Yuan Tian, Peng Zhang, Kemin Jing, Yuqian Li, Binglin Yue, Zhijuan Wu, Wenjing Dong, Jincheng Zhong, Xin Cai
{"title":"Overexpression of TAF4B Promoted the Proliferation of Undifferentiated Spermatogonia in Cattleyak In Vitro.","authors":"Yuan Tian, Peng Zhang, Kemin Jing, Yuqian Li, Binglin Yue, Zhijuan Wu, Wenjing Dong, Jincheng Zhong, Xin Cai","doi":"10.1111/rda.14738","DOIUrl":"10.1111/rda.14738","url":null,"abstract":"<p><p>As the hybrid between cattle and yak, cattleyak is a typical male sterile mammal, and the underlying mechanism for its spermatogenic arrest is still unclear. In this study, the coding region of cattleyak TAF4B gene was cloned by RT-PCR and analysed by bioinformatics. To investigate the effects of TAF4B on cellular proliferation and differentiation, an expression vector was generated and introduced into undifferentiated spermatogonia (UDSPG) of cattleyak. The results showed that the protein encoded by TAF4B did not contain the signal peptide sequence. The expression level of TAF4B in UDSPG of cattleyak was lower than that in yak, while the overexpression of TAF4B in cattleyak promoted the proliferation activity of cattleyak UDSPG. Meanwhile, the expression of proliferation and meiosis-related genes was increased but the differentiation-related genes were decreased. Therefore, the aberrant expression of TAF4B in cattleyak UDSPG possibly impaired its proliferation and differentiation equilibrium and decreased its growth potentiality, thereby reducing the quantity of UDSPG and affecting spermatogenesis. This study provided a potential approach for further elucidation of the mechanism of spermatogenesis arrest and provided a new idea for solving the problem of male infertility in cattleyak.</p>","PeriodicalId":21035,"journal":{"name":"Reproduction in Domestic Animals","volume":"59 11","pages":"e14738"},"PeriodicalIF":1.6,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142569366","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Carolina Tamargo, Ferran Garriga, Marc Yeste, Elisabeth Pinart, Rodrigo Muiño, María Teresa Carbajo, Carlos Olegario Hidalgo
{"title":"Predictive Indicators of Cryotolerance and Fertility in Bovine Sperm: Evaluating Fresh Semen Quality to Improve AI Outcomes With Frozen-Thawed Sperm.","authors":"Carolina Tamargo, Ferran Garriga, Marc Yeste, Elisabeth Pinart, Rodrigo Muiño, María Teresa Carbajo, Carlos Olegario Hidalgo","doi":"10.1111/rda.14742","DOIUrl":"https://doi.org/10.1111/rda.14742","url":null,"abstract":"<p><p>The success of artificial insemination (AI) with frozen-thawed semen in cattle is influenced by both female factors and sperm quality. In terms of sperm quality, prior studies indicate that the ability of frozen-thawed bovine sperm to fertilise an oocyte is dependent on their quality and resilience to cryopreservation. Cryopreservation induces oxidative stress, leading to ultrastructural damage in the sperm. This study aimed to determine whether the quality of fresh semen can identify bulls with good and poor sperm freezability. This difference between fresh and frozen semen from the same bull allows us to predict fertility. Motility and kinetic parameters were assessed using computer-assisted sperm analysis (CASA), while six functional variables were evaluated through flow cytometry, both before and after the freeze-thaw process on the sperm from 13 bulls. In vivo fertility was measured using 90-day non-return rates. The principal component analysis (PCA) of eight sperm variables post-thaw identified one principal component explaining 81.19% of the total variance and classified the bulls into two groups: Poor freezability bulls (progressive motility: 48.12% ± 8.41%; viability: 77.51% ± 7.61%) and good freezability bulls (progressive motility: 58.64% ± 6.64%; viability: 88.12% ± 2.52%). Bulls with higher freezability showed better sperm viability and motility, as well as lower levels of ROS, superoxides and intracellular calcium before cryopreservation that were significantly correlated with higher non-return rates (NRR). The results underscore the importance of assessing the quality and functionality of fresh semen to predict the fertility potential of cryopreserved sperm. This approach can aid in selecting ejaculates with the best potential for successful artificial insemination, ultimately improving reproductive performance in dairy cattle.</p>","PeriodicalId":21035,"journal":{"name":"Reproduction in Domestic Animals","volume":"59 11","pages":"e14742"},"PeriodicalIF":1.6,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142627181","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"The Effect of Different Concentrations of Melatonin on the Quality of Frozen and Thawed Semen of Jamnapari Goat.","authors":"Kabir Alam, Sushant Srivastava, Sanjeev Kumar Verma, Abhishek Kumar Verma, Saurabh, Rajesh Kumar, Rabindra Kumar, Ankit Jaiswal, Azeer Alam, Tanmoy Rana","doi":"10.1111/rda.14743","DOIUrl":"https://doi.org/10.1111/rda.14743","url":null,"abstract":"<p><p>Antioxidants help safeguard sperm cells from damage during the freeze-thaw process. Melatonin and its metabolites have an antioxidative effect. The current study aimed to evaluate the impact of melatonin supplementation in tris-based extenders at various concentrations (0.5, 1.0 and 2 mM) on the freezability of Jamnapari goat spermatozoa. A total of 48 ejaculates were collected twice a week from four Jamnapari bucks (n = 12 × 4) using an artificial vagina during the period of October to November 2023. Selected ejaculates diluted with tris-citric acid egg yolk extender were divided into four equal aliquots, and melatonin (dissolved in 0.1% dimethyl sulfoxide) was added later at 0.5, 1.0 and 2.0 mM to the control group (C) (extender and 0.1% DMSO) and treatment groups T1, T2 and T3, respectively. Various seminal attributes such as progressive motility, livability, acrosomal integrity, sperm abnormalities, sperm plasma membrane integrity, sperm penetration distance by vanguard spermatozoa in polyacrylamide gel and seminal plasma enzyme leakage (AST, ALT, ACP and AKP) were evaluated at post-dilution and post-thawed stages. Our findings revealed that all semen quality parameters were superior in melatonin-treated groups than the C, and the differences were noticeably higher in the T2 group (1.0 mM) than the other groups. Adding 1.0 mM melatonin proved to be the most effective to safeguard sperm cells from cryopreservation induced cryodamage of Jamnapari buck.</p>","PeriodicalId":21035,"journal":{"name":"Reproduction in Domestic Animals","volume":"59 11","pages":"e14743"},"PeriodicalIF":1.6,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142710913","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
M Pande, S Kumar, S Tyagi, A S Sirohi, N Chand, Y K Soni, S Mahajan, S Saha, A Sharma, Sarika, J S Rajoriya, Anjali, A K Mohanty
{"title":"Endogenous Tissue Inhibitor of Metalloproteinase-2 Levels Are Associated With High-Quality Neat Semen but Unrelated to Sperm Cryoresistance in Bulls.","authors":"M Pande, S Kumar, S Tyagi, A S Sirohi, N Chand, Y K Soni, S Mahajan, S Saha, A Sharma, Sarika, J S Rajoriya, Anjali, A K Mohanty","doi":"10.1111/rda.14741","DOIUrl":"https://doi.org/10.1111/rda.14741","url":null,"abstract":"<p><p>Tissue Inhibitor of Metalloproteinase-2 (TIMP-2) is part of the tissue inhibitors of the metalloproteinases (TIMPs) family. Its primary function is to regulate the activity of matrix metalloproteinases (MMPs) across various tissues, including those of the reproductive system. This study aimed to quantify the natural levels of TIMP-2 in seminal plasma (SP) and sperm membrane (SM) of bulls, explore potential associations between TIMP-2 levels and semen quality parameters, and examine the relationship between TIMP-2 levels and sperm cryoresistance in bulls. Thirty semen samples from Frieswal breeding bulls were categorized into two groups based on their initial progressive motility (IPM): Good (IPM ≥ 70%; n = 21) and Poor (IPM ≤ 40%; n = 9). The samples were evaluated for their quality parameters at the fresh stage, and TIMP-2 levels were measured in SP and SM using a bovine-specific ELISA kit. Following cryopreservation of Good samples (n = 21), post-thaw motility (PTM) was used to further classify samples into Freezeable (PTM ≥ 50%; n = 14) and Non-Freezable (PTM < 50%; n = 7) groups. In frozen-thawed samples, sperm attributes, kinetics, and functional parameters were assessed, and the results were correlated with retrospective TIMP-2 levels of SP/SM. Our study revealed that the quantified levels of TIMP-2 ranged from 100.27 to 535.95 ng/L in SP and from 0 to 115.78 ng/10 million spermatozoa in SM. TIMP-2 levels in both SP and SM were significantly higher in Good ejaculates compared to Poor ejaculates (p < 0.01). Furthermore, total TIMP-2 levels in the SP/SM of semen samples from bulls showed a positive correlation with fresh semen attributes. However, SP/SM TIMP-2 levels in the Freezeable group did not show any significant differences compared to the Non-Freezable group in post-thaw semen quality attributes, kinetic parameters, and functional tests, except for a significant positive correlation (r = 0.530, p < 0.05) between sperm DNA integrity and SP-TIMP-2 levels. In conclusion, the findings suggested that TIMP-2 can be a positive regulator of semen quality at the neat stage. However, when it comes to the resilience of sperm to cryopreservation, the levels of TIMP-2 do not seem to exert any significant influence in breeding bulls.</p>","PeriodicalId":21035,"journal":{"name":"Reproduction in Domestic Animals","volume":"59 11","pages":"e14741"},"PeriodicalIF":1.6,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142644686","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Matheus Cruz Silva, Mariana Moreira Dos Anjos, Higor Souza de Camargo, João Paulo Mendes Lollato, Elis Lorenzetti, Thales Ricardo Rigo Barreiros, Marcelo Marcondes Seneda, Fábio Morotti
{"title":"Use of Injectable Progesterone as a Pre-Synchronisation Strategy in a Timed Artificial Insemination Protocol Based on Gonadotropin-Releasing Hormone and Progesterone in Bos indicus Beef Cows in Anoestrous.","authors":"Matheus Cruz Silva, Mariana Moreira Dos Anjos, Higor Souza de Camargo, João Paulo Mendes Lollato, Elis Lorenzetti, Thales Ricardo Rigo Barreiros, Marcelo Marcondes Seneda, Fábio Morotti","doi":"10.1111/rda.14745","DOIUrl":"10.1111/rda.14745","url":null,"abstract":"<p><p>This study evaluated the effects of pre-synchronisation with injectable progesterone (P4) on the ovarian follicular dynamics of Bos taurus indicus cows in anoestrous treated with a timed artificial insemination (TAI) protocol. Multiparous Nelore females (n = 47) at 30-60 days postpartum were used in this study. 10 days before (D-10) the TAI protocol, antral follicle count (AFC; follicles ≥ 3 mm), ovarian condition and body condition score (BCS; 1-5) were assessed and were randomly allocated into two groups: Pre-sync (n = 25), which underwent pre-synchronisation with 150 mg of injectable P4 intramuscularly (i.m.), and control (n = 22), which received the same volume of NaCL 0.9%. On D0, the ovarian assessment was repeated, and TAI protocol was initiated in all animals, with the insertion of an intravaginal P4 device and administration of 10.5 μg of buserelin acetate (gonadotropin-releasing hormone-GnRH). On D7, the P4 device was removed, and 300 IU of equine chorionic gonadotropin, 150 μg of D-cloprostenol and 1 mg of estradiol cypionate were administered i.m. On the same day (D7), the presence of the corpus luteum (CL) was assessed, the dominant follicle was measured, and the tail was painted to evaluate estrous expression. On D9, the largest follicle was remeasured, and TAI was performed. Animals that were not detected in oestrous at the time of AI were administered 10.5 μg of GnRH i.m. Numerical data were analysed using the Mann-Whitney U test. Binary data were analysed using the Fisher's exact test (5%). BCS, both at the beginning of pre-synchronisation (p = 0.45) and TAI protocol initiation (p = 0.20), and AFC (p = 0.36) did not differ between control and Pre-sync groups. The diameter of the largest follicle was similar between the control and Pre-sync groups on D-10 (p = 0.32), D0 (p = 0.33), D7 (p = 0.29) and D9 (p = 0.22). On D7 of the protocol, the Pre-sync group had a higher percentage of CL visible on transrectal ultrasonography (84.0%; p = 0.02) than the control group (54.5%); however, the expression during oestrous did not differ between groups (p = 0.59). The pregnancy rate was similar (p = 0.64) between groups and was not influenced by the CL rate on D7 (p = 0.48), oestrous expression (p = 0.20) or their interaction (p > 0.1). Pre-synchronisation effectively increased the proportion of cows with CL on D7 without altering the diameter of the largest follicle, oestrous expression or pregnancy rate in anoestrous cows treated with a GnRH/P4-based TAI protocol.</p>","PeriodicalId":21035,"journal":{"name":"Reproduction in Domestic Animals","volume":"59 11","pages":"e14745"},"PeriodicalIF":1.6,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142644687","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Supplementing the Beltsville Extender With Mitoquinol Improves the Quality and Fertility Potential of the Rooster's Cooled Sperm.","authors":"Reihaneh Nateghi, Reza Masoudi, Nader Asadzadeh","doi":"10.1111/rda.14740","DOIUrl":"https://doi.org/10.1111/rda.14740","url":null,"abstract":"<p><p>Supplementing freeze diluents with certain antioxidants can maintain the quality of chilled sperm. The present study was an attempt to investigate the effect of Beltsville extender supplementation with the mitochondrial-targeted antioxidant 'Mitoquinol' on the quality parameters and fertility potential of rooster sperm during the cooling process. Semen samples were diluted in Beltsville extender, divided into five groups, and supplemented with 0, 1, 10, 100 and 1000 nM Mitoquinol. Samples were stored at 5°C for up to 50 h and then assayed for sperm motility, viability, mitochondrial function, membrane integrity and malondialdehyde concentration after 0, 25 and 50 h of cooling. To assess reproductive performance, artificial insemination was performed using sperm cooled for 25 h. The results showed no differences between groups at the beginning time. Extender supplementation with 10 and 100 nM Mitoquinol resulted in an improvement in total motility, progressive motility, membrane integrity, mitochondrial function and viability (p ≤ 0.05), as well as a lower malondialdehyde concentration (p ≤ 0.05) in comparison to the other groups during 25 and 50 h storage. Fertility rates were higher when roosters were inseminated with semen samples supplemented with 10 and 100 nM Mitoquinol, compared to the control group. Therefore, supplementing Beltsville extender with Mitoquinol (10 and 100 nM) effective in improving the quality and fertility potential of cooled rooster sperm.</p>","PeriodicalId":21035,"journal":{"name":"Reproduction in Domestic Animals","volume":"59 11","pages":"e14740"},"PeriodicalIF":1.6,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142558628","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mariana Garcia Kako Rodriguez, Juan Pedro Bottino, María Jesús Frisch, Marcelo Ratto, Karina Neimaur, Luis Cal-Pereyra, Aline Freitas-de-Melo, Rodolfo Ungerfeld
{"title":"Maternal-Fetal Blood Flow and Progesterone Concentration in Ewes Sheared at Mid or Late-Pregnancy.","authors":"Mariana Garcia Kako Rodriguez, Juan Pedro Bottino, María Jesús Frisch, Marcelo Ratto, Karina Neimaur, Luis Cal-Pereyra, Aline Freitas-de-Melo, Rodolfo Ungerfeld","doi":"10.1111/rda.14744","DOIUrl":"https://doi.org/10.1111/rda.14744","url":null,"abstract":"<p><p>These studies aimed to determine if shearing ewes at the second or last third of gestation modify the uterine and placentome blood flow, placentome size, and maternal progesterone concentration. Pregnant ewes were assigned to four groups of 12 ewes each according to the gestation period: mid-pregnancy sheared (on day 90 of pregnancy) or unshorn group; and late-pregnancy sheared group (on day 121 of pregnancy) or unshorn group. In both experimental periods, using spectral Doppler ultrasonography, placentomes and uterine artery blood flow and placentome size were evaluated 14 days before and 6 days after shearing. An additional measurement was performed 26 days after shearing in mid-pregnant ewes. Serum progesterone concentration was measured before shearing 4, 24, 72 h, and 22 days after shearing. The uterine artery's end-diastolic velocity (EDV) tended to be greater in the sheared than in the non-sheared ewes (p = 0.1). Peak systolic velocity (PSV) and EDV of placentome increased (p = 0.05 and p = 0.008, respectively) on day 26, accompanied by an increase in placentome area (p = 0.035) in mid-pregnant ewes. In late-pregnant ewes, uterine artery and placentome blood flow and size did not vary. Progesterone concentration varied with time but was not affected by shearing. In conclusion, shearing triggered an increase in placentome size and some changes in blood flow only when ewes were sheared during the second third of their pregnancy. Shearing ewes either the second or last third of gestation did not affect uterine artery blood flow and progesterone secretion.</p>","PeriodicalId":21035,"journal":{"name":"Reproduction in Domestic Animals","volume":"59 11","pages":"e14744"},"PeriodicalIF":1.6,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142676738","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Hussain Ahmad Saeed, Rabia Sabir, Xinyue Lu, Yuan Jiang, Bienvenu Odjoubire Mahougnon Koutonin, Dayu Wang, Yangfeng Fu, Chao Jia, Juan Li
{"title":"6-Gingerol and Astaxanthin Mitigate the Effects of Stearic Acid in Pig Oocyte Maturation.","authors":"Hussain Ahmad Saeed, Rabia Sabir, Xinyue Lu, Yuan Jiang, Bienvenu Odjoubire Mahougnon Koutonin, Dayu Wang, Yangfeng Fu, Chao Jia, Juan Li","doi":"10.1111/rda.14746","DOIUrl":"https://doi.org/10.1111/rda.14746","url":null,"abstract":"<p><p>Elevated non-esterified fatty acids (NEFAs), particularly stearic acid (SA), have a deleterious effect on oocyte maturation, leading to developmental damage and reproductive issues. High SA levels disrupt metabolic processes, inducing lipotoxicity that impairs oocyte quality and contributes to reproductive failures through early embryonic losses. This research investigates the lipotoxic effects of SA and assesses the protective potential of 6-Gingerol (6-G) and Astaxanthin (AX) on porcine oocytes during in vitro maturation (IVM). Herein, 6100 cumulus-oocyte complexes (COCs) were exposed to various concentrations of SA (25-250 μM) to elucidate the concentration-dependent effect on oocyte viability, polar body extrusion (PBE) and cumulus cell expansion index (CCEI). However, the efficacy of 6-G (5-15 μM) and AX (2.5 μM) in combination with SA at 150 μM (SA6) concentration was evaluated to mitigate these adverse effects. The results indicated that SA6 substantially reduced oocyte viability, PBE and CCEI, demonstrating its toxic impact on oocyte developmental competence (p < 0.0001). Moreover, treatment with antioxidants such as SA6 + 6-G (10 μM) and SA6 + AX showed a considerable increase in viability and PBE compared to SA6 alone (p < 0.05). These findings demonstrate the importance of lipid metabolism in oocyte health, where dysregulation impairs reproductive capacity. Both 6-G and AX protected against lipotoxicity induced by SA6 while enhancing lipid homeostasis and the anti-oxidative defences necessary for maintaining cellular integrity. This study finds substantial evidence that optimising the microenvironment with specific antioxidants can improve oocyte quality and provide invaluable knowledge in reproductive technologies and fertility treatments.</p>","PeriodicalId":21035,"journal":{"name":"Reproduction in Domestic Animals","volume":"59 11","pages":"e14746"},"PeriodicalIF":1.6,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142710829","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Meeti Punetha, Sheetal Saini, Surabhi Sharma, Swati Thakur, Priya Dahiya, Manu Mangal, Rajesh Kumar, Dharmendra Kumar, P S Yadav
{"title":"CRISPR-Mediated SRY Gene Mutation Increases the Expression of Female Lineage-Specific Gene in Pre-Implantation Buffalo Embryo.","authors":"Meeti Punetha, Sheetal Saini, Surabhi Sharma, Swati Thakur, Priya Dahiya, Manu Mangal, Rajesh Kumar, Dharmendra Kumar, P S Yadav","doi":"10.1111/rda.14739","DOIUrl":"https://doi.org/10.1111/rda.14739","url":null,"abstract":"<p><p>In mammals, sex determination is governed by the SRY gene on the Y chromosome, redirecting gonadal development from forming ovaries to testes. Mutations or alterations in the SRY gene can significantly affect phenotypic changes and lineage-specific markers. This study aims to elucidate the role of the SRY gene in buffalo embryos using CRISPR-Cas9 technology. We designed a crRNA targeting the HMG domain of the SRY gene using the CRISPOR algorithm. Nucleofection of sgRNA-Cas9 RNPs into buffalo fibroblasts confirmed efficient cleavage at the targeted site. Using this validated guide, we investigated the role of the SRY gene in sexual determination by electroporating CRISPR-Cas9-RNPs into single-stage zygotes of buffalo. Genetic changes in the SRY gene were confirmed through sequencing, revealing mosaic blastocysts with multiple alleles and non-mosaic mutants. Mutations in SRY gene increased the expression of female lineage-specific gene Wnt4 whereas decreased the expression of male specific gene SOX9 in blastocysts, suggesting reprogramming towards female sex determination pathways. Our findings provide insights into buffalo sex differentiation mechanisms and potential applications in reproductive strategies for breeding programmes.</p>","PeriodicalId":21035,"journal":{"name":"Reproduction in Domestic Animals","volume":"59 11","pages":"e14739"},"PeriodicalIF":1.6,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142584155","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Juliana Pedrosa, Camila Ezepha, Felipe Anibal Carvalho-Costa, Maria Isabel Nogueira Di Azevedo, Walter Lilenbaum
{"title":"A Neglected Part of Bovine Genital Leptospirosis: Bulls.","authors":"Juliana Pedrosa, Camila Ezepha, Felipe Anibal Carvalho-Costa, Maria Isabel Nogueira Di Azevedo, Walter Lilenbaum","doi":"10.1111/rda.14747","DOIUrl":"https://doi.org/10.1111/rda.14747","url":null,"abstract":"<p><p>Bovine leptospirosis is a worldwide disease that causes reproductive issues, including early embryonic death, stillbirths and infertility, which result in significant economic losses. Although bovine leptospirosis is well-documented in cows, the role of bulls in harbouring and potentially transmitting genital leptospirosis has been largely neglected. We examined genital and blood samples from 16 slaughtered bulls using microscopic agglutination testing (MAT), polymerase chain reaction (PCR) and sequencing of amplicons. Our results showed that 81.2% of bulls were seroreactive, and 75% were genitally infected. The amplicons displayed an identity greater than 99% with Leptospira interrogans strains from the Sejroe serogroup, specifically serovar Hardjoprajitno. This study demonstrates that bulls can harbour in their genital tract the same leptospires associated with reproductive syndromes in cows from the same geographic region, highlighting the importance of bulls in maintaining and transmitting Sejroe serogroup strains associated with reproductive disease.</p>","PeriodicalId":21035,"journal":{"name":"Reproduction in Domestic Animals","volume":"59 11","pages":"e14747"},"PeriodicalIF":1.6,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142710911","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}