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This study aimed to examine how varying concentrations (0.5%, 1.0%, 2.0%, 3.0%, and 4.0%) of nano-L-a phosphatidylcholine (nano-L-a-PC) affected the quality of post-thawed buffalo bull semen compared to the same concentrations of L-a-phosphatidylcholine (L-a-PC), while 20% egg yolk (EY) was used as a control. Ejaculates (n = 72) were collected from fertile buffalo bulls (n = 12) for 6 weeks. The ejaculates were pooled and divided into 11 groups (3 aliquots per group) extended with 20% EY (Group 1 without supplement), 0.5%, 1.0%, 2.0%, 3.0%, and 4.0% nano-L-a-PC (Groups 2-6), and 0.5%, 1.0%, 2.0%, 3.0%, and 4.0% L-a-PC (Groups 7-11) in tris buffer. After cryopreservation procedures, semen samples were thawed at 37 °C and then evaluated for sperm kinematics, acrosomes, plasma membranes, and DNA integrities. The seminal plasma was analyzed for malondialdehyde (MDA), superoxide dismutase (SOD), glutathione reduced (GSH), and catalase (CAT). Results demonstrated that extended semen samples containing 1.0-3.0% nano-L-a-PC and 1.0-4.0% L-a-PC showed high total motility compared to control, whereas at 0.5-4.0% of nano-L-a-PC and L-a-PC showed high progressive motility. Significant (P< 0.05) wobble levels were seen at 0.5, 1.0, and 3.0% nano-L-a-PC, and distance average path and velocity average path at 2.0% L-a-PC. Acrosome and plasma membrane integrities were markedly elevated (P< 0.0001) at 2.0% nano-L-a-PC and 4.0% L-a-PC. High DNA integrity metrics were noticed at 0.5-1.0% nano-L-a-PC, as well as 3.0% and 4% L-a-PC. Low MDA and high (SOD, GSH, and CAT) levels were observed at 2.0% nano-L-a-PC and 4.0% L-a-PC. In conclusion, the optimum concentration of nano-L-a-PC that improved semen quality was 2.0%, which is roughly equivalent to the effect of 4.0% L-a-PC. [ABSTRACT FROM AUTHOR]

Simple Summary: Cryopreserved semen, widely utilized in the artificial insemination of domestic animals, encounters challenges due to oxidative damage. This study investigated the effects of the antioxidant PQQ on the semen quality of Holstein bulls during cryopreservation and elucidated the associated mechanisms. Our findings revealed that PQQ significantly enhanced sperm motility, membrane integrity, acrosome integrity, and ATP levels, while simultaneously reducing MDA and ROS levels. Additionally, increased PGAM2, CAPZB, CAT, SOD1, and GPX1 protein expressions were also observed in the PQQ-treated group. Based on these results, PQQ not only improved semen quality but also mitigated oxidative stress, thereby enhancing the efficacy of sperm cryopreservation in bulls. Cryopreserved semen is extensively utilized in the artificial insemination (AI) of domestic animals; however, suboptimal conception rates due to oxidative damage following AI continue to pose a challenge. The present study investigated the effects of Pyrroroquinoline Quinone (PQQ), a novel antioxidant, on the semen quality of Holstein bulls during cryopreservation, as well as its potential molecular mechanisms. Semen samples were diluted with varying concentrations of PQQ (0, 50 μmol/L, 100 μmol/L, 150 μmol/L) prior to cryopreservation. Following the freeze–thaw process, a comprehensive evaluation was conducted to assess sperm motility, plasma membrane integrity, acrosome integrity, and the levels of reactive oxygen species (ROS), malondialdehyde (MDA), and adenosine triphosphate (ATP). Western blot analysis was employed to examine the levels of proteins including PGAM2, CAPZB, CAT, SOD1, and GPX1. Notably, the inclusion of 100 μmol/L PQQ significantly enhanced sperm motility, membrane integrity, and acrosome integrity post freeze–thawing (p < 0.05). Furthermore, the group treated with 100 μmol/L PQQ exhibited reduced levels of MDA and ROS (p < 0.05), while ATP levels were significantly elevated (p < 0.05). Interestingly, treatment with 100 μmol/L PQQ resulted in decreased consumption of PGAM2, CAPZB, CAT, SOD1, and GPX1 proteins in sperm after freeze–thawing, compared to the control group (p < 0.05). These findings indicate that PQQ treatment enhances the quality of bull semen, mitigates oxidative stress damage, and ultimately improves the efficacy of sperm cryopreservation. [ABSTRACT FROM AUTHOR]

This study aims to compare the efficacy of computer‐assisted sperm analysis (CASA) and smartphone‐applied sperm analysis (SASA) in assessing the quality of frozen–thawed bull semen. A total of 75 straws (n = 75) semen samples were used from different production batches of five Holstein bulls. The semen analyses were conducted in three groups: Group I (CASA‐37°C), semen samples were evaluated using the CASA system at 37°C (n = 25); Group II (SASA‐25°C), semen samples were assessed using the SASA system at a temperature of room heat (25°C) (n = 25); and Group III (SASA‐37°C), semen samples were evaluated using the SASA system at 37°C (n = 25). The frozen–thawed bull semen samples were analysed in terms of total motility (TM), progressive motility (PM), immotile, velocity average path (VAP), velocity curve linear (VCL), velocity straight line (VSL) and sperm concentration. There was no significant difference between the groups in terms of spermatozoa concentration (p >.05). However, significant differences among the groups were observed for total motile spermatozoa values (p <.001). Values of progressive motile spermatozoa were lower in Group I and Group II compared to Group III (p <.001). The immotile spermatozoa values were significant between the groups (p <.001) and were found to be proportional to total motile spermatozoa values. Additionally, the VAP, VCL and VSL values were comparable between Group II and Group III, but lower when compared to Group I. In conclusion, the results of the study demonstrate that the Sperm Cell™ system can accurately analyse the concentration of frozen–thawed bull semen. The analyses performed at room temperature indicate a parallelism between the PM value and CASA results. However, it is thought that SASA devices require a series of standardization studies in different semen extenders, different sample concentrations and different animal species, analogous to the standardization evolution process of CASA devices in semen analysis. [ABSTRACT FROM AUTHOR]

Regular verification of the quality of cryopreserved semen derived from native cattle is one of the tasks performed at the bank as recommended by the Food and Agriculture Organization. The purpose of the present study was to evaluate the quality of semen from PR bulls stored for 40, 50 and 60 years in the BMB using standard evaluation parameters such as sperm motility as well as structural-functional parameters such as plasma membrane integrity, transmembrane mitochondrial potential and sperm chromatin damage. Semen pellets from 27 PR bulls (3 ejaculates/bull) were tested. The data were analysed by one-way and two-way ANOVA, and the significance of the difference (P≤ 0.01) between the means was determined using Duncan's test. Our study results revealed that the long-term storage of semen had no effect on sperm characteristics after thawing. However, statistically significant differences (P≤0.01) in sperm plasma membrane integrity and transmembrane mitochondrial potential, following storage in liquid nitrogen were noted between bulls at all time points. However, there were no significant differences (P>0.01) in sperm chromatin damage between breeds or between different storage times, and the degree of DNA fragmentation ranged from 0.4 to 0.8%. [ABSTRACT FROM AUTHOR]

The present study was designed to determine the effect of different concentrations of quail egg yolk (5%, 10%, 15% and 20%) in tris based semen extender on progressive motility of Sahiwal bull spermatozoa at pre-freeze as well as post-thaw stage. The progressive motility significantly (P<0.01) improved in 10% and 15% quail egg yolk concentration at prefreeze stage whereas it increases significantly in 10% quail egg yolk concentration at post-thaw stage. The findings of the present study suggested that quail egg yolk improves post-thaw sperm progressive motility at 10% concentration. [ABSTRACT FROM AUTHOR]

The present investigation was undertaken on cattle bull semen reared at Frozen Semen Bull Station, Karimnagar, Telangana. The study was carried out on four cattle breeds (3 to 5 years of age) viz., Gir, Ongole, Jersey and Crossbred Jersey (CBJY). Semen was collected twice weekly from each bull using artificial vagina technique with a total of 32 ejaculates evaluated under study. The semen was diluted with Egg Yolk Tris Citrate (EYTC) extender and was evaluated for physical characteristics of neat semen, motility parameters and sperm viability percentage. The investigation revealed that Gir bull semen had significantly higher (p<0.05) ejaculate volume, mass motility, individual motility, sperm concentration compared to other breeds in present study. Whereas, Jersey bull semen had significantly higher (p<0.05) sperm viability percentage. [ABSTRACT FROM AUTHOR]

The purpose of this study was to improve the quality of frozen--thawed Piedmontese bull semen by incorporating MitoTEMPO (MT) in extended semen before cryopreservation. Semen was collected from 4 fertile bulls, using an artificial vagina, once weekly for 6 consecutive weeks. Semen samples were pooled, diluted with Bullxcell® extender, and supplemented with different concentrations of MT (0 as control, 5, 10, 20, 40, and 80 µM) before cooling, equilibration, and freezing procedures. The frozen--thawed semen was assessed for motility, vitality, acrosome intactness, plasma membrane integrity, DNA integrity, apoptosis, mitochondrial membrane potential, intracellular ROS level and in vitro fertilizing capability. The results showed that MT at concentrations of 10, 20, and 40 µM improved the total, progressive, and rapid motility directly after thawing while, at the highest tested concentration (80 μM), it decreased the progressive and rapid motility after 1, 2, and 3 h of incubation. The sperm kinetics including STR and LIN were noticeably increased at concentrations of 10, 20, and 40 µM directly after thawing (0 h), whereas the MT effect was variable on the other sperm kinetics during the different incubation periods. MitoTEMPO improved the sperm vitality at all tested concentrations, while the acrosomal and DNA integrity were improved at 20 µM and the mitochondrial membrane potentials was increased at 80 µM. The cleavage and blastocyst formation rates were significantly increased by using semen treated with 20 µM MT compared with controls. These findings suggest a potential use of MT mainly at a concentration of 20 µM as an additive in the cryopreservation media of bull semen to improve sperm quality. [ABSTRACT FROM AUTHOR]

The goal of current investigation was to ascertain the impact of carnitine as an additive in tris-egg yolk-based extender on Hariana bull semen (32 ejaculates) chosen for cryopreservation. Progressive motility, viability, HOS response, and acrosomal integrity were estimated after equilibration (pre-freeze) and after thawing. The activity of enzymes malondialdehyde (MDA), glutathione-S-transferase (GST), and superoxide dismutase (SOD) were quantified in post-thaw seminal plasma. Semen ejaculates from Hariana bulls (n=4) were diluted in tris egg yolk glycerol (TEYG) and were divided into four equal groups. Aliquot/Group I was processed without carnitine and served as control, while Groups II, III and IV received carnitine at concentration of 2.5 mM, 5 mM and 10 mM per 80x106 spermatozoa, respectively. Carnitine at 5 mM concentration significantly (p<0.05) enhanced sperm viability, acrosomal integrity, and intact plasma membranes both at pre-freeze and post-thaw stage. The addition of 5 mM and 2.5 mM carnitine reduced oxidative stress considerably (p<0.05). The results of this investigation revealed that carnitine has cryoprotective properties on bull spermatozoa via reducing oxidative stress and free radicals. Hence to improve post-thaw sperm functional dynamics, it was suggested that 5 mM of carnitine be supplemented as an additive to semen extender. [ABSTRACT FROM AUTHOR]

Semen samples contain bacteria originating from the animal urogenital tract, environment, and/or contamination during semen processing, negatively affecting sperm quality by producing toxins and/or competing for nutrients in extenders. The aims of this study were to evaluate two methods of Single-layer centrifuges (SLC), high and low density colloid, as a method for bacterial removal from bull semen, and to evaluate sperm quality after treatment. In total, semen samples from 20 bulls (3 ejaculates per bull) were used in this study. Bacterial reduction was evaluated by bacterial quantification (colony forming unit - CFU/mL) while bacterial identification was performed by matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) after culturing bacteria on blood agar. Sperm motility parameters were evaluated by Computer Assisted Sperm Analyses (CASA), and sperm chromatin structure assay (SCSA) by Flow cytometry. Both, High and Low density SLC reduced number of bacteria significantly (p < 0.001) compared with control. The difference in bacterial count between High and Low SLC was also significant (p < 0.001). Furthermore, High density SLC was successful in removing almost all Bacillus and Proteus spp. Most CASA parameters were significantly improved after both treatments (p < 0.001, p < 0.01, p < 0.05). The Deoxyribonucleic acid (DNA) fragmentation index evaluated by SCSA in High (p < 0.01) and Low (p < 0.05) SLC group differed significantly compared with control. Single-layer centrifugation (SLC) with either a high or a low density colloid is a suitable method for bacterial removal in bull semen. [ABSTRACT FROM AUTHOR]

Simple Summary: As the basis of artificial insemination technology, semen cryopreservation technology is a very valuable technology in the cattle industry and directly affects the success rate of fertilization. The effectiveness of semen cryopreservation is closely associated with quality alterations of fresh semen and semen diluent during semen cryopreservation. Therefore, in this study, in order to improve the effect of semen cryopreservation and fresh semen quality of breeding bulls, enough antioxidant-efficient grape seed proanthocyanidins were added to semen diluent and breeder bull diets. It was found that grape seed proanthocyanidins as a semen diluent extender could improve frozen–thawed sperm morphology and quality (including frozen–thawed sperm motility, membrane integrity, acrosome integrity, and mitochondrial activity). As a dietary additive, it could improve sperm motility, reduce sperm deformity rate, and improve the quality of fresh semen. This study revealed that grape seed proanthocyanidins could improve effect of semen cryopreservation and the reproductive performance of breeding bulls, which was significantly related with the antioxidant properties of grape seed proanthocyanidins. This research promotes the preservation and breeding of bovine semen and provides a basis for the fundamental science of mammalian gametes and reproductive biotechnology. Oxidative stress leads to a decrease in semen quality during semen cryopreservation and fresh semen production. Grape seed proanthocyanidins (GSPs) are endowed with well-recognized antioxidant, anti-inflammatory, anti-cancer, and anti-aging activities. Therefore, the objective of this experiment was to explore the effects of GSPs on the quality of fresh and cryopreserved semen to provide a basis for GSPs as a new dietary additive and semen diluent additive for males' reproduction. Fresh semen from three healthy bulls aged 3 to 5 years old were gathered and mixed with semen diluents dissolved with 0 µg/mL, 30 µg/mL, 40 µg/mL, 50 µg/mL, and 60 µg/mL GSPs respectively. The motility, physiological structures (acrosome integrity, membrane integrity, mitochondrial activity), and antioxidant capacity of frozen–thawed sperm were measured after storage in liquid nitrogen for 7 days (d). Bulls were fed with 20 mg/kg body weight (BW) GSPs in their diet for 60 days; the weight of the bull is about 600 kg. Then, the reproductive performance and antioxidant indexes of bulls were measured before and after feeding. The results demonstrated that GSPs supplementation significantly increased sperm motility, physiological structures, GSH-Px, and CAT enzyme activities and significantly decreased MDA content in sperm during semen cryopreservation. The optimal concentration of GSPs was 40 µg/mL (p < 0.05). After 20 mg/kg (body weight) GSP supplementation, sperm motility was significantly heightened (p < 0.05), the sperm deformity rate was significantly reduced (p < 0.05), and antioxidant enzyme activities (such as SOD, CAT, and GSH-Px) were significantly enhanced (p < 0.05), and the production of MDA was significantly suppressed (p < 0.05) in serum compared with that before feeding. In conclusion, these results reveal that a certain concentration of GSPs has a good protective effect on sperm damage caused by semen cryopreservation and the reproductive performance reduction caused by stress in bulls, which may be attributed to the antioxidant function of GSPs. In summary, GSPs are a useful cryoprotective adjuvant and dietary additive for bull sperm quality. [ABSTRACT FROM AUTHOR]

The aim of the present study was to evaluate the effects of insulin supplemented extender on bull semen at a post-thaw stage. Semen samples were collected four times from a Zavot bull which is endangered species in Türkiye. Semen samples were diluted with Tris based extender supplemented with 10 IU or 15 IU of insulin and control. Motility and plasma membrane integrity were evaluated by a phase-contrast microscope. Acrosome integrity (Fitc-Peanut Agglutinin), mitochondrial membrane potential (Rhodamine 123), nitric oxide level (4,5-Diaminofluorescein-2/diacetate) were evaluated with dual staining propidium iodide using flow cytometry. Motility and plasma membrane functional integrity were better preserved compared to control group (P<0.05). Acrosome integrity results were statistically similar between control and 10 IU insulin groups (P>0.05), but increased insulin negatively affected acrosome integrity (p<0.05). Mitochondrial membrane potential was found to be higher when compared all cells in insulin containing groups compared to control (P<0.05). However, results were found similar in all groups with PI negative/mitochondrial membrane potential positive (P>0.05). There were not any significant differences among groups in terms of nitric oxide level (P>0.05). In conclusion, it was thought that insulin involved in energy metabolism and improved preservation of bull semen. [ABSTRACT FROM AUTHOR]

Sperm survives for a very short time in fresh semen, and slow cooling to 5.00 °C kills a large number of sperms. This study was aimed to compare the semen quality parameters and anti-oxidant levels in four extenders (manual, Triladyl, Steridyl and AndroMed). Semen samples were obtained from a total number of 12 dual-purpose Simmental bulls kept in the Simmental Cattle Breeding Center for a period of 3 months using an artificial vagina. Sperm viability, motility, abnormal morphology, plasma membrane integrity, DNA damage, chromatin quality, total antioxidant capacity (TAC) and lipid peroxidation were evaluated. The highest progressive motility, viability, plasma membrane integrity, and TAC and the lowest levels of malondi-aldehyde in the frozen-thawed semen belonged to the semen group frozen with Triladyl. Parameters of motility were higher in the frozen-thawed semen with Triladyl than in other groups, indicating a significant difference from the manual extender. Among the extenders studied, Triladyl was the most suitable for semen freezing in Simmental bulls. [ABSTRACT FROM AUTHOR]

Introduction: Flaxseed oil (FO) and vitamin E (VE) both have antioxidant effects on sperm. The present study investigated the effects of dietary supplementation with FO and/or VE on semen quality. Methods: 16 fertile Simmental bulls were selected and randomly divided into 4 groups (n = 4): the control group (control diet), FO group (control diet containing 24 g/kg FO), VE group (control diet containing 150 mg/kg VE) and FOVE group (control diet containing 150 mg/kg VE and 24 g/kg FO), and the trial lasted 10 weeks. Results: The results showed that the addition of FO independently can increase sperm motion parameters, the levels of catalase (CAT), glutathione peroxidase (GSH-Px), testosterone (T) and estradiol (E2), while reduce oxidative stress in seminal plasma (P < 0.05). Supplement of VE independently can increased the motility, motility parameters, CAT and superoxide dismutase (SOD) levels, and reduce oxidative stress in seminal plasma (P < 0.05). There was an interaction effect of FO × VE on motility and reactive oxygen species (ROS), while GSH-Px and ROS were affected by week × VE 2-way interaction, levels of T and E2 were also affected by the dietary FO × week interaction (P < 0.05). The triple interaction effects of FO, VE and week were significant for malondialdehyde (MDA) (P < 0.05). Compared with the control group, sperm from the FOVE group had a significantly higher in vitro fertilization (IVF) rate, and subsequent embryos had increased developmental ability with reduced ROS levels at the eight-cell stage, then increased adenosine triphosphate (ATP) content and gene expression levels of CAT, CDX2, Nanog, and SOD at the blastocyst stage (P < 0.05). Metabolomic and transcriptomic results indicated that dietary supplementation of FO and VE increased the expression of the metabolite aconitic acid, as well as the expression of ABAT and AHDHA genes. Conclusion: With in-silico analysis, it can be concluded that the effects of dietary FO and VE on improving semen quality and embryo development may be related to increased aconitic acid via the ABAT and AHDHA genes involved in the propionic acid metabolism pathway. [ABSTRACT FROM AUTHOR]

The rapid emergence of antibacterial resistance requires alternatives to antibiotics to be found, including for semen preservation. One of the possible alternatives would be to use plant-based substances with known antimicrobial effects. The objective of this study was to test the antimicrobial effect of pomegranate powder, ginger, and curcumin extract in two concentrations on bull semen microbiota after exposure for <2 h and 24 h. An additional aim was to evaluate the effect of these substances on sperm quality parameters. The bacterial count in semen was low from the beginning; however, a reduction was present for all tested substances compared with control. A reduction in bacterial count in control samples was also observed with time. Curcumin at a concentration of 5%, reduced bacterial count by 32% and was the only substance that had a slight positive effect on sperm kinematics. The other substances were associated with a decline in sperm kinematics and viability. Neither concentration of curcumin had a deleterious effect on sperm viability parameters measured by flow cytometry. The results of this study indicate that curcumin extract at a concentration of 5% can reduce the bacterial count and does not have a negative influence on bull sperm quality. [ABSTRACT FROM AUTHOR]

Simple Summary: We evaluated the impact of imported semen of Holstein bulls from different countries on their daughters' economic traits using the Lifetime Net Income (LNI) index in different climates of Iran. The first lactation records were collected from 10 big dairy farms in various regions of Iran. Studied traits included milk, fat and protein yields, calving age and calving interval. There were significant differences between sire groups for studied traits in each climate (p < 0.01). In semi-cold, moderate and warm climates, daughters of French sires had the highest least-square means of LNI index. In cold climate, daughters of German sires had the highest least-square means. The aim of the present study was to evaluate the effects of imported semen of Holstein bulls from different countries on the economic traits of their daughters using the Lifetime Net Income (LNI) index in various climates of Iran. The data included the first lactation records of 274,057 Holstein cows collected during 1993 to 2017 by the Animal Breeding Center of Iran from 10 large dairy farms located in various provinces of Iran. The investigated traits included milk, fat and protein yields, calving age and calving interval. Breeding values of progenies were predicted by the Best Linear Unbiased Prediction (BLUP) method under the multi-trait animal model using DMU software. The genetic-economic merit of the progenies was estimated by the LNI index. There were significant differences between the estimated breeding values (EBVs) of sire groups (based on bull semen origin) for milk, fat and protein yields, calving age and calving interval in each climate (p < 0.01). The obtained results showed that the highest least-square means of LNI index in semi-cold, moderate and warm climates belonged to the daughters of French sires; however, daughters of German sires were estimated to have the highest least-square means in the cold climate. [ABSTRACT FROM AUTHOR]

Schmallenberg orthobunyavirus (SBV) was initially detected in 2011 in Germany from dairy cattle with fever and decreased milk yield. The virus infection is now established in many parts of the world with recurrent epidemics. SBV is transmitted through midges and transplacental. No direct virus transmission including via breeding has ever been demonstrated. In some bulls, however, the virus is detectable transiently, in low to minute quantities, in semen post‐infection. While the infection is considered of low impact for the dairy industry, some SBV‐free countries have adopted a zero‐risk approach requiring bull semen batches to be tested for SBV RNA residues prior to import. This, in turn, obligates a protocol to enable sensitive detection of SBV RNA in semen samples for export purposes. Here, we describe how we established a now ISO/IEC 17025 accredited protocol that can effectively detect minute quantities of SBV RNA in semen and also its application to monitor bull semen during two outbreaks in the United Kingdom in 2012 and 2016. The data demonstrate that only a small number of bulls temporarily shed low amounts of SBV. [ABSTRACT FROM AUTHOR]

Distinguishing between bull Y- and X-bearing sperm populations is advantageous for techniques using sexed bull semen. The aim of this study was to produce a single-chain fragment variable (scFv) antibody against plasma membrane epitopes on bull Y-bearing sperm. Variable heavy (VH)- and variable light (VL)-region genes generated from a hybridoma cell secreting a specific Y-bearing sperm monoclonal antibody (mAb-1F9) were cloned and expressed. The expected sizes of the DNA bands were ∼350 bp for the VH gene and ∼318 bp for the VL gene. The VH and VL genes were generated and used to construct an scFv gene (∼650 bp), which was expressed in E.coli TG1 cells and produced the corresponding soluble scFv antibody. Compared with the parent mAb-1F9, the scFv antibodies presented a high affinity for Y-bearing sperm and low cross-reactivity with X-bearing sperm. An immunofluorescence analysis confirmed that the scFv antibodies and mAb-1F9 recognize epitopes on the Y-bearing sperm surface. The fluorescence signal was strong on the plasma membrane of Y-bearing sperm but very weak for X-bearing sperm. This study aids the application and production of engineered scFv antibodies specific to Y-bearing sperm to distinguish between Y- and X-bearing sperm populations for techniques involving sexed bull semen. [ABSTRACT FROM AUTHOR]

Simple Summary: Female calves are favored for milk production and genetic advancement in the dairy industry, and sex selection by using sexed semen has been long considered. A potential alternative sperm sexing technique is magnetic-activated cell sorting combined with an immunological method that uses scFv antibodies against male-specific sites on Y-chromosome-bearing sperm; however, the technique should be evaluated for validity and accuracy. This study focuses on how well bovine sperm are separated by the use of magnetic microbeads coupled with scFv antibodies against Y-chromosome-bearing sperm (PY-microbeads). The results showed that sexed bovine sperm using PY-microbeads was a highly effective technique for distinguishing X- and Y-chromosome-bearing sperm. It had no negative impact on the quality of X-chromosome-bearing sperm. The technique produced 82.65% of X-chromosome sperm in the X-enriched fraction semen and 81.43% of Y-chromosome sperm in the Y-enriched fraction semen, which was utilized to generate target sexed bovine semen. Sperm sexing technique is favored in the dairy industry. This research focuses on the efficiency of bovine sperm sexing using magnetic-activated cell sorting (MACS) by scFv antibody against Y-chromosome-bearing sperm (Y-scFv) coupled to magnetic microbeads and its effects on kinematic variables, sperm quality, and X/Y-sperm ratio. In this study, the optimal concentration of Y-scFv antibody coupling to the surface of magnetic microbeads was 2–4 mg/mL. PY-microbeads revealed significantly enriched Y-chromosome-bearing sperm (Y-sperm) in the eluted fraction (78.01–81.43%) and X-chromosome-bearing sperm (X-sperm) in the supernatant fraction (79.04–82.65%). The quality of frozen–thawed sexed sperm was analyzed by CASA and imaging flow cytometer, which showed that PY-microbeads did not have a negative effect on X-sperm motility, viability, or acrosome integrity. However, sexed Y-sperm had significantly decreased motility and viability. The X/Y-sperm ratio was determined using an imaging flow cytometer and real-time PCR. PY-microbeads produced sperm with up to 82.65% X-sperm in the X-enriched fraction and up to 81.43% Y-sperm in the Y-enriched fraction. Bovine sperm sexing by PY-microbeads showed high efficiency in separating Y-sperm from X-sperm and acceptable sperm quality. This initial technique is feasible for bovine sperm sexing, which increases the number of heifers in dairy herds while lowering production expenses. [ABSTRACT FROM AUTHOR]

To determine the effects of epigallocatechin‐3‐gallate (EGCG) on the cryopreservation of bovine semen, epigallocatechin‐3‐gallate dissolved with double distilled water to 0.2, 0.4, 0.6 and 0.8 mg/ml were added to the cryopreservation diluent of the bull semen. Then, we used computer‐assisted analysis of semen kinematic parameters, staining method to detect membrane function, acrosome integrity, enzyme‐linked immunosorbent assay to detect catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH‐Px), malondialdehydes (MDA) and reactive oxygen levels. The results showed that adding 0.6 mg/L of epigallocatechin‐3‐gallate could improve the cryopreserved sperm quality, which significantly increased the total motility, distance average path, distance straight line, distance curved line, average path velocity, curvilinear‐velocity, straight‐line velocity, amplitude of lateral head displacement and beat/cross frequency, as well as sperm CAT, GSH‐Px and SOD levels (p < 0.05), whilst reducing the reactive oxygen species and MDA levels (p < 0.05). Hence, these results indicate that the addition of 0.6 mg/ml of EGCG has a protective effect on the cryopreservation of the bovine semen. [ABSTRACT FROM AUTHOR]

In this study, the cryoprotective potential of natural antioxidant curcumin in Hariana bull semen was evaluated as an additive in a tris‐based extender with the assessment of motility and motion parameters of spermatozoa, membrane intactness, progesterone‐receptor binding, protein carbonyl content, cervical mucus penetration, cryocapacitation‐associated and apoptotic‐like changes. The collected ejaculates were divided into five groups in the tris‐based extender (control without curcumin‐I, 10 µM‐II, 25 µM‐III, 50 µM‐IV and 75µM‐V) and were cryopreserved. Groups II and III containing 10 and 25 µM curcumin substantially (p <.05) improved the post‐thaw sperm parameters like viability, motility, and velocity parameters; intact acrosome and membrane; lowered protein carbonyl content; DNA fragmentation and cryocapacitation‐associated changes in comparison to control. It was interesting to note that early apoptotic‐like changes in sperm cells were significantly (p <.05) decreased in Group II along with an increase in a higher population of sperm cells having high mitochondrial transmembrane potential. Higher progesterone‐receptor binding, Vanguard distance and in vitro capacitation response were observed only in Group II (10µM) compared to other groups. In conclusion, curcumin in a semen extender manifests cryoprotective effects and may be incorporated at 10 µM concentration in a Hariana bull semen extender for better post‐thaw sperm quality. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate whether vaccenic acid (VA) added to TRIS semen extender has an effect on freezing bull semen. The semen was collected at intervals of three days through an artificial vagina then pooled. The pooled ejaculates were divided into five components as the control (C; no supplement) and VA (25, 50, 100, and 200 µg / mL) groups then extended with the TRIS-based medium. The samples were left in a cold air cabinet for cooling process (4 °C), then they were equilibrated for 4 h. Afterward, the samples in the groups were frozen by using a programmable freezer. The greatest progressive and total motility results were obtained in the VA25 group (p < 0.001). Besides, the total abnormality and chromatin damage was determined to be on the lowest level in the VA25 group (p < 0.05). Except for VA200, the glutathione level was greater in the treatment groups in comparison to the control group (p < 0.05), while the lowest malondialdehyde was determined in VA25. In conclusion, VA supplemented with TRIS-based extender at the concentration of 25 μg / mL improves the progressive and total motility properties of spermatozoa, reduces the abnormal spermatozoa rate, and has a positive effect on chromatin integrity. [ABSTRACT FROM AUTHOR]

Reports on the use of 16S sequencing for the identification of bacteria in healthy animals are lacking. Bacterial contamination of bull semen can have a negative effect on the sperm quality. The aims of this study were threefold: to identify bacteria in the semen of healthy bulls using 16S sequencing; to investigate the differences in the bacterial community between individual bulls; and to establish if there was a relationship between the bacteria isolated and bull fertility. Semen from 18 bulls of known fertility was used for the DNA extraction and 16S sequencing; 107 bacterial genera were identified. The differences in the amplicon sequence variants (ASVs) and the numbers of genera between bulls were noted. Negative correlations (p < 0.05) between several bacterial genera with Curvibacter, Rikenellaceae RC9-gut-group and Dyella spp. were seen. Other negatively correlated bacteria were Cutibacterium, Ruminococcaceae UCG-005, Ruminococcaceae UCG-010 and Staphylococcus, all within the top 20 genera. Two genera, W5053 and Lawsonella, were enriched in bulls of low fertility; this is the first time that these bacteria have been reported in bull semen samples. The majority of the bacteria were environmental organisms or were species originating from the mucous membranes of animals and humans. The results of this study indicate that differences in the seminal microbiota of healthy bulls occur and might be correlated with fertility. [ABSTRACT FROM AUTHOR]

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Copyright of Eurasian Journal of Veterinary Sciences is the property of Eurasian Journal of Veterinary Sciences and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background: Wide variation in fertility rates is observed when using frozen bull semen, even when the bulls have met quality standards for semen production. Therefore, a simple and reliable test to assess the freezing potential of bull semen based on the analysis of fresh semen or blood would be of great value. Attention is now turning to assessment of seminal plasma components such as proteins and elements. In the present study, the concentrations of macro- and microelements in fresh bull semen plasma and in serum and their correlation with quality characteristics of fresh semen and with semen quality after freezing and thawing were determined. Ejaculates were collected from 30 mature bulls, and semen volume, concentration, sperm motility, morphology, tail membrane integrity, plasma membrane permeability and DNA fragmentation were determined on the day of collection and after freezing and thawing. The concentrations of macroelements (Na, Mg, K and Ca) and microelements (Cu, Fe, Zn and Se) were determined in the seminal plasma and serum. The semen samples were classified into satisfactory and unsatisfactory groups according to the fresh semen quality. Results: Zinc and Se levels measured in serum were associated with almost all fresh and frozen-thawed semen quality characteristics, while Fe levels were associated only with acrosomal defects in fresh semen. Zinc and Fe levels in fresh seminal plasma were associated with various quality characteristics of fresh and frozen-thawed semen, while Se level in fresh seminal plasma was not associated with any of the semen quality characteristics. Conclusions: Microelements were shown to be useful as biomarkers involved in the analysis of bull sperm quality and could be used as an additional tool to predict bull semen quality after freezing and thawing. Our results confirm that the analysis of Zn and Se levels in serum and Zn, Cu and Fe levels in fresh seminal plasma can provide information to discriminate between bull semen samples with spermatozoa with high or low cryotolerance. [ABSTRACT FROM AUTHOR]

Contents: Breed and sire differences in sperm cryosurvival have been noted, with negative implications for sperm cryobanking and assisted reproduction programmes. This study hypothesized that these differences could be modified by using lower molecular weight cryoprotectants. Therefore, the effect of replacing glycerol (GLY) with ethylene glycol (EG) on differential cryosurvival of semen from two Sanga cattle breeds (Mashona vs. Tuli) was determined. Three to five ejaculates were collected from each of ten bulls (3‐8 years) by electro‐ejaculation, diluted in three Tris‐egg yolk extenders (Triladyl®, 7% GLY‐based and 7% EG‐based) and evaluated for sperm motility, viability and morphology at three time periods (fresh – 0 hr, pre‐freeze – 4 hr and post‐thaw). Tuli bulls produced larger (11.8 ± 0.31 ml vs. 8.5 ± 0.38 ml) and more concentrated ejaculates of lower fresh semen quality. Breeds differed across time for motility and morphology, but not viability. Mashona bull semen had significantly higher motility and normal morphology values at each sampling time. Bulls classified as poor freezers had lower concentration (0.70 ± 0.09 × 109 sperm/ml vs. 1.37 ± 0.10 × 109 sperm/ml), sperm motility index (SMI, 35.0 ± 3.4 % vs. 67.8 ± 2.1 %) and viability (69.7 ± 1.1 % vs. 75.7 ± 1.0 %) compared to good freezers. Maintenance of semen quality by GLY and EG did not differ between breeds, poor and good freezers, or age groups. The interaction breed by extender across time did not reach statistical significance for all variables. The study revealed that bull and breed variation in sperm quality and cryosurvival is not modified by replacing GLY with EG, suggesting that cryostress tolerance of sperm may be under control of mechanisms other than differential response to GLY cytotoxicity. [ABSTRACT FROM AUTHOR]

This study was designed to evaluate the effects of encapsulated reduced glutathione added to extender on the quality of bull sperm after freezing and thawing process. Extender contains lecithin nanoliposomes supplemented with different concenfrations of glutathione (0, 1, 2.5 and 5 Mm concenfration). The supplemented nanoliposomes with glutathione were prepared by thin-film preparation method. The particle size decreased to nanometer by sonication. 40 ejaculations were collected from 6 Holstein bulls and frozen for 6 weeks. Evaluated characteristics after freezing and thawing process were kinetic parameters (CASA), membrane activity (HOST), membrane integrity (eosinnigrosin) and morphology of sperm (Hancock's solution). Results showed that level 1 mM significantly increased the total and progressive motility (47.5±1.7, لح 32.72%). Although, there was no significant difference with level 2.5 mM glutathione (1.7 لح 45.02, لح 26.2%). Also, the concenfration of 2.5 mM showed the highest value of cell membrane integrity and cell membrane fimctionality but did not differ with the concenfration of ImM. However, supplementation of nanoliposomes with 5 mM glutathione reduced the quality of post-thaw sperm. Thus, level 1 M encapsulated glutathione can provide better protection compare to the other level of that for bull sperm. [ABSTRACT FROM AUTHOR]

Improving livestock reproductive performances is a high priority of worldwide breeders. The knowledge on cows accurate estrous detection associated with a good semen bull quality and artificial insemination appropriate timing, contribute in improving animal production yield. According to some studies, it is possible to achieve good quality semen from bulls by applying methods such the selection of bull presenting good conformation and health, followed by the collection and analysis of their semen, to be insured that this contains enough well fitted spermatozoids. For better success of artificial insemination, it is important to have a good mastery of the technique. The semen should also be protected against any agent of destruction and degeneration of spermatozoa, such as oxidizing processes. The present review gathers some technics of collecting and analyzing semen bull as well as some types of semen extender supplementation with antioxidant substances. [ABSTRACT FROM AUTHOR]

This study was designed to evaluate the effects of Treduced glutathione added to extender on the quality of bull sperm after freezing and thawing process. Extender contains lecithin nanoliposomes supplemented with different concentrations of glutathione (0, 1, 2.5 and 5 Mm concentration). The supplemented nanoliposomes with glutathione were prepared by thin-film preparation method. The particle size decreased to nanometer by sonication. 40 ejaculations were collected from 6 Holstein bulls and frozen for 6 weeks. Evaluated characteristics after freezing and thawing process were kinetic parameters (CASA), membrane activity (HOST), membrane integrity (eosinnigrosin) and morphology of sperm (Hancock's solution). Results showed that level 1 mM significantly increased the total and progressive motility (47.5±1.7, 32.7±2 %). Although, there was no significant difference with level 2.5 mM glutathione (45.0±1.7, 26.2±2 %). Also, the concentration of 2.5 mM showed the highest value of cell membrane integrity and cell membrane functionality but did not differ with the concentration of 1mM. However, supplementation of nanoliposomes with 5 mM glutathione reduced the quality of post-thaw sperm. Thus, level 1 M encapsulated glutathione can provide better protection compare to the other level of that for bull sperm. [ABSTRACT FROM AUTHOR]

Many microorganisms from various sources may be present in ejaculates of bulls. This study identified and isolated bacteria from bull sperm samples in a commercial stud and evaluated their resistance to antibiotics. The number of colony‐forming units was determined in semen samples collected at distinct steps during freezing and thawing. The minimum inhibitory concentration and the minimum bactericidal concentration were determined for four antibiotics commonly used in commercial studs. A total of 135 microorganisms from 25 genera were isolated. After a sensitivity test, all evaluated microorganisms (n = 55) were resistant to penicillin and most of them were resistant to tylosin and lincomycin (n = 54). Resistance to all tested antibiotics was observed in 22% of all isolates, whereas only 3.9% of the isolates were inhibited by the tested antibiotics at the concentrations recommended by the international legislation. As the isolated microorganisms presented high resistance to frequently used antibiotics, sensitivity tests should be periodically conducted in commercial bull semen studs to prevent the use of contaminated semen in artificial insemination. [ABSTRACT FROM AUTHOR]

The existing literature sans a collective picture of minerals and seminal plasma proteins in cow bulls having major sperm morphological defects. The present study involved six Jersey bulls imported from Denmark as calves. After sexual maturity, two bulls (Ab1 and Ab2; collectively as Ab) repeatedly exhibited abnormal spermiogram. The major sperm morphological abnormalities in Ab1 and Ab2, respectively, were Dag defect (32 and 27%), distal mid – piece reflex defect (42 and 17%) and coiled tail (11 and 12%). The mineral content of Ca and P was similar between the Ab and N (normal; N1, N2, N3 and N4 - collectively as N); values being higher in seminal plasma than blood plasma in both the groups. Seminal plasma of Ab had high K (indicates membrane damage), high Zn, especially in Ab2 (linked to Dag defect) and low Cu (decreases progressive motility). The seminal plasma proteins revealed absence of 15 kDa protein and weak expression of 25 kDa protein in Ab1. Both proteins are associated with sperm motility, viability and morphological abnormalities. In conclusion, the semen with abnormal sperm morphology also exhibited altered mineral content and seminal plasma proteins, which corroborates to their respective roles in seminology. [ABSTRACT FROM AUTHOR]

Freezing and thawing processes have the potential to adversely affect post-thaw quality of semen. In the current study, we endeavored to ascertain the most favorable application of L-cysteine for cryopreservation of Achai and Holstein Friesian bull semen under the subtropical conditions of Peshawar region of Pakistan. Semen of six adult bulls, three Achai (the indigenous breed) and three Holstein Friesian (exotic breed) amount to the experimental material at Government Cattle Breeding and Dairy Farm Harichand, Charsadda. Separate artificial vagina was utilized for semen collection from either breed, maintained at 42°C temperature. Gross and microscopic examination was carried out subsequent to the semen collection. Semen specimen with more than 70% motile characteristics were used for further processing. The semen was split into four aliquots in which tris-citric acid extender (TCA) was supplemented with various concentrations of L-cysteine 0 mM, 0.5 mM, 1.0 mM and 1.5 mM. Cooling and equilibration of the diluted semen was done at 4°C for 4 h subsequently filled in straws at the same temperature and were cooled in liquid nitrogen vapors for 10 min. followed by storage at -196°C for assessment. Progressive motility and viability of spermatozoa were significantly higher in 1.5 mM and 1.0 mM L-cysteine supplemented groups in comparison with 0.5 mM or 0 mM. The number of supravital hypo-osmotic swelling test positive spermatozoa at 1.5 mM and 1.0 mM was greater in comparison with lower concentration of 0.5 or 0 mM L-cysteine. Furthermore, significant improvement for acrosomal integrity was recorded in semen samples treated with 1.5 mM L-cysteine followed by 1mM, 0.5mM. It is concluded that 1.5 mM is the most favorable concentration of L-cysteine to be added to the TCA extender for improving the post-thaw quality of semen of Achai and Holstein Friesian bulls under the existing environmental conditions. [ABSTRACT FROM AUTHOR]

Our objective was to evaluate the effects of adding 50 mM trehalose (TRE), 1 mM cysteine (CYS), or 400 µg catalase (CAT), individually or in combinations, to a Tris-based semen extender and determine the effects on fresh, chilled-equilibrated, and frozenthawed bull sperm. Ejaculates from three Holstein bulls (1-3 years old) were pooled, divided into six equal portions, and put into a Tris-based extender supplemented with no additive (control), TRE, CYS, CAT, TRE + CYS, or TRE + CYS + CAT. In chilled-equilibrated semen, motility was highest in CAT but lowest in the control (P < 0.05). In thawed semen, motility was lower (P < 0.05) in CAT than in all other groups, which were not different. Based on a hypoosmotic swelling test (HOS test), TRE, CYS, and their combinations had higher (P < 0.05) percentages of sperm with an intact plasma membrane than the control or CAT groups. The TRE group had the least (P < 0.05) morphologically abnormal sperm. Plasma membrane and acrosomal integrity (PMAI) and high mitochondrial membrane potential (HMMP) values were highest in TRE and TRE + CYS and were lower (P < 0.05) in the control. Furthermore, HMMP was positively correlated with PMAI and HOS tests, but negatively correlated with major + minor abnormalities (r = 0.66, 0.242, and -0.349). In conclusion, the addition of TRE, with or without CYS, to a Tris-based semen extender improved several aspects of the frozen-thawed quality of bull semen. However, addition of CAT, by itself or in combination with other additives, did not protect bull sperm against cryopreservation-induced defects. [ABSTRACT FROM AUTHOR]

Copyright of Kafkas Universitesi Veteriner Fakultesi Dergisi is the property of University of Kafkas, Faculty of Veterinary Medicine and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Prediction of male fertility with in vitro semen quality parameters remains a challenge for the bull industry. Fluorescein staining and computer-assisted semen analysis (CASA) ensure kinetically proper and functionally objective results to improve spermatological parameter control. Therefore, we aimed to examine the CASA kinetic parameters and fluorescein staining of cryopreserved bull semen and its relationship with fertility. A total of 90 frozen straws from 5 Simmental bulls were evaluated. After semen was thawed, the motility, kinetic parameters, and insemination doses were evaluated by CASA; fluorescein stainings were evaluated using a fluorescein microscope. The 60th-day nonreturn rates (NRRs) were recorded for determination of pregnancy rates. There were significant differences in motility and kinetic parameters among bulls. Bull 3 had the lowest percentage of NRR and curvilinear velocity, straight line velocity, beat-cross frequency, and lateral head displacement parameters (P < 0.05). A positive correlation was detected between the pregnancy rate and some kinetic parameters. The correlation between the CASA parameters was determined as well. We concluded that various kinetic parameters obtained with CASA software system algorithms and fluorescein stainings can be linked to fertility. However, further research is needed to acquire a more precise link with fertility. [ABSTRACT FROM AUTHOR]

In this research, post-thaw effects of bull semen extenders including different amounts of boron (sodium pentaborate) on total motility (TMOT), progressive motility (PMOT), kinetic parameters of spermatozoa (VCL, VSL, VAP, LIN, STR, WOB, ALH, BCF), plasma membrane and acrosome integrity (PMAI) and DNA fragmentation index (DFI) were evaluated. Eight ejaculates were examined from four different bulls. Each ejaculate was split in to five aliquots and extended with standard Tris extender (Control) and 0.4 g boron with 0.5 g glucose (Group 1) 0.5; 0.7; 0.9 g boron without glucose (Group 2, Group 3 and Group 4 respectively) were used as experimental groups. After thawing TMOT, PMOT, kinetic traits of spermatozoa, PMAI and DFI were determined. According to motility and progressive motility (66.04%, 50.79%), Group 1 had higher values than the others, but differences between groups were not statistically significant (P>0.05). The highest values of VSL, LIN, STR and WOB were determined (28.15 µm/s, 42.98%, 71.69%, 59.71%) in Group 1 (P<0.05). While other kinetic traits of spermatozoa were higher numerically in Group 1 than the other groups, differences between groups were not statistically significant (P>0.05). The highest PMAI value was determined as 60.84% in Group 1 (P>0.05). The DFI rates were found similar in all control and experimental groups (P>0.05). In conclusion, it was observed that the boron (sodium pentaborate) has positive effects on post-thaw movement and structural traits. In addition, boron has no detrimental effect on DNA fragmentation index in semen freezing. Thus, boron could be used as an alternative semen extender compound. Özet: Bu çalışmada, farklı dozlarda bor (sodyum pentaborat) içeren boğa sperması sulandırıcılarının çözüm sonu toplam motilite (TMOT), progresif motilite (PMOT), spermatozoa hareket parametreleri (VCL, VSL, VAP, LIN, STR, WOB, ALH, BCF), plazma membranı ve akrozom bütünlüğü (PMAI) ve DNA fragmentasyon indeksi (DFI) parametreleri üzerine etkisi değerlendirildi. Dört baş boğadan toplam 8 ejakülat incelendi. Her ejakülat 5 eşit parçaya bölündü ve kontrol grubu olarak standart Tris sulandırıcısıyla sulandırıldı. Deney grubu sulandırıcıları olarak 0,5 g glikoza ek olarak 0,4 g bor eklenmiş Tris sulandırıcısı (Grup 1) ve glikoz yerine 0.5; 0.7; 0.9 g bor eklenmiş Tris sulandırıcıları (Grup 2, Grup 3, Grup 4) kullanıldı. Spermalar çözdürüldükten sonra TMOT, PMOT, spermatozoa hareket parametreleri, PMAI ve DFI değerleri belirlendi. Toplam motilite ve progresif motilite verilerine göre (%66.04, %50.79) Grup 1 sulandırıcısının daha yüksek değerlerde olmasına rağmen gruplar arası farklılıklar istatistiki açıdan önemsiz bulundu (P>0.05). En yüksek VSL, LIN, STR ve WOB değerlerinin (28.15 µm/s, %42.98, %71.69, %59.71) Grup 1'de olduğu görüldü (P<0.05). Diğer hareket özelliklerinin de Grup 1'de diğer gruplara göre yüksek olduğu belirlenirken, gruplar arası farklılıklar istatistiki açıdan önemsiz bulundu (P>0.05). En yüksek PMAI değerinin Grup 1'de olduğu, DFI değerlerinin ise tüm gruplarda benzer olduğu sonucuna ulaşıldı (P>0.05). Sonuç olarak borun çözdürme işleminden sonra spermatozoanın kinetik ve yapısal özellikleri üzerinde yararlı etkileri olduğu ortaya konmuştur. Ek olarak DFI açısından zararlı bir etkisi olmadığı gözlemlenmiştir. Bu veriler değerlendirildiğinde borun (sodyum pentaborat) alternatif bir sperma sulandırıcısı bileşeni olarak kullanılabileceği sonucuna ulaşılmıştır. [ABSTRACT FROM AUTHOR]

Various foot-and-mouth disease (FMD) virus strains circulate in the Middle East, causing frequent episodes of FMD outbreaks among Israeli livestock. Since the virus is highly resistant in semen, artificial insemination with contaminated bull semen may lead to the infection of the receiver cow. As a non-FMD-free country with vaccination, Israel is currently engaged in trading bull semen only with countries of the same status. The purpose of this study was to assess the risk of release of FMD virus through export of bull semen in order to estimate the risk for FMD-free countries considering purchasing Israeli bull semen. A stochastic risk assessment model was used to estimate this risk, defined as the annual likelihood of exporting at least one ejaculate of bull semen contaminated with viable FMD virus. A total of 45 scenarios were assessed to account for uncertainty and variability around specific parameter estimates and to evaluate the effect of various mitigation measures, such as performing a preexport test on semen ejaculates. Under the most plausible scenario, the annual likelihood of exporting bull semen contaminated with FMD virus had a median of 1.3 * 10−7 for an export of 100 ejaculates per year. This corresponds to one infected ejaculate exported every 7 million years. Under the worst-case scenario, the median of the risk rose to 7.9 * 10−5, which is equivalent to the export of one infected ejaculate every 12,000 years. Sensitivity analysis indicated that the most influential parameter is the probability of viral excretion in infected bulls. [ABSTRACT FROM AUTHOR]

This the study was conducted to find out the effect of the addition of alpha linolenic acid (ALA) and docosahexaenoic acid (DHA) on frozen-thawed quality of bull semen. Twenty-four ejaculates (8 from each bull) were collected from three bulls. Semen ejaculates having motility .70% and normal morphology .80% were extended into Tris extender consist of equal concentration of each ALA and DHA (0, 3, 5, 10,15ng/ml). Extended semen was cooled at 5oC for 2 hours at 5oC and then packaged in 0.25ml straws and stored into liquid nitrogen for 24 hours. Successively, thawing and evaluation was performed for sperm motility, viability, membrane integrity, morphology, malondialdehyde (MDA), fatty acid composition and super oxide dismutase (SOD). Sperm characteristics such as sperm motility, viability, morphology, membrane integrity, and acrosome integrity were decreased in treated groups. DHA concentration was increased in all supplemented groups. Whereas ALA decreased in all groups except at 5ng/ml DHA was increased linearly by increasing supplementation of fatty acids. In conclusion, combination of ALA and DHA did not improve frozen-thawed bull sperm parameters when compared to control. [ABSTRACT FROM AUTHOR]

Copyright of Kafkas Universitesi Veteriner Fakultesi Dergisi is the property of University of Kafkas, Faculty of Veterinary Medicine and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Animal & Poultry Production is the property of Egyptian National Agricultural Library (ENAL) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Commercially available OptiXcell® extender was compared with conventional extenders for freezability and in vivo fertility of bull semen. Semen was collected from three Friesian bulls for five weeks (replicate) and qualifying ejaculates (motility >60%, concentration >0.5 billion/mL, volume >1mL) were diluted (37°C; 50 × 106 spermatozoa/ml) with OptiXcell®, tris-citric egg yolk and egg yolk-citrate extenders. Diluted semen was cooled to 4°C in 2 hours, equilibrated for 4 hours and filled in 0.5 ml straws. The straws were kept over liquid nitrogen vapours for 10 minutes and plunged into liquid nitrogen. Percentages of post thaw sperm motility and plasma membrane integrity were recorded higher (P<0.05) in OptiXcell® compared to tris-citric egg yolk followed by egg yolk-citrate extender. Sperm viability (%) were recorded higher (P<0.05) in OptiXcell® compared to tris-citric egg yolk and egg yolk citrate extender. Percentages of normal apical ridge and DNA integrity were higher (P<0.05) in OptiXcell® and tris-citric egg yolk extender compared to egg yolk-citrate extender. Higher (P<0.05) fertility rate was recorded with semen frozen in OptiXcell® compared to triscitric egg yolk and egg yolk-citrate extender. In conclusion, OptiXcell® is superior to conventional extenders for spermatozoal quality of frozen-thawed bull semen and produced higher fertility rates under field conditions. [ABSTRACT FROM AUTHOR]

Proteins are the most important organic substances which are part of the composition of living cells. Since the specific capabilities of spermatozoa depend on the protein, occurs the necessity of studying the characteristics of changes of the spectrum of the protein fractions of sperm depending on the ambient temperature. In this experiment was studied the effect of temperature of 45°C on the spectrum of protein fractions of the bull semen. From the data of the conducted research it is observed that when sperm of bull was exposed to a temperature of 45°C for 1 minute, has been a trend of increasing the quantity of albumin up to 4.2% and at 10 minutes - up to 3.3% of the total volume. When exposed to a temperature of 45°C for 1 minute the total amount of globulins, typically does not significantly change. The content of a-globulins tends to decrease, while the ß- and ?-globulins do not support the changes. After 10 minutes of stress the amount of albumins is also increasing in comparison with the control sample and the quantity of total globulins decreased, especially a - and ?-globulins, while the content of ß-globulin is not changed. Thus, the hyperthermic stress of bull semen for 1 minute is reflected only on the quantity of albumin, which has a tendency to increase. The influence of hyperthermic stress for 10 min is manifested by the tendency of growth of the percentage content of albumin and insignificant decrease in the concentration of total globulins, including a-globulin. [ABSTRACT FROM AUTHOR]

In this study, paraoxonase 1 (PON1; EC 3.1.8.1) was purified from bull semen, and some characteristics of the enzyme were investigated. In vitro inhibition effect of some heavy metals, including Cu, Mn, Cd, Zn, Ni, and Pb, on the activity of the purified enzyme was also investigated. The purification of bull semen PON1 procedure was composed of two steps: ammonium sulfate precipitation and Sepharose-4B- l-tyrosine-1-naphthylamine hydrophobic interaction chromatography. The enzyme, having a specific activity of 288 EU/mg proteins, was purified 22.67-fold with a yield of 89 %. Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis of the purified enzyme showed the presence of a single band with an apparent MW of 66 kDa. The V and K values for the paraoxon substrate were determined as 100 EU and 8.0 × 10 M, respectively. The inhibitory effects of different heavy metals on PON1 activity were determined by using the paraoxon as a substrate. The results showed that all the metals, except for Cd, inhibited the PON1 enzyme activity in a concentration-dependent fashion. IC values of Cu, Mn, Zn, Ni, and Pb were found as 2.59 × 10, 1.17 × 10, 42.74 × 10, 99.10 × 10, 48.80 × 10 mM, respectively. Conversely, Cd increased the bull semen PON1 enzyme activity. The present study has demonstrated that Cu, Mn, Zn, Ni, and Pb are serious toxic metals, which are able to increase the risk of oxidative stress development and a subsequent decrease of semen quality. [ABSTRACT FROM AUTHOR]

Animal origin egg yolk extender has been associated with microbial contamination and interference with microscopic examination. Therefore, replacement of such extender with plant origin compound is inevitable without compromising the desired qualities of an ideal extender. In the current experiment, we compared the effect of soy-based extender (25% and 50%) of five adult Holstein-Friesian bulls semen preserved at 4°C for 24 h with conventional egg yolk (control) and synthetic extender. Semen quality was assessed by measuring liveability, individual motility and membrane integrity after 24 h of extended semen stored at 4°C. The results indicated that liveability, motility of sperm and membrane integrity decreased significantly (P<0.05) in soy 50% extender. The results of sperm liveability, individual motility, and membrane integrity in 25% soy extender were comparable with the control. Therefore, we concluded that 25% soy milk could be used as a substitute of conventional egg yolk-based extender for bull semen stored at 4°C. [ABSTRACT FROM AUTHOR]

Infection with Bluetongue virus (BTV) is a significant impediment to the global movement of bovine semen. Repeat testing of blood from donor animals is specified in the World Organization for Animal Health (OIE) Manual for the export of semen from regions where BTV may be present. Screening of blood or semen samples has usually been carried out by virus isolation (VI) either by inoculation of chicken embryos followed by passage onto insect and mammalian cell cultures or in vivo inoculation of sheep followed by serology to detect seroconversion. Direct testing of semen for BTV would enable earlier release of semen samples and avoid repeat testing of the donor, as well as provide an option for releasing batches of semen that were collected without certification of the donor. Quantitative (real-time) reverse transcription polymerase chain reaction (qRT-PCR) assays overcome most of the limitations of other methods and have the potential to provide higher sensitivity. The present study compared 5 qRT-PCR assays, including 2 commercially available kits, for the detection of BTV in semen serially collected from 8 bulls over a period of 90 days after experimental infection. The results of the study show that at least one of the qRT-PCR assays is extremely reproducible and has both very high sensitivity and specificity to reliably detect all available serotypes. The preferred qRT-PCR gave consistently superior results to VI, sheep inoculation, and conventional RT-PCR. Therefore, the assay can be recommended for the screening of bovine semen for freedom from BTV. [ABSTRACT FROM AUTHOR]

The effect of reduced glutathione (5 mM) and cysteine hydrochloride (5 mM) on sperm velocity and motility parameters of good (3) and poor (3) freezable Jersey crossbred bull semen was studied. The post-thawed sperm velocity and motility parameters were assessed by computer assisted sperm analyzer (CASA) at different hours of incubation. The forward progressive and total motility showed significant (p<0.05) difference in glutathione treated group as compared to cysteine treated and control group in both freezer. In both freezers, the forward progressive and total motility was not decreased significantly up to 2 h of incubation but thereafter there was a highly significant decrease. A gradual non-significant decline in all the velocity parameters in good freezable semen samples were observed for the 4 periods of incubation and in poor freezable semen, the reduction in the velocity parameters was not significant up to 2 h of incubation, but thereafter, the decrease was significant. The velocity parameters of sperm revealed that the glutathione treated group was non-significantly higher than cysteine treated and control group in both good and poor freezable bulls except the addition of glutathione significantly improved VCL (curvilinear velocity) and ALH (amplitude of lateral head displacement) of the post-thawed spermatozoa at 4 h of incubation. Further, the addition of cysteine and glutathione has non-significantly improved the other velocity parameters in both the freezer. Thus, the reduced glutathione has improved the post-thawed sperm functional parameters such as motility and velocity in good and maintained in poor freezable semen. [ABSTRACT FROM AUTHOR]

The study was conducted to evaluate the effect of addition of L-cysteine to tris-citric acid (TCA) extender on the post-thaw quality of Sahiwal bull semen. For this purpose, two consecutive ejaculates were collected from three Sahiwal bulls using artificial vagina at weekly intervals for a period of three weeks (three replicates). Qualifying semen ejaculates were diluted (50 x 106 motile spermatozoa ml-1) in TCA extender having L-cysteine either 0.0 (control) or 0.5, 1.0 or 2.0 mM. Diluted semen was cooled to 4°C for 2 h, equilibrated for 4 h at 4°C, filled in straws at 4°C, kept in liquid nitrogen vapours for 10 min and then stored in the liquid nitrogen. Thawing was performed after 24 h of storage, at 37°C for 30 s. and the sperm motility, viability, plasma membrane and acrosomal integrity were assessed. Higher (P<0.05) sperm motility, viability, plasma membrane and acrosomal integrity were observed using extenders containing 1.0 or 2.0 mM compared to those containing 0.5 or 0.0 mM L-cysteine. It is concluded that addition of L-cysteine (to reach 1.0-2.0 mM) in TCA extender improves the post-thaw quality of Sahiwal bull semen. [ABSTRACT FROM AUTHOR]

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The objective of the present study was to confirm the presence of prematurely capacitated spermatozoa in frozen–thawed bull semen and to investigate the relationship of premature capacitation to the fertility of the respective semen. Twenty batches of frozen semen from young AI bulls of the Swedish Red and White breed with known fertility (expressed as 56-day non-return rates; 56 d-NRR) were tested using a Chlortetracycline (CTC) assay to assess capacitation status in frozen–thawed spermatozoa. The status of capacitation, as evidenced in this experiment, was further tested based on the hypothesis that capacitated spermatozoa present in frozen–thawed semen should undergo the acrosome reaction (AR) on co-incubation with homologous zona pellucida (ZP) glycoproteins. The percentages (mean ± SEM) of uncapacitated, capacitated and acrosome-reacted spermatozoa in the frozen–thawed semen (n equals; 20) were 49.3 ± 11.9, 36.3 ± 8.3 and 14.2 ± 11.9, respectively. On co-incubation with ZP, there was a significant increase (p equals; 0.001) in the proportion of spermatozoa undergoing the AR compared to the control with a concurrent decrease in the proportion of capacitated spermatozoa, suggesting that a proportion of capacitated spermatozoa were undergoing the AR. The proportion of viable, uncapacitated spermatozoa present in the frozen–thawed semen was correlated to the 56 d-NRR (n equals; 20, r equals; 0.5, p equals; 0.03). In conclusion, a proportion of spermatozoa in frozen–thawed semen was capacitated and the proportion of viable, uncapacitated spermatozoa present in semen was positively correlated to fertility. [ABSTRACT FROM AUTHOR]

Simple Summary: Oxidative stress in mammalian sperm can be induced during cryopreservation; therefore, it is important to inhibit oxidative stress to maintain sperm motility during cryopreservation. The present study was performed to investigate the effects of supplementing apigenin (AP), astragalus polysaccharide (APS), or their combination on an extender for bull semen freezing. Sperm viability, motility, and motion parameters; acrosome integrity and membrane integrity; endogenous antioxidant indices; and reactive oxygen species (ROS) and malondialdehyde (MDA) levels were evaluated after semen thawing. Our results indicated that 0.2 mmol/L AP or 0.5 mg/mL APS improved the cryopreservation of bull sperm; moreover, their combination in the extender could further improve the protective effects on bull sperm post-thaw. The purpose of this study was to determine the effects of apigenin and astragalus polysaccharides on the cryopreservation of bovine semen. Apigenin, astragalus polysaccharides, or their combination were added to a frozen diluent of bovine semen. Afterwards, Computer Assisted Semen Analysis (CASA), membrane functionality, acrosome integrity, mitochondrial integrity, CAT, SOD, GSH-Px, MDA, and ROS detection were conducted. The results showed that adding 0.2 mmol/L AP or 0.5 mg/mL APS could improve the quality of frozen sperm. Compared to 0.2 mmol/L AP alone, the combination of 0.2 mmol/L AP and 0.3 mg/mL APS significantly increased the total motility (TM), average path distance (DAP), straight line distance (DSL), average path velocity (VAP), curvilinear velocity (VCL), wobble (WOB), and sperm CAT and SOD levels (p < 0.05), while reducing the ROS and MDA levels (p < 0.05). These results indicated that the addition of 0.2 mmol/L AP or 0.5 mg/mL APS alone has a protective effect on the freezing of bovine semen. Compared to the addition of 0.2 mmol/L AP, a combination of 0.2 mmol/L AP and 0.3 mg/mL APS could further improve the quality of frozen semen. [ABSTRACT FROM AUTHOR]