Influence of age and ovarian antral follicle count on the reproductive characteristics of embryo donor mares.

BACKGROUND: Age and antral follicle count (AFC) are related to fertility in cattle, but this information remains limited in mares. METHODS: To verify the influence of age and AFC on the reproductive characteristics of mares, 15 Quarter Horse donors, with 5-15 antral follicles, ranging from three to 17 years old, healthy and in good nutritional status, were divided into groups with low AFC (≤9 follicles) or high AFC (≥10 follicles) and mares considered young (≤9 years) or old (≥10 years). Mares were submitted to ultrasonography to determine the dominant follicle diameter, follicular growth rate, degree of uterine oedema and embryonic recovery for a minimum of three oestrous cycles. RESULTS: AFC was higher (P=0.001) in young mares compared with old mares. An interaction (P=0.001) between AFC and age was observed with regard to follicular growth rate, being that mares with low AFC and are old showed the lowest follicular growth compared with those of low AFC and young, high AFC and old, and high AFC and young. Younger mares and those with high AFC exhibited higher degree of uterine oedema (P<0.05) on the third day of oestrus compared with older mares and with low AFC (3 and 2, respectively). However, in both groups, the degree of oedema reduced by the time of ovulation. CONCLUSION: Follicular growth rate, degree of uterine oedema and the number of antral follicles are higher in young mares with high AFC. However, the rate of embryonic recovery is not influenced by donors' age or AFC.

Growth and Differentiation Factor-9 Supplementation Affects Viability and Morphology of Preantral Follicles in Equine Ovarian Fragments During Short-term in vitro Culture

Abstract This study aimed to evaluate different concentrations of growth and differentiation factor-9 (GDF-9) on the development and maintenance of equine preantral follicle morphology during short-term in vitro culture. Ovaries (n=5) from five mares were collected from a local slaughterhouse and transported to the laboratory, where nine fragments (5x5x1mm) were procured from each ovary. One fragment from each was immediately fixed and submitted for histological analysis (control group; D0). The other eight fragments were cultured in situ for two (D2) or six (D6) days in MEM+ or MEM+ supplemented with GDF-9 at different concentrations (i.e., 50, 100 and 200 ng/mL the GDF-9). After culturing with different concentrations of GDF-9 for 2 or 6 days, the fragments were processed for histological analysis. After two days of cultivation, we observed an increase in the percentage of developing follicles for 0 (MEM+), 50, 100 and 200 ng/mL GDF-9 compared to control (D0; P<0.05). When we evaluated all treatments that preserved follicular integrity, the GDF-9 concentration of 100 ng/mL presented results superior to those of the other cultures (P<0.05). While, at six days of culture, the concentration of 200 ng/mL of GDF-9 appeared to be more efficient in providing development compared to MEM+ (P<0.05). The percentage of morphologically intact follicles in the 6 days culture samples treated with 50 ng/mL of GDF-9 indicated that this concentration was effective in maintaining the integrity of the follicle (P<0.05). We conclude, therefore, that graduated GDF-9 addition to the medium ensure follicular development and is sufficient maintain the architecture.

The correlation between the number of antral follicles and ovarian reserves (preantral follicles) in purebred Bos indicus and Bos taurus cows.

The objectives of this study were to compare populations of preantral follicles between purebred Bos indicus and Bos taurus cows with high or low antral follicle counts (AFC) and to correlate the number of preantral follicles with the population of antral follicles. Nelore (Bos indicus, n=100) and Angus (Bos taurus, n=100) cow ovaries were collected at abattoirs and examined using ultrasonography. Antral follicles ≥3mm were counted, and the cows ovaries were assigned to high (G-High) or low (G-Low) AFC groups based on the mean number (±1 SD) of ovarian antral follicles: Bos indicus with high AFC (≥57 follicles, n=8) or low AFC (≤21 follicles, n=8) and Bos taurus with high (≥45 follicles, n=10) or low AFC (≤13 follicles, n=10). The ovaries were processed, and the number of preantral follicles was estimated. Between-groups comparisons were performed using a Kruskal-Wallis test, and the correlation between preantral and antral follicles was evaluated using a Pearson's correlation test (P≤0.05). A large variation in the number of preantral follicles was observed among the animals. Although there was a correlation between the population of preantral follicles and the number of antral follicles, there was no difference between the mean number of preantral follicles in the Bos indicus G-High (48,349±30,149) and G-Low groups (33,037±31,710) or between the Bos taurus G-High (35,050±36,060) and G-Low groups (30,481±43,360). Therefore, the preantral follicle population did not differ between purebred Bos indicus and Bos taurus cattle with high or low AFC but was correlated with the number of antral follicles. In addition to the large within-groups variation in the number of preantral follicles, some cows with high AFC had lower populations of preantral follicles compared to the low AFC group, and the highest population of preantral follicles was observed in both Bos indicus and Bos taurus with low AFC.

Antral follicle count in cattle: advantages, challenges, and controversy

The antral follicle count (AFC) represents the number of follicles visualized by ultrasonography in the ovaries. Antral follicle count tends to be variable among cows but with high repeatability within the same individual. In the last decade, AFC has been considered a biological marker of fertility with many positive aspects of reproductive efficiency for those animals with high AFC. For instance, a larger number and better-quality embryos, better pregnancy rates, increased concentrations of circulating progesterone, in addition to other characteristics linked to fertility, are all responses observed in individuals with high compared with those with low AFC in Bos taurus cattle. However, the positive association between AFC and fertility did not follow the same pattern for Bosindicus. Recent articles showed no connection between fertility and number of antral follicles or better reproductive efficiency for high AFC group in Zebu cows. Thus, the aim of this review is to discuss the various data concerning AFC between indicus and taurus cattle. Additionally, we consider AFC to be a possible tool to improve cattle performance in reproductive biotechnology.

Antral follicle count in cattle: advantages, challenges, and controversy

The antral follicle count (AFC) represents the number of follicles visualized by ultrasonography in the ovaries. Antral follicle count tends to be variable among cows but with high repeatability within the same individual. In the last decade, AFC has been considered a biological marker of fertility with many positive aspects of reproductive efficiency for those animals with high AFC. For instance, a larger number and better-quality embryos, better pregnancy rates, increased concentrations of circulating progesterone, in addition to other characteristics linked to fertility, are all responses observed in individuals with high compared with those with low AFC in Bos taurus cattle. However, the positive association between AFC and fertility did not follow the same pattern for Bosindicus. Recent articles showed no connection between fertility and number of antral follicles or better reproductive efficiency for high AFC group in Zebu cows. Thus, the aim of this review is to discuss the various data concerning AFC between indicus and taurus cattle. Additionally, we consider AFC to be a possible tool to improve cattle performance in reproductive biotechnology.(AU)

Effect of follicular diameter, time of first cleavage and H3K4 methylation on embryo production rates of Bos indicus cattle / Efeito do diâmetro folicular, momento da primeira clivagem e metilação da H3K4 na produção embrionária de vacas Bos indicus

This study aimed investigate the relationship between epigenetics, follicular diameter and cleavage speed, by evaluating the developmental potential and occurence of H3K4 monomethylation of early-, intermediate- and late-cleaving Bos indicus embryos from in vitro fertilized oocytes originating from follicles up to 2 mm in diameter or between 4 and 8 mm in diameter. Oocytes (n = 699) from small follicles (? 2 mm) and 639 oocytes from large follicles (4-8 mm) were punched from 1,982 Bos indicus slaughterhouse ovaries. After maturation and in vitro fertilization (IVF), the cultured embryos were separated into early (? 28 h post-IVF), intermediate (> 28 h and ? 34 h post-IVF) and late (> 34 h and ? 54 h post-IVF) cleavage groups. Blastocysts were subjected to an immunofluorescence assessment for H3K4me investigation. The blastocyst rate for large follicles (36.3%) was higher than that for small follicles (22.9%, P 0.05). In addition, blastocyst rates for early and intermediate cleavage groups (45.3% and 33.8%, respectively) were higher than that for late cleavage group (13.5%, P 0.05). The blastocysts from all groups displayed H3K4me staining by immunofluorescence, particularly intense in what seemed to be trophectoderm cells and weak or absent in cells seemingly from the inner cell mass. For the first time for indicus embryos, data from this study demonstrate that higher blastocyst embryorates are obtained from embryos that cleave within 34 h after fertilization and from those produced fromfollicles of 4-8 mm in diameter, indicating a greater ability of these embryos to develop to the stage ofembryonic preimplantation. This is the first article demonstrating the occurrence of H3K4me in cattleembryos; its presence in all the evaluated blastocysts suggests that this histone modification plays a keyrole in maintaining embryo viability at preimplantation stage.

Este estudo teve como objetivo investigar a relação entre a epigenética, o diâmetro folicular e a velocidade de clivagem, avaliando o potencial de desenvolvimento e a ocorrência de monometilação da H3K4 em embriões Bos indicus de clivagem precoce, intermediária e tardia produzidos a partir de oócitos fertilizados in vitro oriundos de folículos de até 2 mm de diâmetro ou entre 4 e 8 mm de diâmetro. Oócitos (n = 699) de folículos pequenos (? 2 mm) e 639 oócitos de folículos grandes (4-8 mm) foram puncionados de 1982 ovários de vacas Bos indicus de abatedouro. Após a maturação e fertilização in vitro (FIV), os embriões cultivados foram separados nos grupos de clivagem precoce (? 28 h pós-FIV), intermediária (> 28 h e ? 34 h pós-FIV) e tardia (> 34 h e ? 54 h pós-FIV). Os blastocistos foram submetidos à imunofluorescência para investigação de H3K4me. A taxa de blastocisto para embriões provenientes de folículos grandes (36,3%) foi maior que de folículos pequenos (22,9%; p 0,05). Ainda, as taxas de blastocisto para os grupos de clivagem precoce e intermediária (45,3% e 33,8%, respectivamente) foram maiores que para o grupo de clivagem tardia (13,5%; p 0,05). Blastocistos de todos os grupos mostraram marcação para H3K4me à imunofluorescência, particularmente intensa no que pareciam ser células do trofectoderma e fraca ou ausente em células semelhantes às da massa massa celular interna. Pela primeira vez em embriões indicus, os dados deste estudo demonstram que maiores taxas deblastocisto são obtidas de embriões que clivam em até 34 h pós-fertilização e dos oriundos de folículos de 4 a 8 mm de diâmetro, indicando uma maior habilidade desses embriões de se desenvolverem até o estágio de pré-implantação embrionária.

Effect of follicular diameter, time of first cleavage and H3K4 methylation on embryo production rates of Bos indicus cattle / Efeito do diâmetro folicular, momento da primeira clivagem e metilação da H3K4 na produção embrionária de vacas Bos indicus

This study aimed investigate the relationship between epigenetics, follicular diameter and cleavage speed, by evaluating the developmental potential and occurence of H3K4 monomethylation of early-, intermediate- and late-cleaving Bos indicus embryos from in vitro fertilized oocytes originating from follicles up to 2 mm in diameter or between 4 and 8 mm in diameter. Oocytes (n = 699) from small follicles (? 2 mm) and 639 oocytes from large follicles (4-8 mm) were punched from 1,982 Bos indicus slaughterhouse ovaries. After maturation and in vitro fertilization (IVF), the cultured embryos were separated into early (? 28 h post-IVF), intermediate (> 28 h and ? 34 h post-IVF) and late (> 34 h and ? 54 h post-IVF) cleavage groups. Blastocysts were subjected to an immunofluorescence assessment for H3K4me investigation. The blastocyst rate for large follicles (36.3%) was higher than that for small follicles (22.9%, P 0.05). In addition, blastocyst rates for early and intermediate cleavage groups (45.3% and 33.8%, respectively) were higher than that for late cleavage group (13.5%, P 0.05). The blastocysts from all groups displayed H3K4me staining by immunofluorescence, particularly intense in what seemed to be trophectoderm cells and weak or absent in cells seemingly from the inner cell mass. For the first time for indicus embryos, data from this study demonstrate that higher blastocyst embryorates are obtained from embryos that cleave within 34 h after fertilization and from those produced fromfollicles of 4-8 mm in diameter, indicating a greater ability of these embryos to develop to the stage ofembryonic preimplantation. This is the first article demonstrating the occurrence of H3K4me in cattleembryos; its presence in all the evaluated blastocysts suggests that this histone modification plays a keyrole in maintaining embryo viability at preimplantation stage.(AU)

Este estudo teve como objetivo investigar a relação entre a epigenética, o diâmetro folicular e a velocidade de clivagem, avaliando o potencial de desenvolvimento e a ocorrência de monometilação da H3K4 em embriões Bos indicus de clivagem precoce, intermediária e tardia produzidos a partir de oócitos fertilizados in vitro oriundos de folículos de até 2 mm de diâmetro ou entre 4 e 8 mm de diâmetro. Oócitos (n = 699) de folículos pequenos (? 2 mm) e 639 oócitos de folículos grandes (4-8 mm) foram puncionados de 1982 ovários de vacas Bos indicus de abatedouro. Após a maturação e fertilização in vitro (FIV), os embriões cultivados foram separados nos grupos de clivagem precoce (? 28 h pós-FIV), intermediária (> 28 h e ? 34 h pós-FIV) e tardia (> 34 h e ? 54 h pós-FIV). Os blastocistos foram submetidos à imunofluorescência para investigação de H3K4me. A taxa de blastocisto para embriões provenientes de folículos grandes (36,3%) foi maior que de folículos pequenos (22,9%; p 0,05). Ainda, as taxas de blastocisto para os grupos de clivagem precoce e intermediária (45,3% e 33,8%, respectivamente) foram maiores que para o grupo de clivagem tardia (13,5%; p 0,05). Blastocistos de todos os grupos mostraram marcação para H3K4me à imunofluorescência, particularmente intensa no que pareciam ser células do trofectoderma e fraca ou ausente em células semelhantes às da massa massa celular interna. Pela primeira vez em embriões indicus, os dados deste estudo demonstram que maiores taxas deblastocisto são obtidas de embriões que clivam em até 34 h pós-fertilização e dos oriundos de folículos de 4 a 8 mm de diâmetro, indicando uma maior habilidade desses embriões de se desenvolverem até o estágio de pré-implantação embrionária. (AU)

Considerações sobre o uso da crioterapia como método preventivo da laminite aguda em equinos / Considerations on the use of cryotherapy as a preventive method of acute laminitis in horse / Consideraciones para el uso de crioterapia cómo método de prevención de la laminitis aguda en los caballos

A laminite é uma doença inflamatória das lâminas internas do casco, a qual acarreta na degeneração de estruturas de sustentação da terceira falange. Esta possui três fases, de acordo com a sua evolução, a saber: a fase de desenvolvimento, a fase aguda, e por fim, a fase crônica, A crioterapia foi instituída com o intuito de prevenir ou minimizar o processo inflamatório presente na primeira fase da doença. Este método terapêutico tem como finalidade proporcionar certo grau de analgesia, diminuir o metabolismo local, levando a uma resposta vascular, de modo a diminuir a chegada dos mediadores do processo inflamatório aos tecidos. Para que o método apresente resultado satisfatório, é necessário o conhecimento sobre a técnica e o uso de equipamentos adequados, a fim de que haja uma resposta eficiente.(AU)

Laminitis is an inflammatory disease of the internal laminae of the hoof, which leads to degeneration of support structures of the third phalanx. According to its evolution, this disease has three phases: the first corresponds to the development phase, the second one, is the acute phase, and finally, the chronic phase of the disease. The cryotherapy was instituted with the purpose of preventing or minimizing the inflammatory process present in the first phase of the disease. This therapeutic method aims to provide a certain degree of analgesia, decrease local metabolism, leading to a vascular response, so as to decrease the influx of inflammatory mediators to the tissues. In order to present a satisfactory result from this the method, it is necessary knowledge about the technique and the use of appropriate equipment, so than, an efficient response.(AU)

La cojera es una enfermedad inflamatoria de las palas interiores del casco, que causa la degeneración de las estructuras de soporte de la terceros falange. Esto tiene tres fases, de acuerdo con su desarrollo, a saber: etapa de desarrollo, la fase aguda, y finalmente la fase crónica. La crioterapia era instituida el fin de evitar o reducir al mínimo el proceso inflamatorio en la primera etapa de la enfermedad. Este método terapéutico está destinado a proporcionar un cierto grado de analgesia, y la reducción del metabolismo local, lo que lleva a una respuesta vascular, a fin de reducir la llegada de mediadores de la inflamación de los tejidos. Para el resultado satisfactorio del método, el conocimiento de la técnica y el uso de equipo adecuado son necesarios, de modo que hay una respuesta eficiente.(AU)

Considerações sobre o uso da crioterapia como método preventivo da laminite aguda em equinos / Considerations on the use of cryotherapy as a preventive method of acute laminitis in horse / Consideraciones para el uso de crioterapia cómo método de prevención de la laminitis aguda en los caballos

A laminite é uma doença inflamatória das lâminas internas do casco, a qual acarreta na degeneração de estruturas de sustentação da terceira falange. Esta possui três fases, de acordo com a sua evolução, a saber: a fase de desenvolvimento, a fase aguda, e por fim, a fase crônica, A crioterapia foi instituída com o intuito de prevenir ou minimizar o processo inflamatório presente na primeira fase da doença. Este método terapêutico tem como finalidade proporcionar certo grau de analgesia, diminuir o metabolismo local, levando a uma resposta vascular, de modo a diminuir a chegada dos mediadores do processo inflamatório aos tecidos. Para que o método apresente resultado satisfatório, é necessário o conhecimento sobre a técnica e o uso de equipamentos adequados, a fim de que haja uma resposta eficiente.

Laminitis is an inflammatory disease of the internal laminae of the hoof, which leads to degeneration of support structures of the third phalanx. According to its evolution, this disease has three phases: the first corresponds to the development phase, the second one, is the acute phase, and finally, the chronic phase of the disease. The cryotherapy was instituted with the purpose of preventing or minimizing the inflammatory process present in the first phase of the disease. This therapeutic method aims to provide a certain degree of analgesia, decrease local metabolism, leading to a vascular response, so as to decrease the influx of inflammatory mediators to the tissues. In order to present a satisfactory result from this the method, it is necessary knowledge about the technique and the use of appropriate equipment, so than, an efficient response.

La cojera es una enfermedad inflamatoria de las palas interiores del casco, que causa la degeneración de las estructuras de soporte de la terceros falange. Esto tiene tres fases, de acuerdo con su desarrollo, a saber: etapa de desarrollo, la fase aguda, y finalmente la fase crónica. La crioterapia era instituida el fin de evitar o reducir al mínimo el proceso inflamatorio en la primera etapa de la enfermedad. Este método terapéutico está destinado a proporcionar un cierto grado de analgesia, y la reducción del metabolismo local, lo que lleva a una respuesta vascular, a fin de reducir la llegada de mediadores de la inflamación de los tejidos. Para el resultado satisfactorio del método, el conocimiento de la técnica y el uso de equipo adecuado son necesarios, de modo que hay una respuesta eficiente.