Production of positive controls for calcivirus-specific PCR using recombinant baculiovirus technology

Recent advances in our knowledge of the genetic structure of human caliciviruses (HuCVs) and small round-structured viruses (SRSVs) have led to the development of polymerase chain reaction (PCR)-based molecular tests specific for these viruses. These methods have been developed to detect a number of human pathogenic viruses in environmental samples including water, sewage and shellfish. HuCVs and SRSVs are not culturable, and no animal model is currently available. Therefore there is no convenient method of preparing viruses for study or for reagent production. One problem facing those attempting to use PCR-based methods for the detection of HuCVs and SRSVs is the lack of a suitable positive control substrate. This is particularly important when screening complex samples in which the levels of inhibitors present may significantly interfere with amplificiation. Regions within the RNA polymerase regions of two genetically distinct human caliciviruses have been amplified and used to produce recombinant baculoviruses which express RNA corresponding to the calicivirus polymerase. This RNA is being investigated as a positive control substrate for PCR testing, using current diagnostic primer sets. Recombinant baculovirus technology will enable efficient and cost-effective production of large quantities of positive control RNA with a specific known genotype. We consider the development of these systems as essential for successful screening and monitoring applications.

Human use pharmaceuticals in the estuarine environment: a survey of the Chesapeake Bay, Biscayne Bay, and Gulf of the Farallones

The assessment of emerging risks in the aquatic environment is a major concern and focus of environmental science (Daughton and Ternes, 1999). One significant class of chemicals that has received relatively little attention until recently are the human use pharmaceuticals. In 2004, an estimated 2.6 billion prescriptions were written for the top 300 pharmaceuticals in the U.S. (RxList, 2005). Mellon et al. (2001) estimated that 1.4 million kg of antimicrobials are used in human medicine every year. The use of pharmaceuticals is also estimated to be on par with agrochemicals (Daughton and Ternes, 1999). Unlike agrochemicals (e.g., pesticides) which tend to be delivered to the environment in seasonal pulses, pharmaceuticals are continuously released through the use/excretion and disposal of these chemicals, which may produce the same exposure potential as truly persistent pollutants. Human use pharmaceuticals can enter the aquatic environment through a number of pathways, although the main one is thought to be via ingestion and subsequent excretion by humans (Thomas and Hilton, 2004). Unused pharmaceuticals are typically flushed down the drain or wind up in landfills (Jones et al. 2001).

Cruise report: summer 2009 Sapelo Island National Estuarine Research Reserve ecological characterization (June 7 - June 13, 2009)

This cruise report is a summary of a field survey conducted within the Sapelo Island National Estuarine Research Reserve (SINERR), located on the Georgia coastline, June 7 – June 13, 2009. Multiple indicators of ecological condition and human dimensions were sampled synoptically at each of 30 stations throughout SINERR using a random probabilistic sampling design. Samples were collected for the analysis of benthic community structure and composition; concentrations of chemical contaminants (metals, pesticides, PAHs, PCBs, PBDEs) in sediments and target demersal biota; nutrient and chlorophyll levels in the water column; bacterial contaminants in the water column; and other basic habitat characteristics such as depth, salinity, temperature, dissolved oxygen, turbidity, total suspended solids, pH, sediment grain size, and organic carbon content. In addition to the fish samples that were collected for analysis of chemical contaminants relative to human-health consumption limits, other human-dimension indicators were sampled as well including presence or absence of fishing gear, vessels, surface trash, and noxious sediment odors. The overall purpose of the survey was to collect data to assess the status of ecosystem condition and potential stressor impacts throughout SINERR, based on these various indicators and corresponding management thresholds, and to provide this information as a baseline for determining how such conditions may be changing with time. While sample analysis is still ongoing a few preliminary results and observations are reported here. A final report will be completed once all data have been processed. The results will provide a comprehensive weight-of-evidence basis for evaluating current condition (aka a “state-of-the-SINEER environmental report”) and serve as a quantitative benchmark for tracking any future changes due to either natural or human disturbances. Another goal of the study is to demonstrate its utility as a possible model for assessing the status of condition at other NEERS sites using similar and consistent methods to promote system-wide regional and national comparisons.

The coastal environment and human health: microbial indicators, pathogens, sentinels and reservoirs

Innovative research relating oceans and human health is advancing our understanding of disease-causing organisms in coastal ecosystems. Novel techniques are elucidating the loading, transport and fate of pathogens in coastal ecosystems, and identifying sources of contamination. This research is facilitating improved risk assessments for seafood consumers and those who use the oceans for recreation. A number of challenges still remain and define future directions of research and public policy. Sample processing and molecular detection techniques need to be advanced to allow rapid and specific identification of microbes of public health concern from complex environmental samples. Water quality standards need to be updated to more accurately reflect health risks and to provide managers with improved tools for decision-making. Greater discrimination of virulent versus harmless microbes is needed to identify environmental reservoirs of pathogens and factors leading to human infections. Investigations must include examination of microbial community dynamics that may be important from a human health perspective. Further research is needed to evaluate the ecology of non-enteric water-transmitted diseases. Sentinels should also be established and monitored, providing early warning of dangers to ecosystem health. Taken together, this effort will provide more reliable information about public health risks associated with beaches and seafood consumption, and how human activities can affect their exposure to disease-causing organisms from the oceans.

Isolation of Salmonella braenderup (6, 7:e, h:e, n, z (sub15)) from Mussel meat

A brief account is given of the morphology, biochemistry and serology of Salmonella braenderup isolated from mussel meat, obtained from fish markets in Calicut, India. Possible implications regarding human salmonellosis are also considered.