A review of technologies for rapid detection of bacteria in recreational waters

Monitoring of recreational beaches for fecal indicator bacteria is currently performed using culturebased technology that can require more than a day for laboratory analysis, during which time swimmers are at risk. Here we review new methods that have the potential to reduce the measurement period to less than an hour. These methods generally involve two steps. The first is target capture, in which the microbial group of interest (or some molecular/chemical/or biochemical signature of the group) is removed, tagged or amplified to differentiate it from the remaining material in the sample. We discuss three classes of capture methods: 1) Surface and whole-cell recognition methods, including immunoassay techniques and molecule-specific probes; 2) Nucleic acid methods, including polymerase chain reaction (PCR), quantitative PCR (Q-PCR), nucleic acid sequence based amplification (NASBA) and microarrays; and 3) Enzyme/substrate methods utilizing chromogenic or fluorogenic substrates. The second step is detection, in which optical, electrochemical or piezoelectric technologies are used to quantify the captured, tagged or amplified material. The biggest technological hurdle for all of these methods is sensitivity, as EPA's recommended bathing water standard is less than one cell per ml and most detection technologies measure sample volumes less than 1 ml. This challenge is being overcome through addition of preconcentration or enrichment steps, which have the potential to boost sensitivity without the need to develop new detector technology. The second hurdle is demonstrating a relationship to health risk, since most new methods are based on measuring cell structure without assessing viability and may not relate to current water quality standards that were developed in epidemiology studies using culture-based methods. Enzyme/substrate methods may be the first rapid methods adopted because they are based on the same capture technology as currentlyapproved EPA methods and their relationship to health risk can be established by demonstrating equivalency to existing procedures. Demonstration of equivalency may also be possible for some surface and whole-cell recognition methods that capture bacteria in a potentially viable state. Nucleic acid technologies are the most versatile, but measure nonviable structure and will require inclusion in epidemiological studies to link their measurement with health risk

Storm Effects on Regional Beach Water Quality Along the Southern California Shoreline

Two regional studies conducted during dry weather demonstrated that the Southern California Bight (SCB) shoreline has good water quality, except near areas that drain land-based runoff. Here, we repeat those regional studies 36 h after a rainstorm to assess the influence of runoff under high flow conditions. Two hundred and fifty-four shoreline sites between Santa Barbara, California and Ensenada, Mexico were sampled using a stratified-random sampling design with four strata: sandy beaches, rocky shoreline, shoreline adjacent to urban runoff outlets that flow intermittently, and shoreline adjacent to outlets that flow year-round. Each site was sampled for total coliforms, fecal coliforms (or E. coli), and enterococci. Sixty percent of the shoreline failed water quality standards after the storm compared to only 6% during dry weather. Failure of water quality standards increased to more than 90% for shoreline areas adjacent to urban runoff outlets. During dry weather, most water quality failures occurred for only one of the three bacterial indicators and concentrations were barely above State of California standards; following the storm, most failures were for multiple indicators and exceeded State of California standards by a large margin. The condition of the shoreline in Mexico and the United States was similar following rainfall, which was not the case during dry weather

Storm effects on regional beach water quality along the southern California shoreline

Two regional studies conducted during dry weather demonstrated that the Southern California Bight (SCB) shoreline has good water quality, except near areas that drain land-based runoff. Here, we repeat those regional studies 36 h after a rainstorm to assess the influence of runoff under high flow conditions. Two hundred and fifty-four shoreline sites between Santa Barbara, California and Ensenada, Mexico were sampled using a stratified-random sampling design with four strata: sandy beaches, rocky shoreline, shoreline adjacent to urban runoff outlets that flow intermittently, and shoreline adjacent to outlets that flow year-round. Each site was sampled for total coliforms, fecal coliforms (or E. coli), and enterococci. Sixty percent of the shoreline failed water quality standards after the storm compared to only 6% during dry weather. Failure of water quality standards increased to more than 90% for shoreline areas adjacent to urban runoff outlets. During dry weather, most water quality failures occurred for only one of the three bacterial indicators and concentrations were barely above State of California standards; following the storm, most failures were for multiple indicators and exceeded State of California standards by a large margin. The condition of the shoreline in Mexico and the United States was similar following rainfall, which was not the case during dry weather

Using rapid indicators for Enterococcus to assess the risk of illness after exposure to urban runoff contaminated marine water

Background: Traditional fecal indicator bacteria (FIB) measurement is too slow (>18 h) for timely swimmer warnings. Objectives: Assess relationship of rapid indicator methods (qPCR) to illness at a marine beach impacted by urban runoff. Methods: We measured baseline and two-week health in 9525 individuals visiting Doheny Beach 2007-08. Illness rates were compared (swimmers vs. non-swimmers). FIB measured by traditional (Enterococcus spp. by EPA Method 1600 or Enterolert™, fecal coliforms, total coliforms) and three rapid qPCR assays for Enterococcus spp. (Taqman, Scorpion-1, Scorpion-2) were compared to health. Primary bacterial source was a creek flowing untreated into ocean; the creek did not reach the ocean when a sand berm formed. This provided a natural experiment for examining FIB-health relationships under varying conditions. Results: We observed significant increases in diarrhea (OR 1.90, 95% CI 1.29-2.80 for swallowing water) and other outcomes in swimmers compared to non-swimmers. Exposure (body immersion, head immersion, swallowed water) was associated with increasing risk of gastrointestinal illness (GI). Daily GI incidence patterns were different: swimmers (2-day peak) and non-swimmers (no peak). With berm-open, we observed associations between GI and traditional and rapid methods for Enterococcus; fewer associations occurred when berm status was not considered. Conclusions: We found increased risk of GI at this urban runoff beach. When FIB source flowed freely (berm-open), several traditional and rapid indicators were related to illness. When FIB source was weak (berm-closed) fewer illness associations were seen. These different relationships under different conditions at a single beach demonstrate the difficulties using these indicators to predict health risk

Gravitational Radiation from Compact Binary Pulsars

An outstanding question in modern Physics is whether general relativity (GR) is a complete description of gravity among bodies at macroscopic scales. Currently, the best experiments supporting this hypothesis are based on high-precision timing of radio pulsars. This chapter reviews recent advances in the field with a focus on compact binary millisecond pulsars with white-dwarf (WD) companions. These systems - if modeled properly - provide an unparalleled test ground for physically motivated alternatives to GR that deviate significantly in the strong-field regime. Recent improvements in observational techniques and advances in our understanding of WD interiors have enabled a series of precise mass measurements in such systems. These masses, combined with high-precision radio timing of the pulsars, result to stringent constraints on the radiative properties of gravity, qualitatively very different from what was available in the past.Comment: Short review chapter to appear in "Gravitational Wave Astrophysics" by Springer-Verlag, edited by Carlos F. Sopuerta; v3: a few major corrections and updated references. Comments are welcome

Multi-laboratory survey of qPCR enterococci analysis method performance in U.S. coastal and inland surface waters

Quantitative polymerase chain reaction (qPCR) has become a frequently used technique for quantifying enterococci in recreational surface waters, but there are several methodological options. Here we evaluated how three method permutations, type of mastermix, sample extract dilution and use of controls in results calculation, affect method reliability among multiple laboratories with respect to sample interference. Multiple samples from each of 22 sites representing an array of habitat types were analyzed using EPA Method 1611 and 1609 reagents with full strength and five-fold diluted extracts. The presence of interference was assessed three ways: using sample processing and PCR amplifications controls; consistency of results across extract dilutions; and relative recovery of target genes from spiked enterococci in water sample compared to control matrices with acceptable recovery defined as 50 to 200%. Method 1609, which is based on an environmental mastermix, was found to be superior to Method 1611, which is based on a universal mastermix. Method 1611 had over a 40% control assay failure rate with undiluted extracts and a 6% failure rate with diluted extracts. Method 1609 failed in only 11% and 3% of undiluted and diluted extracts analyses. Use of sample processing control assay results in the delta-delta Ct method for calculating relative target gene recoveries increased the number of acceptable recovery results. Delta-delta tended to bias recoveries from apparent partially inhibitory samples on the high side which could help in avoiding potential underestimates of enterococci - an important consideration in a public health context. Control assay and delta-delta recovery results were largely consistent across the range of habitats sampled, and among laboratories. The methodological option that best balanced acceptable estimated target gene recoveries with method sensitivity and avoidance of underestimated enterococci densities was Method 1609 without extract dilution and using the delta-delta calculation method. The applicability of this method can be extended by the analysis of diluted extracts to sites where interference is indicated but, particularly in these instances, should be confirmed by augmenting the control assays with analyses for target gene recoveries from spiked target organisms

Risk Characterization of Microplastics in San Francisco Bay, California

Assessing microplastics risk to aquatic ecosystems has been limited by lack of holistic exposure data and poor understanding of biological response thresholds. Here we take advantage of two recent advances, a toxicological meta-analysis that produced biotic response thresholds and a method to quantitatively correct exposure data for sampling methodology biases, to assess microplastic exposure risk in San Francisco Bay, California, USA. Using compartment-specific particle size abundance data, we rescaled empirical surface water monitoring data obtained from manta trawls (> 333 μm) to a broader size (1 to 5000 μm) range, corrected for biases in fiber undercounting and spectroscopic subsampling, and assessed the introduced uncertainty using probabilistic methods. We then compared these rescaled concentrations to four risk thresholds developed to inform risk management for California for each of two effect categories/mechanisms - tissue translocation-mediated effects and food dilution - each aligned to ecologically relevant dose metrics of surface area and volume, respectively. More than three-quarters of samples exceeded the most conservative food dilution threshold, which rose to 85% when considering just the Central Bay. Within the Central Bay, 38% of the samples exceeded a higher threshold associated with management planning, which was statistically significant at the 95% confidence interval. For tissue translocation-mediated effects, no samples exceeded any threshold with statistical significance. The risk associated with food dilution is higher than that found in other systems, which likely reflects this study having been conducted for an enclosed water body. A sensitivity analysis indicated that the largest contributor to assessment variability was associated with estimation of ambient concentration exposure due to correcting for fiber undercounting. Even after compensating for biases associated with fibers and other small particles, concentrations from the trawl samples were still significantly lower than the 1-L grab samples taken at the same time, suggesting our SFB risk estimates are an underestimate. We chose to rely on the trawl data because the 1-L grab sample volume was too small to provide accurate spatial representation, but future risk characterization studies would be improved by using in-line filtration pumps that sample larger volumes while capturing a fuller range of particle size than a towed net

Technical review and evaluation of thermal effect studies and cooling water intake structure demonstration of impact for the Oyster Creek Nuclear Generating Station: Revised final report

The purpose of the final report is to summarize the findings and conclusions of Versar's review and evaluation of the Oyster Creek 316 Demonstration and to make recommendations that assist NJDEP in making a 316 decision for the Oyster Creek NGS, including identifying intake technologies for minimizing environmental harm in preparation for a public hearing. In addition, Versar makes recommendations concerning GPUN's request for special water quality standards for Oyster Creek.May 198