Eneder Rosana Oberst

Two-dimensional polyacrylamide gel electrophoresis of bovine seminal plasma proteins and their relation with semen freezability

The objective of this study was to evaluate the low weight (10-30 kDa) protein profile of bovine seminal plasma using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and to determine if any of these proteins was associated with semen freezability. Seminal plasma was collected from 16 bulls of high or low semen freezability. Twelve protein spots were identified from the 2D gel (15%); six of these were present in all samples. Of the 12 proteins found, three spots, present in all samples, 3 (15-16 kDa), 5 (16-17 kDa), and 7 (10-12 kDa) had nonsignificant variation among bulls, regardless of their freezability classification. Four proteins were more abundant (P<0.05) in seminal plasma samples collected from bulls with high semen freezability than in samples of bulls with low semen freezability: the spots 3 (15-16 kDa, pI 4.7-5.2), 7 (11-12 kDa, pI 4.8-4.9), 11 (13-14 kDa, pI 4.0-4.5), and 23 (20-22 kDa, pI 4.8-5.2). On the other hand, spot 25 (25-26 kDa, pI 6.0-6.5) was more abundant (P<0.05) on seminal plasma samples from bulls with low semen freezability. The N-terminus sequence of protein 7 was identical to the acidic seminal fluid protein (aSFP). Protein 23 (after trypsin digestion) had structural similarity to bovine clusterin. We concluded that there were differences in the seminal plasma protein profile from bulls with low and high semen freezability; aSFP, clusterin, proteins 3 and 11 may be used as semen freezability markers; and protein 25 was related to low semen freezability.

Criopreservação de espermatozóides eqüinos comparando duas curvas de congelamento combinadas com diluentes comerciais: uma análise laboratorial

During semen cryopreservation, sperm cells were submitted to several deleterious events leading to membrane damage which result in fertility decrease. This study was designed to compare the effects of two freezing techniques (conventional and automated), and the use of two commercial extenders as cryoprotectants (FR-5® and Botu-Crio®) on total and progressive motility, integrity and functionality of spermatic membranes during the cryopreservation of equine semen. Twenty ejaculates from two stallions were analyzed. The total and progressive motility of fresh and post-thawing semen samples were evaluated by patterns assays. Function of plasmatic membrane was measured by the hipoosmotic swelling test. Integrity of plasmatic membrane was evaluated using carboxifluorescein diacatate and iodidium propide fluorescent probes. There were significant differences between the two freezing techniques and/or between cryoprotectants for all assessed parameters. The combination of Botu-Crio® and automated curves showed better results on total and progressive post-thawing motility. The extender Botu-Crio®, alone, showed to better preserve the membrane integrity and function.

Variação estacional na qualidade do sêmen de carneiros Lacaune no Brasil

Existem poucos dados sobre a producao de semen da raca Lacaune, no sul do pais. Assim, o objetivo deste estudo foi o de avaliar a producao e a qualidade do semen de carneiros desta raca, criados no estado do Rio Grande do Sul (RS). Foram utilizados 12 carneiros, de idades entre 1-4 anos, criados em condicoes intensivas. O semen foi colhido por vagina artificial e foram avaliadas as caracteristicas quantitativas e qualitativas e a circunferencia escrotal, do inverno de 2002 ate a primavera de 2003. Com excecao da concentracao espermatica, foram observadas variacoes estacionais em todas as caracteristicas estudadas. O volume variou de 1.1 ± 0.4 mL, no inverno de 2002, a 1.5 ± 0.4 mL no outono de 2003. O numero total de espermatozoides por ejaculado variou de 4.0 ± 3.3x10(9) (inverno de 2002) a 5.7 ± 3.3x10(9) (verao de 2003). Observou-se variacao estacional significativa (P < 0,05) na circunferencia escrotal em 2002, bem como variacao significativa entre as idades dos animais, no inverno do mesmo ano. Os valores medios da producao espermatica observados no verao e o outono foram superiores aos obtidos no inverno e primavera. O verao e outono foram considerados as estacoes mais indicadas para a utilizacao de machos da raca Lacaune em programas reprodutivos, no RS. Mais estudos devem ser conduzidos, para a verificacao de efeitos estacionais sobre o congelamento do semen.

Functional integrity of Colossoma macropomum (Cuvier, 1816) sperm cryopreserved with enriched extender solutions

Cryoprotectant solutions are used to protect the sperm from alterations caused by the low temperature in the cryopreservation process. We evaluated the quality of Colossoma macropomum semen after freezing, using dimethyl sulfoxide (DMSO) as a cryoprotectant, combined with two extender solutions (T1 - Solution 1: Glucose 90.0 g/L, Sodium Citrate 6.0 g/L, EDTA 1.5 g/L, Sodium Bicarbonate 1.5 g/L, Potassium Chloride 0.8 g/L, Gentamycin Sulphate 0.2 g/L, and T2 - Solution 2: Glucose 90.0 g/L, ACP(r)-104 10.0 g/L). Motility rate and motility time did not differ between T1 and T2 and were lower than fresh semen. The number of normal sperm was significantly different in treatments T1 (15.1%) and T2 (21.9%), and both showed a reduction in the percentage of normal sperm compared to fresh semen (57.4%). The values found for the rates of fertilization and hatching, mitochondrial functionality and sperm DNA, did not differ between the treatments (T1 and T2). Regarding membrane integrity, there was a higher percentage of spermatozoa with intact membranes in T1 (53.4%) than T2 (43.7%). The extender solutions, combined with 10% DMSO, maintained the sperm DNA intact in almost all the C. macropomum sperm cells, however there was a loss in their functionality.