Isolamento de Vibrio parahaemolyticus e Vibrio cholerae em ostras, Crassostrea rhizophorae, coletadas em um criadouro natural no estuário do rio Cocó, Fortaleza, Ceará, Brasil

Oysters are edible organisms that are often ingested partially cooked or even raw, presenting therefore a very high risk to the consumers' health, especially in tropical regions. The presence of Vibrio cholerae and Vibrio parahaemolyticus in oysters sampled at an estuary in the Brazilian northeastern region was studied, with 300 oysters tested through an 8-months period. The salinity of the water at the sampling point varied between 3% and 27. V. cholerae was the most frequently detected species (33.3% of the samples), and of the 22 V. cholerae isolates, 20 were identified as non-O1/non-O139, with two of the colonies presenting a rough surface and most of remaining ones belonging to the Heiberg II fermentation group. V. parahaemolyticus was isolated from just one of the samples. Other bacteria such as Providencia spp., Klebsiella spp. and Morganella morganii were also isolated.As ostras são alimentos marinhos freqüentemente ingeridos crus ou parcialmente cozidos. Por esta razão, o risco para a saúde dos consumidores desses produtos é muito elevado, principalmente, quando são de regiões tropicais. Foi estudada a presença de Vibrio cholerae e Vibrio parahaemolyticus em ostras de um estuário na região Nordeste do Brasil. Trezentas ostras foram analisadas, em um período de 8 meses. A salinidade da água, no local de coleta, variou de 3 a 27. V. cholerae foi o vibrio mais freqüentemente detectado (33,3% das amostras). Dos 22 isolados, 20 foram identificados como V. cholerae não-O1/não-O139, duas delas apresentando forma rugosa sendo a maioria das demais pertencente ao tipo fermentativo Heiberg II. V. parahaemolyticus foi isolado em apenas umas das coletas. Foram, também, identificadas nas amostras isolados de Providencia spp., Klebsiella spp., Proteus spp. e Morganella morganii

Uso de extrato de plantas medicinais (Psidium guajava Linn. e Carica papaya Linn.) frente a bactérias isoladas de pescado, causadoras de diarréias infantis

Out of the twenty-four samples of shrimp and fish muscle used for this study, twelve were collected near a large marine sewer for waste disposal, 3 km off the coast of Fortaleza (Brazil) and used for the isolation of E. coli. Other twelve were collected at the Mucuripe fresh fish market (Fortaleza, Brazil) and used for the isolation of Staphylococcus aureus. Ethanol, water and acetone-diluted extracts of guava and papaya leaf sprouts were tested on the bacteria in order to verify their microbicidal potential. The E. coli strains used in the trials were rated LT positive. The papaya leaf extracts (Carica papaya Linn) showed no microbicidal activity while the guava sprout extracts (Psidium guajava Linn) displayed halos exceeding 13 mm for both species, an effect considered to be inhibitory by the method employed. Guava sprout extracts by 50% diluted ethanol most effectively inhibited E. coli (EPEC), while those in 50% acetone were less effective. It may be concluded that guava sprout extracts constitute a feasible treatment option for diarrhea caused by E. coli or by S. aureus-produced toxins, due to their quick curative action, easy availability in tropical countries and low cost to the consumer.Foram coletadas doze amostras de camarão e peixes nas imediações do interceptor oceânico, em Fortaleza e igual número na Feira de pescado do Mucuripe, Fortaleza, para isolamento de E. coli e Staphylococcus aureus, respectivamente. Extratos aquosos, alcoólicos e cetônicos de broto de goiabeira e de folha de mamão foram testados frente às bactérias para se verificar suas ações antibióticas. As cepas de E. coli utilizadas nos ensaios foram as classificadas como LT positivas. Os extratos de folhas de mamão (Carica papaya Linn) não revelaram quaisquer atividades antibióticas enquanto que os preparados com broto de goiabeira (Psidium guajava Linn) apresentaram halos sempre >;13 mm para as duas espécies, considerados como de inibição pelo método empregado. Os extratos de broto de goiabeira que apresentaram melhores resultados frente às cepas de E. coli ETEC foram os alcoólicos a 50% seguido do cetônico também a 50%. Concluímos que nos tratamentos de diarréias causadas por E. coli ou por toxinas elaboradas por S. aureus o extrato de brotos de goiabeira é uma opção devido a sua pronta ação curativa, seu fácil cultivo nos países tropicais e ao seu baixo valor aquisitivo

Phenotypic and genotypic characterization of bacteria Vibrio genus isolated in some estuaries of the State of CearÃ

Muitas pesquisas tÃm associado contaminaÃÃo aquÃtica ambiental com infecÃÃes de Vibrio em humanos, sugerindo que a importÃncia do monitoramento sistemÃtico das cepas ambientais se faz necessÃrio para definir seu possÃvel potencial patogÃnico e sua significÃncia clÃnica. O objetivo dessa pesquisa foi estudar a diversidade do gÃnero Vibrio isolado de quatro regiÃes estuarinas no Estado do CearÃ, (Pacoti, ChorÃ, Pirangi e Jaguaribe). As coletas realizadas resultaram num total de 32 amostras de Ãgua e 32 de sedimento, durante os meses de janeiro a abril de 2009. Foram catalogadas 19 espÃcies de bactÃrias pertencentes ao gÃnero Vibrio, das quais Vibrio parahaemolyticus e Vibrio alginolyticus foram as mais abundantes nos quatro estuÃrios: V. parahaemolyticus no Rio Chorà e V. alginolyticus no Rio Pacoti. As cepas identificadas foram submetidas a testes de susceptibilidade a quinze antimicrobianos. Todas as cepas analisadas (197) apresentaram susceptibilidade a sulfazotrim, ciprofloxacin, Ãcido nalidÃxico e cloranfenicol, sendo que cento e sessenta e trÃs (82%) apresentaram resistÃncia a penicilina G, cento e oito (54%) a ampicilina, quinze (7%) a cefalotina, trÃs (1%) a aztreonam, uma (0,5%) a gentamicina, a cefotaxima e a ceftriaxona. Cinquenta e uma cepas (25%) apresentaram comportamento intermediÃrio frente à cefalotina, vinte e oito cepas (14%) a ampicilina, dez (5%) a aztreonam, oito (4%) a tetracilina, duas (1%) a oxitetraciclina e uma (0,5%) a florfenicol, a cefotaxima, a ceftriaxona, a estreptomicina e a gentamicina. Foram escolhidas cinco espÃcies patÃgenas ao homem para verificaÃÃo de seus fatores de patogenicidade. As cepas identificadas como V. parahaemolyticus (64) e V. cholerae (9) foram analisadas atravÃs de tÃcnicas de biologia molecular, usando genes que confirmam as espÃcies e genes que indicam virulÃncia. Das 64 amostras de V. parahaemolyticus analisadas, 63 foram positivas para o gene tl, especÃfico para espÃcie, 57 para o gene tdh e 20 para o trh, genes que indicam patogenicidade. Das nove cepas de V. cholerae, cinco foram positivas para o gene ompW, gene especÃfico para espÃcie, porÃm, nenhuma amostra apresentou os genes de virulÃncia ctxA, zot, tcp e rfbO1. Com isso conclui-se que os estuÃrios dos rios analisados apresentam uma elevada abundÃncia de espÃcies, tendo V. parahaemolyticus e V. alginolyticus como as mais abundantes. O antibiograma das cepas isoladas mostrou uma elevada resistÃncia à penicilina e a ampicilina. Foram encontradas elevada positividade para a presenÃa dos fatores de virulÃncia nas cepas pertencentes Ãs espÃcies de Vibrio patÃgenas a humanos. As cepas de V. parahaemolyticus apresentaram genes de virulÃncia indicando que as cepas podem acarretar danos à saÃde pÃblica. A presenÃa do V. cholerae foi confirmada nas Ãguas e sedimento dos estuÃriosThe frequent association of environmental aquatic contamination with vibriosis in humans suggests the need for systematic monitoring and study of environmental vibrio strains and their pathogenic potential and clinical significance. The objective of this study was to evaluate the diversity of vibrio species in four estuaries (Pacoti, ChorÃ, Pirangi and Jaguaribe) in CearÃ, Northeastern Brazil. Nineteen vibrio species were identified in 32 water samples and 32 sediment samples collected between January and April 2009. Overall, V. parahaemolyticus and V. alginolyticus were the most abundant (the former in ChorÃ, the latter in Pacoti). The isolated strains were submitted to antibiogram testing with 15 antibiotics. All strains (n=197) were susceptible to sulfametoxazol-trimetoprim, ciprofloxacin, nalidixic acid and chloramphenicol. Resistance was observed to penicillin G (n=163; 82%), ampicillin (n=108; 54%), cephalothin (n=15; 7%), aztreonam (n=3; 1%), gentamicin, cefotaxime, ceftriaxone (1 each; 0.5%). Partial resistance was observed to cefalotin (n=52; 25%), ampicillin (n=28; 14%), aztreonam (n=10; 5%), tetracycline (n=8; 4%), oxytetracycline (n=2; 1%), and florfenicol, cefotaxime, ceftriaxone, streptomycin and gentamicin (1 each; 0.5%). Five species known to be pathogenic to humans were chosen for analysis of factors of pathogenicity. Strains belonging to the species V. parahaemolyticus (n=64) and V. cholerae (n=9) were submitted to molecular analysis using genes to confirm the species and indicate virulence. Sixty-three strains of V. parahaemolyticus were positive for species-specific tl, 57 were positive for tdh and 20 for trh. Five strains of V. cholerae were positive for species-specific ompW, but no strains presented the genes ctxA, zot, tcp or rfbO1. In conclusion, the estuaries surveyed presented a great diversity of vibrio species, the most abundant of which were V. parahaemolyticus and V. alginolyticus. Resistance to penicillin and ampicillin was elevated and positivity for virulence factors was considerable among strains of species pathogenic to humans. V. parahaemolyticus strains presented virulence genes indicating risk to public health. V. cholerae was identified in samples of both water and sedimen

<b>Ecotypes and virulence factors of <i>Salmonella</i> spp. detected in shrimp farms in State of Ceara, Brazil

  Our objective was to group in ecotypes 12 serovars of Salmonella isolated from shrimp farming environments in the State of Ceara (Northeast Brazil). Grouping was done based on genotypic virulence factors. Two groups based on the similarity of the Box-PCR were identified: a group consisting of three strains (01 S. ser. Madelia serovar and 02 S. ser. enterica subs. houtenae) and another group consisting of nine isolates (02 S. ser. Saintpaul serovars; 03 S. ser. Infantis; 02 S. ser. Panama; 01 S. enterica subs. enterica; and 01 S. enterica subs. houtenae). Distribution pattern of the serovars was not influenced by the origin matrices (water and sediment). Plasmid virulence genes pefA and invA were detected, unrelated to the serovar and environmental origin of the isolates. The presence of virulence genes in the isolates underlines the potential to trigger salmonellosis events via shrimp consumption. Biomonitoring of these sources of contamination should be encouraged as a protective measure, minimizing health risks and economic losses for the industry.

DETECTION OF VIRULENCE GENES IN ENVIRONMENTAL STRAINS OF Vibrio cholerae FROM ESTUARIES IN NORTHEASTERN BRAZIL

O objetivo deste estudo foi detectar a presença potencial virulência de Vibrio cholerae isolado de estuários do Nordeste do Brasil. Amostras de água e sedimento foram coletadas e inoculadas sobre meio seletivo para víbrios (TCBS) e colônias com características morfológicas de V. cholerae foram isoladas. A identificação fenotípica seguiu chave dicotômica baseada em caraterísticas bioquímicas. Foram empregadas as técnicas de amplificação da polimerase em cadeia (PCR) utilizando o gene ompW e a de multiplex PCR para detecção de genes de virulência (ctx, tcp, zot e rfbO1). Os resultados da identificação das diferentes abordagens foram comparados. Nove cepas de V. cholerae foram identificadas fenotipicamente e cinco confirmadas através da detecção do gene ompW. A chave dicotômica utilizada foi eficiente para a confirmação da espécie. Os quatro estuários analisados apresentaram estirpes de V. cholerae, e nenhuma das cepas isoladas apresentaram genes de virulência.The objectives of this study were to detect the presence of Vibrio cholerae in tropical estuaries (Northeastern Brazil) and to search for virulence factors in the environmental isolates. Water and sediment samples were inoculated onto a vibrio-selective medium (TCBS), and colonies with morphological resemblance to V. cholerae were isolated. The cultures were identified phenotypically using a dichotomous key based on biochemical characteristics. The total DNA extracted was amplified by PCR to detect ompW and by multiplex PCR to detect the virulence genes ctx, tcp, zot and rfbO1. The results of the phenotypic and genotypic identification were compared. Nine strains of V. cholerae were identified phenotypically, five of which were confirmed by detection of the species-specific gene ompW. The dichotomous key was efficient at differentiating environmental strains of V. cholerae. Strains of V. cholerae were found in all four estuaries, but none possessed virulence genes

Plectranthus amboinicus essential oil and carvacrol bioactive against planktonic and biofilm of oxacillin- and vancomycin-resistant Staphylococcus aureus

Abstract Background The emergence of multidrug-resistant bacteria is a worldwide concern and in order to find an alternative to this problem, the occurrence of antimicrobial compounds in Plectranthus amboinicus essential oil was investigated. Thus, this study aims to determine susceptibility of Staphylococcus aureus isolated from food to antibiotics, P. amboinicus essential oil (PAEO) and carvacrol. Methods Leaves and stem of P. amboinicus were used for extraction of essential oil (PAEO) by hydrodistillation technique and EO chemical analysis was performed by gas chromatography coupled to a mass spectrometer. S. aureus strains (n = 35) isolated from food and S. aureus ATCC 6538 were used to evaluate the antimicrobial and antibiofilm activity of PAEO and carvacrol. All strains (n = 35) were submitted to antimicrobial susceptibility profile by disk diffusion method. Determination of MIC and MBC was performed by microdilution technique and antibiofilm activity was determined by microtiter-plate technique with crystal violet assay and counting viable cells in Colony Forming Units (CFU). Results Carvacrol (88.17%) was the major component in the PAEO. Antibiotic resistance was detected in 28 S. aureus strains (80%) and 12 strains (34.3%) were oxacillin and vancomycin-resistant (OVRSA). From the 28 resistant strains, 7 (25%) showed resistance plasmid of 12,000 bp. All strains (n = 35) were sensitive to PAEO and carvacrol, with inhibition zones ranging from 16 to 38 mm and 23 to 42 mm, respectively. The lowest MIC (0.25 mg mL−1) and MBC (0.5 mg mL−1) values were observed when carvacrol was used against OVRSA. When a 0.5 mg mL−1 concentration of PAEO and carvacrol was used, no viable cells were found on S. aureus biofilm. Conclusion The antibacterial effect of carvacrol and PAEO proves to be a possible alternative against planktonic forms and staphylococcal biofilm

Complete genome sequence of native Bacillus cereus strains isolated from intestinal tract of the crab Ucides sp.

Bacillus cereus is a gram positive bacterium with sporulation capacity. Here, we report the complete genome sequence of two native B. cereus strains (#25 and #29) isolated from intestinal tract of the crab Ucides sp. from Pacoti River in the State of Ceará, Brazil. The findings of this study might increase the molecular information for Bacillus strains. The data can be used in comparative analyses, origin and distribution, as well support for genetic engineering. Keywords: Gram positive bacteria, Genome sequence, Bacillu