175 resultados para Corynebacterium pseudotuberculosis


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The diphtheria tox repressor (DtxR) is a transition metal ion-dependent regulatory element that controls the expression of diphtheria toxin and several genes involved in the synthesis of siderophores in Corynebacterium diphtheriae. In the presence of transition metal ions apo-DtxR becomes activated and specifically binds to its target DNA sequences. We demonstrate by glutaraldehyde cross-linking that monomeric apo-DtxR is in weak equilibrium with a dimeric form and that upon addition of activating metal ions to the reaction mixture a dimeric complex is stabilized. Addition of the DNA-binding-defective mutant apo-DtxR(delta 1-47) to apo-DtxR in the absence of transition metal ions inhibits conversion of the apo-repressor to its activated DNA-binding form. We also show that the binding of Ni2+ to both apo-DtxR and apo-DtxR(delta 1-47) is cooperative and that upon ion binding there is a conformational change in the environment of the indole ring moiety of Trp-104. For the wild-type repressor the consequences of this conformational change include a shift in equilibrium toward dimer formation and activation of target DNA binding by the repressor. We conclude that the formation of DtxR homodimers is mediated through a protein-protein interaction domain that is also activated on metal ion binding.

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Este trabalho consiste em um estudo sobre monitoramento ambiental de áreas limpas. Os parâmetros comparados foram partículas não viáveis de 0,5 e 5,0 µm, contaminação do ambiente (UFC do ar) e de superfícies (Rodac®). Procedeu-se ao estudo em áreas de manipulação crítica (Classe A). Nestas áreas, a amostragem foi feita em situações de repouso e dinâmica, antes e após sanitização, e em etapas de certificação e rotina. Os dados de literatura indicam que os parâmetros não são particularmente dependentes do lay out ou classificação das áreas, mas sim do seu uso e do comportamento dos operadores. As conclusões foram positivas quanto a correlação entre diferentes locais de amostragem, para partículas não viáveis de 0,5 e 5,0 µm, e ausência deste tipo de correlação em posições em fluxo laminar. Também, os valores de correlação foram quase sempre decrescentes com a maior limpeza do ambiente. Os microrganismos mais frequentemente isolados, nas áreas A2., A3 e A4 foram Bacillus sp, Staphylococcus sp e Corynebacterium sp.

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A resistência de cinco microrganismos presentes na microbiota da área de produção estéril (Cristalização Estéril), frente a ação do gás de peróxido de hidrogênio foi determinada e o valor O obtido para cada microrganismo foi comparado ao valor D do Bacillus stearothermophilus ATCC 12980 exposto ao mesmo agente. Os microrganismos testados foram Bacillus sp, M. luteus, Corynebacterium, Staphylococcus sp e Penicillium sp. Este teste tinha a finalidade de comprovar que a resistência do Bacillus stearothermophilus é maior quando da exposição ao peróxido de hidrogênio se comparada a outros microrganismos presentes na área produtiva. A metodologia consistiu da inoculação de 0,01 mL da suspensão de cada microrganismo na contagem de 102UFC/0,01 mL em cupons de aço inoxidável, previamente esterilizados por calor seco e posterior exposição ao gás de peróxido de hidrogênio. O experimento demonstrou que o valor D obtido para o Bacillus stearothermophilus ésuperior aos obtidos para os outros microrganismos em teste comprovando que a escolha deste microrganismo para o desafio contra o peróxido de hidrogênio é apropriada. Também executou-se o teste que visava garantir que o aço inoxidável é o material de suporte mais recomendado para este fim, utilizando-se suportes de diversos materiais normalmente encontrados no interior dos isoladores (PVC, aço inoxidável, CKC, teflon, polipropileno, látex, silicone, Hypalon, vidro, nylon, saco de alumínio) com 0,01 mL de inóculo de Bacillus stearothermophilus na contagem de 102UFC/O,01 mL, o que foi devidamente comprovado.

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Inaug.-diss. - Hannover, 1913.

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The locus of enterocyte effacement (LEE) is a large multigene chromosomal segment encoding gene products responsible for the generation of attaching and effacing lesions in many diarrheagenic Escherichia coli strains. A recently sequenced LEE harboring a pathogenicity island (PAI) from a Shiga toxin E. coli serotype 026 strain revealed a LEE PAI (designated LEE 026) almost identical to that obtained from a rabbit-specific enteropathogenic 015:H- strain. LEE 026 comprises 59,540 bp and is inserted at 94 min within the mature pheU tRNA locus. The LEE 026 PAI is flanked by two direct repeats of 137 and 136 bp (DR1 and DR2), as well as a gene encoding an integrase belonging to the P4 integrase family. We examined LEE 026 for horizontal gene transfer. By generating mini-LEE plasmids harboring only DR1 or DR2 with or without the integrase-like gene, we devised a simple assay to examine recombination processes between these sequences. Recombination was shown to be integrase dependent in a Delta recA E. coli K-12 strain background. Recombinant plasmids harboring a single direct repeat cloned either with or without the LEE 026 integrase gene were found to insert within the chromosomal pheU locus of E. coli K-12 strains with equal efficiency, suggesting that an endogenous P4-like integrase can substitute for this activity. An integrase with strong homology to the LEE 026 integrase was detected on the K-12 chromosome associated with the leuX tRNA locus at 97 min. Strains deleted for this integrase demonstrated a reduction in the insertion frequency of plasmids harboring only the DR into the pheU locus. These results provide strong evidence that LEE-harboring elements are indeed mobile and suggest that closely related integrases present on the chromosome of E. coli strains contribute to the dynamics of PAI mobility.

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The surface microflora (902 isolates) of Livarot cheeses from three dairies was investigated during ripening. Yeasts were mainly identified by Fourier transform infrared spectroscopy. Geotrichum candidum was the dominating yeast among 10 species. Bacteria were identified using Biotype 100 strips, dereplicated by repetitive extragenic palindromic PCR (rep-PCR); 156 representative strains were identified by either BOX-PCR or (GTG) 55-PCR, and when appropriate by 16S rDNA sequencing and SDS-PAGE analysis. Gram-positive bacteria accounted for 65% of the isolates and were mainly assigned to the genera Arthrobacter, Brevibacterium, Corynebacterium, and Staphylococcus. New taxa related to the genera Agrococcus and Leucobacter were found. Yeast and Gram-positive bacteria strains deliberately added as smearing agents were sometimes undetected during ripening. Thirty-two percent of the isolates were Gram-negative bacteria, which showed a high level of diversity and mainly included members of the genera Alcaligenes, Hafnia, Proteus, Pseudomonas, and Psychrobacter. Whatever the milk used (pasteurized or unpasteurized), similar levels of biodiversity were observed in the three dairies, all of which had efficient cleaning procedures and good manufacturing practices. It appears that some of the Gramnegative bacteria identified should now be regarded as potentially useful in some cheese technologies. The assessment of their positive versus negative role should be objectively examined.

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Comprehensive collaborative studies from our laboratories reveal the extensive biodiversity of the microflora of the surfaces of smear-ripened cheeses. Two thousand five hundred ninety-seven strains of bacteria and 2,446 strains of yeasts from the surface of the smear-ripened cheeses Limburger, Reblochon, Livarot, Tilsit, and Gubbeen, isolated at three or four times during ripening, were identified; 55 species of bacteria and 30 species of yeast were found. The microfloras of the five cheeses showed many similarities but also many differences and interbatch variation. Very few of the commercial smear microorganisms, deliberately inoculated onto the cheese surface, were reisolated and then mainly from the initial stages of ripening, implying that smear cheese production units must have an adventitious "house" flora. Limburger cheese had the simplest microflora, containing two yeasts, Debaryomyces hansenii and Geotrichum candidum, and two bacteria, Arthrobacter arilaitensis and Brevibacterium aurantiacum. The microflora of Livarot was the most complicated, comprising 10 yeasts and 38 bacteria, including many gram-negative organisms. Reblochon also had a very diverse microflora containing 8 yeasts and 13 bacteria (excluding gram-negative organisms which were not identified), while Gubbeen had 7 yeasts and 18 bacteria and Tilsit had 5 yeasts and 9 bacteria. D. hansenii was by far the dominant yeast, followed in order by G. candidum, Candida catenulata, and Kluyveromyces lactis. B. aurantiacum was the dominant bacterium and was found in every batch of the 5 cheeses. The next most common bacteria, in order, were Staphylococcus saprophyticus, A. arilaitensis, Corynebacterium casei, Corynebacterium variabile, and Microbacterium gubbeenense. S. saprophyticus was mainly found in Gubbeen, and A. arilaitensis was found in all cheeses but not in every batch. C. casei was found in most batches of Reblochon, Livarot, Tilsit, and Gubbeen. C. variabile was found in all batches of Gubbeen and Reblochon but in only one batch of Tilsit and in no batch of Limburger or Livarot. Other bacteria were isolated in low numbers from each of the cheeses, suggesting that each of the 5 cheeses has a unique microflora. In Gubbeen cheese, several different strains of the dominant bacteria were present, as determined by pulsed-field gel electrophoresis, and many of the less common bacteria were present as single clones. The culture-independent method, denaturing gradient gel electrophoresis, resulted in identification of several bacteria which were not found by the culture-dependent (isolation and rep-PCR identification) method. It was thus a useful complementary technique to identify other bacteria in the cheeses. The gross composition, the rate of increase in pH, and the indices of proteolysis were different in most of the cheeses.

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La mastitis infecciosa es una patología común durante la lactancia y constituye una de las primeras causas de destete precoz. Por tanto, debería ser considerada un problema de Salud Pública relevante, ya que priva a la pareja madre-hijo de los incuestionables beneficios que la lactancia proporciona. No obstante, la mastitis humana ha sido hasta la fecha una enfermedad subestimada e infradiagnosticada, ya que habitualmente sólo se consideran mastitis los casos agudos que cursan con una sintomatología evidente y su diagnóstico microbiológico no se realiza de forma rutinaria. La etiopatogenia de la mastitis se ha relacionado con un proceso de disbiosis en la glándula mamaria, que da lugar al sobrecrecimiento de ciertas especies presentes en la leche humana. Sin embargo, la ausencia de un diagnóstico microbiológico rutinario de la leche humana hace que los estudios microbiológicos sobre esta patología sean muy escasos y que se desconozca el papel que juegan muchos agentes etiológicos. En este trabajo, el análisis microbiológico de 1.849 muestras de leche materna procedentes de mujeres con mastitis ha revelado que el género Staphylococcus constituye el primer grupo microbiano implicado en esta patología, siendo Staphylococcus epidermidis la especie aislada con mayor frecuencia (91,56% de las muestras). La especie Staphylococcus aureus se detectó en el 29,74% de los casos. Los géneros Streptocococcus y Corynebacterium constituyeron, respectivamente, el segundo (70,20%) y tercer (16,60%) grupo microbiano con mayor prevalencia en este estudio. Estos resultados ponen de manifiesto que los estafilococos coagulasa-negativos, los estreptococos del grupo viridans y las corinebacterias, habitualmente considerados microorganismos comensales y subestimados como causa de mastitis humanas, tienen un papel relevante como agentes etiológicos de esta patología. Este hecho avala que el análisis microbiológico de la leche, identificando los agentes causales a nivel de especie, es el único medio posible para obtener un diagnóstico etiológico preciso y establecer un tratamiento eficaz para la mastitis...

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Introduction Infective endocarditis (IE) has been reported to mimic granulomatosis with polyangiitis (GPA) and to test positive to antineutrophil cytoplasmic antibodies (ANCA), which may lead to a misdiagnosis and inappropriate treatment. Case presentation We report a case of a 59-year-old man admitted for purpura, gangrenous digital infarcts and glomerulonephritis. The diagnosis of IE was initially considered on the basis of heart murmur and two positive haemocultures to corynebacterium. Ineffectiveness of antimicrobial therapy and further neurological and nasal manifestations supported the diagnosis of GPA. Conclusions IE should be ruled out before initiation of immunosuppressive treatment. If the disease progresses despite antimicrobial treatment, vascular diseases should be rapidly taken into account in differential diagnosis and treated early to avoid fatal complications.

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Mestrado em Engenharia Zootécnica - Produção Animal - Instituto Superior de Agronomia