<div style="text-align: justify;">Taking into account that punicalagin (PUN) is a very powerful antioxidant; the current study evaluated the potential positive effects of oral PUN on spermatologic parameters of male New Zealand White rabbits. A total of 24 male bucks was housed individually and trained for semen collection. After the training for 2 wk, rabbits were assigned into 4 groups and received daily gavages of 0, 1, 2, and 10 mg/kg PUN in tap water for 9 wk. Semen was collected weekly from each rabbit and samples at d 1 and 63 of the experiment were analyzed separately. Initial values (ejaculate volume, ejaculate weight, ejaculate pH, sperm concentration, percent progressive motility, and seminal plasma protein levels) tested at d 1 were similar among the groups. &nbsp;There were also no differences in ejaculate volume, ejaculate weight, ejaculate pH and seminal plasma protein concentrations at the end of the experiment. Libido and reproductive organ weights were not affected by the treatments.However, sperm concentrations (P&lt;0.01) and percent progressive motility (P&lt;0.04) were significantly improved for bucks in 2 and 10 mg/kg PUN groups. Thus, the current study suggested that as low as 2 mg/kg/day PUN can be beneficial for spermatogenesis and motility in male rabbits.</div>

Punicalagin is beneficial for spermatogenesis in male New Zealand White rabbits

<div style="text-align:justify">Background: Chinese pond turtle (CPT, Mauremys reevesii) is a typical species with temperature-dependent sex determination (TSD). Pond slider turtle (PST, Trachemys scripta) is considered one of the worst invasive species in the world. In order to analyze the adaptive evolution of fecundity traits in CPT and TSD, the rapidly evolving gene families were obtained and functional enrichment analysis was performed. At the same time, CPT and PST species-specific genes were also enriched for their fecundity traits. 
 
Methods: Orthofinder software was employed to search homologous genes based on protein sequence. CAF&Eacute; software was used to obtain the expansion and contraction gene families of CPT and PST. Then, GO terms and pathway enrichment analyses were performed using TBtools software and Swissprot database.&nbsp; 
 
Result: In CPT, 159 rapid expansion gene families GO enrichment results showed that the reproductive activities related pathways were mainly three significant enrichment process, spermatogenesis, steroid hormone mediated signaling pathway and male gamete generation. 2 rapidly contraction gene families showed that there was no significant enrichment pathway related to reproduction. The pathway of significant enrichment were RNA biosynthetic process, regulation of RNA metabolic process and regulation of cellular metabolic process. The gene families unique to the species were significantly enriched in pathways DNA replication-dependent nucleosome organization, DNA replication-dependent nucleosome assembly and negative regulation of megakaryocyte differentiation. In PST, 15 rapid expansion gene families GO enrichment results showed that the most significant GO terms mainly consisted of biological processes which were involved in endocrine process, regulation of endocrine process and cellular response to steroid hormone stimulus, but spermatogenesis and male gamete generation gene families also expanded. 58 rapidly contraction gene families showed that the pathways of significant enrichment were endocrine process, cellular response to steroid hormone stimulus and regulation of endocrine process. At the same time, estrogen secretion, binding of sperm to zona pellucida and sperm-egg recognition gene family also rapidly contraction. The gene families unique to the species were significantly enriched in pathways adenylate cyclase-activating G protein-coupled receptor signaling pathway, double-strand break repair and regulation of potassium ion transport.</div>

Comparison of Fecundity Traits between Mauremys reevesii and Trachemys scripta based on Gene Family Analysis

Behavioral, hormonal and histological changes caused by heat stress at 30 and 40oC were studied in mice model. As a result, obnoxious activities like restlessness and hyperactivity were observed. Rectal temperature was significantly increased at 40oC as well as the increase in water intake, whereas, the feed intake remained consistent. Body weight significantly decreased at 40oC. Serum testosterone has shown a decrease in both groups. At day 7 of the treatment, low levels of growth hormone were recorded whereas no significant difference was recorded at day 15. Adrenocorticotropic hormone level was significantly decreased in both groups at day 7. Shrinking of the seminiferous tubules with irregular appearance containing less spermatozoa was also observed at 40oC. Testicular degeneration and atrophy of leydig cells were also observed. Finally, we concluded that heat stress adversely affects the male reproductive functions along with hormonal imbalances and decline in the spermatogenesis.&nbsp;

Thermally induced stress revealed behavioral, hormonal&nbsp;and histological disturbances in mice model&nbsp;

<div>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.</div>

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

<div>Background: Ets variant factor 5 (ETV5) plays an important regulatory role in mouse Spermatogonial stem cells (SSCs) self-renewal. ETV5 knockout (KO) mice exhibit a progressive loss of SSCs and resulting in a Sertoli cell-only phenotype. The current study was aimed to use gene editing technology to obtain ETV5-KO pigs as a model for studying the apoptosis mechanism of SSCs and further clarify the function of ETV5 gene in pigs. 
 
Methods: A gene editing plasmid for the porcine ETV5 gene was constructed, transfected into porcine fetal fibroblasts by electroporation to obtain ETV5-KO cells. ETV5-KO cells were used as donors to prepare ETV5-KO pigs by somatic cell nuclear transfer (SCNT). Testis tissues were collected for hematoxylin and eosin (HE), immunohistochemistry (IHC), RT-PCR testing and blood for ELISA testing from ETV5-KO pig. 
 
Result: In the present study, we used the CRISPR/Cas9 system and SCNT to generate homozygous ETV5-KO pigs. We observed 3 phenotypes in these pigs: normal testis development after birth, the SSCs in the seminiferous tubules did not show obviously extinction at sexual maturity and normal spermatogenesis.</div>

<div>Generation of ETV5 Knockout Pigs with CRISPR/Cas9</div>

<div style="text-align: justify;">Bull fertility is an important factor for genetic improvement; therefore prediction of fertility an early age of the bulls by using genetic markers is a goal for livestock breeding. Zinc finger protein280BY linked gene is predominantly expressed in testis and has a major role in spermatogenesis. An investigation was carried out to detect the genetic variants in the male specific region (MSY) of ZNF280BY gene and their association with semen quality traits in Murrah buffaloes. Polymerase Chain Reaction-Single Strand Conformation Polymorphism technique (PCR-SSCP) and DNA sequencing methods revealed three different SSCP band patterns (AA, BB, CC); one transition (C10375T) &nbsp;and an indel (insertion of T at 10460-10461). The associations of the genetic variants with semen volume, sperm concentration, Hypo Osmotic Swelling Test (HOST) in fresh and frozen semen were observed. AA and CC genotype bulls had high volume (P&pound;0.05) of semen per ejaculation than BB genotype bulls. BB genotype bulls had high sperm concentration (106/mL) per ejaculate in comparison to AA and CC genotype bulls (P&pound;0.05). AA and BB genotype bulls had a higher per cent of HOST (P&pound;0.05) reacted sperms of fresh semen and per cent acrosomal integrity of frozen semen in comparison to CC genotype bulls. The observed association between SSCP variants in MSY region of ZNF280BY gene with semen quality parameters indicated the possibilities of using ZNF280BY as a candidate gene for identification of markers for semen quality traits in Murrah buffaloes.</div>

<div style="text-align: justify;">Genetic variants in male specific region (MSY) of ZNF280BY gene and their association with semen quality traits in Murrah buffalo bulls</div>

<div style="text-align: justify;">Bull fertility is an important component of reproductive efficiency in dairy cattle. PPP1R11 is a potent candidate gene for bull fertility being involved in the pathway of spermatogenesis. Present investigation was carried out in twenty Karan Fries (KF) bull to identify SNPs of PPP1R11 gene and their association with conception rate. Genomic DNA was extracted from frozen semen straws by phenol-chloroform method. Primer pair targeted to exonic II region of PPP1R11 gene was designed using Primer 3.1 software. PCR reactions were carried out in thermal cycler after optimizing. The PCR product was checked on 1.5% horizontal agarose gel electrophoresis to verify the amplification of target region. Samples were sequenced and SNPs were identified using multiple alignment programme with respect to reference sequence and visual inspection of chromatograms. Sequencing results revealed only two SNPs [G28708846A, T28708860C] at exonic II region of PPP1R11 gene in KF bulls. Each bull was genotyped with respect to each of the two identified SNPs. Association of SNP genotype with conception rate was analyzed using least square ANOVA taking genotypes as fixed effect. The result indicated that, only one SNP [T28708860C] had significant (p&lt;0.01) effect on conception rate. The bulls with TT genotype revealed significantly (p&lt;0.01) highest conception rate (53.20&plusmn;2.96%) followed by bulls with TC (43.12&plusmn;2.39%) and the lowest in bulls with CC (34.59&plusmn; 2.28%) genotypes. The results of present finding suggested that association of SNPs with conception rate after validation on large population might be useful in marker assisted selection for selection of bull with high fertility in future.</div>

Association of single nucleotide polymorphisms of PPP1R11 gene with conception rate in Karan Fries bull

Background: The orexin (OX) is a hypothalamic neuropeptide that exists in two forms, orexin A (OXA) and orexin B (OXB), which bind two receptors (OX1 and OX2). While orexin B shows selectivity for OX2, OXA is potent at both receptors. Orexins are also reported in other mammalian organs where they undergo many regulatory functions. However, their detection in the dromedary camel reproductive organs remains to be revealed. 
 
Methods: We investigated the immunoreactivity of orexin OXA and OX1 in the testis of young (6-12 month), pre-pubertal (2-3 years) and adult (6-15 years) dromedary camels. We also studied the relative expression of hypocretin neuropeptide precursor (HCRT) in the testicular tissue of these age groups. 
 
Result: Positive immunoreactions of OXA and OX1 were shown in both the tubules and interstitium of dromedary camel testis in the young animals. While positive OXA and OX1 immune-labeling was detected in the spermatogonia, Sertoli cells and Leydig cells in all ages, adult camels also showed positive OXA and OX1 signals in the primary spermatocytes, spermatozoa and interstitial blood vessels. In testes at pre-pubertal and adult stages, spermatogenic cells, Sertoli cells and Leydig cells showed stronger OXA and OX1 reactions than in young camels. Real time PCR analysis also revealed that HCRT was expressed in the testis of dromedary camels with minimal expression in young camels. In conclusion, the current study provides the first evidence for the presence of OXA and OX1 in the testis of adult and younger dromedary camels. Before puberty, the binding of OXA and OX1 in the camel testis might be important for development and proliferation of spermatogenic cells, Sertoli cells and Leydig cells; in adult animals they may play a role in steroidogenesis and spermatogenesis.

Age-related Localization and Expression of Orexin A and its Receptor 1 in Dromedary Camel Testis&nbsp;

<div>Background: Pesticides act as endocrine disruptors and cause enormous disturbances in steroidogenesis, spermatogenesis and sexual behavioural display. These toxicants modulate and/or disrupt the reproductive and hormonal environment by acting on hypothalamus, pituitary and reproductive organs. Few studies have been carried out to assess the levels of pesticides in dairy cattle / buffaloes. The information on pesticide residues in body fluids especially blood and semen in breeding bulls is lacking.&nbsp; 
 
Method: In the light of above background nineteen fresh semen ejaculates were collected from Milkfed, Khanna and PLDB (Punjab Livestock Development Board), Nabha, Punjab and stored at -80◦C. Immediately after collection, the semen was centrifuged at 4500 &times; g for 20 min at 4◦C. The supernatant (seminal plasma) was stored at -20◦C until processing for the assessment of pesticide residues using gas chromatography (GC). In the present study on crossbred breeding bulls, the semen samples were analyzed for organochlorines, organophosphates and synthetic pyrethroid pesticide residues. Seven OCP were screened namely Heptachlor epoxide, Chlordane, Fipronil, Lindane, Methoxychlor, op-DDT, Endrin. Eleven OPP were screened namely Chorpyrifos, Dichlorovos, Ethion, Monocrotophos, Malathion, Parathion-methyl, Profenphos, Phorate, Triazophos, Quinalphos and Phosalone and four SP&rsquo;s namely Cypermethrin, Permethrin, Deltamethrin, Cyalothrin were screened in the semen samples. Semen samples were also assessed for viability, HOST (%) and abnormality (%) for both fresh as well as post thaw samples. 
 
Result: None of the semen samples were found positive for any of the above pesticide residues. This may be attributed to the feeding of fodder cultivated at the stations without using any pesticides, hence no pesticide residue found in the semen samples of the crossbred breeding bulls of these semen stations In present study, viability (%) of sperms were found to be significantly higher (p&lt;0.05) in fresh (75.57 &plusmn; 5.31) as compared to post thaw (70.17 &plusmn; 4.64) semen of bulls. The abnormality (%) of sperms was found to be significantly lower (p&lt;0.05) in fresh (13.61 &plusmn; 3.71) than post thaw (18.22 &plusmn; 3.59) semen bulls. Also in present study the host (%) was found 67.23&plusmn;4.76 in fresh semen which decreased to 46.82&plusmn;3.55 in post thaw semen.</div>

​Evaluation of Level of Pesticide Residue in Seminal Fluid of Crossbred Bulls

<div>Background: Bio potency of trace elements is very important in animal growth and reproduction. Among the trace minerals, zinc is very important as it is required for many normal physiological functions. It also plays an essential role in testicular steroidogenesis androgen metabolism and interaction with steroid receptors. Testes have a strict zinc requirement and severe zinc deficiency compromises spermatogenesis, sperm viability, motility and fertility in human beings. Zinc depletion studies in mice resulted in oligospermia with poor Leydig cell function and lowered testosterone concentration and was reversed by zinc supplementation. But, relatively few studies have been conducted on male animal responses to supplementation of important mineral like zinc at their critical phase of growth and reproduction. The present investigation was taken up to study the effect of zinc supplementation in two different doses with a view to assess its implication in testicular and epididymal sperm morphology during growth period in Wistar rats. 
 
Methods: The study was conducted in 72 weaned Wistar male rats for a period of 8 weeks from 4 to 12 weeks of age, which were procured from Indian Institute of Integrative Medicine, CSIR Laboratory, Jammu. The experimental rats were divided in to three groups as control: rats fed diet without zinc supplementation, T1 (treatment 1): rats fed diet containing zinc sulphate @ 50 mg/kg body weight/day and T2 (treatment 2): rats fed diet containing zinc sulphate @ 100 mg/kg body weight/day. They were provided standard pelleted ration and clean drinking water ad libitum and maintained under standard managemental conditions. On 6, 8, 10 and 12 weeks of experiment, the rats were sacrificed after induction of proper anaesthesia. The reproductive organs (testes and epididymidis) were collected from each rat and tissue pieces from testes and three parts of the epididymis viz. caput, corpus and cauda, were fixed separately in 10 per cent Neutral Buffered Formalin solution and processed for paraffin sections by alcohol- xylene method. Sections 5 &mu; thickness was stained by Haematoxylin and Eosin (Luna, 1968) for histological studies. 
 
Result: Elongated spermatids were seen in the seminiferous epithelium only in treatment (T1 and T2) groups at 6 weeks of age, not in the control rats. At 8 weeks of age, maximum number of spermatozoa were seen in the seminiferous tubular lumen in the T1 group as compared to T2 treatment group. At 10 weeks, spermatozoa were seen in the lumen of the seminiferous tubules in all the rats under study. The epididymis revealed no sperm pack in cauda epididymis in all group rats at 6 weeks of age. However, at 8 weeks of age, the sperm pack was seen in the lumen of the epididymal tubules of the cauda epididymis of T1 and T2 groups. The significant finding was that in T1 rats the sperm pack was very compact at 10 weeks in cauda epididymis as compared to T2 and control group rats.</div>

Punicalagin is beneficial for spermatogenesis in male New Zealand White rabbits

Comparison of Fecundity Traits between Mauremys reevesii and Trachemys scripta based on Gene Family Analysis

Thermally induced stress revealed behavioral, hormonal and histological disturbances in mice model

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

Generation of ETV5 Knockout Pigs with CRISPR/Cas9

Genetic variants in male specific region (MSY) of ZNF280BY gene and their association with semen quality traits in Murrah buffalo bulls

Association of single nucleotide polymorphisms of PPP1R11 gene with conception rate in Karan Fries bull

Age-related Localization and Expression of Orexin A and its Receptor 1 in Dromedary Camel Testis

Evaluation of Level of Pesticide Residue in Seminal Fluid of Crossbred Bulls

Effect of Zinc Supplementation on Histomorphology of Testis and Epididymis in Male Wistar Rats