ABSTRACT The objective of this study was to evaluate the effects of cocoa meal in feed concentrate on the spermatogenesis in sheep. Twenty-five uncastrated males were divided into four groups and fed concentrated feed, supplemented with 0, 10, 20, and 30% cocoa meal, respectively, for 150 days. At the end of this period, the animals were slaughtered, the testicles were collected for histological slides, and testicular morphometric assessments were conducted, including cell type quantification in the seminiferous epithelium, intrinsic spermatogenesis yield, Sertoli-cell index, total number of Sertoli cells (TNSC), TNSC per testicular gram, daily sperm production (DSP), DSP per testicular gram, testicular sperm reserve (TSR), and TSR per testicular gram. Data were subjected to normality analysis using the Shapiro-Wilk test, followed by the tests for each condition, at a significance level of 5%. The administration of cocoa meal did not alter the germinal epithelial cell population, except for the number of type-A spermatogonia, which was lower in the control group than in the group that received 20% supplementation. A difference was found in the ratio between the number of type-A spermatogonia and primary spermatocytes in pre-leptotene/leptotene, and in the ratio between Sertoli cells and primary spermatocytes in pre-leptotene/leptotene, the control group and the group that received 10% cocoa meal significantly varied from each other, but not in relation to the other evaluated percentages. The TNSC, TNSC/g, DSP, and TSR levels were diminished in the group that consumed 30% cocoa meal. The TNSC, DSP, and TSR exhibited a negative quadratic tendency. Supplementation with 10% cocoa meal improved the TNSC, DSP, and TSR of sheep.

Abstract 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.

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.

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.

RESUMO 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.

ABSTRACT 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.

RESUMO 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 < 0,0001; 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.

ABSTRACT 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.

RESUMO O objetivo com este estudo foi verificar a acurácia da sexagem e da estimativa do número fetal em pequenos ruminantes por meio de exame ultrassonográfico. Foram avaliados 50 fetos de 36 ovelhas e 23 fetos de 11 cabras. No caso das ovelhas, 24, 10 e duas gestações foram simples, duplas e triplas, respectivamente. Enquanto nas cabras, três, cinco, duas e uma gestação foram simples, duplas, triplas e quádrupla, respectivamente. As avaliações foram realizadas aos 55 e 65 dias de gestação, por meio de exame ultrassonográfico transretal, utilizando-se um transdutor linear de dupla frequência (6,0 e 8,0 MHz). O sexo fetal foi diagnosticado pela localização do tubérculo genital ou visibilização da genitália externa. A acurácia da sexagem fetal foi avaliada por meio do teste Qui-quadrado (X2) considerando 5% de significância. O período gestacional interferiu (P<0,05) na acurácia da sexagem fetal. Obteve-se 30% e 82,61% de acurácia aos 55 dias de gestação, e 90% e 95,83% aos 65 dias, para cabras e ovelhas, respectivamente. O sexo fetal e o tipo de gestação não interferiram (P>0,05) na acurácia da sexagem em ambas as espécies nos períodos avaliados. Desta forma, a ultrassonografia em modo-B é um método eficiente para realização de diagnóstico precoce do sexo fetal e para determinação do número de fetos nos pequenos ruminantes no 65° dia da gestação, além disso, a acurácia da sexagem fetal não é influenciada pelo tipo de gestação, nem pelo sexo fetal.

SUMMARY The objective of this study was to verify the accuracy of sexing and fetal number estimation in small ruminants by ultrasonographic examination. Fifty fetuses from 36 sheep and 23 fetuses from 11 goats were evaluated. In the case of sheep, twenty-four, ten, and two pregnancy were single, double, and triple, respectively. Regarding the goats, three, five, two, and one simple pregnancy were single, double, triple, and quadruple, respectively. The evaluations were performed on days 55 and 65 of gestation, by means of transrectal ultrasonographic examination, using a dual-frequency linear transducer, (6.0 and 8.0 MHz). Fetal sex was diagnosed by the location of the genital tubercle or visualization of external genitalia. The accuracy of fetal sexing was evaluated using the Chi-square test (X2) considering a 5% significance level. The gestational period interfered in the accuracy of fetal sexing in both species (P <0.05). Obtained results showed 30% and 82.61% accuracy on day 55 of gestation, and 90% and 95.83% on day 65 for goats and sheep, respectively. The fetal sex and the type of pregnancy did not interfere in the sexing accuracy in both species during the evaluated periods (P> 0.05). Thus, B-mode ultrasonography is an efficient method to perform an early diagnosis of fetal sex and to determine the number of fetuses in the studied small ruminants on day 65 of gestation. In addition, the accuracy of fetal sexing is not influenced by the type of pregnancy or fetal sex.