Crossbred Bull Semen Research Articles

Time and dose-dependent effect of preconditioning with sodium nitroprusside (SNP) and 3-morpholinosydnonimine (SIN-1) on post-thaw semen quality of Karan-Fries (KF) bulls.

A novel strategy, focused on the induction of sub-lethal oxidative stress to optimize sperm cryosurvival, has been used before cryopreservation. The present study compared the effect of preconditioning with various concentrations of nitric oxide-donor (sodium nitroprusside, SNP) and peroxynitrite-generator (3-morpholinosydnonimine, SIN-1) on in vitro sperm functions and lipid peroxidation status (LPO) of cryopreserved Karan-Fries (KF) crossbred bull semen. To optimize the concentration of additives, spermatozoa obtained from 36 ejaculates were supplemented with different concentrations of SNP (0.01, 0.05, 0.1μM) and SIN-1 (80, 160, 200μM) versus control in the extender. The post-freezing sperm motility and viability were greater (p < 0.05) in 0.1μM SNP and 80μM SIN-1 in comparison to other concentrations used. Furthermore, the spermatozoa obtained from 48 ejaculates were supplemented with 0.1μM SNP and 80μM SIN-1 in the extender. A significant increase (p < 0.05) was observed in progressive motility, viability and membrane integrity in SNP and SIN-1 treated extender at 24h, 15days, and 2-month post-cryopreservation (PC) periods. There was no significant difference in sperm abnormality in the extended groups and the control group. The seminal plasma of SNP-treated extender had less (p < 0.05) lipid peroxidation as compared to SIN-1 treated and control groups. In post-thaw semen, both SNP and SIN-1 showed a higher (p < 0.05) proportion of acrosome intact (FITC-PNA) sperm with a greater decrease (p < 0.05) in membrane scrambling and lipid peroxidation. SNP and SIN-1 improved (p < 0.05) the proportion of sperm with higher mitochondrial membrane potential (Δψm) as compared to the control. In conclusion, it seems that the preconditioning of SNP and SIN-1 at lower doses may have beneficial effects on post-thawed crossbred bull sperm quality.

Optimization of dissolved oxygen levels in extender prevents development of cryocapacitation like changes, oxidative stress and augments zona binding capacity of crossbred bull spermatozoa.

The present study was undertaken to determine the efficacy of partial deoxygenation of extender at constant temperature (35°C) in freezability of crossbred bull semen. The dissolved oxygen (DO) levels were reduced by the use of newly developed technique of nitrogen effervescence at a flow rate of 2-3 bubbles per second. Four different levels of oxygen in semen extender, that is 11.7, 2, 4 and 8ppm as control (Group-I), Group-II, Group-III and Group-IV, respectively, were used to assess the effect of partial deoxygenation on semen quality parameters. The 4ppm level of DO resulted in higher (p<0.05) progressive motility in comparison with non-treated group at post-thaw stage, whereas reduction up to 2ppm resulted in drastic fall in motility. Oxidative stress status revealed low superoxide dismutase (SOD) and total antioxidant capacity (TAC) in Group-II, whereas higher (p<0.05) SOD and TAC activities were observed in Group-III in comparison with non-treated group at pre-freeze and post-thaw stages. The sperm-zona binding at 4ppm level of DO was significantly higher than control group, 2 and 8ppm levels of DO. In conclusion, reduction of DO in the extender up to 4ppm reduced oxidative stress and improved in vitro fertility of crossbred bull spermatozoa.

Cellular and Molecular Insights Into the Etiology of Subfertility/Infertility in Crossbred Bulls (Bos taurus × Bos indicus): A Review.

Crossbreeding of indigenous cattle (Bos indicus) with improved (Bos taurus) breeds gained momentum and economic relevance in several countries to increase milk production. While production performance of the crossbred offspring is high due to hybrid vigor, they suffer from a high incidence of reproductive problems. Specifically, the crossbred males suffer from serious forms of subfertility/infertility, which can have a significant effect because semen from a single male is used to breed several thousand females. During the last two decades, attempts have been made to understand the probable reasons for infertility in crossbred bulls. Published evidence indicates that testicular cytology indices, hormonal concentrations, sperm phenotypic characteristics and seminal plasma composition were altered in crossbred compared to purebred males. A few recent studies compared crossbred bull semen with purebred bull semen using genomics, transcriptomics, proteomics and metabolomics; molecules potentially associated with subfertility/infertility in crossbred bulls were identified. Nevertheless, the precise reason behind the poor quality of semen and high incidence of sub-fertility/infertility in crossbred bulls are not yet well defined. To identify the underlying etiology for infertility in crossbred bulls, a thorough understanding of the magnitude of the problem and an overview of the prior art is needed; however, such systematically reviewed information is not available. Therefore, the primary focus of this review is to compile and analyze earlier findings on crossbred bull fertility/infertility. In addition, the differences between purebred and crossbred males in terms of testicular composition, sperm phenotypic characteristics, molecular composition, environmental influence and other details are described; future prospects for research on crossbred males are also outlined.

Differentially expressed microRNAs in biochemically characterized FrieswalTM crossbred bull semen

In addition to the transmission of paternal genome, spermatozoa also carry coding as well as noncoding microRNAs (miRNAs) into the female oocyte during the process of biological fertilization. Based on RNA deep sequencing, a total 28 number of differentially expressed miRNAs were cataloged in categorized FrieswalTM crossbred (Holstein Friesian X Sahiwal) bull semen on the basis of conception rate (CR) in field progeny testing program. Validation of selected miRNAs viz. bta-mir-182, bta-let-7b, bta-mir-34c and bta-mir-20a revealed that, superior bull semen having comparatively (p < .05) lower level of all the miRNAs in contrast to inferior bull semen. Additionally, it was illustrated that, bta-mir-20a and bta-mir-34c miRNAs are negatively (p < .01) correlated with seminal plasma catalase (CAT) activity and glutathione peroxidase (GPx) level. Interactome studies identified that bta-mir-140, bta-mir-342, bta-mir-1306 and bta-mir-217 can target few of the important solute carrier (SLC) proteins viz. SLC30A3, SLC39A9, SLC31A1 and SLC38A2, respectively. Interestingly, it was noticed that all the SLCs were significantly (p < .05) expressed at higher level in superior quality bull semen and they are negatively correlated (p < .01) with their corresponding miRNAs as mentioned. This study may reflect the role of miRNAs in regulating few of the candidate genes and thus may influence the bull semen quality traits.

In vitro Fertilisation Capacity of Frozen Crossbred Bull Semen Cryopreserved During Different Seasons in Kerala

Background: Assessment of semen quality of bulls in frozen semen stations is of paramount importance as they are used for inseminating large number of cattle. The present study was conducted to assess the in vitro fertilisation capacity of crossbred bull semen cryopreserved during different seasons in Kerala as reports of such an evaluation are scarce.Methods: Semen samples from six crossbred bulls of same exotic inheritance, cryopreserved during rainy, post monsoon and summer seasons were procured from KLDB, Dhoni. The collected samples were evaluated for their in vitro fertilisation potential.Result: Hot dry summer season in Kerala adversely affects the fertilisation capacity of spermatozoa. Rainy season was observed to be the most favourable season for good quality semen production and post monsoon season was intermediate between summer and rainy season. It can be concluded from the present investigation that semen cryopreserved during summer season have lower fertilisation and cleavage rate than rainy and post monsoon season. This might be due to the harmful effect of significantly higher average maximum temperature and lower relative humidity occurred in the area during summer season on spermatogenesis.

Quality of frozen-thawed crossbred bull semen diluted with vitamin E and Tocopherol with omega III enriched egg yolk

Quality of frozen-thawed crossbred bull semen diluted with vitamin E and Tocopherol with omega III enriched egg yolk

Database on spermatozoa transcriptogram of catagorised Frieswal crossbred (Holstein Friesian X Sahiwal) bulls

Bull spermatozoa contain different functional genes and many of them plays important roles in different stages of spermatogenesis, spermatozoa kinetics, fertilization as well as embryonic development. RNA deep sequencing is one of the preferred tools for absolute quantification of messenger RNA. The intention of the current study was to investigate the abundance of spermatozoal transcripts in categorized Frieswal (Holstein-Friesian X Sahiwal) crossbred bull semen through RNA deep sequencing. A total 1546561 and 1019308 numbers of reads were identified among good and poor quality bull spermatozoa based on their conception rate. Post mapping with Bos taurus reference genome identified 1,321,236 and 842,022 number of transcripts among good and poor quality RNA libraries, respectively. However, a total number of 3510 and 6759 functional transcripts were identified among good and poor quality bull spermatozoa, respectively. Most of the identified transcripts were related to spermatozoa functions, embryonic development and other functional aspects of fertilization. Wet laboratory validation of the top five selected transcripts (AKAP4, PRM1, ATP2B4, TRIM71 and SLC9B2) illustrated the significant (p < 0.01) level of expression in the good quality crossbred bull semen than the poor quality counterparts. The present study with comprehensive profiling of spermatozoal transcripts provides a useful non-invasive tool to understand the causes of as well as an effective way to predict male infertility in crossbred bulls.

Functional proteomic analysis of crossbred (Holstein Friesian×Sahiwal) bull spermatozoa.

Male infertility is one of the prime concerns of dairy cattle production. The study was designed to find out differentially expressed proteins in categorized crossbred (Holstein Friesian×Sahiwal) bull semen to serve as potential biomarkers for male infertility. Frozen crossbred bull semen with satisfactory phenotypic records were defined as "good" and "poor" based on their fertility rates. A total of 1,547 proteins were detected in bull spermatozoa using liquid chromatography-mass spectrometer (LC-MS/MS) analysis. Results revealed that 558 (36.1%) and 653 (42.2%) proteins were expressed to good and poor quality bull spermatozoa, respectively. A total of 336 proteins (21.7%) were reported to be unique for both good and poor quality bull semen, and among the common proteins, 224 (66.7%) and 112 (33.3%) were up- and downregulated in good and poor quality categorized bull semen, respectively. Gene Ontology analysis of global proteomes identified different signalling pathways, and most of them were related to cellular motility, immune systems as well as cellular metabolisms. The distinctive presence of some of the proteins may provide an insight into the molecular mechanistic role played by these proteins in crossbred bull infertility.

Assessment of antioxidants for preservation of crossbred bull semen in Tris based extender

Present study was aimed to improve poor quality crossbred bulls semen, as occurrence of poor quality ejaculates are one of the major problems in crossbred bulls. Antioxidants and its combination was tried to overcome such problems. Vitamin E, Vitamin C and Vitamin E+C were supplemented at dose rate of 1mg, 5 mM and 1mg + 5mM per ml respectively in split ejaculates to evaluate semen quality during preservation at refrigerated temperature and cryopreservation. The results showed that semen characteristics were better in antioxidant supplemented ejaculates. Total sperm abnormality were significantly lower (p&gt;0.05) in antioxidant supplemented group. The seminal characteristics of crossbred bull semen showed significantly better (p&gt;0.05) performance on preservation, when fortified with Vitamin E as compared to Vitamin C alone as well as Vitamin E and C in combination. The performance of semen additives, were supportive to semen characteristics especially for Vitamin E, moreover fortification with Vitamin E+C was slightly superior to Vitamin C. It can be concluded that fortification of Vitamin E has beneficial role in semen quality improvement followed by Vitamin E+C and Vitamin C.

Assessment of antioxidants for preservation of crossbred bull semen in Tris based extender

Present study was aimed to improve poor quality crossbred bulls semen, as occurrence of poor quality ejaculates are one of the major problems in crossbred bulls. Antioxidants and its combination was tried to overcome such problems. Vitamin E, Vitamin C and Vitamin E+C were supplemented at dose rate of 1mg, 5 mM and 1mg + 5mM per ml respectively in split ejaculates to evaluate semen quality during preservation at refrigerated temperature and cryopreservation. The results showed that semen characteristics were better in antioxidant supplemented ejaculates. Total sperm abnormality were significantly lower (p<0.05) in antioxidant supplemented group. The seminal characteristics of crossbred bull semen showed significantly better (p<0.05) performance on preservation, when fortified with Vitamin E as compared to Vitamin C alone as well as Vitamin E and C in combination. The performance of semen additives, were supportive to semen characteristics especially for Vitamin E, moreover fortification with Vitamin E+C was slightly superior to Vitamin C. It can be concluded that fortification of Vitamin E has beneficial role in semen quality improvement followed by Vitamin E+C and Vitamin C.

Bovine Circadian Locomotor Output Cycles Kaput (CLOCK) and Clusterin (CLU) mRNA Quantitation in Ejaculated Crossbred Bull Spermatozoa.

Mammalian circadian locomotor output cycles kaput (CLOCK) gene encodes a transcription factor that affects both the persistence and the period of circadian rhythms. Earlier reports suggested that CLOCK gene might be associated with male infertility in human. Present investigation, for the first time, reports that CLOCK gene expresses differentially between good and poor quality crossbred bull semen. The relative expression of CLOCK was significantly (p<0.05) higher among good quality bull semen than motility-impaired ones. Clusterins (CLU) are series of genes associated with a variety of physiological activities including spermatogenesis, apoptosis and degenerative disease conditions. In the present context, we also investigated that the expression of CLU gene was significantly (p<0.05) higher among motility-impaired crossbred bull semen compared to the good quality one.

Designing of an artificial neural network model to evaluate the association of three combined Y-specific microsatellite loci on the actual and predicted postthaw motility in crossbred bull semen

The freezing of bull semen significantly hamper the motility of sperm which reduces the conception rate in dairy cattle. The prediction of postthaw motility (PTM) before freezing will be useful to take the decision on discarding or freezing of the germplasm. The artificial neural network (ANN) methodology found to be useful in prediction and classification problems related to animal science, and hence, the present study was undertaken to compare the efficiency of ANN in prediction of PTM on the basis of the number of ejaculates, volume, and concentration of sperms. The combined effect of Y-specific microsatellite alleles on the actual and predicted PTM was also studied. The results revealed that the prediction accuracy of PTM based on the semen quality parameters was comparatively lower because of higher variability in the data set. The ANN gave better prediction accuracy (34.88%) than the multiple regression analysis models (32.04%). The root mean square error was lower for ANN (8.4353) than that in the multiple regression analysis (8.6168). The haplotype or combined effect of microsatellite alleles on actual and predicted PTM was found to be highly significant (P < 0.01). On the basis of results, it was concluded that the ANN methodology can be used for prediction of PTM in crossbred bulls.

Reduced glutathione and cysteine hydrochloride on crossbred bull semen

Reduced glutathione and cysteine hydrochloride on crossbred bull semen

Reduced glutathione and cysteine hydrochloride on crossbred bull semen

AbstractOver the last few years, good quality freezable semen was not utilized effectively due to less knowledge about the freezability of semen. One reason is the lack of sufficient antioxidants in the seminal plasma and semen extender. The antioxidant content is reduced during the cryopreservation process and causes premature cryo-capacitation and modification of sperm membrane structure upon thawing. So the study of pre-freezing and post thaw seminal parameters like viability, motility, acrosomal integrity, lipid peroxide assay, vanguard distance traveled by sperm, mitochondrial membrane potential, velocity and motility parameters (CASA) and field fertility trial helped to assess the effectiveness of the additives. The conception rate was better in reduced glutathione than the cysteine hydrochloride treated group. The glutathione has improved the poor freezing and maintained the good freezable semen. Thus the field fertility rate was enhanced and in turn helped to prevent waste of good quality germplasm and repeat breeder syndrome in cows.