Imunolocalização do fator de crescimento semelhante à insulina (IGF-I) e do fator de crescimento epidermal (EGF) em ovários caninos e seus efeitos sobre a maturação in vitro de oócitos: avaliação nuclear

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Efficacy of sperm motility after processing and incubation to predict pregnancy after intrauterine insemination in normospermic individuals

Background: Intrauterine insemination (IUI) is widely used to treat infertility, and its adequate indication is important to obtain good pregnancy rates. To assess which couples could benefit from IUI, this study aimed to evaluate whether sperm motility using a discontinuous gradient of different densities and incubation in CO2 in normospermic individuals is able to predict pregnancy.Methods: A total of 175 couples underwent 175 IUI cycles. The inclusion criteria for women were as follows: 35 years old or younger (age range: from 27 to 35 years) with normal fallopian tubes; endometriosis grades I-II; unexplained infertility; nonhyperandrogenic ovulatory dysfunction. Men with normal seminal parameters were also included. All patients underwent ovarian stimulation with clomiphene citrate and human hMG or r-FSH. When one or (at most) three follicles measuring 18 to 20 mm were observed, hCG (5000 UI) or r-hCG (250 mcg) was administered and IUI performed 36-40 h after hCG. Sperm processing was performed using a discontinuous concentration gradient. A 20 microliters aliquot was incubated for 24 h at 37 degrees C in 5% CO2 following a total progressive motility analysis. The Mann-Whitney and Chi-square tests, as well as a ROC curve were used to determine the cutoff value for motility.Results: Of the 175 couples, 52 (in 52 IUI cycles) achieved clinical pregnancies (CP rate per cycle: 29.7%). The analysis of age, duration and causes of infertility did not indicate any statistical significance between pregnancy and no pregnancy groups, similar to the results for total sperm count and morphology analyses, excluding progressive motility (p < 0.0001). The comparison of progressive motility after processing and 24 h after incubation between these two groups indicated that progressive motility 24 h after incubation was higher in the pregnancy group. The analysis of the progressive motility of the pregnancy group after processing and 24 h after incubation has not shown any motility difference at 24 h after incubation; additionally, in couples who did not obtain pregnancy, there was a statistically significant decrease in progressive motility 24 h after incubation (p < 0.0001). The ROC curve analysis generated a cutoff value of 56.5% for progressive motility at 24 h after incubation and this cutoff value produced 96.1% sensitivity, 92.7% specificity, 84.7% positive predictive value and 98.3% negative predictive value.Conclusions: We concluded that the sperm motility of normospermic individuals 24 h after incubation at 37 degrees C in 5% CO2, with a cutoff value of 56.5%, is predictive of IUI success.

Fermentação, cinética e degradação ruminal em ovinos alimentados com fontes de carboidratos associadas ao óleo

The objective this work was to evaluate the degradation, fermentation and kinetics of passage rumen fluid en sheep fed diets containing different carbohydrate associated with the oil. Four rumen cannulated sheep were allotted in 4 x 4 latin square. The treatments consisted of diets with high neutral detergent soluble fiber and diets with high starch associated or not with 4,2% of oil. Incubation times were 3, 6, 12, 24, 48, 72, 96 and 120 hours and ruminal fluid was collected at 0, 2, 4, 6, 8, 10, 12, and 24 hours after feeding. Diets high neutral detergent fiber soluble resulted in greater degradation of dry matter, crude protein, neutral detergent fiber soluble and starch, and higher pH values, butyric acid production, dilution rate and ruminal recycling compared with diets high starch content. The inclusion of 4,2% oil the different carbohydrate sources had no influence on the fermentation kinetics and degradation of the fiber. High content of neutral detergent soluble fiber in the diet favors the ruminal fermentation compared to the high starch content.

Atividade antimicrobiana de enxaguatórios bucais sem álcool à base de clorexidina sobre candida albicans

The aim of this study was to evaluate the antimicrobial activity of alcohol-free mouthwashes on Candida albicans. Twenty clinical isolates of C. albicans and one reference strain (ATCC 18804) were evaluated after exposure to two 0.12% chlorhexidine-based and alcohol-free (“Ca” and “Or”) in comparison to gluconate chlorhexidine with ethanol (positive control). The maximum inhibitory dilution (MID) and maximum fugal dilution (MFD) were determined by the microdilution method. Twelve serial dilutions (from 50 to 0.02%) were prepared in duplicate. Then, 100 µL of C. albicans suspension (106 cells.mL–1) were added to the wells. After incubation (37 °C/24 hours), MID was determined by reading the optical density. For MFD determination, the content of the wells were plated on Saouraud agar. For MID, there were no differences between groups Or and control, but Ca group showed a MID statistically higher (Kruskal-Wallis, p = 0.0012). For MFD, there were no differences between Ca and control (Mann-Whitney test, p = 0.1631). It can be concluded that Ca group showed a fungicid activity against C. albicans similar to the control, but lower fungistatic activity when compared to the control. Group Or showed only a fungistatic action similar to control.

Profiles of steroid hormones in canine X-Linked muscular dystrophy via stable isotope dilution LC-MS/MS

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

Response of laying hens to methionine plus cystine intake by dilution technique

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

Incubation temperature manipulation during fetal development reduces adiposity of broiler hatchlings

Broilers are known as an efficient source of lean meat. Genetic selection resulted in broiler strains with large body size and fast growth, but a concomitant increase in fat deposition also occurred. Other than reducing nutrient intake, there is a lack of alternative methods to control body fat composition of broilers. The present study assessed whether incubation temperature (machine temperatures: 36ºC, 37.5ºC, and 39ºC; eggshell temperatures: 37.4 ± 0.08°C, 37.8 ± 0.15ºC, and 38.8 ± 0.33°C, respectively.) from d 13 affects broiler hatchling fat deposition. We analyzed adipocyte hypertrophy and proliferation in 3 body regions; weight and chemical composition of yolk-free chicks and yolk sacs; and serum lipid profile. Increased incubation temperature reduced abdominal and cervical adipocyte size. Independently of temperature, cervical adipocytes were smaller and showed higher proliferation than adipocytes in the abdominal and thigh regions. Smaller cervical adipocytes were observed in birds from eggs incubated at 36ºC and 39ºC. With regard to weight and composition of chicks, ash content as a percentage of dry matter was the only variable affected by temperature; it was higher in chicks from eggs incubated at 36ºC than at 39ºC and showed no significant difference between chicks incubated at 39ºC and 37.5ºC. Absolute and relative weights of yolk sacs were higher from eggs incubated at 39ºC than at 36ºC, and these two treatments did not differ from the 37.5ºC control. Absolute measures of yolk sac lipids, moisture, dry matter, and crude protein content were lower in chicks from eggs incubated at 36ºC, and no significant differences were found for these variables between chicks from eggs incubated at 37.5ºC and 39ºC. Hatchlings from eggs incubated at 36°C had significantly higher cholesterol levels than chicks incubated at the other 2 temperatures, but no additional effects on blood lipids were detected. Incubation temperature manipulation during fetal development altered cervical and abdominal adipocyte size in broiler hatchlings and could become a tool in hatcheries to manipulate chick quality, although further studies are needed to evaluate its long-term effects.

In vitro reduction of Streptococcus mutans biofilm on silver nanoparticle-modified resin composite

Introduction: Currently, new methods to reduce biofilm formation on biomaterials are very studied, for example the use of silver nanoparticles, which were bactericidal. However, there are few studies investigating the benefits of these particles in dental restorative materials. Objective: This study aimed to compare in vitro the Streptococcus mutans biofilm formation on conventional light-cured composite resin with that on experimental light-cured composite resin, modified with silver nanoparticles. Material and methods: Discs were produced with either conventional resin (control group) and resin modified with different concentrations of silver nanoparticles, 0.1%, 0.3% and 0.6 % wt. (groups 1, 2 and 3, respectively). The samples were incubated in bacterial suspension (S. mutans) enriched with 20% sucrose to promote biofilm growth on the surfaces. Incubation times were 1, 4 and 7 days. After each period, adherent biofilms were disaggregated by ultrasound. Then, the numbers of viable cells recovered from the biofilms were counted through the serial dilution method. A morphological analysis of biofilm was also performed by Scanning Electron Microscopy. The data were subjected to Anova and Tukey’s test (α = 0.05). Results: The number of viable cells was statistically lower in groups 2 and 3 than in group 1 and control group, after the three incubation periods, without statistical difference between groups 2 and 3. The number of viable cells was statistically lower in group 1 than in control group, after 4 and 7 days of incubation. Conclusion: Resins modified with silver presented reduction of S. mutans biofilm on their surfaces, according to the conditions of this study.

Quantificação do fator de crescimento semelhante a insulina I (IGF-I) em plasma bovino por ELISA

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Occurrence of Methanogenic Archaea in Highly Polluted Sediments of Tropical Santos-So Vicente Estuary (So Paulo, Brazil)

Little is known about the ability of methanogens to grow and produce methane in estuarine environments. In this study, traditional methods for cultivating strictly anaerobic microorganisms were combined with Fluorescence in situ hybridization (FISH) technique to enrich and identify methanogenic Archaea cultures occurring in highly polluted sediments of tropical Santos-So Vicente Estuary (So Paulo, Brazil). Sediment samples were enriched at 30A degrees C under strict anaerobic and halophilic conditions, using a basal medium containing 2% of sodium chloride and amended with glucose, methanol, and sodium salts of acetate, formate and lactate. High methanogenic activity was detected, as evidenced by the biogas containing 11.5 mmol of methane at 20 days of incubation time and methane yield of 0.138-mmol CH(4)/g organic matter/g volatile suspense solids. Cells of methanogenic Archaea were selected by serial dilution in medium amended separately with sodium acetate, sodium formate, or methanol. FISH analysis revealed the presence of Methanobacteriaceae and Methanosarcina sp. cells.

In vitro evaluation of the microbial contamination on new toothbrushes: A preliminary study

Objective: The presence and survival of microorganisms on toothbrush bristles might play a role on the etiology of oral infections. The aim of this in vitro study was to evaluate the presence of bacterial contamination on new toothbrushes before oral contact. Materials and methods: Forty toothbrushes from five different manufacturers were used in this experimental study. Each manufacturer was divided according to conventional local of obtaining: industry, drugstore, market, and perfumery. The toothbrush heads were completely immersed into tubes containing 5.0 mL of sterile peptonated water (dilution 1:10). A group of eight tubes containing the sterile solution was used as control. After 21 days of anaerobic incubation, occurrence of contamination was visually evaluated and confirmed by light microscopy. Results: Bacterial growth in the medium, indicative of bristles contamination, was found in a total of 19 out of 40 samples (47.5%) evaluated: 6 out of 14 samples (42.85%) from industry group, 4 out of 8 samples (50.0%) from drugstore, 5 out of 10 samples (50.0%) from market, and 4 out of 8 samples (50.0%) from perfumery. Only the toothbrushes with bristles coated with chlorhexidine did not show contamination. The Gram-negative sporulating bacilli were the most prevalent form recovered. Conclusions: Except for chlorhexidine group, bacterial growth was observed in all groups evaluated irrespective local of obtaining. Microsc. Res. Tech., 2012. (c) 2011 Wiley Periodicals, Inc.

Bioelectrical impedance with different equations versus deuterium oxide dilution method for the inference of body composition in healthy older persons

There is no consensus regarding the accuracy of bioimpedance for the determination of body composition in older persons. This study aimed to compare the assessment of lean body mass of healthy older volunteers obtained by the deuterium dilution method (reference) with those obtained by two frequently used bioelectrical impedance formulas and one formula specifically developed for a Latin-American population. A cross-sectional study. Twenty one volunteers were studied, 12 women, with mean age 72 +/- 6.7 years. Urban community, Ribeiro Preto, Brazil. Fat free mass was determined, simultaneously, by the deuterium dilution method and bioelectrical impedance; results were compared. In bioelectrical impedance, body composition was calculated by the formulas of Deuremberg, Lukaski and Bolonchuck and Valencia et al. Lean body mass of the studied volunteers, as determined by bioelectrical impedance was 37.8 +/- 9.2 kg by the application of the Lukaski e Bolonchuk formula, 37.4 +/- 9.3 kg (Deuremberg) and 43.2 +/- 8.9 kg (Valencia et. al.). The results were significantly correlated to those obtained by the deuterium dilution method (41.6 +/- 9.3 Kg), with r=0.963, 0.932 and 0.971, respectively. Lean body mass obtained by the Valencia formula was the most accurate. In this study, lean body mass of older persons obtained by the bioelectrical impedance method showed good correlation with the values obtained by the deuterium dilution method. The formula of Valencia et al., developed for a Latin-American population, showed the best accuracy.

Evaluation of Inductively Coupled Plasma Mass Spectrometry for Determining Ca, Cu, Fe, Mg, Mn, Se and Zn in Bovine Semen Samples using a Simple Sample Dilution Method

A simple and fast method for the determination of Ca, Cu, Fe, Mg, Mn, Se and Zn in bovine semen by quadrupole inductively coupled plasma spectrometry (q-ICP-MS) is described. Prior to analysis, samples (200 mu L) were diluted 1:50 in a solution containing 0.01% v/v Triton (R) X-100 and 0.5% v/v nitric acid and directly analyzed by ICP-MS. The limits of detection of the method are 0.3, 0.03, 0.2, 0.04, 0.04, 0.03 and 0.03 mu g L-1 for Ca-44, Cu-63, Fe-57, Mg-24, Zn-64, Se-82 and Mn-55, respectively. For purposes of comparison and method validation, four ordinary bovine semen samples were directly analyzed by ICP-MS and by flame atomic absorption spectrometry (FAAS) or graphite furnace atomic absorption spectrometry (GF AAS), with no statistical difference between the techniques at the 95% level when applying the t-test. Then, the proposed method was applied in the determinations of Ca, Cu, Fe, Mg, Mn, Se and Zn in collected samples of bovine semen from different breeds, which are used in reproduction programs and artificial insemination.

Evaluation of inductively coupled plasma mass spectrometry for determining Ca, Cu, Fe, Mg, Mn, Se and Zn in bovine semen samples using a simple sample dilution method

A simple and fast method for the determination of Ca, Cu, Fe, Mg, Mn, Se and Zn in bovine semen by quadrupole inductively coupled plasma spectrometry (q-ICP-MS) is described. Prior to analysis, samples (200 µL) were diluted 1:50 in a solution containing 0.01% v/v Triton® X-100 and 0.5% v/v nitric acid and directly analyzed by ICP-MS. The limits of detection of the method are 0.3, 0.03, 0.2, 0.04, 0.04, 0.03 and 0.03 µg L-1 for 44Ca, 63Cu, 57Fe, 24Mg, 64Zn, 82Se and 55Mn, respectively. For purposes of comparison and method validation, four ordinary bovine semen samples were directly analyzed by ICP-MS and by flame atomic absorption spectrometry (FAAS) or graphite furnace atomic absorption spectrometry (GF AAS), with no statistical difference between the techniques at the 95% level when applying the t-test. Then, the proposed method was applied in the determinations of Ca, Cu, Fe, Mg, Mn, Se and Zn in collected samples of bovine semen from different breeds, which are used in reproduction programs and artificial insemination.

Comparison of indigestible markers from in situ and in vivo incubation to predict apparent digestibility in hay- and corn-fed horses

Four castrated crossbred horses were used in a randomized block design to study the use of indigestible internal markers iNDF and iADF obtained in situ (from bovines) or in vivo (from equines). Treatments consisted of determining digestibility by the direct method comprising total feces collection (TC) and by the indirect method comprising internal markers iNDF and iADF obtained by in situ incubation in bovine rumen or in vivo by the mobile nylon bag (MNB) technique with horses. iNDF-IV and iADF-IV resulted in better marker recovery rate (RR) (91.50%), similar to TC. The in situ technique resulted in lower RR values for the two indigestible markers, averaging 86.50% (p < 0.05). Estimates of the nutrient coefficient of digestibility (CD) were adequately predicted by iADF-IV, for horses fed on hay exclusively, with rates 46.41, 48.16, 47.92 and 45.51% for dry matter (DM), organic matter (OM), FDN and gross energy, respectively. Results show that MNB may be used to obtain iADF in horses fed on coast-cross hay exclusively, whereas NDFi and ADFi were selected for horses fed on mixed diets to predict the coefficient of nutrient digestibility.