Disseminated neoplasia (DN), a proliferative cell disorder of the circulatory system of bivalves, was first reported in oysters in 1969. Since that time, the disease has been determined to be transmissible through water-borne exposure, but the etiological agent has not been unequivocally identified. In order to determine if a viral agent, possibly a retrovirus, could be the causative agent of DN, transmission experiments were performed, using both a cell-free filtrate and a sucrose gradientpurified preparation of a cell-free filtrate of DN positive materials. Additionally, a PCR-enhanced reverse transcriptase assay was used to determine if reverse transcriptase was present in tissues or hemolyrnph from DN positive soft shell clams Mya arenana. DN was transmitted to healthy clams by injection with whole DN cells, but not with cell-free filtrates prepared from either tissues from DN positive clams, or DN cells. The cell-free preparations from DN-positive tissues and hemolymph having high levels of DN cells in circulation exhibited positive reactions in the PCR-enhanced reverse transcriptase assay. Cell-free preparations of hemolymph from clams having low levels of DN (<0.1% of cells abnormal), hemocytes from normal soft shell clams, and normal soft shell clam tissues did not produce a positive reaction in the PCR enhanced reverse transcriptase assay.
Linking marine epizootics to a specific aetiology is notoriously difficult. Recent diagnostic successes show that marine disease diagnosis requires both modern, cutting-edge technology (e.g. metagenomics, quantitative real-time PCR) and more classic methods (e.g. transect surveys, histopathology and cell culture). Here, we discuss how this combination of traditional and modern approaches is necessary for rapid and accurate identification of marine diseases, and emphasize how sole reliance on any one technology or technique may lead disease investigations astray. We present diagnostic approaches at different scales, from the macro (environment, community, population and organismal scales) to the micro (tissue, organ, cell and genomic scales). We use disease case studies from a broad range of taxa to illustrate diagnostic successes from combining traditional and modern diagnostic methods. Finally, we recognize the need for increased capacity of centralized databases, networks, data repositories and contingency plans for diagnosis and management of marine disease.
Worldwide transfer and introduction of non-indigenous species in ballast water causes significant environmental and economic impact. One way to address this problem is to remove or inactivate organisms that are found in ballast water. In this study, 3 experiments were conducted in Puget Sound, Washington, USA, using a prototype ozone treatment system installed on a commercial oil tanker, the S/T Tonsina. Treatment consisted of ozone gas diffused into a ballast tank for 5 and 10 h. Treatment and control tanks were sampled during the ozonation period for chemistry, culturable bacteria, phytoplankton and zooplankton. Selected fish and invertebrates were placed in cages deployed in the treatment and control tanks. Ozone introduced into seawater rapidly converts bromide (Br -) to bromines (HOBr/OBr -), compounds that are disinfectants. These were measured as total residual oxidant (TRO). Ozone treatment inactivated large portions of culturable bacteria, phytoplankton and zooplankton. The highest reductions observed were 99.99% for the culturable bacteria, > 99% for dinoflagellates and 96% for zooplankton. Caged animal results varied among taxa and locations in the ballast tank. Sheepshead minnows and mysid shrimp were most susceptible, shore crabs and amphipods the least. Distribution of ozone in the treatment tank was not homogenous during experiments, as suggested by the observed TRO concentrations and lower efficacies for inactivating the different taxa in selected ballast tank locations. Low concentrations of bromoform, a disinfection byproduct, were found in treated ballast water.
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