Assessment of different sperm functional tests in golden-headed lion tamarins (Leontopithecus chrysomelas).

The golden-headed lion tamarin (Leontopithecus chrysomelas) is an endangered species endemic to Brazil's Atlantic Forest, a shrinking biodiversity hotspot. As in other Neotropical primates, its semen characteristics and freezability are poorly studied. Hence, reproductive technologies for callitrichids would greatly benefit from reliable methods of semen analysis. In a bid to promote reproductive research in tamarins, we validated simple and inexpensive sperm function tests that can be used to monitor sperm-egg binding, plasma membrane and acrosome integrity, mitochondrial activity, and DNA fragmentation. Ejaculates from adult males were individually diluted and divided into control and damage-induced aliquots, and then samples comprising assorted amounts of damaged spermatozoa were examined by organelle-specific tests. Our findings showed that sperm-binding in chicken egg perivitelline membrane (EPM) positively correlated with the number of spermatozoa injured by snap-freezing. Eosin-nigrosin (EN) and propidium iodide readings were correlated with each other, and both provided robust measurements of plasma membrane integrity. A high correlation between expected and measured amounts of acrosome-intact spermatozoa was found using Fast Green-Rose Bengal (FG-RB), Coomassie Blue (CB), and FITC-PSA stains, and all three methods exhibited comparable results. Likewise, different percentages of UV-irradiated spermatozoa were accurately assessed for DNA integrity by Toluidine Blue (TB) and sperm chromatin dispersion (SCD) tests. Comparisons between 3,3'-diaminobenzidine (DAB) and JC-1 stains also indicated the reliability of the former assay to ascertain gradual increases in spermatozoa with greater mitochondrial function. These data confirmed that different parts of the tamarin spermatozoa can be simply and consistently evaluated by EPM, EN, FG-RB, CB, TB, and DAB protocols.

Influence of european quail breeders age on egg quality, incubation, fertility and progeny performance

The influence of age (85, 140, and 270 days) of European quails breeders on the egg quality and hatching, fertility and progeny performance was evaluated. The experimental design was completely randomized in a 3x3 factorial arrangement (females' age x males' age), with ten replicates and six birds per experimental unit (four females and two males). Egg production and quality were determined during 3 periods of 14 days and incubation parameters were evaluated in eggs obtained in five consecutive days. The live performance of the progenies was analyzed until 35 days. There was no effect of male age or any interaction between the age of males and females for the evaluated variable. The female's age influenced egg production, egg weight and chick weight, with better results obtained for 140-d-old breeders. The age of females reduced the hatchability, increased the late mortality in incubated eggs, and had no effect on fertility, total embryo mortality or eggshell structure, when analyzed by electron microscopy. The number of sperm trapped in the outer perivitelline layer (sptz/mm²) was determined in 10 fertile eggs per experimental unit. Young females fertilized by young males (80 days) had reduced sptz/mm². Progeny live performance was not affected by breeder's age. Breeders with 270 days retain fertility; however, their egg production, weight and hatchability of fertilized eggs is reduced. In conclusion, European quail breeders with 140 days of age have better egg quality, hatching and breeding results.

Influence of european quail breeders age on egg quality, incubation, fertility and progeny performance

The influence of age (85, 140, and 270 days) of European quails breeders on the egg quality and hatching, fertility and progeny performance was evaluated. The experimental design was completely randomized in a 3x3 factorial arrangement (females' age x males' age), with ten replicates and six birds per experimental unit (four females and two males). Egg production and quality were determined during 3 periods of 14 days and incubation parameters were evaluated in eggs obtained in five consecutive days. The live performance of the progenies was analyzed until 35 days. There was no effect of male age or any interaction between the age of males and females for the evaluated variable. The female's age influenced egg production, egg weight and chick weight, with better results obtained for 140-d-old breeders. The age of females reduced the hatchability, increased the late mortality in incubated eggs, and had no effect on fertility, total embryo mortality or eggshell structure, when analyzed by electron microscopy. The number of sperm trapped in the outer perivitelline layer (sptz/mm²) was determined in 10 fertile eggs per experimental unit. Young females fertilized by young males (80 days) had reduced sptz/mm². Progeny live performance was not affected by breeder's age. Breeders with 270 days retain fertility; however, their egg production, weight and hatchability of fertilized eggs is reduced. In conclusion, European quail breeders with 140 days of age have better egg quality, hatching and breeding results.(AU)

In vitro fertilization of the rock shrimp, Rhynchocinetes typus (Decapoda, Caridea): a review

This review compiles all the research done on gametes and fertilization in the rock shrimp, R. typus, and describes the sequence of events from the first gamete interaction to zygote formation and the first cleavage of the embryo, with light, fluorescence confocal and electron microscopes. Early studies showed that sperm from the vas deferens have a tack-shape with a "needle-like process" or rigid spike (RS) that extends from a semi-spherical body that contains the arms with chromatin and spines. Upon contact with seawater and by action of Na +, the arms and spines extend, producing an inverted umbrella form of the spermatozoa. The first sperm-oocyte interaction occurs between protein receptors type lectins of the sperm RS and oocyte chorion sperm ligands. These ligands contain residues of a-Glu, Man (a 1-3) Man, a and p-GlcNAc and a-GalNA terminal residues. It was found that a-Man and GlcNAc residues are the ligands that are directly related to the adhesion process and further penetration of sperm. After this first interaction, the RS enters the oocyte envelope by the action of a trypsin-like enzyme, rhynchocinecine, present in the acicular process. Later, arms and spines penetrate the oocyte cytoplasm, where the chromatin of the arms begin to migrate to the central area of the sperm, condensing in a cup-shaped structure near the connecting piece, which forms the male pronucleus.

A carbohydrate-based mechanism of species recognition in sea urchin fertilization

In the present review, we describe a systematic study of the sulfated polysaccharides from marine invertebrates, which led to the discovery of a carbohydrate-based mechanism of sperm-egg recognition during sea urchin fertilization. We have described unique polymers present in these organisms, especially sulfated fucose-rich compounds found in the egg jelly coat of sea urchins. The polysaccharides have simple, linear structures consisting of repeating units of oligosaccharides. They differ among the various species of sea urchins in specific patterns of sulfation and/or position of the glycosidic linkage within their repeating units. These polysaccharides show species specificity in inducing the acrosome reaction in sea urchin sperm, providing a clear-cut example of a signal transduction event regulated by sulfated polysaccharides. This distinct carbohydrate-mediated mechanism of sperm-egg recognition coexists with the bindin-protein system. Possibly, the genes involved in the biosynthesis of these sulfated fucans did not evolve in concordance with evolutionary distance but underwent a dramatic change near the tip of the Strongylocentrotid tree. Overall, we established a direct causal link between the molecular structure of a sulfated polysaccharide and a cellular physiological event - the induction of the sperm acrosome reaction in sea urchins. Small structural changes modulate an entire system of sperm-egg recognition and species-specific fertilization in sea urchins. We demonstrated that sulfated polysaccharides - in addition to their known function in cell proliferation, development, coagulation, and viral infection - mediate fertilization, and respond to evolutionary mechanisms that lead to species diversity.