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<jats:title>Abstract</jats:title><jats:p>In this study, the relationships between post‐thaw bull sperm characteristics and hyperketonemic conditions after coincubation with cow plasma or media were determined to investigate if such a condition could affect bull sperm characteristics. Two experiments were conducted. In experiment 1, blood samples were collected from 31 cows to prepare plasma. Cows were independently categorized into two groups according to plasma β‐hydroxybutyrate (BHB) concentrations (above or below 1.2 mM). Thawed bull semen was diluted and incubated with diluted plasma; motility parameters were evaluated using Computer Assisted Semen Analysis (CASA). In experiment 2, a pooled sample of thawed semen was diluted and divided into three aliquots: without BHB (control) and treated with either 1.2 mM (1.2) or 3 mM (3) BHB. In addition to motility, flow cytometric analyses were carried out. In experiment 1, the overall motility decreased significantly in plasma containing high (≥1.2 mM) BHB compared to plasma containing low (&lt;1.2 mM) BHB. In experiment 2, the overall motility tended to be lower in BHB (3 mM)‐supplemented samples. The supplementation of 3 mM BHB increased the proportion of live superoxide‐positive sperm and sperm with high mitochondrial potential, while the DNA fragmentation index decreased.</jats:p>

<jats:title>Abstract</jats:title><jats:p>This study aimed to compare the postpartum uterine dynamics of primiparous precocious (PP), primiparous conventional (PC) and multiparous conventional (MC) <jats:italic>Bos indicus</jats:italic> beef cows. For this purpose, PP (<jats:italic>n</jats:italic> = 8), PC (<jats:italic>n</jats:italic> = 18) and MC (<jats:italic>n</jats:italic> = 12) cows were enrolled in this study. These cows were evaluated at 20 and 10 days prepartum and weekly from parturition to 42 days postpartum (DPP). During this period, body weight (BW), subcutaneous fat thickness (SFT) and serum concentrations of glucose, β‐hydroxybutyrate, albumin and haptoglobin were measured. Proportion of polymorphonuclear (PMN) cells, and abundance of mRNA transcripts of genes involved in uterine inflammation and uterine health were evaluated. The PP cows had lower (<jats:italic>p</jats:italic> &lt; .05) BW and SFT than that for PC and MC cows during the study period. The serum concentration of albumin after 35 DPP was lower (<jats:italic>p</jats:italic> &lt; .05) in PP cows. The PP cows had the highest proportion of PMN on 28 and 35 DPP compared to PC and MC cows. The relative mRNA abundance of IL‐1β and IL‐8 increased after 21 DPP in PP cows compared to the other groups. The PC had the highest, MC had an intermediate, and PP cows had the lowest relative abundance of IL10 mRNA. Overall, these findings indicated that uterine inflammation was more pronounced in PP cows. Moreover, based on the proportion of PMN and abundance of transcripts associated with inflammation in the uterus, PP cows may require a longer period to recover their uterine health after calving.</jats:p>

<jats:title>Abstract</jats:title><jats:p>A ten‐year‐old mixed breed bitch was presented for a tissue prolapse protruding from her vulva. Following detailed examination and stabilization, the ovaries and uterine horns were removed by laparotomy, whereas the prolapsed tissue identified as uterus including cervix was removed vaginally. Histology confirmed uterine prolapse, a rare condition in bitches usually found shortly after birth especially due to dystocia. In contrast, the present case was found in a nulliparous non‐pregnant bitch. Diagnostic and therapeutic approaches, including microbiological and histological findings, are described and discussed critically.</jats:p>

<jats:title>Abstract</jats:title><jats:p>Anti‐Müllerian hormone (AMH) is secreted by granulosa cells of healthy, growing follicles and is positively correlated with the ovarian reserve. Maternal and environmental factors, such as nutrition, disease, parity and endocrine disruptors, are thought to have a profound impact on ovarian reserve development during early foetal life. For genetic progress, it can be advantageous to breed dairy replacements from heifers to expedite the generation interval; however, there is some evidence that nulliparous animals produce female offspring with smaller ovarian reserves compared with multiparous animals. The objective of this prospective, observational study was to determine whether maternal growth in the pre‐conception and early gestational period of nulliparous dairy heifers is associated with pre‐weaning AMH concentrations in their female offspring. Our hypothesis was that excessive growth in this period would negatively impact AMH concentrations. Seasonal, pasture‐based dairy heifer calves (<jats:italic>n</jats:italic> = 156) born from nulliparous dams, from six Irish farms, were blood sampled at an average of 60 days of age in spring 2022 and tested for AMH. Mixed‐effects linear regression models were constructed with Box‐Cox transformed AMH concentration as the dependent variable. The independent variables tested included maternal average daily gain (ADG) from pre‐breeding examination (PBE) to pregnancy diagnosis (PD) between 30 and 60 days in calf (DIC), ADG from PBE to PD over 60 DIC and ADG between the two PDs. Calf breed and age at sampling were forced into the models, and the farm was treated as a random effect in all models. We found that as ADG increased from the pre‐breeding period to their first PD visit, the AMH concentration in their offspring reduced. However, ADG explained only a small amount of the variation in AMH concentrations (marginal <jats:italic>R</jats:italic><jats:sup>2</jats:sup> = 0.041). In conclusion, the results of our study suggest that excessive growth prior to conception and in early gestation of nulliparous heifers could impact the ovarian reserve of their female offspring, and may imply that farmers should avoid excessive growth in the immediate pre‐breeding and early gestational periods.</jats:p>

<jats:title>Abstract</jats:title><jats:p>This study analysed data from a commercial swine herd in Thailand equipped with a free‐farrowing housing system, comprising 17,196 piglets from 1318 litters, to explore the impact of sow and litter characteristics on the piglet birth weight and the incidence of stillbirth. The piglets were classified into four groups based on the total number of piglets born per litter (TB): ≤9 (<jats:italic>n</jats:italic> = 1434), 10–12 (<jats:italic>n</jats:italic> = 3232), 13–15 (<jats:italic>n</jats:italic> = 6537) and ≥16 (<jats:italic>n</jats:italic> = 5993). Sows were classified into four groups based on parity number: 1, 2–4, 5–7 and ≥8. The piglets were categorized into quartiles based on their birth order ranking: Q1 (<jats:italic>n</jats:italic> = 4786), Q2 (<jats:italic>n</jats:italic> = 4143), Q3 (<jats:italic>n</jats:italic> = 3808) and Q4 (<jats:italic>n</jats:italic> = 4456). Piglet birth weight was individually measured before colostrum ingestion. On average, TB, the number of live‐born piglets and the incidence of stillbirth were 13.1 ± 3.7, 11.5 ± 3.8 and 6.3%, respectively. Among these litters, 26.6% had TB numbers ≥16. The average piglet birth weight was 1.37 ± 0.36 kg, with 18.3% of piglets weighing ≤1.0 kg at birth. Piglet birth weight was influenced by birth order ranking, as Q4 piglets were found to be heavier than piglets born in Q1–Q3 (<jats:italic>p</jats:italic> &lt; .001). Moreover, the percentage of piglets with a birth weight of ≤1.0 kg increased from 5.9% in litters with TB of ≤9–25.3% in litters with TB of ≥16 (<jats:italic>p</jats:italic> &lt; .001). Additionally, primiparous sows had lower piglet birth weights compared to sows with parity numbers 2–4, 5–7 and ≥8 (<jats:italic>p</jats:italic> &lt; .001). Piglets born in the fourth quartile (Q4) had a higher risk of stillbirth compared to those born in the first (Q1), second (Q2) and third (Q3) quartiles (12.5% vs. 2.2%, 4.1% and 6.6%, respectively, <jats:italic>p</jats:italic> &lt; .001). The incidence of stillbirth in litters with TB ≥16 was also higher than that in litters with TB ≤9 and 10–12 (<jats:italic>p</jats:italic> &lt; .05). Furthermore, sows with parity numbers ≥8 had a higher incidence of stillbirth (9.7%) compared to primiparous sows (4.0%, <jats:italic>p</jats:italic> &lt; .001), sows with 2–4 parity (5.2%, <jats:italic>p</jats:italic> &lt; .001) and sows with 5–7 parity (7.6%, <jats:italic>p</jats:italic> = .003). In summary, a high incidence of stillbirth was found in piglets born in the last quartile of litters, in litters with &gt;16 piglets and for sows with parity numbers ≥8. Piglets born in the last quartile of litters were heavier than those born in the first to third quartiles.</jats:p>

<jats:title>Abstract</jats:title><jats:p>The aim of this study was to investigate the relationship between the levels of insulin‐like growth factor‐1 (IGF‐1) in serum and seminal plasma and the characteristics of semen in Beetal bucks (<jats:italic>Capra hircus</jats:italic>). A total of 12 adult Beetal bucks were involved in the study, with each buck providing six ejaculates collected using a standard artificial vagina (<jats:italic>n</jats:italic> = 72 total). Only qualified semen samples (volume of 0.7 mL, a mass motility rating of 3+ or higher on a 0–+ scale, and individual progressive motility of 80% or more) divided into three fractions were processed for estimation of IGF‐1 and other seminal parameters like motility, viability, acrosome integrity, sperm abnormality and superoxide dismutase (SOD) activity. The first and second fraction were diluted and extended with Optixcell extender (1:15 ratio). The first ejaculate fraction was processed for studying fresh semen parameters and the second fraction was cryopreserved for evaluating frozen semen parameters. French mini straws (0.25 mL) were used for semen filling, and polyvinyl alcohol powder of different colours was used for sealing the extended semen. The third fraction of each ejaculate was centrifuged at room temperature (1100 × g for 7 min) to separate the seminal plasma. Additionally, blood samples were taken from each buck on the same day as semen collection, resulting in a total of 36 blood samples. The results revealed a significant positive correlation (<jats:italic>r</jats:italic> = .4243; <jats:italic>p</jats:italic> &lt; .05) between the concentration of IGF‐1 in both serum and seminal plasma of the Beetal bucks. Furthermore, the concentration of IGF‐1 in serum showed significant positive correlations with sperm viability (<jats:italic>r</jats:italic> = .554; <jats:italic>p</jats:italic> &lt; .05), acrosome integrity (<jats:italic>r</jats:italic> = .527; <jats:italic>p</jats:italic> &lt; .05), post‐thaw sperm motility (<jats:italic>r</jats:italic> = .407; <jats:italic>p</jats:italic> &lt; .01), post‐thaw sperm viability (<jats:italic>r</jats:italic> = .426; <jats:italic>p</jats:italic> &lt; .01) and post‐thaw acrosome integrity (<jats:italic>r</jats:italic> = .333; <jats:italic>p</jats:italic> &lt; .05). However, it had a significant negative correlation with SOD activity in fresh semen (<jats:italic>r</jats:italic> = −0.458; <jats:italic>p</jats:italic> &lt; .01). Moreover, the concentration of IGF‐1 in seminal plasma demonstrated significant positive correlations with individual progressive motility (<jats:italic>r</jats:italic> = .341; <jats:italic>p</jats:italic> &lt; .05), sperm viability (<jats:italic>r</jats:italic> = .527; <jats:italic>p</jats:italic> &lt; .05), acrosome integrity (<jats:italic>r</jats:italic> = .539; <jats:italic>p</jats:italic> &lt; .05), sperm plasma membrane integrity (<jats:italic>r</jats:italic> = .464; <jats:italic>p</jats:italic> &lt; .05), post‐thaw sperm motility (<jats:italic>r</jats:italic> = .644; <jats:italic>p</jats:italic> &lt; .01), post‐thaw sperm viability (<jats:italic>r</jats:italic> = .643; <jats:italic>p</jats:italic> &lt; .01), post‐thaw acrosome integrity (<jats:italic>r</jats:italic> = .487; <jats:italic>p</jats:italic> &lt; .01) and post‐thaw sperm plasma membrane integrity (<jats:italic>r</jats:italic> = .521; <jats:italic>p</jats:italic> &lt; .01). Additionally, it showed a significant negative correlation with SOD activity in both fresh semen (<jats:italic>r</jats:italic> = −0.714; <jats:italic>p</jats:italic> &lt; .01) and frozen semen (<jats:italic>r</jats:italic> = −0.558; <jats:italic>p</jats:italic> &lt; .01) of Beetal bucks. Based on these findings, IGF‐1 in seminal plasma can be considered as a potential biomarker for the selection of bucks for breeding purposes.</jats:p>

<jats:title>Abstract</jats:title><jats:p>Milk production traits as the most important economic traits of dairy cows, they directly reflect the benefits of breeding and the economic benefits of pasture. In this study, A disintegrin and metalloproteinase‐12 (<jats:italic>ADAM12</jats:italic>), Parkinson's disease gene 2 (<jats:italic>PRKN</jats:italic>) and dipeptidyl peptidase‐like protein subtype 6 (<jats:italic>DPP6</jats:italic>) polymorphism in 384 Chinese Holstein cows were detected by time‐of‐flight mass spectrometry and through statistical analysis using software such as Popgene 32, SAS 9.4 and Origin 2022, the relationship between single nucleotide polymorphisms (SNPs) of three genes with four milk production traits such as daily milk yield (DMY), milk fat percentage (MFP), milk protein percentage (MPP) and somatic cell score (SCS) was verified at molecular level. The results showed that four polymorphic loci (116,467,133, 116,604,487, 116,618,268 and 116,835,111) of <jats:italic>DPP6</jats:italic> gene, two polymorphic loci (97,665,052 and 97,159,837) of <jats:italic>PRKN</jats:italic> gene and two polymorphic loci (45,542,714 and 45,553,888) of <jats:italic>ADAM12</jats:italic> gene were detected. <jats:italic>PRKN</jats:italic>‐97665052, <jats:italic>DPP6</jats:italic>‐116467133, <jats:italic>ADAM12</jats:italic>‐45553888, <jats:italic>DPP6</jats:italic>‐116604487 and <jats:italic>DPP6</jats:italic>‐116835111 were all in Hardy–Weinberg equilibrium state (<jats:italic>p</jats:italic> &gt; .05). <jats:italic>ADAM12</jats:italic>‐45542714, <jats:italic>PRKN‐</jats:italic>97159837 and <jats:italic>PRKN‐</jats:italic>97665052 were moderately polymorphic (0.25 ≤ <jats:italic>PIC</jats:italic> &lt;0.50) in Holstein. It is evident that the selection potential and genetic variation of these five loci are relatively large, and the genetic richness is relatively high. The correlation analysis of different genotypes between these eight loci and milk production traits of Holstein showed that <jats:italic>ADAM12</jats:italic>‐45542714 and <jats:italic>DPP6</jats:italic>‐116835111 (<jats:italic>p</jats:italic> &lt; .01) had an extremely significant effects on the DMY of Chinese Holstein in Ningxia, while <jats:italic>PRKN</jats:italic>‐97665052 had an extremely significant effect on MFP (<jats:italic>p</jats:italic> &lt; .01). The effect of <jats:italic>PRKN</jats:italic>‐97665052 and <jats:italic>DPP6</jats:italic>‐116467133 on MPP of Holstein were extremely significant (<jats:italic>p</jats:italic> &lt; .01). <jats:italic>DPP6‐</jats:italic>116618268 had an extremely significant effect on the SCS of Holstein in Ningxia (<jats:italic>p</jats:italic> &lt; .01), and <jats:italic>AA</jats:italic> genotype individuals showed a higher SCS than <jats:italic>GG</jats:italic> genotype individuals; the other two loci (<jats:italic>ADAM12</jats:italic>‐45553888 and <jats:italic>DPP6‐</jats:italic>116604487) had no significant effects on milk production traits of Holstein (<jats:italic>p</jats:italic> &gt; .05). In addition, through the joint analysis of <jats:italic>DPP6</jats:italic>, <jats:italic>PRKN</jats:italic> and <jats:italic>ADAM12</jats:italic> gene loci, it was found that the interaction effect between the three gene loci could significantly affect the DMY, SCS (<jats:italic>p</jats:italic> &lt; .01) and MPP (<jats:italic>p</jats:italic> &lt; .05). In conclusion, several different loci of <jats:italic>DPP6</jats:italic>, <jats:italic>PRKN</jats:italic> and <jats:italic>ADAM12</jats:italic> genes can affect the milk production traits of Holstein to different degrees. <jats:italic>PRKN</jats:italic>, <jats:italic>DPP6</jats:italic> and <jats:italic>ADAM12</jats:italic> genes can be used as potential candidate genes for milk production traits of Holstein for marker‐assisted selection, providing theoretical basis for breeding of Holstein.</jats:p>

<jats:title>Abstract</jats:title><jats:p>Follistatin (FST), a member of the transforming growth factor‐β (TGF‐β) superfamily, has been identified as an inhibitor of follicle‐stimulating hormone. Previous studies showed that it plays an important role in animal reproduction. Therefore, this study aims to investigate its effect on the maturation of buffalo oocytes in vitro, and the underlying mechanism of FST affecting oocyte maturation was also explored in buffalo cumulus cells. Results showed that FST was enriched in the ovary and expressed at different stages of buffalo ovarian follicles as well as during oocyte maturation and early embryo development. The FST expression level was up‐regulated in MII buffalo oocytes compared with the GV stage (<jats:italic>p</jats:italic> &lt; .05). To study the effects of FST on buffalo oocytes' maturation and early embryonic development, we added the pcD3.1 skeleton vector and PCD3.1‐EGFP‐FST vector into the maturation fluid of buffalo oocytes, respectively. It was demonstrated that FST promoted the in vitro maturation rate of buffalo oocytes and the blastocyst rate of embryos cultured in vitro (<jats:italic>p</jats:italic> &lt; .05). By interfering with FST expression, we discovered that FST in cumulus cells plays a crucial role in oocyte maturation. Interference with the FST expression during the buffalo oocyte maturation did not affect the first polar body rate of buffalo oocyte (<jats:italic>p</jats:italic> &gt; .05). In contrast, the location of mitochondria in oocytes was abnormal, and the cumulus expansion area was reduced (<jats:italic>p</jats:italic> &lt; .05). After parthenogenetic activation, the cleavage and blastocyst rates of the FST‐interfered group were reduced (<jats:italic>p</jats:italic> &lt; .05). Furthermore, RT‐qPCR was performed to investigate further the underlying mechanism by which FST enhances oocyte maturation. We found that overexpression of FST could up‐regulate the expression level of apoptosis suppressor gene Bcl‐2 and TGF‐β/SMAD pathway‐related genes TGF‐β, SMAD2, and SMAD3 (<jats:italic>p</jats:italic> &lt; .05). In contrast, the expression levels of SMAD4 and pro‐apoptotic gene BAX were significantly decreased (<jats:italic>p</jats:italic> &lt; .05). The FST gene could affect buffalo oocyte maturation by regulating the oocyte mitochondria integrity, the cumulus expansion, cumulus cell apoptosis, and the expression levels of TGF‐β/SMAD pathway‐related genes.</jats:p>

<jats:title>Abstract</jats:title><jats:p>This study evaluated the effect of supplementing IVM media with γ‐oryzanol (ORY), a nutraceutical derived from rice bran oil, on the development of bovine oocytes and hindering the compromising effect of redox imbalance. An in vitro model of the bovine cumulus‐oocyte complex was used for the evaluation of nuclear maturation and development. Antioxidant activity was investigated by assessing the level of ROS (Reactive Oxygen Species) and GSH (glutathione) in oocytes and quantitative changes in gene expression in matured oocytes and their respective cumulus cells. ORY supplementation increased the proportion of MII oocytes, cleaved embryos, and total blastocysts (<jats:italic>p</jats:italic> &lt; .05) and was linked to higher and lower levels of intracellular GSH and ROS, respectively (<jats:italic>p</jats:italic> &lt; .05). The treated oocytes and their respective cumulus‐granulosa cells showed a modulation in the expression of genes related to apoptosis (downregulation of <jats:italic>BAX</jats:italic> and <jats:italic>CHOP</jats:italic>) and oxidative stress (upregulation of <jats:italic>NRF2</jats:italic>, <jats:italic>CAT</jats:italic>, and <jats:italic>SOD</jats:italic>). Also, relative upregulation of <jats:italic>OCT‐4</jats:italic> and <jats:italic>IGF2R</jats:italic> in treated oocytes was concomitant with higher subsequent development in terms of cleavage and total blastocyst rates (<jats:italic>p</jats:italic> &lt; .05). Based on our findings, it appears that ORY supplementation can improve the nuclear maturation and development of bovine oocytes into blastocysts and augment their enzymatic and non‐enzymatic antioxidant systems, maintaining the Redox balance and high enzymatic activity against ROS generation.</jats:p>

<jats:title>Abstract</jats:title><jats:p>This study evaluated the effects of melatonin (MLT) and L‐carnitine supplementation on sperm quality and antioxidant capacity during chilled and cryopreservation. Twenty‐four ejaculates were collected from six <jats:italic>Damascus</jats:italic> bucks, 4 ejaculates each, from mid‐September to mid‐October 2022. The pooled semen from each collecting session was divided into 5 equal aliquots after being diluted (1:10) with Tris‐citric acid egg yolk extender. The first aliquot served as a control (treatment‐free). MLT was added to the second and third aliquots at low and high doses (LD: 4 and HD: 8 μL/mL) (v/v), respectively, while L‐carnitine (LC) was added to the fourth and fifth aliquots at the same aforementioned doses. The aliquots were stored at 4°C for 48 h to assess sperm physical and morphological characteristics, alongside lipids peroxidase (LP) production and glutathione peroxidase (GPX) activity. The optimum doses of MLT and LC that showed potential for maintaining sperm characteristics throughout the chilled storage period were further investigated for protecting the spermatozoa after exposure to cryopreservation stress compared to the control. The results showed higher sperm motility (%) in the MLT‐HD group, higher (<jats:italic>p</jats:italic> ≤ .05) sperm viability (%) in the MLT‐LD, and both aliquots of LC at T<jats:sub>24</jats:sub> hours of chilled preservation. Normal sperm (%) was higher (<jats:italic>p</jats:italic> ≤ .05) in both LC‐LD and MLT‐LD groups than other groups, while sperm acrosome integrity (%) was higher (<jats:italic>p</jats:italic> ≤ .05) in the LC‐LD group. Morphological abnormalities (%) were improved (<jats:italic>p</jats:italic> ≤ .05) in all treated aliquots compared with control. The mean value of GPX activity was higher (<jats:italic>p</jats:italic> ≤ .05) in both MLT groups, while the concentration of LP increased (<jats:italic>p</jats:italic> ≤ .05) in the LC‐HD or control groups. Furthermore, supplementing buck sperm medium with 4 μL/mL of MLT or LC improved (<jats:italic>p</jats:italic> &lt; .05) the sperm characteristics and decreased (<jats:italic>p</jats:italic> &lt; .05) DNA fragmentation index after thawing.</jats:p>