RESUMODeterminou-se a concentração de proteína total e de suas frações (albumina, alfa globulina, beta

The aim of this study was to evaluate the chromatin packing and sperm head morphometry of cryopreserved semen of Nelore bulls (Bos taurus indicus) of different ages. Furthermore, the influence of the degree of chromatin compaction on in vitro embryo production (IVP) was investigated. Forty bulls were divided into three groups: young (1.8-2 years), adult (3.5-7 years), and senile (8-14.3 years). The ejaculates were frozen according to standards established by the Artificial Insemination Center located in the Southeast of Brazil. Toluidine blue staining was used for simultaneous evaluation of the sperm chromatin and sperm head morphometry. Chromomycin A3 (CMA3) was applied to analyze sperm protamination and IVP for embryonic development. Spermatozoa of young bulls presented higher values for area (A, pixels), perimeter (P, pixels), and width (W, pixels) compared to adults and senile (young: A = 1848.5 ± 119.79, P = 10.23 ± 0.29, and W = 1.95 ± 0.1; adults: A = 1672.9 ± 104.46, P = 9.86 ± 0.33, and W = 1.81 ± 0.06; senile: A = 1723.1 ± 124.41, P = 9.97 ± 0.33, and W = 1.83 ± 0.09; P < 0.0001) and showed higher protamination deficiency when analyzed by CMA3 (young: 1.57 ± 0.76; adults: 1.09 ± 0.63, and senile: 0.90 ± 0.59; P < 0.05). Likewise, variables of sperm head size (A, P, and W) and protamination assessed by CMA3 showed negative correlation with age and positive correlation with ellipticity, evaluated by toluidine blue method (P < 0.05). Sperm head area was larger in spermatozoa presenting chromatin instabilities than spermatozoa without chromatin alteration (P < 0.0001). There was no difference in IVP when using semen with larger or smaller portions of spermatozoa with chromatin instabilities, indicating that the proportion of sperm with abnormal chromatin compaction (4%-16.15%) did not interfere with early embryonic development. From our results, it can be concluded that sperm of young Nelore bulls have larger heads compared to adults and senile due to reduced protamine content when evaluated by CMA3 and higher proportion of major sperm defects assessed by differential interference contrast microscopy.

No abstract

The routine semen evaluation assessing sperm concentration, motility and morphology, does not identify subtle defects in sperm chromatin architecture. Bulls appear to have stable chromatin, with low levels of DNA fragmentation. However, the nature of fragmentation and its impact on fertility remain unclear and there are no detailed reports characterizing the DNA organization and damage in this species. The intensive genetic selection, the use of artificial insemination and in vitro embryo production associated to the cryopreservation process can contribute to the chromatin damage and highlights the importance of sperm DNA integrity for the success of these technologies. Frozen-thawed semen samples from three ejaculates from a Nellore bull showed high levels of morphological sperm abnormalities (55.8±5.1%), and were selected for complementary tests. Damage of acrosomal (76.9±8.9%) and plasma membranes (75.7±9.3%) as well as sperm DNA strand breaks (13.8±9.5%) and protamination deficiency (3.7±0.6%) were significantly higher compared to the values measured in the semen of five Nellore bulls with normospermia (24.3±3.3%; 24.5±6.1%; 0.6±0.5%; 0.4±0.6% for acrosome, plasma membrane, DNA breaks and protamine deficiency, respectively) (P<0.05). Motility and percentage of spermatozoa with low mitochondrial potential showed no differences between groups. This study shows how routine semen analyses (in this case morphology) may point to the length and complexity of sperm cell damage emphasizing the importance of sperm function testing.

Este estudo correlacionou os tempos de jejum sólido pré-anestésico com alterações nos níveis de glicemia plasmática, cortisol sérico, estado clínico e equilíbrio ácido-base em cães submetidos a anestesia geral inalatória. Utilizaram-se oito animais, adultos, sem raça definida, distribuídos de acordo com o período de jejum sólido: Grupo 1 (12 horas), Grupo 2 (18 horas) e Grupo 3 (24 horas). Foi acompanhado o esvaziamento do conteúdo gástrico e em seguida, todos animais foram submetidos ao mesmo procedimento anestésico. Freqüência cardíaca e respiratória, temperatura retal, tempo de reperfusão capilar, grau de hidratação e pressão arterial não-invasiva foram mensurados previamente à administração de acepromazina, 10 minutos decorridos da mesma e a cada 10 minutos durante a manutenção anestésica, incluindo-se ETCO2; valores hemogasométricos (pH, PaCO2, PaO2, HCO3, CO2 total, SatO2, déficit de base), glicêmicos e de cortisol sérico foram avaliados previamente à MPA e a cada trinta minutos durante a manutenção anestésica. No período de recuperação anestésica, novas dosagens glicêmicas e de cortisol foram realizadas. Constataram-se poucas alterações cardiocirculatórias e respiratórias durante a anestesia, não havendo interferência dos diferentes tempos de jejum. Os animais com 12 horas de jejum pré-anestésico apresentaram glicemia mais elevada do que os demais grupos, no período de recuperação anestésica. As concentrações de cortisol não foram afetadas pelo jejum. O jejum pré-anestésico sólido, independente do tempo de duração, caracterizou um quadro de discreta alcalose respiratória. Todos os animais apresentaram-se em bom estado clínico nos três grupos. Recomenda-se jejum pré-anestésico sólido de 18 horas para garantir ausência completa de conteúdo alimentar sólido no estômago.