Search

Abstract The reproductive performance during the transition from prepubertal to pubertal stage was evaluated in Osmanabadi breed bucks supplemented with organic Zinc (Zn) and Copper (Cu). A total number of 40 bucks aged 20 weeks were randomly assigned to 10 groups (each n = 4). The control group was maintained with basal diet, without any additional mineral supplementation. The treatment groups were supplemented with graded doses of organic Zn (Zn 20 mg, Zn 40 mg and Zn 60 mg), Cu (Cu 12.5 mg, Cu 25 mg and Cu 37.5 mg) and a combination of Zn + Cu (Zn 20 mg + Cu 12.5 mg, Zn 40 mg + Cu 25 mg and Zn 60 mg + Cu 37.5 mg), respectively for a period of 26 weeks (up to the age of 46 weeks). Sexual behaviour and scrotal biometry were recorded periodically. Blood and semen samples were collected and processed for LH estimation in blood plasma, and testosterone, T3 and T4 hormones in the seminal plasma. The mounts with ejaculation were observed earlier (P < 0.05) in the treatment bucks (from 38th week of age) than the control group (43rd week onwards). A positive correlation was observed between blood plasma LH and testosterone with total mounts (r = 0.31, P < 0.05; r = 0.51, P < 0.01) and mounts without ejaculation (r = 0.40, P < 0.01; r = 0.52, P < 0.01). A negative correlation between T4 with sperm number per ejaculation (r = −0.31, P < 0.05) and sperm concentration (r = −0.35, P < 0.05) had been noticed. Different doses of minerals showed positive interaction (P < 0.05) with sperm functional and behavioural characteristics. The spermatozoal gene expression of ODF2 and ZCCHC6 were significantly influenced by the mineral supplementation in all doses. The ZCCHC6 gene expression was positively correlated with testosterone (r = 0.50, P < 0.001) and sperm number per ejaculation (r = 0.42, P < 0.001), and ODF2 gene with T3 hormone (r = 0.34, P < 0.05). The present study indicates that the diet supplemented with organic trace minerals cause intense sexual behaviour, enhancement in sperm number per ejaculate, total motility, spermatozoal genes expression and altered LH, testosterone and T4 hormones. Graphical abstract Image 1 Highlights • First study regarding the role of Zn and Cu on male sexual behavior in goats. • Mineral supplementation influences LH, testosterone, T3 and T4 hormone levels. • Zn and Cu modulates the sperm quantitative and qualitative characteristics. • Mineral supplementation improved the male fertility genes (ODF2 and ZCCHC6) expression in spermatozoa. [ABSTRACT FROM AUTHOR]

Abstract The antioxidant properties and the protective role of organic zinc (Zn) and copper (Cu) in white blood cells (WBCs) and spermatozoa were analyzed through quantification of superoxide dismutase 1 (SOD1), catalase (CAT), glutathione peroxidase 4 (GPx4) and nuclear factor erythroid 2-like 2 (NFE2L2) and correlations were determined with sperm functional characteristics in Osmanabadi bucks. Bucks (aged 5 months; n = 40) were divided into ten groups, and the dietary treatments comprised of a control and nine treatment groups as follows: organic Zn as Zn 20, Zn 40 and Zn 60, organic Cu as Cu 12.5, Cu 25, Cu 37.5 and combined organic Zn and Cu as Zn 20+Cu 12.5, Zn 40+Cu 25, Zn 60+Cu 37.5, respectively per kg dry matter for a period of 8 months. The blood (120 and 240 days) and semen (240 days: 40 × 4 = 160) samples were collected from 40 bucks. In WBCs: the relative abundance of mRNA for SOD1, CAT, GPx4, NFE2L2 was greater (P < 0.05) in (120 and 240 days) in majority of the mineral supplemented animals. In spermatozoa: the relative abundance of SOD1, NFE2L2, GPx4 and CAT mRNA was greater (P < 0.05) in selected treatment groups. The abundance of SOD1 mRNA in WBCs was positively correlated (P < 0.05) with sperm mass motility (r = 0.692, P = 0.027). The abundance of GPx4 mRNA was negatively correlated (P < 0.05) with type A sperm (straightness; STR) > 85% and amplitude of lateral head displacement (ALH) > 2.5 μm/ s) (r = −0.711, P = 0.021) and (P < 0.05) positively correlated with sperm viability (r = 0.669, P = 0.035). Organic Zn and Cu supplementation was associated with an increase in the expression of antioxidant defense enzyme genes in bucks. [ABSTRACT FROM AUTHOR]

Attainment of puberty in animals is dependent on their age, body weight, nutritional status, genetic and environmental conditions. Nutritionally, organic minerals are suggested to improve semen production, sperm motility and male fertility. In this context, role of organic zinc (Zn) and copper (Cu) in advancing male puberty and semen characters in Osmanabadi goats were studied. Forty one (n = 41) bucks (Aged 5 months) were divided into ten groups and the dietary treatments comprised of a control group (basal diet; without additional trace mineral supplementation) and nine treatment groups that received, in addition to the basal diet, various doses of trace minerals (mg) on per kg dry matter basis, organic Zn as low Zn20, medium Zn40 and high Zn60, organic Cu as low Cu12.5, medium Cu25, high Cu37.5 and combination of organic Zn + Cu as low Zn20 + Cu12.5, medium Zn40 + Cu25, high Zn60 + Cu37.5, respectively fed for a period of 8 months. Bucks fed organic trace minerals reached puberty 28–35 days earlier than control group. In addition, improvement ( P < .01) in testosterone hormone (ng/ml) levels (control: 1.63 ± 0.07 VS Zn60: 2.54 ± 0.02; Cu12.5: 6.17 ± 0.05; Cu25: 3.01 ± 0.04; Cu37.5: 2.39 ± 0.06; Zn20 + Cu12.5: 1.94 ± 0.02; Zn60 + Cu37.5: 2.44 ± 0.16 at 240 days), semen production capacity (sperm concentration, volume, mass motility) and semen quality (higher progressive motility, velocity, sperm membrane integrity and acrosome integrity) were observed in supplemented groups ( P  < .05) than the control bucks. The present study demonstrated that, additional feeding of organic Zn and Cu to growing male goats advanced onset of puberty and improved quantitative and qualitative semen characteristics. The results also implied that the organic Cu had a significant effect on overall performances of bucks as compared to Zn alone or Zn and Cu in combination. [ABSTRACT FROM AUTHOR]

The possibility of including amino acids for cryopreservation of ram semen to improve the quality of frozen semen was explored in this study in sheep model. 24 samples were collected in triplicate from 8 rams of 2–3 year old Bannur cross bred rams maintained at the Institute Experimental Livestock Unit. Semen was diluted in tris-egg yolk glycerol diluent and made into 7 aliquots as follows: aliquot 1 served as control, “ l -alanine” was added at 100 and 135 mM in the aliquots 2 and 3, “ l -glutamine” was added at 20 and 25 mM in the aliquots 4 and 5 and “ l -proline” was added at 25 and 50 mM in the aliquots 6 and 7, respectively. Diluted semen was filled in 0.25 ml French straws and frozen in LN 2 . Inclusion of “ l -proline” and “ l -glutamine” in the diluent increased the percent live sperm ( P < 0.001), total motility ( P < 0.05) and maintained higher functional membrane and acrosomal integrity ( P < 0.001) by decreasing lipid peroxidation ( P < 0.001) compared to the control group. In contrast, “ l -alanine” decreased the percentage of total motility, fast progressive spermatozoa and increased ( P < 0.01) the percentage of immotile spermatozoa. It can be concluded that 20 mM “ l -glutamine” and 25 mM “ l -proline” can be used as semen additive to freeze ram semen as they prevented cryoinjuries to sperm and improved the pre-freeze and post-thaw semen characteristics. [ABSTRACT FROM AUTHOR]

Abstract: The objective was to determine the association of mRNA expression of cystine rich secretary protein 2 (CRISP2), chaperonin containing T-complex protein 1, subunit 8 (CCT8), and phosphatidylethanolamine-binding protein 1 (PEBP1), in sperm of Holstein bulls with Sire Conception Rate (SCR) scores between −4 and +4. These proteins were involved in sperm capacitation and sperm-egg fusion. Samples of sperm obtained on a single day from Holstein bulls (N = 34) in a commercial AI centre were used to evaluate relative mRNA expression of CRISP2, CCT8, and PEBP1. The mRNA abundance of CRISP2 was positively correlated (r = 0.88; P < 0.002), CCT8 was negatively correlated (r = −0.87; P < 0.002), and PEBP1 was positively correlated (r = 0.83; P < 0.006) with SCR-scores. The means of CRISP2 mRNA abundance was greater among positive SCR-score bulls (2.5 to 8 fold), the means of CCT8 mRNA abundance was greater among the negative SCR-score bulls (9.5 to 3.5 fold), and the means of PEBP1 mRNA abundance was greater for the positive SCR-score bulls (5.4 to 7.7 fold). In multivariate regression models predicting SCR-scores, mRNA abundance of CCT8 was significantly associated with SCR-score in all models. In the presence of CRISP2 mRNA abundance in the model, the SCR score''s predictability of PEBP1 was insignificant. However, in the absence of CRISP2 mRNA abundance in the model, the SCR-score''s predictability of PEBP1 was significant. In multivariate regression models, CRISP2 and CCT8 mRNA expression in sperm accounted for 95% of the variance in Holstein bull''s SCR-scores. In conclusion, Holstein bulls with greater CRISP2 and lower CCT8 mRNA expression in sperm had higher probabilities of siring calves. [Copyright &y& Elsevier]

Abstract: Seventy semen ejaculates were obtained from 14 Murrah buffalo bulls and were subjected to plasma separation immediately after collection by centrifugation at 2000rpm for 20min and stored in liquid nitrogen until analysis. In the seminal plasma the total protein concentration were estimated and the heparin and gelatin binding (HB and GB) proteins were isolated using heparin and gelatin affinity column chromatography. The molecular weight of individual isolated HB and GB protein was determined by SDS–PAGE analysis. Buffalo bull spermatozoa was collected from cauda epididymis under aseptic conditions and was used for the in vitro fertility tests (i.e. bovine cervical mucus penetration test (BCMPT) and hypo-osmotic swelling test (HOST)). The heparin and gelatin binding buffalo seminal plasma proteins were used in six concentrations i.e. 10, 20, 30, 40, 50 and 60μg/ml to test their effect on in vitro fertility assessment of cauda epididymal spermatozoa. The overall mean values of total protein, HB and GB proteins were recorded as 29±2.7, 2.61 and 0.2mg/ml, respectively. Eighteen total protein bands were observed in the range of 12–127kDa. Eight major HB proteins were isolated in the range of 13–71kDa. Seven major GB proteins were isolated in the range of 13–61kDa in the buffalo seminal plasma. The mean penetration distance (mm) travelled by the buffalo cauda spermatozoa was maximum in HB proteins (26.9±0.6) followed by GB proteins (25.4±0.6) and control (21.2±1.4). The difference in BCMPT values between protein treated and control group was significant (P <0.05). Almost similar trend in the effect of protein on values of HOST percentage in both HB and GB proteins treated semen samples were recorded (66.4±0.65 and 66.1±0.6, respectively). The difference in HOST values between proteins treated and control group (50.4±2.0) was significant (P <0.05). The present results indicate that among the isolated proteins, 4 proteins were commonly seen in both the heparin and gelatin–sepharose affinity column chromatography, and the addition of buffalo seminal plasma proteins improved the in vitro sperm functions (40μg/ml gave best results) of buffalo cauda spermatozoa. [Copyright &y& Elsevier]

Abstract: Two ultrasound cases are presented. In case one, a 19-year-old male complained of a testicular lump on the left side. Ultrasound revealed three separate testes and epididimi, consistent with the rare triorchidism anomaly, one of the presentations of polyorchidism. In case two, a 24-year-old male complained of left peri-testicular swelling. Ultrasound revealed an unusual cleft centrally within the left testis extending for over 50% of the testis. The testicular cleft anomaly was thought to be a variation of the rare triorchidism anomaly previously unreported in the literature. Both cases appear to be within the same spectrum of rare scrotal anomaly. [Copyright &y& Elsevier]

The objective of this study was to assess the influence of spermatozoa surface antigenic proteins on the functional competence of bovine neat and frozen-thawed semen. The breeding bulls (n = 38) were screened for seminal antigenic levels in neat semen based on the agglutination titrations with anti-sperm antibody (ASA). Bulls having high (n = 8) and low (n = 7) antigenic levels were selected and spermatozoa functional parameters were analyzed in neat and frozen-thawed semen samples. In neat semen, kinematics such as straightness (73.6 ± 1.0 and 66.9 ± 1.5%), linearity (48.6 ± 1.2 and 40.1 ± 3.9%), curvilinear velocity (103.3 ± 2.6 and 93.4 ± 3.8 μm/s), straight-line velocity (65.7 ± 2.6 and 53.7 ± 2.2 μm/s) and average path velocity (53.8 ± 2.5 and 39.8 ± 2.3 μm/s) were significantly high (p < 0.05) in samples with lower antigenicity. The percentage of spermatozoa that can penetrate mucus (49.9 ± 2.3 and 37.1 ± 3.2) was significantly higher in semen samples with low ASAs. The total motile (84.0 ± 2.5 and 86.0 ± 1.5) and progressive motile (68.4 ± 3.7 and 69.2 ± 1.6) spermatozoa were higher in neat semen samples with higher antigenicity. A significantly (p < 0.05) higher mitochondrial membrane potential was observed in neat (82.5 ± 2.8 and 69.0 ± 2.0%) and post-thaw (28 ± 5. 6 and 16 ± 3.7%) samples of the lower antigenic group. The percentage of acrosome-reacted spermatozoa was significantly (p < 0.05) higher in neat (58.7 ± 2.9 and 52.6 ± 1.8), but reduced significantly (p < 0.05) in post-thaw (32.0 ± 2.0 and 48.0 ± 2.6) semen of higher antigenic groups. The study reveals that higher seminal antigenicity reduces mitochondrial membrane potential and acrosome reaction ability in post-thaw spermatozoa. • Higher levels of seminal antigen affect spermatozoa kinematics and mitochondrial membrane potential in neat semen. • Levels of seminal antigens did not influence spermatozoa kinematics in post-thaw semen samples. • Seminal antigenicity did not influence spermatozoa structural and functional membrane integrities during cryopreservation. • Higher levels of seminal antigens prevent acrosome reaction in post-thaw spermatozoa. • Antigenic levels in semen may affect the fertilization competence of frozen semen samples. [ABSTRACT FROM AUTHOR]

Abstract Sperm are highly specialized compartmentalized cells, with unique compositional, morphological and functional properties, including a plasma membrane that undergoes dynamic protein remodeling and surface modifications. Seminal plasma is a highly complex biological fluid containing proteins, amino acids, enzymes, fructose and other carbohydrates, lipids, major minerals and trace elements. Seminal plasma proteins are involved in regulation of osmotic pressure and pH of seminal plasma, transport of ions, lipid and hormones. The objective was to compare sperm and seminal plasma proteomes of bulls with differing fertility and to relate differences to biological processes. Semen was collected from bulls with high or low fertility (4 bulls in each category). Sperm and seminal plasma proteins were isolated, purified, subjected to 2-D gel electrophoresis, protein identification and ontology. In sperm and seminal plasma, binder of sperm proteins (BSP)-1, −3 and −5, and spermadhesin-1, ALB, TIMP, AKI and PEBP1 were higher for high-versus low-fertility bulls (P < 0.05), whereas proteins CLU, CCT5 and 8, ELSPbP1, and PSMA6 were more abundant in sperm and seminal plasma of low- versus high-fertility bulls (P < 0.05). Further, HSP90, ZFP34, IFNRF4, BCL62, NADHD, TUBB3 and Histone H1 were in greater abundance in sperm of high- compared with low-fertility bulls. The two key biological processes of proteins differentially expressed in high- and low-fertility bulls were metabolic processes and biological regulation. The most prominent molecular functions for proteins that differed are binding, catalytic and receptor activities. The main cellular components for proteins that differed are cellular, extracellular, and plasma membrane. Since protein content differed in high- versus low-fertility bulls, we inferred that the efficiency of associated sperm functions that are necessary for fertility may also differ between high- and low-fertility semen. In conclusion, differences between high- and low-fertility bulls regarding abundance of sperm and seminal plasma proteins likely contributed to differences in fertility. Highlights • BSP-1, -3, -5, and spermadhesin-1 were in greater abundance in sperm and seminal plasma of high-fertility Holstein bulls. • Isoforms of CLU, ALB, TIMP, AK1 and PEBP1 were in greater abundance in sperm and seminal plasma of high-fertility bulls. • HSP90, ZFP34, IFNRF4, BCL62, NADHD, TUBB3 and Histone H1 were in greater abundance in sperm of high-fertility bulls. • Clusterin, CCT5 and 8, ELSPbP1, and PSMA6 were in greater abundance in sperm and seminal plasma of low-fertility bulls. • PANTHER identified differed biological processes, molecular functions and cellular components for differed proteins. [ABSTRACT FROM AUTHOR]

Mammalian sperm are exposed to a natural hypoosmotic environment during male-to-female reproductive tract transition; although this activates sperm motility in vivo , excessive swelling can harm sperm structure and function. Aquaporins (AQPs) is a family of membrane-channel proteins implicated in sperm osmoregulation. The objective was to determine associations among relative sperm volume shift, hypoosmotic swelling test (HOST), sperm aquaporin (AQP) 7 mRNA abundances, and sire conception rate (SCR; fertility estimate) in Holstein bulls at a commercial artificial insemination center. Three or four sires for each full point SCR score from −4 to +4 were included. Each SCR estimate for study bulls (N = 30) was based on > 500 services (mean ± SEM) of 725 ± 13 services/sire). Sperm from a single collection day (two ejaculates) from these commercial Holstein bulls were used. Relative mRNA expression of AQP7 in sperm was determined by polymerase chain reaction. Mean relative sperm volume shift and percentage of sperm reacted in a HOST (% HOST) were determined (400 sperm per bull) after incubating in isoosmotic (300 mOsm/kg) and hypoosmotic (100 mOsm/kg) solutions for 30 min. There was no correlation between %HOST and SCR (r = 0.28 P > 0.1). However, there was a positive correlation between relative sperm volume shift and SCR (r = 0.65, P < 0.05). Furthermore, AQP7 mRNA abundance was positively correlated to both relative volume shift (r = 0.73; P < 0.05) and to SCR (r = 0.67; P < 0.05). The mRNA expressions of AQP7 and relative sperm volume shift differed (P < 0.05) among low- (<2 SCR), average- (-2 to +2) and high- (>2) fertility sire groups. In conclusion, bulls with higher SCR had significantly greater AQP7 mRNA abundance in frozen-thawed sperm. This plausibly contributed to greater regulation of sperm volume shift, which apparently conferred protection from detrimental swelling and impaired functions. [ABSTRACT FROM AUTHOR]

Abstract: The study was conducted to assess the effect of heparin binding seminal plasma proteins (HBP) on freezability and in vitro fertilizing ability of buffalo cauda epididymal spermatozoa. Spermatozoal motility, viability and acrosomal integrity at prefreeze and post-thaw stages were studied. The in vitro fertilizing ability of spermatozoa was assessed by the application of two tests, i.e. bovine cervical mucus penetration test (BCMPT) and hypo-osmotic sperm swelling test (HOST). HBP isolated from buffalo seminal plasma and maintained in the laboratory were used for the study. Twelve pairs of epididymis from adult buffaloes slaughtered at the local abattoir were used for the study. The results indicated that HBP addition improved the progressive motility, BCMPT and HOST response at prefreeze level. HBP at a concentration of 40μg/ml showed better results than HBP at a concentration of 80μg/ml. However, subjecting the HBP treated spermatozoa to cryopreservation resulted in significant reduction of motility, viability, acrosomal integrity and response to BCMPT and HOST in the HBP treated groups when compared to those in control group. The deleterious effect of HBP was found to be concentration dependent with the higher concentration causing higher post-thaw damage. [Copyright &y& Elsevier]

To test our hypothesis that antisperm antibodies (ASA) might alter sperm phenotypic attributes thus leading to sub-fertility/infertility in bulls, ASA were generated in crossbred male calves by immunizing with sperm two times. Cryopreserved spermatozoa from crossbred bulls (n = 24) with different field fertility ratings were incubated with ASA and different patterns of ASA immunolocalization were studied. In addition, sperm membrane integrity, acrosomal integrity and cryo-capacitation status were also assessed. Immunolocalization of sperm antigens using antisperm antibody revealed three major patterns (Acrosomal-AR, apical-AP and, acrosome and tail-AT). The proportion of ASA reactive spermatozoa was significantly (P < 0.05) higher in low- and medium-fertile bulls compared to high-fertile bulls. Among the three patterns, the proportion of spermatozoa with AR pattern was significantly (P < 0.05) higher in low- and medium-fertile bulls compared to high-fertile bulls. The proportion of membrane and acrosome intact spermatozoa was significantly (P < 0.05) higher in high-fertile bulls compared to medium- and low-fertile bulls. There were no significant differences in the proportion of cryo-capacitated spermatozoa among high-, medium- and low-fertile bulls. The relationship between ASA reactive spermatozoa and conception rates (CR) of bulls was highly (P < 0.01) significant and negative. Similarly, AR and AT pattern were also significantly (P < 0.01) and negatively related to CR of bulls. The reactivity of spermatozoa with ASA was also significantly (P < 0.01) and negatively related to the membrane and acrosome integrity of spermatozoa. It was concluded that the proportion of spermatozoa responding to ASA was higher in low-compared to high-fertile bulls and ASA localization in sperm acrosomal area was negatively related to sperm membrane and acrosomal integrity and bull fertility. • Immunolocalization of sperm antigens was carried out using antisperm antibodies (ASA) in crossbred males. • The relationship between ASA reactive spermatozoa and bull conception rates was highly significant and negative. • The reactivity of spermatozoa with ASA was negatively related to membrane and acrosome integrity of spermatozoa. • Concluded that ASA binding to acrosome was negatively related to sperm membrane and acrosomal integrity, and bull fertility. [ABSTRACT FROM AUTHOR]

The effects of dietary supplementation with phytogenic blend (PB) of Aerva lanata, Piper betle, Cynodon dactylon , and Piper nigrum on growth performance, ileal nutrient digestibility, intestinal morphology, and cecal microflora were determined in a 42-day broiler feeding trial. A total of 192 broilers were assigned to 4 dietary treatments (6 replicates and 8 birds/replicate): basal diet, basal diet supplemented with antibiotic (chlortetracycline), 1% and 2% PB, respectively. The body weight gain (BWG) of starter chicks increased linearly (P = 0.023) as dietary supplementation levels of PB increased. At grower phase, broilers fed diet supplemented with 1% PB had similar BWG with the antibiotic group, but other treatments had reduced (P = 0.0001) BWG. Dietary supplementation with 1% PB resulted in the highest (P < 0.0001) BWG during the study. Feed intake was not affected by the treatments during the starter, finisher, and overall rearing periods. Broilers fed diet supplemented with 1% PB had the best (P < 0.0001) feed conversion ratio during the study. Overall, broilers fed only basal diet had the highest (P = 0.0450) mortality. Ileal organic matter (OM) digestibility increased linearly (P = 0.044) with broilers fed diet supplemented with PB, but reduced with antibiotic group. Dietary supplementation with 1% PB had the highest (P = 0.0402) ileal digestibility of tryptophan. In the duodenum, broilers fed diet supplemented with PB had longer (P = 0.0006) villi heights than the birds fed only basal diet, but similar with antibiotic group. Broilers fed diet supplemented with PB had longer (P = 0.0064) villi height in the jejunum than the antibiotic group. Bifidobacterium concentration of the cecum content showed a slight increase (P = 0.053) with increasing supplementation levels of PB. In conclusion, the current study shows that dietary supplementation with PB improves growth performance, intestinal morphology, and apparent ileal digestibility of OM and tryptophan in a dose-dependent manner with the best response at 1% inclusion level. [ABSTRACT FROM AUTHOR]

The present study aimed to isolate and enrich putative SSCs from ram testes, which are positive for promyelocytic leukaemia zinc-finger protein (PLZF). The putative SSCs were isolated using a combination of enzymes with different concentrations, collagenase (1 and 2 mg/ml), hyaluronidase (1 mg/ml) and trypsin (0.25 and 0.5 mg/ml). The isolated SSCs were purified using an extracellular matrix such as laminin (20 μg/ml), DSA-lectin (5 μg/ml) and gelatin (0.2%) in combination with BSA (0.5 mg/ml). The number of putative SSCs/ tubule was significantly ( p  < 0.05) higher in prepubertal (3.1 ± 0.51) and adult (3.45 ± 0.58) than the number of gonocytes/tubule in neonatal (0.59 ± 0.03) testis. Optimum enzyme combinations required for isolation of putative SSCs from prepubertal testis (collagenase; 2 mg/ml and trypsin; 0.5 mg/ml) were different from adult testis (collagenase; 1 mg/ml, trypsin; 0.25 mg/ml and hyaluronidase; 1 mg/ml). Though the number of putative SSCs/tubule was comparable in prepubertal and adult animals, a significantly ( p  < 0.05) higher percentage of putative SSCs (7.33 Vs 0.47%) were isolated from prepubertal testis than the adult. Differential plating using laminin along with BSA resulted in a significantly ( p  < 0.05) higher number of putative SSCs. The enzyme combinations suitable for isolation of putative SSCs from prepubertal testis are different from adult ram testis and the laminin has been found to be effective for purification of putative SSCs from testicular cells isolates. [ABSTRACT FROM AUTHOR]

Fertilization is an intricate cascade of events that irreversibly alter the participating male and female gamete and ultimately lead to the union of paternal and maternal genomes in the zygote. Fertilization starts with sperm capacitation within the oviductal sperm reservoir, followed by gamete recognition, sperm–zona pellucida interactions and sperm–oolemma adhesion and fusion, followed by sperm incorporation, oocyte activation, pronuclear development and embryo cleavage. At fertilization, bull spermatozoon loses its acrosome and plasma membrane components and contributes chromosomes, centriole, perinuclear theca proteins and regulatory RNAs to the zygote. While also incorporated in oocyte cytoplasm, structures of the sperm tail, including mitochondrial sheath, axoneme, fibrous sheath and outer dense fibers are degraded and recycled. The ability of some of these sperm contributed components to give rise to functional zygotic structures and properly induce embryonic development may vary between bulls, bearing on their reproductive performance, and on the fitness, health, fertility and production traits of their offspring. Proper functioning, recycling and remodeling of gamete structures at fertilization is aided by the ubiquitin–proteasome system (UPS), the universal substrate-specific protein recycling pathway present in bovine and other mammalian oocytes and spermatozoa. This review is focused on the aspects of UPS relevant to bovine fertilization and bull fertility. [ABSTRACT FROM AUTHOR]

The ability to predict the fertility of bulls before semen is released into the field has been a long-term objective of the animal breeding industry. However, the recent shift in the dairy industry towards the intensive use of young genomically selected bulls has increased its urgency. Such bulls, which are often in the highest demand, are frequently only used intensively for one season and consequently there is limited time to track their field fertility. A more pressing issue is that they produce fewer sperm per ejaculate than mature bulls and therefore there is a need to reduce the sperm number per straw to the minimum required without a concomitant reduction in fertility. However, as individual bulls vary in the minimum number of sperm required to achieve their maximum fertility, this cannot be currently achieved without extensive field-testing. Although an in vitro semen quality test, or combination of tests, which can accurately and consistently determine a bull's fertility and the optimum sperm number required represent the 'holy grail' in terms of semen assessment, this has not been achieved to date. Understanding the underlying causes of variation in bull fertility is a key prerequisite to achieving this goal. In this review, we consider the reliability of sire conception rate estimates and then consider where along the pregnancy establishment axis the variation in reproductive loss between bulls occurs. We discuss the aetiology of these deficiencies in sperm function and propose avenues for future investigation. [ABSTRACT FROM AUTHOR]

Spermatozoal transcripts expression levels could be used to assess fertility potential of a male. The objective of the present study was to elucidate the predictive ability of the expression levels of growth, apoptosis and homeostasis regulating transcripts on sperm functions and fertility. The expression levels of spermatozoal RNA isolated from the neat semen samples were related to the good (discarded ejaculate, <25%; n = 7) and poor (discarded ejaculate, >40%, n = 6) quality semen producer and bulls ( n = 12) with known conception rate. The relative fold expression levels of BMP2 were significantly ( p < 0.01) higher in good than the poor semen producers and positively associated with post-thaw sperm velocity parameters (LIN and VAP). The NGF expressions fold levels had significant ( p < 0.05) positive relationship with mitochondrial membrane potential of neat semen samples. The genes involved in the apoptotic, UBE2D3 ( r = −0.61, p = 0.02), CASP3 ( r = −0.57, p = 0.03) and homeostatic, HSFY2 ( r = −0.61, p < 0.02) regulators had significant negative correlation with the percentage of post-thaw fast progressive motile spermatozoa. The expression level of TRADD had significant negative influence on the mitochondrial membrane potential ( r = −0.54, p = 0.05) of neat semen samples and conception rate ( r = −0.57, p < 0.05). The expression levels of BMP2 had highly significant positive correlation with NGF ( r = 0.99, p < 0.01) and CASP3 ( r = 0.56, p = 0.05). The BMP2 expression level might be used to predict the quality of the semen and TRADD determine the conception rate of the bull. The study provides ample evidence that the sperm transcripts expression levels might be used to predict quality semen production and bull fertility. [ABSTRACT FROM AUTHOR]

The commercial applicability of bovine artificial insemination (AI) depends on the effectiveness of diluents for maintaining sperm fertility. Challenges faced by the AI industry due to recent advances in assisted reproduction, and the limitations inherent in using fresh and frozen-thawed sperm for AI, could be overcome with the development of better semen diluents. Research into the different microenvironments of bovine sperm as they progress towards maturity, capacitation and fertilisation is revealing various mechanisms that could be exploited to improve the formulation of semen diluents. These are reviewed here. A rationale for a more detailed investigation of bovine cervical mucus for factors that may allow further progress towards this goal are also discussed. [ABSTRACT FROM PUBLISHER]

Abstract: Although the existence of a complex population of mRNA in sperm is well documented, its role has not been completely elucidated. The objective of this study was to determine the relationship of mRNA abundance of sperm specific proteins and sire conception rates (SCR; a fertility index) in Holstein bulls. Samples of sperm from a single collection from commercial Holstein bulls (N = 34) were used to evaluate relative mRNA expression of adenylate kinase (AK) 1, integrin beta (IB) 5, Doppel, nerve growth factor, tissue inhibitors of metalloproteinases (TIMP) 2, lactate dehydrogenase C 1, small nuclear ribonucleoprotein polypeptide N, outer dense fiber 2, and phospholipase C zeta (PLCz) 1 in sperm. With the exception of lactate dehydrogenase C 1 and outer dense fiber 2, the mRNA abundances of these proteins were greater (P < 0.05) for high fertility (> +2 to ≤ 4 SCR) bulls compared with average (≥ 2 to ≤ +2) and low fertility (> −2 to ≤ −4) bulls. Of all the multivariate regression models tested, a combination of AK1, IB5, TIMP2, small nuclear ribonucleoprotein polypeptide N, and PLCz1 accounted for 97.4% of the variance in SCR scores. In the absence of PLCz1, the combination of AK1, IB5, Doppel, nerve growth factor, TIMP, and small nuclear ribonucleoprotein polypeptide N accounted for 96.6% of the variance in SCR scores. In addition, immunocytochemistry confirmed that the sperm-specific protein markers evaluated in this study were present in sperm. In conclusion, frozen-thawed semen from bulls with higher AK1, IB5, TIMP, small nuclear ribonucleoprotein polypeptide N 2 and PLCz1 mRNA abundances in the sperm had greater correlations with sire fertility index and may possess greater probabilities of siring calves. [Copyright &y& Elsevier]