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. sheep Twentyfive Twenty five 0 10 20 30 respectively 15 days period slaughtered slides conducted epithelium yield Sertolicell index , (TNSC) gram (DSP) TSR, (TSR) ShapiroWilk Shapiro Wilk test condition 5 5% population typeA supplementation preleptotene/leptotene, preleptoteneleptotene pre leptotene/leptotene, leptotene pre-leptotene/leptotene percentages TNSCg g TNSC/g tendency 1 2 3 (TNSC (DSP (TSR preleptotene preleptotene/leptotene leptoteneleptotene leptotene/leptotene

Resumen Diversas semillas de oleaginosas son fuente de ácidos grasos esenciales indispensables para cubrir las funciones básicas en el organismo. El estudio tuvo como objetivo evaluar el efecto de la inclusión de semilla de linaza (Linum usitatissimum) en la dieta de conejos por medio del perfil metabólico. Se utilizaron 20 machos de la raza Nueva Zelandia de 8 meses de edad. Distribuidos en cuatro grupos, recibieron niveles de semilla de linaza (L. usitatissimum) en la dieta total: 0 %, 3 %, 6 % y 9 %, durante un período de 88 días. Los animales se sometieron a colecta de sangre cada 22 días para determinar glucosa, colesterol total, lipoproteínas de alta densidad (HDL), triglicéridos, lipoproteínas de muy baja densidad (VLDL), lipoproteínas de baja densidad (LDL), creatinina y urea. Los datos se evaluaron por análisis de varianza y de regresión con 5 % de significancia. La inclusión de semilla de linaza (L. usitatissimum) en la dieta presentó comportamiento cuadrático positivo para concentración plasmática de glucosa y colesterol total. No hubo diferencia significativa entre los tratamientos para la concentración plasmática de triglicéridos, lipoproteínas de baja densidad (LDL), lipoproteínas de muy baja densidad (VLDL) urea. Hubo comportamiento cúbico para lipoproteínas de alta densidad (HDL) y creatinina. La inclusión de linaza (L. usitatissimum) en la dieta no presentó efecto hipercolesterolémico en estos animales.

Abstract Many oilseeds are a source of essentials fatty acids that are necessary for the coverage of the basic functions in the organism. The study aimed to evaluate the effect of flaxseed (Linum usitatissimum) inclusion in the diet of rabbits through the metabolic profile. Twenty males of the New Zealand breed of 8 months were used. Divided in four groups, they received flaxseed (L. usitatissimum) levels in the total diet: 0 %, 3 %, 6 % y 9 %, for a period of 88 days. The animals were subjected to a blood collection every 22 days to determine glucose, total cholesterol, high density lipoprotein (HDL), triglycerides, very low-density lipoprotein (VLDL), low density lipoprotein (LDL), creatinine and urea. The data were evaluated by analysis of variance and regression with 5 % significance. The inclusion of flaxseed (L. usitatissimum) in the rabbit diet showed a positive quadratic effect for plasmatic concentration of glucose and total cholesterol. There was no significant difference between treatments for plasma concentration of triglycerides, low- density lipoproteins (LDL), very low - density lipoproteins (VLDL) and urea. There was a cubic effect for cholesterol- and creatinine. The inclusion of flaxseed (L. usitatissimum) in the diet had hypercholesterolemic effect in these animals.

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 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.

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.

Resumen El objetivo de este trabajo fue evaluar el efecto de la semilla de linaza (Linum usitatissimum) en la calidad física y morfológica del semen fresco y refrigerado de conejos. Se emplearon 20 machos Nueva Zelandia distribuidos en grupos según niveles de inclusión de semilla en su dieta durante 100 días: 0%, 3%, 6% y 9%. 15 días después de iniciar el suministro de la dieta se realizaron colectas seminales una vez por semana con vagina artificial y una hembra como maniquí. Para la evaluación física y morfológica se diluyó en medio Tris-yema y refrigerado a 5 °C; se valoró la motilidad espermática progresiva, el vigor espermático y la morfología espermática a las 0, 2, 12, 24, 48 y 72 horas post-refrigeración. La información fue analizada mediante análisis de varianza y de regresión al 5% de significancia. Para la morfología, volumen, color y aspecto seminal del semen fresco (p > 0,05) no hubo diferencia. Para volumen, turbidez, motilidad, vigor y concentración, hubo valores medios de 0,92 ± 0,24 mL; 0,86 ± 0,35; 76,58 ± 7,13%; 3,37 ± 0,28 y 205,75 ± 20,77 × 106 sptz, respectivamente. Hubo diferencia para motilidad y vigor espermático a las 0, 2, 12, 24, 48 y 72 horas post-refrigeración (p < 0,005). Se concluyó que la inclusión de hasta 9% de semilla de linaza en la dieta de conejos no alteró los parámetros físicos y microscópicos evaluados en este estudio en semen fresco y refrigerado de conejo.

Abstract The aim of this study was to evaluate the effect of flaxseed (Linum usitatissimum) on the physical and morphological quality of fresh and refrigerated rabbit semen. Twenty New Zealand males were used, and distributed in four groups according to the level of inclusion of flaxseed in the total diet for 100 days: 0 %, 3 %, 6 %, and 9 %. Fifteen days after starting the supply of the diet, seminal collections were made once a week with an artificial vagina and a female as a dummy. For the physical and morphological evaluations, the semen was diluted in Tris-Egg Yolk medium and cooling at 5 °C; progressive sperm motility, and spermatic vigor and morphology were assessed at 0, 2, 12, 24, 48, and 72 post-cooling hours. This information was analyzed through analyses of variance and regression at 5 % significance. For the morphology, volume, color, and seminal appearance of fresh semen (p > 0.05), there was no difference. For volume, turbidity, motility, vigor, and concentration, average values of 0.92 ± 0.24 mL; 0.86 ± 0.35; 76.58 ± 7.13 %; 3.37 ± 0.28, and 205.75 ± 20.77 x 106 sptz were recorded, respectively. There were differences for motility and vigor at 0, 2, 12, 24, 48, and 72 post-cooling hours (p > 0.005). In conclusion, the inclusion of up to 9 % flaxseed in the rabbit diet did not alter the physical and microscopic parameters evaluated in this study in fresh and refrigerated rabbit semen.

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 com o estudo foi avaliar o efeito da inclusão de semente de linhaça na dieta de machos caprinos sobre a resistência do sêmen frente ao processo de criopreservação. Foram utilizados 16 machos, os quais foram distribuídos em quatro grupos e receberam dieta suplementada com 0, 4, 8 e 12% de semente de linhaça, durante um período de 60 dias. As coletas de sêmen iniciaram 25 dias após o início do fornecimento das dietas. Apenas os ejaculados que apresentaram motilidade e vigor acima de 70% e 3, respectivamente, foram encaminhados para o processo de criopreservação. Após o descongelamento, o sêmen foi avaliado através dos testes de termorresistência, hiposmótico e de integridade acrossomal. Os dados foram submetidos à análise de variância e regressão a 5% de significância. Houve comportamento quadrático positivo para a motilidade progressiva aos 60, 120 e 180 minutos no teste de termoresistência (TTR), e comportamento quadrático positivo para o vigor espermático pós-descongelamento aos 120 e 180 minutos no TTR (P<0,05). Entretanto, obteve-se comportamento quadrático negativo para a integridade da membrana plasmática pelo Teste Hiposmótico (P<0,05) e houve diferença para o teste de integridade acrossomal, apresentando nível máximo ótimo de 3,25% de semente de linhaça para integridade acrossomal de 65,83% (P<0,05). Os resultados obtidos demonstram que a adição de 12% de semente de linhaça na dieta de reprodutores caprinos melhora a qualidade espermática pós-descongelamento.

ABSTRACT The objective of this study was to evaluate the effects of the inclusion of flaxseed in the diet of male goats on the resistance of semen to the cryopreservation process. Sixteen males were distributed in four groups and fed a diet supplemented with 0, 4, 8, and 12% of flaxseed for a period of 60 days. Only the ejaculates that presented motility and vigor above 70% and 3, respectively, were sent through the cryopreservation process. After thawing, the semen was evaluated through thermo resistance, hypoosmotic, and acrossomal integrity tests. The data were submitted to analysis of variance and regression at 5% of significance. There was positive quadratic behavior for motility after 60, 120, and 180 min in the thermoresistance test (TTR), and positive quadratic behavior for sperm vigor after thawing after 120 and 180 min in the TTR (P < 0.05). However, negative quadratic behavior was obtained for plasma membrane integrity according to the hypoosmotic test (P < 0.05) and there was a difference in the acrosomal integrity test, presenting an optimum maximum level of 3.25% of flaxseed for acrosomal integrity of 65.83% (P < 0.05). The results obtained demonstrated that the addition of as much as 12% of flaxseed to the diet of goat breeders improved post-thawing sperm quality.

The substitution of equine chorionic gonadotropin (eCG) by follicle-stimulating hormone (FSH) in protocols for synchronization of ovulation in Santa Inês ewes was assessed. Ten females were submitted to the insertion of intravaginal sponges containing 60 mg medroxyprogesterone acetate for 10 days; after this period sponges were withdrawn and the animals were randomly divided into two groups. Group 1 (n = 5): intramuscular injection of 0.5 mL d-cloprostenol and 300 UI eCG; Group 2 (n = 5): intramuscular injection of 0.5 mL d-cloprostenol and 20 mg FSH. Interval between sponge withdrawal and estrus beginning was 27.7 h and 35.9 h for eCG and FSH, respectively. Interval between sponge withdrawal and the end of estrus was 55.8 h for eCG treatment and 55.6 h for FSH treatment. Estrus length was 29.3 h and 19.6 h, for eCG and FSH treatments, respectively. The biggest follicle and the second in size measured 0.74 cm and 0.54 cm for eCG treatment, whereas for the FSH treatment they measured 0.73 and 0.50 cm. The interval between the beginning of estrus and ovulation was similar within all groups: 21.0 h for eCG treated ewes and 25.2 h for the ones treated with FSH. Ewes treated with eCG presented an interval of 47.5 h between sponge withdrawal and ovulation, while the ones treated with FSH presented a 61.1 h interval. Ovulation occurred 8.3 h before the end of estrus in the eCG group. On the other hand, ewes treated with FSH ovulated 5.5 h after the end of estrus. Estrus and ovulation were efficiently synchronized in Santa Inês ewes by using long-term progestogen protocol associated to the administration of 20 mg FSH, along with Prostaglandin F2α (PGF2α) at the moment of sponge withdrawal, thus substituting the use of eCG.