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Digital, The effect of L-165041 (PPARδ-agonist) on decreasing apoptosis and intracellular lipid content was assessed in fresh and vitrified–warmed in vitro-produced bovine embryos. It was hypothesised that the addition of L-165041 to the culture medium enhances development and cryopreservation. Oocytes were allocated to one of two treatments: control-standard culture medium, or L-165041 added to the medium on day 1 with no media change. Ultrastructure, cleavage, and blastocyst rates were evaluated in fresh, and in post-vitrification cultured embryos by optical and electronic microscopy. A subset of fresh embryos were fixed for TUNEL assay and for Sudan-Black-B histochemical staining. Vitrified–warmed embryos were assessed using MALDI-MS technique. Cleavage and blastocyst rates (control 49.4 ± 5.2, L-165041 51.8 ± 4.3) were not influenced by L-165041. The proportion of inner cell mass cells (ICM) was higher in fresh embryos, and the rate of total and ICM apoptosis was lower in L-165041. In warmed-embryos, total and ICM apoptosis was lower in L-165041. The overall hatching rate was higher in L-165041 (66.62 ± 2.83% vs 53.19 ± 2.90%). There was less lipid accumulation in fresh L-165041-embryos. In conclusion, the use of L-165041 is recommended to improve the viability of in vitro-derived bovine embryos., Ciencias Agropecuarias

SummaryThis study aimed to evaluate the relationship between antral follicular count (AFC) and ovarian volume (OV), preantral follicular population and survival, meiotic progression and ultrastructure of cumulus–oocyte complexes (COCs) after in vitro maturation. In experiment 1, the relationship between AFC and preantral follicle population and survival was evaluated by classical histology. In experiment 2, the relationship among AFC, OV, ability of oocytes to resume meiosis and ultrastructure of in vitro matured bovine COCs was studied. A positive correlation (P < 0.05) between AFC and the numbers of healthy primordial, degenerate and total follicles was observed, as well as with healthy secondary follicles and total follicles. The numbers of grades I and II oocytes in ovaries of high AFC class were higher compared with those with intermediate or lower AFC. After in vitro maturation, COCs from ovaries of high AFC had a higher percentage of oocytes in metaphase II compared with those of intermediate and low AFC (P < 0.0001). Ovaries of intermediate AFC had a higher percentage of oocytes in metaphase II compared with ovaries with low AFC (P < 0.0001). The proportion of oocytes in metaphase I, telophase I and anaphase I in COCs from ovaries of intermediate AFC (26.04%) was higher (P < 0.05) compared with that seen in COCs of ovaries with high (8.55%) and low (14.15%) AFC. No differences in the ultrastructure of oocytes were seen. In conclusion, after in vitro maturation, cow ovaries with high AFC have higher numbers of oocytes that reach in metaphase II (MII), but they also have higher numbers of degenerated primordial and primary follicles.

The effect of L-165041 (PPARδ-agonist) on decreasing apoptosis and intracellular lipid content was assessed in fresh and vitrified-warmed in vitro -produced bovine embryos. It was hypothesised that the addition of L-165041 to the culture medium enhances development and cryopreservation. Oocytes were allocated to one of two treatments: control-standard culture medium, or L-165041 added to the medium on day1 with no media change. Ultrastructure, cleavage, and blastocyst rates were evaluated in fresh, and in post-vitrification cultured embryos by optical and electronic microscopy. A subset of fresh embryos were fixed for TUNEL assay and for Sudan-Black-B histochemical staining. Vitrified-warmed embryos were assessed using MALDI-MS technique. Cleavage and blastocyst rates (control 49.4±5.2, L-165041 51.8±4.3) were not influenced by L-165041. The proportion of inner cell mass cells (ICM) was higher in fresh embryos, and the rate of total and ICM apoptosis was lower in L-165041. In warmed-embryos, total and ICM apoptosis was lower in L-165041. The overall hatching rate was higher in L-165041 (66.62±2.83% vs 53.19±2.90%). There was less lipid accumulation in fresh L-165041-embryos. In conclusion, the use of L-165041 is recommended to improve the viability of in vitro -derived bovine embryos.

This study aimed to evaluate the relationship between antral follicular count (AFC) and ovarian volume (OV), preantral follicular population and survival, meiotic progression and ultrastructure of cumulus-oocyte complexes (COCs) after in vitro maturation. In experiment 1, the relationship between AFC and preantral follicle population and survival was evaluated by classical histology. In experiment 2, the relationship among AFC, OV, ability of oocytes to resume meiosis and ultrastructure of in vitro matured bovine COCs was studied. A positive correlation (P0.05) between AFC and the numbers of healthy primordial, degenerate and total follicles was observed, as well as with healthy secondary follicles and total follicles. The numbers of grades I and II oocytes in ovaries of high AFC class were higher compared with those with intermediate or lower AFC. After in vitro maturation, COCs from ovaries of high AFC had a higher percentage of oocytes in metaphase II compared with those of intermediate and low AFC (P0.0001). Ovaries of intermediate AFC had a higher percentage of oocytes in metaphase II compared with ovaries with low AFC (P0.0001). The proportion of oocytes in metaphase I, telophase I and anaphase I in COCs from ovaries of intermediate AFC (26.04%) was higher (P0.05) compared with that seen in COCs of ovaries with high (8.55%) and low (14.15%) AFC. No differences in the ultrastructure of oocytes were seen. In conclusion, after in vitro maturation, cow ovaries with high AFC have higher numbers of oocytes that reach in metaphase II (MII), but they also have higher numbers of degenerated primordial and primary follicles.

The intense selection of characteristics related to animal performance may affect the reproductive efficiency of beef cows negatively. Selection for fertility along with production traits is, therefore, readily justifiable. The objective of the present study was to evaluate possible relationships among AFC, serum AMH concentrations and external morphometrics with reproductive efficiency in Tabapua (a Brazilian Bos indicus beef breed) females. Antral follicle counts and external morphometrics were measured in nulliparous (n = 162), primiparous (n = 80) and multiparous (n = 351) cows, from four farms. Age at first parturition (AFP), parturition interval (PI), maternal ability (MA), precocity, rusticity and survival (PRS) and overall reproductive efficiency (RE) were evaluated according to the Brazilian Association of Zebu Breeders (ABCZ) data bank indexes. A single blood sample per animal was collected at random stages of the reproductive cycle from a subset of animals (nulliparous = 84, primiparous and multiparous = 136) to determine serum AMH concentrations. The AFC classes were defined as lesser ( 50). Correlations between all variables and the effects of parity and AFC on reproductive traits, AMH and external genitalia size were analyzed by the PROCORR and by the PROCGENMOD procedures of SAS® (SAS, Cary, NC, USA), respectively. Antral follicle count did not differ (P = 0.71) among nulliparous (38.6 ± 23.96) primiparous (47.54 ± 26.16) and multiparous (41.08 ± 25) cows and was negatively correlated with pregnancy interval (PI), such that, as PI decreased (r = −0.28; P

The objective was to assess the influence of polyunsaturated fatty acid supplementation on the quality of fresh, cooled, and frozen-thawed stallion semen. Ten stallions received their normal diet (control group) or normal diet plus 150 mL of polyunsaturated fatty acid (PUFA) linseed-based oil (PUFA group). Semen was collected every 15 days during 60 days. Stallions were reversed across the treatments after a sixty-day interval. Semen was evaluated at 2, 6, 12, and 24 hours after cooling and 24 hours after freezing. Motility (MOT), vigor, membrane viability, morphology, acrosome integrity, and osmotic tolerance test (OTT) were evaluated. In the frozen-thawed semen, sperm dynamic characteristics were analyzed by computer-assisted sperm analysis and thiobarbituric acid reactive substances (TBARs) determined. The effects of treatment, time, semen type, and their interactions were submitted to PROCMIX (SAS) and means compared by the Tukey test. There was no treatment effect on the quality of fresh and cooled semen. However, frozen-thawed semen MOT, vigor, and OTT were superior ( P P = .002) in the PUFA group after 15, 30, 45, and 60 days from the beginning of supplementation. Thus, sperm may become more susceptible to the reactive oxygen species, probably due to the incorporation of polyunsaturated fat in the cell membrane. The addition of PUFA-enriched oil may be an alternative for improving frozen-thawed semen quality by increasing its MOT and resistance to osmotic changes to which sperm cells are submitted during the freezing process.

It was hypothesized that Brahman cows have greater in vitro embryo production than Simmental cows, and that regardless of breed, antral follicle count (AFC) and vulvar width (VW) are positively associated with oocyte quality, embryo number and pregnancy rate. Brahman (Bos taurus indicus, n = 184) and Simmental (Bos taurus taurus, n = 99) cows were classified as having a large AFC (Brahman: ≥50, Simmental: ≥25); intermediate AFC (Brahman:30 and ≤49; Simmental:16 and24) and small AFC (Brahman:30, Simmental: ≤15). The VW was considered to be large (Brahman:123 mm, Simmental:80 mm); intermediate (Brahman:102 mm and ≤123 mm; Simmental:65 mm and ≤80 mm) and small (Brahman: ≤102 mm and Simmental: ≤65 mm). For each group of cows, ovum pick up, oocyte in vitro maturation, fertilization and embryo culture were performed. Embryo transfers were subsequently performed and pregnancy rate was evaluated. Brahman cows had a larger number of viable oocytes, greater efficiency of embryo production and greater pregnancy rate (P0.05) than Simmental cows. Pregnancy percentages in Simmental cows with a small AFC were greater than those cows with an intermediate and large AFC. With the Brahman cows, there was no difference in pregnancy rate among the cows with different AFC classifications. In contrast to Simmental cows, Brahman cows with an intermediate VW had a greater pregnancy percentage than those with a small and large AFC. In conclusion, Brahman cows have a larger AFC, number of viable oocytes, and pregnancy rate than Simmental cows.

This study evaluated the presence of polyunsaturated fatty acids in circulating blood and in the ovarian follicular fluid of mares, after supplementation of the diet with linseed oil. Six Mangalarga Marchador mares, weighing 397.00±31.89 kg, were kept on native pasture, and assigned to the current study. In a switch over design, mares were randomly allocated to receive 150 ml of vegetable oil daily, containing polyunsaturated fatty acids n3 (62.23 g ALA, 20.34 g LA, 2.27 g EPA, 2.32 g DHA), (n=3) or no supplementation (n=3) in two replicates. Blood and follicular fluid samples were taken on the first day (D0) and every 30 days until the end of the supplementation period (D60). After 60 days of supplementation, mares were switched across the treatments. Plasma concentrations of linolenic acid in total fatty acids were higher (P=0.006) in the supplemented compared to the control group (1.89±0.13 vs. 1.49±0.13%). There were positive correlations between plasma linoleic acid and follicular fluid arachidonic acid (P=0.0106; r2=0.13) and between plasma alpha linolenic acid and follicular fluid EPA (P=0.0004; r2=0.2544). Data indicated a low to moderate relationship between the dietary linseed-based oil supplementation studied and circulating and follicular fluid polyunsaturated fatty acids contents in mares.

An innovative technology that currently tries to gain market share are milking robots. Despite the high costs, robotic milking may produce benefits for the farmer and for animals submitted to this system. The objective of this study was to perform a literature review on the use of robotic milking of dairy cows, addressing aspects such as implementation and functioning of the system and effect on milking frequency, milk production and composition, somatic cell count, mastitis, reproduction, and animal welfare. The results showed that the programming of the robot depends on the number of animals to be milked daily and on the milking frequency adopted in each batch. The implementation of the system mainly depends on the site where the facilities will be built and already existing structures on the farm. The milking frequency is higher for high-producing cows and at the beginning of lactation and is influenced by the palatability of the concentrate offered at the time of milking. Frequencies higher than three times per day reduce total milk fat production and increase the concentration of free fatty acids. The incidence of mastitis and somatic cell count tend to increase in the first three months after implementation of the system. After the second year, udder health tends to improve, normalizing mastitis incidence and somatic cell count. Reproduction of the cows is not affected if dry matter intake compensates the higher energy expenditure required for higher milk productions. Robotic milking improves cow welfare since the animals voluntarily turn to the robot when they feel discomfort.

Made available in DSpace on 2022-04-28T19:05:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-01-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) The aim of this study was to investigate possible relationships between morphometric characteristics of the bovine reproductive tract and measures of fertility, such as antral follicle counts (AFCs) and the number and quality of recovered cumulus oophorus complexes (COCs). First, the genital tracts of 360 abattoir Zebu/Holstein crossbred cows were studied. Rima vulvae length (RL), vulvar width (VW), ovarian size, pelvic fat, number of aspirated COCs, and AFCs were recorded. An index of COC quality (CQI) was established, which weighed overall gamete quality based on the IETS classification. Second, the same external measurements and AFCs (by ultrasound) were analyzed in live Tabapuã (Zebu) cows (n = 48). Relative RL and vulvar width were defined as the original measurements divided by the body weight of each cow for statistical analyses. In abattoir tracts, the AFC was smaller (P < 0.05) in animals with large VW (34.88 ± 3.50) than in animals with small (45.71 ± 2.57) and intermediate (42.25 ± 2.45) VW. The CQI was higher (P < 0.05) when the pelvic fat score was intermediate (3.22 ± 0.06) than that in lean (2.99 ± 0.08) and obese (2.90 ± 0.12) animals. The CQI was higher (P < 0.05) in the intermediate ovary group (3.19 ± 0.08) than in the small (2.96 ± 0.08) and large (2.95 ± 0.09) ovary groups. In live cows, the AFC was higher (P = 0.035) in females with large relative rima length than in those with small relative rima length (18.96 ± 1.97 and 14.76 ± 1.51, respectively). Similarly, AFC was greater (P = 0.0001) in females with large relative VW than in those with small relative VW (20.08 ± 1.36 and 11.16 ± 1.60, respectively). In conclusion, larger external genitalia relative to body size were good predictors of the ovarian follicular reserve in live animals. Division of Agriculture Polytechnic Institute of Gaza Polytechnic Campus, District of Chókwè Department of Animal Science Federal University of Lavras (UFLA) Department of Animal Reproduction and Veterinary Radiology University of São Paulo State In Vitro Brazil

The aims of this study were to decrease the apoptotic index and increase cryotolerance of bovine embryos produced in vitro with the addition of 1 µM docosahexaenoic acid (DHA). On Day 1, presumed zygotes were cultivated with 1µM DHA (Sigma, St. Louis, MO, USA; n=437), and without the agonist (control group; n=450). The cleavage rate (%) was evaluated on Day 2 and the development of blastocysts on Day 7. Embryos before and after vitrification were fixed for the TUNEL trial. After vitrification, the embryos were heated and re-cultivated to evaluate the hatching rate at 12, 24, 36, 48, 60, and 72h. A sample of re-cultivated embryos at 12h of DHA (n=5), and without the agonist (control group; n=6), was frozen for mass spectrometry (MALDI-MS). Statistical analyses of deviance were carried out considering generalized mixed linear models, and the effect of the collection day (block) was considered as random. For the count variables, the Poisson distribution and the log link function were considered. In the cases of variables represented by rates, binomial distribution and the logit link function were used. In the study of cryotolerance, ANOVA of the hatching rate for each one of the times evaluated was carried out. In cases of significance of the effect of treatments, the Dunnett test was applied to compare treatments. Multivariate and univariate statistical models were used for analysis of MALDI-MS. All analyses were made using the GLIMMIX procedure of the SAS software (SAS Institute Inc., Cary, NC, USA). The cleavage rate was not different between the groups (P>0.05) and the production of blastocysts was lower in the DHA group (P0.05) compared with the control group. Phosphatidylcholine [phosphatidylcholine (32: 0)+H] was more abundant (P

Embryo production may be enhanced when associated with cows selected on the basis of fertility markers, which should be easy to measure, such as antral follicle count (AFC) and genital tract morphometrics. The objective was to evaluate the effects of AFC class on oocyte 24-h outcome and in vitro embryo production in Bos indicus beef cows. Brahman (n = 151) cows (2-13 years old, 344-803 kg of BW, and 7-9 BCS). Low (LAFC), intermediate (IAFC), and high (HAFC) antral follicle classes were defined as follows: LAFC ≤ 30; IAFC 30-49; and HAFC ≥50 AFC. All follicles ≥3 mm in diameter were aspirated by conventional ovum pick-up technique. Only cumulus–oocyte complexes with at least 2 layers of granulosa cells and homogeneous cytoplasm were used for in vitro culture. They were matured in TCM-199 plus supplements for 24 h at 38.7°C in a 5% CO2 humidified atmosphere. After 24 h of maturation, a subset of oocytes (n = 319) was fixed and analysed under fluorescent microscopy and oocyte outcome was evaluated by classification, as follows: germinal vesicle (GV), germinal vesicle breakdown (GVBD), metaphase I/anaphase I/telophase I (MIAITI), and metaphase II (MII). The second subset of oocytes (n = 797) was fertilized in Ferti-TALP (10-15 oocytes per 60-µL drop) with frozen–thawed semen (18-22 h at 38.7°C in 5% CO2 after Percoll) from a single bull previously tested for good in vitro fertility. Presumptive zygotes were cultivated in CR2 medium for 48 h at 37.8°C in 5% CO2. For the remaining 96 h, embryos were transferred to 10% FCS-supplemented TCM-199 drops until the final evaluation. Data were analysed by the GENMOD, GLM, and CORR procedures of SAS (SAS Institute Inc., Cary, NC, USA). Viable oocytes, total embryos, and embryo production efficiency (viable oocyte/total embryos produced; P