Halha Ostrensky Saridakis

Effects of medium composition, calcium, iron and oxygen on haemolysin production by Plesiomonas shigelloides isolated from water

Aims: The effects of medium composition, calcium, iron and oxygen tension on the haemolytic activity of Plesiomonas shigelloides were investigated.

Virulence properties of Escherichia coli strains belonging to enteropathogenic (EPEC) serogroups isolated from calves with diarrhea

Nineteen Escherichia coli strains belonging to enteropathogenic (EPEC) serogroups were isolated from calves with diarrhea in Paraná State. Brazil, and studied for virulence markers associated with EPEC or enterohemorrhagic E. coli (EHEC). The 19 isolates belonged to 12 serotypes with isolates of O26:H11, O119:H25 and O114:H- being the most prevalent Localized adherence (LA) was demonstrated for 37% of the isolates, consisting of all four O26:H11, both O114:H- and one O114:H40 isolates. All the LA strains were positive in the fluorescent-actin staining (FAS) test and possessed attaching-effacing E. coli (eae) sequences, but only O114 strains hybridized with the EPEC adherence factor (EAF) probe. None of the strains produced Shiga-like toxins (Verotoxin). Only the O26:H11 strains hybridized with the EHEC plasmid specific (CVD419) probe and were enterohemolytic, properties associated with EHEC strains. This investigation demonstrates that among the bovine strains isolated only those of serogroup O114 behaved as typical EPEC.

Prevalence and virulence properties of Vibrio cholerae non-O1, Aeromonas spp. and Plesiomonas shigelloides isolated from Cambé Stream (State of Paraná, Brazil)

The incidence of Vibrio cholerae, Aeromonas spp. and Plesiomonas shigelloides was determined in water samples from Cambé Stream. The samples were collected from seven different sites. The serogroups, virulence markers and drug resistance profiles were also evaluated. Twelve Aer. hydrophila, 12 Aer. caviae, eight Aer. sobria, seven Ple shigelloides and two V. cholerae non‐O1 were isolated. They belonged to different serogroups and all produced haemolysis in different assays. Five of the Aeromonas strains and one of V. cholerae non‐O1 were positive for enterotoxin activity. Haemagglutination and its inhibition, using erythrocytes of different origins, was variable for Aeromonas spp. and V. cholerae, while none of the Ple. shigelloides haemagglutinated in association with any type of erythrocyte. All isolates exhibited multiple drug resistance. These results indicate that the occurrence of V. cholerae non‐O1, Aeromonas spp. and Ple. shigelloides, in water used for vegetable irrigation, human recreation and animal consumption, among others, represents a potential risk for humans.

Prevalência da colonização vaginal e anorretal por estreptococo do grupo B em gestantes do terceiro trimestre

OBJETIVOS: estudar a prevalencia da colonizacao por estreptococo do grupo B em gestantes do terceiro trimestre, atendidas em um hospital regional, e avaliar a associacao da colonizacao com as variaveis maternas demograficas e clinicas. METODOS: no periodo de 8 de outubro de 2002 a 26 de marco de 2003, foi realizado um estudo transversal (de prevalencia) com 309 gestantes no terceiro trimestre. Amostras de secrecao vaginal e anorretal foram coletadas e testadas para identificacao presuntiva do estreptococo do grupo B. Foram incluidas as gestantes com gestacao maior ou igual a 36 semanas datadas por ultra-sonografia e excluidas as que se recusaram a participar, as em uso de antibioticoterapia e as que haviam sido submetidas a exame ginecologico pelo periodo minimo de 24 horas antes da coleta. As gestantes foram caracterizadas por variaveis demograficas (raca, idade, grau de escolaridade, renda familiar e numero de gestacoes) e clinicas (idade gestacional, ocorrencia de infeccao urinaria durante a gestacao atual, ruptura prematura de membranas e tempo de bolsa rota, febre materna intraparto, corioamnionite, liquido amniotico com meconio, via de parto utilizada, febre materna pos-parto e endometrite). RESULTADOS: das gestantes, 46 estavam colonizadas pelo estreptococo do grupo B, sendo que 26 (56,5%) tiveram a cultura vaginal positiva, 8 (17,4%) a cultura anorretal positiva e 12 (26,5%) tiveram tanto a cultura vaginal como a retal positivas. Nenhuma das variaveis analisadas neste estudo foi estatisticamente significativa quanto a colonizacao pelo estreptococo do grupo B. Os resultados obtidos foram submetidos a analise bivariada pelo teste do c2 e teste exato de Fisher quando apropriado. CONCLUSAO: a taxa de prevalencia da colonizacao vaginal e anorretal pelo estreptococo do grupo B em gestantes no terceiro trimestre, foi de 14,9%. Nao houve associacao entre fatores de risco (primigestacao, idade materna inferior a 20 anos e nivel socio economico baixo) e a prevalencia da infeccao.

Survival and conjugal transfer between Bacillus thuringiensis strains in aquatic environment

Field and laboratory studies were conducted to assess the survival of cells and spores and plasmid transfer between Bacillus thuringienis strains in aquatic environment. Results indicated that cells and spores of B. thuringiensis can survive for 10 days in water, without altering their number. The sporulation process began after 12-15 hours of inoculation of water. B. thuringiensis was able to transfer conjugative plasmids in the aquatic environment.

Antimicrobial activity and chemical investigation of Brazilian Drosera

The antimicrobial activity of three different extracts (hexanic, ethyl acetate, methanol) obtained from Brazilian Drosera species (D. communis, D. montana var. montana, D. brevifolia, D. villosa var. graomogolensis, D. villosa var. villosa, Drosera sp. 1, and Drosera sp. 2 ) were tested against Staphylococcus aureus (ATCC 25923), Enterococcus faecium (ATCC23212), Pseudomonas aeruginosa (ATCC27853), Escherichia coli (ATCC11229), Salmonella choleraesuis (ATCC10708), Klebsiella pneumoniae (ATCC13883), and Candida albicans (a human isolate). Better antimicrobial activity was observed with D. communis and D. montana var. montana ethyl acetate extracts. Phytochemical analyses from D. communis, D. montana var. montana and D. brevifolia yielded 5-hydroxy-2-methyl-1,4-naphthoquinone (plumbagin); long chain aliphatic hydrocarbons were isolated from D. communis and from D. villosa var. villosa, a mixture of long chain aliphatic alcohols and carboxylic acids, was isolated from D. communis and 3b-O-acetylaleuritolic acid from D. villosa var. villosa.

Isolation and partial characterization of a mutant of Bacillus thuringiensis producing melanin

A mutant (407-P) of Bacillus thuringiensis subsp. thuringiensis strain 407 producing a melanin was obtained after treatment with the mutagenic agent ethyl-methane-sulfonate. Several microbiological and biochemical properties of the two strains were analyzed and the results were similar. The mutant 407-P was also incorporated into non-sterilized soil samples, recovered, easily identified, and quantified, what enables its use in ecology of B. thuringiensis.

Pre-treatment with Concanavalin-A increases resistance of mice to peritoneal infection by Serratia marcescens

Mice pre-treated with Concanavalin-A largely survived an intra-peritoneal inoculum of 2 x 10(7) Serratia marcescens, whereas all control mice died within 15 h of inoculation. A subpopulation of peritoneal macrophages from Con-A pre-treated mice was able to phagocytose the bacteria in vitro (6.7 SEM 1.2% phagocytosing cells) and in vivo (16.9 SEM 2.1%), whereas control phagocytes did not phagocytose S. marcescens. The survival of Con-A pre-treated mice allowed their immunisation with living bacteria, and the antiserum thus produced increased the phagocytosis of S. marcescens in vitro. Control mice largely survived an inoculum of S. marcescens suspended in 50% immune serum, although the bacteria were resistant to the bactericidal activity of that serum. These results suggest that, in contrast to the delayed humoral protection afforded by immunisation, phagocytosis by phagocytes activated by Con-A conferred early protection to mice against experimental infection by S. marcescens.

A new method for asperphenamate synthesis and its antimicrobial activity evaluation

Asperphenamate is a natural phenylalanine derivative. This compound was produced through a new, two-step synthetic route. It was also evaluated by the antimicrobial activity of the pure substance against Escherichia coli, Staphylococcus aureus and Cladosporium herbarum.

Molecular and Phenotypic Characterization of Escherichia coli O26:H8 among Diarrheagenic E. coli O26 Strains Isolated in Brazil

ABSTRACT Escherichia coli strains of serogroup O26 comprise two distinct groups of pathogens, characterized as enteropathogenic E. coli (EPEC) and enterohemorrhagic E. coli (EHEC). Among the several genes related to type III secretion system-secreted effector proteins, espK was found to be highly specific for EHEC O26:H11 and its stx-negative derivative strains isolated in European countries. E. coli O26 strains isolated in Brazil from infant diarrhea, foods, and the environment have consistently been shown to lack stx genes and are thus considered atypical EPEC. However, no further information related to their genetic background is known. Therefore, in this study, we aimed to discriminate and characterize these Brazilian O26 stx-negative strains by phenotypic, genetic, and biochemical approaches. Among 44 isolates confirmed to be O26 isolates, most displayed flagellar antigen H11 or H32. Out of the 13 nonmotile isolates, 2 tested positive for fliC H11, and 11 were fliC H8 positive. The identification of genetic markers showed that several O26:H11 and all O26:H8 strains tested positive for espK and could therefore be discriminated as EHEC derivatives. The presence of H8 among EHEC O26 and its stx-negative derivative isolates is described for the first time. The interaction of three isolates with polarized Caco-2 cells and with intestinal biopsy specimen fragments ex vivo confirmed the ability of the O26 strains analyzed to cause attaching-and-effacing (A/E) lesions. The O26:H32 strains, isolated mostly from meat, were considered nonvirulent. Knowledge of the virulence content of stx-negative O26 isolates within the same serotype helped to avoid misclassification of isolates, which certainly has important implications for public health surveillance.