In Vitro Embryos of Romosinuano and Tropical Milking Cattle during Three Seasons in Veracruz, Mexico.

One of the main factors that influences the fertility of cattle in grazing systems in hot tropical climates is heat stress. The objective of this study was to evaluate the effect of season, breed, hormonal and physiological condition on the quantity and quality of cumulus-oocyte complexes (COCs) and embryos produced in vitro, from Romosinuano (RM) and Tropical Milking (TM) donors. Three ovum pick-up and in vitro fertilization (OPU-IVF) were performed, one per season: hot dry (HD; 10, 10), hot humid (HH; 9, 9) and fresh dry (FD; 7, 10) in RM and LT donors. Serum levels of cortisol, insulin and glucose were measured, in addition to heart rate (HR), respiratory rate (RR) and rectal temperature (RT). Effect of season x genotype interaction (p ≤ 0.05) was observed in all COC variables and only in cleavage embryos (CLI) (p ≤ 0.05). Body weight (BW) affected all COC variables (p ≤ 0.01), except unviable (UNV) although affected degenerated embryos (DEG) (p ≤ 0.01) and total blastocysts (BLAST) (p ≤ 0.01). Cow age only affected viable COCs (VIAB) (p ≤ 0.05), code one blastocysts (BC1) and BLAST (p ≤ 0.01). Cortisol affected total COCs (COCsT), VIAB and total matured in vitro (TMIV) (p ≤ 0.01), as well as CLI, BC1 (p ≤ 0.01) and BLAST (p ≤ 0.05). Insulin affected COCsT (p ≤ 0.01), UNV (p ≤ 0.05), denuded oocytes (DE) (p ≤ 0.01), BC1 and code two blastocysts (BC2) (p ≤ 0.01). Glucose affected all COC variables (p ≤ 0.01), except UNV and all embryo variables except BC2. HR affected COCsT, DE, TMIV (p ≤ 0.01), CLI, BLAST and DEG (p ≤ 0.05). RR affected COCsT, UNV, VIAB, CLI (p ≤ 0.05), BC1, BLAST and DEG (p ≤ 0.01). RT only affected DE, VIAB (p ≤ 0.01) and BLAST (p ≤ 0.05). The seasonal climatic year variation of Veracruz and changes in physiological and hormonal variables have diverse effects on the cumulus-oocyte complexes and embryos produced by RM and TM donors.

Resveratrol Reduces ROS by Increasing GSH in Vitrified Sheep Embryos.

The in vitro production and cryopreservation of mammalian embryos generates reactive oxygen species (ROS) due to conditions of the system that can overcome their antioxidant protection. Resveratrol is an antioxidant used in in vitro systems to improve blastocyst rates, but its effect on antioxidant enzymes such as glutathione (GSH) in embryos produced by in vitro fertilization (IVF) after vitrification has not been reported. The objective of this study was to evaluate the effects of resveratrol on the in vitro maturation medium (IVM) of sheep oocytes (Ovis aries) on the levels of ROS and GSH in embryos produced by IVF subjected to vitrification. Resveratrol was added at 0 µM, 0.25 µM, 0.5 µM, and 1 µM during oocyte in vitro maturation (IVM). Matured oocytes were fertilized with thawed ram sperm. Embryos were cultured in sequential media until blastocysts, were then vitrified for 24 h, and, after heating, they were stained with DCFH-DA (2',7'-dichlorodihydrofluorescein diacetate) to determine the presence of ROS and with Cell Tracker Blue® for the presence of GSH. The quantitative values of ROS and GSH were obtained through the Image J image processor. The results showed that resveratrol increased GSH and decreased ROS production (p < 0.05) in a dose-dependent manner. It is concluded that its use in sheep oocytes during IVM has a beneficial effect on embryos produced by IVF subjected to vitrification by decreasing reactive oxygen species levels and facilitating the generation of embryo antioxidant enzymes like glutathione.

Reproduction of Sheep through Nuclear Transfer of Somatic Cells: A Bibliometric Approach.

Somatic cell nuclear transfer (SCNT) is a reproductive biotechnology with great potential in the reproduction of different species of zootechnical interest, including sheep. This study aimed to carry out a bibliometric analysis of scientific papers published on the application of SCNT in sheep reproduction during the period 1997-2023. The search involved the Science Citation Index Expanded and Social Sciences Citation Index databases of the main collection of the Web of Sciences with different descriptors. A total of 124 scientific papers were analyzed for different bibliometric indicators using the VOSviewer software. Since 2001, the number of SCNT-related papers that have been published concerning sheep reproduction has increased and it has fluctuated in ensuing years. The main authors, research groups, institutions, countries, papers, and journals with the highest number of papers related to the application of SCNT in sheep reproduction were identified, as well as the topics that address the research papers according to the terms: somatic cell, embryo, oocyte, gene expression, SCNT, and sheep.

Detection of Candidate Genes Associated with Fecundity through Genome-Wide Selection Signatures of Katahdin Ewes.

One of the strategies to genetically improve reproductive traits, despite their low inheritability, has been the identification of candidate genes. Therefore, the objective of this study was to detect candidate genes associated with fecundity through the fixation index (FST) and runs of homozygosity (ROH) of selection signatures in Katahdin ewes. Productive and reproductive records from three years were used and the genotypes (OvineSNP50K) of 48 Katahdin ewes. Two groups of ewes were identified to carry out the genetic comparison: with high fecundity (1.3 ± 0.03) and with low fecundity (1.1 ± 0.06). This study shows for the first time evidence of the influence of the CNOT11, GLUD1, GRID1, MAPK8, and CCL28 genes in the fecundity of Katahdin ewes; in addition, new candidate genes were detected for fecundity that were not reported previously in ewes but that were detected for other species: ANK2 (sow), ARHGAP22 (cow and buffalo cow), GHITM (cow), HERC6 (cow), DPF2 (cow), and TRNAC-GCA (buffalo cow, bull). These new candidate genes in ewes seem to have a high expression in reproduction. Therefore, future studies are needed focused on describing the physiological basis of changes in the reproductive behavior influenced by these genes.

Kisspeptin Stimulates the Pulsatile Secretion of Luteinizing Hormone (LH) during Postpartum Anestrus in Ewes Undergoing Continuous and Restricted Suckling.

This study tested whether the intravenous application of kisspeptin can stimulate the pulsatile secretion of LH in suckling ewes during postpartum anestrus. Ten days after lambing, Pelibuey ewes were allocated among two groups: (1) continuous suckling (n = 8), where the lambs remained with their mothers; and (2) restricted suckling (n = 8), where the mothers suckled their lambs twice daily for 30 min. On Day 19 postpartum, the ewes were individually penned with ad libitum access to water and feed and given an indwelling catheter in each jugular vein. On Day 20, 4 mL of blood was sampled every 15 min from 08:00 to 20:00 h to determine LH pulse frequency. At 14:00 h, four ewes in each group received 120 µg of kisspeptin diluted in 3 mL of saline as a continuous infusion for 6 h; the remaining four ewes in each group received only saline. The interaction between kisspeptin and suckling type did not affect LH pulse frequency (p > 0.05). Before kisspeptin administration, pulse frequency was similar in all groups (1.50 ± 0.40 pulses per 6 h; p > 0.05). With the application of kisspeptin, pulse frequency increased to 3.50 ± 0.43 pulses per 6 h (p ≤ 0.014), so the concentration of LH (1.11 ± 0.14 ng mL-1) was greater in kisspeptin-treated ewes than in saline-treated ewes (0.724 ± 0.07 ng mL-1; p ≤ 0.040). The frequency of LH pulses was greater with restricted suckling than with continuous suckling (2.44 ± 0.29 versus 1.69 ± 0.29 pulses per 6 h; p ≤ 0.040). We conclude that intravenous application of kisspeptin increases the pulsatile secretion of LH in suckling ewes and that suckling might reduce kisspeptin neuronal activity, perhaps explaining the suppression of ovulation. Moreover, the effects of kisspeptin and suckling on pulsatile LH secretion appear to be independent, perhaps operating through different neural pathways.

Influence of FecGE mutation on the reproductive variables of Pelibuey ewes in the anestrus period.

The growth and differentiation factor 9 (GDF9) intervenes in the fecundity and prolificacy of the ewe, which are important variables that participate in the reproductive efficiency of a flock. The objective of this study was to evaluate the influence of FecGE mutation of the gene GDF9 in the natural response of the manifestation to estrus, return to estrus, ovulation rate, pregnancy, lambing, prolificacy, and fecundity rate in Pelibuey ewes, during the anestrus period. The sequences of the exon 2 of the gene GDF9 were obtained from blood samples collected in Whatman™ FTA™ cards from 42 multiparous Pelibuey ewes with reproductive records. For this purpose, the quality of the sequences was analyzed and the polymorphisms and genotypes were searched for. The ewes were grouped according to their group: (a) homozygous or Embrapa (GG), (b) wild (AA), and (c) group without gene (sG). All the ewes studied manifested estrus behavior, but none showed signs of return to estrus after natural mating (p > 0.05); likewise, the pregnancy and lambing rates (p > 0.05) did not show differences between groups. However, the group GG presented higher ovulation rate, prolificacy, and fecundity rate (p < 0.05), compared to groups AA and sG. Although no differences were found in the manifestation of estrus, return to estrus, and percentage of pregnancy and lambing in females from the genotypes studied, the homozygous ewes GG presented 1.22 and 1.72 more corpus luteum (CL, p < 0.05), prolificacy of 0.7 and 0.7, and fecundity rate of 0.8 and 1.0 more lambs per ewe (p < 0.05) than the ones produced by the wild-type AA and sG groups, respectively.

SEROPREVALENCE OF ANTIBODIES AGAINST MYCOBACTERIUM AVIUM SUBSP. PARATUBERCULOSIS AND ITS RELATIONSHIP TO AGE AND SEX OF TEXAS WHITE-TAILED DEER (ODOCOILEUS VIRGINIANUS) IN COAHUILA, MEXICO.

Paratuberculosis (PTB) is a disease that affects cattle (Bos taurus), goats (Capra aegagrus hircus), sheep (Ovis aries), and wild animals, such as white-tailed deer (Odocoileus virginianus), since all ruminants are susceptible. The causal agent is Mycobacterium avium subsp. paratuberculosis (MAP). The disease is chronic, consumptive, and incurable; it causes chronic granulomatous gastroenteritis with lymphangiectasis and lymphangitis leading to a syndrome of malnutrition and eventually to death. Mycobacterium avium subsp. paratuberculosis is transmitted in feces mainly orally; however, it can also be transmitted vertically. Thus, the objective of this study was to determine the seroprevalence of MAP antibodies and its relationship to age and sex of Texas white-tailed deer in the subclinical stage of PTB in Coahuila, Mexico. The entire population (n=99) belonging to the Wildlife Management and Conservation Unit (WMCU) San Juan, Monclova, Coahuila, Mexico was captured. Mycobacterium avium subsp. paratuberculosis was diagnosed using an enzyme-linked immunosorbent assay by serologic test. Seroprevalence variables of adult vs. young females and males vs. females were compared. The treatments were assigned at random. For the analysis of data, the chi-square test was used. Total seroprevalence in an intensive WMCU was 16% (16/99). Total seroprevalence by sex was 5.0% (5/99) for males and 11% (11/99) for females, and total seroprevalence by age was 7% (7/99) for young and 9% (9/99) for adult. Within sex, the seroprevalence in males was 16% (5/31) and 16% (11/68) in females. There were no statistical differences for any of the comparisons. Total seroprevalence of the white-tailed deer population in the WMCU was 16%, and PTB seroprevalence was independent of sex or age of the sampled individuals of this population.

Relation of the maximum temperature and relative humidity close to the insemination with the tropical milking criollo heifer's gestation in three seasons.

The high climatic variability of hot climates of the intertropical zone reduces cattle fertility. In dairy cows in temperates zones, the THI has been used to evaluate the temperature and relative humidity (RH) joint effect in reproduction, but its use is not recommended in all geographic zones; in hot climates, the maximum temperature (Tmax) can provide more convenient information than THI. The objective of this study was to determine the artificial insemination (AI) service seasons and their joint effect with the maximum temperature and relative humidity of the previous seven days, the service day, and posterior seven days to the AI in the tropical milking criollo (LT) heifer's gestation. Climatic data was used to define three seasons: hot-dry (HD), hot-humid (HH), and fresh-dry (FD), and 313 artificial insemination services from 176 heifers were analyzed over fourteen years. The seasons were determined by cluster analysis. Gestation at first service (GF) was analyzed with a logistic regression model and global gestation (GG) with a mixed linear generalized model. The Tmax of previous seven days insemination [Formula: see text] - 0.20 ± 0.09 (p ≤ 0.02) in HD (p ≤ 0.02) and RH of seven days posterior insemination [Formula: see text]= - 0.08 ± 0.04 (p ≤ 0.04) in HD (p ≤ 0.01) affected GF. No effect of the Tmax and RH on the service day was observed (p > 0.05). The highest GG probabilities were higher than 0.70 in HH and FD, making those seasons the most suitable for inseminating LT heifers.

Detection of Mycobacterium avium subsp. paratuberculosis in reproductive tissue and semen of naturally infected rams.

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of paratuberculosis (PTB), disease that causes a syndrome of bad nutrient absorption, weight loss and eventually death. The intestine is the main target organ where the infection develops; however, there is evidence of infection by MAP in extra-intestine sites of sheep, including mesenteric nodes and semen. The aim of the study was to identify the presence of MAP in reproductive tissue and semen of infected Pelibuey rams in clinical state of PTB. Seven rams were used in clinical PTB state and a non-infected ram by MAP of the Pelibuey breed, confirmed by serology, nPCR and bacteriological culture, with average weight and age of 57.23 ± 1.73 kg and 2.91 ± 0.17 years, respectively. The presence of MAP was identified in different tissue samples: spleen (1/7, 14.3% and 2/7, 28.6%), small intestine (3/7, 42.9% and 4/7, 57.1%) and mesenteric lymph nodes (3/7, 42.9% and 3/7, 42.9%), with nPCR and culture, respectively. It was also identified in epididymis tissue (1/7, 14.3%), Cowper gland (2/7, 28.6%) and prostate (1/7, 14.3%), using nPCR, although without detection in culture. It was identified in testicular tissue in 42.8% (3/7; culture or nPCR technique), but in 28.6% (2/7) with both techniques. Finally, the presence of MAP was identified in 42.9% (3/7) of semen samples with nPCR; however, it was not detected through culture. In conclusion, the presence of MAP was identified in lymphatic, digestive tissue, and semen; the presence of MAP was reported for the first time in epididymis, Cowper gland, prostate and testicles of infected Pelibuey rams.

Caprine arthritis encephalitis: an example of risk assessment for embryo trading.

The risk of transmission of caprine arthritis encephalitis virus (CAEV) during embryo transfer has been demonstrated in vivo through the detection of CAEV proviral DNA in: (1) flushing media for embryo collection; (2) cells of the cumulus oophorus surrounding the oocytes, ovarian follicle, oviduct and uterine tissues; and (3) testis, epididymis, vas deferens and vesicular glands. Experimentally infected embryos without a zona pellucida (ZP), washed 10 times with Minimum Essential Media (MEM) and 5% Fetal Calf Serum (FCS) solution, were capable of transmitting CAEV. In vitro we demonstrated that granulosa, oviductal, epididymal and embryo cells are fully susceptible to CAEV infection and allow active replication. However, AI with in vitro-infected semen can result in the production, after ten washing, of CAEV-free embryos, and ten washing in vitro- or in vivo-infected embryos with an intact ZP, or ten washing oocytes with an intact ZP, resulted in the production of virus-free female gametes or embryos that can be used for IVF or embryo transfer. Therefore, we have demonstrated that: (1) that CAEV-free embryos can be produced by IVF using spermatozoa infected in vitro by CAEV; and (2) embryo transfer can be used under field conditions to produce CAEV-free kids from CAEV-infected biological mothers.