Reproduction in Domestic Animals最新文献

Ultrasonic vibration (USV) of semen extender can be a novel method to modify the physico-chemical properties of semen extenders and improve their cryoprotective quality. This property may be further improved by combining with antioxidants. Therefore, this study examined the impact of a USV-treated extender enriched with Myo-inositol (MYO) on frozen ram sperm quality in a completely randomised block design with a 2 × 2 factorial arrangement. The main factors were USV (without and with) and MYO (0 and 7.5 mM) arranged in four treatments. Ultrasonic groups' diluents vibrated at 50% power for 15 min. In the breeding season, semen was collected from four Mehraban rams once a week, over five consecutive weeks. Ejaculates were mixed and divided equally into four parts and diluted up to the concentration of 200 × 10⁶ spermatozoa/ml. The samples were frozen in liquid nitrogen. At least 72 h after freezing, samples were thawed and subjected to evaluation. Eosin stain, hypo-osmotic swelling test, computer-assisted sperm analysis, ferric reducing antioxidant power (FRAP), and thiobarbituric acid assay were methods for evaluation of viability, functional integrity of plasma membrane, sperm motility, total antioxidant capacity and lipid peroxidation respectively. Intracellular ROS level and mitochondrial membrane integrity were detected using 2'-7'-dichlorodihydrofluorescein diacetate (DCFH-DA) and Rhodamine 123 coupled with propidium iodide (PI) stain by flow cytometry. The interaction effects of USV × MYO for none of the studied parameters were significant. Ultrasonic vibration increased viability, total and progressive motility and several kinematic factors; however, sperm antioxidant capacity was reduced and intracellular H₂O₂ level increased slightly (p < 0.05) without impaired lipid peroxidation or mitochondrial membrane integrity. Total and progressive motility was improved with MYO (p < 0.05). The values of VCL and VAP tended (p < 0.1) to increase with MYO treatment. The highest values of total motile and progressively motile spermatozoa and VCL were observed in the USV group enriched with MYO (p < 0.05). In conclusion, optimal ram sperm function can be preserved in the USV-treated or MYO-supplemented extenders. However, MYO enrichment of the USV-treated extender has no additional effect.

The following case report describes two pregnancies contralateral to the side of the corpus luteum (CL) that were carried to term after artificial insemination (AI) in Holstein heifers. Pregnancy diagnosis was performed by ultrasonography at 32, 46 and 74 days after timed AI. Both heifers had the pregnancy located within the right uterine horn and only one CL on the left ovary. Another eight lactating cows with contralateral pregnancies were followed around the same time; however, all lost the pregnancies between 32 days after AI and 74 days after AI. Pregnancies contralateral to the side of the CL have been described after embryo transfer in heifers, but to our knowledge, this is the first report of contralateral pregnancies after AI that were carried to term in heifers. In previous reports, all seven cows with contralateral pregnancies lost the pregnancies by 49 days of gestation. It is not clear why contralateral pregnancies can be successful in heifers but not in lactating Holstein cows. In summary, we confirmed that contralateral pregnancies were not viable in lactating Holstein cows but that successful contralateral pregnancies after AI are possible in Holstein heifers. The mechanism for this finding deserves further investigation.

This study assessed the potential protective effects of Mangiferin (MANGI) at various concentrations (10, 50 and 100 μM) on follicular survival and activation, enzyme activity of catalase (CAT) and superoxide dismutase (SOD), as well as the relative mRNA expression for BAX, BCL2 and SOD in porcine ovarian tissue cultured in vitro for 48 h with and without chemical stress induced by 0.3 μg/mL doxorubicin (DOX). The results indicated that when MANGI was associated with DOX, regardless of the tested concentration, MANGI increased (p < 0.05) follicle survival when compared to DOX alone, with only MANGI 10 + DOX and MANGI 50 + DOX treatments reducing follicular degeneration to levels comparable with the culture control. Regarding antioxidant enzymes, all treated groups decreased (p < 0.05) SOD activity compared to cultured control, while MANGI 10 treatment showed lower (p < 0.05) CAT activity compared to DOX treatment. Compared to cultured control, the mRNA levels of SOD were reduced (p < 0.05) in the MANGI 100, MANGI 10 + DOX and MANGI 100 + DOX treatments. In conclusion, supplementing the culture medium with MANGI attenuated the toxicity induced by DOX during the in vitro culture of pig preantral follicles enclosed in ovarian tissue.

Testicular vitrification requires the use of high concentrations of cryoprotectants, which can cause damage to samples due to their toxicity. The combination of these substances comes up as a way to mitigate this problem. Thus, the aim of this study was to evaluate three cryoprotectant combinations in the testicular vitrification of dogs. Ten testicular pairs from adult dogs were used, from which 12 fragments of each pair were obtained, distributed among the fresh control group (CTR) and the experimental groups according to the cryoprotectant combinations tested: dimethyl sulfoxide (DMSO)/ethylene glycol (EG), DMSO/glycerol (GLY), and EG/GLY. The fragments were vitrified using the solid surface vitrification method (SSV), at a final concentration of 5.6 mol/L of the combined cryoprotectants. Subsequently, they were warmed up and processed for histomorphological morphometric evaluations and determination of mitochondrial activity with Rhodamine 123. Considering the morphological evaluation, the DMSO/EG group showed results similar to CTR, with good scores for nuclear integrity and cell organisation in the seminiferous tubules (p > 0.05). In contrast, the EG/GLY group presented greater nuclear condensation. It was difficult to visualise and distinguish between spermatogonia and Sertoli cells (p < 0.05). The DMSO/GLY group also showed distinct levels between spermatogonia and Sertoli cells, as well as nuclear condensation, which statistically differed from CTR (p < 0.05). Also, it was observed a random distribution of the remaining cells in the seminiferous tubules of the EG/GLY and DMSO/GLY groups. The three tested groups showed basement membrane retraction and a reduction of approximately 11.6% in the average diameter of the seminiferous tubules (p < 0.05). Vitrification did not influence the mitochondrial activity of the samples, regardless of the combination of cryoprotectants used (p > 0.05). It was concluded that the DMSO/EG combination best contributed to the maintenance of the testicular histomorphological structure of dogs after vitrification.

This study evaluated the effects of subclinical pregnancy toxaemia (SPT) on fetal skeletal muscle development and assessed the potential protective role of L-carnitine supplementation during gestation. A total of 18 crossbred Hamdani ewes underwent oestrous synchronisation, natural mating and pregnancy confirmation via ultrasonography on day 45 post-mating. The ewes were managed according to NRC (2007) dietary guidelines until day 100 of gestation, after which they were assigned to three experimental groups: subclinical PT group (group 1; G1, n = 6), treatment group (subclinical PT + L-carnitine, group 2; G2, n = 6) and control group (group 3; G3, n = 6). Blood β-hydroxybutyrate (βHBA) concentrations were measured on day 100 and 138 of the gestation. Then, all ewes were slaughtered for fetal muscle sampling from the Musculus Longissimus Dorsi (MLD) and Vastus Lateralis (VL). Results indicated a significant reduction in muscle fibre number and fibre diameter in both MLD and VL in the SPT group (G1) compared to the control (G3) (p < 0.05). No significant differences were observed between G1 and G2 or between G2 and G3 for these parameters (p > 0.05). On the other hand, large effect sizes for group and pairwise comparisons imply that SPT may negatively affect prenatal muscle development and L-carnitine supports muscle development during the prepartum period. These findings highlight the negative effects of SPT and protective effects of L-carnitine supplementation on fetal skeletal muscle development in ewes with SPT. The observed deficits may negatively impact postnatal growth, survival rates and meat quality. Further investigations are warranted to optimise maternal nutrition strategies and evaluate therapeutic interventions aimed at mitigating the adverse impacts of SPT on fetal muscle development in ruminants. Furthermore, L-carnitine supplementation may be a useful in compensating for the negative effects of SPT.

Equine Coital Exanthema (ECE) is an endemic herpesvirus disease primarily affecting the external genitalia and impairing mating activities in horses. Its extremely contagious nature, latency and subclinical features can result in outbreaks and significant economic losses. Transmission occurs primarily through mating activities; therefore, robust biosecurity measures are crucial in breeding facilities. This study aims to determine the clinical prevalence of ECE among horses in a covering station in Türkiye from 2021 to 2024. It also aims to assess the efficacy of routine PCR implementation within ECE's control strategies. A cross-sectional study design has been employed. Genital swab samples were collected from clinically suspected horses, which were tested for EHV-3 using real-time PCR. Animal records, clinical examination data and PCR test results were obtained from horses at the covering station between 2021 and 2024. During the 4 years (2021-2024), 9231 mating activities were carried out, and a total of 228 clinically suspected horses were tested for EHV-3 using real-time PCR. Among these 228 horses, 6 horses (2.6%) were confirmed positive for EHV-3. The primary weakness of this study is the failure to detect subclinical cases with PCR. The absence of follow-up PCR testing in two clinically infected horses represents a limitation of this study. The molecular diagnosis of ECE was reported for the first time in Türkiye. Clinical ECE cases infrequently transpired over the four-year period at the covering station. No outbreak transpired during this interval. PCR testing plays a crucial role in disease control when implemented with suitable management methods. Additional global epidemiological investigations on ECE are required.

Pre-implantation embryo biopsy is a well-established technique in large livestock like cattle. However, its application in porcine embryos remains relatively limited. In this study, we successfully obtained trophectoderm cells from porcine IVP blastocysts using laser-assisted microdissection. First, we obtained trophectoderm cells of varying cell numbers (Group 1: less than three cells; Group 2: more than five cells) using laser-assisted microdissection and then evaluated the impact of this dissection procedure on embryo morphology. The morphological observation confirmed the complete normal post-biopsy recovery of blastocysts in the two groups. Furthermore, the expression of proteins associated with cell junction and structural integrity remained unaffected. Comparative analysis of blastocyst cell numbers and apoptotic rates among Group 1, Group 2 and the control group further validated the minimalinvasiveness of the laser-assisted microdissection when cutting less than three cells. The quality and utility of the biopsied trophectoderm cells were confirmed through successful sex identification and transcriptome sequencing. In conclusion, this study successfully established a minimal invasive laser-assisted microdissection technique for porcine blastocysts, enabling pre-implantation sex identification and blastocyst quality assessment.