Sexagem de espermatozoides bovinos por centrifugação em gradiente descontínuo de densidade de Percoll

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Qualidade da carne ovina submetida ao processo de salga

Foi estudada a qualidade da carne ovina submetida a diferentes teores de sal no processo de salga, utilizando-se 18 pernas desossadas provenientes de cordeiros ½ Ile de France ½ Ideal, machos inteiros, com peso vivo médio de 30kg. Os tratamentos constituíram-se em: T1 - controle; T2 - salga a 15% e T3 - salga a 20%. As mantas de carne foram lavadas rapidamente em água corrente para remoção do excesso de sal e secadas à sombra. As análises foram realizadas antes da salga e 67 horas após o início da mesma. As perdas de peso ao cozimento, maciez, cor, umidade (2, 4, 16, 28, 40 e 67 horas após o início da salga) e número de TBARS, substâncias reativas ao ácido 2-tiobarbitúrico, foram realizadas no músculo Semimembranosus. Os diferentes teores de sal não influenciaram (P>0,05) na luminosidade da carne, entretanto os teores de vermelho e amarelo, que foram influenciados (P<0,05). As perdas de peso ao cozimento e a umidade foram menores (P<0,05) nas carnes salgadas. A carne dos tratamentos controle e com 15% de salga, apresentou maior (P<0,05) maciez, em relação à carne do tratamento com 20% de salga. A carne que foi submetida a 20% de salga, apresentou maior oxidação lípidica, não diferindo da salga a 15%. A salga a 15% forneceu melhores resultados com valores de perdas de peso ao cozimento, cor, umidade e número de TBARS semelhante a salga a 20%, porém com maior maciez.

Características físico-químicas e sensoriais da carne de cordeiros terminados com dietas contendo cana-de-açúcar ou silagem de milho e dois níveis de concentrado

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Estudo preliminar da influência do perímetro escrotal sobre a libido em touros jovens da raça Nelore

O perímetro escrotal (PE) de 202 touros jovens da raça Nelore, sem experiência sexual prévia, classificados pela Associação Brasileira de Criadores de Zebu como superiores ou elite, foi medido aos 7, 12, 18 e 28 meses de idade, aproximadamente. Após a última medição, eles foram submetidos ao teste de libido e procedeu-se à colheita de sêmen por eletroejaculação, seguida de avaliação andrológica. As medidas do PE aos 7, 12, 18 e 28 meses foram de 18,30; 22,29; 27,54 e 33,26cm, respectivamente. Os coeficientes de correlação simples entre estas medidas variaram de 0,35 a 0,53. O coeficiente de correlação entre PE aos 28 meses e peso corporal na mesma idade foi de 0,43. A correlação entre a nota da libido e o perímetro escrotal medido aos l8 meses foi de 0,15 e entre aquela e o perímetro escrotal aos 28 meses foi de 0,13. Observou-se elevado crescimento do perímetro escrotal entre o 7º e 18º meses de idade. Utilizou-se o método de quadrados mínimos para analisar a libido, incluindo ano de nascimento como efeito fixo e como covariáveis os efeitos lineares da idade e peso no momento da avaliação e medidas de perímetro escrotal aos 7, l2, l8 e 28 meses. Cada covariável foi retida de forma seqüencial obtendo-se vários modelos de análises e as somas de quadrados foram decompostas de forma seqüencial e parcial. O ano de nascimento, a idade e o perímetro escrotal aos 18 meses de idade afetaram significativamente a libido. Os coeficientes de correlação entre a libido e concentração espermática, motilidade, peso e idade do animal no momento da avaliação foram de 0,34, 0,l6, 0,38 e 0,35, respectivamente.

Avaliação de dois métodos para condicionamento e coleta de sêmen em quatro espécies do gênero Mazama

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Produção in vitro de embriões bovinos: utilização de diferentes fontes de gonadotrofinas na maturação dos oócitos

O presente trabalho foi conduzido com o objetivo de avaliar o efeito da utilização de diferentes fontes de gonadotrofinas para maturação in vitro dos oócitos bovinos fecundados e desenvolvidos in vitro sobre as taxas de clivagem (TC) e de blastocistos (TBL). Oócitos imaturos provenientes de ovários de vacas de abatedouro foram submetidos a maturação in vitro sob diferentes condições: meio TCM 199, acrescido de 10% de soro de vaca em estro (SVE), aditivos, hepes, NaHCO3, piruvato de sódio, antibióticos (meio B-199), 20 UI/mL de PMSG e 10 UI/mL de hCG (PMSG/hCG) ou meio B-199, acrescido de 5 mig/mL de FSH e 5 mig/mL de LH (FSH/LH). Seguidos 24 h de cultura a 38,5ºC em atmosfera com 5% de CO2, os oócitos maturos foram incubados com sêmen descongelado durante 18 a 21 horas. Após esse período, os oócitos foram transferidos para placas contendo microgotas de meio Ménezo suplementado com 10% de SVE e células epiteliais do oviduto bovino em suspensão, cobertas com óleo de silicone, os quais permaneceram em cultura por mais 9 dias. Os dados foram analisados pelo teste do Qui-quadrado. A TC e a TBL, para PMSG/hCG e FSH/LH, foram 60 e 13,9% e 61,2 e 10,6%, respectivamente. Não houve diferença entre os tratamentos com relação a TC ou a TBL. Esses resultados sugerem que ambas as fontes de gonadotrofinas podem ser utilizadas para maturação in vitro dos oócitos fecundados e desenvolvidos in vitro.

Uso combinado de sêmen sexado e acasalamento dirigido sobre uma população de bovinos de corte submetida a seleção: estudo de simulação

Desenvolveu-se um estudo de simulação estocástica com o objetivo de verificar as consequências do uso combinado de acasalamento dirigido e sêmen sexado em uma população de bovinos de corte sob seleção. Simularam-se seis gerações de seleção para três cenários de acasalamento e uso de sêmen sexado. O primeiro cenário foi caracterizado por acasalamento aleatório e uso exclusivo de sêmen convencional. O segundo cenário caracterizou-se pelo uso de acasalamento associativo positivo nas 40% melhores vacas e acasalamento associativo negativo nas demais, sem uso de sêmen sexado. O terceiro cenário seguiu o mesmo procedimento de acasalamento do segundo, combinando-o com uso de sêmen sexado nas vacas submetidas a acasalamento associativo positivo. O acasalamento associativo positivo teve maior impacto no progresso genético que o uso de sêmen sexado, apesar de ter aumentado a incidência de endogamia na população. O uso de acasalamento associativo negativo foi ineficiente em reduzir a variabilidade dos animais destinados ao abate. O uso combinado de acasalmento associativo positivo e sêmen sexado aumentou a produção de animais geneticamente superiores.

Effects of scrotal insulation in Nellore bulls (Bos taurus indicus) on seminal quality and its relationship with in vitro fertilizing ability

The objectives of this study were to assess the effects of induced testicular degeneration in Bos taurus indicus (Nellore) bulls on changes in seminal characteristics and fertilizing ability of sperm. Four Nellore bulls (30-36-month-old, 500-550 kg) with good seminal quality (> 80% motile and morphologically normal sperm) had serotal insulation applied for 5 d. Semen was collected by electroejaculation and cryopreserved at a the pre-insulation moment, and 7, 14, and 21 d after insulation was removed. Gross motility, vigor of sperm movement (1-5), acrosome integrity, sperm morphology (phase-contrast microscopy), nuclear vacuoles and abnormal chromatin (Feulgen-stain) were determined after sperm preparations for in vitro fertilization (IVF). Prior to IVF, sperm were separated using a Percoll gradient (45% and 90%). Normal sperm decreased (P < 0.05) 14 and 21 d after insulation was removed. on 14 and 21 d, the incidence of head defects (9.7 +/- 0.6 and 17.0 +/- 0.8, respectively; mean +/- S.E.M) was higher (P < 0.05) in agreement with the incidence of nuclear vauoles (14.0 +/- 5.0 and 12.3 +/- 2.3) and abnormal chromatin (24.4 +/- 7.2 and 30.8 +/- 2.8). Although the frequency of cleaved oocytes decreased only on 21 d (P < 0.05), blastocyst rates were lower (P < 0.05) than pre-insulation on 14 and 21 d. In regression analyses, only nuclear vacuoles, head defects and intact acrosome accounted for differences in cleavage (R(2) = 0.38, 0.48, and 0.30, respectively) and blastocyst rates (R(2) = 0.35, 0.37, and 0.44). Abnormal chromatin was associated only with blastocyst rates (R(2) = 0.35). In conclusion, blastocyst rate was more sensitive than cleavage rate and the assessment of nuclear integrity is recommended to predict the fertilizing ability of bull sperm. (c) 2008 Elsevier B.V. All rights reserved.

Structural analysis of fertilization in the fish Brycon orbignyanus

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estrutura do dossel e acúmulo de forragem dos capins piatã e marandu sob pastejo com ovinos

The objective of this scientific work was to evaluate the tillering and mass production of Brachiaria brizantha Piatã and Marandu grasses with two statures of post-grazing. The experimental lineation was in factorial arrangement (2 pastures x 2 residue stature) with subplots (grazing cycle). The experimental period was from March to October of 2012. The target stature on pre-grazing was 50 cm and on post-grazing 15 and 25 for each cultivar. The following variables were analyzed: tiller population density, forage mass, morphological components, accumulation and accumulation rate. Three grazing cycles were obtained. Marandu grass presented larger forage mass on pre-grazing and lower stem production than piatã grass. No difference was observed between the two cultivars regarding the leaf blade production. There was no difference on forage mass, leaf blade, stem, and dead material production between the two statures of post-grazing. The pastures managed at 25 cm presented accumulation rate compared to those with 15 cm. Concerning the accumulation rate on the two cultivars, piatã and marandu grasses, Marandu grass presented higher amount than the piatã grass, and also presented higher values on the rate of basal tiller appearing on 15 cm stature post-pasture feature. Piatã grass managed at 25 cm developed smaller tiller amount on the second generation and greater tiller amount on the third generation than marandu grass. The greater tiller population density was observed on 25cm post-grazing pastures. There was no statistical difference of density between the two cultivars and between the cycles. Accordingly, it is concluded that piatã and marandu cultivars can be managed with 15cm and 25 cm post-grazing statures

Spermatozoon ultrastructure and sperm cryopreservation of the Brazilian dry season spawner fish pirapitinga, Brycon nattereri

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)