Evaluation of predictive ability of linear type gonadal traits on reproductive capacity of breeding dairy bulls.

Dimensions of linear type traits facilitate selection of livestock for breeding and rearing. To date, use of linear type traits for selection of breeding bulls is highly concentric to scrotal circumference (SC), with probable overlook to other important traits. Present study reported the importance of various gonadal linear type traits on spermatozoa production, age-related changes in gonadal linear type traits of bulls and predictive ability of these traits on bulls' reproductive potentials. Among all gonadal traits, testicular density (TD), scrotal volume (SV), paired testicular weight (PWT) and SC were found most important predictor variables in order, which can discriminate between good/poor breeding bulls, that is, produced frozen semen doses (FSD) or not. Dimensions of gonadal traits increased significantly up to 36 months age and thereafter, development became slow and negligible. In contrast, TD decreased by 30%, 51%, 64%, 68% and 71% at 12, 24, 36, 48 and >49 months age, respectively, from its base value at 6 months. Bulls of lower TD (≤0.88 g/cm3) had significantly higher ejaculate volume (+9%), sperm motility, sperm concentration (+100 million/mL) and sperm output (+26%)/ejaculate as compared to bulls of higher TD (>0.88 g/cm3). Discriminant function was developed using TD, SV, PWT and SC to identify bulls of superior reproductive potentials. It was concluded that among the investigated traits, TD was the strongest to discriminate between FSD and Non-FSD bulls. Therefore, our findings suggested that TD could be more potential trait than SC for dairy bulls' breeding soundness evaluation and assessment of reproductive ability.

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.

Effect of seasons and photoperiods on seminal attributes and sperm morphology in Holstein Friesian × Sahiwal crossbred dairy bulls.

Cattle being non-seasonal breeding species, effects of photoperiods on sperm traits and morphology had been reported inadequately in breeding bulls. To elucidate the plausible existence of effects of photoperiods and seasons, seminal traits of Holstein Friesian × Sahiwal crossbred dairy bulls (N = 557) were analysed using different statistical models. A present study revealed that the biological rhythm of reproduction oscillated almost in parallel to the annual changes of natural photoperiods even in non-seasonal breeding species like cattle bulls. Semen traits diminished to the lowest in winter solstice (WS ± 45 days), progressively increased with the rising day length of spring (vernal equinox ± 45 days), reached a peak in summer solstice (SS ± 45 days), and then gradually reduced with decreasing photoperiod of the autumn equinox (AE ± 45 days). From summer solstice to winter solstice, sperm concentration reduced by 90.53 million/ml (8.85%), total sperm count/ejaculate decreased by 785 million (13.87%), total motile sperm count/ejaculate reduced by 17.59%, and total post-thaw motile sperm counts/ejaculates diminished by 38.64%. In short-duration photoperiods (≤ 12 h), bulls had a significantly (P < 0.01) higher incidence of major, minor, tail abnormality and total aberrant sperm% compared to that of long-duration photoperiods (> 12 h). Solstice equinox-based seasonal classification provided better insight into photoperiodicity on bulls' semen quality and sperm traits as compared to conventional meteorological classification of seasons. It was concluded that photoperiods affect sperm productivity, semen quality, and sperm morphology in non-seasonal breeding species like dairy bulls, maintained at transitional latitude (29° N) tropical climate. Bulls' reproductive ability was more influenced by the phases of increasing/decreasing day length duly primed by climax/trough of photoperiods, compared to mere long/short duration of photoperiods.

Association of linear type pelvic traits with testicular characteristics and reproductive capability of breeding dairy bulls.

Associations of pelvic linear type traits (PLT) on production and reproduction of cows were widely documented; in contrast, importance of PLT on bulls' reproductive ability, semen quality, semen cryo-preservability, frozen semen doses (FSD) production etc. was inadequately reported. The present study was conducted on Frieswal bulls (N = 378, age: 6-91 months, m) to assess age-wise growth dynamics of pelvic dimensions, classifying age into 6 groups (6 m, 7-12 m, 13-24 m, 25-36 m, 37-48 m, >48 m). Iso-age group bulls' (N = 100; 25-36 m) records were analysed after classifying seasons (summer/rainy/winter) of semen collection, if pelvic morphometric traits had any practical associations with testicular traits, semen quality and FSD production or not. A discriminant function was developed based on PLT, which showed poor predictive ability to distinguish FSD/non-FSD category bulls. Age significantly influenced growth and dimensions of gonadal and external pelvic morphometry traits. The dimensions of PLT increased at higher rate up to 36 m age and thereafter enhancement became insignificant. Pelvic linear type traits were positively (p < .01) associated with testicular/scrotal traits. Among PLT studied, pelvic triangle area (PTA) was the strongest discriminating variable to distinguish between good and poor breeding bulls. Bulls of larger PTA (≥1000 cm2 ), at average 30 m age, produced relatively inferior-quality semen, viz. lower volume (-3%), sperm concentration (-48 × 106 /mL), motility, semen quality index, total sperm counts/ejaculate (-601 × 106 ), motile sperm counts/ejaculate (-474 × 106 ) and total post-thaw motile sperm counts (-226 106 ), than bulls of lesser PTA (<1000 cm2 ). It was concluded that external PTA could be a useful addition to the bulls' breeding soundness evaluation.

Non-gonadal linear type traits can discriminate reproductive ability of breeding dairy bulls.

Linear type traits are easily measurable phenotypic characteristics that help breed characterization, selection of animals for breeding and found to be associated with animals' performance. Unlike cows, there have been limited studies linking body linear traits with male reproductive ability and semen cryo-preservability of breeding bulls. Present study reported the age-related changes in body linear type traits in Frieswal (N = 378) dairy bulls and its relevance with reproductive potentials of breeding bulls. Our results indicated that body frame size traits were significantly and positively correlated with gonadal linear traits. Among the selected body mophometric parameters body length, chest girth and head circumference were the important body linear type traits having capability to discriminate between bulls of frozen semen doses (FSD) and non-FSD categories. Discriminant function has been developed based on body linear traits of crossbred dairy bulls to find out males of superior reproductive potentials. Our finding provided evidence that body length (humerous tuberosity to tuber ischii) was the most powerful linear body trait associated with breeding bulls' reproductive ability and semen quality.

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.

Molecular markers, BM1500 and UMN2008, are associated with post-thaw motility of bull sperm.

Cryopreservation is one of the most important aspects of frozen semen technology and livestock breeding. Uses of candidate molecular markers in selection strategies for male fertility are well recognized. The present investigation targeted two microsatellite markers (BM1500 and UMN 2008) for association with semen quality variables and freezing capacity in Frieswal (HF×Sahiwal) crossbred bulls of Indian origin. Of the different alleles at the polymorphic locus BM1500, the 136bp allele was associated with greater (P<0.05) post-thaw motility percentage (PTM) while the 134 allele was associated with less (P<0.05) PTM. The 134/134 genotype at the polymorphic locus, UMN2008 was associated with greater (P<0.05) post-thaw motility while there was no allele effect on PTM. When combined genotypes UMN2008/BM1500 were analyzed, the 134/134-136/136genotype had the greatest (P<0.05) association with PTM. The present study is an initial report on the potential use of these markers as male reproductive biomarkers for improving semen freezing capacity in bulls.

A SNP (g.358A > T) at intronic region of CD9 molecule of crossbred bulls may associate with spermatozoal motility.

The surface expression of CD9 (cluster-of-differentiation antigen-9) in sperms of certain mammalian species has been attributed to its fusion with the egg and thereby dictating the fertility of species. In the present study, we investigated the association of CD9 with crossbred bull sperm quality and quantity trait was analyzed using a total of 96 Frieswal (HF × Sahiwal) crossbred. A single nucleotide polymorphism (g.358A > T) in intron 6 was significantly associated with sperm concentration (P < 0.05) and motility percentage (P < 0.01). mRNA was extracted from good (progressive motility > 50%) and motility impaired (progressive motility < 50%) bull semen. The mRNA expression and seminal plasma protein concentration of CD9 was significantly (P < 0.05) higher among good quality bull semen than motility impaired ones. Our results thus may indicate that, mutation in the intronic region may be responsible for the instability of RNA and the subsequent functional protein expression.

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.