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Seminal plasma proteins and pathways associated with sperm motility have not been elucidated in stallions. Therefore, in the current study, using the high throughput LC/MS-MS approach, we profiled stallion seminal plasma proteins and identified the proteins and pathways associated with sperm motility. Seminal plasma from six stallions producing semen with contrasting sperm motility (n = 3 each high-and low-motile group) was utilized for proteomic analysis. We identified a total of 1687 proteins in stallion seminal plasma, of which 1627 and 1496 proteins were expressed in high- (HM) and low- motile (LM) sperm of stallions, respectively. A total number of 1436 proteins were co-expressed in both the groups; 191 (11%) and 60 (3.5%) proteins were exclusively detected in HM and LM groups, respectively. A total of 220 proteins were upregulated (>1-fold change) and 386 proteins were downregulated in SP from LM group stallions as compared to HM group stallions, while 830 proteins were neutrally expressed in both the groups. Gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed dysregulation of the important proteins related to mitochondrial function, acrosome, and sperm cytoskeleton in the seminal plasma of stallions producing ejaculates with low sperm motility. High abundance of peroxiredoxins and low abundance of seminal Chaperonin Containing TCP1 Complex (CCT) complex and Annexins indicate dysregulated oxidative metabolism, which might be the underlying etiology for poor sperm motility in LM group stallions. In conclusion, the current study identified the seminal plasma proteomic alterations associated with poor sperm motility in stallions; the results indicate that poor sperm motility in stallions could be associated with altered expression of seminal plasma proteins involved in oxidative metabolism. [ABSTRACT FROM AUTHOR]

Bull fertility is of paramount importance in bovine industry because semen from a single bull is used to breed several thousands of cows; however, so far, no reliable test is available for bull fertility prediction. In the present study, spermatozoa from high- and low-fertility bulls were subjected to high-throughput transcriptomic, proteomic and metabolomic analysis. Using an integrated multi-omics approach the molecular differences between high- and low-fertility bulls were identified. We identified a total of 18,068 transcripts, 5041 proteins and 3704 metabolites in bull spermatozoa, of which the expression of 4766 transcripts, 785 proteins and 33 metabolites were dysregulated between high- and low-fertility bulls. At transcript level, several genes involved in oxidative phosphorylation pathway were found to be downregulated, while at protein level genes involved in metabolic pathways were significantly downregulated in low-fertility bulls. We found that metabolites involved in Taurine and hypotaurine metabolism were significantly downregulated in low-fertility bulls. Integrated multi-omics analysis revealed the interaction of dysregulated transcripts, proteins and metabolites in major metabolic pathways, including Butanoate metabolism, Glycolysis and gluconeogenesis, Methionine and cysteine metabolism, Phosphatidyl inositol phosphate, pyrimidine metabolism and saturated fatty acid beta oxidation. These findings collectively indicate that molecules governing sperm metabolism potentially influence bull fertility. [ABSTRACT FROM AUTHOR]

The article discusses the significance of equine conservation in India and highlights the successful implementation of embryo transfer technology to preserve indigenous horse breeds, exemplified by the birth of Raj-Prathama, India's first Marwari filly born through this method. Topics include the declining population of equines, the role of the National Research Centre on Equines in genetic conservation, and the potential of embryo transfer technology to enhance breed preservation efforts.

During transit and storage in epididymis, spermatozoa undergo final maturation, acquire motility, functional competence and the ability to fertilise an oocyte. Epididymal secretions contain a complex biochemical milieu of diverse inorganic ions, proteins, metabolites and other molecules. Since it is believed that spermatozoa are translationally silent, proteins appearing in them are thought to be synthesised elsewhere, including epididymis, and then incorporated to the cells. One of the important mechanisms suggested to be involved in transfer of epididymal secretions to spermatozoa is through exosomes called epididymosomes. Epididymosomes released from the epididymal epithelium contain proteins, noncoding RNAs and distinct set of lipids that are transferred to spermatozoa while they pass through the different epididymal regions. Owing to the importance of these molecules for sperm maturation and fertilising ability, research on epididymosomes has gained increasing attention during the last decade. This review is focused on epididymosomes, with emphasis on recent advances in the understanding of mechanisms of epididymosomal cargo transfer to spermatozoa and potential roles of epididymosomes in sperm function and beyond. Possibilities of utilising the molecular signatures of epididymosomes as a tool for male fertility assessment are also discussed. [ABSTRACT FROM AUTHOR]

Aim: To investigate various biochemical components of seminal plasma in Marwari stallions and Poitou Jacks and to find out their correlation with that of the seminal characteristics. Materials and Methods: In this study, semen was collected from six Marwari stallions and six Poitou jacks aged from 4 to 6 years and with known fertility status. The semen collection from the stallions were collected during the breeding season, i.e., between the months of April and June. From the collected semen ejaculates, we estimated the values of some biochemical components, viz., total protein content, total lipid content, and enzymes such as glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT), alkaline phosphatase (ALP), acid phosphatase (ACP), and lactate dehydrogenase (LDH) as well as concentrations of glucose, cholesterol, total calcium (Ca), and phosphorus (P) and correlations among different seminal parameters were statistically examined using the Pearson correlation coefficient. Results: In this study, we found positive correlations between semen volume as well as sperm concentration and GOT, GPT, ALP and ACP for both the group stallions. Significant correlation between motility and glucose, GOT and GPT could be an indication for their role metabolism and protection against free radicals to the spermatozoa. Conclusion: Based on the results, it is concluded that there is a positive correlation between some biochemical values such as glucose, Ca, ALP, and LDH and seminal parameters which play a key role in capacitation and onward movement of the spermatozoa. [ABSTRACT FROM AUTHOR]

The reason for poor semen quality among the breeding bulls is not well understood. In the present study, we performed high‐throughput RNAseq analysis of spermatozoa to identify the SNPs present in good and poor‐quality semen‐producing Holstein Friesian breeding bulls. A total of 21,360 and 44,650 SNPs were identified in good and poor‐quality semen with a minimum read depth of 20, among which 4780 and 8710 novel variants were observed in good and poor‐quality semen, respectively. Greater SNPs and indels variations were observed in poor compared to good‐quality semen. In poor‐quality semen, SNP variations were observed in ZNF280B, SLC26A2, DMXL1, OR52A1, MACROD2 and REV1 genes, which are associated with regulation of spermatogenesis, post‐testicular maturation, Cl− channel activity, V‐ATPase‐mediated intracellular vesicle acidification, a mono‐ADP‐ribosyl hydrolase and ATR‐Chk1 checkpoint activation. GO analysis of filtered genes with significant variations between good and poor‐quality semen showed enrichment in important pathways related to semen quality such as MAPK signalling pathway, Akt signalling pathway, focal adhesion, cAMP signalling pathway, and Rap1 signalling pathway. Network analysis of filtered genes in poor‐quality semen showed variations in pathways of purine metabolism, pyrimidine metabolism, prolactin signalling pathway and RNA cap‐binding complex. It is inferred that SNP in genes involved in maintaining sperm functions could be the reason for poor‐quality semen production in bulls, and the identified SNPs hold potential to be used as biomarkers for semen quality in bulls. [ABSTRACT FROM AUTHOR]

Clinical diagnosis of pyometra in an Arabian mare was undertaken through transrectal ultrasonography and was managed with uterine lavage and intrauterine antibiotics. [ABSTRACT FROM AUTHOR]

The report records diagnosis and management of endometrial cysts in mares. The endometrial cysts were diagnosed through ultrasonography and their size was determined. As cysts were small in nature, they were treated and pregnancy was established. [ABSTRACT FROM AUTHOR]

The article presents a study which examined the presence of cryptorchidism in a Marwari stallion based on the biometry of scrotum and level of testosterone. The variables considerfed include the level of blood testosterone which is validated by electrochemiluminescence method, semen sampling through artificial vaginal method and sperm concentration. A comparison of testosterone level in monorchid stallion, normal Marwaei stallion and exotic Poitou jacks is offered.

Although, it is well understood that sperm DNA damage is associated with infertility, the molecular details of how damaged sperm DNA affects fertility are not fully elucidated. Since sperm proteins play an important role in fertilization and post-fertilization events, the present study aimed to identify the sperm proteomic alterations in bulls with high sperm DNA Fragmentation Index (DFI%). Semen from Holstein-Friesian crossbred breeding bulls (n = 50) was subjected to Sperm Chromatin Structure Assay. Based on DFI%, bulls were classified into either high- (HDFI; n = 6), or low-DFI (LDFI; n = 6) and their spermatozoa were subjected to high throughput proteomic analysis. Liquid chromatography and mass spectrometry analysis identified 4567 proteins in bull spermatozoa. A total of 2660 proteins were found common to both the groups, while 1193 and 714 proteins were unique to HDFI and LDFI group, respectively. A total of 265 proteins were up regulated and 262 proteins were down regulated in HDFI group. It was found that proteins involved in capacitation [heparin binding (molecular function), ERK1 and ERK2 cascade (biological process), PI3K-Akt signalling (pathway), Jak-STAT signalling (pathway)], spermatogenesis [TLR signalling (pathway), gamete generation (biological process)] and DNA repair mechanism (biological process) were significantly altered in the bulls with high DFI%. • Comparative sperm proteomic profile between bulls with high- (HDFI) and low-DNA Fragmentation Index (LDFI) was studied. • Identified 1193 and 714 unique proteins in HDFI and LDFI bulls, respectively. • A total of 265 and 262 proteins were up- and down-regulated in HDFI bulls. • Proteins involved in DNA repair mechanism were significantly altered in HDFI bulls. • Expression of proteins involved in spermatogenesis and capacitation were dysregulated in HDFI bulls. [ABSTRACT FROM AUTHOR]

The horse, one of the most domesticated animals, has been used for several purposes, like transportation, hunting, in sport, or for agriculture-related works. Kathiawari, Marwari, Manipuri, Zanskari, Bhutia, Spiti, and Thoroughbred are the main breeds of horses, particularly due to their agroclimatic adaptation and role in any kind of strong physical activity, and these characteristics are majorly governed by genetic factors. The genetic diversity and phylogenetic relationship of these Indian equine breeds using microsatellite markers have been reported, but further studies exploring the SNP diversity and runs of homozygosity revealing the selection signature of breeds are still warranted. In our study, the identification of genes that play a vital role in muscle development is performed through SNP detection via the whole-genome sequencing approach. A total of 96 samples, categorized under seven breeds, and 620,721 SNPs were considered to ascertain the ROH patterns amongst all the seven breeds. Over 5444 ROH islands were mined, and the maximum number of ROHs was found to be present in Zanskari, while Thoroughbred was confined to the lowest number of ROHs. Gene enrichment of these ROH islands produced 6757 functional genes, with AGPAT1, CLEC4, and CFAP20 as important gene families. However, QTL annotation revealed that the maximum QTLs were associated with Wither's height trait ontology that falls under the growth trait in all seven breeds. An Equine SNP marker database (EqSNPDb) was developed to catalogue ROHs for all these equine breeds for the flexible and easy chromosome-wise retrieval of ROH along with the genotype details of all the SNPs. Such a study can reveal breed divergence in different climatic and ecological conditions. [ABSTRACT FROM AUTHOR]

Seminal plasma proteins are the major extrinsic factors that can modulate the sperm quality and functions. The present study was carried out to compare the proteomic profiles of seminal plasma from breeding bulls producing good and poor quality semen in an effort to understand the possible proteins associated with semen quality. A total of 910 and 715 proteins were detected in the seminal plasma of poor and good quality semen producing bulls, respectively. A total of 705 proteins were common to both the groups, in which 380 proteins were upregulated and 89 proteins were downregulated in the seminal plasma of poor quality semen, while 236 proteins were co-expressed. The proteins negatively influencing sperm functions such as CCL2, UQCRC2, and SAA1 were among the top ten upregulated proteins in the seminal plasma of poor quality semen. Proteins having a positive role in sperm functions (NGF, EEF1A2, COL1A2, IZUMO4, PRSS1, COL1A1, WFDC2) were among the top ten downregulated proteins in the seminal plasma of poor quality semen. The upregulation of oxidation–reduction process-related proteins, histone proteins (HIST3H2A, H2AFJ, H2AFZ, H2AFX, HIST2H2AB, H2AFV, HIST1H2AC, HIST2H2AC, LOC104975684, LOC524236, LOC614970, LOC529277), and ubiquinol–cytochrome-c reductase proteins (UQCRB, UQCRFS1, UQCRQ, UQCRC1, UQCRC2) indicate deranged oxidation–reduction equilibrium, chromatin condensation and spermatogenesis in poor quality semen producing bulls. The expression of proteins essential for motile cilium (CCDC114, CFAP206, TEKT4), chromatin integrity (PRM2), gamete fusion (IZUMO4, EQTN), hyperactivation, tyrosine phosphorylation, and capacitation [PI3K–Akt signalling pathway-related proteins (COL1A1, COL2A1, COL1A2, SPP1, PDGFA, NGF)] were down regulated in poor quality semen producing bulls. [ABSTRACT FROM AUTHOR]

Antioxidants are used to minimize oxidative stress during liquid semen storage. The main aim of current study was to elucidate effect of supplementing melatonin and canthaxanthin in Tris‐based extender could enhance seminal quality of ram at 4°C up to 72 h. A total of 48 ejaculates were collected from breeding Magra rams (n = 8) and were preliminarily subjected for various macroscopic and microscopic semen evaluation tests. These ejaculates were pooled and divided into three equal aliquots. Two aliquots were diluted (1:10) using extender encompassing final concentration of 1mM melatonin and 25 µM canthaxanthin and stored at 4°C. Third aliquot with extender only was kept as control. Structural and functional seminal changes were observed at different time points of preservation. Results revealed that mean values for progressive sperm motility, viability and total antioxidant capacity were significantly higher (p < 0.05) in melatonin group while hypo‐osmotic swelling test was significantly (p < 0.05) higher in canthaxanthin group. Total sperm abnormalities and malondialdehyde levels were significantly (p < 0.05) lower in both treatment groups indicating their antioxidant efficacy in protection of spermatozoa from oxidative stress. Results of study indicated that supplementation of these antioxidants to ram semen could be used to enhance storage life of liquid semen at 4°C up to 72 h. [ABSTRACT FROM AUTHOR]

Fibronectin type-II (FnII) family proteins are the major proteins in many mammalian species including bull, horse and pig. In the present study, a major FnII protein has been identified and isolated from donkey (Equus hemionus) seminal plasma, which we refer to as D onkey S eminal P lasma protein-1 (DSP-1). The amino acid sequence determined by mass spectrometry and computational modeling studies revealed that DSP-1 is homologous to other mammalian seminal plasma proteins, including bovine PDC-109 (also known as BSP-A1/A2) and equine HSP-1/2. High-resolution LC-MS analysis indicated that the protein is heterogeneously glycosylated and also contains multiple acetylations, occurring in the attached glycans. Structural and thermal stability studies on DSP-1 employing CD spectroscopy and differential scanning calorimetry showed that the protein unfolds at ~43 °C and binding to phosphorylcholine (PrC) – the head group moiety of choline phospholipids – increases its thermal stability. Intrinsic fluorescence titrations revealed that DSP-1 recognizes lyso-phosphatidylcholine with over 100-fold higher affinity than PrC. Further, interaction of DSP-1 with erythrocytes, a model cell membrane, revealed that DSP-1 binding is mediated by a specific interaction with choline phospholipids and results in membrane perturbation, suggesting that binding of this protein to sperm plasma membrane could be physiologically significant. • A new FnII family protein, DSP-1 was affinity purified from donkey seminal plasma. • DSP-1 (Mr ≈ 20 kDa) is highly homologous to other FnII proteins BSP-A1 (bovine) and HSP-1 (horse). • High-resolution LC-MS analysis indicated that DSP-1 is heterogeneously O -glycosylated. • DSC studies reveal that DSP-1 unfolds at ~43 °C and phosphorylcholine (PrC) binding stabilizes its structure. • DSP-1 perturbs erythrocyte membranes by binding to choline phospholipids and induces cell lysis. [ABSTRACT FROM AUTHOR]

Background: Kachchhi-Sindhi horses are famous for their 'Rewal chal' (a unique style of running). These horses dominantly exist in the western-northern border of India adjoining Pakistan. The breeding tract of these horses is Surat, Navsari, Kachchh district of Gujarat and Jaisalmer-Barmer districts of Rajasthan in India. Most familiar colors in the Kachchhi-Sindhi horses are bay and chestnut. Roman nose, ears curved at tips but not touching each other, short back, short pastern length, broader hoof for better grip and docile temperament are major characteristics of these horses. Methods: The present study is an effort to phenotypically characterize Kachchhi-Sindhi horse breed based on their different biometric indices for selection and conservation of stallions and mares for breeding purposes by private breeders. Fourteen biometric indices were recorded for phenotypic characterization of each breed viz., height at wither (HW), body length (BL), heart girth (HG), neck length (NL), face length (FL), face width (FW), pole, ear length (EL), ear width (EW), foreleg length (FLL), canon, pastern, hoof length (HoL) and hoof width (HoW). On average, these horses stand 148 cm height, have a body length of ~140 cm, a heart girth of ~165 cm, an ear length of ~15 cm and a face length of ~61 cm. Result: Horse keepers sustain horses in intensive as well as an extensive system of rearing. It has also been ascertained from various sources that horse number is declining rapidly, however, breed population statistics are not available. There is, therefore, an urgent need to conserve this breed. Proper managemental practices and conservation efforts will pave the way for the multiplication of this valuable equine genetic resource of India. [ABSTRACT FROM AUTHOR]

The study was conducted with the objective to evaluate the post thaw semen qualities of Poitou donkey by addition of α-tocopherol, Pentoxifylline and Taurine in semen extender. Semen was divided in four different treatments having final concentration of 150 x 106 sperm/ml with use of cryoprotectant (5% DMF) containing without supplement (control) and following supplements with concentration: á-tocopherol (1 mM), Pentoxifylline (3.5 mM) and Taurine (25 mM), all stored in liquid nitrogen for 24 hrs. All samples after thawing process were maintained at 37°C, were analyzed and performed. Spermatozoa motility, live sperm count, host reactive, acrosomal integrity and DNA integrity were recorded and performed for each additive and also for an untreated control. In between the overall treatments, Pentoxifylline (T2) and Taurine (T3) revealed the higher (P<0.01) post thaw sperm motility (53.78±0.87 and 52.28±0.90%) compared to α-tocopherol-T1 (50.15±0.80%) and control (39.65±0.52%) while, T2 and T3 differed non-significantly. Between the treatments, T2 and T3 revealed the higher (P<0.01) post thawed live sperm count (60.62±0.82and 59.46±0.84%) compared to T1 (57.12±0.77%) and control (45.43±0.56%). In between the treatments, T2 and T3 revealed the higher (P<0.01) values of post thaw host reactive sperms (43.28±0.77 and 42.18±0.84%) compared to T1 (40.21±0.72%) and control (30.15±0.40%). Whereas, T2 and T3 differed non-significantly. Between the treatments, T2 and T3 revealed the higher (P<0.01) post thaw Acrosomal integrity (87.03±0.52and 87.03±0.45%) compared to T1 (84.62±0.34%) and control (80.34±0.24%) while T2 and T3 differed non-significantly. In between the treatments (T2 and T3) revealed the higher (P<0.01) DNA integrity (88.28±0.37 and 87.50±0.28%) compared to both treatment T1 (84.75±0.26%) and control (81.03±0.37%) while T2 and T3 differed non-significantly. In above study Pentoxifylline was found to improve sperm motility, live sperm count, host reactive, acrosomal integrity and DNA integrity and may also enhance the quality of frozen equine semen by increasing the fertility rate probably. [ABSTRACT FROM AUTHOR]

Major proteins of the seminal plasma in a variety of mammals such as bovine PDC-109, equine HSP-1/2, and donkey DSP-1 contain fibronectin type-II (FnII) domains and are referred to as FnII family proteins. To further our understanding on these proteins, we carried out detailed studies on DSP-3, another FnII protein of donkey seminal plasma. High-resolution mass-spectrometric studies revealed that DSP-3 contains 106 amino acid residues and is heterogeneously glycosylated with multiple acetylations on the glycans. Interestingly, high homology was observed between DSP-1 and HSP-1 (118 identical residues) than between DSP-1 and DSP-3 (72 identical residues). Circular dichroism (CD) spectroscopic and differential scanning calorimetric (DSC) studies showed that DSP-3 unfolds at ~45 °C and binding of phosphorylcholine (PrC) – the head group moiety of choline phospholipids – increases the thermal stability. Analysis of DSC data suggested that unlike PDC-109 and DSP-1, which exist as mixtures of polydisperse oligomers, DSP-3 most likely exists as a monomer. Ligand binding studies monitoring changes in protein intrinsic fluorescence indicated that DSP-3 binds lyso-phosphatidylcholine (K a = 1.08 × 105 M−1) with ~80-fold higher affinity than PrC (K a = 1.39 × 103 M−1). Binding of DSP-3 to erythrocytes leads to membrane perturbation, suggesting that its binding to sperm plasma membrane could be physiologically significant. • Another FnII protein from donkey seminal plasma, DSP-3 was purified and characterized. • DSP-3 exhibits higher homology to equine HSP-1 than donkey FnII protein DSP-1. • High-resolution LC-MS analysis indicated that DSP-3 is heterogeneously O -glycosylated. • DSP-3 unfolds at ~45 °C and phosphorylcholine binding stabilizes its structure considerably. • DSP-3 perturbs erythrocyte membranes by specifically binding to choline phospholipids. [ABSTRACT FROM AUTHOR]

The study was aimed to superovulate donor Indian mares using eFSH. eFSH administration was initiated on 7th day of estrus cycle with dose of 25 mg @ 12.5 mg x 12 hourly or 6.25 mg x 6 hourly in total six mares. Another six mares served as control. Treatment was continued till development of two or more follicles of 35 mm size. The superovulatory response was assessed by serial ultrasonography of ovaries twice daily. Artificial insemination was undertaken in all mares with detected ovulations were detected. Similar super-ovulatory response was seen in all six mares with average days (n) of eFSH treatment of 8.6 days. Average number of follicles (>35 mm) in super-ovulated mares were 5.5. Although ovulations per mare were 4.16, yet embryo collection per mare was only 1.16 (seven embryos from six treated mares) where as in control group, it was 0.83 (five from six control mares). [ABSTRACT FROM AUTHOR]