Sperm fertility and viability following 48 h of refrigeration: Evaluation of different extenders for the preservation of bull semen in liquid state

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

Adding Motility Stimulants to Improve Freezing of Buffalo Sperm Recovered from Epididymal Cauda

The cryopreservation of epididymal sperm is important to preserve genetic material from valuable buffalo bulls. This study evaluated the viability of post-thawed sperm samples recovered from the epididymal cauda adding motility inductors. For that, were used epididymides from eight Murrah buffaloes with 18 months of age. Semen samples were submitted to three different conditions: (CT - control) without adding medium, (SPERM) adding Sperm Talp medium, and (FERT) adding Fert Talp medium. Immediately after slaughter, both testicles from each animal were collected and transported at 4 degrees C at maximum six hours interval. In laboratory, the removed epididymides was flushed to obtain sperm and diluted in the freezing extender. Each buffalo sperm were divided and fractions were submitted to all conditions (CT, SPERM and FERT). Semen doses were frozen at -196 degrees C. CT, SPERM and FERT post-thawing results were 13.63 +/- 8.91, 38.77 +/- 8.91 and 42.83 +/- 8.91 for total motility, 7.30 +/- 8.74, 24.87 +/- 8.74 and 29.70 +/- 8.74 for progressive motility, 6.04 +/- 0.92, 6.74 +/- 0.92 and 6.93 +/- 0.92 for percentage of rapid cells (P < 0.05). In conclusion, diluted semen supplementation with Sperm or Fert talp increases the motility of cauda epididymal sperm of buffalo bulls.

Effect of glycerol on the viability and fertility of cooled bovine semen

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

Adaptations to salinity and osmoregulation in the frog Thoropa-miliaris (Amphibia, Leptodactylidae)

The leptodactylid frog Thoropa miliaris, from SE Brazil, may live on rocky marine shore, where it thrives on terrestrial and marine invertebrates, and often moves into the intertidal zone. The osmotic concentration of plasma, muscle homogenate and urine of the frogs freshly captured on the rocky shore was higher than those collected far from the sea, or kept in captivity for 6-8 months on a diet free of marine items. Increase in body urea and sodium concentrations, reported in amphibians as a response to hyperosmotic environment, was not found in T. miliaris. Osmotic concentration of the frogs from rocky shore was variable though, ranging from 400 to 980 mOsm/l. Such variation in the osmotic concentration may reflect a territorial behavior for foraging sites, which would result in higher intake of marine items by individuals living closer to intertidal zone.

Efeito da adição de butanol na biodegradabilidade da gasolina e do óleo diesel

The increase in the oil price and the current trend of using renewable raw materials for the production of chemicals renew the interest in the production of biobutanol that, produced by fermentation of agricultural raw materials, can be used as a component of gasoline and diesel. With the commercialization of new fuels, environmental damages due to spills can occur. Among other techniques, the clean-up of these contaminated areas can be achieved with bioremediation, a technique based on the action of microorganisms, which has the advantage of turning hazardous contaminants into non toxic substances such as CO2, water and biomass. Thus, bearing in mind the use of biobutanol in the near future as a gasoline extender and due to the lack of knowledge of the effects of butanol on the biodegradation of gasoline, this work aimed to assess the aerobic biodegradation of butanol/gasoline blends and butanol/diesel (20% v/v), being the latter compared to the ethanol/gasoline blend and biodiesel/diesel (20% v/v), respectively. Two experimental techniques were employed, namely the respirometric method and the redox indicator 2,6-dichlorophenol indophenol (DCPIP) test. In the former, experiments simulating the contamination of natural environments were carried out in biometer flasks, used to measure the microbial CO2 production. The DCPIP test assessed the capability of four inocula to biodegrade the fuel blends. In butanol/gasoline experiments the addition of the alcohols to the gasoline resulted in positive synergic effects on the biodegradation of the fuels in soil and...(Complete abstract click electronic access below)

O trabalho rural e a sazonalidade do café: um estudo sobre assalariados do município de Campo Belo - MG

Pós-graduação em Serviço Social - FCHS

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.

Semen characteristics and refrigeration in free-ranging giant anteaters (Myrmecophaga tridactyla)

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

Different extenders in the cryopreservation of bovine epididymal spermatozoa

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

N-acetilcisteína e trolox na conservação de células espermáticas epididimárias de cães

Pós-graduação em Medicina Veterinária - FCAV

Utilização de sistemas de refrigeração de sêmen equino na estabilização das amostras seminais previamente à congelação

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Criopreservación de semen bovino de la cola del epididimo utilizando los métodos convencional y automatizado

Cryopreservation of sperm is important to preserve the germplasm from animals of genetic value, which can die unexpectedly. This study compares conventional and automated methods of cryopreservation of spermatozoa obtained from the epididymis of bulls post-mortem. Twenty-two epididymides were obtained from a commercial slaughterhouse. Spermatozoa were collected from the tail of the epididymis using the retrograde flow technique. Thus, the samples, which were diluted in 10 ml of extender without glycerol (Botubov® I, Botupharma, Botucatu, SP, Brazil), were evaluated on motility, sperm vigor, structural and functional (swelling hypoosmotic test) membrane integrity, mitochondrial activity, sperm viability and ADN fragmentation. The samples were divided into two aliquots and diluted in extender with glycerol (Botubov® II, Botupharma, Botucatu, SP, Brazil) at a concentration of 50x106 motile sperm/0.5 French straws. One sample was frozen by the conventional method (4 hours at 5°C, in a refrigerator and 20 min in nitrogen vapor) and the other by the automated method (Cryogen® Dualflex, Neovet, Uberaba, MG, Brazil). The parameters were higher in all the tests of fresh sperm samples, with the exception of the swelling hypoosmotic test, which showed no significant difference when the results were compared with sperm frozen by the conventional method. The average motility of fresh spermatozoa was 74%, and conventional and automated averages were 29 and 25%, respectively. Therefore, although cryopreservation techniques reduce sperm quality parameters, the viability of the sperm is maintained, and these methods can be used to preserve sperm.

Comparison of apoptotic cells between cryopreserved ejaculated sperm and epididymal sperm in stallions

The development of a reliable technique to freeze epididymal semen would provide a unique opportunity to preserve valuable genetic material from unexpectedly lost stallions. The aim of this study was to compare the apoptotic indices of sperm obtained from ejaculate, sperm recently recovered from the epididymides (EP), and sperm recovered from epididymides stored at 5 C for 24 hours (EP-stored). For the first category, two ejaculates from seven stallions were collected and then submitted to cryopreservation using an egg yolk-based extender. One week after the last semen collection, the stallions were submitted to bilateral orchiectomy, and sperm from one of the cauda epididymis was harvested immediately after castration (EP). The remaining testicle was stored in a passive refrigeration container at 5 C for 24 hours before the cauda epididymal sperm was harvested (EP-stored). Sperm harvesting from the epididymis for EP and EP-stored was performed by retrograde flushing of the caudal portion of the epididymis using a skim milk-based extender. The recovered sperm was then cryopreserved using the egg yolk-based extender. Sperm motility parameters were studied by computerassisted semen analysis, and apoptosis was estimated by measuring caspase activity and membrane phospholipid translocation using epifluorescence microscopy. The samples were evaluated immediately (0 hour) and 8 hours after thawing. At 0 hour, no differences in sperm parameters were observed among the groups, but after 8 hours, significant statistical differences were observed in sperm motility parameters and plasma membrane integrity among the treatment groups. In addition, viable cells with no apoptotic signs were more prevalent in EP and EP-stored, suggesting that epididymal sperm is less sensitive to the cold shock caused by sperm cryopreservation.

The methylation patterns of the IGF2 and IGF2R genes in bovine spermatozoa are not affected by flow-cytometric sex sorting

The objectives of this study were to investigate the effect of sexing by flow cytometry on the methylation patterns of the IGF2 and IGF2R genes. Frozen-thawed, unsorted, and sex-sorted sperm samples from four Nellore bulls were used. Each ejaculate was separated into three fractions: non-sexed (NS), sexed for X-sperm (SX), and sexed for Y-sperm (SY). Sperm were isolated from the extender, cryoprotectant, and other cell types by centrifugation on a 40:70% Percoll gradient, and sperm pellets were used for genomic DNA isolation. DNA was used for analyses of the methylation patterns by bisulfite sequencing. Methylation status of the IGF2 and IGF2R genes were evaluated by sequencing 195 and 147 individual clones, respectively. No global differences in DNA methylation were found between NS, SX, and SY groups for the IGF2 (P=0.09) or IGF2R genes (P=0.38). Very specific methylation patterns were observed in the 25th and 26th CpG sites in the IGF2R gene. representing higher methylation in NS than in the SX and SY groups compared with the other CpG sites. Further, individual variation in methylation patterns was found among bulls. In conclusion, the sex-sorting procedure by flow cytometry did not affect the overall DNA methylation patterns of the IGF2 and IGF2R genes, although individual variation in their methylation patterns among bulls was observed. Mol. Reprod. Dev. 79:7784, 2012. (C) 2011 Wiley Periodicals, Inc.

Chain extension of poly (ethylene terephthalate) by reactive extrusion with secondary stabilizer

Poly(ethylene tereftalate) (PET) is a polymer highly susceptible to the hydrolytic reactions that occur during applications and mainly in thermomechanical processing. These reactions lead to the decrease of molecular weight of the polymer, limiting the recycling number of the material. The reactive extrusion of the PET in presence of chain extenders is an alternative to recover mechanical and rheological properties that were depreciated by the polymer degradation. In this study, PET wastes from nonwoven fabrics production were extruded in presence of the secondary stabilizer Irgafos 126 (IRG) on variable concentrations. The results showed that Irgafos 126 increased molecular weight, decreased crystallinity and changed processing behavior of the PET, similarly to the effects produced by the well-known chain extender pyromellitic dianhydride (PMDA), showing that the secondary stabilizer Irgafos 126 can also act as a chain extender for the PET.