Farmacopuntura em protocolos de sincronização de estro em pequenos ruminantes / Pharmacopuncture in estrus synchronization protocols in small ruminants

O objetivo com essa revisão foi realizar o levantamento da produção científica disponível e articular saberes a respeito do uso da farmacopuntura associada à protocolos de sincronização de estro principalmente em caprinos e ovinos, como também, em outras espécies de interesse. Ao analisar as pesquisas realizadas nos últimos 10 anos com a utilização de subdoses hormonais associada à farmacopuntura em programas de inseminação artificial em tempo fixo (IATF), nota-se que a farmacopuntura vem se tornando uma ferramenta promissora para viabilizar a redução das doses hormonais administradas e potencializar o efeito do fármaco aplicado, o que eleva a eficiência dos programas de sincronização de estro/ovulação ao reduzir os custos desses protocolos. Além de diminuir o custo desses protocolos para os produtores, nosso grupo de pesquisa realizou uma série de estudos nessa área, visando a busca por protocolos com menores doses hormonais, o que consequentemente reduz a quantidade de resíduos produzidos e promove manejos mais sustentáveis. Baseado nas publicações, pode-se afirmar que a farmacopuntura apresenta resultados satisfatórios e coloca-se como uma via de aplicação de sub-doses ou micro-doses hormonais em protocolos de controle do ciclo estral em caprinos e ovinos, como também, para outras espécies domésticas.(AU)

The objective of this review was to compile information regarding the use of pharmacopuncture associated with estrus synchronization protocols, mainly in goats and sheep, as well as in other species of interest, with the aim of updating and providing information on the subject. When analyzing the research carried out in the last 10 years with the use of hormonal underdoses associated with pharmacopuncture in fixed-time AI programs, these have become a promising tool to enable the reduction of administered hormonal doses and to potentiate the effect of the applied drug, the that increases the efficiency of estrus/ovulation synchronization programs by reducing the costs of these protocols. In addition to reducing the cost of these protocols for producers, our research group carried out a series of studies in this area, aiming at the search for protocols with lower hormonal doses, less waste produced and more sustainable management. Based on the publications, it can be stated that pharmacopuncture presents satisfactory results and is a way of applying sub-doses or micro-doses of hormones in protocols for controlling the estrous cycle in goats and sheep, as well as for other domestic species.(AU)

Renovação de diluidor na tecnologia de sêmen de pequenos ruminantes / Extender renewal in the technology of small ruminants semen

O objetivo desta revisão foi compilar o que se tem na literatura a respeito do efeito da renovação de diluidor seminal, mediante centrifugação, na qualidade do sêmen refrigerado de caprinos e ovinos e no tempo de viabilidade seminal. Um dos primeiros estudos publicados com essa metodologia foi realizado com sêmen de cão, em 2005, por Verstegen et al., seguido por estudos em outras espécies, como a equina e suína. Nosso grupo de pesquisa desenvolveu alguns estudos com diferentes metodologias para avaliar a eficiência do método, a necessidade do uso da centrífuga refrigerada nesse processo, o uso de antioxidantes no diluidor para renovação e o tempo de renovação do diluidor em pequenos ruminantes.(AU)

The objective of this revision was to compile what exists in literature regarding the effect of seminal diluent renewal, through centrifugation, in the quality of cooled semen of goat and sheep and during seminal viability time. One of the first studies published with this methodology was performed with dog semen in 2005 by Verstegen et al., followed by studies in other species, such as equine and swine. Our research group developed some studies using different methodologies to evaluate method efficiency, the need to use a cooled centrifuge in this process, the use of antioxidants in the diluent for renewal and the diluent renewal time in small ruminants.(AU)

Effect of different concentrations of Trolox® in association with docosahexaenoic acid on equine semen freezing

The aim of this study was to evaluate the association of different concentrations of Trolox® and the addition of a fixed concentration of DHA in the freezing of semen of Mangalarga Marchador stallions. To that end, 16 ejaculates were frozen in the following extenders: E1) BotuCrio® (BC; Control); E2) BC + 50 ngml-1 DHA + 30 µM Trolox® (BCDHA30T); E3) BC + 50 ngml-1 DHA + 40 µM Trolox® (BCDHA40T); E4) BC + 50 ngml-1 DHA + 50 µM Trolox® (BCDHA50T). All the tested extenders were similar in preserving different kinematic parameters, cell functional integrity, compacted DNA, and high and intermediate mitochondrial activity (P>0.05). However, sperm cryopreserved in BCDHA40T showed higher velocities than sperm frozen in the control extender (P<0.05). The 30 µM concentration of Trolox® was worse for sperm motility and the 50 µM concentration of Trolox® did not adequately preserve the structural integrity of the membranes in an extender containing DHA when compared to the BotuCrio® (P<0.05) extender. The use of Trolox® in freezing extenders containing DHA did not maximize the effect of BotuCrio®, except for in the case of sperm velocity parameters when at a concentration of 40 µM.(AU)

Testicular morphometry as a tool to evaluate the efficiency of immunocastration in lambs

This study aimed to evaluate the efficiency of immunocastration in lambs using testicular morphometry. Thirty lambs were randomly divided into two treatments (subcutaneous administration of 1.0 mL and 0.5 mL of an anti-GnRH vaccine) and a control group (1.0 mL saline solution). The animals were vaccinated at four months of age, received a second dose 30 days later, and were slaughtered 90 days after the first vaccine dose. After slaughter, testicles were collected, and samples were removed for histological processing and evaluation of testicular morphometric parameters. Analysis of variance, Tukey's test, and Kruskal-Wallis test were performed, with a 5% level of significance. There was a reduction in testicular weight, gonadosomatic index, seminiferous tubule diameter, germinal epithelium height, leydigosomatic index, and total tubule length. The total length per testicular gram increased in the immunocastrated group. Intrinsic spermatogenesis yield, Sertoli cell indices, and estimates of sperm and Sertoli cell production were reduced in the immunized groups (P < 0.05). The anti-GnRH vaccine in lambs at doses of 1.0 mL and 0.5 mL is sufficient to promote immunocastration, verified through severe changes in testicular morphometry from animals.(AU)

Docosahexaenoic acid in diluent for goat semen cryopreservation

Abstract The effects of docosahexaenoic acid (DHA) in the diluent for cryopreservation of goat semen on seminal quality and the optimal levels to be used were evaluated. After collection, semen was pooled and physically evaluated, then divided into four aliquots with different DHA levels in the diluent: 0, 10, 20, and 30 ng mL-1. The semen was cryopreserved in a TK 3000® freezing machine and then thawed for assessment at 37 °C. Sperm motility and vigor, membrane integrity, acrosomal integrity, mitochondrial activity, and sperm chromatin compaction were evaluated after thawing. A completely randomized design was used. For normally distributed variables, ANOVA and regression analysis were used to test for differences between treatments, and for non-parametric data, the Kruskal Wallis test was used at the 5% significance level. There were no differences among groups in terms of membrane integrity, acrosomal integrity, or chromatin compaction. There was a decrease in class I mitochondrial activity with increasing DHA level (P<0.05), but no differences in classes II, III, and IV (P>0.05). The inclusion of 10 to 30 ng mL-1 of DHA in the diluent did not result in improvements in seminal quality parameters after thawing, with some impairment observed in the mitochondrial activity of the sperm cells.

Application of hormonal subdoses at the Bai Hui acupoint for estrus synchronization in sheep / Aplicação de subdoses hormonais no ponto de acupuntura Bai Hui para sincronização de estro em ovelhas

Here, the efficiency of applying subdoses of equine chorionic gonadotropin (eCG) and prostaglandin F2α (PGF2α) at the Bai Hui (BH) acupoint for estrus synchronization in sheep was evaluated. Thirty Santa Inês ewes received intravaginal sponges containing 60 mg medroxyprogesterone acetate for 7 days. The animals were divided into five treatments (T) (n=6): T1 (control), 132.5 µg PGF2α (100% of dose) and 300 IU eCG (100% of dose), both intramuscularly (IM); T2, 39.75 µg PGF2α (30% of dose) at the BH acupoint and 300 IU eCG (100% of dose) IM; T3, 132.5 µg PGF2α (100% of dose) IM and 90 IU eCG (30% of dose) at the BH acupoint; T4, 39.75 µg PGF2α (30% of dose) and 90 IU eCG (30% of dose), both at the BH acupoint; and T5, 39.75 µg PGF2α (30% of dose) and 90 IU eCG (30% of dose) both at a false acupoint (IM, at the same location as in T1). After 12 h of sponge removal and hormone application, the ewes were subjected to monitoring for estrus and mating and ultrasound assessments of follicular growth and ovulation time every 12 h. The data were evaluated for normality and subjected to analysis of variance, considering a 5% probability. There were no differences in the percentage of animals in estrus (93.3±9.1%), interval from sponge removal to estrus onset (48.1±6.0 h), interval from sponge removal to estrus end (69.6±5.4 h), duration of estrus (24.0±4.7 h), interval from sponge removal to ovulation (73.7±5.9 h), interval from estrus onset to ovulation (26.5±2.3 h), size of the largest follicle (6.7±0.4 mm), follicular growth rate (0.8±0.1 mm•day-1), number of ovulations (1.3±0.1), and plasma progesterone levels 7 days after ovulation (9.6±2.0 ng•mL-1). The cost of the synchronization protocol per animal was US$ 4.37, 4.12, 2.96, 2.71, and 2.71 for T1, T2, T3, T4, and T5, respectively, with 37.99% cost reduction using PGF2α and eCG at 30% of the conventional doses. Therefore, the application of reduced PGF2α (39.75 µg) and eCG (90 IU) doses at the BH or false acupoint effectively synchronized estrus and reduced protocol cost in sheep, indicating that the conventional doses are overestimated.(AU)

Aqui, foi avaliada a eficiência da aplicação de subdoses de gonadotrofina coriônica equina (eCG) e prostaglandina F2α (PGF2α) no ponto de acupuntura Bai Hui (BH) para sincronização de estro em ovelhas. Trinta ovelhas Santa Inês receberam esponjas intravaginais contendo 60 mg de acetato de medroxiprogesterona por 7 dias. Os animais foram divididos em cinco tratamentos (T) (n=6): T1 (controle), 132,5 µg PGF2α (100% da dose) e 300 UI de eCG (100% da dose), ambos por via intramuscular (IM); T2, 39,75 µg de PGF2α (30% da dose) no ponto de acupuntura BH e 300 UI eCG (100% da dose) IM; T3, 132,5 µg PGF2α (100% da dose) IM e 90 UI eCG (30% da dose) no ponto de acupuntura BH; T4, 39,75 µg PGF2α (30% da dose) e 90 UI eCG (30% da dose), ambos no ponto de acupuntura BH; e T5, 39,75 µg PGF2α (30% da dose) e 90 UI eCG (30% da dose) ambos em um falso acuponto (IM, no mesmo local que em T1). Após 12 horas da retirada da esponja e aplicação do hormônio, as ovelhas foram submetidas ao monitoramento do estro e acasalamento, e avaliação ultrassonográfica do crescimento folicular e tempo de ovulação a cada 12 horas. Os dados foram avaliados quanto à normalidade e submetidos à análise de variância, considerando 5% de probabilidade. Não houve diferenças na porcentagem de animais em estro (93,3 ± 9,1%), intervalo da remoção da esponja ao início do estro (48,1 ± 6,0 h), intervalo da remoção da esponja ao final do estro (69,6 ± 5,4 h), duração do estro (24,0 ± 4,7 h), intervalo da remoção da esponja à ovulação (73,7 ± 5,9 h), intervalo do início do estro à ovulação (26,5 ± 2,3 h), tamanho do maior folículo (6,7 ± 0,4 mm), taxa de crescimento folicular (0,8 ± 0,1 mm•dia-1), número de ovulações (1,3 ± 0,1) e níveis de progesterona plasmática 7 dias após a ovulação (9,6 ± 2,0 ng•mL-1). O custo do protocolo de sincronização por animal foi de US $ 4,37, 4,12, 2,96, 2,71 e 2,71 para T1, T2, T3, T4 e T5, respectivamente, com redução de custo de 37,99% usando PGF2α e eCG a 30% das doses convencionais. Portanto, a aplicação de PGF2α (39,75 µg) e doses reduzidas de eCG (90 UI) no BH ou falso acuponto sincronizaram efetivamente o estro e reduziram o custo do protocolo em ovelhas, indicando que as doses convencionais estão superestimadas.(AU)

Docosahexaenoic acid in diluent for goat semen cryopreservation

The effects of docosahexaenoic acid (DHA) in the diluent for cryopreservation of goat semen on seminal quality and the optimal levels to be used were evaluated. After collection, semen was pooled and physically evaluated, then divided into four aliquots with different DHA levels in the diluent: 0, 10, 20, and 30 ng mL-1. The semen was cryopreserved in a TK 3000® freezing machine and then thawed for assessment at 37 °C. Sperm motility and vigor, membrane integrity, acrosomal integrity, mitochondrial activity, and sperm chromatin compaction were evaluated after thawing. A completely randomized design was used. For normally distributed variables, ANOVA and regression analysis were used to test for differences between treatments, and for non-parametric data, the Kruskal Wallis test was used at the 5% significance level. There were no differences among groups in terms of membrane integrity, acrosomal integrity, or chromatin compaction. There was a decrease in class I mitochondrial activity with increasing DHA level (P<0.05), but no differences in classes II, III, and IV (P>0.05). The inclusion of 10 to 30 ng mL-1 of DHA in the diluent did not result in improvements in seminal quality parameters after thawing, with some impairment observed in the mitochondrial activity of the sperm cells.(AU)

Docosahexaenoic acid associated vitamin E in the diluent for cryopreservation of goat semen / Ácido docosahexaenoico associado à vitamina E no meio diluidor para criopreservação de sêmen caprino

The objective of this study was to evaluate the effect of and determine the optimum level of inclusion of docosahexaenoic acid (DHA) in the diluent for goat semen cryopreservation. Five Boer males underwent semen collection, totaling 10 viable collections per animal. After evaluation, the ejaculates were pooled and fractionated in Tris-yolk medium with the addition of 0; 30; 45; or 60ng mL-1 of DHA and 0.4 mmol of alpha-tocopherol (Vitamin E). The semen was cryopreserved in a freezing machine (TK 3000TM) and placed in a cryogenic cylinder for subsequent analysis. Data were evaluated by regression analysis at 5% significance. There were no differences (P > 0.05) in sperm kinetic parameters evaluated by computer assisted sperm analysis: total motility (79.17 ± 17.31%), progressive motility (14.04 ± 5.73%), curvilinear speed (58.82 ± 6.35µm/s), progressive linear speed (22.49 ± 3.63µm/s), mean path speed (35.17 ± 4.52µm/s), linearity (38.69 ± 5.79%), rectilinearity (63.99 ± 6.64%), and oscillation index (59.68 ± 2.99%). There were no differences (P > 0.05) found from the membrane functional integrity test for reactive spermatozoa (69.66 ± 9.76%), plasma and acrosomal membrane integrity of intact spermatozoa (29.86 ± 7.57%), mitochondrial potential of Class I cryopreserved goat semen (72.75 ± 9.81%), and chromatin compaction of intact chromatin (96.87 ± 4.37%).(AU)

O estudo teve como objetivo avaliar o efeito e determinar o melhor nível de inclusão de ácido docosahexaenoico (DHA) no diluidor para criopreservação de sêmen caprino. Cinco machos Boer foram submetidos a coletas de sêmen, totalizando 10 coletas viáveis por animal. Após avaliação, os ejaculados foram agrupados (pool) e fracionados em meio Tris gema, acrescido de 0; 30; 45 e 60ng mL-1 de DHA e 0,4mmol de alfa-tocoferol. O sêmen foi criopreservado em máquina de congelamento TK 3000® e acondicionado em botijão criogênico para posterior análise. Os dados foram avaliados por análise de regressão a 5% de significância. Não houve diferença (P > 0,05) nos parâmetros cinéticos espermáticos avaliados pela análise assistida por computador: motilidade total (79,17 ± 17,31%), motilidade progressiva (14,04 ± 5,73%), velocidade curvilínea (58,82 ± 6,35µm/s), velocidade linear progressiva (22,49 ± 3,63µm/s), velocidade média do caminho (35,17 ± 4,52µm/s), linearidade (38,69 ± 5,79%), retilinearidade (63,99 ± 6,64%) e índice de oscilação (59,68 ± 2,99%). Não foram encontradas diferenças (P> 0,05) no teste de integridade funcional da membrana para espermatozoides reativos (69,66 ± 9,76%), integridade plasmática e membrana acrossomal dos espermatozoides intactos (29,86 ± 7,57%), potencial mitocondrial do sêmen caprino de classe I (72,75 ± 9,81%) e compactação da cromatina intacta (96,87 ± 4,37%). A inclusão de até 60ng mL-1 de DHA não promoveu melhora nos parâmetros de qualidade seminal de caprinos pós-descongelamento.(AU)

Docosahexaenoic acid associated vitamin E in the diluent for cryopreservation of goat semen / Ácido docosahexaenoico associado à vitamina E no meio diluidor para criopreservação de sêmen caprino

The objective of this study was to evaluate the effect of and determine the optimum level of inclusion of docosahexaenoic acid (DHA) in the diluent for goat semen cryopreservation. Five Boer males underwent semen collection, totaling 10 viable collections per animal. After evaluation, the ejaculates were pooled and fractionated in Tris-yolk medium with the addition of 0; 30; 45; or 60ng mL-1 of DHA and 0.4 mmol of alpha-tocopherol (Vitamin E). The semen was cryopreserved in a freezing machine (TK 3000TM) and placed in a cryogenic cylinder for subsequent analysis. Data were evaluated by regression analysis at 5% significance. There were no differences (P > 0.05) in sperm kinetic parameters evaluated by computer assisted sperm analysis: total motility (79.17 ± 17.31%), progressive motility (14.04 ± 5.73%), curvilinear speed (58.82 ± 6.35µm/s), progressive linear speed (22.49 ± 3.63µm/s), mean path speed (35.17 ± 4.52µm/s), linearity (38.69 ± 5.79%), rectilinearity (63.99 ± 6.64%), and oscillation index (59.68 ± 2.99%). There were no differences (P > 0.05) found from the membrane functional integrity test for reactive spermatozoa (69.66 ± 9.76%), plasma and acrosomal membrane integrity of intact spermatozoa (29.86 ± 7.57%), mitochondrial potential of Class I cryopreserved goat semen (72.75 ± 9.81%), and chromatin compaction of intact chromatin (96.87 ± 4.37%).

O estudo teve como objetivo avaliar o efeito e determinar o melhor nível de inclusão de ácido docosahexaenoico (DHA) no diluidor para criopreservação de sêmen caprino. Cinco machos Boer foram submetidos a coletas de sêmen, totalizando 10 coletas viáveis por animal. Após avaliação, os ejaculados foram agrupados (pool) e fracionados em meio Tris gema, acrescido de 0; 30; 45 e 60ng mL-1 de DHA e 0,4mmol de alfa-tocoferol. O sêmen foi criopreservado em máquina de congelamento TK 3000® e acondicionado em botijão criogênico para posterior análise. Os dados foram avaliados por análise de regressão a 5% de significância. Não houve diferença (P > 0,05) nos parâmetros cinéticos espermáticos avaliados pela análise assistida por computador: motilidade total (79,17 ± 17,31%), motilidade progressiva (14,04 ± 5,73%), velocidade curvilínea (58,82 ± 6,35µm/s), velocidade linear progressiva (22,49 ± 3,63µm/s), velocidade média do caminho (35,17 ± 4,52µm/s), linearidade (38,69 ± 5,79%), retilinearidade (63,99 ± 6,64%) e índice de oscilação (59,68 ± 2,99%). Não foram encontradas diferenças (P> 0,05) no teste de integridade funcional da membrana para espermatozoides reativos (69,66 ± 9,76%), integridade plasmática e membrana acrossomal dos espermatozoides intactos (29,86 ± 7,57%), potencial mitocondrial do sêmen caprino de classe I (72,75 ± 9,81%) e compactação da cromatina intacta (96,87 ± 4,37%). A inclusão de até 60ng mL-1 de DHA não promoveu melhora nos parâmetros de qualidade seminal de caprinos pós-descongelamento.

Reproductive parameters of lambs immunocastrated with anti-GnRH vaccine

Abstract The objective of this study was to evaluate the testicular biometric, seminal, and plasma testosterone levels in lambs subjected to an anti-GnRH vaccine as a method of castration. Thirty entire, crossbred Santa Inês male lambs were randomly distributed into three treatment (T): T1 was the control group, with the administration of 1 mL of saline solution subcutaneously (SC); 1.0 and 0.5 mL of an anti-GnRH vaccine were administered SC in T2 and T3, respectively. Testicular biometric variables, physical and morphological variables of semen, and plasma testosterone concentrations were evaluated. At D60, there was a reduction in testicular length, width, thickness, and scrotal circumference of the immunocastrated animals regardless of the vaccine dose used (P < 0.05). A reduction in semen physical variables at both dosages (P < 0.05) was observed, with azoospermia, in 80% and 70% of animals in the T2 and T3 groups, respectively. At D60, the immunocastrated animals also showed an increase in spermatozoa defects (P < 0.05), whereas plasma testosterone concentration decreased (P < 0.05). Immunocastration of lambs using the Bopriva vaccine at doses of 1.0 and 0.5 mL is efficient in inducing azoospermia in up to 80% of animals, although two doses in a 30-day interval are necessary for it to be an effective and safe method. Efficacy was demonstrated through a reduction in serum testosterone levels, testicular biometry, and seminal fluid analysis. Considering the efficacy of both doses in this study, we recommend using the lower dose (0.5 mL), which will allow for a 50% reduction in vaccine costs.

Reproductive parameters of lambs immunocastrated with anti-GnRH vaccine

The objective of this study was to evaluate the testicular biometric, seminal, and plasma testosterone levels in lambs subjected to an anti-GnRH vaccine as a method of castration. Thirty entire, crossbred Santa Inês male lambs were randomly distributed into three treatment (T): T1 was the control group, with the administration of 1 mL of saline solution subcutaneously (SC); 1.0 and 0.5 mL of an anti-GnRH vaccine were administered SC in T2 and T3, respectively. Testicular biometric variables, physical and morphological variables of semen, and plasma testosterone concentrations were evaluated. At D60, there was a reduction in testicular length, width, thickness, and scrotal circumference of the immunocastrated animals regardless of the vaccine dose used (P < 0.05). A reduction in semen physical variables at both dosages (P < 0.05) was observed, with azoospermia, in 80% and 70% of animals in the T2 and T3 groups, respectively. At D60, the immunocastrated animals also showed an increase in spermatozoa defects (P < 0.05), whereas plasma testosterone concentration decreased (P < 0.05). Immunocastration of lambs using the Bopriva vaccine at doses of 1.0 and 0.5 mL is efficient in inducing azoospermia in up to 80% of animals, although two doses in a 30-day interval are necessary for it to be an effective and safe method. Efficacy was demonstrated through a reduction in serum testosterone levels, testicular biometry, and seminal fluid analysis. Considering the efficacy of both doses in this study, we recommend using the lower dose (0.5 mL), which will allow for a 50% reduction in vaccine costs.(AU)

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

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

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

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

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

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

Subdose of human chorionic gonadotropin applied at the Hou Hai acupoint on follicular dynamics and luteal development in donkeys

The objective of this study was to evaluate the effects of an hCG subdose applied at the Hou Hai acupoint as an ovulation inducer in donkeys. Eleven donkeys were distributed in randomized blocks in T1 = application of 1,500 IU of hCG intravenous (IV); T2 = 450 IU of hCG applied at the false acupoint (IV), and T3 = 450 IU of hCG applied at the Hou Hai acupoint. There was no difference (P > 0.05) between the treatments regarding the mean diameter of the pre-ovulatory follicle (34.5 ± 1.3 mm), the ovulation rate (96.97%), the interval between induction and ovulation (58.07 ± 16.82 h), the mean diameter of the CL (D0 = 23.0 ± 0.6; D2 = 27.7 ± 1.9 and D8 = 28.2 ± 0.8mm), and serum P4 concentrations (10.50 ± 2.99 ng.mL-1). The application of 450 IU of hCG at the Hou Hai acupoint increased ovulation rate (72.73%) more than 48 h after induction (P = 0.03) and a larger diameter of the CL on D4 (30.7 ± 5.1 mm) (P = 0.04). The vascularization area of the CL on D8, obtained by minimum number of colored pixel (NCP), was greater (P 0.05) in the donkeys that received 1,500 IU of IV hCG (T1, 41.91 ± 1.17), and we found a positive correlation (P 0.05) between mean NCP and P4 concentration in the donkeys that received 450 IU of hCG IV at the false acupoint (T2) or at the Hou Hai acupoint (T3). The application of 450 IU of hCG by IV route at the false acupoint or the Hou Hai acupoint was sufficient to induce ovulation in donkeys, demonstrating that the average dosage commonly used for this species is too high.

Subdose of human chorionic gonadotropin applied at the Hou Hai acupoint on follicular dynamics and luteal development in donkeys

The objective of this study was to evaluate the effects of an hCG subdose applied at the Hou Hai acupoint as an ovulation inducer in donkeys. Eleven donkeys were distributed in randomized blocks in T1 = application of 1,500 IU of hCG intravenous (IV); T2 = 450 IU of hCG applied at the false acupoint (IV), and T3 = 450 IU of hCG applied at the Hou Hai acupoint. There was no difference (P > 0.05) between the treatments regarding the mean diameter of the pre-ovulatory follicle (34.5 ± 1.3 mm), the ovulation rate (96.97%), the interval between induction and ovulation (58.07 ± 16.82 h), the mean diameter of the CL (D0 = 23.0 ± 0.6; D2 = 27.7 ± 1.9 and D8 = 28.2 ± 0.8mm), and serum P4 concentrations (10.50 ± 2.99 ng.mL-1). The application of 450 IU of hCG at the Hou Hai acupoint increased ovulation rate (72.73%) more than 48 h after induction (P = 0.03) and a larger diameter of the CL on D4 (30.7 ± 5.1 mm) (P = 0.04). The vascularization area of the CL on D8, obtained by minimum number of colored pixel (NCP), was greater (P 0.05) in the donkeys that received 1,500 IU of IV hCG (T1, 41.91 ± 1.17), and we found a positive correlation (P 0.05) between mean NCP and P4 concentration in the donkeys that received 450 IU of hCG IV at the false acupoint (T2) or at the Hou Hai acupoint (T3). The application of 450 IU of hCG by IV route at the false acupoint or the Hou Hai acupoint was sufficient to induce ovulation in donkeys, demonstrating that the average dosage commonly used for this species is too high.(AU)

Spermatogenesis in sheep supplemented with detoxified castor bean (Ricinus communis L. ) as a replacement for soybean meal / Espermatogênese de ovinos suplementados com torta de mamona (Ricinus communis L. ) destoxificada em substituição ao farelo de soja

The objective was to evaluate the effect of replacing soybean meal with the detoxified castor bean cake on testicular morphometry and spermatogenesis of sheep. Were used 24 uncastrated, 9-month old sheep weighing 29±0.8 kg they were randomly distributed among three treatments: T1 = 0%, T2 = 50%, and T3 = 100% substitution of soybean meal with detoxified castor bean cake. The animals were fed with Aruana grass pastage (Panicum maximum Aruana) and a ration for 90 days. After slaughtering, the testicles were collected and histological slides were prepared with tissue fragments. The data were evaluated for normality using the Shapiro-Wilk test, and analysis of variance was carried out at 5% level of significance. Substitution of soybean meal with detoxified castor bean cake had no effect on any of the assessed variables at the tested levels (P >0.05). The mean yield of spermatogenesis was 72.91 rounded spermatids per spermatogonium; the mean of total number of germ cells held by a Sertoli cell was 12.09; the mean of the testicular spermatic reserve was 31.82×109 and that per testicular gram was 238.28×106; the mean of daily spermatic production was 3.03×109 and that per testicular gram was 22.69×106; and the total number of Sertoli cells was 4.15×109 and that per testicular gram was 34.51×106. The results show that it is possible to replace 100% of the soybean meal with detoxified castor bean cake in sheep diet without any effects on spermatogenesis; however, it is important to perform seminal evaluations in future studies.(AU)

O objetivo com este estudo foi avaliar o efeito da substituição do farelo de soja pela torta de mamona destoxificada sobre a espermatogênese em ovinos. Foram utilizados 24 ovinos, com peso médio de 29±0,8kg e 9 meses de idade, distribuídos aleatoriamente em três tratamentos: T1= 0%, T2= 50% e T3= 100% de substituição do farelo de soja pela torta de mamona destoxificada. Os animais foram alimentados com pastagem de capim Aruana (Panicum maximum cv. Aruana) e ração durante 90 dias. Por ocasião do abate, coletou-se os testículos para mensurações e avaliação da espermatogênese. Os dados foram avaliados quanto à normalidade por meio do teste de Shapiro-Wilk e utilizou-se a Análise de Variância com 5% de significância. Não houve efeito da substituição do farelo de soja pela torta de mamona destoxificada nos níveis testados (P>0,05) para nenhuma das variáveis avaliadas. O rendimento médio da espermatogênese foi de 72,91 espermátides arredondadas produzidas a partir de uma espermatogônia; a média do número total de células germinativas sustentadas por uma de célula de Sertoli foi 12,09; a média da reserva espermática testicular total foi de 31,82x109 e de 238,28x106 por grama de testículo; a média da produção espermática diária foi de 3,03x109 e de 22,69x106 por grama de testículo e o número total de células de Sertoli foi de 4,15x109 e de 34,51x106 por grama de testículo. Os resultados demonstram que é possível substituir 100% do farelo de soja pela torta de mamona destoxificada na dieta de ovinos sem prejuízos no processo de espermatogênese.(AU)

Spermatogenesis in sheep supplemented with detoxified castor bean (Ricinus communis L. ) as a replacement for soybean meal / Espermatogênese de ovinos suplementados com torta de mamona (Ricinus communis L. ) destoxificada em substituição ao farelo de soja

The objective was to evaluate the effect of replacing soybean meal with the detoxified castor bean cake on testicular morphometry and spermatogenesis of sheep. Were used 24 uncastrated, 9-month old sheep weighing 29±0.8 kg they were randomly distributed among three treatments: T1 = 0%, T2 = 50%, and T3 = 100% substitution of soybean meal with detoxified castor bean cake. The animals were fed with Aruana grass pastage (Panicum maximum Aruana) and a ration for 90 days. After slaughtering, the testicles were collected and histological slides were prepared with tissue fragments. The data were evaluated for normality using the Shapiro-Wilk test, and analysis of variance was carried out at 5% level of significance. Substitution of soybean meal with detoxified castor bean cake had no effect on any of the assessed variables at the tested levels (P >0.05). The mean yield of spermatogenesis was 72.91 rounded spermatids per spermatogonium; the mean of total number of germ cells held by a Sertoli cell was 12.09; the mean of the testicular spermatic reserve was 31.82×109 and that per testicular gram was 238.28×106; the mean of daily spermatic production was 3.03×109 and that per testicular gram was 22.69×106; and the total number of Sertoli cells was 4.15×109 and that per testicular gram was 34.51×106. The results show that it is possible to replace 100% of the soybean meal with detoxified castor bean cake in sheep diet without any effects on spermatogenesis; however, it is important to perform seminal evaluations in future studies.

O objetivo com este estudo foi avaliar o efeito da substituição do farelo de soja pela torta de mamona destoxificada sobre a espermatogênese em ovinos. Foram utilizados 24 ovinos, com peso médio de 29±0,8kg e 9 meses de idade, distribuídos aleatoriamente em três tratamentos: T1= 0%, T2= 50% e T3= 100% de substituição do farelo de soja pela torta de mamona destoxificada. Os animais foram alimentados com pastagem de capim Aruana (Panicum maximum cv. Aruana) e ração durante 90 dias. Por ocasião do abate, coletou-se os testículos para mensurações e avaliação da espermatogênese. Os dados foram avaliados quanto à normalidade por meio do teste de Shapiro-Wilk e utilizou-se a Análise de Variância com 5% de significância. Não houve efeito da substituição do farelo de soja pela torta de mamona destoxificada nos níveis testados (P>0,05) para nenhuma das variáveis avaliadas. O rendimento médio da espermatogênese foi de 72,91 espermátides arredondadas produzidas a partir de uma espermatogônia; a média do número total de células germinativas sustentadas por uma de célula de Sertoli foi 12,09; a média da reserva espermática testicular total foi de 31,82x109 e de 238,28x106 por grama de testículo; a média da produção espermática diária foi de 3,03x109 e de 22,69x106 por grama de testículo e o número total de células de Sertoli foi de 4,15x109 e de 34,51x106 por grama de testículo. Os resultados demonstram que é possível substituir 100% do farelo de soja pela torta de mamona destoxificada na dieta de ovinos sem prejuízos no processo de espermatogênese.

Does resveratrol preserve the integrity of the sperm membrane in goats after thawing? / O resveratrol preserva a integridade da membrana espermática de caprinos após descongelamento?

This study aimed to determine the effects of resveratrol in the trisaminomethane (TRIS)-egg yolk extender and its optimal inclusion level for goat semen cryopreservation. Five ejaculates of three Anglo Nubian goats were used, each divided into four 200 µL aliquots for use in four treatments: 0.00 (control), 0.04, 0.08 and 0.12 mg mL-¹ resveratrol in the TRIS-egg yolk extender. We evaluated progressive sperm motility and sperm vigor post-dilution, post-cooling, and post thawing; membrane integrity (HOST); and acrosomal integrity and performed a slow thermoresistance test (STT). The data were submitted to a regression analysis at a 5% probability. There was no difference in progressive motility or sperm vigor in the post-dilution (89.5, 89.0, 88.7 and 88.3, and 4.9, 5.0, 4.9, and 4.9) or post-cooling (81.0, 82.0, 83.0, and 78.3; and 4.3, 4.3, 4.2, and 4.2) experiments (P > 0.05), or in the complementary acrosomal integrity test (42.0, 47.4, 42.2 and 38.2) (P > 0.05). However, the motility and vigor parameters decreased linearly in the post-thaw phase, as well as during the 2 hours of incubation on STT (P < 0.05). These factors increased quadratically when resveratrol was added to HOST, to an optimal level of 0.039 mg mL-¹ resveratrol for a plasma membrane integrity of 52.55% (P < 0.05). The inclusion of resveratrol was effective in maintaining sperm viability; in particular, it was effective in maintaining plasmatic membrane integrity during the cryopreservation process up to 0.039 mg mL-1, meaning that it could be an alternative to conventionally used seminal extenders in goats.

O objetivo deste estudo foi determinar os efeitos e o melhor nível de inclusão de resveratrol em meio diluidor TRIS gema de ovo para criopreservação de sêmen caprino. Foram utilizados cinco ejaculados de três bodes da raça Anglo Nubiana. Cada ejaculado foi dividido em quatro alíquotas de 200 µL, compondo quatro tratamentos: 0,00 (controle); 0,04; 0,08 e 0,12mg mL-¹ de resveratrol no diluidor TRIS gema de ovo. Foram avaliadas a motilidade espermática progressiva e o vigor espermático pós diluição, pós-resfriamento e pós-descongelamento; integridade de membrana (HOST); integridade acrossomal e teste de termorresistência lento (TTR). Os dados foram submetidos à análise de regressão a 5% de probabilidade. Não houve diferença para a motilidade progressiva e o vigor espermático, respectivamente, na pós-diluição (89,5; 89,0; 88,7 e 88,3) e (4,9; 5,0; 4,9 e 4,9) e pós-resfriamento (81,0; 82,0; 83,0 e 78,3) e (4,3; 4,3; 4,2 e 4,2) (P 0,05). No entanto, os parâmetros motilidade e vigor reduziram linearmente no pós-descongelamento, assim como durante as 2 horas de incubação no TTR (P < 0,05). Houve efeito quadrático com a inclusão de resveratrol para HOST, apresentando um nível ótimo de 0,039mg mL-¹ de resveratrol para integridade de membrana plasmática de 52,55% (P <0,05). A inclusão de resveratrol foi eficiente na manutenção da viabilidade espermática; em especial, foi eficiente na manutenção da integridade da membrana plasmática durante o processo de criopreservação até o nível de 0,039mg mL-¹, podendo ser uma alternativa na composição dos diluidores seminais de caprinos.

Does resveratrol preserve the integrity of the sperm membrane in goats after thawing? / O resveratrol preserva a integridade da membrana espermática de caprinos após descongelamento?

This study aimed to determine the effects of resveratrol in the trisaminomethane (TRIS)-egg yolk extender and its optimal inclusion level for goat semen cryopreservation. Five ejaculates of three Anglo Nubian goats were used, each divided into four 200 µL aliquots for use in four treatments: 0.00 (control), 0.04, 0.08 and 0.12 mg mL-¹ resveratrol in the TRIS-egg yolk extender. We evaluated progressive sperm motility and sperm vigor post-dilution, post-cooling, and post thawing; membrane integrity (HOST); and acrosomal integrity and performed a slow thermoresistance test (STT). The data were submitted to a regression analysis at a 5% probability. There was no difference in progressive motility or sperm vigor in the post-dilution (89.5, 89.0, 88.7 and 88.3, and 4.9, 5.0, 4.9, and 4.9) or post-cooling (81.0, 82.0, 83.0, and 78.3; and 4.3, 4.3, 4.2, and 4.2) experiments (P > 0.05), or in the complementary acrosomal integrity test (42.0, 47.4, 42.2 and 38.2) (P > 0.05). However, the motility and vigor parameters decreased linearly in the post-thaw phase, as well as during the 2 hours of incubation on STT (P < 0.05). These factors increased quadratically when resveratrol was added to HOST, to an optimal level of 0.039 mg mL-¹ resveratrol for a plasma membrane integrity of 52.55% (P < 0.05). The inclusion of resveratrol was effective in maintaining sperm viability; in particular, it was effective in maintaining plasmatic membrane integrity during the cryopreservation process up to 0.039 mg mL-1, meaning that it could be an alternative to conventionally used seminal extenders in goats.(AU)

O objetivo deste estudo foi determinar os efeitos e o melhor nível de inclusão de resveratrol em meio diluidor TRIS gema de ovo para criopreservação de sêmen caprino. Foram utilizados cinco ejaculados de três bodes da raça Anglo Nubiana. Cada ejaculado foi dividido em quatro alíquotas de 200 µL, compondo quatro tratamentos: 0,00 (controle); 0,04; 0,08 e 0,12mg mL-¹ de resveratrol no diluidor TRIS gema de ovo. Foram avaliadas a motilidade espermática progressiva e o vigor espermático pós diluição, pós-resfriamento e pós-descongelamento; integridade de membrana (HOST); integridade acrossomal e teste de termorresistência lento (TTR). Os dados foram submetidos à análise de regressão a 5% de probabilidade. Não houve diferença para a motilidade progressiva e o vigor espermático, respectivamente, na pós-diluição (89,5; 89,0; 88,7 e 88,3) e (4,9; 5,0; 4,9 e 4,9) e pós-resfriamento (81,0; 82,0; 83,0 e 78,3) e (4,3; 4,3; 4,2 e 4,2) (P < 0,05), assim como para o teste complementar integridade acrossomal (42,0; 47,4; 42,2 e 38,2) (P > 0,05). No entanto, os parâmetros motilidade e vigor reduziram linearmente no pós-descongelamento, assim como durante as 2 horas de incubação no TTR (P < 0,05). Houve efeito quadrático com a inclusão de resveratrol para HOST, apresentando um nível ótimo de 0,039mg mL-¹ de resveratrol para integridade de membrana plasmática de 52,55% (P <0,05). A inclusão de resveratrol foi eficiente na manutenção da viabilidade espermática; em especial, foi eficiente na manutenção da integridade da membrana plasmática durante o processo de criopreservação até o nível de 0,039mg mL-¹, podendo ser uma alternativa na composição dos diluidores seminais de caprinos.(AU)

Application of low dose of equine chorionic gonadotropin at acupoint Hou Hai for fixed-time artificial insemination in beef cows / Aplicação de baixa dose de gonadotrofina coriônica equina no acuponto Hou Hai em protocolos de inseminação artificial em tempo fixo para vacas de corte

This study evaluated the efficiency application of low dose of equine chorionic gonadotropin (eCG) using Hou Hai acupoint for fixed-term artificial insemination (FTAI) in beef cows. Seventy cows received intravaginal devices with progesterone, and 3 mg of estradiol benzoate on day zero (D0) of FTAI. On D9, the devices were removed, and 150 μg of prostaglandin F2α was applied; the animals were then randomly distributed to three treatment groups: T1 (n = 23), 300 IU of eCG intramuscularly (IM); T2 (n = 23), 90 IU of eCG at acupoint Hou Hai; and T3 (n = 25), 90 IU of eCG at false acupoint IM. On D10, the animals received 1 mg of gonadotropin-releasing hormone IM, and FTAI was performed, 52 h after the devices were removed. The quantitative variables with normal distribution were assessed using analysis of variance at 5% probability. For variables that did not show normal distribution, Kruskal-Wallis test at 5% probability was used. No significant difference was noted among the groups (P > 0.05) for the interval between device withdrawal at ovulation (58.2 ± 12.2 h), diameter of the largest follicle on D9 (9.9 ± 2.2 mm), diameters of the ovulatory follicle (12.2 ± 3.0 mm) and the second largest follicle (6.7 ± 2.1 mm), follicular growth rate (0.8 ± 0.3 mm/d), ovulation rate (82%), corpus luteum size (2.32 ± 0.35 cm2), and pregnancy rate (58.67%). The protocol cost per animal was US$ 10.67 (T1) and US$ 8.50 (T2 and T3). The use of 90 IU of eCG applied at Hou Hai acupoint or at false acupoint caused satisfactory synchronization of estrus in beef cows. In addition, this procedure was cost-effective.(AU)

Avaliou-se a eficiência da aplicação de subdose gonadotrofina coriônica equina (eCG) utulizando como via de administração o acuponto Hou Hai em protocolos de inseminação artificial em tempo fixo (IATF) para vacas de corte. Setenta vacas receberam no dia zero (D0) do protocolo dispositivos intravaginais com progesterona, e 3mg de benzoato de estradiol. No D9 foram retirados os dispositivos, aplicado 150µg de prostaglandina F2α e distribuídos os animais aleatoriamente em três tratamentos: T1 (n=23) - 300UI de eCG por via intra muscular (IM); T2 (n=23) - 90UI de eCG no acuponto Hou Hai e T3 (n=25) - 90UI de eCG em falso acuponto, IM. No D10 os animais receberam 1mg de GnRH por via IM e foi realizada a IATF, 52 horas após a retirada dos dispositivos. Para as variáveis quantitativas com distribuição normal foi utilizada ANOVA, a 5% de probabilidade. Para as variáveis que não apresentaram distribuição normal, utilizou-se o teste de Kruskal-Wallis, a 5% de probabilidade. Não houve diferença (P >0,05) para o intervalo entre a retirada do dispositivo à ovulação (58,2 ± 12,2h), diâmetro do maior folículo no D9 (9,9 ± 2,2mm), diâmetros do folículo ovulatório (12,2 ± 3,0mm) e do segundo maior folículo (6,7 ± 2,1mm), taxa de crescimento folicular (0,8 ± 0,3mm/d), taxa de ovulação (82%), área de corpo lúteo (2,32 ± 0,35 cm2) e taxa de prenhez (58,67%). O custo do protocolo por animal foi de US$ 10,67 (T1) e US$ 8,50 (T2 e T3). Sugere-se que a utilização de 90UI de eCG aplicada no acuponto Hou Hai ou em falso acuponto foram satisfatórios na sincronização de estro de vacas de corte. Além disso, possibilita a redução do custo do protocolo.(AU)