Estudos sobre a esporulação de uma amostra de bacillus: IV - Outras evidências sobre a atividade do íon Mn[2+] na esporulação endotrófica

Foram feitas experimentações om o intuito de se buscar mais evidências sobre a participação do íon Mn[2+] no mecanismo esporogenético de uma amostra de Bacillus licheniformis. Quando as formas vegetativas desta bactéria eram depositadas em um meio mineral, carente de fonte de carbono utilizável, em conjunto com um agente seqüestrante de metais como EDTA, a esporulação endotrófica deixava de ocorrer. Entretanto, a esporulação pôde ser protegida quando as células eram previamente saturadas com um excesso de Mn[2+] exógeno. As formas esporuladas obtidas nas condições estudadas mostraram termorresistência a 85ºC durante 20 minutos.

Controle em pós-colheita de Penicillium digitatum EM laranja-pera com microrganismos e tratamento térmico

O mercado consumidor está exigindo alimentos sem a presença de resíduos de agrotóxicos. Assim, o trabalho teve como objetivo avaliar o controle do bolor verde, em laranjas-Pera, com agentes de biocontrole (Bacillus subtilis e Bacillus licheniformis, Bacillus subtilis (QST 713)), associados ou não ao tratamento térmico. Para tanto, os frutos foram adquiridos em "packinghouse" antes do processamento, sendo lavados e desinfestados com hipoclorito de sódio. Os frutos submetidos a esses tratamentos foram armazenados, por 11 a 28 dias, em temperatura de 10 ºC e UR 90%±5 ou por oito dias a 20 ºC e UR 90%±5. De modo geral, o tratamento térmico reduziu a severidade da doença determinada pela área abaixo da curva do progresso da doença nos frutos e a incidência natural de doenças em pós-colheita de laranja-Pera. Por outro lado, os agentes de biocontrole não controlaram a doença, mostrando que os organismos testados não apresentaram atividade curativa contra o bolor verde.

Efeito de parâmetros hidrolíticos na obtenção de hidrolisados proteicos de farinha de trigo com baixo teor de fenilalanina

Tendo como objetivo a obtenção de hidrolisados proteicos de farinha de trigo com baixo teor de fenilalanina (Phe), foram preparados, inicialmente, extratos proteicos da farinha de trigo, empregando-se método enzimático pela ação de protease de Bacillus licheniformis. Em seguida, esses extratos foram hidrolisados sob a ação do extrato enzimático bruto (EEB), obtido de casca de abacaxi, e de pancreatina comercial; e alguns parâmetros hidrolíticos foram avaliados, tais como temperatura (30; 35; 40; 50; e 70 °C), tempo (1 hora e 30 minutos; 2 horas e 30 minutos; 3 horas e 30 minutos), e pH de reação (6,0; 7,0; 8,0 e 9,0). Para a remoção de Phe, empregou-se o carvão ativado (CA) e a eficiência deste processo foi avaliada determinando-se o teor de Phe por espectrofotometria derivada segunda, na farinha de trigo, assim como nos hidrolisados após tratamento com CA. Para os três parâmetros estudados, observaram-se efeitos variados sobre a remoção de Phe, sendo que os melhores resultados foram encontrados ao se empregar a associação sucessiva de EEB (E:S 10:100, 1 hora e 30 minutos), com a pancreatina (E:S 4:100, 3 horas e 30 minutos), em pH 7,0 a 50 °C, tendo atingido 66,28% de remoção de Phe, o que corresponde a um teor final de Phe de 522,44 mg.100 g-1 de hidrolisado.

Sugarcane starch: quantitative determination and characterization

Starch is found in sugarcane as a storage polysaccharide. Starch concentrations vary widely depending on the country, variety, developmental stage, and growth conditions. The purpose of this study was to determine the starch content in different varieties of sugarcane, between May and November 2007, and some characteristics of sugarcane starch such as structure and granules size; gelatinization temperature; starch solution filterability; and susceptibility to glucoamylase, pullulanase, and commercial bacterial and fungal α-amylase enzymes. Susceptibility to debranching amylolytic isoamylase enzyme from Flavobacterium sp. was also tested. Sugarcane starch had spherical shape with a diameter of 1-3 µm. Sugarcane starch formed complexes with iodine, which showed greater absorption in the range of 540 to 620 nm. Sugarcane starch showed higher susceptibility to glucoamylase compared to that of waxy maize, cassava, and potato starch. Sugarcane starch also showed susceptibility to debranching amylolytic pullulanases similar to that of waxy rice starch. It also showed susceptibility to α-amylase from Bacillus subtilis, Bacillus licheniformis, and Aspergillus oryzae similar to that of the other tested starches producing glucose, maltose, maltotriose, maltotetraose, maltopentose and limit α- dextrin.

Trypsin Inhibitor from Edible Mushroom Pleurotus floridanus Active against Proteases of Microbial Origin

Protease inhibitors can be versatile tools mainly in the fields of medicine, agriculture and food preservative applications. Fungi have been recognized as sources of protease inhibitors, although there are only few such reports on mushrooms. This work reports the purification and characterization of a trypsin inhibitor from the fruiting body of edible mushroom Pleurotus floridanus (PfTI) and its effect on the activity of microbial proteases. The protease inhibitor was purified up to 35-fold by DEAE-Sepharose ion exchange column, trypsin-Sepharose column and Sephadex G100 column. The isoelectric point of the inhibitor was 4.4, and its molecular mass was calculated as 37 kDa by SDS-PAGE and 38.3 kDa by MALDI-TOF. Inhibitory activity confirmation was by dot-blot analysis and zymographic activity staining. The specificity of the inhibitor toward trypsin was with Ki of 1.043×10−10 M. The inhibitor was thermostable up to 90 °C with maximal stability at 30 °C, active over a pH range of 4–10 against proteases from Aspergillus oryzae, Bacillus licheniformis, Bacillus sp. and Bacillus amyloliquefaciens. Results indicate the possibility of utilization of protease inhibitor from P. floridanus against serine proteases

Recognition of greater diversity of Bacillus species and related bacteria in human faeces

In a study looking at the culturable, aerobic Actinobacteria associated with the human gastrointestinal tract, the vast majority of isolates obtained from dried human faeces belonged to the genus Bacillus and related bacteria. A total of 124 isolates were recovered from the faeces of 10 healthy adult donors. 16S rRNA gene sequence analyses showed the majority belonged to the families Bacillaceae (n = 81) and Paenibacillaceae (n = 3), with Bacillus species isolated from all donors. Isolates tentatively identified as Bacillus clausii (n = 32) and B. licheniformis (n = 28) were recovered most frequently, with the genera Lysinibacillus, Ureibacillus, Oceanobacillus, Ornithinibacillus and Virgibacillus represented in some donors. Phenotypic data confirmed the identities of isolates belonging to well-characterized species. Representatives of the phylum Actinobacteria were recovered in much lower numbers (n = 11). Many of the bacilli exhibited antimicrobial activity against one or more strains of Clostridium difficile, C. perfringens, Listeria monocytogenes and Staphylococcus aureus, with some (n = 12) found to have no detectable cytopathic effect on HEp-2 cells. This study has revealed greater diversity within gut-associated aerobic spore-formers than previous studies, and suggests that bacilli with potential as probiotics could be isolated from the human gut.

Screening for Bacillus isolates in the broiler gastrointestinal tract

Spores from a number of different Bacillus species are currently being used as human and animal probiotics, although their mechanisms of action remain poorly understood. Here we describe the isolation of 237 presumptive gut-associated Bacillus spp. isolates that were obtained by heat and ethanol treatment of fecal material from organically reared broilers followed by aerobic plating. Thirty-one representative isolates were characterized according to their morphological, physiological, and biochemical properties as well as partial 16S rRNA gene sequences and screening for the presence of plasmid DNA. The Bacillus species identified included B. subtilis, B. pumilus, B. licheniformis, B. clausii, B. megaterium, B. firmus, and species of the B. cereus group, whereas a number of our isolates could not be classified. Intrinsic properties of potential importance for survival in the gut that could be advantageous for spore-forming probiotics were further investigated for seven isolates belonging to five different species. All isolates sporulated efficiently in the laboratory, and the resulting spores were tolerant to simulated gastrointestinal tract conditions. They also exhibited antimicrobial activity against a broad spectrum of bacteria, including food spoilage and pathogenic organisms such as Bacillus spp., Clostridium perfringens, Staphylococcus aureus, and Listeria monocytogenes. Importantly, the isolates were susceptible to most of the antibiotics tested, arguing that they would not act as donors for resistance determinants if introduced in the form of probiotic preparations. Together, our results suggest that some of the sporeformers isolated in this study have the potential to persist in or transiently associate with the complex gut ecosystem.

Lipopeptides Produced by a Soil Bacillus Megaterium Strain

A soil microorganism identified as Bacillum megaterium was found to produce several antibiotics substances after growth for 20 h at 37A degrees C in a mineral culture medium. Analysis both by electron spray ionization (ESI) and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) identified these substances as lipopeptides. Predominant peaks at m/z 1,041 and m/z 1,065 revealed ions which are compatible with surfactins and lichenysins, respectively. Two other ions m/z 1,057 and m/z 1,464 were further studied by collision-induced dissociation (CID) unveiling an iturin A at the first and fengycins A and B at the second m/z peaks. The CID spectrum of the m/z 1,464 ion also suggests the existence of fengycins A and B variants in which Ile was changed to Val in the position 10 of the peptide moiety. Raw mixtures of all these compounds were also assayed for antibiotic features. The data enlighten the unusual diversity of the lipopeptide mixture produced by a sole Bacillus species.

Amylase utilization for the extrusion of dog diets

Effects of amylase addition on extruder parameters, cost of extrusion, kibble quality and digestibility of dog food were measured in two separate experiments. In experiment 1, 120 kilo-novo-alpha-amilase-unit (KNU)/kg of heat stable alpha-amylase produced by Bacillus licheniformis was added in liquid form during a preconditioning period. In experiment 23684 KNU/kg of heat stable alpha-amylase produced by Aspergillus oryzae was mixed with the ingredients before extrusion. The diets were processed in a single screw extruder and submitted to digestibility and on experiment 1 also to palatability tests. Digestibility was tested using 12 dogs, six per diet. Data were submitted to analysis of variance followed by F-test. Amylase addition altered extrusion parameters in both experiments (P<0.05), with higher output (kg of dry matter [DM]/h: 28% and 43% higher in experiments 1 and 2) and less electric energy consumption (kW to produce 100 kg DM: 22% and 29% lower in experiments 1 and 2). Kibble appearance and quality [density (g/L), cutting force (g), and starch gelatinization degree (%)] did not change with enzyme treatment (P>0.05). Likewise, enzyme addition did not change nutrient digestibility, fecal dry matter or food palatability (P<0.05). Taken together our results suggest that amylase promoted the breakdown of amylose chains, thereby reducing the dough viscosity and resistance inside the extruder which allowed for higher product flow and less electricity energy consumption without altering food quality. (C) 2012 Elsevier B.V. All rights reserved.

Controle em pós-colheita de Penicillium digitatum em laranja-pera com microrganismos e tratamento térmico

The consumer market demands food without pesticide residues. Therefore, this study focused on evaluating the control of green mold in Pera orange trees with biocontrol agents (Bacillus subtilis, Bacillus licheniformis and Bacillus subtilis (QST 713)), associated or not with heat treatment. The fruit was obtained in packinghouse before processing, being washed and disinfected with the use of Sodium Hypochlorite. Fruits submitted to these treatments were stored from 11 to 28 days at temperature of 10 °C and RH 90%±5 or for eight days at 20 °C and 90%±5. In general, the heat treatment reduced the disease severity determine by the area under the disease progress curve in the fruit and the incidence of natural postharvest disease in Pera oranges. On the other hand, biocontrol agents did not control the disease, showing that the organisms tested did not present curative activity against the green mold.

Probióticos para leitões lactentes e na fase de creche

Pós-graduação em Zootecnia - FMVZ

Influência da adição de enzimas e microrganismos sobre a digestão anaeróbia de dejetos bovinos e suínos

Pós-graduação em Zootecnia - FCAV

Controle em pós-colheita de bolor verde em laranja pêra com microrganismos e tratamento térmico

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Controle da ferrugem asiática da soja (Phakopsora packyrhizi) com óleo de café e bacillus spp

Pós-graduação em Agronomia (Proteção de Plantas) - FCA

Probiótico e enzimas em dietas de frangos de corte: desempenho, características da cama e excretas e produção de biogás

Pós-graduação em Zootecnia - FCAV