Effect of hydrogen peroxide on thawed ovine sperm motility

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

Preservation of wild feline semen by freeze-drying: experimental model

According to the Convention on International Trade in Endangered Species, 36 wild feline species are threatened by extinction or severely endangered, and to save them is the target of several conservation programs. This study aimed to assess the viability of the freeze-drying technique for domestic cat sperm cells, with the ultimate goal of transferring this technology to the wild feline species. The domestic cat is an excellent experimental model for wild felids. It is in this scenario that the freeze-drying process (low-temperature vacuum dehydration) of sperm cells shows its value in preserving male cats' germplasm. Results from membrane and DNA integrity analysis are promising and validates the use of frozen-dried sperm samples in intracytoplasmic sperm injections (ICSIs). Further studies are still necessary to evaluate the ICSI embryo production using domestic cat frozen-dried sperm and the possibility of using such technology with wild felines.

Principais marcadores utilizados na avaliação da qualidade do sêmen de animais domésticos

Methods of semen cryopreservation allow changes in spermatic cells, such as damage in plasma and acrossomal membrane and modifications in mitochondrial function due to a disorder in the lipidic bilayer. For effective oocyte fertilization, spermatozoa require functional competent membranes, and intact organelles, acrosome and DNA. However, most laboratory methods used to evaluate semen quality are not highly correlated with fertilizing capacity. The discovery of a variety of fluorochromes and compounds conjugated to fluorescent probes has enabled an accurate assessment of the viability, integrity and function of spermatozoa. Among the most used probes that label the various compartments of the sperm cell there are the membrane impermeable fluorescent dyes to test the membrane integrity, as well as acylated dyes that pass the intact membrane. For the acrossomal integrity the most commonly used method is lectins labeled by a fluorescent probe. The acrosome reaction and spermatic capacitation is detected by the evaluation of membrane architecture and disorder of lipids in plasma membrane. Mitochondrial function can be determined using markers for their aerobic activity. The DNA status of spermatozoa has been determined using the metachromatic properties of Acridine Orange, and the DNA fragmentation can also be assessed by TUNEL assay. Finally, DNA condensation is analyzed using a single cell DNA gel electrophoresis assay that indicates DNA compactation. This monograph aims to compile the various tests used to detect damaged spermatozoa under cryopreservation methods, searching for improve the predictive value of semen analysis with the intention of a successful conception

Congelação de células espermáticas provenientes de epidídimo de gatos domésticos contendo antioxidante no meio diluidor

Pós-graduação em Medicina Veterinária - FCAV

Viability of Lactobacillus acidophilus La-5 in the presence of Williopsis saturnus var. suaveolens

Maintaining the viability of probiotic microorganisms from production to consumption has long been a technological challenge for the food industry. The objectives of this study were to evaluate the in vitro interaction between Lactobacillus acidophilus La-5 and Williopsis saturnus var. suaveolens and the effect of this yeast on acidification kinetics, viability of Lactobacillus acidophilus and post-acidification in fermented milk during refrigerated storage at 5 °C. The in vitro study showed a positive interaction between the acid cell free-supernatant (CFS) of probiotic bacteria La-5 and the yeast. The addition of W. saturnus var. suaveolens increased the fermentation time due to consumption of the organic acids produced by L. acidophilus. During the refrigerated storage of the samples, the presence of the yeast increased the viability of L. acidophilus and reduced post-acidification. However, the mechanism of such interaction of bacteria and yeast is not fully understood.

Immediate and late analysis of dental pulp stem cells viability after indirect exposition to alternative in-office bleaching strategies

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

Effect of LPS treatment on the viability and chemokine synthesis by epithelial cells and gingival fibroblasts

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

Impaired protamination and sperm DNA damage in a Nellore bull with high percentages of morphological sperm defects in comparison to normospermic bulls

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.

Topics in the Routine Assessment of Newborn Puppy Viability

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

Control Methods and Evaluation of Bacterial Growth on Fresh and Cooled Stallion Semen

The penis and prepuce of the stallion have a high bacterial load on its surface, forming a natural microbial flora that contaminates the semen during ejaculation. Bacterial growth in semen may cause a decline on sperm quality, viability, and fertility and predisposes the occurrence of endometritis in inseminated mares. Thus, the aim of this study was to evaluate the effect of penile wash before semen collection, the addition of different commercial skim milk-based extenders containing antibiotics (BotuSemen and INRA96), and the removal of seminal plasma by filtration on the quality, viability, and bacterial proliferation on fresh and cooled stallion semen. Animals that were never submitted to penile wash before semen collection tended to have lower bacterial contamination in the ejaculate. Semen samples extended in BotuSemen showed superiority in total motility, progressive motility, average path velocity, and rapid sperm and lower bacterial contamination in relation to semen samples extended in INRA96 after 24 hours of cooling. No difference was found in these parameters between the storage temperatures (5 degrees C and 15 degrees C). Furthermore, the removal of seminal plasma by filtration reduced the bacterial load in semen after cooling. In conclusion, the penile wash before semen collection tended to reduce the bacterial growth in fresh semen. The use of a semen extender with appropriate antibiotics and removal of seminal plasma by filtration were effective in reducing the bacterial contamination and preserved the quality of cooled stallion semen. (C) 2015 Elsevier Inc. All rights reserved.

Acidification profile, probiotic in vitro gastrointestinal tolerance and viability in fermented milk with fruit flours

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

Inappropriate management conditions, especially for the regressed class, are related to sperm quality in Prochilodus lineatus

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

Hormesis method for increasing oat straw with a view to viability of direct-seeding systems

In conservation agriculture, mainly under direct seeding, maintaining the vegetation ground cover is essential, since this serves as a reservoir of nutrients which are slowly released to plants by microorganisms. Some authors have sought to study increases in the amount of straw in the soil, in addition to slowing down the process of decomposition, with hormesis being one of the techniques used. This technique states that all chemical substances are both poisonous and nonpoisonous, with only the dosage determining whether they are lethal or not. This study aimed to evaluate the dry weight and agronomic characteristics of a crop of black oat subjected to hormesis. The experimental design was of randomised blocks, with 12 treatments and 4 replications, giving a total of 48 experimental lots. The treatments were: Haloxyfop-R Methyl Ester at dosages of 0.625, 1.25 and 2.50 g ha(-1); Glyphosate at dosages of 12.50, 25.00 and 50.00 g ha-1; 2,4-D dimethylamine salt at dosages of 100.00, 200.00 and 300.00 g ha(-1); Alterbane at a dosage of 500.00 g ha(-1); Salicylic acid at a dose of 100 g ha(-1); and a control. It was concluded that for the subdosages under test, the herbicides 2,4-D at medium dosage and Verdict at low dosage were shown to be the best treatments for conserving straw as ground cover under direct seeding.

Sperm production of Steidachneridion parahybae (Steindachner 1877) and the effect of hormonal induction throughout one reproductive cycle

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Absence of effects on the rat sperm quality after subacute exposure to low doses of fungicide prochloraz

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)