Sperm motility parameters for Steindachneridion parahybae based on open-source software

Summary: The objective of this work was to evaluate the sperm motility of 13 Steindachneridion parahybae males using open-source software (ImageJ/CASA plugin). The sperm activation procedure and image capture were initiated after semen collection. Four experimental phases were defined from the videos captured of each male as follows: (i) standardization of a dialogue box generated by the CASA plugin within ImageJ; (ii) frame numbers used to perform the analysis; (iii) post-activation motility between 10 and 20 s with analysis at each 1 s; and (iv) post-activation motility between 10 and 50 s with analysis at each 10 s. The settings used in the CASA dialogue box were satisfactory, and the results were consistent. These analyses should be performed using 50 frames immediately after sperm activation because spermatozoa quickly lose their vigor. At 10 s post-activation, 89.1% motile sperm was observed with 107.2 μm s-1 curvilinear velocity, 83.6 μm s-1 average path velocity, 77.1 μm s-1 straight line velocity; 91.6% were of straightness and 77.1% of wobble. The CASA plugin within ImageJ can be applied in sperm analysis of the study species by using the established settings. © 2013 Blackwell Verlag GmbH.

Genetic parameters for scrotal circumference, breeding soundness examination and sperm defects in young Nellore bulls

A total of 51,161 records of scrotal circumference measurements at 18 mo of age (SCI 8) and 17,648 records of sperm defects and breeding soundness of Nellore bulls (mean age of 22.5 mo), raised under extensive conditions, were analyzed to estimate coefficients of heritability and genetic correlations of morphological semen traits by Bayesian inference. The observed semen traits were classified as minor (MID). major (MAD), and total sperm defects (TD). The animals were classified according to breeding soundness as satisfactory and unsatisfactory potential breeders. The (co)variance components and breeding values were estimated by Gibbs sampling using the GIBBS2F90 program under an animal model that included contemporary group as fixed effect, age of animal as linear covariate, and direct additive genetic effects as random effects. Heritabilities of 0.40 ± 0.02, 0.16 ± 0.02, 0.04 ± 0.01, 0.15 ± 0.01, and 0.10 ± 0.01 were obtained for SCI8, MID, MAD, TD, and breeding soundness, respectively. The SC18 showed a positive and moderate correlation with breeding soundness (0.56 ± 0.04) and a negative and low correlation with MID (-0.23 ± 0.03), MAD (-0.16 ± 0.02), and TD (-0.24 ± 0.02). In conclusion, scrotal circumference showed the best response to selection among the traits studied and was favorably correlated with breeding soundness and sperm morphology in young Nellore bulls. © 2013 American Society of Animal Science. All rights reserved.

Identification of phospholipase C zeta in normospermic and teratospermic domestic cat sperm

In mammalian species, oocyte activation is initiated by oscillations in the intracellular concentration of free calcium ([Ca2+]i), which are also essential to allow embryonic development. To date, evidence supporting the hypothesis that a sperm factor is responsible for initiating oocyte activation has been presented in various mammalian species. Among the possible candidates to be the active sperm factor is the novel sperm-specific phospholipase C ζ (PLCζ), which besides its testis-specific expression is capable of initiating [Ca2+]i oscillations. In this study, we investigated the presence of PLCζ in the sperm of the domestic cat and whether normospermic and teratospermic cats differ in their PLCζ expression. Immunoblotting with anti-PLCζ antibodies confirmed the presence of an immunoreactive band of ~70 kDa in whole sperm lysates of domestic cat as well as in both soluble and insoluble fractions from this sperm. Additional immunoreactive bands, probably C- and N-terminal truncated versions of PLCζ, were also visualized in the soluble sperm fractions. Interestingly, immunoreactivity of PLCζ was detectable in teratospermic sperm, although with slightly less intensity than in normospermic sperm. In conclusion, domestic cat sperm express PLCζ in both cytosolic and high-pH fractions, which is consistent with data in other mammals. Sperm from teratospermic cats also express PLCζ, albeit at reduced concentrations, which may affect the fertility of these males. © 2013 Elsevier Inc..

Cryoprotective effect of different glycerol concentrations on domestic cat spermatozoa

Cryopreservation of spermatozoa is a pivotal tool in assisted reproduction, and studies aiming to establish optimal freezing/thawing protocols are essential to enhance sperm survival. The objectives of the present study were to (1) compare the cryoprotective efficiency of three different glycerol concentrations (3%, 5%, and 7%) on the basis of post-thaw sperm quality and (2) investigate whether the incidence of morphologically abnormal sperm in fresh samples is related to cryodamage sensitivity. Semen was collected from six tomcats using an artificial vagina (total 18 ejaculates). Each ejaculate was diluted using Tris-egg yolk-based extender (TEY), evaluated, equally divided into three aliquots, and rediluted using TEY with and without glycerol to achieve final concentrations of 3%, 5%, and 7%. Samples were loaded into 0.25 mL straws, equilibrated for 60 minutes at 5 °C, frozen, and then thawed at 46 °C for 12 seconds. Fresh and frozen-thawed samples were evaluated for sperm motion parameters (computer-assisted sperm analysis), plasma membrane integrity (PMI; propidium iodide and carboxyfluorescein diacetate), and DNA integrity (acridine orange). Plasma and acrosomal membrane integrity were assessed by flow cytometry (propidium iodide and fluorescein isothiocyanate-conjugated pea (Pisum sativum) agglutinin) immediately after thawing. Sperm motion parameters were also evaluated at 30 and 60 minutes of postincubation. For all treatment groups, cryopreservation significantly impaired the PMI and sperm motion parameters, except for straightness and amplitude of lateral head displacement. DNA integrity showed a slight reduction (P < 0.05) when 3% glycerol was used. The percentage of total motility, progressive motility, and rapid spermatozoa were significantly lower immediately after thawing and up to 60 minutes of incubation for the 3% glycerol group when compared with 5% and 7%. No difference (P > 0.05) was found for PMI, acrosome integrity, and DNA integrity among post-thaw groups. However, higher (P < 0.05) incidence of viable cells with reacted acrosome and dead cells with intact acrosome were observed with 7% and 3% glycerol, respectively. Percentage of morphologically abnormal spermatozoa in fresh sample was positively correlated with PMI only in the 3% glycerol group and negatively correlated with sperm motility in the 5% and 7% groups. In conclusion, the final concentration of 5% glycerol offered better cryoprotective effect for ejaculated cat sperm, and the relationship found between prefreezing sperm morphology and post-thaw sperm quality showed to be dependent on final glycerol concentration. © 2013 Elsevier Inc.

Uso de antioxidantes no meio diluente tris para refrigeração e congelação de células espermáticas provenientes de epidídimo de cães submetidos à orquiectomia eletiva

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Comparação entre diferentes taxas de sêmen na viabilidade espermática em garanhões de alta e baixa congelabilidade

Pós-graduação em Medicina Veterinária - FMVZ

Efeito do meio e da dose inseminação sobre a congelabilidade e fertilidade de espermatozóides recuperados da cauda do epidídimo de garanhões

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Congelação e efeito do meio sobre as características físico-químicas do sêmen suíno no inverno e verão

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influência dos danos oxidativos na qualidade espermática em bovinos de diferentes idades

Pós-graduação em Medicina Veterinária - FCAV

Efeito da adição da asolctina e fosfatidilcolina de soja em meio à base de gema de ovo sobre os parãmetros espermáticos e fertilidade de sêmen congelado de garanhões

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efeitos do processo de seleção do sexo por centrifugação em gradiente de densidade na qualidade de espermatozódes bovinos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efeito do armazenamento por 24 horas em diferentes sistemas de refrigeração sobre a viabilidade e fertilidade de sêmen congelado eqüino

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Criopreservação de sêmen do epididimo de gatos domésticos (felis catus) após refrigeração por 24 horas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeito do meio de congelação sobre as características morfofuncionais do sêmen canino

Pós-graduação em Medicina Veterinária - FMVZ

Estudo de metodologias para as criopreservação de sêmen de jumento(Equus Asinus) por meio de testes laboratoriais e fertilidade

Pós-graduação em Medicina Veterinária - FMVZ