The Binder of SPerm 1 (BSP1) protein is involved in the fertilization and semen cryopreservation processes and is described to be both beneficial and detrimental to sperm. Previously, the relationship of BSP1 with freezability events has not been completely understood. The objective of this work was to determine the differential abundance of the forms of the BSP1 protein in cryopreserved seminal plasma of Bos taurus indicus bulls with different patterns of semen freezability using proteomics. A wide cohort of adult bulls with high genetic value from an artificial insemination center was used as donors of high quality, fresh semen. Nine bulls presenting different patterns of semen freezability were selected. Two-dimensional gel electrophoresis showed differential abundance in a group of seven protein spots in the frozen/thawed seminal plasma from the bulls, ranging from 15 to 17 kDa, with pI values from 4.6 to 5.8. Four of these spots were confirmed to be BSP1 using mass spectrometry, proteomics, biochemical, and computational analysis (Tukey's test at P < 0.05). The protein spot weighing 15.52 ± 0.53 kDa with a pI value of 5.78 ± 0.12 is highlighted by its high abundance in bulls with low semen freezability and its absence in bulls presenting high semen freezability. This is the first report showing that more than two forms of BSP1 are found in the seminal plasma of Nelore adult bulls and not all animals have a similar abundance of each BSP1 form. Different BSP1 forms may be involved in different events of fertilization and the cryopreservation process.
This study aimed at evaluating the relationship between physical and morphological semen features with the hypoosmotic swelling (HOS) test in raw semen of adult Nelore bulls classified as sound and unsound for breeding. Two hundred and six Nelore bulls aging from 3 to 10 years of age were subjected to breeding soundness examination. After physical and morphological semen examination, HOS test was done. After the breeding soundness examination, 94.2% of the bulls were classified as sound for breeding. There was no difference between the average scrotal circumference of bulls classified as sound and unsound for breeding (P>0.05), but there was difference between all semen physical and morphological aspects of bulls classified as sound and unsound for breeding (P>0.05), and no difference resulted in the mean percentage of reactive spermatozoa to HOS test results both for sound (38.4±17.9) and unsound animals (39.5±16.4; P>0.05), with no Pearson correlation between the HOS test and variables. According to these results HOS test can not be used alone to predict the reproductive potential of adult Nelore bulls.
RESUMO -Foram estudadas as relações entre a concentração de proteínas solúveis do plasma seminal, o teste hipoosmótico, a análise física e morfológica do sêmen e o desempenho reprodutivo de bodes da raça Alpina em regime de monta controlada. O experimento foi realizado durante os meses de fevereiro a abril de 2001. Foram realizadas 40 coletas de sêmen em quatro reprodutores adultos em regime de monta controlada. Em todas as coletas, além do exame físico e morfológico do sêmen, foram realizados os testes hipoosmóticos, isoosmóticos e a determinação da concentração de proteínas solúveis do plasma seminal. Foi detectada diferença entre os bodes na concentração de proteínas solúveis do plasma seminal, mas não houve diferença no teste hipoosmótico e no número de coberturas por prenhez. O teste hipoosmótico pode ser uma importante ferramenta para a avaliação de sêmen caprino, mas a concentração de proteínas solúveis do plasma seminal não pode ser utilizada como parâmetro para predizer a qualidade seminal e a fertilidade de bodes da raça Alpina utilizados em regime de monta natural. Palavra-chave: caprino, concentração de proteínas do plasma, teste hiposmóticoReproductive performance, soluble proteins of the seminal plasm and hypoosmotic test in male goats of the Alpine breed under controlled mating ABSTRACT -The relationships among soluble proteins of the seminal plasma, the hypoosmotic test, the physical and morphologic analyses of semen and the reproductive performance of male goats of the Alpine breed under controlled mating were studied using forty samples of semen from four adult male goats collected twice a week between February and April of 2001. Differences among male goats were observed for soluble protein concentration of the seminal plasma, but not for the hypoosmotic test and the pregnancy rate. These results suggest the hypoosmotic test may be used for the evaluation of goat semen, but the protein concentration of the seminal plasma can not be used as a parameter to predict the seminal quality and fertility of male goats of the Alpine breed.
RESUMO -Os objetivos neste estudo foram verificar se a própolis e o ácido ascórbico têm efeito sobre a integridade da membrana plasmática dos espermatozoides de caprinos e investigar o potencial desses antioxidantes no uso de meios diluidores de criopreservação de sêmen caprino. Foram utilizados cinco bodes adultos das raças Alpina (n = 2) e Saanen (n = 3). Após a coleta de sêmen, realizaram-se o exame físico do sêmen e morfológico dos espermatozoides, o teste supravital e o teste hiposmótico. Em seguida, o sêmen fresco foi diluído com o diluidor Bioxcell ® (controle); Bioxcell ® + 0,25% de extrato liofilizado de própolis; Bioxcell ® + 0,5% de extrato liofilizado de própolis; Bioxcell ® + 0,05% de ácido ascórbico; ou Bioxcell ® + 0,25% de ácido ascórbico. Após as diluições finais, foram avaliados a motilidade e o vigor espermático obtidos com cada diluidor e posteriormente o sêmen foi submetido a envase, resfriamento e congelamento. No sêmen fresco, os aspectos físicos e morfológicos e os resultados dos testes supravital e hiposmótico não diferiram entre os animais nem entre raças. As médias gerais de motilidade e vigor espermático e dos testes supravital e hiposmótico obtidos logo após o descongelamento e após 3 horas de teste de termorresistência diferiram entre si, de modo que o diluidor contendo ácido ascórbico e o controle foram similares e superiores àqueles contendo própolis. O ácido ascórbico mantém a integridade estrutural da membrana dos espermatozoides durante o processo de criopreservação, bem como sua viabilidade após o teste de termorresistência, e pode ser uma alternativa na composição de diluentes para criopreservação de sêmen caprino; a própolis não é eficaz na manutenção da integridade e da viabilidade espermática pós-descongelamento e é tóxica aos espermatozoides nas concentrações de 0,25 e 0,5%.Palavras-chave: ácido ascórbico, antioxidantes, caprinos, criopreservação, própolis, sêmen Use of propolis and ascorbic acid on goat semen cryopreservationABSTRACT -The objectives of this study were to verify whether propolis and ascorbic acid have an effect on plasmatic membrane integrity of goat spermatozoa and investigate the potential of these antioxidants in the use of goats spermatozoa cryopreservation extenders. Five adult boars were used of the Alpine (n = 2) and Saanen (n = 3) breeds. After semen collection, the evaluation consisted of the physical and morphological exam, live and dead cells (supravital test) and hyposmotic swelling test. Afterwards, the fresh semen was diluted with the Bioxcell® extender (control); Bioxcell® + 0.25% freeze dried propolis extract); Bioxcell® + 0.5% freeze dried propolis extract); Bioxcell® + 0.05% ascorbic acid) or Bioxcell® + 0.25% ascorbic acid). After final dilutions, the sperm motility and vigor were assessed obtained in each extender, and then the semen was sealed, cooled and frozen. In fresh semen, the physical and morphological aspects and the results of the supravital test and hyposmotic swelling test did not differ between animals or breeds. The gener...
-The objective of this study was to investigate the relationship between testicular echotexture and seminal quality, in addition to evaluate the testicular parenchyma echogenicity pattern in young Montana bulls classified as sound and unsound for breeding. Fifty-two young Montana Tropical Compound bulls from 22 to 33 months of age were used. The animals were previously evaluated by breeding soundness evaluation and thereafter divided into two groups of breeding soundness classes: 1 = 16 animals sound for breeding; 2 = 36 animals unsound for breeding by means of physical and morphological analysis of semen. All animals underwent an ultrasound examination of the testes, and the images were analyzed with the software "Image J". ANOVA was used for statistical analysis, to determine the effect of groups in relation to testicular measurements, physical and morphological semen features and ultrasound pixel analysis. There was a difference between scrotal circumference between classes, with 39.7±2.1 cm and 37.3±3.1 cm for sound and unsound animals, respectively. Regarding testicular echotexture, mean values for the pixel intensity were 95.7 and 94.0 for sound and unsound animals for breeding, respectively, with no difference between the classes. None of the studied parameters were correlated with each other, indicating that the quantification of the pixel intensity in testicular echotexture was not effective in determining the degree of sexual maturity in Montana Tropical Compound bulls.
The large consumption of biodegradable films from cassava starch acetate (FCSA) as ingredients in food and pharmaceutical products requires the assessment of the possible toxicity of these products. The aim of this study was to investigate the toxicity of biodegradable film from cassava starch acetate after oral exposure of Wistar rats for 90 days. The amount of food consumed and the body weight were weekly monitored. Blood and urine samples were obtained for the assessment of serum parameters and renal function. Histopathological analyses in target organs were also performed. No evidence of clinical toxicity in hematological, biochemical, or renal parameters in the FCSA-treated animals was found. In addition, relative organ weight and histopathological evaluations did not differ between groups treated with FCSA and control. Data obtained suggest that the subchronic exposure to FCSA does not cause obvious signs of toxicity in Wistar rats, indicating possible safety of this biofilm.
RESUMO IntroduçãoO teste hiposmótico avalia a atividade bioquímica da membrana plasmática intacta, devido ao influxo de água para o interior das células. Vários autores têm observado em sêmen in natura e congelado/descongelado que a integridade funcional da membrana plasmática avaliada pelo teste hiposmótico é uma importante ferramenta complementar para predizer a capacidade de fertilização do sêmen de bovinos (Bacinoglu et al., 2008). No entanto, não se entende completamente os resultados obtidos no teste hiposmótico, como também não existe padronização do protocolo para a realização da técnica em relação ao tempo de incubação e à osmolaridade da solução para sêmen de bovinos.
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