Estudo da produção de biossurfactantes sintetizados por Pseudomonas aeruginosa AP029-GVIIA utilizando manipueira como fonte de carbono

This study evaluates the biosurfactants production from cassava wastewater, an agro industrial residue, to be used as carbon source. Using a factorial design 24-1 (half fraction), 10 tests were performed using Pseudomonas aeruginosa AP029/GVII-A in submerged batch cultivation in rotating incubator (shaker). The influence of factors (temperature, agitation, aeration ratio and concentration of cultivation medium) at two different levels for the synthesis of the biosurfactant. Samples were collected throughout the cultivation by 132 hours of fermentation were completed. The best outcome was intended by following production through substrate consumption, dry matter, reduction of surface tension (ring method) and emulsification index. The kinetics of microorganism was assessed for the carbon source used. The results showed that the cassava wastewater is a well assimilable substrate for the production of biotensoactive, reaching 91 % of consumption by the micro-organism under study. The growth temperature was found to be one of the leading factors in the synthesis of the metabolite, followed by aeration and also due to the agitation. The best results showed a 30 % reduction in surface tension (% RTS) for the environment, reaching values of 30 mN/m; 3.0 g /L of biomass and emulsifying index greater than 65 %. The metabolite synthesized still remained stable for different salt concentrations (1, 5 and 10 % w/ v) and alkaline pH (8-10).

Otimização do meio de cultura para a produção de quitosanase por metarhizium anisopliae em cultivo descontínuo submerso

In this work a Plackett-Burman Design with 8 factors and 12 trials in 2 levels with 3 repetitions at the center point was used in order to investigate the influence of the concentration of chitosan, peptone, yeast extract, NaNO3, K2HPO4, KCl, MgSO4.7H2O and FeSO4 on chitosanase production by Metarhizium anisopliae. Runs were carried out using submerged discontinuous cultivation for enzyme production. The results of the Plackett & Burman Design showed that only two factors, chitosan concentration as well as FeSO4 had influence on chitosanolytic activity, while the increase in concentration of other factors not contributed significantly to the quitosanolítica activity. Cultivation medium optimization for enzyme production was carried out using a Composite Central Design, with the most important factors for chitosanolytic activity (chitosan and FeSO4), in accordance with Plackett & Burman Design, and keeping the other nutrients in their minimum values. On this other design, it was taken the highest limit in Plackett & Burman Design as the lowest limit (-1) to FeSO4 factor. The results showed that the enzyme production was favoured by increasing the chitosan concentration and by decreasing FeSO4. Maximum production for chitosanolytic activity was about 70.0 U/L and was reached in only 18 h of fermentation, a result about twenty-eight times greater than a former study using the same microorganism (about 2.5 U/L at 48 h)

Production and action pattern of inulinase from Aspergillus Niger-245: hydrolysis of inulin from several sources

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

Effects of probiotic bacteria, isoflavones and simvastatin on lipid profile and atherosclerosis in cholesterol-fed rabbits: a randomized double-blind study

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

Effects of different disinfection treatments on the natural microbiota of lettuce

In this study, water and eight sanitizing solutions (vinegar at 6, 25, and 50%; acetic acid at 2 and 4%; peracetic acid at 80 ppm, sodium hypochlorite at 200 ppm, and sodium dichloroisocyanurate at 200 ppm) were compared in terms of their effectiveness against the natural microbiota of lettuce. All of the samples were kept in contact with the sanitizing solutions for 15 min, and the effectiveness of a sanitizing agent was evaluated on the basis of the number of decimal reductions of the total aerobic mesophilic count, the mold and yeast count, the total coliform count, and the Escherichia coli count. The average initial levels of these organisms in the samples were 6.94 log(10) CFU/g for aerobic mesophilic microorganisms, 5.62 log(10) CFU/g for molds and yeasts, and 3.25 log(10) CFU/g for total coliforms. of 10 samples analyzed, only 4 contained E. coli, and the average initial level of this microorganism in these 4 samples was 1.64 log(10) CFU/g. Salmonella was not detected in any of the samples tested. The decimal reductions of the populations of aerobic mesophilic microorganisms, molds and yeasts, total coliforms, and E. coli were 0.78, 0.87, 0.82, and >0.14 log(10) CFU/g, respectively, in water; 2.89, >3.41, >2.21, and >0.26 log(10) CFU/g, respectively, in 50% vinegar; 2.42, >3.20, >1.99, and >0.26 log(10) CFU/g, respectively, in 25% vinegar; 1.83, 2.57, 1.58, and >0.26 log(10) CFU/g, respectively, in 6% vinegar; 3.91, >3.58, >2.25, and >0.26 log(10) CFU/g, respectively, in 4% acetic acid; 3.37, >3.53, >2.25, and >0.26 log(10) CFU/g, respectively, in 2% acetic acid; 1.85, 2,32, 1.44, and >0.20 log(10) CFU/g, respectively, in 80 ppm of peracetic acid; 2.63, 2.75, 1.91, and >0.26 log(10) CFU/g, respectively, in 200 ppm of sodium hypochlorite; and 3.23, >3.08, >1.95, and >0.26 log(10) CFU/g, respectively, in 200 ppm of sodium dichloroisocyanurate. Statistical analysis of the results showed that the effectiveness levels for all of the sanitizing agents tested were equivalent to or higher than that for sodium hypochlorite at 200 ppm.

Cyclodextrin glycosyltransferase from Bacillus licheniformis: Optimization of production and its properties

Cyclodextrin glycosyltransferase (EC 2.4.1.19) is an enzyme that produces cyclodextrins from starch via an intramolecular transglycosylation reaction. An alkalophilic Bacillus strain, isolated from cassava peels, was identified as Bacillus licheniformis. CGTase production by this strain was better when potato starch was used as carbon source, followed by cassava starch and amylopectin. Glucose and amylose, on the other hand, acted as synthesis repressors. When the cultivation was supplemented with sodium ions and had the pH adjusted between 6.0 and 9.0, the microorganism maintained the growth and enzyme production capacity. This data is interesting because it contradicts the concept that alkalophilic microorganisms do not grow in this pH range. After ultrafiltration-centrifugation, one protein of 85.2 kDa with CGTase activity was isolated. This protein was identified in plates with starch and phenolphthalein. Determination of the optimum temperature showed higher activities at 25 degrees C and 55 degrees C, indicating the possible presence of more than one CGTase in the culture filtrate. Km and Vmax values were 1.77 mg/mL and 0.0263 U/mg protein, respectively, using potato starch as substrate.

Development and validation of a stability-indicative agar diffusion assay to determine the potency of linezolid in tablets in the presence of photodegradation products

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

Chemical and microbiological evaluation of the internal surfaces of aluminum tubes both unlined and lined with epoxi resin by means of the stereoscope and scanning electron microscope

A superfície interna das bisnagas fabricadas com alumínio não revestido e revestido com resina epóxi, utilizadas para acondicionar cremes, pomadas, géis, etc., foram avaliadas quimicamente e por métodos microbiológicos correlacionados com a aderência de microrganismos. A prova da porosidade e da resistência à remoção da resina foi observada por meio do microscópio eletrônico de varredura (Topcon FM300) e estereoscópio Leica (MZ12) acoplado a Sistema de Digitalização de Imagens. Para avaliar a ação dos microrganismos foram utilizados corpos-de-prova esterilizados (discos de 10mm de diâmetro), imersos em caldo Mueller Hinton (Difco) e colocados em tubos de polipropileno com tampa de rosca (Corning). Foram inoculados tubos com meio de cultura para cada uma das suspensões bacterianas (10(9)UFC/mL) de Streptococcus agalactiae, Staphylococcus aureus, Acinetobacter lwoffii e Candida albicans, incubados a 37°C, sob agitação constante durante 12 dias. O meio de cultura era trocado a cada 3 dias. Após esse período, os corpos-de prova foram removidos, processados e observados em microscópio eletrônico de varredura JEOL-JSM (T330A). A observação por meio do microscopio eletrônico de varredura mostrou a aderência e a formação de biofilme sobre a superfície de alumínio não revestido e revestido com resina epóxi.

Screening and Production Study of Microbial Xylanase Producers from Brazilian Cerrado

Hemicelluloses are polysaccharides of low molecular weight containing 100 to 200 glycosidic residues. In plants, the xylans or the hemicelluloses are situated between the lignin and the collection of cellulose fibers underneath. The xylan is the most common hemicellulosic polysaccharide in cell walls of land plants, comprising a backbone of xylose residues linked by beta-1,4-glycosidic bonds. So, xylanolytic enzymes from microorganism have attracted a great deal of attention in the last decade, particularly because of their biotechnological characteristics in various industrial processes, related to food, feed, ethanol, pulp, and paper industries. A microbial screening of xylanase producer was carried out in Brazilian Cerrado area in Selviria city, Mato Grosso do Sul State, Brazil. About 50 bacterial strains and 15 fungal strains were isolated from soil sample at 35 A degrees C. Between these isolated microorganisms, a bacterium Lysinibacillus sp. and a fungus Neosartorya spinosa as good xylanase producers were identified. Based on identification processes, Lysinibacillus sp. is a new species and the xylanase production by this bacterial genus was not reported yet. Similarly, it has not reported about xylanase production from N. spinosa. The bacterial strain P5B1 identified as Lysinibacillus sp. was cultivated on submerged fermentation using as substrate xylan, wheat bran, corn straw, corncob, and sugar cane bagasse. Corn straw and wheat bran show a good xylanase activity after 72 h of fermentation. A fungus identified as N. spinosa (strain P2D16) was cultivated on solid-state fermentation using as substrate source wheat bran, wheat bran plus sawdust, corn straw, corncob, cassava bran, and sugar cane bagasse. Wheat bran and corncobs show the better xylanase production after 72 h of fermentation. Both crude xylanases were characterized and a bacterial xylanase shows optimum pH for enzyme activity at 6.0, whereas a fungal xylanase has optimum pH at 5.0-5.5. They were stable in the pH range 5.0-10.0 and 5.5-8.5 for bacterial and fungal xylanase, respectively. The optimum temperatures were 55C and 60 A degrees C for bacterial and fungal xylanase, respectively, and they were thermally stable up to 50 A degrees C.

Effect of indomethacin on the pregnant rat

O objetivo deste trabalho foi avaliar a performance reprodutiva, estudo morfológico do fígado e característicapost mortem de ratas Wistar prenhes tratadas com indometacina, um inibidor geral de COX. Indometacina foi administrada oralmente, nas doses de 0 (controle), 0,32, 1,68 e 8,40 mg/kg/dia (n=10/grupo), nos dias 3 e 4 de prenhez (dia 0 = primeiro dia de prenhez = esperma positivo). Os animais foram eutanasiados sob anestesia no 11º dia de prenhez, e foram realizadas necropsia e cultura de microorganismos. Os resultados mostraram que as doses de 0,32 e 1,68 mg/kg de peso corpóreo (dose terapêutica para humanos) de indometacina não causaram efeitos embriotóxicos ou letais. A maior dose (8,40 mg/kg) de indometacina prejudicou o processo de implantação e, portanto, interferiu no desenvolvimento fetal. A peritonite foi detectada na necropsia e nos estudos bacteriológicos dos animais tratados com 8,4 mg/kg e considerada a causa-morte destes animais. Portanto, este estudo analisou um agente farmacológico na prenhez de roedores e evidenciou que a indometacina apresentou efeitos embriotóxicos e letais na maior dose empregada, mas foi segura na dose terapêutica usada pelo homem.

Lack of Epstein-Barr virus infection in cervical carcinomas

Context.-The Epstein-Barr virus (EBV) is a ubiquitous microorganism strongly associated with lymphoproliferative disorders and a large number of human neoplasms, mainly undifferentiated nasopharyngeal carcinoma and Burkitt lymphoma. The viral DNA has been detected in other tumors, such as carcinomas from tonsil, salivary glands, and thymus, and malignancies of the female genital tract. Some authors have proposed that EBV could play a role in the carcinogenesis of cervical tumors; however, other studies do not support this hypothesis.Objective.-To assess whether EBV is associated with female genital tract neoplasms.Design.-Sixty-five biopsy specimens (5 in situ carcinomas, 24 invasive squamous cell carcinomas, 6 lymphoepithelioma-like carcinomas, and 30 endocervical adenocarcinomas) were used to perform EBV detection through RNA in situ hybridization.Results.-None of the cervical carcinoma cases studied was positive for EBV infection.Conclusions.-The results suggest that it is still premature to incriminate EBV in the carcinogenesis of cervical carcinoma.

Comparison between qualitative and semiquantitative catheter-tip cultures: laboratory diagnosis of catheter-related infection in newborns

Este estudo prospectivo avaliou os métodos semiquantitativo e qualitativo de cultura de cateter para o diagnóstico de infecção relacionada a cateter (IRC) em recém-nascidos (RN). Foram incluídas pontas de cateteres provenientes de recém-nascidos internados na Unidade Neonatal do Hospital das Clínicas da Faculdade de Medicina de Botucatu, UNESP. Foram utilizadas as técnicas semiquantitativa e qualitativa de cultura de cateter. Para o diagnóstico de IRC, os microrganismos isolados das culturas de cateteres e de hemoculturas periféricas foram identificados e submetidos ao teste de sensibilidade a antimicrobianos. O padrão ouro correspondeu ao diagnóstico de certeza de IRC, com o isolamento do mesmo microrganismo (espécie e perfil de sensibilidade a antimicrobianos) isolado em hemocultura periférica. Foram estudados 85 cateteres provenientes de 63 RN. A cultura semiquantitativa, embora tenha apresentado menor sensibilidade (90%), apresentou uma maior especificidade (71%) em comparação à sensibilidade de 100% e especificidade de 60% encontradas na cultura qualitativa. Através da identificação dos microrganismos obtidos nas culturas de cateteres, observou-se uma predominância de espécies de Estafilococos coagulase-negativa (ECN). A espécie Staphylococcus epidermidis foi a prevalente (77,5%) nos cateteres com culturas semiquantitativas positivas. Dos 11 episódios de IRC diagnosticados, 8 (72,7%) foram associados a espécies de ECN, dos quais 6 eram da espécie S. epidermidis. Também foram detectados dois casos de IRC por S. aureus e um caso por Candida parapsilosis. O método de cultura semiquantitativo cateter apresentou vantagens para o diagnóstico de IRC em RN quando comparado com o método qualitativo tradicional.

Correlation of Atopobium vaginae Amount With Bacterial Vaginosis Markers

Objective. The purpose of this study was to test the correlation of the amount of Atopobium vaginae with the most commonly used markers for bacterial vaginosis (BV).Materials and Methods. We enrolled 103 nonpregnant and premenopausal women that were positive for BV by Amsel criteria and with a Nugent score higher than 3. All women were negative for yeast, Chlamydia trachomatis, Trichomonas vaginalis, and Neisseria gonorrhoeae. A. vaginae concentration was determined by quantitative polymerase chain reaction from samples of vaginal rinsings with 2 mL of sterile saline.Results. There was no difference in the median values of A. vaginae concentration when comparing samples with presence or absence of each individual Amsel criterion. In the case of a higher pH cutoff value of 4.9, greater amounts of this microorganism (p = .02) were found. In addition, correlation tests showed that A. vaginae concentration is positively correlated with pH (p < .001) and with Nugent scores (p = .003).Conclusions. The quantification of A. vaginae is useful for identification of the most severe cases of BV.

Correlation between API 50 CH and multiplex polymerase chain reaction for the identification of vaginal lactobacilli in isolates

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

Cloning, sequencing and expression analysis of the equine hepcidin gene by real-time PCR

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