels of hydrogen peroxide and hydroxyl radical were recorded particularly in the experimental groups supplemented with 50 and 100 µmol/L EPC. Western blot analysis revealed that supplementation of particularly 100 µmol/L EPC to the semen extender prevented the loss of the cation channel of sperm (CatSper) isoforms 1 and 2, sodium bicarbonate cotransporter (NBC) and protein kinase A (PKA), which play important roles in the process of sperm capacitation. In summary, we may hypothesize that EPC is particularly effective in the stabilization of the sperm membrane during the freeze-thaw process through its ability to quench ROS involved in damage to the membrane lipids and to prevent the loss of membrane channels crucial to initiate the process of sperm capacitation. These attributes of EPC provide an additional layer of protection to spermatozoa exposed to low temperatures, which may be translated into a higher post-thaw structural integrity and functional activity of male gametes.

Mineral and glycerol concentrations in drinking water on body weight loss and acid-base balance in feed-deprived Holstein bulls.

Situations in which cattle are feed-deprived over extensive periods of time are common in the context of transport and is an animal welfare concern which may also compromise growth and carcass yield grade. This study investigated how the main components of an oral rehydration solution would affect BW loss and blood parameters in feed-deprived bulls. Three dose-response experiments were performed with 24 bulls each (n = 6 per treatment) to study the effect of mineral concentration in Study I (0, 100, 200 and 300 mOsm/kg), the K+ to Na+ ratio in Study II (25:75, 40:60, 60:40 and 75:25), and glycerol concentration in Study III (0%, 1%, 2% and 4% of the final solution). The blocking factor was initial BW and treatments were randomly assigned within each block. Measurements included fluid intakes, BW, and blood parameters at 0, 24 and 48 h relative to the start of feed deprivation. In Study I, increasing mineral concentration in solution linearly decreased BW losses at 48 h. At 24 and 48 h, serum urea linearly decreased with increasing mineral concentration. At 48 h, blood K+ and Na+ linearly increased, resulting in increased blood osmolarity. Additionally, at 24 h feed deprivation, blood pH linearly increased with increasing osmolality. In Study II, BW losses decreased with increasing K+ to Na+ ratio at 24 h, but not at 48 h. No effect of the K+ to Na+ ratio was found on blood parameters, apart from a trend for a linear decrease of blood osmolarity at 48 h. In Study III, serum urea tended to linearly decrease with increasing glycerol inclusion at 24 h, while blood glucose linearly increased with glycerol inclusion at 24 and 48 h. These combined results indicated that a solution with an osmolality of 200 mOsm/kg and a high K+ to Na+ ratio would effectively mitigate BW losses and maintain blood acid-base balance, whereas high glycerol inclusion sustained blood glucose in feed-deprived bulls.

Genetic analysis for semen quality traits in buffalo bulls.

This study was attempted to estimate the genetic parameters of semen quality traits in buffalo bulls. The study data consisted of 10975 ejaculates from 45 Murrah buffalo bulls (aged 24-72 months) used for breeding program during year 2010 to 2020. Semen quality traits (ejaculate volume, concentration of sperm, mass activity, initial and post-thaw motility, number of sperms per ejaculate, motile sperm number and discard rates) were studied. It was observed that average ejaculate volume was 2.82 ± 1.45 mL with mean concentration of 1040.12 ± 523.26 million/mL. Higher heritability was observed for number of sperms per ejaculate, number of motile sperm and sperm concentration. Significant phenotypic correlation was obtained between volume and number of sperms per ejaculate as well as volume and number of motile sperms. Likewise, significant phenotypic correlation was evident between sperm concentration with sperm number per ejaculate. Highest phenotypic correlation was obtained between sperm count per ejaculate and motile sperm count. Estimated genetic trends showed significant change in volume and motile sperm number. In conclusion, this study ascertains that genetic parameters of semen traits can be considered during the selection of buffalo bulls in breeding program.

Deciphering metabolomic alterations in seminal plasma of crossbred (Bos taurus X Bos indicus) bulls through comparative deep metabolomic analysis.

The incidence of sub-fertility is higher in crossbred bulls compared to zebu bulls. In the present study, we analysed the metabolomic profile of seminal plasma from crossbred and zebu bulls and uncovered differentially expressed metabolites between these two breeds. Using a high-throughput LC-MS/MS-based approach, we identified 990 and 1,002 metabolites in crossbred and zebu bull seminal plasma respectively. After excluding the exogenous metabolites, we found that 50 and 68 putative metabolites were unique to crossbred and zebu bull seminal plasma, respectively, whilst 87 metabolites were common to both. After data normalisation, 63 metabolites were found to be dysregulated between crossbred and zebu bull seminal plasma. Observed pathways included Linoleic acid metabolism (observed metabolite was phosphatidylcholine) in crossbred bull seminal plasma whereas inositol phosphate metabolism (observed metabolites were phosphatidylinositol-3,4,5-trisphosphate/inositol 1,3,4,5,6-pentakisphosphate/myo-inositol hexakisphosphate) was observed in zebu bull seminal plasma. Abundance of Tetradecanoyl-CoA was significantly higher, whilst abundance of Taurine was significantly lower in crossbred bull seminal plasma. In conclusion, the present study established the seminal plasma metabolomic profile in crossbred and zebu bulls and suggest that increased lipid peroxidation coupled with low concentrations of antioxidants in seminal plasma might be associated with high incidence of sub-fertility in crossbred bulls.

Genom-wide analysis identifies single nucleotide polymorphism variations and altered pathways associated with poor semen quality in breeding bulls.

The reason for poor semen quality among the breeding bulls is not well understood. In the present study, we performed high-throughput RNAseq analysis of spermatozoa to identify the SNPs present in good and poor-quality semen-producing Holstein Friesian breeding bulls. A total of 21,360 and 44,650 SNPs were identified in good and poor-quality semen with a minimum read depth of 20, among which 4780 and 8710 novel variants were observed in good and poor-quality semen, respectively. Greater SNPs and indels variations were observed in poor compared to good-quality semen. In poor-quality semen, SNP variations were observed in ZNF280B, SLC26A2, DMXL1, OR52A1, MACROD2 and REV1 genes, which are associated with regulation of spermatogenesis, post-testicular maturation, Cl- channel activity, V-ATPase-mediated intracellular vesicle acidification, a mono-ADP-ribosyl hydrolase and ATR-Chk1 checkpoint activation. GO analysis of filtered genes with significant variations between good and poor-quality semen showed enrichment in important pathways related to semen quality such as MAPK signalling pathway, Akt signalling pathway, focal adhesion, cAMP signalling pathway, and Rap1 signalling pathway. Network analysis of filtered genes in poor-quality semen showed variations in pathways of purine metabolism, pyrimidine metabolism, prolactin signalling pathway and RNA cap-binding complex. It is inferred that SNP in genes involved in maintaining sperm functions could be the reason for poor-quality semen production in bulls, and the identified SNPs hold potential to be used as biomarkers for semen quality in bulls.

Influence of tropical, subtropical, and temperate climate conditions on seminal redox status and sperm quality parameters in breeding bulls.

This study evaluated the effect of climate change on andrological parameters of beef bulls raised under tropical, subtropical, and temperate conditions. Bull ejaculates were collected to evaluate seminal quality parameters, sperm membrane integrity, and redox status (SOD; GPx; GSH; GRx; CARB; DCF; and SOD/GPx ratio). Bulls located in the temperate region showed a higher sperm motility rate and percentage of viable sperm (P < 0.05). When evaluating regions independently, we observed a lower GPx activity from animals in the tropical region (P < 0.05). In contrast, we found that SOD and GRx activities, GSH content, and CARB oxidative levels were higher in the tropical region, while oxidation values of DCF were lower (P < 0.05). Braford bulls showed higher CARB and DCF levels (1.23 ± 0.61 nmol/mg and 1453.60 ± 828.63 nmol/mg, respectively) compared to Hereford bulls (1.00 ± 0.43 nmol/mg and 1138.70 ± 423.24 nmol/mg, respectively) in the temperate region. However, Nellore bulls showed higher DCF levels (650.50 ± 401.53 nmol/mg) than Braford bulls (409.40 ± 286.97 nmol/mg). In addition, the SOD/GPx ratio was lower in Braford (12.44 ± 7.64 U/mg) compared to Nellore bulls in tropical conditions (87.25 ± 2.83 U/mg). A positive correlation was found in temperate conditions between DCF levels, SOD, and GRx activities (0.51, 0.58; P < 0.01, respectively), as well as in subtropical conditions between DCF levels and GRx activity (0.53; P < 0.01). A negative correlation between the temperature-humidity index and CARB content was found in subtropical and tropical regions (-0.44; P < 0.01). We concluded that Braford breeds showed lower seminal motility, DCF contents and SOD/GPx ratios compared to Nellore bulls in tropical climate conditions. Finally, in temperate environmental conditions, Braford bulls also showed lower seminal motility but higher levels of CARB and DCF contents compared to Hereford bulls. Therefore, the existence of climatic differences between the temperate and tropical regions evaluated affected Braford bulls' seminal motility and seminal redox homeostasis.

Genetic evaluation of semen production potential in Indigenous bulls using random regression models in a Bayesian framework.

Genetic potential of indigenous bulls with respect to semen production traits over the age of the bulls at semen collection was analyzed using random regression models (RRMs). Data pertaining to 59,641 ejaculates from 189 bulls of 18 indigenous breeds collected from BAIF (Bharatiya Agro Industries Foundation) was utilized for this study. Six semen production traits, viz., ejaculate volume (EV, ml), sperm concentration (SC, 109/ml), initial sperm motility (ISM, %), post-thaw motility (PTM, %), the total number of spermatozoa per ejaculate (TNS, 109/ejaculate), and the theoretical number of semen doses (TNSD) were studied. Univariate and RRM were used to obtain variance components and genetic parameter estimates. Two hundred thousand Gibbs samples were generated for each trait with a burn-in of 20,000 and thinning interval of 50 in a Bayesian framework. Legendre polynomials with orders of fit up to 5 for additive and permanent environmental effects were used. RRM modeled the heritability and repeatability for all ages between 3 and 10 years (productive lifespan). Heritability estimates ranged from 0.18 to 0.36, 0.18 to 0.45, 0.02 to 0.06, 0 to 0.001, 0.09 to 0.32, and 0.14 to 0.42 while the repeatability estimates ranged from 0.41 to 0.72, 0.36 to 0.79, 0.04 to 0.10, 0 to 0.001, 0.37 to 0.56, and 0.32 to 0.57 for EV, SC, ISM, PTM, TNS, and TNSD, respectively. Variability of estimates over the age of the bulls obtained through RRM could be useful to further refine the breeding program for age at selection, deciding the production period and age at culling.

Effects of guanidinoacetic acid and coated folic acid supplementation on growth performance, nutrient digestion and hepatic gene expression in Angus bulls.

To evaluate the impacts of guanidinoacetic acid (GAA) and coated folic acid (CFA) on growth performance, nutrient digestion and hepatic gene expression, fifty-two Angus bulls were assigned to four groups in a 2 × 2 factor experimental design. The CFA of 0 or 6 mg/kg dietary DM folic acid was supplemented in diets with GAA of 0 (GAA-) or 0·6 g/kg DM (GAA+), respectively. Average daily gain (ADG), feed efficiency and hepatic creatine concentration increased with GAA or CFA addition, and the increased magnitude of these parameters was greater for addition of CFA in GAA- diets than in GAA+ diets. Blood creatine concentration increased with GAA or CFA addition, and greater increase was observed when CFA was supplemented in GAA+ diets than in GAA- diets. DM intake was unchanged, but rumen total SCFA concentration and digestibilities of DM, crude protein, neutral-detergent fibre and acid-detergent fibre increased with the addition of GAA or CFA. Acetate:propionate ratio was unaffected by GAA, but increased for CFA addition. Increase in blood concentrations of albumin, total protein and insulin-like growth factor-1 (IGF-1) was observed for GAA or CFA addition. Blood folate concentration was decreased by GAA, but increased with CFA addition. Hepatic expressions of IGF-1, phosphoinositide 3-kinase, protein kinase B, mammalian target of rapamycin and ribosomal protein S6 kinase increased with GAA or CFA addition. Results indicated that the combined supplementation of GAA and CFA could not cause ADG increase more when compared with GAA or CFA addition alone.

Effects of riboflavin supplementation on performance, nutrient digestion, rumen microbiota composition and activities of Holstein bulls.

To investigate the influences of dietary riboflavin (RF) addition on nutrient digestion and rumen fermentation, eight rumen cannulated Holstein bulls were randomly allocated into four treatments in a repeated 4 × 4 Latin square design. Daily addition level of RF for each bull in control, low RF, medium RF and high RF was 0, 300, 600 and 900 mg, respectively. Increasing the addition level of RF, DM intake was not affected, average daily gain tended to be increased linearly and feed conversion ratio decreased linearly. Total tract digestibilities of DM, organic matter, crude protein (CP) and neutral-detergent fibre (NDF) increased linearly. Rumen pH decreased quadratically, and total volatile fatty acids (VFA) increased quadratically. Acetate molar percentage and acetate:propionate ratio increased linearly, but propionate molar percentage and ammonia-N content decreased linearly. Rumen effective degradability of DM increased linearly, NDF increased quadratically but CP was unaltered. Activity of cellulase and populations of total bacteria, protozoa, fungi, dominant cellulolytic bacteria, Prevotella ruminicola and Ruminobacter amylophilus increased linearly. Linear increase was observed for urinary total purine derivatives excretion. The data suggested that dietary RF addition was essential for rumen microbial growth, and no further increase in performance and rumen total VFA concentration was observed when increasing RF level from 600 to 900 mg/d in dairy bulls.

Spermatozoal transcripts associated with oxidative stress and mitochondrial membrane potential differ between high- and low-fertile crossbred bulls.

The presence of various forms of RNAs having roles in fertilisation and early embryonic development is well documented in mammalian spermatozoa. In the present study, using Agilent microarray platform, we compared sperm mRNA expression profiles between high- and low-fertile crossbred bulls with normal semen parameters. Microarray data acquisition and analysis were performed using GeneSpring GX version software, wherein spermatozoa from high-fertile bulls were kept as control while spermatozoa from low-fertile bulls were considered as treatment group. A total of 6,238 transcripts were detected in crossbred bull spermatozoa; 559 transcripts (>1.5-fold) were differentially regulated between high- and low-fertile bulls. Functional annotation has categorised these transcripts into biological process, cellular, and molecular functions. It was observed that transcripts associated with oxidation reduction process (p = .003), mitochondrial membrane potential (p = .03), were significantly down-regulated while transcripts associated with apoptosis (p = .04) were up-regulated in low-fertile spermatozoa. The dysregulated genes were involved in important cellular pathways including oxidative phosphorylation (p = .002), oestrogen signalling (p = .002), Wnt signalling (p = .035), cGMP-PKG signalling (p = .007) and MAPK signalling (p = .032) pathways. Collectively, the present study discovered profound discrepancies in sperm mRNA expression between high- and low-fertile crossbred bulls, with potential possibilities for their use in fertility prediction.

Production of sexed bovine embryos in vitro can be improved by selection of sperm treatment and co-culture system.

The study investigated the effects of sperm sorting, capacitation treatment and co-cultivation on sexed bovine in vitro embryo production. The effect of treatment and co-culture on production of embryos of the preferred sex from unsorted sperm was also studied. Sperm from five breeding bulls was used for fertilization of mature oocytes as follows: Experiment 1, sorted and unsorted sperm (bulls A-E) treated only with heparin in standard co-cultures; Experiment 2, sorted sperm (bulls A-E) treated with heparin-PHE (penicillamine, hypotaurine, and epinephrine) or heparin-caffeine in drop co-cultures; and Experiment 3, unsorted sperm (bull E) treated with either heparin-PHE or heparin-caffeine in both standard and drop co-cultures. In all bulls, treatment with heparin resulted in significantly (p < .05) reduced cleavage and blastocyst rates from sorted sperm, as compared with those from unsorted sperm. In bulls A, B, D and E, treatment of sorted sperm with heparin-PHE in drops significantly increased the blastocyst rate (p < .05). In unsorted sperm of bull E, heparin-PHE treatment in drops resulted in the XX/XY sex ratio inverse to that obtained by heparin-caffeine treatment in standard co-cultures (32.3%/67.7% and 66.7%/33.3%, respectively). In conclusion, the treatment of sorted sperm with heparin-PHE in modified drop co-cultures can be recommended for production of in vitro sexed embryos. The use of unsorted sperm for production of embryos of the preferred sex by selected capacitation treatment and co-culture can be the method of choice in bulls with low IVF yields from sorted sperm.

Neosporosis in bulls: potential for venereal transmission, and effect on semen quality and production.

Neospora caninum caninum is a protozoan parasite that can cause abortions, birth of congenitally infected calves, and reduced milk production in cattle. Neosporosis in cattle can be transmitted through vertical transplacental transmission from dam to calf, or through horizontal transmission between animals. While it is well established that horizontal transmission from dogs (the definitive host) to cattle (an intermediate host) occurs, it is not clear whether horizontal transmission may occur between cows and bulls during mating. Transmission via infected semen of seropositive bulls was suggested when N. caninum DNA was found in bull semen. The primary objective of this article is to critically evaluate the evidence for venereal transmission of N. caninum between cattle, to determine whether this transmission route is likely to occur. A secondary objective is to review and evaluate published evidence for whether neosporosis in bulls may affect semen quality and production. N. caninum DNA can be detected by PCR in occasional batches of semen straws from bulls that are seropositive for N. caninum. Insemination of heifers with semen experimentally infected with 5 × 104 N. caninum tachyzoites has caused persistent seroconversion in some animals. However, the highest concentration of parasites in the semen of naturally infected bulls was estimated, using quantitative PCR, as 10 tachyzoites. Thus it appears that, whether used for natural or artificial insemination, the semen of bulls naturally infected with N. caninum, does not contain the necessary intrauterine dose of tachyzoites to cause persistent seroconversion in a heifer or cow. Thus, based on the available evidence, the venereal transmission of N. caninum via bull semen is extremely unlikely. Nevertheless, neosporosis in bulls may influence semen quality and production. Epididymal semen concentration, viability, and motility appeared "to be" significantly decreased in N. caninum-seropositive bulls compared to seronegative bulls. Furthermore, semen production could be altered if neosporosis affects the thyroid gland. The thyroid hormones such as triiodothyronine and thyroxine (T4) may affect testicular development. Significantly lower concentrations of T4 were observed in the blood of N. caninum-seropositive, compared with those of seronegative bulls. Longitudinal studies are needed to confirm the long-term effect and significance of neosporosis on semen quality and thyroid hormone concentrations in bulls.