Characterization of microbial communities from coastal waters using microarrays

Molecular methods, including DNA probes, were used to identify and enumerate pathogenic Vibrio species in the Chesapeake Bay; our data indicated that Vibrio vulnificus exhibits seasonal fluctuations in number. Our work included a characterization of total microbial communities from the Bay; development of microarrays that identify and quantify the diversity of those communities; and observation of temporal changes in those communities. To identify members of the microbial community, we amplified the 16S rDNA gene from community DNA isolated from a biofilm sample collected from the Chesapeake Bay in February, 2000. The resultant 75 sequences were 95% or more similar to 7 species including two recently described Shewanella species, baltica and frigidimarina, that have not been previously isolated from the Chesapeake. When the genera of bacteria from biofilm after culturing are compared to those detected by subcloning amplified 16S fragments from community DNA, the cultured sample exhibited a strong bias. In oysters collected in February, the most common bacteria were previously unknown. Based on our 16S findings, we are developing microarrays to detect these and other microbial species in these estuarine communities. The microarrays will detect each species using four distinct loci, with the multiple loci serving as an internal control. The accuracy of the microarray will be measured using sentinel species such as Aeromonas species, Escherichia coli, and Vibrio vulnificus. Using microarrays, it should be possible to determine the annual fluctuations of bacterial species (culturable and non-culturable, pathogenic and non-pathogenic). The data may be applied to understanding patterns of environmental change; assessing the health of the Bay; and evaluating the risk of human illness associated with exposure to and ingestion of water and shellfish.     

Ultrafine particle fluxes above four major European cities

Little information is known about the behaviour of ultrafine particles (UFP) on a citywide scale. Total particle number flux, dominated by UFP, and other meteorological parameters were collected from tower sites in Manchester, London, Edinburgh and Gothenburg between 1999 and 2006 using the eddy covariance technique. Averaged diurnal cycles were produced for particle number flux, concentration, sensible heat flux, emission velocity, friction velocity, wind speed and temperature. UFP flux cycles showed clear diurnal trends which were linked to traffic activity and local sources. Wind sector analysis showed contributions to flux from local heavily urbanised areas. A simple parameterised model linking UFP flux to traffic activity, sensible heat and friction velocity above the city was produced.     

An assessment of fecal indicator and other bacteria from an urbanized coastal lagoon in the City of Los Angeles, California, USA

A study was performed in Del Rey Lagoon, City of Los Angeles, to determine if the lagoon was as a source or sink for fecal indicator bacteria (FIB: total coliforms, Escherichia coli, enterococci) and to screen for the presence of other potentially pathogenic bacteria. The lagoon receives tidal flows from the adjacent Ballona Estuary whose water usually is contaminated with FIB originating from the highly urbanized Ballona Creek Watershed. During 16 sampling events from February 2008 through March 2009, replicate water samples (n?=?3) were collected 1 h prior to the high tide and 1 h prior to the following low tide. FIB concentrations were measured by the defined substrate method (IDEXX, Westbrook, Me) followed by culturing of bacterial isolates sampled from positive IDEXX Quanti-Tray wells and were identified using the Vitek 2 Compact (bioMérieux, Durham, NC). Mean concentrations of FIB often differed by an order of magnitude from flood to ebb flow conditions. The lagoon tended to act as a sink for total coliforms based on the ratio of mean flood to ebb densities (R _F/E) >1.0 during 56 % of the sampling events and during ebb flows, as a source for E. coli and enterococci (R _F/E <1.69 % of events). Approximately 54 species were identified from 277 isolates cultured from the IDEXX Quanti-Trays. Of these, 54 % were species known to include pathogenic strains that can be naturally occurring, introduced in runoff, or originated from other sources. Diversity and cluster analyses indicated a dynamic assemblage that changes in species composition with day-to-day fluctuations as well as tidal action. The concept of monitoring the lagoon and estuary as a sentinel habitat for pathogenic assemblages is discussed.     

Microbiological Monitoring of Marine Recreational Waters in Southern California

An inventory was conducted to assess the number, type, spatial distribution, and costs of microbiological monitoring programs in southern California marine waters from Point Conception to the US/Mexico International Border. The location of each sampling site was determined using global positioning system (GPS), and estimates of geographic coverage were determined using geographic information system (GIS) techniques. Twenty-one programs conducted 87,007 tests annually at 576 sites in the study area. The largest number of sites was sampled in Orange County, whereas the largest number of analyses was performed in Los Angeles County because monitoring programs in this area focused on daily monitoring. Fifteen of the 21 programs were managed by National Pollutant Discharge Elimination System (NPDES) permitted sewage effluent dischargers who sampled both offshore and shoreline waters and typically tested for three indicator bacteria (total coliform, fecal coliform, and enterococcus). Their combined efforts comprised 82% of all of the microbiological indicator analyses conducted on an annual basis. Five of the remaining monitoring organizations were public health agencies, which typically focus their efforts on testing only total coliforms. Laboratory methodology also varied considerably, with NPDES permittees predominantly utilizing membrane filtration while public health agencies generally used multiple tube fermentation or premanufactured test kits. Nearly three quarters of all the effort expended in southern California occurred along the shoreline as opposed to offshore locations. Two thirds of this shoreline effort was focused on high-use sandy beaches and in proximity to perennial freshwater outlets (storm drains and creeks). Most sampling occurred at a set of fixed sites that were revisited frequently, but only represented about 7% of the total shoreline. We estimated that roughly $3 million is spent annually on monitoring bathing water quality in southern California, exceeding that spent in any other part of the country.     

Improving water quality through California’s Clean Beach Initiative: an assessment of 17 projects

California’s Clean Beach Initiative (CBI) funds projects to reduce loads of fecal indicator bacteria (FIB) impacting beaches, thus providing an opportunity to judge the effectiveness of various CBI water pollution control strategies. Seventeen initial projects were selected for assessment to determine their effectiveness on reducing FIB in the receiving waters along beaches nearest to the projects. Control strategies included low-flow diversions, sterilization facilities, sewer improvements, pier best management practices (BMPs), vegetative swales, and enclosed beach BMPs. Assessments were based on statistical changes in pre- and postproject mean densities of FIB at shoreline monitoring stations targeted by the projects. Most low-flow diversions and the wetland swale project were effective in removing all contaminated runoff from beaches. UV sterilization was effective when coupled with pretreatment filtration and where effluent was released within a few hundred meters of the beach to avoid FIB regrowth. Other BMPs were less effective because they treated only a portion of contaminant sources impacting their target beach. These findings should be useful to other coastal states and agencies faced with similar pollution control problems.     

A Regional Survey of the Microbiological Water Quality along the Shoreline of the Southern California Bight

A regional survey of the microbiological water quality along the shoreline of the Southern California Bight (SCB), from Point Conception south to Ensenada, Mexico, was conducted during August, 1998, by 36 agencies under the coordination of the Southern California Coastal Water Research Project (SCCWRP). Microbiological water quality was assessed by calculating the percentage of shoreline-mile-days that exceeded bacterial indicator thresholds for total and fecal coliforms, total/fecal ratios, and enterococci. Sample sites were selected using a stratified random sampling approach, with the SCB recreational shoreline divided into six strata: high- and low-use sandy beaches, high- and low-use rocky shoreline, and perennial and ephemeral freshwater outlets. Samples were collected on a weekly basis at a total of 253 sites, beginning on August 2^nd, 1998 and continuing for five weeks. Samples were analyzed by 22 participating labs using their normal methods (multiple tube fermentation, membrane filtration, Colilert® and/or Enterolert®). All labs met testing criteria established through intercalibration exercises and quality control check samples distributed during the sampling period. Nearly 95% of the shoreline-mile days did not exceed daily and monthly bacterial indicator thresholds, demonstrating good bacteriological water quality along the SCB shoreline. Freshwater outlets, comprised mainly of storm drains, had the poorest water quality with 60% and 40% of the shoreline-miles exceeding monthly and daily thresholds, respectively. Freshwater outlets were also more likely to demonstrate exceedances by multiple indicators at a single site, and repeat exceedances at sites over the five-week period. Compared with the southern California beaches, Mexican beaches had nearly 5 times the number of exceedances for total and fecal coliforms, and nearly 8 times the number of exceedances for total/fecal ratios.