Influência da estação do ano nas características do sêmen e na concentração de hormônios em touros Nelore e Simental

Estudou-se a influência das quatro estações do ano nas características do sêmen e nas concentrações de testosterona e cortisol em touros. Cinco touros Nelore e cinco Simental entre 48 e 72 meses de idade, criados extensivamente, foram avaliados andrologicamente por meio de exames físicos das características morfológicas do sêmen e das concentrações séricas de testosterona e cortisol. Houve redução na motilidade e no vigor do sêmen no inverno (P<0,05) na raça Simental. Observou-se correlação (P<0,01) entre testosterona x motilidade (0,69) e testosterona x vigor (0,57) na raça Simental, e cortisol x motilidade (0,68) e cortisol x vigor (0,65) na raça Nelore. O efeito das estações do ano modificou a qualidade do sêmen com aumento da motilidade e vigor espermáticos na primavera e verão nos touros Simental. A concentração de cortisol diminuiu no outono nos touros Nelore.

Abordagem multivariada envolvendo características físicas e morfológicas do sêmen bovino, idade dos touros e época de colheita de sêmen

Avaliaram-se as relações entre características físicas e morfológicas do sêmen de bovinos das subespécies Bos taurus taurus e Bos taurus indicus com a idade dos touros e a época de colheita do sêmen. Utilizaram-se observações feitas durante o período de 1993 a 1999, em 42 touros com 12 a 174 meses de idade, divididos em cinco classes: Bos taurus taurus - 12 a 36 meses; 37 a 60 meses; 61 a 84 meses; 85 a 108 meses; e 109 a 138 meses; e Bos taurus indicus - 12 a 42 meses; 43 a 72 meses; 73 a 102 meses; 103 a 132 meses; e 133 a 174 meses. As características analisadas foram: volume, turbilhonamento espermático, concentração espermática, motilidade espermática, vigor espermático, anormalidades espermáticas e integridade do acrossoma. No processamento das análises, utilizaram-se as médias das características em cada classe de idade e o mês de colheita de sêmen durante o período de 1993 a 1999. As técnicas estatísticas multivariadas de componentes principais e de agrupamento hierárquico mostraram resultados que podem contribuir na escolha de sêmen de melhor qualidade. As classes 103 a 132 meses e 133 a 174 meses para a subespécie Bos taurus indicus e a classe de idade 109 a 138 meses para a subespécie Bos taurus taurus foram as mais contrastantes. Para ambas as subespécies, nos meses mais úmidos, o sêmen apresentou menor qualidade, principalmente na subespécie Bos taurus taurus. As características do sêmen mais contrastantes no estudo por classes e no período de coleta de sêmen foram: vigor espermático, motilidade espermática, concentração espermática, integridade do acrossoma e anormalidades terciárias no sêmen.

Fecundação in vitro de ovócitos bovinos com sêmen submetido a diferentes diluidores

O objetivo deste trabalho foi avaliar a influência do diluidor do sêmen no desenvolvimento in vitro de ovócitos bovinos após a maturação e fecundação in vitro. Ejaculado de um reprodutor foi fracionado e submetido a três diluidores: Lactose/gema de ovo (LG), Citrato/gema de ovo (CG) e Tris/gema de ovo (TG). Amostras deste material foram envasadas, congeladas e estocadas em N² e, posteriormente, descongeladas; a fração móvel foi separada por gradiente descontínuo de Percoll. A concentração espermática foi ajustada para 10 x 10(6)/mL e a capacitação espermática, induzida com 10 µg/mL de heparina. Após 24 horas de cultura para maturação in vitro, os ovócitos, aspirados de folículos ovarianos, foram inseminados com sêmen diluído em meio TALP e, após 48 horas de cultura, os zigotos foram transferidos para gotas de meio TCM 199, com 5% de soro fetal bovino, 5% de soro de vaca em estro e suspensão de células epiteliais do oviduto bovino, cobertas com óleo de silicone, e mantidos em cultura por nove dias. Todas as culturas foram realizadas a 38,5ºC em atmosfera com 5% de CO2. Os dados foram analisados pelo teste do qui-quadrado e houve diferença com relação à taxa de clivagem (TC), sendo as médias de 66,0; 69,3; e 54,4% para LG, CG e TG, respectivamente. Não houve diferença entre tratamentos com relação às taxas de mórulas/blastocistos ou de eclosão. O diluidor do sêmen não teve efeito sobre o desenvolvimento in vitro de embriões bovinos, embora a TC tenha sido afetada.

Effect of sequence of insemination after simultaneous thawing of multiple semen straws on conception rate to timed AI in suckled multiparous Nelore cows

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Chromatin and morphometry alterations of bovine sperm head after Percoll (TM) density gradient centrifugation

The aim of this work was to assess the morphometry and chromatin integrity of bovine sperm head after a three layers discontinuous Percoll (TM) density gradient centrifugation. Frozen semen doses were obtained from six bulls of different breeds, including three taurine and three Zebu animals. Three ejaculates per bull were evaluated. The semen doses were thawed and two smears were made from each sample before (control) and after the Percoll (TM) centrifugation (Percoll (TM) group). The smears were stained with toluidine blue and grayscale digital images were captured and processed in Scilab environment software. It was observed that chromatin heterogeneity was reduced (P<0.05) and chromatin decondensation was increased (P<0.05) after the Percollm treatment utilized. In addition, it was observed that sperm head length was higher (P<0.05) and the side symmetry was lower (P<0.05) in centrifuged sperm cells. When analyzed separately by subspecies, it was observed that the decrease (P<0.05) in chromatin heterogeneity and the increase (P<0.05) in chromatin decondensation occurred in Zebu sperm heads. In addition, the length and the width:length ratio of sperm heads was affected by Percoll (TM) centrifugation in Zebu semen. In conclusion, the three layers discontinuous Percoll (TM) centrifugation increased the chromatin decondensation and the morphometric alterations of frozen-thawed bovine semen. However, the real implication of these findings in fertility rates of centrifuged sperm must be investigated. (C) 2011 Elsevier B.V. All rights reserved.

Effects of discontinuous Percoll gradient centrifugation on the quality of bovine spermatozoa evaluated with computer-assisted semen analysis and fluorescent probes association

The objective of this study was to evaluate the quality of bovine frozen-thawed sperm cells after Percoll gradient centrifugation. Frozen semen doses were obtained from six bulls of different breeds, including three taurine and three Zebu animals. Four ejaculates per bull were evaluated before and after discontinuous Percoll gradient centrifugation. Sperm motility was assessed by computer-assisted semen analysis and the integrity of the plasma and acrosomal membranes, as well as mitochondrial function, were evaluated using a combination of fluorescent probes propidium iodide, fluorescein isothiocyanate-conjugated Pisum sativum agglutinin and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide. The procedure of Percoll gradient centrifugation increased the percentage of total and progressive sperm motility, beat frequency, rectilinear motility, linearity and rapidly moving cells. In addition, the percentage of cells with intact plasma membrane and mitochondrial membrane potential was increased in post-centrifugation samples. However, the percentage of sperm cells with intact acrosomal membrane was markedly reduced. The method used selected the motile cells with intact plasma membrane and higher mitochondrial functionality in frozen-thawed bull semen, but processing, centrifugation and/or the Percoll medium caused damage to the acrosomal membrane.

Detection of Leptospira spp. in the semen and urine of bulls serologically reactive to Leptospira interrogans serovar hardjo

O presente trabalho avaliou a PCR na detecção de leptospiras em sêmen e urina de dez touros sorologicamente reagentes, comparando seus resultados com aqueles obtidos por outras técnicas de diagnóstico. Foram realizadas duas colheitas de materiais em dias alternados. As amostras de sêmen e de urina foram separadas em alíquotas para visualização direta em microscopia de campo escuro, inoculação em hamsters (apenas para o sêmen), isolamento em meio de cultura e PCR. Nenhum hamster apresentou positividade na prova de soroaglutinação microscópica (SAM); fragmentos de rins e fígado desses animais foram utilizados para a tentativa de isolamento em meio de cultura, sendo positivo o cultivo a partir do rim de hamster inoculado com semen de um touro, e do fígado de hamsters inoculados com o semen de três touros. O isolamento em meio de cultura foi negativo para todas as amostras de sêmen, mas foi positivo para cinco amostras de urina. Na PCR não houve resultado positivo para as amostras de sêmen, e apenas uma amostra de urina apresentou resultado positivo, sendo coincidente com uma das culturas positivas. Não foi possível visualizar leptospiras em nenhuma das amostras por exame direto em microscopia de campo escuro.

Efeito da idade do touro e do período de colheita de sêmen sobre as características físicas e morfológicas do sêmen de bovinos de raças européias e zebuínas

Este trabalho foi realizado com o objetivo de avaliar o efeito da idade de touros europeus e zebuínos e do período de colheita do sêmen sobre as características físicas e morfológicas do sêmen desses animais produzido em uma central de inseminação no período de 1993 a 1999. Os dados de produção de sêmen dos touros foram agrupados em cinco classes de idade (12 a 36 meses, 37 a 60 meses, 61 a 84 meses, 85 a 108 meses e 109 a 142 meses) e quatro períodos de colheita (período 1: dezembro a fevereiro; período 2: março a maio; período 3: junho a agosto e período 4: setembro a novembro). As classes de idade determinaram diferenças significativas no volume, turbilhonamento espermático, nas anormalidades primárias, secundárias, terciárias e totais, na integridade de acrossoma e na quantidade média de doses por ejaculado, cujos valores foram maiores nos zebuínos no período 4 (setembro a outubro). As maiores porcentagens de anormalidades totais, nas duas subespécies, foram observadas nos animais mais jovens (12 a 36 meses) e nos mais velhos (109 e 142 meses). Os zebuínos mais velhos produziram sêmen mais concentrado e maiores quantidades médias de doses por ejaculado. em touros europeus, o sêmen menos concentrado e as maiores porcentagens de anormalidades espermáticas foram observadas no período 1 (dezembro a fevereiro), consequentemente, menores quantidades de doses de sêmen por ejaculado foram produzidas por essa subespécie, o que pode ter sido um efeito do estresse calórico sofrido por estes animais antes da colheita de sêmen. A idade e o período de colheita influenciam na qualidade do sêmen de touros doadores mantidos em regime de colheita para comercialização.

Efeito da idade sobre as principais características andrológicas de touros Brangus- Efeito da idade sobre as principais características andrológicas de touros BrangusIbagé criados extensivamente no estado do Mato Grosso do Sul - Brasil Ibagé criados extensivamente no estado do Mato Grosso do Sul - Brasil

Background: Bull fertility is extremely important for beef cattle production systems and has a multiplier impact on the economical and zootechnical indexes of the herd. Bulls raised in tropical conditions may present varied seminal characteristics due to, among other causes, different management practices and quality of pastures. Another factor that influences the semen characteristics is the age of the sire. The reproductive potential of bull evaluated through the andrological exam aims to ensure the semen quality and to improve the herd reproductive efficiency. The aim of this study was to evaluate the main semen parameters of Brangus-Ibage bulls extensively reared in eastern Mato Grosso do Sul state and to verify the effect of age on the andrological characteristics analyzed. It was also evaluated the correlation between age, scrotal circumference, and physical and morphological sperm characteristics produced by the Brangus bulls.Materials, Methods & Results: The study took place in the month of July 2010, during the routine andrological examination of 168 synthetic Brangus-Ibage bulls (5/8 Angus x 3/8 Nelore), belonging to the same property. For data analysis the animals were divided by age groups: animals younger than 4 years (Age I), animals between 4 and 8 years of age (Age II), and animals between 8 and 15 years of age (Age III). In another analysis, the animals were grouped according to the sperm motility pattern obtained from the semen collection: sperm motility lower than 40% (Motility I), sperm motility between 40 and 70% (Motility II) and sperm motility between 70 and 90% (Motility III). The results of the present study demonstrated an effect of age (P < 0.05) on the following androgical characteristics: scrotal circumference, ejaculate volume, sperm vigor, major defects, minor defects and total defects. It was also observed that the animals with higher sperm motility presented higher (P < 0.05) scrotal circumference, and lower (P < 0.05) percentages of major and total defects. Among the andrological characteristics evaluated in the present work, it was observed positive correlations between age and scrotal circumference (R =0.299; P =0.000), age and volume of ejaculate (R =0.161; P =0.037), age and major defects (R =0.188; P =0.015), sperm motility and scrotal circumference (R =0.245; P =0.001), sperm motility and sperm vigor (R =0.483; P =0.000), and between major defects and total defects (R =0.946; P =0.000). Also, negative correlations were observed between sperm motility and total defects (R =-0.372; P =0.000), sperm vigor and major defects (R =-0.498; P =0.000), and sperm vigor and total defects (R =-0.432; P =0.000).Discussion: Based on the results of this study it was concluded that the Brangus-Ibage bulls utilized for natural breeding in eastern Mato Grosso do Sul, Brazil, presented satisfactory semen quality taking into account the quality of the pastures where the animals were located. In addition, the scrotal circumference, the ejaculate volume, the sperm vigor, and the percentage of morphological characteristics were influenced by the age of the bulls. Therefore, considering the production system and the environmental conditions, the animals with age between 4 and 8 years were superior regarding the sperm parameters evaluated.

Fertility Rates Following Fixed-Time Artificial Insemination in Dairy Heifers in a Practical Progesterone-Based Protocol

Background: In bovines, more efficient management practices are important for maximizing profitability. In order to increase the pregnancy rates in artificial insemination (AI) programs, several hormonal protocols were developed to synchronize the follicular wave and the moment of ovulation in beef and dairy cattle. In dairy cattle, detection of estrus can be difficult due to a number of factors including the incidence of silent estrus. Hormonal treatments designed to control both luteal and follicular function has permitting efficient synchronizations of time of ovulation. Thus, the AI can be performed in a large number of animals on a fixed schedule without the need for detection of estrus. Using these management techniques, the fixed-time artificial insemination (TAI) can overcome the problem of accurate estrus detection and help in reducing the incidence of repeat breeding. In addition, with TAI in cattle operations, it is possible to facilitate management practices and commercialization, and to reduce the time and semen wasting with animals inseminated at incorrect times. The investigation of practical and efficient TAI protocols is important for reducing the labor and animal handling of TAI in dairy cattle, as well as for increasing the profitability of the cattle management system. This study was carried out in order to investigate the effectiveness of TAI in dairy heifers treated with a practical progesterone-based protocol.Materials, Methods & Results: This experiment was conducted at the university farm located in southwestern Brazil, during May 2009. Thirty-nine cycling crossbred dairy heifers were employed in this study. All animals received a single intramuscular injection of estradiol benzoate and intravaginal progesterone releasing device in a random stage of the estrous cycle (Day 0). on day 7 the animals were treated with PGF2a analogue and on day 9 the device was removed. Forty-eight hours after the device removal (day 11) a synthetic analogue of GnRH was administered and the animals were fixed-time artificially inseminated at the time of GnRH injection. The inseminations were performed using four different batches from the same Holstein bull. Among the heifers that were synchronized (87.2%), 30.8% ovulated until 24 h after TAI and 56.4% ovulated between 24 and 32 h after TAI. The conception rate was 61.5%. No effects of ovulation time in conception rates were detected. The conception rate from heifers that ovulated until 24 h after TAI was 58.3% and from heifers that ovulated between 24 and 32 h after TAI was 77.3%. The mean of ovulatory follicle in heifers that ovulated until 24 h was 14.3 mm and in heifers that ovulated between 24 and 32 h was 11.9 mm.Discussion: Taking together, the findings of the present study, along with those of others, emphasize the concept that development of practical methods for TAI offers significant advantages to dairy producers if conception rates are close or greater to those obtained after breeding at detected estrus. Thus, the results of the present study reinforce the possibility of making dairy cattle production more cost-effective using TAI. In conclusion, with the progesterone-based TAI protocol of the present experiment all synchronized animals ovulated up to 32 h after GnRH+TAI and no effects of ovulation time related to conception rate was detected. The exogenous control of luteal and follicular development facilitated the reproductive management and animal handling. Also, inseminating the heifers at the moment of GnRH injection in a progesterone-based TAI protocol is a practical strategy and provided satisfactory results regarding ovulation and conception rates in dairy heifers.

Transmission electron microscopy for characterization of acrosomal damage after Percoll gradient centrifugation of cryopreserved bovine spermatozoa

The objective of this study was to characterize acrosomal ultrastructure following discontinuous Percoll gradient centrifugation of cryopreserved bovine sperm. Semen was collected from six bulls of different breeds and three ejaculates per bull were evaluated. Frozen semen samples were thawed and the acrosomal region of sperm cells was evaluated by transmission electron microscopy (TEM) before (n = 18) and after (n = 18) Percoll centrifugation. The evaluation of 20 sperm heads from each of the 36 samples analyzed ensured that a large number of cells were investigated. The data were subjected to analysis of variance at a level of significance of 5%. Percoll centrifugation reduced the percentage of sperm exhibiting normal acrosomes (from 61.77 to 30.24%), reduced the percentage of sperm presenting atypical acrosome reactions (from 28.38 to 4.84%) and increased the percentage of sperm exhibiting damage in the acrosome (from 6.14 to 64.26%). The percentage of sperm with typical acrosome reactions was not significantly different before (3.70%) and after (0.67%) centrifugation. TEM distinguished four different types of acrosomal status and enabled ultrastructural characterization of acrosomal injuries. The percentage of sperm exhibiting normal acrosomes decreased and damage in the acrosome was the most frequent acrosomal injury with the Percoll gradient centrifugation protocol utilized.

Assessment of field fertility and several in vitro sperm characteristics following the use of different Angus sires in a timed-AI program with suckled Nelore cows

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Suckling and allosuckling in river buffalo calves and its relation with weight gain

The aim of the present study was to investigate if river buffalo calves (Bubalus bubalis) have equal access to all cows and if milk is thus equally available to all of them. We recorded suckling/allosuckling behaviour and weight gain (WG) of 29 calves (14 males and 15 females), with special consideration to their sex, birth order (BO) and age. Cows' nursing behaviour and milli production (MP) were also considered. While males tended to be born earlier than females during this study, this was not the trend in the overall herd records. The cows' MP was not effected by the calves' sex. However, bull-calves presented greater mean WG, and mean times spent in individual filial (IF) and in communal nonfilial (CNF) suckling than heifer-calves, which showed greater communal filial (CF) suckling than the former during the first 4 months of life. The WG was associated with IF for bull-calves (r = 0.680 and 0.765, respectively, for the periods from birth to 4th and 8th months of age), and to CNF for heifer-calves (r = 0.628, for the period from birth to 8th month). Results from multiple regression analysis showed independent effects of each suckling category on the calf WG, and such effects were variable according to the calf's sex. BO was negatively correlated to calves' WG (bull-calves: r(s) = - 0.873 and - 0.799, from birth to 4th and gth months, respectively; heifer-calves: r(s) = - 0.531 from birth to 4th month). Specifically for bull-calves, there was a positive correlation between BO and MP (r(s) = 0.528 and 0.633, from birth to 4th and 8th months of age, respectively). The correlation between BO and IF was negative in both sexes, indicating that calves that were born early had more opportunities to suckle individually from their mothers. For heifer-calves, BO was positively correlated with CF (two periods), and negatively with CNF (from birth to 8th month of age), suggesting that heifer-calves were most often accompanied by other calves during suckling when they were born later. The data taken together indicate that sex and/or BO influenced decisively social interactions during suckling, promoting differential development among the calves. In animal husbandry, if a homogenous WG is desired, these factors have to be taken into consideration. (C) 2000 Elsevier B.V. B.V. All rights reserved.

Efeito da Suplementação Protéica sobre Crescimento, Terminação e Viabilidade Econômica de Bezerros Mestiços Canchim Confinados Pós-desmama

The feedlot performance of 30 post-weaning 3/4 Canchim 1/4 Nellore calves, with seven months of age and 208,8 kg of liveweight, was evaluated. Three different protein sources in isocaloric and isonitrogenous diets with 60:40 forage: concentrate ratio were used. The diets, fed ad libitum, constituted of corn silage, corn grain, mineral mix and a different protein source for each treatment: cottonseed meal, soybean meal and whole soybean. The weight gain, feed: gain ratio, dry matter and crude protein intake were evaluated for the growing (84 days) and fattening phases (84 days) and for the overall period. An economic study of young bulls production model was realized at the end of experimental period. Soybean meal based diet presented the best animal performance and provided R$ 51.30/head of net profit - the best economic result.

Seleção para peso pós-desmama em um rebanho Gir. 1. Resposta direta e correlacionada em 21 anos de seleção

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