Microbial community analysis of Acropora palamata mucus swabs, water and sediment samples from Hawksnest Bay, St. John, U.S. Virgin Islands

Colonies of the scleractinian coral Acropora palmata, listed as threatened under the US Endangered Species Act in 2006, have been monitored in Hawksnest Bay, within Virgin Islands National Park, St. John, from 2004 through 2010 by scientists with the US Geological Survey, National Park Service, and the University of the Virgin Islands. The focus has been on documenting the prevalence of disease, including white band, white pox (also called patchy necrosis and white patches), and unidentified diseases (Rogers et al., 2008; Muller et al., 2008). In an effort to learn more about the pathologies that might be involved with the diseases that were observed, samples were collected from apparently healthy and diseased colonies in July 2009 for analysis. Two different microbial assays were performed on Epicentre Biotechnologies DNA swabs containing A. palmata coral mucus, and on water and sediment samples collected in Hawksnest Bay. Both assays are based on polymerase chain reaction (PCR) amplification of portions of the small rRNA gene (16S). The objectives were to determine 1) if known coral bacterial pathogens Serratia marcescens (Acroporid Serratiosis), Vibrio coralliilyticus (temperature-dependent bleaching, White Syndrome), Vibrio shiloi (bleaching, necrosis), and Aurantimonas coralicida (White Plague Type II) were present in any samples, and 2) if there were any differences in microbial community profiles of each healthy, unaffected or diseased coral mucus swab. In addition to coral mucus, water and sediment samples were included to show ambient microbial populations. In the first test, PCR was used to separately amplify the unique and diagnostic region of the 16S rRNA gene for each of the coral pathogens being screened. Each pathogen test was designed so that an amplified DNA fragment could be seen only if the specific pathogen was present in a sample. A positive result was indicated by bands of DNA of the appropriate size on an agarose gel, which separates DNA fragments based on the size of the molecule. DNA from pure cultures of each of the pathogens was used as a positive control for each assay.

Viruses: agents of coral disease?

The potential role of viruses in coral disease has only recently begun to receive attention. Here we describe our attempts to determine whether viruses are present in thermally stressed corals Pavona danai, Acropora formosa and Stylophora pistillata and zoanthids Zoanthus sp., and their zooxanthellae. Heat-shocked P. danai, A. formosa and Zoanthus sp. all produced numerous virus-like particles (VLPs) that were evident in the animal tissue, zooxanthellae and the surrounding seawater; VLPs were also seen around heat-shocked freshly isolated zooxanthellae (FIZ) from P. danai and S. pistillata. The most commonly seen VLPs were tail-less, hexagonal and about 40 to 50 nm in diameter, though a diverse range of other VLP morphotypes (e.g. rounded, rod-shaped, droplet-shaped, filamentous) were also present around corals. When VLPs around heat-shocked FIZ from S. pistillata were added to non-stressed FIZ from this coral, they resulted in cell lysis, suggesting that an infectious agent was present; however, analysis with transmission electron microscopy provided no clear evidence of viral infection. The release of diverse VLPs was again apparent when flow cytometry was used to enumerate release by heat-stressed A. formosa nubbins. Our data support the infection of reef corals by viruses, though we cannot yet determine the precise origin (i.e. coral, zooxanthellae and/or surface microbes) of the VLPs seen. Furthermore, genome sequence data are required to establish the presence of viruses unequivocally.

The characteristics and questionable taxonomic position of the oxalate-decomposing bacterium, Vibrio extorquens

A detailed account of the morphological, staining, penicillin sensitivity and serological peculiarities of five strains of an oxalate-decomposing bacterium including the well-recognized strain Vibrio extorquens, has been given. Inasmuch as all the strains share many of the characteristics of the genus Arthrobacter and not Vibrio the desirability of placing the bacterium in the former genus for the time being has been suggested. The possibility of the strains falling under an altogether new genus which represent a phylogenic link between the pseudomonads and diphtheroids has been speculated.

Influencia de las bajas temperaturas en la supervivencia y el perfil proteico de Vibrio harveyi

Poster presentado al XXII Congreso Nacional de Microbiología celebrado en Salamanca los días 11-14 julio de 2011.

Liberación de compuestos al medio circundante durante la supervivencia a bajas temperaturas en Vibrio harveyi

Vibrio harveyi es considerado como una de las especies más relevantes del género Vibrio debido a su capacidad para infectar peces marinos e invertebrados. Estudios previos han demostrado que la respuesta de V. harveyi ante condiciones ambientales adversas (p.e. disminución de la temperatura) es su entrada en el denominado estado Viable No Cultivable (VNC), representando este estado una estrategia de supervivencia para algunas bacterias no diferenciadas. Se ha estudiado la respuesta de este microorganismo durante su incubación a bajas temperaturas (4˚C) utilizando como soporte tanto agua de mar como sobrenadantes recogidos en experiencias de superviviencia previas. V. harveyi presenta un patrón similar durante su incubación en agua de mar como en fases tempranas de estudio en sobrenadantes. Sin embargo, en fases tardías de estudio se ha comprobado que se retrasa su entrada en el estado VNC. Estos resultados sugieren que estas poblaciones podrían liberan compuestos al medio para favorecer su supervivencia bajo condiciones ambientales adversas.

Efecto de la radiación visible sobre la supervivencia de Vibrio harveyi en el medio marino

Vibrio harveyi es un microorganismo marino perteneciente a la familia Vibrionaceae, patógeno de numerosos animales marinos; tanto invertebrados como vertebrados, pudiendo producir pérdidas económicas en países que se benefician de la acuicultura. Se trata de un microorganismo que vive en un medio natural con escasa cantidad de nutrientes, por ello es un microorganismo oligotrofo. Además el medio marino es un medio con una gran cantidad de sales, con lo cual V. harveyi es una bacteria halófila. 3 V. harveyi es capaz de entrar en lo que se conoce como estado Viable No Cultivable (VNC), en dicho estado es capaz de sobrevivir a situaciones de estrés manteniendo niveles bajos de actividad y perdiendo la cultivabilidad. La radiación luminosa visible, a pesar de tener efectos beneficiosos en los seres vivos, puede provocar efectos negativos en las poblaciones microbianas marinas. En este trabajo se determinó la entrada en estado VNC en sus condiciones de temperatura ambiente (20ºC) tanto en un control en oscuridad así como bajo estrés lumínico. Los resultados mostraron como las células mantenidas en oscuridad no entraron en estado VNC, aunque sí se produjo una pérdida de cultivabilidad relacionada con lesiones celulares provocadas por los nutrientes de determinados medios de cultivo. En cambio, las células que fueron expuestas a la luz visible indicaron una pérdida de cultivabilidad a lo largo de los días de exposición, manteniéndose al finalizar el trabajo experimental el 93% de la población en estado VNC. Por lo tanto, la luz visible provoca un efecto negativo en la población de V. harveyi que es capaz de mantenerse en un estado VNC para sobrevivir a las condiciones adversas.

Avaliação de sistema de cultivo integrado, a partir da reciclagem de águas residuais submetidas a tratamento primário: pesquisa de espécies dos gêneros Salmonella, Shigella, Vibrio e Aeromonas

Para avaliar um sistema integrado de aquicultura foram realizadas análises microbiológicas da água utilizada neste sistema e determinada a incidência e resistência antimicrobiana dos enteropatógenos no ecossistema relacionado. As amostras de água testadas apresentaram 32,9% de taxas de coliformes fecais (≤1.600/100mL), de acordo com a OMS para piscicultura em águas residuais. Salmonella spp. foram detectadas em 14,5% das amostras. De um total de 33 cepas, 15,1% eram resistentes a um ou dois antimicrobianos testados e resistência a múltiplas drogas não foi observada. Aeromonas spp. foram identificadas em 91,6% das amostras. De um total de 416 cepas, resistência a uma classe de antimicrobianos foi observada em 66,3% e a multirresistência às drogas em 37,7%. Na avaliação da virulência dos isolados de Aeromonas hydrophila, 85,3% das cepas apresentaram Beta-hemólise nos três diferentes tipos de eritrócitos empregados e 99,1% nos eritrócitos de coelho e cavalo, sendo possível a caracterização através da PCR do gene aerA e lip, em 100% das amostras. Os resultados obtidos apontam para a relevância quanto às vantagens da implementação de um sistema integrado, disponibilizando alimentos com custo reduzido, porém este sistema necessita de um controle rígido e efetivo para que estes produtos não constituam veículos para a disseminação de doenças.

Variations temporelles de Vibrio cholerae Non 01 et conditions environnementales dans un site aquacole (Lagune Ebrié, Côte d'Ivoire)

The annual cycle of Vibrio cholerae in the environment surrounding the Layo aquaculture facility (Ebrié lagoon) was studied from March 1991 to April 1992. Vibrio cholerae counts were coupled with the determination of physical and chemical characteristics of water and the estimation of biological richness of this environment.

The Demand for Eastern Oysters, Crassostrea virginica, from the Gulf of Mexico in the Presence of Vibrio vulnificus

California, in response to health concerns, initiated a program on 1 March 1991 which required anyone selling eastern oysters, Crassostrea virginica, from the Gulf of Mexico area to notify potential consumers that there was a risk in consuming them raw. This mandatory warning, followed shortly thereafter by a similar warning in other states, including Louisiana and Florida, received extensive press cover-age throughout the country and particularly in the Gulf area. This paper examines the extent to which the demand for Gulf-area oysters has been reduced as a result of mandatory warning labels and negative publicity. In general, the results suggest that since 1991 the “summer” dockside price has been reduced by about 50% as a result of warning labels and associated negative publicity, while the “winter” dockside price has been reduced by about 30%.

Predicting the distribution of Vibrio vulnificus in Chesapeake Bay

Vibrio vulnificus is a gram-negative pathogenic bacterium endemic to coastal waters worldwide, and a leading cause of seafood related mortality. Because of human health concerns, understanding the ecology of the species and potentially predicting its distribution is of great importance. We evaluated and applied a previously published qPCR assay to water samples (n = 235) collected from the main-stem of the Chesapeake Bay (2007 – 2008) by Maryland and Virginia State water quality monitoring programs. Results confirmed strong relationships between the likelihood of Vibrio vulnificus presence and both temperature and salinity that were used to develop a logistic regression model. The habitat model demonstrated a high degree of concordance (93%), and robustness as subsequent bootstrapping (n=1000) did not change model output (P > 0.05). We forced this empirical habitat model with temperature and salinity predictions generated by a regional hydrodynamic modeling system to demonstrate its utility in future pathogen forecasting efforts in the Chesapeake Bay.