Peroxisome proliferator-activated receptor delta-PPARδ agonist (L-165041) enhances bovine embryo survival and post vitrification viability.

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.

Antral follicular count and its relationship with ovarian volume, preantral follicle population and survival, oocyte meiotic progression and ultrastructure of in vitro matured bovine cumulus-oocyte complexes.

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 (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.

In vitro performance of Zebu (Bos indicus) and Taurus (Bos taurus) donor cow embryos / Desempenho de vacas doadoras zebuínas (Bos indicus) e taurinas (Bos taurus) na produção de embrião in vitro

In this study, the in vitro production of bovine embryos from zebu and taurine donors was compared. Cumulus-oocyte complexes (COCs) were obtained from 167 Bos taurus and 161 Bos indicus donors by ovum pick-up. COCs were classified based on their morphological quality, matured in incubators for 22 to 24 h in maturation medium, and then fertilized for 18 to 22 h. The zygotes were transferred to the culture medium for seven days. The embryos were classified as morula (OM), initial blastocyst (BI), blastocyst (BL), and expanded blastocyst (BX), before being transferred to synchronized recipient cows. Pregnancy was diagnosed 30-45 days post-transfer. The Bos indicus donors had a higher oocyte yield (n = 2556) than Bos taurus donors (n = 1903) (P = 0.008). The COCs from zebu donors had a better morphological quality than those from taurine donors (n = 689 vs. 444 for grade 1 COC, P < 0.0001; n = 681 vs. 509 for grade 2 COC, P = 0.010, for zebu and taurine donors, respectively). There were differences in embryo production percentages obtained from OM (0.44% from zebu and 6.42% from taurine, P = 0.017), BL (14.18% from zebu and 3.74% from taurine, P < 0.0001), and BX (81.43% from zebu and 75.13% from taurine, P < 0.0001). No significant difference was observed for embryo production from BI and pregnancy rate (P > 0.05). The Bos indicus cows showed greater oocyte recovery, number of viable oocytes, and production of viable embryos than the Bos taurus cows.(AU)

O objetivo foi avaliar a produção in vitro de embriões bovinos a partir de doadoras zebuína e taurina. Os complexos cumulus oócitos (COCs) foram obtidos de 167 doadoras Bos taurus e 161 Bos indicus, por meio de Ovum pick-up. Os COCs foram classificados quanto à qualidade morfológica, maturados em incubadora por 22 a 24h em meio de maturação, e encaminhados à fertilização entre 18 e 22h. Os zigotos foram transferidos para meio de cultivo, onde permaneceram durante sete dias. Os embriões foram classificados em mórula (MO), blastocisto inicial (BI), blastocisto (BL) e blastocisto expandido (BX), envasados em palhetas contendo meio de cultivo, e inovulados em receptoras sincronizadas. Após 30 e 45 dias da inovulação, realizou-se o diagnóstico de gestação. Os animais Bos indicus apresentaram maior produção oocitária (n = 2556) em comparação aos Bos taurus (n = 1903) (P = 0,008). Os COCs das doadoras zebu apresentaram melhor qualidade morfológica do que das doadoras taurina (n = 689 vs. 444 para COCs grau I, P < 0,0001; n = 681 vs. 509 para COCs grau II, P = 0,010, para doadoras zebu e taurina, respectivamente). Quanto à produção de embriões, houve diferença nas porcentagens obtidas de MO (0,44% zebuínas e 6,42% taurinas, P = 0,017), BL (14,18% zebuínas e 3,74% taurinas, P < 0,0001) e BX (81,43% zebuínas e 75,13% taurinas, P < 0,0001). Não foi observado diferença para BI e taxa de prenhez (P > 0,05). Os animais Bos indicus apresentaram maior recuperação oocitária e maior número de oócitos viáveis em comparação às vacas Bos taurus, como também, maior produção de embriões viáveis.(AU)

Effects of vulvar width and antral follicle count on oocyte quality, in vitro embryo production and pregnancy rate in Bos taurus taurus and Bos taurus indicus cows.

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 and <24) 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 (P < 0.05) 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.

In vitro performance of Zebu (Bos indicus) and Taurus (Bos taurus) donor cow embryos / Desempenho de vacas doadoras zebuínas (Bos indicus) e taurinas (Bos taurus) na produção de embrião in vitro

In this study, the in vitro production of bovine embryos from zebu and taurine donors was compared. Cumulus-oocyte complexes (COCs) were obtained from 167 Bos taurus and 161 Bos indicus donors by ovum pick-up. COCs were classified based on their morphological quality, matured in incubators for 22 to 24 h in maturation medium, and then fertilized for 18 to 22 h. The zygotes were transferred to the culture medium for seven days. The embryos were classified as morula (OM), initial blastocyst (BI), blastocyst (BL), and expanded blastocyst (BX), before being transferred to synchronized recipient cows. Pregnancy was diagnosed 30-45 days post-transfer. The Bos indicus donors had a higher oocyte yield (n = 2556) than Bos taurus donors (n = 1903) (P = 0.008). The COCs from zebu donors had a better morphological quality than those from taurine donors (n = 689 vs. 444 for grade 1 COC, P 0.05). The Bos indicus cows showed greater oocyte recovery, number of viable oocytes, and production of viable embryos than the Bos taurus cows.

O objetivo foi avaliar a produção in vitro de embriões bovinos a partir de doadoras zebuína e taurina. Os complexos cumulus oócitos (COC’s) foram obtidos de 167 doadoras Bos taurus e 161 Bos indicus, por meio de Ovum pick-up. Os COC’s foram classificados quanto à qualidade morfológica, maturados em incubadora por 22 a 24h em meio de maturação, e encaminhados à fertilização entre 18 e 22h. Os zigotos foram transferidos para meio de cultivo, onde permaneceram durante sete dias. Os embriões foram classificados em mórula (MO), blastocisto inicial (BI), blastocisto (BL) e blastocisto expandido (BX), envasados em palhetas contendo meio de cultivo, e inovulados em receptoras sincronizadas. Após 30 e 45 dias da inovulação, realizou-se o diagnóstico de gestação. Os animais Bos indicus apresentaram maior produção oocitária (n = 2556) em comparação aos Bos taurus (n = 1903) (P = 0,008). Os COCs das doadoras zebu apresentaram melhor qualidade morfológica do que das doadoras taurina (n = 689 vs. 444 para COCs grau I, P 0,05). Os animais Bos indicus apresentaram maior recuperação oocitária e maior número de oócitos viáveis em comparação às vacas Bos taurus, como também, maior produção de embriões viáveis.

Effects of melatonin on morphology and development of primordial follicles during in vitro culture of bovine ovarian tissue.

This study aims to investigate the effect of melatonin on activation, growth and morphology of bovine primordial follicles, as well as on stromal cells density in ovarian tissues after in vitro culture. Ovarian fragments were cultured in α-MEM+ alone or supplemented with melatonin (250, 500, 1,000 or 2,000 pM) for a period of six days. Non-cultured and cultured tissues were processed for histological analysis; according to developmental stages, follicles were classified as primordial or growing follicles. These follicles were further classified as morphologically normal or degenerated. Ovarian stromal cell density was also evaluated. The percentages of primordial and developing follicles, as well as those classified of normal follicles, were compared by Fisher's exact test, and the differences were considered significant when p < .05. The results showed that the presence of 1,000 and 2,000 pM melatonin in culture medium promoted a reduction in the percentage of primordial follicles and an increase in the percentage of development follicles, when compared to follicles cultured in control medium. On the other hand, the presence of 250 or 500 pM melatonin did not show a significant effect on the percentage of primordial and developing follicles. Besides that, the presence of 500, 1,000 and 2,000 pM melatonin maintained the percentage of normal follicles similar to those seen uncultured control. Moreover, tissues cultured in presence of 1,000 pM melatonin showed a higher percentage of normal follicles when compared to follicles cultured in the presence of 250 pM melatonin. It was observed a similar profile of stromal density in both uncultured tissues and those cultured in vitro in the presence of melatonin. In conclusion, melatonin (1,000 and 2,000 pM) promotes bovine primordial follicles activation and maintains the stromal cell density during in vitro culture of ovarian cortical tissue.

Anti-Müllerian Hormone (AMH), antral follicle count (AFC), external morphometrics and fertility in Tabapuã cows.

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 Tabapuã (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 (<28), intermediate (28-50) and greater (>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 ±â€¯23.96) primiparous (47.54 ±â€¯26.16) and multiparous (41.08 ±â€¯25) cows and was negatively correlated with pregnancy interval (PI), such that, as PI decreased (r = -0.28; P < 0.005), AFC increased. Vulva width was not affected (P = 0.08) by parity and was larger (P < 0.05) for females in the greater AFC class (8.81 ±â€¯0.12 cm) compared with the intermediate (8.42 ±â€¯0.11 cm) and lesser (8.38 ±â€¯0.13 cm) classes. As vulva width increased, parturition interval decreased (r = -0.15; P < 0.005) and overall reproductive efficiency increased (r = 0.17, p < 0.005). Thoracic depth was associated with greater AFCs (r = 0.10; P < 0.001). Anti-Müllerian hormone concentrations were greater (P < 0.05) for animals in the greater AFC class (1.15 ±â€¯0.09 ng/mL) compared with the lesser (0.44 ±â€¯0.02 ng/mL) and intermediate (0.73 ±â€¯0.05 ng/mL) classes. Vulva width, AFC, external morphometrics and AMH concentrations were moderately associated, considering the low heritability of fertility traits, and should be studied further to be considered in the selection for fertility in Bos indicus cattle.

Detecção eletrônica do estro em vacas leiteiras: uma revisão / Detection electronic of estrus in dairy cows: a review

Estrus detection efficiency is fundamental to provide viability in animal production, with effect on the calving interval (IEP), fertility and conception rate in herds submitted to artificial insemination (AI). This condition must be sought, because the activities related to reproduction are much more intense and failures would lead to serious problems in management, with major economic repercussions. The objective of this review of the literature about the electronic detection of estrus in dairy cows, addressing physical activity gauges, like the pedometer and collar; mounts detectors with pressure sensitivity, as the radio telemetry; electrical impedance measurement of vaginal mucus; video cameras; and method using biosensors to quantify the progesterone in milk. It can therefore be concluded that these electronic methods of detection of estrus have great effectiveness, especially when they are associated with the visual observation method of estrus detection. Among all methods, it was noted, in this study, that the pedometer is the most widely used and most effective method. The radiotelemetry, stands out for monitoring the activity of assemble, registering the time and its duration, is very efficient and can be used to indicate the beginning and end of estrus, showing the best time to perform the artificial insemination.(AU)

A eficiência na detecção do estro é fundamental para proporcionar a viabilidade na produção animal, com efeito sobre o intervalo de partos (IEP), taxa de concepção e de fertilidade em rebanhos submetidos à inseminação artificial (IA). Essa condição deve ser buscada, pois as atividades relacionadas à reprodução são muito mais intensas e falhas levariam a graves problemas no manejo, com grandes repercussões econômicas. Objetivou-se apresentar uma revisão da literatura sobre a detecção eletrônica de estro em vacas leiteiras, abordando os medidores de atividade física, como o pedômetro e o colar; detectores de monta com sensibilidade à pressão, como a radiotelemetria; medição da impedância elétrica do muco vaginal; câmeras de vídeo; e método utilizando biossensores para quantificar a progesterona no leite. Pode-se concluir que esses métodos eletrônicos de detecção de estro possuem grande eficácia, principalmente quando estão associados ao método de observação visual de detecção de estro. Dentre todos os métodos, constatou-se, neste estudo, que o pedômetro é o mais utilizado e mais eficaz. A radiotelemetria destaca-se por monitorar a atividade da monta, registrando o horário e a sua duração; é bastante eficiente e pode ser usada para indicar o início e final do estro, mostrando o melhor momento para realizar a inseminação artificial.(AU)

Detecção eletrônica do estro em vacas leiteiras: uma revisão / Detection electronic of estrus in dairy cows: a review

Estrus detection efficiency is fundamental to provide viability in animal production, with effect on the calving interval (IEP), fertility and conception rate in herds submitted to artificial insemination (AI). This condition must be sought, because the activities related to reproduction are much more intense and failures would lead to serious problems in management, with major economic repercussions. The objective of this review of the literature about the electronic detection of estrus in dairy cows, addressing physical activity gauges, like the pedometer and collar; mounts detectors with pressure sensitivity, as the radio telemetry; electrical impedance measurement of vaginal mucus; video cameras; and method using biosensors to quantify the progesterone in milk. It can therefore be concluded that these electronic methods of detection of estrus have great effectiveness, especially when they are associated with the visual observation method of estrus detection. Among all methods, it was noted, in this study, that the pedometer is the most widely used and most effective method. The radiotelemetry, stands out for monitoring the activity of assemble, registering the time and its duration, is very efficient and can be used to indicate the beginning and end of estrus, showing the best time to perform the artificial insemination.

A eficiência na detecção do estro é fundamental para proporcionar a viabilidade na produção animal, com efeito sobre o intervalo de partos (IEP), taxa de concepção e de fertilidade em rebanhos submetidos à inseminação artificial (IA). Essa condição deve ser buscada, pois as atividades relacionadas à reprodução são muito mais intensas e falhas levariam a graves problemas no manejo, com grandes repercussões econômicas. Objetivou-se apresentar uma revisão da literatura sobre a detecção eletrônica de estro em vacas leiteiras, abordando os medidores de atividade física, como o pedômetro e o colar; detectores de monta com sensibilidade à pressão, como a radiotelemetria; medição da impedância elétrica do muco vaginal; câmeras de vídeo; e método utilizando biossensores para quantificar a progesterona no leite. Pode-se concluir que esses métodos eletrônicos de detecção de estro possuem grande eficácia, principalmente quando estão associados ao método de observação visual de detecção de estro. Dentre todos os métodos, constatou-se, neste estudo, que o pedômetro é o mais utilizado e mais eficaz. A radiotelemetria destaca-se por monitorar a atividade da monta, registrando o horário e a sua duração; é bastante eficiente e pode ser usada para indicar o início e final do estro, mostrando o melhor momento para realizar a inseminação artificial.