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Contemporary systems for oocyte retrieval and culture of both cattle and human embryos are suboptimal with respect to pregnancy outcomes following transfer. In humans, chromosome abnormalities are the leading cause of early pregnancy loss in assisted reproduction. Consequently, pre-implantation genetic testing for aneuploidy (PGT-A) is widespread and there is considerable interest in its application to identify suitable cattle IVP embryos for transfer. Here we report on the nature and extent of chromosomal abnormalities following transvaginal follicular aspiration (OPU) and IVP in cattle. Nine sexually mature Holstein heifers underwent nine sequential cycles of OPU-IVP (six non-stimulated and three stimulated cycles), generating 459 blastocysts from 783 oocytes. We adopted a SNP-array approach normally employed in genomic evaluations but reanalysed (Turner et al., 2019; Theriogenology125: 249) to detect levels of meiotic aneuploidy. Specifically, we asked whether ovarian stimulation increased the level of aneuploidy in either trophectoderm (TE) or inner-cell mass (ICM) lineages of blastocysts generated from OPU-IVP cycles. The proportion of Day 8 blastocysts of inseminated was greater (P, Highlights • In vitro oocyte maturation rather than ovarian stimulation contributes to aneuploidy. • Trophectoderm biopsies accurately represent the ploidy status of the overall embryo. • SNP-array analyses facilitate simultaneous genomic evaluation and aneuploidy screening. • Incidence of aneuploidy declines in developmentally more advanced embryos. • High degree of variability in the incidence of aneuploidy exists between donors.

Article Details: Received: 2020-09-29 | Accepted: 2020-11-27 | Available online: 2021-01-31 https://doi.org/10.15414/afz.2021.24.mi-prap.15-19 Over the past decades natural substances are widely used in the maintaining of spermatozoa viability. The target of present study was to evaluate the effect of various taurine concentrations on stallion spermatozoa during 37°C cultivation. Fresh semen was collected from 10 breeding stallions. The experimental groups were supplemented with six different concentration of taurine (in mg/ml): A – 2.5, B – 5, C – 7.5, D – 10, E – 15, F – 20 and compared to control (CON – 0). Spermatozoa motility was assessed using the Computer Assisted Semen Analyzer (CASA) system in 6 time periods (0, 1, 2, 3, 4 and 5 hours). The MTT test was used for detection of viability. For measuring antioxidant activity FRAP and TOS methods were used. Significantly negative effect was observed in the samples with the highest concentration of taurine (20 mg/ml). Spermatozoa viability was not significantly affected in analysed concentrations of taurine. Significant higher antioxidant activity was detected in the sample with the highest taurine concentration. Data clearly showed negative effects of high taurine concentrations on stallion spermatozoa. Keywords: taurine, CASA, antioxidant activity, spermatozoa, stallion References Benzie, I. F. and Strain, J. J. (1996). The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: The FRAP assay. Analytical Biochemistry , 239 (1), 70-76. https://doi.org/10.1006/abio.1996.0292 Bucak, M. N. et al. (2007). The influence of trehalose, taurine, cysteamine and hyaluronan on ram semen: Microscopic and oxidative stress parameters after freeze–thawing process. Theriogenology , 67 (5), 1060-1067. https://doi.org/10.1016/j.theriogenology.2006.12.004 Erel, O. (2005). A new automated colorimetric method for measuring total oxidant status. Clinical Biochemistry , 38 (12), 1103-1111. https://doi.org/10.1016/j.clinbiochem.2005.08.008 Halo, M. and Tirpak, F. (2018) Stallion fertility - the basis of successful reproduction. Svet koni . 18. Halo Jr., M. et al. (2019) Time and dose-dependent effects of Viscum album quercus on rabbit spermatozoa motility and viability in vitro. Physiological Research, 68 (6), 955-972. https://doi.org/10.33549/physiolres.934223 Ijaz, A. and Ducharme, R. (1995). Effect of various extenders and taurine on survival of stallion sperm cooled to 5 C. Theriogenology , 44 (7), 1039-1050. https://doi.org/10.1016/0093-691x(95)00290-o Jambor, T. et al. (2017) In vitro effect of 4-nonylphenol on human chorionic gonadotropin (hCG) stimulated hormone secretion, cell viability and reactive oxygen species generation in mice Leydig cells. Environmental Pollution , 222, 219–225. https://doi.org/10.1016/j.envpol.2016.12.053 O'flaherty, L. et al. (1997) Intestinal taurine transport: a review. European Journal of Clinical Investigation , 27 (11), 873-880. https://doi.org/10.1046/j.1365-2362.1997.2000747.x Reddy, N. S. S. et al. (2010). Effects of adding taurine and trehalose to a tris-based egg yolk extender on buffalo (Bubalus bubalis) sperm quality following cryopreservation. Animal Reproduction Science , 119 (3-4), 183-190. https://doi.org/10.1016/j.anireprosci.2010.01.012 Slanina, T. et al. (2018) Effect of taurine on turkey (Meleagris gallopavo) spermatozoa viability and motility. Czech Journal of Animal Science , 63 (4),127-135. https://doi.org/10.17221/79/2017-CJAS Stephens, T. D. et al. (2013) Effects of pentoxifylline, caffeine, and taurine on post-thaw motility and longevity of equine frozen semen. Journal of Equine Veterinary Science , 33 (8), 615-621. https://doi.org/10.1016/j.jevs.2012.10.004 Tirpak, F. et al. (2017) Low taurine concentrations possitively affect rabbit spermatozoa properties in later time intervals. Journal of Microbiology, Biotechnology and Food Sciences , 7, 128-131. https://doi.org/10.15414/jmbfs.2017.7.2.128-131

The blood luteinizing hormone (LH) surge in cows is well studied. However, little is known about urinary LH in cows. This study examined urinary LH concentrations after administration of gonadotropin-releasing hormone (GnRH) in six Japanese black cows to induce LH secretion from the pituitary gland into the bloodstream. Abrupt rises in plasma and urinary LH were observed after GnRH administration. Plasma and urinary LH peaked at 2 and 5 hr, respectively. A positive correlation was observed between plasma LH concentrations and urinary LH amounts. Ovulation was confirmed in the cows after 48 hr of GnRH administration. These data strongly suggest that urinary LH is derived from plasma LH, which triggers ovulation in cows.

Antioxidants have multiple protective roles in a variety of cells and thus can be used to protect sperm against cryo-damage during freezing, which affects fertility. The antioxidant resveratrol (3,5,4-trihydroxytrans-stilbene; RSV) has been reported to protect the animal sperm during cryopreservation, including human sperm. In this study, we assessed the protective effects of RSV supplementation on dog sperm cryopreservation. Semen was collected from four dogs and the effect of different concentrations of RSV (0, 100, 200, and 400 µM) on post-thaw sperm quality was examined. After thawing, sperm motility was assessed using computer-aided sperm analysis, and the structural integrity of the plasma membrane, acrosome, and chromatin was examined. In addition, their mitochondrial activity and gene expression were also assessed. Dog sperm cryopreserved with 200 µM RSV showed significant improvement in post-thaw sperm motility and viability compared with that of the control group (P

The article presents the results of a study of the efficiency of transcervical embryo transplantation obtained from donor- cows in vivo. The aim of the experiment was to compare the effectiveness of ipsilateral transfers randomized laterally from the uterine horn of embryos of different stages of development (blastocysts and morulas). The embryos were located in the left or right horn of the uterus of recipient heifers (n = 283 cows). It was experimentally established that the functional asymmetry of the ovaries of the recipients affects the level of survival of preimplantation embryos. Thus, in case of transfer to the left horn of the uterus of the recipients blastocysts 60.91% of pregnant heifers were found; morulas - 45.45%. In case of similar procedures in the right horn of the uterus - were found respectively 58.78% and 14.29% pregnant. The obtained results indicate the need for further scientific research to study the structural and physiological features that affect the survival rate of pre-implantation embryos. Key words: donor cows, recipient heifers, embryo transfer, blastocysts, morulas, transferable embryos, ipsilateral localization. References Anderson, G. B. (1983). Embryo transfer in domestic animals. Adv.Vet. Sci, 27, 129-162. Bollman, M., Greenhawk, A., Shipley, A., & Gibbons, P. (2020). Ovarian profile and pregnancy rates following ovulation synchronization and timed-artificial insemination in dairy cows. College of Veterinary Medicine. Lincoln M.U., 1291-1300. Buhrov, O. D., & Tkachova, I. V. (2013). Znachennia metodu transplantatsii embrioniv u systemi selektsiinoi roboty z maloplodnymy vydamy tvaryn. Nauk.–tekhn. Biul, 113, 43-52 (in Ukrainian). Danchuk, O. V., Karposvkii, V. I., Tomchuk, V. A. et al. (2020). Temperament in Cattle: A Method of Evaluation and Main Characteristics. Neurophysiology, 52, 73–79. https://doi.org/10.1007/s11062-020-09853-6 Does, C., Stringfellow, D., & Shultz, P. D (1984). Incorporation of follicle stimulating hormone used for embryotransfer in cattle into maltilamellar lyposomes. Theriogenology, 21(4), 661-675. https://doi.org/10.1016/0093-691X(84)90450-3 Dovhopol, V. F., Duvanov, O. V., & Ivanchenko, M. I. (2010). Efektyvnist biotekhnolohii transplantatsii embrioniv velykoi rohatoi khudoby u Poltavskii oblasti. Veterynarna medytsyna, 13, 138-141 (in Ukrainian). Duvanov, O. V. (2011). Morfolohichna kharakterystyka zhovtykh til yaiechnykiv i efektyvnist stymuliatsii poliovuliatsii koriv-donoriv. Rozvedennia i henetyka tvaryn: mizh vid. temat.nauk.zb. – K.: Ahrarna nauka, 45, 63-67 (in Ukrainian). FAO. 2010a.

The effect of estrus induction by cabergoline on gonadotropin and steroid hormone responses was examined in anestrous bitches. Eleven beagles were used in the study; seven were included in the estrus induction group and four were included in the spontaneous estrus group. Cabergoline was orally administered to the estrus induction group at 5 µg/kg once daily for four weeks, or until hemorrhagic discharge was detected. The inter-estrus interval in the estrus induction group was significantly shorter than the previous estrus interval. Bitches that showed proestrus within four weeks of treatment showed increased luteinizing hormone (LH) pulse frequency and, subsequently, increased estradiol (E2) levels. Prolactin (PRL) levels declined promptly after treatment, except in one bitch that did not show proestrus during the cabergoline treatment period. There was a significant correlation between the time to proestrus induction and the reduction in PRL levels. A positive correlation was found between the LH levels two weeks after cabergoline administration and PRL reduction. This study demonstrates that an abrupt reduction in PRL is likely to be important for initiation of estrus in bitches. A reduction in PRL indirectly leads to an increase in LH pulse frequency, which regulates follicular development in bitches. However, if the period from the end of the previous estrus to the cabergoline treatment is short, it may take some time to show proestrus without increasing E2 levels, even if the LH level increases after cabergoline administration.

Ovarian cysts are one of the most common ovarian dysfunctions in dairy cattle, which can lead to a considerable economic loss through its high incidence and can reduce the reproductive performance. Anestrus is the most significant clinical sign observed in dairy cattle suffering from this condition. For diagnosis, most of the veterinarians use a combination of methods, from ultrasonography as an additional approach to trans-rectal palpation and symptomatology when assessing ovarian cysts. Although the Ovsynch treatment seems to be preferred, the pregnancy rate after this treatment is relatively low. Despite such reports on dairy cattle ovarian cysts, the data is insufficient to validate all the characteristics of this condition. This review summarizes what the literature has so far provided from definition to treatment of ovarian cysts in dairy cattle.

Cow fertility decreases with age, but the hypothalamic pathomechanisms are not understood. Anti-Müllerian hormone (AMH) stimulates gonadotropin-releasing hormone (GnRH) neurons via AMH receptor type 2 (AMHR2), and most GnRH neurons in the preoptic area (POA), arcuate nucleus (ARC), and median eminence (ME) express AMH and AMHR2. Therefore, we hypothesized that both protein amounts would differ in the anterior hypothalamus (containing the POA) and posterior hypothalamus (containing the ARC and ME) between young post-pubertal heifers and old cows. Western blot analysis showed lower (P

This study aimed to describe the duration of inflammation after intrauterine infusion of polyvinylpyrrolidone-iodine (povidone-iodine, PVP-I), determine the effect of PVP-I infusion on the subsequent fertility, and evaluate the histopathology of the endometrium in dairy cows. In Experiment 1, 120 lactating clinically healthy Holstein-Friesian cows at 5 weeks postpartum (W5) were equally divided into three groups: intrauterine infusion of 2% PVP-I (PVP), saline (SAL), and no treatment (NTX). Endometrial cytology was performed daily from D0 (W5) to D7 to determine the percentage of polymorphonuclear cells (PMN%) in 44 of the 120 cows. All cows received timed artificial insemination at D17. In Experiment 2, 25 cows were randomly classified into sacrifice at 24 hr or 48 hr after 2% PVP-I infusion (PVP24 and PVP48), and 24, 48, 72, or 96 hr after SAL infusion (SAL24; SAL48; SAL72; SAL96), or no treatment (NTX). Histopathology was performed on the uterus of each cow. In Experiment 1, PMN% was greater in PVP (P

A 3.5-year-old, 2.9 kg, multiparous Chihuahua presented with abdominal distension; pregnancy was diagnosed. On Day 7 before parturition, prenatal sonograms showed anechoic bilateral dilated cerebral lateral ventricles, suggesting fluid-filled regions (ventriculomegaly) in one foetus. A Caesarean section was performed and the male newborn had an abnormally enlarged dome-shaped head and a cleft lip, and died 6 days after birth. According to the family pedigree, the X-linked recessive inheritance of an orofacial cleft from the unaffected mother was suggested. This report clearly demonstrates that canine foetal ventriculomegaly (hydrocephalus) can be diagnosed in utero. For dog breeds predisposed to congenital ventriculomegaly, early detection is important for the prediction of perinatal survival and adequate supportive care can be applied at delivery.

Background Domestic yaks are the most important livestock species on the Qinghai-Tibetan Plateau. Adult female yaks normally breed in the warm season (July to September) and enter anestrous in the cold season (November to April). Nevertheless, it is unclear how ovarian activity is regulated at the molecular level. Objectives The peculiarities of yak reproduction were assessed to explore the molecular mechanism of postpartum anestrus ovaries in yaks after pregnancy and parturition. Methods Sixty female yaks with calves were observed under natural grazing in Haiyan County, Qinghai Province. Three yak ovaries in pregnancy and postpartum anestrus were collected. RNA sequencing and quantitative proteomics were employed to analyze the pregnancy and postpartum ovaries after hypothermia to identify the genes and proteins related to the postpartum ovarian cycle. Results The results revealed 841 differentially expressed genes during the postpartum hypoestrus cycle; 347 were up-regulated and 494 genes were down-regulated. Fifty-seven differential proteins were screened: 38 were up-regulated and 19 were down-regulated. The differential genes and proteins were related to the yak reproduction process, rhythm process, progesterone-mediated oocyte maturation, PI3K/AKT signaling pathway, and MAPK signaling pathway categories. Conclusions Transcriptome and proteomic sequencing approaches were used to investigate postpartum anestrus and pregnancy ovaries in yaks. The results confirmed that BHLHE40, SF1IX1, FBPX1, HSPCA, LHCGR, BMP15, and ET-1R could affect postpartum hypoestrus and control the state of estrus.

Immunohistochemical investigations of the expression of surfactant protein A (SP-A) and surfactant protein D (SP-D) in the uterine and placental tissues of 13 pregnant mares were performed using anti-horse monoclonal primary antibodies. Strong positive reactions for both SP-A and SP-D were observed in the trophoblasts in the microcotyledons of the placentae at 182 to 314 days of gestation; in uterine glandular epithelial cells, faint-to-weak reactions were observed during gestation. This study describes, for the first time, the changes in the SP-A and SP-D expression levels in the endometrium of mares during gestation; the SP-A and SP-D expression levels increased after the second trimester of gestation.

History A 2-year-old sexually intact male Vietnamese potbellied pig that weighed 113 kg (249 lb) was admitted and examined to determine the cause of inappetance, irritability, and an enlargement in the inguinal region. The clinical signs had become more evident and the pig’s condition had gradually worsened during the preceding week. No treatment had been initiated prior to our examination. The owners reported that the boar would attempt to mate with a piece of polyvinyl chloride pipe. Physical examination revealed massive distention (40 X 25 cm [15.7 X 9.8 inches]) in the inguinal region and caudoventral portion of the abdomen. Palpation of the enlargement revealed it was firm and excessively warm but could be compressed, which suggested it was filled with fluid. The prepuce was edematous. There was evi dence of irritation of the skin over the right inguinal region. Question

Inflammation and fibroproliferative diseases may be modulated by epigenetic changes. Therefore, we suggest that epigenetic mechanisms could be involved in equine endometrosis pathogenesis. DNA methylation is one of the methods to evaluate epigenetics, through the transcription of methyltransferases (DNMT1, DNMT3A, DNMT3B). The correlation between DNMTs and collagen (COL) transcripts was assessed for the different Kenney and Doig's (Current Therapy in Theriogenology. Philadelphia: WB Saunders; 1986) endometrium categories. Endometrial biopsies were randomly collected from cyclic mares. Histological classification (category I, n = 13; II A, n = 17; II B, n = 12; and III, n = 7) and evaluation of COL1A2, COL3A1 and DNMTs transcripts by qPCR, were performed. Data were analysed by one-way analysis of variance (ANOVA), Kruskal-Wallis test and Pearson correlation. As mares aged, there was an increase in endometrium fibrosis (p < .01), and in DNMT1 mRNA (p < .001). Considering DNMT3B transcripts for each category, there was an increase with fibrosis (p < .05). No changes were observed for DNMT1 and DNMT3A transcripts. However, DNMT3A mRNA levels were the highest in all categories (p < .01). In category I endometrium, a positive correlation was observed for transcripts of all DNMTs in both COLs (p < .01). In category IIA, this correlation was also maintained for all DNMTs transcripts in COL1A2 (p < .05), but only for DNMT3B in COL3A1 (p < .05). In category IIB, there was a positive correlation between DNMT3B and COL3A1 (p < .05). In category III, a positive correlation was only observed between DNMT3B and COL3A1 (p < .05). Our results suggest that there is a disturbance in COLs and DNMTs correlation during fibrosis.

Postpartum uterine disease due to poor uterine involution continues to be a significant factor that contributes to poor reproductive efficiency in dairy cattle. Therapy that increases the frequency, duration and strength of uterine contractions in the postpartum period might enhance uterine involution, resulting in improved reproductive performance. The objective of this clinical trial was to study the effect of two uterine ecbolic therapies, oxytocin and prostaglandinF2α on uterine involution, postpartum endometritis, and reproductive performance. A randomized double-blinded clinical trial was conducted in 118 dairy cows from two research herds that had normal parturition and expulsion of the fetal membranes. Within 24 hr after calving, cows were randomly assigned to receive intramuscular injections twice a day of 50 IU of oxytocin, or 25 mg of dinoprost (PGF2α) or saline (control) for 7 days. Cows were monitored from day 3 to day 63 ± 3 days postpartum by transrectal palpation of the uterus, vaginoscopy, Metricheck® examination and by endometrial cytology. Blood samples were collected for measurement of progesterone in weeks 3 (21 ± 3 days), 5 (35 ± 3 days), 7 (49 ± 3 days), and 9 (63 ± 3 days) postpartum. Herd breeding records were obtained to determine reproductive performance in all cows. Neither oxytocin nor prostaglandin F2α therapy during the first week postpartum had any significant effect on the rate of uterine involution, prevalence of endometritis or reproductive performance, compared to untreated controls. Ecbolic drugs, as used here, are not recommended for use in clinical practice to improve involution or reproductive tract health in normal cows.

We investigated the effects of genetic background on the responses to superovulation in Japanese Black cattle. The genotype frequencies of GRIA1 and FSHR relating to ovulation and follicular development in each of the major bloodlines—Tajiri, Fujiyoshi, and Kedaka—were analyzed. The Tajiri line had the lowest frequency of G allele homozygosity of c.710A>G in GRIA1 among the three bloodlines, and deviation from Hardy–Weinberg equilibrium was detected. Genotype frequencies of c.337C>G, c.871A>G, and c.1973C>G in FSHR were in Hardy–Weinberg equilibrium in all bloodlines. The results of generalized linear mixed-model analyses showed that farm, levels of plasma anti-Müllerian hormone (AMH) concentration, age in months, repeated superovulation, c.337C>G in FSHR, and bloodlines had significant effects on the responses to superovulation. The number of transferable embryos in the group heterozygous for c.337C>G in FSHR was significantly higher than that in the group homozygous for the C allele. The Kedaka line showed a significantly higher number of ova/embryos, fertilized embryos, and transferable embryos than the Tajiri and Fujiyoshi lines. The concentration of circulating AMH is a useful endocrine marker for antral follicle counts. This study revealed the effects of genetic background on the responses to superovulation using levels of plasma AMH concentration as a covariate. The prominent effect of genetic background on superovulation in the Kedaka line requires additional studies to confirm the genomic regions and polymorphisms that are involved in the trait.

The present study aimed to determine possible risk factors affecting reproductive performance, especially time taken to establish pregnancy in dairy cows, using Cox's proportional hazard model. The data were collected from 154 Holstein Friesian cows (199 lactations). Cows diagnosed with a vaginal discharge score (VDS) of one or calving abnormality showed significantly delayed pregnancy: hazard ratio (HR)=0.654 (95% confidence interval [CI]: 0.436-0.983; P=0.041) and HR=0.457 (95% CI: 0.270-0.774; P=0.004), respectively. Our study suggested that the occurrence of VDS of one or calving abnormality might be possible risk factors increasing the number of days open and affecting reproductive performance in dairy cows.

In this study, we aimed to elucidate the utility of transrectal ultrasonography (TRUS) and blood lactate concentration (bLac) measurement to diagnose cows with severe uterine torsion. We investigated the association of TRUS and bLac measurement with macroscopic findings on laparotomy for severe uterine torsion in nine cows. We found that an increased ultrasonographic cross-sectional thickness (15-25 mm) and multiple hypoechogenic areas corresponded to macroscopic vascular compromise in the uterus on laparotomy. In addition, bLac was elevated (≥5.0 mmol/l) in cows showing uterine necrosis on laparotomy. A combined diagnostic approach with TRUS and bLac measurement enables assessment of the uterine vascular status and has utility for selecting the treatment option (including laparotomy) and predicting the outcomes.

A routine parentage test as part of a conservation program for Kiso horses identified a possible sex chromosome anomaly in a 7 months-old filly because of an aberrant result using LEX3, an X-linked marker. We then analyzed X-linked markers (LEX26, TKY38, and TKY270), Y-linked markers (Eca.YH12, Eca.YM2, Eca.YA16, and the sex-determining region Y gene), and an X/Y marker (Amelogenin gene). This analysis demonstrated that the filly had not inherited an X chromosome from her sire. A karyotyping analysis confirmed that the filly was 63,XO. As it was suspected that the horse would be sterile, we avoided using the horse as a broodmare; the information should also serve to prevent unnecessary conflict between owners transferring and receiving the horse.

The purpose of this study was to clarify the therapeutic effects of oxytetracycline (OTC) as a first-line antibiotic in cattle with acute Escherichia coli mastitis and systemic signs. Drug susceptibility was determined by the minimum inhibitory concentration (MIC) of E. coli isolated from cows with acute E. coli mastitis (n=38). Cattle were divided into OTC-susceptible (S, n=30) and OTC-resistant (R, n=8) groups. They were further subdivided according to susceptibility to the antibiotic used as a second treatment, into susceptible-susceptible (SS, n=30), resistant-susceptible (RS, n=5), and resistant-resistant (RR, n=3) groups. Clinical signs on the day after initial treatment were compared between S and R groups as short-term indicators of treatment effects. The 28-day survival rate of cattle was then compared among SS, RS, and RR groups as a long-term indicator of treatment effects. There were no differences in clinical signs between S and R groups on the day after the first dose, but the 28-day survival rate was significantly greater in the SS group than in the RR group (P=0.04). The results demonstrated that an effective drug is essential for first-line treatment of acute coliform mastitis. However, anticipating the effectiveness of a first-line antibiotic based on clinical symptoms at the second day of treatment is impossible. It is important to build a picture of drug resistance trends in cattle herds for empirical selection of antibiotics to be administered.

The objective of this study was to evaluate the breeding performance of Indonesian beef cattle (Ongole cross) as recipients for embryo transfer using Limousin embryos. As a result, the pregnancy rate was 35% (7 out of 20 cows). There was a significant difference (P

This study investigated whether treatment with the mitogen-activated protein kinase kinase inhibitor U0126 during in vitro maturation (IVM), which has previously been reported to improve oocyte developmental competence, is practical for use in calf production using ovum pick up (OPU)-derived oocytes. Two Japanese Black cows were repeatedly and simultaneously treated to stimulate follicular growth and were prepared for OPU. Cumulus-oocyte complexes (COCs) were collected from one cow using a collection medium containing 5 µM U0126 and were cultured in medium supplemented with the same concentration of U0126 for the first 2 hr of IVM; COCs from the other cow were used as controls without U0126 treatment. The cows were exchanged between the two groups at every sequential OPU (n=8). The number of oocytes developing to blastocysts in the U0126-treated group (39.1%, 34/87) was significantly higher than that in the control group (22.1%, 19/86). Eight blastocysts produced with U0126 treatment were transferred to recipients, and four normal calves were obtained. The results indicate that embryos develop efficiently from OPU-derived oocytes treated with U0126, and that these embryos may be of practical use in calf production.

Background: Sperm capacitation is a complex process involving several changes in the spermatozoa, allowing to acquire the capacity to fertilize the oocyte: reorganization of the plasmalemma, protein tyrosine phosphorylation, and motility hyperactivation. Cytosolic Ca2+ concentration plays an important role in these events. In addition, melatonin is important for the functionality of the reproductive tract of mammals. In males, melatonin has a direct influence on testosterone levels and sperm quality (Li and Zhou, 2015, Clin Chim Acta 446:175–80), and it could regulate sperm capacitation, maybe by modulating ROS levels or by binding to its spermatic membrane receptors MT1 and MT2 (Gonzalez-Arto et al., 2016, Theriogenology 86:1958–68).

Oxidative stress owing to an imbalance between reactive oxygen species and antioxidants, such as coenzyme Q10 (CoQ10), is a major contributor to male infertility. We investigated the effects of the reduced form of CoQ10 (ubiquinol) supplementation on semen quality in dogs with poor semen quality. Three dogs received 100 mg of ubiquinol orally once daily for 12 weeks. Semen quality, serum testosterone, and seminal plasma superoxide dismutase (SOD) activity were examined at 2-week intervals from 2 weeks before ubiquinol supplementation to 4 weeks after the treatment. Ubiquinol improved sperm motility, reduced morphologically abnormal sperm, and increased seminal plasma SOD activity; however, it had no effect on testosterone level, semen volume, and sperm number. Ubiquinol supplementation could be used as a non-endocrine therapy for infertile dogs.

The factors that affect the interval to ovulation, the type of ovulated dominant follicle (DF), and the cause of anovulation after prostaglandin (PG) treatment were investigated. Nine cows were assigned to six groups (54 cows in total) but the group size was later fixed at eight cows (48 in total). They received 25 mg tromethamine dinoprost as dinoprost on Day 6 (Group D6), Day 7 (Group D7), Day 8 (Group D8), Day 9 (Group D9), Day 10 (Group D10), or Day 11 (Group D11) after natural ovulation (Day 0). If the DF did not ovulate, then the cow was assigned to Group NO. In Group D6, the 1st DF ovulated in all cows 4 days after PG treatment, whereas in Groups D9, D10, and D11, the 2nd DF ovulated in all cows 4 to 7 days after PG treatment. In 10 cows, the DF did not ovulate, and late anovulation was significantly higher in Group D6 cows than in Group D11 cows. The progesterone (P4) levels decreased to less than 1 ng/ml in all groups on the day after PG treatment. The estradiol-17β (E2) levels began to increase after PG treatment and peaked at 2 days before ovulation in the cows that ovulated. In anovulated cows, E2 tended to be higher and there was no clear E2 peak in some cows. These results indicated that the number of days to ovulation, the type of ovulated DF, and anovulation were affected by factors that were associated with the DF when it was producing E2.

Made available in DSpace on 2020-12-12T01:58:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-01-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Universidade Estadual Paulista In human neonates, when the umbilical cord is kept intact postpartum, blood continues to flow to the neonate, but this procedure might be difficult in dogs owing to a shorter umbilical cord and several neonates in a litter. However, it might be possible to detach the placenta and keep the umbilical cord intact, allowing residual blood to flow to the puppies. This study compared the effects of clamping versus no clamping of the umbilical cord in dogs born by cesarean section on neonatal vitality. The puppies were assessed by Apgar and reflex scores. Fifty puppies delivered from 16 bitches were randomly allocated to receive immediate umbilical cord clamping (n=25) or no clamping for at least 3 min after the onset of breathing (n=25). The puppies were assessed during the first 5 min of life and 10 min after the first assessment. The no clamping group showed significantly higher Apgar scores (second assessment, P

Serum anti-Mullerian hormone (AMH), a marker of equine cryptorchidism, is detectable in intact and cryptorchid stallions but not in geldings because it is secreted from Sertoli cells. A 4-year-old uncastrated Thoroughbred racehorse had no visible testes; therefore, the horse was considered a bilateral cryptorchidism. However, the serum AMH was undetectable (

The selection of competent oocytes to completion of meiosis in vitro, fertilization or reconstructing (cloning, transgenesis) is the initial stage of cell reproductive technologies in animal husbandry. The development of effective methods of early prediction prospective potencies for extracorporeal maturation and fertilization of oocyte is the actual problem of rapidly developing embryo technologies. Numerous factors determined developmental competence of the oocytes. Brilliant cresyl blue (BCB) staining has been used for selection of oocytes from several mammalian species, including pigs (Ericsson S. et al, Theriogenology, 39(1): p.214, 1993). BCB determines the intracellular activity of glucose-6-phosphate dehydrogenase, which plays an important role in cell growth, as a key enzyme in the pentose phosphate cycle. The enzyme activity in the growing oocyte increases, opposite in the oocytes that have finished growth phase it decreases (Alm et al., 2005). BCB - diagnostics of the initial population of oocytes based on staining with vital dye brilliant cresyl blue have proposed as an effective indicator of completion of oocyte growth phase. The aim of the present study was to evaluate the developmental competence of porcine oocytes that have finished growth phase (BCB+) in vivo depending on diameter (d) of follicles (d

BNIP3 (BCL2/adenovirus E1B nineteen kilodalton interacting protein-3), a member of the BCL2 family, is activated under hypoxic conditions and induces apoptosis or mitochondrial autophagy for adapting cells to hypoxia. The physiological roles of BNIP3 in the mammalian ovary are still unclear. In order to understand the role of BNIP3 in the bovine ovary, we examined its mRNA and protein expressions of BNIP3 in follicular granulosa cells and corpus luteum (CL). BNIP3 mRNA and protein expressions in granulosa cells from large follicles (>10 mm) at the follicular stage were much higher than those in small follicles (2–8 mm). BNIP3 mRNA and protein expressions in the CL peaked at the early luteal stage. In bovine granulosa cells cultured for 6 hr under hypoxia (3% O2) and normoxia (20% O2), BNIP3 mRNA expression was higher under hypoxia. These results of the present study suggest that BNIP3 has some roles in luteal formation in the bovine ovary, and that the highly expressed BNIP3 in the granulosa cells from large follicles at the follicular stage is related to the roles of BNIP3 in the luteal formation.

The objective of this study was to evaluate the effects of post artificial insemination (AI) treatment with intravaginal progesterone device (P4 device) on conception rate, synchronization of returning estrus and plasma P4 concentration in Japanese Black cows. Nineteen cows were treated with DIB (1.0 g P4) from Day 12 to 19 (Day 0=day of the first AI), 27 cows were treated with a CIDR (1.9 g P4) from Day 12 to 19, and 33 cows were not treated after the first AI (control). Estrous behavior was daily examined between Day 20 and 25, and cows returning to estrus were inseminated (the second AI). On Day 19, plasma P4 concentration was not different among DIB, CIDR and control groups. There was no significant difference in conception rate after the first AI among three groups (DIB: 63.2%, CIDR: 66.7% and control: 72.7%). In non-pregnant cows, there was no significant difference in the proportion of cows showed returning estrus between Day 20 and 25 (DIB: 57.1%, CIDR: 22.2% and control: 44.4%), and day of returning estrus was not synchronized. The overall conception rate after the first and second AI was not different among the groups. In conclusion, post-AI treatment with intravaginal devices containing 1.0 and 1.9 g P4 from Day 12 to 19 neither increased plasma P4 concentration nor improved fertility and synchronization of the returning estrus in Japanese Black cows.

Amniotic mesenchymal stem cells (AMSCs) from livestock are valuable resources for animal reproduction and veterinary therapeutic. The purpose of this study is to explore a suitable way to isolate and culture the buffalo AMSCs (bAMSCs), and to identify their biological characteristics. Digestion with a combination of trypsin-EDTA and collagenase type I could obtain pure bAMSCs more effectively than trypsin-EDTA or collagenase type I alone. bAMSCs could proliferate steadily in vitro culture and exhibited fibroblastic-like morphology in vortex-shaped colony. bAMSCs were positive for MSC-specific markers CD44, CD90, CD105, CD73, β-integrin (CD29) and CD166, and pluripotent markers OCT4, SOX2, NANOG, REX-1, SSEA-1, SSEA-4 and TRA-1-81, but negative for hematopoietic markers CD34, CD45 and epithelial cells specific marker Cytokeratin 18. In addition, bAMSCs were capable of differentiating into adipogenic, osteogenic, chondrogenic and neural lineages, with expression of FABP4, Ost, ACAN, COL2A1, Nestin and β III-tubulin. Glycogen synthase kinase 3 inhibitor: kenpaullone promoted bAMSCs to differentiate into neural lineage. This study provides an effective protocol to obtain and characterize bAMSCs, which have proven useful as a cell resource for buffalo cell reprogramming studies and transgenic animal production.

Article Details: Received: 2018-04-19 | Accepted: 2018-06-27 | Available online: 2018-06-30 https://doi.org/10.15414/afz.2018.21.02.52-59 The aim of this work was to describe the microscopic and submicroscopic changes of the uterus of 20 pinzugus heifers in their postnatal development (12 th , 24 th and 36 th week of age) and in the experimental form of subclinical hypoglycemia. Uterine specimens were obtained from the uterine horns by a vivisection for histological studies. Samples were fixed for light microscopy (LM) in formaldehyde and for scanning by electron microscope (SEM) and transmissive electron microscopy (TEM) in glutaraldehyde. Subsequently, the samples were processed in the usual manner as used in the LM and electron microscopic studies laboratories. The uterus increased to 24 th weeks by 19.4 %. The uterine weight decreased significantly (P

The treatment of cystic ovarian disease (COD) in dairy cows is still controversial, and some researchers recommend using gonadotropin-releasing hormone (GnRH) regardless of the type of cysts. The aim of this study was to comparatively evaluate the reproductive performance of cows diagnosed with follicular or luteal cystic structures, after treatment with either buserelin acetate (GnRH agonist) or dinoprost (prostaglandin F2-alpha or PGF2α). The diagnosis was established by ultrasonographic examinations performed twice a month starting 40–45 days after calving, until the cows were diagnosed pregnant after artificial insemination. Both types of cysts were treated either with 21 µg buserelin acetate or 25 mg dinoprost, resulting in four subject groups. After treatment, the estrus rate in cows with follicular cysts treated with dinoprost (55.1%) was significantly lower (P

A major role of the corpus luteum (CL) is to produce progesterone (P4). The CL has immature vasculature shortly after ovulation, suggesting it exists under hypoxic conditions. Hypoxia-inducible factor-1 (HIF1) induces the expression of glucose transporter 1 (GLUT1). To clarify the physiological roles of GLUT1 in bovine CL, we examined GLUT1 mRNA expression in the CL under hypoxic conditions by quantitative RT-PCR. We also measured the effects of glucose (0–25 mM) and GLUT1 inhibitors (cytochalasin B, STF-31) on P4 production in bovine luteal cells. GLUT1 mRNA expression in bovine CL was higher at the early luteal stage compared to the other later stages. Hypoxia (3% O2) increased GLUT1 mRNA expression in early luteal cells, but not in mid luteal cells. Glucose (0–25 mM) increased P4 production in early luteal cells, but not in mid luteal cells. Both GLUT1 inhibitors decreased P4 production in early and mid luteal cells. Overall, the results suggest that GLUT1 (possibly induced by hypoxic conditions in the early CL) plays a role in the establishment and development of bovine CL, especially in supporting luteal P4 synthesis at the early luteal stage.

In order to determine blood lactate concentrations (bLac) and their validity as a diagnostic marker in bovine uterine torsion, blood samples were taken from 54 Holstein cows with uterine torsion before the correction of torsion. bLac in a group of cows with and without uterine necrosis were 15.0 and 3.0 mmol/l, respectively (P5.0 and >6.5 mmol/l, respectively. These findings suggest that in dairy cows with uterine torsion, an increase in bLac is a diagnostic predictor of uterine necrosis as well as poor prognosis in dams.

We aimed to determine gene expression patterns in the anterior pituitary (AP) of heifers before and after ovulation via deep sequencing of the transcriptome (RNA-seq) to identify new genes and clarify important pathways. Heifers were slaughtered on the estrus day (pre-ovulation; n=5) or 3 days after ovulation (post-ovulation; n=5) for AP collection. We randomly selected 4 pre-ovulation and 4 post-ovulation APs, and the ribosomal RNA-depleted poly (A)+RNA were prepared to assemble next-generation sequencing libraries. The bovine APs expressed 12,769 annotated genes at pre- or post-ovulation. The sum of the reads per kilobase of exon model per million mapped reads (RPKM) values of all transcriptomes were 599,676 ± 38,913 and 668,209 ± 23,690, and 32.2 ± 2.6% and 44.0 ± 4.4% of these corresponded to the AP hormones in the APs of pre- and post-ovulation heifers, respectively. The bovine AP showed differential expression of 396 genes (P

Uridine 5’-diphospho-glucuronosyltransferase (UGT) liver activity was measured using estradiol-17β as a substrate in dairy cows with follicular cysts. The activity was significantly lower than that in dairy cows with normal estrous cycles (P

We examined the effect of human chorionic gonadotropin (hCG) treatment on double ovulation rate of first-wave follicles five days after estrus in lactating dairy cows. Cows were divided into two groups: 1) Ipsilateral group (IG; n=35), in which the first-wave dominant follicle (DF) was ipsilateral to the corpus luteum (CL), and 2) Contralateral group (CG; n=30), in which the first-wave DF was contralateral to the CL five days after estrus, then 1,500 IU of hCG was administrated. Double ovulation rate was significantly higher in the CG (26.7%) than in the IG (2.9%). This study demonstrate that the double ovulation rate of first-wave follicles was higher in the first-wave DF located in the contralateral ovary to the CL.

application/pdf, There has been no report of equine embryo transfer in Japan for the last 24 years. Our objective was to establish an effective protocol for embryo transfer in domestic horse. A Hokkaido native pony was bred by deep-horn artificial insemination with frozen semen from a Connemara pony. Embryo collection was performed using a non-surgical method on day 7. Two embryos were obtained from three flushes (67% recovery) and were transferred fresh into crossbred recipient mares. Both recipient mares were diagnosed pregnant (100% pregnancy rate) 5 days after embryo transfer and had normal progesterone levels until the end of the observation on day 35 of gestation. This is the first successful embryo transfer in Japan by artificial insemination using frozen semen. Embryo transfer technology could be extremely useful in improving the productivity of Japanese domestic and special riding horses.

Two American Cocker Spaniels and one Bichon Frise were presented to our veterinary teaching hospital with an enlarged clitoris. Diagnostic imaging showed that the structure was composed of bony material. Exploratory laparotomy revealed uterine-like structures and testes which had an epididymis unilaterally. Surgical removal of internal genitalia, gonads and protruded clitoris were performed well. Histological evaluation revealed; inactive testes, female internal genital tracts with ambisexual ductal remnants, and prominent ossification in the clitoris. All 3 cases were diagnosed with male pseudohermaphroditism. In author's knowledge, this is the first report in Bichon Frise dog with os clitoris and also, it describes not common cases in small dog breeds with os clitoris.