MALDI-TOF MS as a tool for the identification of Vibrio alginolyticus from Perna perna mussels (Linnaeus, 1758) / MALDI-TOF MS como ferramenta na identificação de Vibrio alginolyticus isolados de mexilhões Perna perna (Linnaeus, 1758)

Vibrio species are ubiquitous in aquatic environments, including coastal and marine habitats. Vibrio alginolyticus is an opportunistic pathogen for fish, crustaceans and mussels and their identification by biochemical tests may be impaired due their nutritional requirements. The study used Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) to identify 49 Vibrio spp. isolates associated with mussels (Perna perna) from different locations along the Rio de Janeiro coast. The rpoA gene was used as a genus-specific marker of Vibrio spp. and was positive in all 209 isolates. MALDI-TOF MS confirmed 87.8% of V. alginolyticus when compared to the results of the biochemical tests. Four isolates were identified as Shewanella putrefaciens (8.16%) and one was identified as V. parahaemolyticus (2.0%). Just one isolate was not identified by this technique (2.0%). The pyrH sequencing confirmed 75% of the proteomic technique results. MALDI-TOF MS is an excellent option for characterization of bacterial species, as it is efficient, fast and easy to apply. In addition, our study confirms its high specificity and sensitivity in these marine bacteria identification.(AU)

Espécies de Vibrio são ubiquitárias em ambientes aquáticos, incluindo habitats costeiros e marinhos. A espécie Vibrio alginolyticus é oportunista para peixes, crustáceos e moluscos e a sua identificação através de testes bioquímicos pode ter a qualidade prejudicada devido às suas exigências nutricionais. O presente estudo utilizou Espectrometria de Massa por Tempo de Vôo de Ionização/Desorção por Laser Assistida por Matriz (MALDI-TOF MS) para identificar diferentes espécies de Vibrio provenientes de mexilhões (Perna perna) coletados em diferentes locais ao longo da costa do Rio de Janeiro. O gene rpoA foi utilizado como um marcador gênero-específico de Vibrio spp. sendo positivo em todos os 209 isolados. MALDI-TOF MS confirmou 87,75% de V. alginolyticus quando comparados com os resultados dos testes bioquímicos. Quatro isolados foram identificados como Shewanella putrefaciens (8,16%), um como V. parahaemolyticus (2,0%) e apenas um (2,0%) não foi identificado pela técnica proteômica. E o sequenciamento do pyrH confirmou 75% dos resultados da técnica proteomica. MALDI-TOF MS tem sido considerada uma excelente opção para a caracterização bacteriana, por ser eficiente, rápida, de fácil aplicação e este estudo confirmou a sua elevada especificidade e sensibilidade na identificação de bactérias marinhas.(AU)

Microencapsulation of essential thyme oil by spray drying and its antimicrobial evaluation against Vibrio alginolyticus and Vibrio parahaemolyticus / Microencapsulação do óleo essencial de tomilho e avaliação de sua atividade antibacteriana sobre V. alginolyticus e V. parhaemolyticus

Abstract The aims of this research were first, to evaluate the antibacterial potential of commercial thyme essential oil against V. alginolyticus and V. parahaemolyticys and second, using the spray drying technique to produce microcapsules. chemical compounds of thyme oil and microcapsules were identified and quantified being thymol the chemical component present at the highest concentration. Oil-in-water (O/W) emulsions were prepared and the microcapsules were obtained with a spray dryer using maltodextrin as wall material (ratio 1:4). Thyme oil and the microcapsules exhibited antimicrobial activity against V. parahaemolyticus and V. alginolyticus. The spray drying process did not affect the antimicrobial activity of thyme essentialoil.

Resumo Os objetivos desta pesquisa foram avaliar o potencial antibacteriano do óleoessencial de tomilho sobre V. alginolyticus e V. parahaemolyticys e produzir microcápsulas através do processo de secagem por aspersão (spray dryer). Os compostos químicos do óleo essencial de tomilho e das microcápsulas foram identificados e quantificadaos. Foi preparada uma emulsão de óleo em água (O/A) e em seguida foram produzidas microcápsulas em um spray dryer utilizando-se óleo essencial de tomilho e maltodextrina como material de parede na proporção de 1:4 respectivamente. Entre os vários compostos identificados, o timol apresentou maior concentração. O óleo essencial de tomilho e as microcápsulas exibiram atividade antibacteriana sobre V. parahaemolyticus e V. alginolyticus. O processo de secagem por aspersão não afetou a atividade antibacteriana do óleo essencial de tomilho.

Molecular variations in Vibrio alginolyticus and V. harveyi in shrimp-farming systems upon stress

A study was performed to investigate the genomic variations in the shrimp farm isolates of Vibrio alginolyticus and V. harveyi when the isolates were subjected to environmental stress. Samples of shrimps, water and sediment were collected from Southern Indian coastal shrimp farms. Vibrio isolates were biochemically identified and confirmed using 16S rDNA and gyrB gene specific PCR. The bacterial strains were genotyped by PCR fingerprinting using GTG(5) and IS (Insertion Sequence) primers. Seven strains each of V. alginolyticus and V. harveyi were subjected to 10 passages through trypticase soya broth (TSB), which contained different NaCl concentrations (3, 6 and 8%) and trypticase soya agar (TSA). V. alginolyticus was also passaged through TSB with a 12% NaCl concentration. PCR fingerprinting, which was performed on the strains that were passaged through different salt concentrations, confirmed that V. alginolyticus and V. harveyi could affect the genomic variations, depending on the environmental conditions of the culture. The study highlights the complex genotypic variations that occur in Vibrio strains of tropical aquatic environment because of varied environmental conditions, which result in genetic divergence and/or probable convergence. Such genetic divergence and/or convergence can lead to the organismal adaptive variation, which results in their ability to cause a productive infection in aquatic organisms or generation of new strains.

Distribution of Vibrio alginolyticus-like species in Shenzhen coastal waters, China

We investigated the distribution of vibrios in Shenzhen coastal waters in order to obtain valuable information for the aquaculture industry and a health warning system. Quantities of vibrios from surface waters ranged from 0 to 4.40×10(4) CFUs mL-1 in April (spring), while from 0 to 2.57×10³ CFUs mL-1 in September (autumn); the abundance of V. alginolyticus-like species from surface water ranged from 0 to 6.72×10³ CFUs mL-1 in April (spring) and from 0 to 1.28×10³ CFUs mL-1 in September (autumn); higher counts were observed in spring. The V. alginolyticus-like species was dominant in Shenzhen coastal waters, with the highest abundance in the clean region (stations YMK001 and GDN064) in April, suggesting that Vibrio spp. were naturally occurring bacteria in marine environments. The correlation between the abundance of vibrios (including V. alginolyticus-like species) and environmental factors varied in different regions and different seasons. There were no vibrios detected when the salinity was less than 11.15ë in the Zhujiang River estuary, which indicated that salinity played a key role in the distribution of vibrios and V. alginolyticus-like species.

Isolation and experimental infection with Vibrio alginolyticus in the sea horse, Hippocampus reidi Ginsburg, 1933 (Osteichthyes: Syngnathidae) in Brazil / Isolamento e infecção experimental de Vibrio alginolyticus em cavalo-marinho, Hippocampus reidi Ginsburg, 1933 (Osteichthyes: Syngnathidae) no Brasil

The aim of this study was to evaluate the pathogenicity of Vibrio alginolyticus isolated from an outbreak of sea horse Hippocampus reidi reared in the State of Santa Catarina, Brazil, by experimental infection. Sea horses with necrosis on the mouth epithelium were collected from aquaria at the Aquaculture Department, UFSC and the bacterium isolated from the mouth, liver, heart and blood in tiosulphate citrate bilesalt sucrose agar broth. The strains were identified by API 20E kit with 99.1 percent probability as Vibrio alginolyticus. Twelve adult sea horses (9.63 ± 2.42 g and 15.12 ± 0.87 cm) were distributed in six aquaria of 10 L capacity with aerated sea water. Fish from three aquaria were submitted to an immersion bath in a solution containing 1.0 × 10(7) CFU of V. alginolyticus/mL for 15 minutes. Fish from the other three aquaria received the same procedure without bacteria. Twenty four hours after this challenge, 100 percent mortality was observed in the animals infected with V. alginolyticus. No mortality was observed in non-infected fish. Hyperplasia, displacement and fusion of secondary lamellae of the gills; leukocyte infiltration and necrotic foci in the kidney; hyperplasia, sinusoidal deformation and necrotic foci in the liver were observed in histopathological analysis. The V. alginolyticus isolated in this study was pathogenic to H. reidi and constitutes an important sanitary problem to its culture.

Foi avaliado por meio de infecção experimental a patogenicidade de Vibrio alginolyticus isolado de um surto de enfermidade em cavalo-marinho Hippocampus reidi cultivado no Estado de Santa Catarina, Brasil. Os peixes com necroses no epitélio bucal foram coletados em aquários do Departamento de Aquicultura, UFSC e as bactérias isoladas da boca, fígado, coração e sangue em meio Agar tiossulfato citrato bile sacarose. Os isolados foram identificados pelo kit API 20E como Vibrio alginolyticus com 99,1 por cento de probabilidade. Doze peixes adultos (9,63 ± 2,42 g e 15,12 ± 0,87 cm) foram distribuídos em seis aquários de 10 L com água marinha e aeração. Peixes de três aquários foram submetidos a um banho de imersão por 15 minutos em uma solução contendo 1,0 × 10(7) UFC de V. alginolyticus/mL. Nos outros três aquários realizou-se o mesmo procedimento sem a bactéria. Vinte e quatro horas após o desafio, 100 por cento de mortalidade foi observada nos animais infectados com V. alginolyticus. Não houve mortalidade nos peixes não infectados. Nas análises histopatológicas, foi observado hiperplasia, deslocamento do epitélio e fusão das lamelas secundárias das brânquias; infiltração de leucócitos e necrose no rim; hiperplasia, deformação sinusoidal e necrose no fígado nos animais desafiados com V. alginolyticus. O V. alginolyticus isolado neste estudo foi patogênico para H. reidi, constituindo-se de um importante problema para seu cultivo.

Adherence assays and Slime production of Vibrio alginolyticus and Vibrio parahaemolyticus / Ensaios de adesão e produção de muco por Vibrio alginolyticus e Vibrio parahaemolyticus

In this study we investigated the phenotypic slime production of Vibrio alginolyticus and Vibrioparahaemolyticus strains, food-borne pathogens, using a Congo red agar plate assay. Furthermore, westudied their ability to adhere to abiotic surfaces and Vero cells line. Our results showed that only V.alginolyticus ATCC 17749 was a slime-producer developing almost black colonies on Congo red agar plate.Adherence to glace tube showed that all V. alginolyticus strains were more adherent than V. parahaemolyticus.Only V. alginolyticus ATCC 17749 was found to be able to form biofilm on polystyrene microplate wells (OD570= 0.532). Adherence to Vero cells showed that all tested strains were non adherent after 30 min, however after60 min all the studied strains become adherent. The percentage of adherence ranged from1.23% to 4.66%.

Neste estudo, investigou-se a produção de muco por cepas de Vibrio alginolyticus e Vibrio parahaemolyticus através do teste em placa de ágar com vermelho congo. Estudou-se também a capacidade de adesão à superfícies abióticas e células Vero. Os resultados indicaram que somente V. alginolyticus ATCC 17749 produziu muco, formando colônias quase negras nas placas de ágar com vermelho congo. O teste de adesão a tubos de vidro indicou que as cepas de V. alginolyticus foram maisaderentes do que as de V. parahaemolyticus. Somente V. alginolyticus ATCC 17749 foi capaz de formar biofilme nos poços das microplacas de poliestireno (OD570=0,532). Testes de adesão a células Vero mostraram que nenhuma das cepas apresentou adesão em 30 min, mas todas aderiram após 60 min. Aporcentagem de adesão variou de 1,23% a 4,66%.

Enzymatic characterization of Vibrio alginolyticus strains isolated from bivalves harvested at Venice Lagoon (Italy) and Guanabara Bay (Brazil) / Caracterização enzimática de cepas de Vibrio alginolyticus isoladas de bivalves coletados na Lagoa de Veneza (Itália) e Baía de Guanabara (Brasil)

The aquatic ecosystem is the natural habitat of microorganisms including Vibrio and Aeromonas genus which are pathogenic to human and animals. In the present investigation the frequency of these bacteria and the enzymatic characteristics of 34 Vibrio alginolyticus strains isolated from bivalves harvested in Venice Lagoon (Italy) and Guanabara Bay (Brazil) were carried out from November 2003 to February 2004. The mussels' samples were submitted to enrichment in Alkaline Peptone Water (APW) added with 1 percent of sodium chloride (NaCl) and APW plus 3 percent NaCl incubated at 37 ºC for 18-24h. Following the samples were streaked onto TCBS Agar (Thiossulfate Citrate Bile Sucrose Agar) and the suspected colonies were submitted to biochemical characterization. Also, the Vibrio alginolyticus strains were evaluated to collagenase, elastase and chondroitinase production. The results showed the isolation of 127 microorganisms distributed as follows: 105 Vibrio strains such as V. alginolyticus (32.4 percent), V. harveyi (19 percent) and V. parahaemolyticus (7.6 percent), 20 Aeromonas strains and two Plesiomonas shigelloides were the main pathogens isolated. We observed the production of the three enzymes from V. alginolyticus strains considered as the main virulence factors of the bacteria, especially in cases of human dermatological infection.

O ecossistema aquático é o habitat natural de microrganismos incluindo aqueles dos gêneros Vibrio e Aeromonas os quais são patogênicos para o homem e animais. Na presente investigação foi avaliada a freqüência destas bactérias e a característica enzimática de 34 cepas de Vibrio alginolyticus isoladas de bivalves coletados na Lagoa de Venice (Itália) e Baía de Guanabara (Brasil) durante o período de Novembro-2003 a Fevereiro-2004. As amostras de mexilhões foram submetidas a enriquecimento em Água Peptonada Alcalina (APA) adicionada de 1 por cento de Cloreto de Sódio (NaCl) e APA com 3 por cento de NaCl (37 ºC/18-24h). Em seguida as amostras foram semeadas em Agar TCBS (Agar Tiossulfato Citrato Bile Sacarose) e as colônias suspeitas foram submetidas à caracterização bioquímica. As cepas de Vibrio alginolyticus foram avaliadas quanto à produção das enzimas colagenase, elastase e condroitinase. Os resultados demonstraram o isolamento de 127 microrganismos assim distribuídos: 105 cepas de Vibrio das quais V. alginolyticus (32,4 por cento), V. harveyi (19 por cento) e V. parahaemolyticus (7,6 por cento), 20 cepas de Aeromonas e 2 Plesiomonas shigelloides foram os principais patógenos isolados. Observou-se a produção das três enzimas a partir de V. alginolyticus, consideradas principais fatores de virulência da bactéria, em especial em casos de infecção dermatológica humana.

Septic Shock due to Vibrio alginolyticus in a Cirrhotic Patient: The First Case in Korea

We describe a case of septic shock due to Vibrio alginolyticus presenting with fever and bilateral leg pain. Despite intensive management with antibiotics and inotropic agents, the patient died from septic shock 1 day after hospitalization. V. alginolyticus was isolated from both leg wounds and a blood culture. To the best of our knowledge, this is the first reported case of V. alginolyticus bacteremia in Korea.

Isolamento de Vibrio spp. em mexilhões (Perna perna) coletados na região de Ponta de Itaipu, Niterói, RJ / Isolation of Vibrio spp. in bivalvia (Perna perna) collected in the region of Ponta de Itaipu, Niterói, RJ

O ecossistema marinho representa o habitat natural de diversas bactérias do gênero Vibrio, dentre as quais algumas com relevante potencial patogênico para o homem e animais. Devido ao crescente consumo de moluscos bivalves nas cidades litorâneas, foram avaliadas 15 amostras de mexilhões (Perna perna) coletados na região de Itaipu, Niterói, RJ, a fim de investigar a presença de Vibrio spp. As amostras de mexilhões foram submetidas a enriquecimento em Água Peptonada Alcalina (APA) adicionada de 1 por cento de NaCl (Cloreto de Sódio) e APA adicionada de 3 por cento NaCl, incubadas a 37°C por 24 horas. Em seguida, estes foram semeados em Agar Tiossulfato Citrato Bile Sacarose (TCBS) e as colônias suspeitas submetidas à caracterização bioquímica. Os resultados obtidos na presente investigação apontaram a presença das espécies V. harveyi, V. alginolyticus, V. carchariae e V. parahaemolyticus, cuja importância associa-se ao risco de infecções após o consumo de mexilhões sem cozimento ou parcialmente cozidos, o que aumenta sua relevância para a Saúde Pública, particularmente em indivíduos portadores de doenças crônico-degenerativas.

Vibrio spp. isolados de mamíferos marinhos capturados na região litorânea do Sudeste ao Sul do Brasil / Vibrio spp. isolated from marine mammals captured in coastal regions from southwestern to southern Brazil

Avaliou-se a incidência de Vibrio spp. a partir de lesões superficiais em mamíferos marinhos encalhados ou capturados em redes de pesca nas regiões litorâneas do Sudeste (Rio de Janeiro) e Sul (RS) do Brasil. Foram coletadas 198 amostras, pelas instituições de pesquisa DEENSP, GEMARS e Ceclimar, as quais foram enviadas ao Labent/IOC/FIOCruz, onde foram submetidas ao enriquecimento em Agua Peptonada Alcalina (APA) adicionada de 1 por cento e 3 por cento de NaCl e in-cubadas a 37°C por 18-24 horas. Em seqüência foram semeadas em meio Agar Tiossulfato Citrato Bile Sacarose (TCBS) e as colônias suspeitas submetidas à caracterização bioquímica. Foram isoladas 108 cepas bacterianas, destacando-se Vibrio alginolyticus, V. parahaemolyticus, V. vulnificus e V. fluvialis como os principais patógenos isolados. Os resultados obtidos apontam para a necessidade de implementar atividades de vigilância e monitorização bacteriológica, particularmente de espécies selvagens, e reforçar os programas de proteção ambiental em casos de mamíferos marinhos ameaçados de extinção.

In the present investigation was evaluated the incidence of Vibrio spp. from superficial lesions at marine mammals beached or captured by fishing net in the southwestern (RJ) and southern (RS) coastal regions of Brazil. One hundred and ninety eight swabs were collected by DEENSP, GEMARS and Ceclimar institutes and sent to Labent/IOC/FIOCruz where the samples were submitted to enrichment in Alkaline Peptone Water (APW) added with 1 percent and 3 percent of sodium chloride (NaCl) incubated at 37°C for 18-24 hours. After the samples were streaked onto Thiossulfate Citrate Bile Sucrose Agar (TCBS), the suspected colonies were submitted to biochemical characterization. The results showed 108 strains, and Vibrio alginolyticus, V. parahaemolyticus, V. vulnificus and V. fluvialis were the main pathogens isolated. These results appoint the importance of surveillance and microbiological monitoring accomplishment and reinforcement of environmental protective programs applied to marine mammals endangered with extinction.

Capacidad de bacterias halófilas para capturar sodio in vitro y su posible aplicación en bioremediación en suelos salinos-sódicos / Ability of halophile bacteria to capture in Vitro sodium and its possible application in bioremediation of saline-sodic soil

Las prácticas agrícolas inadecuadas han sido la causa del aumento de los suelos salinos–sódicos en todo el planeta. Su recuperación se ha hecho principalmente cambiando el sodio por otro catión, generalmente calcio, remediando en forma mecánica y usando plantas halotolerantes. Aunque estas prácticas controlan en alguna medida el problema, no se ha podido hacer una recuperación efectiva en estos suelos. Este trabajo presenta como alternativa, el uso de bioremediación con bacterias halófilas. El objetivo de la investigación fue evaluar la capacidad de cinco especies de bacterias halófilas para capturar iones sodio in vitro y diseñar una propuesta para su posible aplicación en bioremediación de suelos sódicos y salinos. La captura del sodio se demostró comparando la concentración inicial de la solución de sodio sin inocular y la concentración de la misma con la bacteria inoculada a través de la técnica de espectroscopia de absorción atómica. Las bacterias que demostraron la capacidad de captura de sodio in vitro fueron: Vibrio alginolyticus, Vibrio metschnikovii, Flavimonas oryzihabitans y Agrobacterium tumefasciens. Serratia marcescens no demostró captura. La comprobación de la captura de sodio permitió hacer dos propuestas: diseñar un bioreactor con un consorcio microbiano que incluya las bacterias que capturaron sodio en el experimento y bacterias autóctonas presentes en los suelos salinos y sódicos; utilizar ingeniería genética para implantar el gen de bomba de sodio de la especie con mejor captura, en especies nativas existentes en este tipo de suelos.

Misidentification as Vibrio alginolyticus with Vitek GNI+ Card in Three Cases / 대한임상미생물학회지

The introduction of a new, fully automated system into the clinical microbiology laboratory contributes to a rapid identification of microorganisms with accurate and reliable results, but such a system requires a high cost and additional tests for identification of some species. For instance, additional tests on oxidase, indole, motility, hemolysis, and pigmentation are needed in the correct identification by using Vitek GNI+ system (bioMerieux Vitek Inc., MO, USA). In particular, Vibrio and Aeromonas species are occasionally identified incorrectly when an automated system is used, and thus conventional biochemical tests may be more reliable in the identification of such species. We experienced three cases of incorrect identification of Vibrio parahaemolyticus, Vibrio cholerae, and Aeromonas veronii biovar sobria as Vibrio alginolyticus by using Vitek GNI+ card.

Estudo de Vibrio ssp. potencialmente patogênicos através de métodos moleculares / Study of virulence factors in Vibrio ssp through molecular methods

De forma similar ao V. cholerae, outros vibrios potencialmente patogênicos podem causar doença no homem em uma variedade de formas de quadros auto-limitantes até diarréia severa a cólera, septicemia, celulite e infecções de pele. Com o advento da biologia molecular, novas técnicas vêm sendo empregadas para o estudo do gênero Vibrio com o intuito de determinar presença de fatores de virulência, traçar rotas de transmissão, ou ainda como ferramenta no estudo epidemiológico destes organismos. O presente estudo visou pesquisar a presença de genes codificadores de virulência em cepas de V. cholerae provenientes de vários países, e ainda estudar do polimorfismo genético, com o emprego de iniciadores para seqüências ERIC e BOX e eletroforese em campo pulsado (PFGE), para determinar a relação entre cepas de V. cholerae, V. mimicus, V. parahemolyticus, V. fluviais e V. alginolyticus isolados de amostras de ostras e mexilhões no Brasil, e V. metschnikovii isolados de amostras de peixe isolados de outros países bem como com o Padrão ATCC de cada espécie. Foi realizada também, a confirmação da posição taxonômica de cepas de V. metschnikovii isolados de amostras de peixes, e a comparação dos padrões de bandas com isolados de outros países e cepa padrão da espécie. A posição taxonômica de cepas de V. metschnikovii foi confirmada através da Hibridização DNA/DNA, e o estudo do polimorfismo genético através de ERIC PCR, BOX PCR e eletroforese em campo pulsado demonstrou que essas são poderosas ferramentas para estudos epidemiológicos do gênero Vibrio.

A Case of Simultaneous Isolation of Vibrio parahaemolyticus and Vibrio alginolyticus / 대한임상미생물학회지

V.parahaemolyticus or V.alginolyticus infections are usually associated with consumption of raw or undercooked shellfish, contaminated food, and exposure of wounds to warm seawater. V.parahaemolyticus causes gastroenteritis(the most common syndrome), wound infections, and septicemia. V alginolyticus occasionally causes extraintestinal infections in humans. so far, the authors have not found the report of V.parahaemolyticus and V.alginolyticus isolation from a patient. So, we report a case of concurrent isolation of V.parahaemolyticus and V.alginolyticus from a patient who had a history of intestinal diarrhea and vomiting.

A Case of Simultaneous Isolation of Vibrio parahaemolyticus and Vibrio alginolyticus / 대한임상미생물학회지

V.parahaemolyticus or V.alginolyticus infections are usually associated with consumption of raw or undercooked shellfish, contaminated food, and exposure of wounds to warm seawater. V.parahaemolyticus causes gastroenteritis(the most common syndrome), wound infections, and septicemia. V alginolyticus occasionally causes extraintestinal infections in humans. so far, the authors have not found the report of V.parahaemolyticus and V.alginolyticus isolation from a patient. So, we report a case of concurrent isolation of V.parahaemolyticus and V.alginolyticus from a patient who had a history of intestinal diarrhea and vomiting.

Characterization of a Vibrio parahaemolyticus Phage Isolated from Marine

A novel bacteriophage, designated as VPP97, that infects the strains of Vibiro parahaemolyticus (hallophilic, Gram-negative bacterium) isolated most commonly from marine environments, has been discovered, and several of its properties have been determined. The plaques were clear and sized 0.6-1.0 mm in diameter. The virion forms a single band on 70% sucrose gradient and p1.50 CsC1 gradient by sucrose gradient centrifugation and CsCI gradient centrifugation respectively. It has a hexagonal head and a relatively long tail, as shown by electron microscopy. Vibrio alginolyticus, Vibrio fluvialis and Vibrio furnissii were also sensitive to this phage It was almost totally inactivated at 70 degree C and at pH below 5 or over 10. The nucleic acid of VPP97 is composed of DNA. The VPP97 had 9 specific structural proteins sized between 21.5 kDa and 97.4 kDa on SDS-PAGE. When V. parahaemolyticus cultures were treated with either phage VPP97 or one of the several antibiotics for 2 hours, the viable number of V. parahaemolyticus treated with the phage VPP97 is lower than that treated with chloramphenicol, erythromycin or penicillin, but not lower than that treated with tetracycline. Mice that have responded to the phage treatment revealed the lower numbers of V. parahaemolyticus in small intestine and less damage on small intestine compared to the untreated mice. Therefore, we suggest that the phage treatment appears effective to the infection by V. parahaemolyticus.

Characterization About Vibrio alginolyticus Phage Isolated from Marine Products

Two phages for the pathogenic V. alginolyticus were isolated from marine products. These 2 phages were examined temperature stability, pH stability, inactivation by UV irradiation, damage on restriction system of host cell, antibody production, structure protein analysis and western blotting assay. V. alginolyticus phages(VAPs) fomed plaques about 0.5 - 0.9mm in diameter and bands 50 - 60% in sucrose density gradient, VAPs were stable below 65'C, pH 5 - 10 and mostly inactivation by UV irradiation for 120sec. Latend period was 15 - 20 min. and burst size was 1.3 - 1.4 * 10 PFU/cell. Restriction system of V. alginolyticus isolated was mostly inactivated by 45C, 20min. heating. VAPs had 14 specific structural proteins and 5 proteins related to antibody production.

Electromicroscopic Characterization about Vibrio alginolyticus Phage Isolated from Marine Products

The study of bacteriophage began by F.W. Twort in 1915 and the lytic cycle recognized by d'Herellel in 1917. It repeated about the marine bacteriophage containing Vibrio phage by Smith, Spencer and Ju. Authors isolated 2 virulent phages for the pathogenic V. alginolyticus from marine products. These 2 phages were examined their ultrastructure & host-infection by elecron microscopy and in vivo test using skin of rats. V. alginolyticus phages(VAPs) fomed plaques about 0.5 - 0.9mm in diameter and bands 50 - 60% in sucrose density gradient. VAP had 50 - 120nm tail and 40 - 90nm head in diameter. In vivo test, using rat skin, as well as in vitro test VAP had the activity to V. alginolyticus isolated.

A Case of Vibrio alginolyticus Isolated from Otorrhea of Chronic Otitis Media / 감염

Vibrio alginolyticus is a microorganism of marine environment that occasionally occurs as a human pathogen. We isolated V. alginolyticus from a patient with otitis media. A 37-year-old man had been exposed to seawater one month before admission. The isolate showed typical biochemical and characteristics of this organism such as positive Voges-Proskauer reaction, fermentation of sucrose, growth on 10% sodium chloride media. In vitro susceptibility test shows the isolate was resistant to ampicillin and carbenicillin, but was susceptible to other antimicrobial agents. The patient improved with ciprofloxacin and ofloxacin therapy.

Enumeração e identificação de víbrios sacarose-positivos em amostras de lulas frescas comercializadas no município de Niterói-RJ / Enumeration and identification of sucrose-positive vibrios in samples of fresh squids commercialized at Niterói county-RJ

Foi efetuado um estudo sobre a enumeração e a identificação de víbrios sacarose-positivos em lulas frescas obtidas no comércio varejista do município de Niterói-RJ. No experimento, em 50 amostras de lulas, identificadas como pertencentes à espécie Oorytheutis brasiliensis Blainville, 1823, foram isoladas Unidades Formadoras de Colônias (UFC) sacarose-positivas em 28 (56%) amostras. Destas, foram identificados o Vibrio a/ginolyticus e víbrios do grupo NAG (não aglutináveis). A média dos Números Mais Prováveis (NMP) dos víbrios totais (com exceção da amostra com NMP > 2400 bacts./g) foi 101,70 bacts./g. Do total de 29 isolamentos nas 28 amostras, o V alginolyticus do grupo I de Heiberg obteve o maior percentual (62,07%), o V alginolyticus do grupo 111 apenas 10,30% e os víbrios NAG com 27,63%.

From 50 samples of squids of lhe species Dorytheutis brasiliensis Blainville, 1823, Colonies Formed Units (CFUs) sucrose-positives Víbrio alginolyticus and NAG vibrios (non-agglutinable) were isolated. The average of the Most Probabble Number of total vibrios (except the sample with MPN > 2400 bacts./g) was 101,70 bacts./g. From 28 samples, 29 isolations were carried out and lhe V. alginolyticus group I of Heiberg's Classification showed the greatest percentage (62,07%). The V. alginolyticus group 111 showed only 10,30% and NAG vibrios 27,63%.