Lethal marine snow : pathogen of bivalve mollusc concealed in marine aggregates

Author Posting. © The Authors, 2005. This is the author's version of the work. It is posted here by permission of American Society of Limnology and Oceanography for personal use, not for redistribution. The definitive version was published in Limnology and Oceanography 50 (2005): 1983-1988, doi:10.4319/lo.2005.50.6.1983.We evaluated marine aggregates as environmental reservoirs for a thraustochytrid pathogen, Quahog Parasite Unknown (QPX), of the northern quahog or hard clam, Mercenaria mercenaria. Positive results from in situ hybridization and denaturing gradient gel electrophoresis confirm the presence of QPX in marine aggregates collected from coastal embayments in Cape Cod, Massachusetts, where QPX outbreaks have occurred. In laboratory experiments, aggregates were observed and recorded by entering a quahog’s pallial cavity, thereby delivering embedded particles from the water column to its benthic bivalve host. The occurrence of pathogen-laden aggregates in coastal areas experiencing repeated disease outbreaks suggests a means for the spread and survival of pathogens between epidemics and provides a specific target for environmental monitoring of those pathogens.This work was funded by an NSF grant as part of the joint NSF-NIH Ecology of Infectious Disease program, by the Woods Hole Oceanographic Institution (WHOI) Sea Grant Program, under a grant from the National Oceanic and Atmospheric Administration, U.S. Department of Commerce, and a National Science Foundation Graduate Fellowship to M. Lyons

Bacterial Colonization and Extinction on Marine Aggregates: Stochastic Model of Species Presence and Abundance

Organic aggregates provide a favorable habitat for aquatic microbes, are efficiently filtered by shellfish, and may play a major role in the dynamics of aquatic pathogens. Quantifying this role requires understanding how pathogen abundance in the water and aggregate size interact to determine the presence and abundance of pathogen cells on individual aggregates. We build upon current understanding of the dynamics of bacteria and bacterial grazers on aggregates to develop a model for the dynamics of a bacterial pathogen species. The model accounts for the importance of stochasticity and the balance between colonization and extinction. Simulation results suggest that while colonization increases linearly with background density and aggregate size, extinction rates are expected to be nonlinear on small aggregates in a low background density of the pathogen. Under these conditions, we predict lower probabilities of pathogen presence and reduced abundance on aggregates compared with predictions based solely on colonization. These results suggest that the importance of aggregates to the dynamics of aquatic bacterial pathogens may be dependent on the interaction between aggregate size and background pathogen density, and that these interactions are strongly influenced by ecological interactions and pathogen traits. The model provides testable predictions and can be a useful tool for exploring how species-specific differences in pathogen traits may alter the effect of aggregates on disease transmission

Bacterial Colonization and Extinction on Marine Aggregates: Stochastic Model of Species Presence and Abundance

Organic aggregates provide a favorable habitat for aquatic microbes, are efficiently filtered by shellfish, and may play a major role in the dynamics of aquatic pathogens. Quantifying this role requires understanding how pathogen abundance in the water and aggregate size interact to determine the presence and abundance of pathogen cells on individual aggregates. We build upon current understanding of the dynamics of bacteria and bacterial grazers on aggregates to develop a model for the dynamics of a bacterial pathogen species. The model accounts for the importance of stochasticity and the balance between colonization and extinction. Simulation results suggest that while colonization increases linearly with background density and aggregate size, extinction rates are expected to be nonlinear on small aggregates in a low background density of the pathogen. Under these conditions, we predict lower probabilities of pathogen presence and reduced abundance on aggregates compared with predictions based solely on colonization. These results suggest that the importance of aggregates to the dynamics of aquatic bacterial pathogens may be dependent on the interaction between aggregate size and background pathogen density, and that these interactions are strongly influenced by ecological interactions and pathogen traits. The model provides testable predictions and can be a useful tool for exploring how species-specific differences in pathogen traits may alter the effect of aggregates on disease transmission

Development of a real time quantitative PCR assay for the hard clam pathogen Quahog Parasite Unknown (QPX)

Author Posting. © Inter-Research, 2006. This article is posted here by permission of Inter-Research for personal use, not for redistribution. The definitive version was published in Diseases of Aquatic Organisms 72 (2006): 45-52, doi:10.3354/dao072045.Quahog Parasite Unknown (QPX) is a thraustochytrid pathogen responsible for catastrophic mortalities of the northern quahog (hard clam) Mercenaria mercenaria. A real-time quantitative polymerase chain reaction (qPCR) assay was developed to assist research efforts on QPX ecology and pathology. Sensitivity of the assay was evaluated with serial dilutions of QPX-cultured cells to determine the lowest concentration of DNA that remained detectable in both the presence and absence of extraneous environmental substances. QPX cells were quantified before DNA extraction to calibrate standard curves to cell counts. Based on our results, the qPCR assay is able to quantify QPX within the range of 1 to several thousand organisms per reaction. Specificity of the assay was assessed by testing 29 thraustochytrid-like protists isolated from suspension-feeding bivalves from China, Oregon, Maryland, and Virginia. Application of the assay was demonstrated with positive qPCR results from naturally contaminated environmental samples including marine aggregates (i.e. marine snow), clam pseudofeces, and inflammatory nodules from infected clams. This quantitative assay for QPX will provide a valuable tool for characterizing QPX parasite abundances in coastal environments and for improving clam disease diagnostics.This research was funded in part by NSF-NIH Ecology of Infectious Disease Grant (No. 0429018) to R.S. and a grant to R.S. and S.B.R. from the County of Barnstable, Massachusetts

Environmental distribution and persistence of Quahog Parasite Unknown (QPX)

Author Posting. © Inter-Research, 2008. This article is posted here by permission of Inter-Research for personal use, not for redistribution. The definitive version was published in Diseases of Aquatic Organisms 81 (2008): 219-229, doi:10.3354/dao01948.Quahog Parasite Unknown (QPX) is the cause of mass mortality events of hard clams Mercenaria mercenaria from Virginia, USA, to New Brunswick, Canada. Aquaculture areas in Massachusetts, USA, have been particularly hard hit. The parasite has been shown to be a directly infective organism, but it is unclear whether it could exist or persist outside of its clam host. We used molecular methods to examine water, sediment, seaweeds, seagrass and various invertebrates for the presence of QPX. Sites in Virginia and Massachusetts were selected based upon the incidence of QPX-induced clam die-offs, and they were monitored seasonally. QPX was detectable in almost all of our different sample types from Massachusetts, indicating that the parasite was widely distributed in the environment. Significantly more samples from Massachusetts were positive than from Virginia, and there was a seasonal pattern to the types of samples positive from Massachusetts. The data suggest that, although it may be difficult to completely eradicate QPX from the environment, it may be possible to keep the incidence of disease under control through good plot husbandry and the removal of infected and dying clams.This work is the result of research sponsored by NOAA National Sea Grant College Program Office, Department of Commerce, under Grant No. NA16RG2273, Woods Hole Oceanographic Institution Sea Grant Project No. R/B-168

Quantification of recA gene expression as an indicator of repair potential in marine bacterioplankton communities of Antarctica

Marine bacteria in surface waters must cope daily with the damaging effects of exposure to solar radiation (containing both UV-A and UV-B wavelengths), which produces lesions in their DNA. As the stratospheric ozone layer is depleted, these coping mechanisms are likely to play an even more important role in the viability of marine bacterial communities. The recA gene is ubiquitous among eubacteria and is highly conserved both in nucleotide and amino acid sequence. Besides its role in generalized recombination, the gene's translational product, RecA, is the regulator of 'dark repair' activity (DNA-repair mechanisms that do not require visible light as a cofactor). We have taken advantage of this function and used recA gene expression as a barometer of the DNAdamage repair capacity of bacterial assemblages in the Southern Ocean. Studies were conducted in the Gerlache Strait, Antarctica, in the austral springs of 1995 and 1996. Analysis of both recA mRNA and RecA protein extracted from natural communities indicated that the level of expression of this gene varied in a diel fashion, suggesting an increased repair capacity in these organisms. These included an early morning rise in RecA levels followed by a plateau or even a reduction in RecA concentration during the remainder of the day. A much greater increase in RecA was consistently observed after sunset, followed by a constant decrease during the night. Microcosm experiments with a RecA+ Gerlache Strait g-proteobacteria isolate, RM11001, demonstrated a similar diel pattern of expression. These studies demonstrate the usefulness of RecA as a biological indicator of DNA repair capacity in natural bacterial assemblages. They indicate that 'dark repair' of DNA damage is an important coping mechanism for bacteria in the marine environment of Antarctica.Peer reviewedMicrobiology and Molecular Genetic