Long term effects of immunization against inhibin on fresh and post-thawed semen quality and sperm kinematics during low and peak breeding seasons in Beetal bucks

This study was conducted to evaluate the response of Bali bulls (Bos javanicus) to different semen collection methods and their effects on fresh and post-thawed semen quality. The collection methods employed were electro-ejaculation (EE), transrectal massage (RM) and RM followed by EE (RM+EE). A total of 25 untrained Bali bulls (age between 2 and 4years old) were subjected to the different semen collection methods. Fresh semen samples from all the 25 bulls were evaluated for volume, pH, general motility, live/dead ratio and abnormality using the conventional method. For fresh and frozen samples collected by EE and RM from 10 bulls, computer-assisted semen analysis system was used for precise quantitative measurement of motility, velocity and forward progression. Accucell photometer was used to measure sperm concentration in all samples, regardless fresh and frozen. Semen samples were obtained 100% of the attempts using EE, 84% using RM and 96% using RM+EE. There were no differences among the collection methods for fresh semen quality characteristics, including motility, morphology and viability, but pH and volume were higher for EE than RM and RM+EE. Higher sperm concentration was observed in semen collected by RM than the other two methods. Different age groups (2-3 and >3-4years old) of the bulls did not show significant differences in volume, pH, sperm concentration, percentages in motility, live/dead ratio and normal sperm morphology. The quality of semen for general and progressive motility, VAP, VSL and VCL and acrosomal integrity after thawing was higher for RM than EE. In conclusion, Bali bulls appeared to respond best to EE and the combination of RM+EE than RM, as a method of semen collection, with a shorter time of stimulation required. Differences in age of the Bali bulls did not affect the semen quality.

In the subtropics, bucks show seasonal breeding patterns, and their semen quality decreases during the non-breeding season. Therefore, breeders tend to improve bucks’ semen quality before the breeding season for higher conception rates. In the current study, we hypothesised that simultaneous administration of equine chorionic gonadotrophin (ECG) and melatonin would improve fresh semen quality in bucks before the breeding season. Nine Beetal bucks were randomly assigned (n=3 per treatment) to three treatments: control, melatonin, and melatonin + ECG. Melatonin implants (18 mg; BTC Lab) were placed subcutaneously at the base of the ear. Bucks in the melatonin + ECG treatment were administered ECG (400 IU; Syncro-Part, Ceva Santé Animale) intramuscularly on every fourth day until the end of the experiment. Control bucks were administered normal saline (400 IU; Otuska Pakistan) intramuscularly on every fourth day. Semen was collected twice per week using an artificial vagina (42°C) and immediately evaluated for volume, color, pH, and contaminants. Sperm concentration, motility and kinematics (curvilinear velocity, straight-line velocity, average path velocity, and amplitude of lateral head displacement), viability, DNA, and acrosomal and mitochondrial integrity were monitored using a computer-assisted semen analyzer (AndroVision, Minitube). Weekly concentrations of plasma testosterone and melatonin of all bucks were analysed using radioimmunoassay (Immunotech, Beckman Coulter Ltd.) and enzyme-linked immunosorbent assay (450nm), respectively. Comparisons within and between treatments were made using generalised linear models (repeated-measures analysis of variance). Weekly single-point variance between the treatments was determined (analysis of variance) at P ≤ 0.05 (SPSS ver. 20.0; IBM Corp.). Semen quality (volume, pH, total motility (%), and concentration) improved after Week 4 in the melatonin + ECG treatment compared with the control and melatonin treatments (P<0.05). Similarly, progressive motility (%), viability, DNA, acrosomal and mitochondrial integrity, and sperm kinematics (curvilinear velocity, straight-line velocity, average path velocity, and amplitude of lateral head displacement) improved (P<0.05) after Week 4 in the melatonin + ECG treatment. Similarly, non-viability and ratio of abnormal spermatozoa decreased by Week 3 in the melatonin + ECG treatment (P<0.05) compared with the control and melatonin treatments. Likewise, plasma testosterone concentration (ngmL−1) of bucks was higher (P<0.05) at Week 3 in the melatonin + ECG treatment (4.2±0.2) than in the melatonin (0.8±0.1) and control (1.2±0.1) treatments. Within the melatonin + ECG treatment, plasma testosterone concentration was higher (P<0.05) at Week 5 (4.9±0.2) and Week 9 (4.5±0.1) than at Week 3 (4.2±0.2). Plasma melatonin concentration (pgmL−1) increased (P<0.05) from Week 5 onward in the melatonin + eCG (12.5±0.1) and melatonin (10.2±0.1) treatments compared with the control (2.65±0.1). In conclusion, the simultaneous administration of melatonin and ECG improved fresh semen quality in Beetal bucks.

The aim of this study was to evaluate parameters indicative of sperm quality of fresh and post-thawed semen of Aberdeen Angus, Holstein and Nelore bulls. Thirty-nine bulls were used: Aberdeen Angus (n=13), Holstein (n=13) and Nelore (n=13). The ejaculate collects were performed twice a week using artificial vagina, totaling 792 semen collections, 307 for Aberdeen Angus, 225 for Holstein and 260 for Nelore bulls. After collection, fresh semen was evaluated and semen freezing was performed. After freezing, the batches were thawed and progressive motility was determined. The analysis of fresh semen showed that there was no difference (P = 0.053) between the Aberdeen Angus and Nelore breeds, while ejaculates from Holstein bulls showed a statistical difference (P = 0.024). As well, a difference (P<0.001) was identified in the sperm concentration of the three breeds. In the samples evaluated after thawing, a statistical difference was observed between Holstein and Nelore breeds (P<0.001), while the values of the Angus breed were similar to the other two breeds. The difference in motility of fresh and post-thawing semen showed that Nelore and Angus bulls showed greater variation in values between the analyzes (26.0±8.9% and 25.3±8.4%, respectively) showing a significant difference (P<0.001) in relation to Holstein bulls (20.6±9.3%) that obtained the smallest difference. The analysis of fresh and post-thawing semen did not show any significant difference (P=0.13) between breeds. In conclusion, the semen cryopreservation process causes a decrease in the physical parameters of the semen and these quality losses suffer interference according to the breeds.

The present study was conducted to analyze the fresh and post-thaw semen quality and fertility from native bulls of Red Chittagong Cattle (RCC), BLRI Cattle Breed 1 (BCB1), and Munshiganj Cattle of Bangladesh. One hundred and seventy-two ejaculates were collected by artificial vagina set and semen analysis was performed using Computer Assisted Sperm Analyzer (CASA) at Bangladesh Livestock Research Institute. Commercial extender (AndroMed) was used to dilute the fresh semen. After equilibration (4°C for 4 hr), freezing was done using a programmable bio-freezer. Post-thawed semen was evaluated for sperm motility and kinematics. Cryopreserved semen straws were used for artificial insemination (AI) and determined the bull fertility based on 60 days non-return rate. Motility of the sperm differs significantly (p 0.05) among the bulls but the highest concentration was found in Munshiganj bull (1669.60 ± 192.07 million/ml) followed by RCC (1648.70 ± 91.07 million/ml) and BCB1 bull (1481.60 ± 167.35 million/ml). Moreover, the highest bent tail (5.89 ± 0.75%), coiled tail (1.01% ± 0.22%) and distal mid-piece reflex (2.26% ± 0.28%) were observed in BCB1 followed by Munshiganj and RCC. Amplitude of lateral head displacement (ALH) was recorded higher in post-thaw than in fresh semen. Kinematics parameters of post-thaw semen decreased than fresh semen irrespective of genotypes. More number of doses/ejaculates can be produced from Munshiganj bull (394.34 ± 127.95) followed by RCC (349.01 ± 120.91) and BCB 1 bulls (331 ± 98.99). Fertility rate does not differ among the bulls (p > 0.05) but the highest value was found for RCC (62.06% ± 1.94%) bulls. Therefore, it can be concise that, the quality of Red Chittagong Cattle semen is better than BCB 1 and Munshigang bull.

Quality of Fresh, Cooled, and Frozen Semen From Stallions Supplemented with Antioxidants and Fatty Acids

Dietary n-3 PUFAs improve fresh and post-thaw semen quality in Holstein bulls via alteration of sperm fatty acid composition

Effect of omega-3 and omega-6 polyunsaturated fatty acid enriched diet on plasma IGF-1 and testosterone concentration, puberty and semen quality in male buffalo

The aim of this study was to investigate the relationship between the levels of insulin-like growth factor-1 (IGF-1) in serum and seminal plasma and the characteristics of semen in Beetal bucks (Capra hircus). A total of 12 adult Beetal bucks were involved in the study, with each buck providing six ejaculates collected using a standard artificial vagina (n = 72 total). Only qualified semen samples (volume of 0.7 mL, a mass motility rating of 3+ or higher on a 0-+ scale, and individual progressive motility of 80% or more) divided into three fractions were processed for estimation of IGF-1 and other seminal parameters like motility, viability, acrosome integrity, sperm abnormality and superoxide dismutase (SOD) activity. The first and second fraction were diluted and extended with Optixcell extender (1:15 ratio). The first ejaculate fraction was processed for studying fresh semen parameters and the second fraction was cryopreserved for evaluating frozen semen parameters. French mini straws (0.25 mL) were used for semen filling, and polyvinyl alcohol powder of different colours was used for sealing the extended semen. The third fraction of each ejaculate was centrifuged at room temperature (1100 × g for 7 min) to separate the seminal plasma. Additionally, blood samples were taken from each buck on the same day as semen collection, resulting in a total of 36 blood samples. The results revealed a significant positive correlation (r = .4243; p < .05) between the concentration of IGF-1 in both serum and seminal plasma of the Beetal bucks. Furthermore, the concentration of IGF-1 in serum showed significant positive correlations with sperm viability (r = .554; p < .05), acrosome integrity (r = .527; p < .05), post-thaw sperm motility (r = .407; p < .01), post-thaw sperm viability (r = .426; p < .01) and post-thaw acrosome integrity (r = .333; p < .05). However, it had a significant negative correlation with SOD activity in fresh semen (r = -0.458; p < .01). Moreover, the concentration of IGF-1 in seminal plasma demonstrated significant positive correlations with individual progressive motility (r = .341; p < .05), sperm viability (r = .527; p < .05), acrosome integrity (r = .539; p < .05), sperm plasma membrane integrity (r = .464; p < .05), post-thaw sperm motility (r = .644; p < .01), post-thaw sperm viability (r = .643; p < .01), post-thaw acrosome integrity (r = .487; p < .01) and post-thaw sperm plasma membrane integrity (r = .521; p < .01). Additionally, it showed a significant negative correlation with SOD activity in both fresh semen (r = -0.714; p < .01) and frozen semen (r = -0.558; p < .01) of Beetal bucks. Based on these findings, IGF-1 in seminal plasma can be considered as a potential biomarker for the selection of bucks for breeding purposes.

Effect of feeding a docosahexaenoic acid-enriched nutriceutical on the quality of fresh and frozen-thawed semen in Holstein bulls

Effect of organic selenium dietary supplementation on quality and fertility of cryopreserved chicken sperm

Melatonin along with eCG improves fresh semen quality and plasma concentrations of melatonin and testosterone during non-breeding season in Beetal bucks

Aging roosters typically exhibit subfertility with decreasing semen quality, whereas Thai native roosters reared in rural areas were raised for a longer duration than their usual lifespan. The present study therefore aimed to assess the effect of selenium supplementation as an antioxidative substance in diets to improve the semen cryopreservation of aged roosters. Semen samples were collected from young (n=20) and aged (n=20) Thai native roosters (Pradu Hang Dum) at 36 and 105 weeks of age when starting the experiment, respectively. They were fed diets in non-supplemented or supplemented selenium (0.75 ppm). Fresh semen quality and lipid peroxidation of fresh semen was evaluated before cryopreservation using the traditional liquid nitrogen vapor method. Post-thaw sperm quality and fertility potential were determined. Advancing age is unrelated to decreasing fresh semen quality (p > 0.05). However, lipid peroxidation in rooster semen depended on age, and the malondialdehyde (MDA) concentration increased in aged roosters (p < 0.05). Selenium supplementation in diets significantly decreased the MDA concentration and increased the sperm concentration (p < 0.05). In contrast, cryopreserved semen was affected by advancing rooster age, and selenium influenced sperm quality (p < 0.05). Younger roosters had higher post-thaw sperm quality and fertility potential than aged roosters (p < 0.05). Likewise, diet selenium supplements improved post-thaw sperm quality and fertility compared with the non-supplement group. Rooster's age does not influence the rooster sperm quality of fresh semen, while sperm cryotolerance and fertility were greater in young roosters than in aged roosters. However, those of aged roosters could be improved by dietary selenium supplementation.

The success of artificial insemination (AI) with frozen-thawed semen in cattle is influenced by both female factors and sperm quality. In terms of sperm quality, prior studies indicate that the ability of frozen-thawed bovine sperm to fertilise an oocyte is dependent on their quality and resilience to cryopreservation. Cryopreservation induces oxidative stress, leading to ultrastructural damage in the sperm. This study aimed to determine whether the quality of fresh semen can identify bulls with good and poor sperm freezability. This difference between fresh and frozen semen from the same bull allows us to predict fertility. Motility and kinetic parameters were assessed using computer-assisted sperm analysis (CASA), while six functional variables were evaluated through flow cytometry, both before and after the freeze-thaw process on the sperm from 13 bulls. Invivo fertility was measured using 90-day non-return rates. The principal component analysis (PCA) of eight sperm variables post-thaw identified one principal component explaining 81.19% of the total variance and classified the bulls into two groups: Poor freezability bulls (progressive motility: 48.12% ± 8.41%; viability: 77.51% ± 7.61%) and good freezability bulls (progressive motility: 58.64% ± 6.64%; viability: 88.12% ± 2.52%). Bulls with higher freezability showed better sperm viability and motility, as well as lower levels of ROS, superoxides and intracellular calcium before cryopreservation that were significantly correlated with higher non-return rates (NRR). The results underscore the importance of assessing the quality and functionality of fresh semen to predict the fertility potential of cryopreserved sperm. This approach can aid in selecting ejaculates with the best potential for successful artificial insemination, ultimately improving reproductive performance in dairy cattle.

Buffalo bull spermatozoa are rich in polyunsaturated fatty acids and are prone to lipid peroxidation. We hypothesised that lipid peroxidation in buffalo bull semen will increase with age and will affect the semen quality. The objective was to compare malondialdehyde (MDA) concentration and the quality of fresh and frozen-thawed semen between aged (13.6 ± 1.0 years; n = 3) and young (3.4 ± 0.3 years; n = 3) Nili-Ravi bulls. The concentration of MDA did not differ (p >.05) between aged vs. young bulls in fresh (2.3 ± 0.2 vs. 2.9 ± 0.7 nmol mL−1), frozen-thawed (53.1 ± 2.8 vs. 48.4 ± 2.6 nmol mL−1) semen and seminal plasma (5.71 ± 0.97 vs. 5.19 ± 1.36 nmol mL−1), respectively. In fresh semen, sperm motility and total concentration did not differ (p > .05) between aged and young bulls. The volume of fresh semen increased (p > .05) while sperm viability and DNA integrity decreased (p < .05) in aged vs. young bulls. In frozen-thawed semen, sperm motility, viability and DNA integrity decreased (p < .05) in aged vs. young bulls. In frozen-thawed vs. fresh semen, MDA level increased within young (48.4 ± 2.6 vs. 2.3 ± 0.2 nmol mL−1) and aged bulls (53.1 ± 2.8 vs. 2.9 ± 0.7 nmol mL−1). Conversely, sperm motility and viability decreased (p < .05) within the age groups between fresh and frozen-thawed semen. In conclusion, (1) lipid peroxidation (MDA) does not increase due to age; however, it is negatively associated with semen quality and (2) cryopreservation steps up lipid peroxidation irrespective of age and deteriorates semen quality of Nili-Ravi bulls.

The libido of buffalo bulls is crucial factor for influencing the semen production for artificial insemination programs. The present study investigated the relationship of libido with hormonal profiles and semen quality of Murrah (n = 5) and Nili Ravi (n =1) buffalo bulls classified into good (n = 2), average (n = 2) and poor (n = 2) libido groups based on reac tion time (&lt; 3 min, between 3-6 min and &gt; 6 min, respectively). Serum testosterone, estradiol, prolactin and kisspeptin levels were assayed post-GnRH administration using ELISA kits, while serum nitric oxide (NO) was assayed using Griess reaction kit (ELISA kit). Fresh semen quality (volume, concentration, total motility) and post-thaw frozen semen qual ity, including CASA-derived motion traits and morphological parameters, were evaluated. Serum testosterone, estradiol, prolactin kisspeptin and NO didn’t significantly differ across libido groups, however testosterone to estradiol (T/E) ratio was significantly higher in bulls exhibiting good libido traits (p &lt; 0.05). Among hormones, solely kisspeptin and T/E ratio showed significant positive correlation with each other. Fresh semen parameters (volume, sperm concentration and fresh motility) were significantly superior in the good libido buffalo bulls (p &lt; 0.01). For post-thaw semen, motility parameters were similar across groups, while several kinetic motion traits (DCL, DSL, DAP, VCL, VSL, VAP, radius, ALH, HAC) were significantly higher in good libido group (p &lt; 0.01). Sperm viability, acrosomal integrity, and mito chondrial membrane potential didn’t differ significantly (p &gt; 0.05). Evaluating correlation among serum hormones and semen quality, T/E ratio showed significant (p &lt; 0.05) correlation with DCL, DSL, DAP and VSL, while relation among other parameters remained non-significant (p &gt; 0.05). In conclusion, good libido of buffalo bulls positively influences their semen quality. Among serum hormones, T/E ratio influences the libido exhibition in buffalo bulls