Effect of sheep diets with macadamia by-product and protected fat on the quality of fresh and frozen semen / Efeito de dietas de ovinos com coproduto de macadâmia e gordura protegida na qualidade de sêmen fresco e congelado

The ruminant diet is characterized by low lipid concentration, resulting from traditional diets composed by forage species. The use of agro industrial byproducts in animal feed may be interesting, once it reduces production costs and reduces environmental contamination. Among them, macadamia is known for interesting protein and carbohydrate contents; however, it is the amount of lipids that make it different. Fat supplementation can raise concentrations of blood cholesterol, a precursor metabolite of steroid hormones, which constitute biological membranes and possess specific and essential biological activities. The semen characteristics should be taken into account in the selection of the breeding herds, and the semen analysis makes it possible to evaluate the fertility of the sheep and allows obtaining important conclusions based on its results. The objective was to evaluate the seminal quality of Morada Nova sheep breed consuming diets supplemented with macadamia residue and protected fat. The experiment was carried out with 24 rams aged 18 or 30 months, distributed in four treatment groups: control (C), 50 g (MAC50) or 150 g (MAC150) of macadamia industrial byproduct; and 50 g of protected fat (Megalac®), added to the concentrate. Semen was collected at four intervals: before supplementation (day 0), 30, 60 and 75 days after the beginning of supplementation, and it was taken the measurements of volume, appearance, motility, vigour, turbulence, concentration and morphology. At days 60 and 75, semen was frozen for determination of plasma membrane integrity, acrosome integrity and mitochondrial activity after thawing. Analysis of variance was performed and the means were compared by the SNK test. In the analysis of fresh semen, a significant effect (p0.05). The inclusion of 50 or 150 g of macadamia residue or 50 g of Megalac in the...

A dieta dos ruminantes é caracterizada por baixa concentração de lipídeos, resultante de dietas tradicionais compostas por espécies forrageiras. A utilização de coprodutos agroindustriais na alimentação animal pode ser interessante, pois além de reduzir custos na produção, reduz a contaminação ambiental. Dentre eles a macadâmia é conhecida por teores interessantes de proteína e de carboidratos; entretanto é a quantidade de lipídeos que a torna diferenciada. A suplementação de gordura pode elevar as concentrações de colesterol sanguíneo, metabólito precursor dos hormônios esteroides, que constituem membranas biológicas e possuem atividades biológicas específicas e essenciais. As características seminais devem ser levadas em consideração na seleção dos reprodutores, sendo que a análise do sêmen possibilita avaliar a fertilidade do carneiro e permite obter importantes conclusões a partir dos seus resultados. O objetivo foi avaliar a qualidade seminal de carneiros Morada Nova consumindo dietas suplementadas com resíduo de macadâmia e gordura protegida. O experimento foi conduzido com 24 carneiros, com idade entre 18 e 30 meses, distribuídos em quatro grupos de tratamento: controle (C), 50 g (MAC50) ou 150 g (MAC150) de subproduto industrial da macadâmia; e 50 g de gordura protegida (Megalac®), adicionados ao concentrado. O sêmen foi coletado em quatro intervalos: antes da suplementação (dia 0), 30, 60 e 75 dias após o início da suplementação. O sêmen foi coletado para avaliação do volume, aparência, motilidade, vigor, turbilhonamento, concentração e morfologia. Nos dias 60 e 75, o sêmen foi congelado para determinação da integridade da membrana plasmática, integridade do acrossoma e atividade mitocondrial após o descongelamento. A análise de variância foi realizada e as médias comparadas pelo teste SNK. Na análise do sêmen fresco, foi observado efeito significativo (p<0,05) dos tratamentos na motilidade. Para...

Effect of sheep diets with macadamia by-product and protected fat on the quality of fresh and frozen semen / Efeito de dietas de ovinos com coproduto de macadâmia e gordura protegida na qualidade de sêmen fresco e congelado

The ruminant diet is characterized by low lipid concentration, resulting from traditional diets composed by forage species. The use of agro industrial byproducts in animal feed may be interesting, once it reduces production costs and reduces environmental contamination. Among them, macadamia is known for interesting protein and carbohydrate contents; however, it is the amount of lipids that make it different. Fat supplementation can raise concentrations of blood cholesterol, a precursor metabolite of steroid hormones, which constitute biological membranes and possess specific and essential biological activities. The semen characteristics should be taken into account in the selection of the breeding herds, and the semen analysis makes it possible to evaluate the fertility of the sheep and allows obtaining important conclusions based on its results. The objective was to evaluate the seminal quality of Morada Nova sheep breed consuming diets supplemented with macadamia residue and protected fat. The experiment was carried out with 24 rams aged 18 or 30 months, distributed in four treatment groups: control (C), 50 g (MAC50) or 150 g (MAC150) of macadamia industrial byproduct; and 50 g of protected fat (Megalac®), added to the concentrate. Semen was collected at four intervals: before supplementation (day 0), 30, 60 and 75 days after the beginning of supplementation, and it was taken the measurements of volume, appearance, motility, vigour, turbulence, concentration and morphology. At days 60 and 75, semen was frozen for determination of plasma membrane integrity, acrosome integrity and mitochondrial activity after thawing. Analysis of variance was performed and the means were compared by the SNK test. In the analysis of fresh semen, a significant effect (p<0.05) of the treatments on motility was observed. For cryopreserved semen, there was no significate difference (p>0.05). The inclusion of 50 or 150 g of macadamia residue or 50 g of Megalac in the...(AU)

A dieta dos ruminantes é caracterizada por baixa concentração de lipídeos, resultante de dietas tradicionais compostas por espécies forrageiras. A utilização de coprodutos agroindustriais na alimentação animal pode ser interessante, pois além de reduzir custos na produção, reduz a contaminação ambiental. Dentre eles a macadâmia é conhecida por teores interessantes de proteína e de carboidratos; entretanto é a quantidade de lipídeos que a torna diferenciada. A suplementação de gordura pode elevar as concentrações de colesterol sanguíneo, metabólito precursor dos hormônios esteroides, que constituem membranas biológicas e possuem atividades biológicas específicas e essenciais. As características seminais devem ser levadas em consideração na seleção dos reprodutores, sendo que a análise do sêmen possibilita avaliar a fertilidade do carneiro e permite obter importantes conclusões a partir dos seus resultados. O objetivo foi avaliar a qualidade seminal de carneiros Morada Nova consumindo dietas suplementadas com resíduo de macadâmia e gordura protegida. O experimento foi conduzido com 24 carneiros, com idade entre 18 e 30 meses, distribuídos em quatro grupos de tratamento: controle (C), 50 g (MAC50) ou 150 g (MAC150) de subproduto industrial da macadâmia; e 50 g de gordura protegida (Megalac®), adicionados ao concentrado. O sêmen foi coletado em quatro intervalos: antes da suplementação (dia 0), 30, 60 e 75 dias após o início da suplementação. O sêmen foi coletado para avaliação do volume, aparência, motilidade, vigor, turbilhonamento, concentração e morfologia. Nos dias 60 e 75, o sêmen foi congelado para determinação da integridade da membrana plasmática, integridade do acrossoma e atividade mitocondrial após o descongelamento. A análise de variância foi realizada e as médias comparadas pelo teste SNK. Na análise do sêmen fresco, foi observado efeito significativo (p<0,05) dos tratamentos na motilidade. Para...(AU)

The use of reduced glutathione (GSH) as antioxidant for cryopreserved sperm in dogs / O uso da glutationa reduzida (GSH) como antioxidante na criopreservação seminal em cães

The aim of this study was to evaluate the effect of supplementation with different concentrations of reduced glutathione GSH (0; 5; 7.5; 10mM) in the extender for cryopreservation in dogs with evaluations performed after glycerolization (chilled) and thawing (thawed). For this purpose, we used 8 dogs and two semen collections were performed in a weekly interval, totaling 16 semen samples. The sperm were analyzed by automatic sperm motility (CASA) and flow cytometry analysis of mitochondrial potential (JC1 dye) and membrane/acrosome integrity (FITC-PI dyes). We evaluated subjectively the membrane and acrosome integrity, mitochondrial activity and DNA integrity. Seminal plasma was evaluated for lipid peroxidation (TBARS concentration). Chilled and thawed samples supplemented with 7.5 and 10mM of GSH had lower percentage of sperm with high (DAB - Class I) and medium (DAB - Class II) mitochondrial activity. And 10mM of GSH had higher percentage of low mitochondrial activity (DAB - Class III). Moreover, thawed samples of 10mM of GSH had high DNA fragmentation rates. Probably by a reductive stress effect on mitochondria which lead to an increase in reactive oxygen species, and a mitochondrial malfunction.(AU)

O objetivo deste estudo foi avaliar o efeito da suplementação com diferentes concentrações de glutationa reduzida (GSH - 0; 5; 7,5; 10mM) para criopreservação em cães com avaliações realizadas após glicerolização (refrigeração) e descongelação. Para tal, foram utilizados oito cães e foram realizadas duas coletas de sêmen em intervalo semanal, totalizando 16 amostras de sêmen. Foram avaliadas a motilidade espermática computadorizada (CASA) e a análise de citometria de fluxo do potencial mitocondrial (sonda JC-1) e integridade da membrana/acrossomal (sonda FITC-PI). Subjetivamente foi avaliada a integridade da membrana plasmática e do acrossomal, atividade mitocondrial e integridade do DNA. O plasma seminal foi avaliado quanto à peroxidação lipídica (concentração de TBARS). As amostras refrigeradas e descongeladas suplementadas com 7,5 e 10mM de GSH apresentaram menor porcentagem de espermatozoides com alta atividade mitocondrial (DAB - Classe I) e média (DAB - Classe II). Na concentração de 10mM de GSH, apresentaram maior porcentagem de baixa atividade mitocondrial (DAB - Classe III). Além disso, amostras descongeladas de 10mM de GSH apresentaram taxas de fragmentação de DNA elevadas, provavelmente por efeito de estresse redutivo sobre as mitocôndrias que elevam as espécies reativas de oxigênio e disfunção mitocondrial.(AU)

The use of reduced glutathione (GSH) as antioxidant for cryopreserved sperm in dogs / O uso da glutationa reduzida (GSH) como antioxidante na criopreservação seminal em cães

The aim of this study was to evaluate the effect of supplementation with different concentrations of reduced glutathione GSH (0; 5; 7.5; 10mM) in the extender for cryopreservation in dogs with evaluations performed after glycerolization (chilled) and thawing (thawed). For this purpose, we used 8 dogs and two semen collections were performed in a weekly interval, totaling 16 semen samples. The sperm were analyzed by automatic sperm motility (CASA) and flow cytometry analysis of mitochondrial potential (JC1 dye) and membrane/acrosome integrity (FITC-PI dyes). We evaluated subjectively the membrane and acrosome integrity, mitochondrial activity and DNA integrity. Seminal plasma was evaluated for lipid peroxidation (TBARS concentration). Chilled and thawed samples supplemented with 7.5 and 10mM of GSH had lower percentage of sperm with high (DAB - Class I) and medium (DAB - Class II) mitochondrial activity. And 10mM of GSH had higher percentage of low mitochondrial activity (DAB - Class III). Moreover, thawed samples of 10mM of GSH had high DNA fragmentation rates. Probably by a reductive stress effect on mitochondria which lead to an increase in reactive oxygen species, and a mitochondrial malfunction.(AU)

O objetivo deste estudo foi avaliar o efeito da suplementação com diferentes concentrações de glutationa reduzida (GSH - 0; 5; 7,5; 10mM) para criopreservação em cães com avaliações realizadas após glicerolização (refrigeração) e descongelação. Para tal, foram utilizados oito cães e foram realizadas duas coletas de sêmen em intervalo semanal, totalizando 16 amostras de sêmen. Foram avaliadas a motilidade espermática computadorizada (CASA) e a análise de citometria de fluxo do potencial mitocondrial (sonda JC-1) e integridade da membrana/acrossomal (sonda FITC-PI). Subjetivamente foi avaliada a integridade da membrana plasmática e do acrossomal, atividade mitocondrial e integridade do DNA. O plasma seminal foi avaliado quanto à peroxidação lipídica (concentração de TBARS). As amostras refrigeradas e descongeladas suplementadas com 7,5 e 10mM de GSH apresentaram menor porcentagem de espermatozoides com alta atividade mitocondrial (DAB - Classe I) e média (DAB - Classe II). Na concentração de 10mM de GSH, apresentaram maior porcentagem de baixa atividade mitocondrial (DAB - Classe III). Além disso, amostras descongeladas de 10mM de GSH apresentaram taxas de fragmentação de DNA elevadas, provavelmente por efeito de estresse redutivo sobre as mitocôndrias que elevam as espécies reativas de oxigênio e disfunção mitocondrial.(AU)

Effect of Prostaglandin F2 at the beginning of the ovulation synchronization protocol in dairy Bos indicus, Bos taurus and Bos indicus x Bos taurus heifers / Efeito da Prostaglandina F2 no início do protocolo de sincronização da ovulação em novilhas leiteiras Bos indicus, Bos taurus e Bos indicus x Bos taurus

The objective of the present study was to evaluate the effect of treatment with prosta-glandin F2α (PGF2α) at the beginning of the protocol for ovulation synchronization on follicular dy-namics in Bos indicus (Gyr; n=11), Bos taurus (Holstein Black and White, HBW; n=10), and crossbred animals (Gyr x HBW; n=12). On a random day of the estrous cycles (day 0, D0), the animals received 2.0 mg estradiol benzoate (EB) intramuscularly plus an intravaginal progesterone (P4) device, which was maintained for 8 days (day 8, D8). Half the heifers of each group received a dose of 25 mg PGF2αintramuscularly at the time of insertion of the intravaginal P4 device. When the intravaginal device was removed (D8), all animals received another dose of 25 mg PGF2α intramuscularly, followed by intramuscular injection of 1.0 mg EB 24 h later (day 9, D9). Ultrasonographic evaluations were per-formed at intervals of 24 hours from D0 to D8 and at intervals of 12 hours from removal of the P4 device to 96 hours thereafter. Samples were collected on days 0, 3, 6, 8, 10 and 22 for the measurement of P4. The mean maximum diameter of the dominant follicle (DF) was smaller (P=0.01) in Gyr heifers (10.0 ± 0.8 mm) than in Gyr x HBW (13.0 ± 0.6 mm) or HBW (12.5 ± 0.8 mm). Furthermore, treatment with PGF2α on D0 increased (P=0.02) the maximum diameter of DF (12.9 ± 0.5 vs. 10.9 ± 0.7 mm)...

O objetivo do presente estudo foi avaliar o efeito do tratamento com prostaglandina F2α (PGF2α)no início do protocolo de sincronização da ovulação, sobre a dinâmica folicular de novilhas Bos indicus (Gir; n=11), Bos taurus (Holandesa Preto e Branco - HPB; n=10) e cruzadas (Gir x HPB; n=12). Em dia aleatório do ciclo estral (dia 0 - D0), os animais receberam 2,0 mg de benzoato de es-tradiol (BE) via intramuscular, mais um dispositivo intravaginal de progesterona (P4) que foi man-tido por oito dias (dia 8 – D8). Metade das novilhas de cada grupo recebeu uma dose de 25 mg de PGF2α via intramuscular no momento da inserção do dispositivo intravaginal de P4. Na retirada do dispositivo intravaginal (D8), todos os animais receberam outra dose de 25 mg de PGF2α via intra-muscular e 24 h após (dia 9 - D9) receberam 1,0 mg de BE via intramuscular. Foram realizadas ava-liações ultrassonográficas com intervalos de 24 horas do D0 até o D8 e com intervalos de 12 horas da retirada do dispositivo de P4 até 96 horas. Para a dosagem de P4, foram colhidas amostras nos dias 0; 3; 6; 8; 10 e 22. A média do diâmetro máximo do folículo dominante (FD) foi menor (P=0,01) nas novilhas Gir (10,0 ± 0,8 mm) do que nas cruzadas Gir x HPB (13,0 ± 0,6 mm) ou HPB (12,5 ± 0,8 mm). Além disso, o tratamento com PGF2α no D0 aumentou (P=0,02) o diâmetro máximo do FD (12,9 ± 0,5 mm vs. 10,9 ± 0,7 mm) nos três grupamentos genéticos avaliados. A taxa de crescimento do FD foi menor (P=0,008) nas novilhas Gir (0,8 ± 0,1 mm/dia), do que nas novilhas cruzadas (1,3 ± 0,1 mm/dia) ou HPB (1,2 ± 0,1 mm/dia)...

Effect of Prostaglandin F2 at the beginning of the ovulation synchronization protocol in dairy Bos indicus, Bos taurus and Bos indicus x Bos taurus heifers / Efeito da Prostaglandina F2 no início do protocolo de sincronização da ovulação em novilhas leiteiras Bos indicus, Bos taurus e Bos indicus x Bos taurus

The objective of the present study was to evaluate the effect of treatment with prosta-glandin F2α (PGF2α) at the beginning of the protocol for ovulation synchronization on follicular dy-namics in Bos indicus (Gyr; n=11), Bos taurus (Holstein Black and White, HBW; n=10), and crossbred animals (Gyr x HBW; n=12). On a random day of the estrous cycles (day 0, D0), the animals received 2.0 mg estradiol benzoate (EB) intramuscularly plus an intravaginal progesterone (P4) device, which was maintained for 8 days (day 8, D8). Half the heifers of each group received a dose of 25 mg PGF2αintramuscularly at the time of insertion of the intravaginal P4 device. When the intravaginal device was removed (D8), all animals received another dose of 25 mg PGF2α intramuscularly, followed by intramuscular injection of 1.0 mg EB 24 h later (day 9, D9). Ultrasonographic evaluations were per-formed at intervals of 24 hours from D0 to D8 and at intervals of 12 hours from removal of the P4 device to 96 hours thereafter. Samples were collected on days 0, 3, 6, 8, 10 and 22 for the measurement of P4. The mean maximum diameter of the dominant follicle (DF) was smaller (P=0.01) in Gyr heifers (10.0 ± 0.8 mm) than in Gyr x HBW (13.0 ± 0.6 mm) or HBW (12.5 ± 0.8 mm). Furthermore, treatment with PGF2α on D0 increased (P=0.02) the maximum diameter of DF (12.9 ± 0.5 vs. 10.9 ± 0.7 mm)...(AU)

O objetivo do presente estudo foi avaliar o efeito do tratamento com prostaglandina F2α (PGF2α)no início do protocolo de sincronização da ovulação, sobre a dinâmica folicular de novilhas Bos indicus (Gir; n=11), Bos taurus (Holandesa Preto e Branco - HPB; n=10) e cruzadas (Gir x HPB; n=12). Em dia aleatório do ciclo estral (dia 0 - D0), os animais receberam 2,0 mg de benzoato de es-tradiol (BE) via intramuscular, mais um dispositivo intravaginal de progesterona (P4) que foi man-tido por oito dias (dia 8 D8). Metade das novilhas de cada grupo recebeu uma dose de 25 mg de PGF2α via intramuscular no momento da inserção do dispositivo intravaginal de P4. Na retirada do dispositivo intravaginal (D8), todos os animais receberam outra dose de 25 mg de PGF2α via intra-muscular e 24 h após (dia 9 - D9) receberam 1,0 mg de BE via intramuscular. Foram realizadas ava-liações ultrassonográficas com intervalos de 24 horas do D0 até o D8 e com intervalos de 12 horas da retirada do dispositivo de P4 até 96 horas. Para a dosagem de P4, foram colhidas amostras nos dias 0; 3; 6; 8; 10 e 22. A média do diâmetro máximo do folículo dominante (FD) foi menor (P=0,01) nas novilhas Gir (10,0 ± 0,8 mm) do que nas cruzadas Gir x HPB (13,0 ± 0,6 mm) ou HPB (12,5 ± 0,8 mm). Além disso, o tratamento com PGF2α no D0 aumentou (P=0,02) o diâmetro máximo do FD (12,9 ± 0,5 mm vs. 10,9 ± 0,7 mm) nos três grupamentos genéticos avaliados. A taxa de crescimento do FD foi menor (P=0,008) nas novilhas Gir (0,8 ± 0,1 mm/dia), do que nas novilhas cruzadas (1,3 ± 0,1 mm/dia) ou HPB (1,2 ± 0,1 mm/dia)...(AU)

Factors affecting gestational length in the Mangalarga Paulista breed

The Mangalarga Paulista is a Brazilian horse breed that originated from a cross between Andalusian and Alter Real horses, and despite its national importance, no data exist regarding their gestational length. This study was conducted to determine the variation in gestation length and the influence of factors such as the sex of the foal, maternal age and parity, stallion and mating period. Retrospectively, data from 208 gestations of Mangalarga Paulista foals located in Piracaia, SP, Brazil, during the period from 1991 to 2012 were analyzed. The mean gestational length was 343.08 ± 0.66 days, with a range between 317 and 404 days. Gestation of mares carrying colts had a tendency to be longer than mares carrying fillies. Maternal parity significantly affected gestational length, with longer gestation for primiparous mares until the third gestation, shorter pregnancies between 4th and 8th gestation, and longer thereafter. Regarding mating period, mares that were mated at the end of the breeding season (January and February) had shorter pregnancies (P < 0.05). These results are of significant clinical and economic relevance for the equine industry, so that the monitoring of gestation and parturition can be performed effectively.

Factors affecting gestational length in the Mangalarga Paulista breed

The Mangalarga Paulista is a Brazilian horse breed that originated from a cross between Andalusian and Alter Real horses, and despite its national importance, no data exist regarding their gestational length. This study was conducted to determine the variation in gestation length and the influence of factors such as the sex of the foal, maternal age and parity, stallion and mating period. Retrospectively, data from 208 gestations of Mangalarga Paulista foals located in Piracaia, SP, Brazil, during the period from 1991 to 2012 were analyzed. The mean gestational length was 343.08 ± 0.66 days, with a range between 317 and 404 days. Gestation of mares carrying colts had a tendency to be longer than mares carrying fillies. Maternal parity significantly affected gestational length, with longer gestation for primiparous mares until the third gestation, shorter pregnancies between 4th and 8th gestation, and longer thereafter. Regarding mating period, mares that were mated at the end of the breeding season (January and February) had shorter pregnancies (P < 0.05). These results are of significant clinical and economic relevance for the equine industry, so that the monitoring of gestation and parturition can be performed effectively.(AU)

The influence of canine brucellosis on morphofunctional features of epididymal spermatozoa: case report / Influência da brucelose canina nos aspectos morfofuncionais de espermatozoides epididimários: relato de caso

The present work reports a clinical case of a mongrel dog, with serological diagnosis of brucellosis, from which epididymal sperm analysis was performed. Sperm samples were collected from different segments of the epididymis (tail, corpus, and caput). Sperm samples were evaluated for computer-assisted motility analysis (CASA), spermatic morphology, mitochondrial activity and sperm plasmatic membrane and acrosomal integrity. Changes in sperm movement patterns were found (progressive motility, percentage of rapid sperm, percentage of rapid velocity, average pathway, curvilinear velocity, velocity straight line, amplitude of lateral head displacement, straightness and linearity), increase of total morphological defects (51%) and absence of sperm mitochondrial activity (20%) were verified, especially for cauda epididymides. We highlight that such changes can contribute to clinical diagnosis of Brucellosis in dogs and to the use of epididymal sperm in reproductive biotechnologies.(AU)

Relata-se o caso de um cão mestiço, com diagnóstico sorológico para brucelose canina, a partir do qual foram realizadas análises do sêmen epididimário. As amostras espermáticas foram coletadas dos diferentes segmentos epididimários (cabeça, corpo e cauda). Foram realizadas as avaliações de motilidade computadorizada do sêmen (CASA), morfologia espermática, atividade mitocondrial, integridade das membranas plasmática e acrossomal. Houve alteração no padrão de movimentação espermática (motilidade progressiva, espermatozoides rápidos, velocidade média da trajetória, velocidade curvilínea, velocidade linear progressiva, amplitude de deslocamento lateral da cabeça, retilinearidade e linearidade), aumento do total de defeitos morfológicos (51%) e da ausência de atividade mitocondrial espermática (20%) dos espermatozoides, especialmente da cauda do epidídimo. Ressalta-se que tais achados podem contribuir para o diagnóstico clínico da brucelose canina e para a utilização do sêmen epididimário em biotecnologias da reprodução.(AU)

The influence of canine brucellosis on morphofunctional features of epididymal spermatozoa: case report / Influência da brucelose canina nos aspectos morfofuncionais de espermatozoides epididimários: relato de caso

The present work reports a clinical case of a mongrel dog, with serological diagnosis of brucellosis, from which epididymal sperm analysis was performed. Sperm samples were collected from different segments of the epididymis (tail, corpus, and caput). Sperm samples were evaluated for computer-assisted motility analysis (CASA), spermatic morphology, mitochondrial activity and sperm plasmatic membrane and acrosomal integrity. Changes in sperm movement patterns were found (progressive motility, percentage of rapid sperm, percentage of rapid velocity, average pathway, curvilinear velocity, velocity straight line, amplitude of lateral head displacement, straightness and linearity), increase of total morphological defects (51%) and absence of sperm mitochondrial activity (20%) were verified, especially for cauda epididymides. We highlight that such changes can contribute to clinical diagnosis of Brucellosis in dogs and to the use of epididymal sperm in reproductive biotechnologies.(AU)

Relata-se o caso de um cão mestiço, com diagnóstico sorológico para brucelose canina, a partir do qual foram realizadas análises do sêmen epididimário. As amostras espermáticas foram coletadas dos diferentes segmentos epididimários (cabeça, corpo e cauda). Foram realizadas as avaliações de motilidade computadorizada do sêmen (CASA), morfologia espermática, atividade mitocondrial, integridade das membranas plasmática e acrossomal. Houve alteração no padrão de movimentação espermática (motilidade progressiva, espermatozoides rápidos, velocidade média da trajetória, velocidade curvilínea, velocidade linear progressiva, amplitude de deslocamento lateral da cabeça, retilinearidade e linearidade), aumento do total de defeitos morfológicos (51%) e da ausência de atividade mitocondrial espermática (20%) dos espermatozoides, especialmente da cauda do epidídimo. Ressalta-se que tais achados podem contribuir para o diagnóstico clínico da brucelose canina e para a utilização do sêmen epididimário em biotecnologias da reprodução.(AU)

Utilização de pentoxifilina e antioxidantes na criopreservação do sêmen bovino: qualidade seminal e estresse oxidativo / Utilization of pentoxifylline and antioxidants in cryopreservation of bovine semen: semen quality and oxidative stress

O presente estudo avaliou a adição da pentoxifilina, tocoferol, ascorbato e suas combinações sobre a proteção da célula espermática bovina contra os efeitos deletérios da criopreservação. Foram utilizados 24 touros Nelores (Bos taurus indicus), criados em sistema semi-intensivo. Foi coletado um ejaculado de cada reprodutor, diluídos em TRIS-citratogema-glicerol e divididos em seis partes. Cada parte foi suplementada da seguinte maneira: sem aditivos (controle), tocoferol (10 mmol/mL), tocoferol (10 mmol/mL) + pentoxifilina (1mg/mL), ascorbato (0,45mg/mL), ascorbato (0,45mg/mL) + pentoxifilina (1mg/mL) ou pentoxifilina (1mg/mL). Após o descongelamento, as amostras foram avaliadas quanto à motilidade e características do movimento, integridade da membrana plasmática e de acrossomo e atividade mitocondrial. Os níveis de peroxidação lipídica espontânea e induzida foram avaliados pela produção de substâncias reativas ao ácido tiobarbitúrico (TBARS). A suplementação com pentoxifilina, tocoferol, ascorbato e suas combinações, não alterou (P>0,05) a atividade mitocondrial, integridade acrossomal, e a concentração de TBARS espontâneo e induzido. A adição de tocoferol + pentoxifilina reduziu a motilidade progressiva quando comparado ao ascorbato e também a integridade da membrana espermática quando comparado ao controle e ao ascorbato (P˂0,05). Já a adição de ascorbato + pentoxifilina foi deletéria sobre linearidade em comparação ao tratamento ascorbato (P˂0,05). A adição de ascorbato, tocoferol e pentoxifilina individualmente ou em combinação, não foi eficiente em diminuir os danos causados pela criopreservação e estresse oxidativo em amostras pós descongelamento de sêmen bovino.

The present study evaluated the addition of pentoxifylline, tocopherol and ascorbate and their combinations on the protection of bovine spermatic cells against the deleterious effects of cryopreservation. A total of 24 Nelore bulls (Bos Taurus indicus) were included in this study, raised in a semi-intensive system. One ejaculation was collected from each bull and diluted in tris-citrate-yolk-glycerol, divided into six portions, and then supplemented with: no additives (control); tocopherol (10 mmol/ml), tocopherol (10 mmol/ml) + pentxoifylline (1 mg/ml), ascorbate (0.45 mg/ml), ascorbate (0.45 mg/ml) + pentoxifylline (1 mg/ml) and pentxoifylline (1 mg/ml). After thawing, the samples were evaluated for motility and characteristics of movement, acrosomal and plasma membrane integrity, and mitochondrial activity. Spontaneous and induced lipid peroxidation levels were evaluated by the production of thiobarbituric acid reactive substances (TBARS). Supplementation with pentoxifylline, tocopherol and ascorbate and their combinations, did not alter mitochondrial activity, acrosomal integrity, or the spontaneous and induced TBARS concentration (P>0.05). The ascorbate, tocopherol and pentoxifylline additions, both individually and in combinations, were inefficient in decreasing the damage caused by cryopreservation and oxidative stress in post-thawed bovine semen samples.

Utilização de pentoxifilina e antioxidantes na criopreservação do sêmen bovino: qualidade seminal e estresse oxidativo / Utilization of pentoxifylline and antioxidants in cryopreservation of bovine semen: semen quality and oxidative stress

O presente estudo avaliou a adição da pentoxifilina, tocoferol, ascorbato e suas combinações sobre a proteção da célula espermática bovina contra os efeitos deletérios da criopreservação. Foram utilizados 24 touros Nelores (Bos taurus indicus), criados em sistema semi-intensivo. Foi coletado um ejaculado de cada reprodutor, diluídos em TRIS-citratogema-glicerol e divididos em seis partes. Cada parte foi suplementada da seguinte maneira: sem aditivos (controle), tocoferol (10 mmol/mL), tocoferol (10 mmol/mL) + pentoxifilina (1mg/mL), ascorbato (0,45mg/mL), ascorbato (0,45mg/mL) + pentoxifilina (1mg/mL) ou pentoxifilina (1mg/mL). Após o descongelamento, as amostras foram avaliadas quanto à motilidade e características do movimento, integridade da membrana plasmática e de acrossomo e atividade mitocondrial. Os níveis de peroxidação lipídica espontânea e induzida foram avaliados pela produção de substâncias reativas ao ácido tiobarbitúrico (TBARS). A suplementação com pentoxifilina, tocoferol, ascorbato e suas combinações, não alterou (P>0,05) a atividade mitocondrial, integridade acrossomal, e a concentração de TBARS espontâneo e induzido. A adição de tocoferol + pentoxifilina reduziu a motilidade progressiva quando comparado ao ascorbato e também a integridade da membrana espermática quando comparado ao controle e ao ascorbato (P˂0,05). Já a adição de ascorbato + pentoxifilina foi deletéria sobre linearidade em comparação ao tratamento ascorbato (P˂0,05). A adição de ascorbato, tocoferol e pentoxifilina individualmente ou em combinação, não foi eficiente em diminuir os danos causados pela criopreservação e estresse oxidativo em amostras pós descongelamento de sêmen bovino.(AU)

The present study evaluated the addition of pentoxifylline, tocopherol and ascorbate and their combinations on the protection of bovine spermatic cells against the deleterious effects of cryopreservation. A total of 24 Nelore bulls (Bos Taurus indicus) were included in this study, raised in a semi-intensive system. One ejaculation was collected from each bull and diluted in tris-citrate-yolk-glycerol, divided into six portions, and then supplemented with: no additives (control); tocopherol (10 mmol/ml), tocopherol (10 mmol/ml) + pentxoifylline (1 mg/ml), ascorbate (0.45 mg/ml), ascorbate (0.45 mg/ml) + pentoxifylline (1 mg/ml) and pentxoifylline (1 mg/ml). After thawing, the samples were evaluated for motility and characteristics of movement, acrosomal and plasma membrane integrity, and mitochondrial activity. Spontaneous and induced lipid peroxidation levels were evaluated by the production of thiobarbituric acid reactive substances (TBARS). Supplementation with pentoxifylline, tocopherol and ascorbate and their combinations, did not alter mitochondrial activity, acrosomal integrity, or the spontaneous and induced TBARS concentration (P>0.05). The ascorbate, tocopherol and pentoxifylline additions, both individually and in combinations, were inefficient in decreasing the damage caused by cryopreservation and oxidative stress in post-thawed bovine semen samples.(AU)

Evaluation of sperm quality in successive regular collections from captive black-and-gold howler monkeys (Alouatta caraya)

The black-and-gold howler monkey serves as a suitable model for development of assisted reproductive techniques (ARTs), which may later be applied in threatened species of the same genus. The objectives of this study were to evaluate semen characteristics of black-and-gold howler monkeys (Alouatta caraya), with emphasis on mitochondrial activity and acrosome integrity, using staining methods that are suitable for use under field conditions. Twenty six semen samples were collected by rectal probe electroejaculation from six adult captive males from the National Primate Center (CENP), Ananindeua, PA, Brazil. The following parameters were evaluated in each semen sample: volume, pH, concentration, plasma membrane integrity, acrosome integrity, and mitochondrial activity. Both Spermac stain and the Simple Staining Method were suitable for acrosome evaluation. Diaminobenzidine needs further investigation to improve its application for this species. Overall semen quality improved from the first collection to the fifth collection. This highlights the importance of repeating semen collections when evaluating a male for use in ARTs. This research brought novel information on semen characteristics of Alouatta caraya which will aid further studies on the application of ARTs in this and other primate species.

Evaluation of sperm quality in successive regular collections from captive black-and-gold howler monkeys (Alouatta caraya)

The black-and-gold howler monkey serves as a suitable model for development of assisted reproductive techniques (ARTs), which may later be applied in threatened species of the same genus. The objectives of this study were to evaluate semen characteristics of black-and-gold howler monkeys (Alouatta caraya), with emphasis on mitochondrial activity and acrosome integrity, using staining methods that are suitable for use under field conditions. Twenty six semen samples were collected by rectal probe electroejaculation from six adult captive males from the National Primate Center (CENP), Ananindeua, PA, Brazil. The following parameters were evaluated in each semen sample: volume, pH, concentration, plasma membrane integrity, acrosome integrity, and mitochondrial activity. Both Spermac stain and the Simple Staining Method were suitable for acrosome evaluation. Diaminobenzidine needs further investigation to improve its application for this species. Overall semen quality improved from the first collection to the fifth collection. This highlights the importance of repeating semen collections when evaluating a male for use in ARTs. This research brought novel information on semen characteristics of Alouatta caraya which will aid further studies on the application of ARTs in this and other primate species.(AU)

Sex-sorted sperm for artificial in semination and embryo transfer programs in cattle

The selection of offspring from the desired sex can be one of the determining factors to increase the genetic progress and farmer ́s profitability in either beef or dairy cattle. In fact, the use of sex-sorted sperm has been applied worldwide combined with artificial insemination (AI) upon estrus detection in heifers. Additionally, several researches have been performed aiming to investigate the use of sex-sorted sperm during timed AI (TAI) programs and for insemination of superstimulated donors for in vivo embryo programs. Pregnancy per AI (P/AI) of cyclic heifers inseminated in estrus with sex-sorted sperm has been reported to be approximately 75 to 80% of the P/AI of heifers inseminated with non-sorted sperm. Insemination of superstimulated cows with sex-sorted sperm has been reported to reduce the production of viable embryos. Recently, however, it has been demonstrated that P/AI and embryo production per flushing resulting from AI with sex-sorted sperm may be improved when the time of AI is postponed in relation to the time of AI with non-sorted sperm. The P/AI of non stimulated females and fertilization rates and number of embryos recovered from superstimulated females were increased when AI occurred between 16 and 24 h after the onset of estrus (i.e. 6 to 14 h before ovulation). Nonetheless, despite the improvements achieved in the last decade, there is still a significant individual variability in fertility among bulls that have their sperm se x-sorted. It is critical that the pre-determination of the sire fertility is a paramount when sex-sorted sperm is utilized in commercial AI and ET programs. Thus, the aim of this review is to discuss the main concepts related to the use of sex-sorted sperm in TAI and ET programs, addressing some strategies to increase the efficiency of the technique.

Sex-sorted sperm for artificial in semination and embryo transfer programs in cattle

The selection of offspring from the desired sex can be one of the determining factors to increase the genetic progress and farmer ́s profitability in either beef or dairy cattle. In fact, the use of sex-sorted sperm has been applied worldwide combined with artificial insemination (AI) upon estrus detection in heifers. Additionally, several researches have been performed aiming to investigate the use of sex-sorted sperm during timed AI (TAI) programs and for insemination of superstimulated donors for in vivo embryo programs. Pregnancy per AI (P/AI) of cyclic heifers inseminated in estrus with sex-sorted sperm has been reported to be approximately 75 to 80% of the P/AI of heifers inseminated with non-sorted sperm. Insemination of superstimulated cows with sex-sorted sperm has been reported to reduce the production of viable embryos. Recently, however, it has been demonstrated that P/AI and embryo production per flushing resulting from AI with sex-sorted sperm may be improved when the time of AI is postponed in relation to the time of AI with non-sorted sperm. The P/AI of non stimulated females and fertilization rates and number of embryos recovered from superstimulated females were increased when AI occurred between 16 and 24 h after the onset of estrus (i.e. 6 to 14 h before ovulation). Nonetheless, despite the improvements achieved in the last decade, there is still a significant individual variability in fertility among bulls that have their sperm se x-sorted. It is critical that the pre-determination of the sire fertility is a paramount when sex-sorted sperm is utilized in commercial AI and ET programs. Thus, the aim of this review is to discuss the main concepts related to the use of sex-sorted sperm in TAI and ET programs, addressing some strategies to increase the efficiency of the technique. (AU)

Reproductive characteristics of captive greater rhea (Rhea americana) males reared in the state of São Paulo, Brazil

Rheas (Rhea americana) belongs to the ratite group. Considering the commercial significance of this birds, some techniques, such as semen collection, were standardized. In this study, 107 male rheas (3 to 4 years of age) reared in commercial farms in the state of São Paulo, Brazil, were used. Semen was collected during the breeding and off-breeding seasons of 2001, 2002, and 2003. Bird hierarchical behavior was observed. Birds were restrained performed using a box and a black hood. Semen was collected by digital pressure on the base of the phallus, which size was measured, and the presence or absence of spiral shape was observed. Immediately after collection, semen samples were evaluated for volume, motility, sperm concentration, and morphology. In a limited number of birds, blood samples were collected to measure testosterone levels. Among the 69 birds studied during the breeding season, 44 presented large phalluses, out of which 26 showed spiral shape. The method of semen collection was efficient. The following semen parameter results were obtained: volume (0.68 ±0.14 ml), motility (61.11±11.54%), sperm concentration (3.29±1.33 x10(9) sptz/ml), and number of spermatozoa per ejaculate (2.40±1.38x10(9) sptz/ml). Morphological abnormalities were analyzed and recorded. Testosterone levels were statistically different (p = 0.0161) between the breeding and non-breeding season (53.28±18.41 ng/ml and 5:57±3.81 ng/ml, respectively). Variations in phallus size were also found between the breeding and non-breeding seasons. Larger phalluses and higher testosterone levels were correlated with dominant behavior. The results of the present experiment confirmed that it is possible to collect semen from rheas, allowing the future use of biotechnologies such as artificial insemination.

Reproductive characteristics of captive greater rhea (Rhea americana) males reared in the state of São Paulo, Brazil

Rheas (Rhea americana) belongs to the ratite group. Considering the commercial significance of this birds, some techniques, such as semen collection, were standardized. In this study, 107 male rheas (3 to 4 years of age) reared in commercial farms in the state of São Paulo, Brazil, were used. Semen was collected during the breeding and off-breeding seasons of 2001, 2002, and 2003. Bird hierarchical behavior was observed. Birds were restrained performed using a box and a black hood. Semen was collected by digital pressure on the base of the phallus, which size was measured, and the presence or absence of spiral shape was observed. Immediately after collection, semen samples were evaluated for volume, motility, sperm concentration, and morphology. In a limited number of birds, blood samples were collected to measure testosterone levels. Among the 69 birds studied during the breeding season, 44 presented large phalluses, out of which 26 showed spiral shape. The method of semen collection was efficient. The following semen parameter results were obtained: volume (0.68 ±0.14 ml), motility (61.11±11.54%), sperm concentration (3.29±1.33 x10(9) sptz/ml), and number of spermatozoa per ejaculate (2.40±1.38x10(9) sptz/ml). Morphological abnormalities were analyzed and recorded. Testosterone levels were statistically different (p = 0.0161) between the breeding and non-breeding season (53.28±18.41 ng/ml and 5:57±3.81 ng/ml, respectively). Variations in phallus size were also found between the breeding and non-breeding seasons. Larger phalluses and higher testosterone levels were correlated with dominant behavior. The results of the present experiment confirmed that it is possible to collect semen from rheas, allowing the future use of biotechnologies such as artificial insemination.

Follicular growth and plasma progesterone patterns in Bos indicus x Bos taurus heifers submitted to different PGF2/progesterone-based synchronization protocols

The effect of different levels of progesterone (P4) concentrations on follicle growth and ovulatory capacity was evaluated in 40 crossbred Bos indicus x Bos taurus cyclic heifers submitted to distinct PGF2α + progesterone-based protocols. Heifers in CIDR_PGF8 group (n = 10) received 2.0 mg i.m. estradiol benzoate (EB) and a new controlled internal drug release containing 1.9 g of progesterone (CIDR) on day 0 of study. At the time of CIDR withdrawal (day 8), heifers received an i.m. injection of PGF2α and 24 h later a second EB i.m. injection (0.5 mg). The three other groups received EB injections and CIDR insertion/withdrawal as aforementioned, except that an i.m. injection of PGF2α was administered on day 5. In addition, heifers in the CIDR_PGF5 group (n = 10) received a new CIDR, while heifers in theCIDR1x_PGF5 (n = 9) and CIDR2x_PGF5 (n = 11) groups received a previously used CIDR for 8 and 14 days, respectively. Heifers that received a PGF2α injection on day 5 showed lower circulating P4 than heifers treated on day 8 (CIDR_PGF5 = 1.98 ± 0.21 ng/ml; CIDR1x_PGF5 = 1.69 ± 0.17 ng/ml and CIDR2x_PGF5 = 1.33 ± 0.08 ng/ml versus CIDR_PGF8 = 3.31 ± 0.45 ng/ml). The dominant follicle (DF) growth rate was slower in those heifers receiving PGF2α injection on day 8 (CIDR_PGF8 = 0.72 ± 0.13 mm/day) than groups treated on day 5 (CIDR_PGF5 = 0.96 ± 0.12 mm/day; CIDR1x_PGF5 = 1.06 ± 0.15 mm/day and CIDR2x_PGF5 = 1.01 ± 0.06mm/day). In consequence, preovulatory follicle diameter on day 10 was smaller in those animals injected on day 8 (CIDR_PGF8 = 8.81 ± 6.7 mm) than in those treated on day 5 (CIDR_PGF5 = 10.00 ± 0.58 mm CIDR1x_PGF5 = 10.5 ± 0.69 mm and CIDR2x_PGF5 = 10.5 ± 0.35 mm). For heifers receiving PGF2α injection on day 5, no significant differences on plasma P4 concentrations, folliculargrowth rate and DF diameters were observed among heifers that received new or previously used CIDR inserts. These results suggest that the presence of corpus luteum during synchronization protocols is the main factor responsible for the increase in the plasma P4 concentrations and inhibition of DF growth.(AU)

Follicular growth and plasma progesterone patterns in Bos indicus x Bos taurus heifers submitted to different PGF2/progesterone-based synchronization protocols

The effect of different levels of progesterone (P4) concentrations on follicle growth and ovulatory capacity was evaluated in 40 crossbred Bos indicus x Bos taurus cyclic heifers submitted to distinct PGF2α + progesterone-based protocols. Heifers in CIDR_PGF8 group (n = 10) received 2.0 mg i.m. estradiol benzoate (EB) and a new controlled internal drug release containing 1.9 g of progesterone (CIDR) on day 0 of study. At the time of CIDR withdrawal (day 8), heifers received an i.m. injection of PGF2α and 24 h later a second EB i.m. injection (0.5 mg). The three other groups received EB injections and CIDR insertion/withdrawal as aforementioned, except that an i.m. injection of PGF2α was administered on day 5. In addition, heifers in the CIDR_PGF5 group (n = 10) received a new CIDR, while heifers in theCIDR1x_PGF5 (n = 9) and CIDR2x_PGF5 (n = 11) groups received a previously used CIDR for 8 and 14 days, respectively. Heifers that received a PGF2α injection on day 5 showed lower circulating P4 than heifers treated on day 8 (CIDR_PGF5 = 1.98 ± 0.21 ng/ml; CIDR1x_PGF5 = 1.69 ± 0.17 ng/ml and CIDR2x_PGF5 = 1.33 ± 0.08 ng/ml versus CIDR_PGF8 = 3.31 ± 0.45 ng/ml). The dominant follicle (DF) growth rate was slower in those heifers receiving PGF2α injection on day 8 (CIDR_PGF8 = 0.72 ± 0.13 mm/day) than groups treated on day 5 (CIDR_PGF5 = 0.96 ± 0.12 mm/day; CIDR1x_PGF5 = 1.06 ± 0.15 mm/day and CIDR2x_PGF5 = 1.01 ± 0.06mm/day). In consequence, preovulatory follicle diameter on day 10 was smaller in those animals injected on day 8 (CIDR_PGF8 = 8.81 ± 6.7 mm) than in those treated on day 5 (CIDR_PGF5 = 10.00 ± 0.58 mm CIDR1x_PGF5 = 10.5 ± 0.69 mm and CIDR2x_PGF5 = 10.5 ± 0.35 mm). For heifers receiving PGF2α injection on day 5, no significant differences on plasma P4 concentrations, folliculargrowth rate and DF diameters were observed among heifers that received new or previously used CIDR inserts. These results suggest that the presence of corpus luteum during synchronization protocols is the main factor responsible for the increase in the plasma P4 concentrations and inhibition of DF growth.