RESUMO

In this study, five bacteriocin-producing Lactococcus lactis strains were identified from different naturally fermented Brazilian sausages. Ion exchange and reversed-phase chromatographies were used to purify the bacteriocins from culture supernatant of the five strains. Mass spectrometry (MALDI-TOF/TOF) showed that the molecular masses of the bactericoins from L. lactis ID1.5, ID3.1, ID8.5, PD4.7, and PR3.1 were 3330.567 Da, 3330.514 Da, 3329.985 Da, 3329.561 Da, and 3329.591 Da, respectively. PCR product sequence analysis confirmed that the structural genes of bacteriocins produced by the five isolates are identical to the lantibiotic nisin Z. Optimal nisin Z production was achieved in tryptone and casein peptone, at pH 6.0 or 6.5. The most favorable temperatures for nisin Z production were 25°C and 30°C, and its production was better under aerobic than anaerobic condition. The type of carbon source appeared to be an important factor for nisin Z production. While sucrose was found to be the most efficient carbon source for nisin Z production by four L. lactis isolates, fructose was the best for one isolate. Lactose was also a good energy source for nisin Z production. Surprisingly, glucose was clearly the poorest carbon source for nisin Z production. The five isolates produced different amounts of the bacteriocin, L. lactis ID1.5 and ID8.5 isolates being the best nisin Z producers. DNA sequence analysis did not reveal any sequence differences in the nisZ and nisF promoter regions that could explain the differences in nisin Z production, suggesting that there should be other factors responsible for differential nisin Z production by the isolates.

RESUMO

Novel compounds and innovative methods are required considering that antibiotic resistance has reached a crisis point. In the study, two cell-bound antimicrobial compounds produced by Lactococcus lactis ID1.5 were isolated and partially characterized. Following purification by cationic exchange and a solid-phase C18 column, antimicrobial activity was recovered after three runs of RPC using 60% (v/v) and 100% (v/v) of 2-propanol for elution, suggesting that more than one antimicrobial compound were produced by L. lactis ID1.5, which were in this study called compounds AI and AII. The mass spectrum of AI and AII showed major intensity ions at m/z 1070.05 and 955.9 Da, respectively. The compound AI showed a spectrum of antimicrobial activity mainly against L. lactis species, while the organisms most sensitive to compound AII were Bacillus subtilis, Listeria innocua, Streptococcus pneumoniae and Pseudomonas aeruginosa. The antimicrobial activity of both compounds was suppressed by treatment with Tween 80. Nevertheless, both compounds showed high stability to heat and proteases treatments. The isolated compounds, AI and AII, showed distinct properties from other antimicrobial substances already reported as produced by L. lactis, and have a significant inhibitory effect against two clinically important respiratory pathogens.

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The common bean is one of the most important legumes in the human diet, but little is known about the endophytic bacteria associated with the leaves of this plant. The objective of this study was to characterize the culturable endophytic bacteria of common bean (Phaseolus vulgaris. leaves from three different cultivars (Vermelhinho, Talismã, and Ouro Negro) grown under the same field conditions. The density of endophytic populations varied from 4.5 x 10² to 2.8 x 10³ CFU g-1 of fresh weight. Of the 158 total isolates, 36.7% belonged to the Proteobacteria, 32.9% to Firmicutes, 29.7% to Actinobacteria, and 0.6% to Bacteroidetes. The three P. vulgaris cultivars showed class distribution differences among Actinobacteria, Alphaproteobacteria and Bacilli. Based on 16S rDNA sequences, 23 different genera were isolated comprising bacteria commonly associated with soil and plants. The genera Bacillus, Delftia, Methylobacterium, Microbacterium, Paenibacillus, Staphylococcus and Stenotrophomonas were isolated from all three cultivars. To access and compare the community structure, diversity indices were calculated. The isolates from the Talismã cultivar were less diverse than the isolates derived from the other two cultivars. The results of this work indicate that the cultivar of the plant may contribute to the structure of the endophytic community associated with the common bean. This is the first report of endophytic bacteria from the leaves of P. vulgaris cultivars. Future studies will determine the potential application of these isolates in biological control, growth promotion and enzyme production for biotechnology.(AU)

Assuntos

RESUMO

The common bean is one of the most important legumes in the human diet, but little is known about the endophytic bacteria associated with the leaves of this plant. The objective of this study was to characterize the culturable endophytic bacteria of common bean (Phaseolus vulgaris. leaves from three different cultivars (Vermelhinho, Talismã, and Ouro Negro) grown under the same field conditions. The density of endophytic populations varied from 4.5 x 10² to 2.8 x 10³ CFU g-1 of fresh weight. Of the 158 total isolates, 36.7% belonged to the Proteobacteria, 32.9% to Firmicutes, 29.7% to Actinobacteria, and 0.6% to Bacteroidetes. The three P. vulgaris cultivars showed class distribution differences among Actinobacteria, Alphaproteobacteria and Bacilli. Based on 16S rDNA sequences, 23 different genera were isolated comprising bacteria commonly associated with soil and plants. The genera Bacillus, Delftia, Methylobacterium, Microbacterium, Paenibacillus, Staphylococcus and Stenotrophomonas were isolated from all three cultivars. To access and compare the community structure, diversity indices were calculated. The isolates from the Talismã cultivar were less diverse than the isolates derived from the other two cultivars. The results of this work indicate that the cultivar of the plant may contribute to the structure of the endophytic community associated with the common bean. This is the first report of endophytic bacteria from the leaves of P. vulgaris cultivars. Future studies will determine the potential application of these isolates in biological control, growth promotion and enzyme production for biotechnology.

Assuntos

RESUMO

The common bean is one of the most important legumes in the human diet, but little is known about the endophytic bacteria associated with the leaves of this plant. The objective of this study was to characterize the culturable endophytic bacteria of common bean (Phaseolus vulgaris) leaves from three different cultivars (Vermelhinho, Talismã, and Ouro Negro) grown under the same field conditions. The density of endophytic populations varied from 4.5 x 10(2) to 2.8 x 10(3) CFU g(-1) of fresh weight. Of the 158 total isolates, 36.7% belonged to the Proteobacteria, 32.9% to Firmicutes, 29.7% to Actinobacteria, and 0.6% to Bacteroidetes. The three P. vulgaris cultivars showed class distribution differences among Actinobacteria, Alphaproteobacteria and Bacilli. Based on 16S rDNA sequences, 23 different genera were isolated comprising bacteria commonly associated with soil and plants. The genera Bacillus, Delftia, Methylobacterium, Microbacterium, Paenibacillus, Staphylococcus and Stenotrophomonas were isolated from all three cultivars. To access and compare the community structure, diversity indices were calculated. The isolates from the Talismã cultivar were less diverse than the isolates derived from the other two cultivars. The results of this work indicate that the cultivar of the plant may contribute to the structure of the endophytic community associated with the common bean. This is the first report of endophytic bacteria from the leaves of P. vulgaris cultivars. Future studies will determine the potential application of these isolates in biological control, growth promotion and enzyme production for biotechnology.

RESUMO

Objetivo: Avaliar o uso do Lactobacillus delbruechii UFV H2B20 na prevenção de diarréia em crianças hospitalizadas. Método: Trata-se de estudo prospectivo, randomizado e duplo cego, no qual foram incluidas 139 crianças hospitalizadas com idades de 1 a 36 meses em enfermaria geral do Hospital Infantil João Paulo II/FHEMIG, em Belo Horizonte, no período de agosto de 2004 a julho de 2006...

Objectives: The objective of this trial was to evaluate the use of lactobacillus delbrueckii UFV H2B20 in the prevention of diarrhea in hospitalized children. Methods: It is a prospective, randomized and double blind trial in which were included 139 children from 1 to 36 months old who were hospitalized in the João Paulo II/FHEMIG children's Hospital, in Belo Horizonte, from August 2004 to July 2006...

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The growth of Salmonella enterica subs. enterica sorovar Typhimurium at 25ºC was monitored in industrialized and hospital formulated enteral feeds and the results were used to validate the mathematical model of Salmonella growth presented by the Pathogen Modeling Program (PMP) 7.0 (USDA-USA). The generation time of Salmonella in enteral feeds ranged from 21 to 34.8 min and, the maximum growth rate (µmax) varied from 1.28 to 1.95 h-1, resulting in a population increase from 5 to 6 log10 cycles within 14 to 24 h incubation. Growth was faster in the hospital formulated feed containing vegetables and eggs. The growth kinetic's parameters as lag phase; µmax and maximum population density (MPD) were similar to those predicted by the PMP 7.0, with exception of lag phase in enteral diet at pH 6.3. The results of this study validated the PMP 7.0 model for describe Salmonella growth in enteral feeds and demonstrates the appropriateness of use such model to determine the pathogen behavior in a wide range of storage conditions in this food.

O crescimento de Salmonella enterica subs. enterica sorovar Typhimurium a 25ºC foi determinado em dietas enterais industrializadas e formuladas em hospital e os resultados obtidos foram usados para validar um modelo matemático de crescimento de Salmonella apresentado no Programa de Modelagem de Patógenos (PMP), versão 7,0 (USDA-EUA). O tempo de geração de Salmonella em dietas enterais variou de 21 a 34,8 min e a velocidade específica máxima de crescimento (µmax) foi de 1,28 a 1,95 h-1, resultando em aumento de 5 a 6 ciclos logarítimos em um período de 14 a 24 h de incubação. O crescimento foi mais rápido na dieta formulada em hospital contendo vegetais e ovos. Os parâmetros cinéticos como fase lag, µmax e densidade populacional máxima (MDP) foram similares aqueles previstos no PMP 7.0, com exceção da fase lag em dietas enteral com pH 6,3. Os resultados deste estudo validaram o modelo do PMP 7,0 para descrever o crescimento de Salmonella em dietas enterais e demonstraram a propriedade desse modelo para determinar o comportamento do patógeno em uma variedade de condições nesse tipo de alimento.

Assuntos

RESUMO

The growth of Salmonella enterica subs. enterica sorovar Typhimurium at 25ºC was monitored in industrialized and hospital formulated enteral feeds and the results were used to validate the mathematical model of Salmonella growth presented by the Pathogen Modeling Program (PMP) 7.0 (USDA-USA). The generation time of Salmonella in enteral feeds ranged from 21 to 34.8 min and, the maximum growth rate (µmax) varied from 1.28 to 1.95 h(-1), resulting in a population increase from 5 to 6 log10 cycles within 14 to 24 h incubation. Growth was faster in the hospital formulated feed containing vegetables and eggs. The growth kinetic's parameters as lag phase; µmax and maximum population density (MPD) were similar to those predicted by the PMP 7.0, with exception of lag phase in enteral diet at pH 6.3. The results of this study validated the PMP 7.0 model for describe Salmonella growth in enteral feeds and demonstrates the appropriateness of use such model to determine the pathogen behavior in a wide range of storage conditions in this food.

RESUMO

To obtain in-depth information on the overall metabolic behavior of the new good xylitol producer Debaryomyces hansenii UFV-170, batch bioconversions were carried out using semisynthetic media with compositions simulating those of typical acidic hemicellulose hydrolysates of sugarcane bagasse. For this purpose, we used media containing glucose (4.3-6.5 g/L), xylose (60.1-92.1 g/L), or arabinose (5.9-9.2 g/L), or binary or ternary mixtures of them in either the presence or absence of typical inhibitors of acidic hydrolysates, such as furfural (1.0-5.0 g/L), hydroxymethylfurfural (0.01- 0.30 g/L), acetic acid (0.5-3.0 g/L), and vanillin (0.5-3.0 g/L). D. hansenii exhibited a good tolerance to high sugar concentrations as well as to the presence of inhibiting compounds in the fermentation media. It was able to produce xylitol only from xylose, arabitol from arabinose, and no glucitol from glucose. Arabinose metabolization was incomplete, while ethanol was mainly produced from glucose and, to a lesser less extent, from xylose and arabinose. The results suggest potential application of this strain in xyloseto- xylitol bioconversion from complex xylose media from lignocellulosic materials.

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Listeria monocytogenes is an opportunistic psychrotroph foodborne pathogen that has been used as a model organism to study the efficacy of many different preservation methods. This work aimed to test the antilisterial activity of lactic acid bacteria isolated from Italian salami and study the development of resistance. Isolates were obtained from naturally fermented Italian salami and cultures that retained activity in the supernatants after pH neutralization and catalase treatment were further characterized. The isolate showing highest inhibitory activity (PD 6.9) was tested for sensibility to proteases, heat and pH. To evaluate if resistance developed, sensitive strains were transferred with sub-lethal doses of the partially purified inhibitory substance and then inoculated into media containing higher doses of the extract. Isolate PD 6.9 inhibited several L. monocytogenes strains obtained from different origins and retained its activity over a wide range of pH and temperature. When increasing concentrations (10-100 AU ml(-1)) of the partially purified inhibitory substance were added to culture media, growth of L. monocytogenes did not occur even after 12 h of incubation. Cultures of Listeria that were transferred with sub-lethal doses (10 AU ml(-1)) of the partially purified inhibitory substance could resist higher doses of the extract (50 AU ml(-1)), but were inhibited when the concentration was further increased (100 AU ml(-1)). These results indicate that isolate PD 6.9 could potentially be used as a bioprotective culture for salami fermentation.

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RESUMO

Operons ribossomais têm sido instrumentos importantes na caracterizacão de comunidades microbianas e no estudo de relacionamentos entre microrganismos, principalmente em bactérias do ácido láctico. Operons ribossomais da linhagem probiótica, Lactobacillus delbrueckii UFV H2b20, foram parcialmente caracterizados. Um banco genômico da linhagem foi construído e os clones, contendo parte do operon ribossomal, foram subclonados pelo método de "shot gun", para em seguida serem seqüenciados com primer "forward". As seqüências indicaram a presenca da extremidade 3' do rDNA 16S seguida da região espacadora curta 1 (16S-23S) e a presenca da extremidade 3' do rDNA 23S seguido da região espacadora 2 (23S-5S), que por sua vez precedia o rDNA 5S. Adjacente ao gene rDNA 5S deste operon rrn uma região codificadora de 6 tRNAs foi detectada.

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RESUMO

Multidrug efflux mechanisms in bacteria contribute significantly to intrinsic and acquired resistance to antimicrobial agents. Genome analysis have confirmed the broad distribution of these systems in Gram-negative as well as in Gram-positive bacteria. Among resistance mechanisms, the multidrug efflux system or pump deserves special attention, since a cell that has acquired it can simultaneously diminish or even suppress the susceptibility to a wide range of antimicrobials. The efflux system is mediated by transport proteins which confer resistance to toxic compounds. In Gram-negative bacteria, a tripartite efflux system is necessary to expel the drug to the outer medium: a protein localized in the cytoplasmic membrane; another in the periplasmatic space (membrane fusion protein - MFP); and a third in the outer membrane (outer membrane factor - OMF). The drug transport is active, and depends either on the energy provided by ATP hydrolysis or is directly driven by the proton motive force. The transport proteins are grouped in families, according to the homology of the amino acid sequences and to similarity of mechanisms. Among Gram-negative bacteria, Escherichia coli and Pseudomonas aeruginosa have most of the hitherto identified and studied multidrug efflux systems.

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Bactérias láticas foram isoladas durante o processamento de salame tipo italiano, obtido a partir de duas plantas de processamento, no Estado do Paraná. Para o isolamento, foram utilizados os meios MRS, D-MRS e M17. Um total de 484 isolados teve sua atividade antibacteriana testada sobre Listeria monocytogenes (Laboratório de Microbiologia - UFV), Staphylococcus aureus (ATCC 25923), Salmonella enteritidis (ATCC 13076) e Escherichia coli (ATCC 25922), utilizando-se o deferred method. Cento e quinze isolados apresentaram zonas de inibição sobre, pelo menos, duas das bactérias indicadoras. Os isolados apresentaram maior inibição sobre Listeria monocytogenes. Os 45 isolados que apresentaram zonas de inibição sobre as quatro bactérias indicadoras foram identificados, utilizando-se o sistema Biolog. Lactobacillus bifermentans predominou entre os isolados identificados, obtidos no meio MRS, a partir das amostras das duas plantas de processamento.

RESUMO

Lactic acid bacteria were isolated in the MRS, D-MRS and M17 modified media, during the processing of Italian salami, obtained from two processing plants, in the State of Paraná. The 484 isolates were tested for their antibacterial activity against Listeria monocytogenes (Microbiology Laboratory-UFV), Staphylococcus aureus (ATCC 25923), Salmonella enteritidis (ATCC 13076) and Escherichia coli (ATCC 25922), by a deferred method. One hundred and fifteen isolates inhibited at least two of the pathogens. The isolates presented larger inhibition against L. monocytogenes. The 45 isolates with antagonistic action on the four indicator bacteria were identified by the system Biolog. Lactobacillus bifermentans prevailed among the isolates identified in MRS medium from the samples of the two processing plants.

Bactérias láticas foram isoladas durante o processamento de salame tipo italiano, obtido a partir de duas plantas de processamento, no Estado do Paraná. Para o isolamento, foram utilizados os meios MRS, D-MRS e M17. Um total de 484 isolados teve sua atividade antibacteriana testada sobre Listeria monocytogenes (Laboratório de Microbiologia - UFV), Staphylococcus aureus (ATCC 25923), Salmonella enteritidis (ATCC 13076) e Escherichia coli (ATCC 25922), utilizando-se o deferred method. Cento e quinze isolados apresentaram zonas de inibição sobre, pelo menos, duas das bactérias indicadoras. Os isolados apresentaram maior inibição sobre Listeria monocytogenes. Os 45 isolados que apresentaram zonas de inibição sobre as quatro bactérias indicadoras foram identificados, utilizando-se o sistema Biolog. Lactobacillus bifermentans predominou entre os isolados identificados, obtidos no meio MRS, a partir das amostras das duas plantas de processamento.