In vitro protein digestibility of enzymatically pre-treated bean (Phaseolus vulgaris L.) flour using commercial protease and Bacillus sp. protease

The common bean (Phaseolus vulgaris L.) is a staple food in the Brazilian diet and represents the major source of dietary protein and other micronutrients and minerals. Despite the considerable protein concentration in beans, the food is considered of low biological value when compared to animal proteins and other plant protein sources. To improve the availability of protein in beans, enzymatic treatments were performed in four cultivars (ON, OPNS, TAL and VC3). The approach was a completely randomized design with four replicates. We used a 4 × 3 factorial arrangement (four cultivars and three treatments: treatment 1-addition of commercial protease (Trypsin 250, Difco), treatment 2-addition of protease from Bacillus sp., and treatment 3:-control without enzyme addition). The enzyme: substrate ratio was 5% w/w (amount of enzyme per total protein in bean flour). The approach was a completely randomized design with four replicates. A 4 × 3 factorial arrangement (four cultivars and three treatments, the same as those mentioned above) was used. The concentration of total protein (g.100 g-1 of dry matter) in the samples ranged from 16.94 to 18.06%, while the concentration of total phenolics was between 0.78 and 1.12% (g Eq. tannic acid.100 g-1 dry matter). The in vitro protein digestibility of enzymatically untreated bean flour (control) ranged from 47.30 to 56.17% based on the digestibility of casein. Concentrations of P, K, Ca, Mg, and Zn observed in the four cultivars tested were within the average values available in the literature. Treatment 2 with protease from Bacillus sp. induced decreases in the levels of Cu and Mn. The average Fe content increased in all bean flour samples when treated with proteases, reaching a maximum increase of 102% in the TAL flour treated with protease from Bacillus sp. The digestibility of all beans tested was significantly increased (p < 0.05) after the enzyme treatment. The greatest change was observed in the OPNS cultivar treated with protease from Bacillus sp., which increased its digestibility from 54.4% (control treatment) to 81.6%.

Determination of thermobacteriological parameters and size of Bacillus stearothermophilus ATCC 7953 spores

In order to determine thermobacteriological parameters for B. stearothermophilus spores, they were diluted in a saline solution medium and in ground corn-soybean mix, distributed in TDT tube, and submitted to heat for a specific period of time. The D value (time to reduce 1 log cycle of microbial count under a certain temperature) and z value (variation of temperature to cause 10-fold change in D value) were estimated. To estimate their dimensions, the spores were visualized by using a scanning electron microscope. D121.1 ºC and z values for these spores, as determined in the saline solution, were 8.8 minutes and 12.8 ºC, respectively. D121,1 ºC and z values determined in the corn-soy mix were 14.2 minutes and 23.7 ºC, respectively. The micrographs indicated that the spores have homogeneous shape and size, with length and diameter of 2 and 1 µm, respectively. These results confirm that the spore is highly thermal-resistant, and it is a good biological indicator to evaluate the extrusion process as a feed sterilizer.

Production and partial characterization of alkaline polygalacturonase secreted by thermophilic Bacillus sp. SMIA-2 under submerged culture using pectin and corn steep liquor

Polygalacturonase production by the thermophilic Bacillus sp. SMIA-2 cultivated in liquid cultures containing 0.5% (w/v) apple pectin and supplemented with 0.3% (w/v) corn steep liquor, reached its maximum after 36 hours with levels of 39 U.mL-1. The increase in apple pectin and corn steep liquor concentrations in the medium from 0.5 and 0.3%, respectively, to 0.65%, markedly affected the production of polygalacturonase, whose activity increased four times, reaching a maximum of 150.3 U.mL-1. Studies on polygalacturonase characterization revealed that the optimum temperature of this enzyme was between 60-70 °C. Thermostability profile indicated that the enzyme retained about 82 and 63% of its activity at 60 and 70 °C, respectively, after 2 hours of incubation. The optimum pH of the enzyme was found to be 10.0. After incubation of crude enzyme solution at room temperature for 2 hours at pH 8.0, a decrease of about 29% on its original activity was observed. At pH 10.0, the decrease was 25%.

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.

Influence of xylanase addition on the characteristics of loaf bread prepared with white flour or whole grain wheat flour

The aim of this study was to verify the influence of the addition of the enzyme xylanase (four concentrations: 0, 4, 8, and 12 g.100 kg-1 flour) on the characteristics of loaf bread made with white wheat flour or whole grain wheat flour. Breads made from white flour and added with xylanase had higher specific volumes than those of the control sample (no enzyme); however, the specific volume did not differ significantly (p < 0.05) among the breads with different enzyme concentrations. All formulations made from whole grain wheat flour and added with xylanase also had specific volumes significantly higher than those of the control sample, and the highest value was found for the 8 g xylanase.100 kg-1 flour formulation. With respect to moisture content, the formulations with different enzyme concentrations showed small significant differences when compared to the control samples. In general, breads made with the addition of 8 g enzyme.100 kg-1 flour had the lowest firmness values, thus presenting the best technological characteristics.

Sanidade de sementes de arroz, biocontrole, caracterização e transmissão de Curvularia lunata em semente-plântula de arroz

Este trabalho objetivou avaliar a qualidade sanitária de sementes de arroz, quantificar a transmissão de Curvularia lunata associada à semente e à plântula, e avaliar o efeito dos isolados de Bacillus spp., no controle da mancha foliar, do cultivar Bonança. Para avaliação da sanidade de sementes, as amostras foram desinfestadas e plaqueadas, sendo avaliadas após sete dias de incubação. Na confirmação da patogenicidade, as plântulas foram inoculadas por meio de pulverização e a avaliação da doença ocorreu sete dias após. Na caracterização do isolado de C. lunata, foi realizada medição da largura, comprimento e número de septos de 100 conídios. Para a taxa de transmissão de C. lunata, foram preparadas 12 bandejas, com 100 sementes cada, de modo a proceder-se às avaliações aos 7, 14, 21 dias após a semeadura (d.a.s.). De cada uma das 100 plântulas, foram separadas as raízes, após sete dias. A inoculação dos isolados de Bacillus foi realizada na forma de suspensão, pulverizada sobre a parte aérea das plântulas. Após uma semana, foi inoculado a C. lunata. A avaliação foi efetuada aos 15 dias após inoculação dos isolados patogênicos, medindo-se o tamanho da lesão. No teste de sanidade foi detectada maior incidência de C. lunata (70%), o qual se apresentou patogênico à variedade de arroz Bonança, sendo detectado em todos os órgãos da plântula. Dentre os isolados de Bacillus, os que mais se destacaram no controle foram B41 (B. Cereus) e B35 (Bacillus sp.), confirmando seu potencial biocontrolador.

Controle da ferrugem e da broca-dos-ramos da figueira com diferentes fungicidas e inseticidas

Temperaturas e precipitações elevadas favorecem a incidência da ferrugem (Cerotelium fici(Cast.)) e da broca-dos-ramos (Azochis gripusalis (Walker, 1859) (Lepidoptera: Pyralidae)), limitando a produção comercial de figos. O objetivo deste trabalho foi comparar a eficiência de fungicidas e inseticidas alternativos em relação à de produtos convencionais registrados para a cultura. Foram realizados dois experimentos, no delineamento de blocos inteiramente casualizados, com quatro repetições, no setor de fruticultura da Universidade Tecnológica Federal do Paraná, Campus Dois Vizinhos. Para controle da ferrugem, foram utilizados, em 100 L de água, azoxistrobin (10 g), calda bordalesa (1.500 g de cal virgem + 1.500 g de sulfato de cobre) e testemunha (água). No controle da broca-dos-ramos utilizaram-se, em 100 L de água, azadiractina (1.000 mL P.C.), alho (Allium sativum L.) (100 mL P.C.), cinza (20.000 g), extrato de fumo (nicotina) (10.000 mL do preparado), deltametrina (50 mL P.C.), Bacillus thuringiensis Berliner (100 g P.C.), rotenona (1.000 mL P.C.), sabão de coco (1.000 g) e testemunha (água). A calda bordalesa foi o tratamento mais efetivo no controle da ferrugem, promovendo aumento da produtividade e da qualidade dos frutos. A deltametrina promoveu o melhor controle da broca-dos-ramos da figueira. Entre os produtos alternativos testados, o alho foi o mais efetivo no controle dessa praga.

Pesquisa de alguns microrganismos em saladas com maionese adquiridas em restaurantes, lanchonetes e "rotisseries"

Em vinte amostras de saladas com maionese foram efetuadas as contagens de bactérias mesófilas e psicrófilas, Staphylococcus aureus, Bacillus cereus, bolores e leveduras, a determinação do Número Mais Provável (NMP) de bactérias coliforme totais de Escherichia coli e de estreptococos fecais, bem como a pesquisa de salmonelas. A contagem de bactérias mesófilas variou de 2,64 x 10(4) a ³3 x 10(7)/g do produto. Quanto às bactérias psicrófilas, as contagens variaram de < 10 a ³ 3 x 10(7)/g. Para S. aureus, as contagens oscilaram de < 10² a 4 x 10(5)/g do alimento, enquanto que para B. cereus os números mínimo e máximo foram < 10² e ³ 3 x 10(4)/g, respectivamente. Para bolores e leveduras, as contagens variaram de 7,1 x 10² a 3,7 x 10(6)/g. Com relação ao NMP de coliformes totais e estreptococos fecais, os resultados obtidos mostraram-se compreendidos entre < 0,03 e ³ 4,3 x 10(5)/g. Quanto ao NMP de E. coli os números mínimo e máximo obtidos foram respectivamente de < 0,03 e ³ 2,4 x 10(4)/g de salada com maionese. Tais constatações indicam a ocorrência de contaminação inclusive por microrganismos de origem fecal. Todas as amostras revelaram-se negativas para bactérias do gênero Salmonella.

Esterilização por óxido de etileno: II. Influência de corpos de prova no desempenho de monitores biológicos e sua avaliação

Pela importância da garantia de esterilidade em produto médico-hospitalares e pela predominância da forma tubular dentre estes produtos, conduziu-se o estudo de monitores biológicos em que os corpos de prova apresentavam diferentes comprimentos e diâmetros. O sensor biológico foi constituído de esporos de Bacillus subtilis var. niger adquirido do mercado nacional sob tiras de papel. Após esterilização por processo industrial, seguiu-se a recuperação dos esporos sobreviventes através de inoculação dos suportes em caldo tioglicolato, caldo caseína-soja e este adicionado de azul de bromotimol. A capacidade promotora de crescimento destes 3 meios de cultura não apresentou diferenças. A eficácia esterilizante foi dependente do tamanho de corpos de prova. A determinação periódica do teor residual de gás nos corpos de prova, bem como a monitoração ambiental industrial indicaram a importância da legislação em vigor, devendo ser obedecida a fim de garantir inocuidade aos pacientes e operadores.

Chromobacterium violaceum infection in Brazil. A case report

We report the second case of infection with Chromobacterium violaceum that occurred in Brazil. A farm worker living in the State of São Paulo presented fever and severe abdominal pain for four days. At hospitalization the patient was in a toxemic state and had a distended and painful abdomen. Chest X-ray and abdominal ultrasound revealed bilateral pneumonia and hypoechoic areas in the liver. The patient developed failure of multiple organs and died a few hours later. Blood culture led to isolation of C. violaceum resistant to ampicillin and cephalosporins and sensitive to chloramphenicol, tetracyclin, aminoglicosydes, and ciprofloxacin. Autopsy revealed pulmonary microabscesses and multiple abscesses in the liver. The major features of this case are generally observed in infections by C. violaceum: rapid clinical course, multiple visceral abscesses, and high mortality. Because of the antimicrobial resistance profile of this Gram-negative bacillus, for appropriate empirical antibiotic therapy it is important to consider chromobacteriosis in the differential diagnosis of severe community infections in Brazil.

Pulmonary tuberculosis: relationship between sputum bacilloscopy and radiological lesions

OBJECTIVES: To determine the frequency of radiological manifestations of chest tuberculosis among the tuberculosis outpatients at the Santa Casa de Misericórdia de São Paulo Hospital, and to correlate these radiological findings with the sputum bacilloscopy. SAMPLE AND METHODS: A review was made of the medical record cards and chest X-rays of all patients attended between January 1996 and December 1998. Patients with a diagnosis of tuberculosis who presented intrathoracic manifestations of the disease and negative anti-HIV serology were selected. RESULTS: The selection included 153 patients, with an average age of 37.5 years, who were predominantly male (60.8%) and white (56.9%). Pulmonary lesions were present in 121 (79.9%) and extrapulmonary lesions in 32 (20.1%). Parenchymal-infiltrate lesions appeared in 56 patients (36.6%), cavity lesions in 55 (36.0%), pleural effusion in 28 (18.3%), isolated nodules in 6 (3.9%), mediastinal enlargement in 4 (2.6%) and miliary pattern in 4 (2.6%). Cavities were present in 45.5% of the patients with pulmonary lesions, generally in association with the parenchymal-infiltrate lesions. Parenchymal infiltrate was present in 86.8% of the patients with pulmonary lesions. There was significant presence of alcohol-acid resistant bacillus in the sputum of patients with cavities (76.4%), in comparison with those without cavities (50%) (p = 0.003). CONCLUSIONS: Parenchymal-infiltrate lesions are the most frequent radiological manifestation of pulmonary tuberculosis, and they are generally associated with cavities. There is a relationship between the presence of acid fast bacilli in sputum and pulmonary cavity lesions.

Burkholderia pseudomallei: a case report of a human infection in Ceará, Brazil

Burkholderia pseudomallei has rarely been isolated from environmental and clinical specimens in South America, particularly, in Brazil. This report describes a case of melioidosis with fulminant sepsis in a 10 year old boy, from rural area, in Tejuçuoca, State of Ceará, Brazil. Blood samples were positive and, through the analysis of results from biochemical tests and of drugs susceptibility profile, identified this gram-negative bacillus as B. pseudomallei. The contamination source remains obscure in this case, as soil and water tanks samples submitted to microbiological analyses did not indicate the presence of B. pseudomallei.

Persistence of Vectobac WDG and Metoprag S-2G against Aedes aegypti larvae using a semi-field bioassay in Rio de Janeiro, Brazil

Persistence of Bacillus thuringiensis var. israelensis (Vectobac WDG) and methoprene (Metoprag S-2G) was evaluated against Aedes aegypti late third instar larvae of the Rockefeller strain in a semi-field bioassay. Tests were performed in Rio de Janeiro, using containers made of plastic, iron, concrete and asbestos, placed in a shaded area. The formulations used were 0.2 g of Vectobac-WDG and 1g of Metoprag S-2G per 100 liters of water in house storage containers. Vectobac WDG was tested twice, in March and in April/May, 2002. In March (temperature ranging from 21.5 to 39.3 ºC), 70-100% mortality was observed by the 7th day and declined abruptly thereafter. No significant differences were observed among the container types. In April/May (18.6 to 34.8 ºC) mortality was higher than 70% to 30-36 days in all cases, except in the iron container (40% mortality on the 12th day). Metoprag S-2G was evaluated in April/May, 2002, and induced mortality higher than 70% up to 15 days in the plastic and iron containers and only seven days in the concrete container. In the asbestos container, maximal mortality was achieved on day one post-treatment (66%). Our results point to a low persistence of both formulations in the weather conditions of Rio de Janeiro.

Cutaneous anthrax in Lima, Peru: retrospective analysis of 71 cases, including four with a meningoencephalic complication

Anthrax is a zoonosis produced by Bacillus anthracis, and as an human infection is endemic in several areas in the world, including Peru. More than 95% of the reported naturally acquired infections are cutaneous, and approximately 5% of them can progress to meningoencephalitis. In this study we review the clinical and epidemiological characteristics of the patients with diagnosis of cutaneous anthrax evaluated between 1969 and 2002 at the Hospital Nacional Cayetano Heredia (HNCH) and the Instituto de Medicina Tropical Alexander von Humboldt in Lima, Peru. Seventy one patients were included [49/71 (69%) of them men], with a mean age of 37 years. The diagnoses were classified as definitive (44%) or probable (56%). The most common occupation of the patients was agriculture (39%). The source of infection was found in 63 (88.7%) patients. All the patients had ulcerative lesions, with a central necrosis. Most of the patients (65%) had several lesions, mainly located in the upper limbs (80%). Four patients (5.6%) developed meningoencephalitis, and three of them eventually died. In conclusion, considering its clinical and epidemiological characteristics, cutaneous anthrax must be included in the differential diagnosis of skin ulcers. A patient with clinical suspicion of the disease should receive effective treatment soon, in order to avoid neurological complications which carry a high fatality rate.

EVALUATION OF ANTIMICROBIAL AND CYTOTOXIC ACTIVITIES OF PLANT EXTRACTS FROM SOUTHERN MINAS GERAIS CERRADO

The antimicrobial activity of plant hidroethanolic extracts on bacteria Gram positive, Gram negative, yeasts, Mycobacterium tuberculosis H37 and Mycobacterium bovis was evaluated by using the technique of Agar diffusion and microdilution in broth. Among the extracts evaluated by Agar diffusion, the extract of Bidens pilosa leaf presented the most expressive average of haloes of growth inhibition to the microorganisms, followed by the extract of B. pilosa flower, of Eugenia pyriformis' leaf and seed, of Plinia cauliflora leaf which statistically presented the same average of haloes inhibitory formation on bacteria Gram positive, Gram negative and yeasts. The extracts of Heliconia rostrata did not present activity. Mycobacterium tuberculosis H37 and Mycobacterium bovis(BCG) appeared resistant to all the extracts. The susceptibility profile of Candida albicans and Saccharomyces cerevisiae fungi were compared to one another and to the Gram positive Bacillus subtilis, Enterococcus faecalis and the Gram negative Salmonella typhimurium bacteria (p > 0.05). The evaluation of cytotoxicity was carried out on C6-36 larvae cells of the Aedes albopictus mosquito. The extracts of stem and flower of Heliconia rostrata, leaf and stem of Plinia cauliflora, seed of Anonna crassiflora and stem, flower and root of B. pilosa did not present toxicity in the analyzed concentrations. The highest rates of selectivity appeared in the extracts of stem of A. crassiflora and flower of B. pilosa to Staphylococcus aureus, presenting potential for future studies about a new drug development.