Criopreservação de sêmen humano: comparação entre métodos de congelação e tipos de envase

OBJETIVOS: comparar duas diferentes técnicas de congelação e dois tipos de envase do sêmen humano durante processo de criopreservação. MÉTODOS: estudo experimental, no qual foi analisada a criopreservação de 18 amostras de sêmen de 18 voluntários. Após a adição de meio crioprotetor, "Test-yolk buffer" , as amostras de sêmen foram envasadas em palhetas com capacidade de 0,25 mL ou em criotubos de 2 mL e submetidas à criopreservação por dois métodos, um lento e outro rápido, totalizando quatro tratamentos distintos: RP (congelação pelo método rápido e envasado em palheta), RT (rápido-criotubo), LP (lento-palheta) e LT (lento-criotubo). As amostras, após 24 horas, foram descongeladas em temperatura ambiente e mantidas a 37ºC. Os dados coletados foram analisados através do teste t de Student, com p<0,05, utilizando o programa de computador SPSS for Windows® versão 11.0.0. RESULTADOS: houve redução da motilidade espermática após o processo de criopreservação. A taxa de motilidade inicial foi 58,1% e as motilidades após os diferentes métodos de criopreservação foram: 19,2% (RP), 27% (RT), 21,1% (LP) e 30,3% (LT). Houve redução significativa na morfologia normal. A taxa de morfologia normal inicial foi 14,2% e as morfologias após os diferentes métodos de criopreservação foram: 12,8% (RP), 12,6% (RT), 12,6% (LP) e 12,4% (LT). CONCLUSÕES: o método de criopreservação lento com envase em criotubo esteve associado à melhor motilidade espermática após o descongelamento. Não houve diferença entre os métodos quando avaliada a morfologia espermática.

Evaluation of Mycobacterium avium subsp. paratuberculosis faecal culture protocols and media

Paratuberculosis is an important enteritis of ruminants caused by Mycobacterium avium subsp. paratuberculosis (Map). The disease is officially considered exotic in Brazil, but recent serological surveys and the isolation of the agent suggest it may occur in our herds. The aim of this study was to evaluate three different formulations of Herrold's egg yolk agar with mycobactin J (HEYM) and four faecal culture protocols considering their ability for Map growth as well as cost and ease of application. Three formulations of HEYM were inoculated with two suspensions of Map. Spiked faeces and naturally contaminated faecal samples were treated by the four faecal culture protocols. Centrifugation protocol and HEYM recommended by OIE showed the best results on the recovery of Map.

Paratuberculosis in a dairy Gyr herd in the State of Paraíba, Brazil

This paper describes the clinical, pathological, and microbiologic aspects of paratuberculosis (Johne's disease) in a dairy Gyr herd in the State of Paraíba, northeastern Brazil. An eight years old cow with chronic unresponsive diarrhea was clinically examined and euthanized for pathological evaluation. Fecal samples from all 160 animals over 12 months of age from the herd were collected for isolation of Mycobacterium avium subsp. paratuberculosis. Clinically, the index case cow was severely dehydrated, cachectic, with profuse mucous diarrhea. The main post-mortem findings were emaciation and thickened intestinal wall. Microscopically, the intestinal lamina propria and submucosa were infiltrated by macrophages, epithelioid cells, and Langhans giant cells with numerous alcohol-acid resistant bacilli in the cytoplasm. Two fecal samples displayed growth in slants of Herrold's egg-yolk agar supplemented with mycobactin J, 150 days after incubation. No growth was noticed in slants without mycobactin J. Microscopic examination of the isolated microorganisms stained by Ziehl-Neelsen revealed considerable amounts of alcohol-acid resistant bacilli, morphologically compatible with Mycobacterium spp. Based on the clinical signs, gross and histological lesions, growth time, bacterial morphology in Ziehl-Neelsen staining, and dependence of mycobactin J, the first diagnosis of paratuberculosis in Zebu cattle was made.

Ultrasonographic and radiographic determination of egg development of jurarás (Kinosternon scorpioides) in captivity

Kinosternon scorpioides (Linnaeus, 1766), with its common name of jurará, is a quite variable species of turtles, and many different names have been applied to populations throughout its range. Currently, however, four subspecies are considered valid as K. scorpioides arises from southern Panama over most of northern South America and is found in Ecuador, northern Peru, southern Bolivia, northern Argentina, eastern Guyana and Brazil. Thus, an ultrasonographic and radiographic study was performed in order to describe the morphology and development of eggs of 20 female jurará mud turtles K. scorpioides, from September 2005 to August 2006. In the first month, the ovarian cycle was characterized by absence of vitellogenic follicles, atresic follicles or oviduct eggs. From October 2005 to March 2006 on, ultrasonographic scanning allowed to establish the growing vitellogenic follicles. Vitellogenic follicles were observed with spherical to ovoid shapes, with a surrounding echogenic yolk, a nonechogenic albumin layer, and a high echogenic shell. The oviduct eggs were identified by radiography just 180 days after beginning the experiment, when the shell became enough mineralized to impress the radiographic film. This experiment allowed to obtain by means of the 7.5 MHz linear probe images with adequate resolution and penetration for visualization of follicles. Successive ultrasonographic examinations of 20 K. scorpioides females allowed to access initial stages of vitellogenic follicles and oviduct eggs, and radiographic examination revealed to be an easy technique to assess oviduct eggs and to allow evaluation of egg development in jurarás, from 6 months on.

Character transformations and their functional significance as a key to the evolution of hystricognath Rodentia

Hystricognathi represent a monophyletic taxon within Rodentia. Since phylogenetically analyzed morphological systems are essential for revealing evolutionary processes, this study identifies evolutionary character transformations on the stem lineage of Hystricognathi as derived from the author's own work and the literature. Data so far indicate that evolutionary transformations in the rostral head region, the loss of tactile ability in the outer nasal skin and the mobile arrangement of the associated cartilage, were allied with a switch from omnivorous to herbivorous and fiber-rich nutrition. Additional character transformations in the skull assist in digesting such food. Structures associated with reproduction and placentation show a remarkable pro portion of derived character conditions: the chorioallantoic placenta has a ring-shaped organization and growth structure which optimizes the capacity for passive diffusion, a subplacenta occurred as a specialized region responsible for placental invasion and the inverted yolk sac facilitates substance exchange with the main placenta. Finally, precocial newborns evolved as a derived condition within Rodentia. All things considered, a mode of reproduction is indicated, which does not demand excessive additional energy intake by the mother and is in accordance with her low energetic diet. Hystricognathi possess major character transformations that represent prerequisites for their successful radiation at the time when more open ecosystems and grasslands evolved during Earth history. The analysis resulted in the reconstruction of a life-near picture of the hystricognath stem species pattern with high explanatory power in terms of changes in space and time and their interdependence with biodiversity.

Griscelli syndrome-type 2 in twin siblings: case report and update on RAB27A human mutations and gene structure

Griscelli syndrome (GS) is a rare autosomal recessive disorder caused by mutation in the MYO5A (GS1, Elejalde), RAB27A (GS2) or MLPH (GS3) genes. Typical features of all three subtypes of this disease include pigmentary dilution of the hair and skin and silvery-gray hair. Whereas the GS3 phenotype is restricted to the pigmentation dysfunction, GS1 patients also show primary neurological impairment and GS2 patients have severe immunological deficiencies that lead to recurrent infections and hemophagocytic syndrome. We report here the diagnosis of GS2 in 3-year-old twin siblings, with silvery-gray hair, immunodeficiency, hepatosplenomegaly and secondary severe neurological symptoms that culminated in multiple organ failure and death. Light microscopy examination of the hair showed large, irregular clumps of pigments characteristic of GS. A homozygous nonsense mutation, C-T transition (c.550C>T), in the coding region of the RAB27A gene, which leads to a premature stop codon and prediction of a truncated protein (R184X), was found. In patient mononuclear cells, RAB27A mRNA levels were the same as in cells from the parents, but no protein was detected. In addition to the case report, we also present an updated summary on the exon/intron organization of the human RAB27A gene, a literature review of GS2 cases, and a complete list of the human mutations currently reported in this gene. Finally, we propose a flow chart to guide the early diagnosis of the GS subtypes and Chédiak-Higashi syndrome.

INFLUENCE OF ASCORBIC ACID ON EGG QUALITY

The egg quality may be influenced by some nutrients added to laying hens diets and among these nutrients, the ascorbic acid is included. According to some researches, this acid, besides of improving the quality, it can increase the egg production and weight. In addition, it reduces the quantity of cracked eggs and food intake. In order to contribute with more information, this experiment was developed. Concentrations of 100; 150 and 200mg/kg of ascorbic acid were added to rations of laying hens during 60 days, with the objective of determining the influence of ascorbic acid on the quality of eggs kept under room temperature conditions (26.7 ± 2.2°C and 60 ± 5% moisture) for a period of 28 days. A total of 1440 eggs were used, being 720 of white shell and 720 of brown shell. Egg quality was determined by Haugh unit and yolk index, every 7 days. The obtained results allowed to conclude that ascorbic acid, independent on the level of supplementation, was not enough to decrease.

Origanum vulgare L. essential oil as inhibitor of potentially toxigenic Aspergilli

Origanum vulgare L. essential oil has been known as an interesting source of antimicrobial compounds to be applied in food conservation. In this study, the effect of O. vulgare essential on the growth of A. flavus, A. parasiticus, A. fumigatus, A. terreus and A. ochraceus was assessed. The essential oil had a significant inhibitory effect on all assayed fungi. MIC was 0.6 µL.mL-1 for all fungi, while MFC was in the range of 1.25-2.5 µL.mL-1. The radial mycelial growth of A. flavus and A. parasiticus was strongly inhibited over 14 days at 0.6, 1.25 and 2.5 µL.mL-1 of oil in solid medium. The mycelial mass of all fungi was inhibited over 90% at 0.6 and 0.3 µL.mL-1 in liquid medium, while it was 100% at 1.25 µL.mL-1. The oil in a range of concentrations (0.6 to 2.5 µL.mL-1) was effective in inhibiting the viability and spores germination in a short time of exposure. The main morphological changes caused by the essential oil in A. parasiticus observed under light microscopy were absence of conidiation, leakage of cytoplasm, loss of pigmentation, and disrupted cell structure. These results demonstrated that O. vulgare essential oil produced a significant fungitoxic effect supporting its possible rational use as anti-mould compound in food conservation.

Elimination of Salmonella enterica serovar Typhimurium in artificially contaminated eggs through correct cooking and frying procedures

Salmonellosis is a serious foodborne disease associated with the presence of bacteria in eggs or foods containing raw eggs. However, the use of appropriate procedures of cooking and frying can eliminate this contamination. There are few studies on the elimination of contamination of Salmonella in hens' eggs through typical frying procedures, especially for Salmonella enterica serovar Typhimurium (or S. typhimurium). The aim of this study was to determine the appropriate conditions for cooking and frying hens' eggs artificially contaminated with S. typhimurium, making them free of bacterial contamination. Hens' eggs were artificially contaminated with S. typhimurium and subjected to various processes of cooking, frying and food preparation. It was observed that the minimum time necessary to eliminate contamination through cooking procedures is 5 minutes after the water starts boiling, and also that, cooking in the microwave oven complete eliminates the bacterial contamination. When the eggs were fried on both sides, keeping the yolk hard, a complete bacterial elimination was observed. Mayonnaise prepared with vinegar presented a decrease in bacterial colonies when compared mayonese prepared with lemon.

Simultaneous α-amylase and protease production by the soil bacterium Bacillus sp. SMIA-2 under submerged culture using whey protein concentrate and corn steep liquor: compatibility of enzymes with commercial detergents

Protease and α-amylase production by a thermophilic Bacillus sp. SMIA-2 cultivated in liquid cultures containing 0.25% (w/v) starch as a carbon source reached a maximum at 18 hours (47 U.mg-1 Protein) and 36 hours (325 U.mg-1 Protein), respectively. Culture medium supplementation with whey protein concentrate (0.1%, w/v) and corn steep liquor (0.3%, w/v) not only improved the production of both enzymes but also enabled them to be produced simultaneously. Under these conditions, α-amylase and protease production reached a maximum in 18 hours with levels of 401 U.mg-1 protein and 78 U.mg-1 protein, respectively. The compatibility of the enzymes produced with commercial laundry detergent was investigated. In the presence of Campeiro® detergent, α-amylase activity increased while protease activity decreased by about 27%. These enzymes improved the cleaning power of Campeiro® detergent since they were able to remove egg yolk and tomato sauce stains when used in this detergent.

Type of dietary lipids and storing time on egg stability

The objective of this work was to evaluate the influence of diets containing different lipid sources on eggs quality during refrigerated storage, on yolk fatty acid composition, and on cholesterol in the yolk. Four diets were used containing Soy Oil (SO), Sunflower Seed (SS), and Meat and Bone Meal + Soy Oil (MBM + SO) or Meat and Bone Meal + Tallow (MBM + TA). The experiment followed a factorial design 4 × 3 with four dietary treatments and three storage times. The eggs were stored at 4 °C for 0, 30, and 60 days. The collected eggs were analyzed for egg weight loss, Haugh units, yolk moisture, yolk lipid oxidation, and cooked yolk firmness. Refrigerated storage reduced Haugh units, and increased yolk moisture. Sixty days of storage time reduced the firmness of hard-cooked yolk. There was an interaction between dietary treatment and storage time for egg weight loss and lipid oxidation. With regard to yolk fatty acid profile, MBM + TA diet increased the contents of palmitic and palmitoleic acids. The levels of oleic and arachidonic acids were higher in yolks from birds fed with SS diet. Linoleic acid level was higher in the yolk from treatment with SO diet. Diets containing MBM + SO induced higher levels of docosahexaenoic acid. Yolk cholesterol content was reduced with the inclusion of SS in the diet. Therefore, the type of lipid present in the diet and refrigerated storage for 60 days at 4 °C can affect the egg quality.

Viscosity of egg white from hens of different strains fed with commercial and natural additives

Yolk color and egg white (albumen) cleanliness and viscosity are important parameters by which consumers judge the quality of eggs. This study aimed to investigate changes in albumen viscosity during storage of eggs for up to 36 days from two different commercial laying hen strains (Carijo Barbada and Isa Brown) fed a diet containing annatto (1.5 and 2.0%) or a synthetic additive without synthetic colorants (control). Analyses of humidity, albumen height, pH, viscosity, foam formation, and stability were carried out on eggs. Carijo Barbada strain had smaller albumen, lower humidity and higher egg white viscosity than Isa Brown strain; however, with storage, viscosity lowered significantly on both strains. Initially, the addition of 2.0% of annatto or a synthetic additive increased viscosity in both strains, but with storage only the control maintained longer viscosity. Lower viscosity did not change foam density and stability.

Comparison between freeze and spray drying to obtain powder Rubrivivax gelatinosus biomass

The use of colorants in products of animal origin is justified by the improvement in the color of foods since this attribute is considered a quality criterion. These additives can be produced using industrial effluents as substrates and appropriate organisms, such as Rubrivivax gelatinosus. Oxycarotenoids represent a class of carotenes responsible for the pigmentation of animals and vegetables. R. gelatinosus grows in fish industry effluent with the resulting production of a bacterial biomass containing oxycarotenoids. The purpose of this study was to compare the use of two drying processes - spray and freeze drying - to obtain powder biomass in terms of the process parameters (yield, productivity, and product recovery) and the product characteristics (color, proximate composition, and oxycarotenoids). No difference was detected in the yield between these techniques, while productivity was higher using spray drying. Higher product recovery and moisture were achieved with freeze drying, while ash was higher with spray drying. The freeze dried biomass was redder, darker and less saturated than the spray dried biomass. No difference in oxycarotenoids was detected between the biomasses. Although it results in lower recovery rate, spray drying was faster and more productive, and it provided the same yield as freeze drying, which makes it the method of choice for obtaining R. gelatinosus biomass.

Sensory and physico-chemical characteristics of desserts prepared with egg products processed by freeze and spray drying

In this work, three freeze-dried (FD) egg products (whole egg (WE), egg yolk (EY) and egg white (EW)) were obtained and the acceptability of confections prepared with each was evaluated. Sensory analyses for confections were performed by hedonic testing with fifty panelists in each evaluation. The studied confections were: Condensed Milk Pudding (P), Quindim (Q) and Meringue (M). The results obtained for confections made with FD egg products were compared with the achieved through other formulations of the same desserts made with fresh (F) or spray-dried (SD) egg products. The sensory analysis results for confections made with FD egg products showed good acceptance by panelists. A principal component analysis of the sensory evaluation data was carried out to identify similarities between the different egg products. The PCA supported the conclusion that FD egg products can substitute their fresh and SD counterparts in dessert formulations with good acceptability while keeping the advantages conferred by the freeze-drying method.

Particle size and cholesterol content of a mayonnaise formulated by OSA-modified potato starch

Egg yolk was partially replaced (0, 25, 50, 75, and 100%) with octenyl succinic anhydride (OSA)-modified potato starch in a reduced-fat mayonnaise formulation to curtail the problems associated with high cholesterol and induced allergic reactions. The physicochemical properties included parameters such as: pH, fat content, and emulsion stability of the formulations analyzed. The samples with 75% and 100% egg yolk substitute showed the maximum emulsion stability (>95% after two of months storage), and they were selected according to cholesterol content, particle size distributions, dynamic rheological properties, microstructure, and sensory characteristic. A significant reduction (84-97%) in the cholesterol content was observed in the selected samples. Particle size analysis showed that by increasing the amount of OSA starch, the oil droplets with the peak size of 70 µm engulfed by this compound became larger. The rheological tests elucidated that in the absence of egg yolk, OSA starch may not result in a final product with consistent texture and that the best ratio of the two emulsifiers (OSA starch/egg yolk) to produce stable reduced-fat, low cholesterol mayonnaise is 75/25. The microscopic images confirmed the formation of a stable cohesive layer of starch surrounding the oil droplets emulsified in the samples selected.