Toxic effect and physiological disruption of sodium phosphate to the quagga mussel (Dreissena bugensis).

Phosphorous is an essential nutrient for all forms of life; however, the question of toxicity to aquatic species remains largely unanswered, despite many systems that exceed natural phosphorus loads. This study determined the ecotoxicological threshold concentration of phosphorus to the freshwater bivalve Dreissena bugensis using a 96-h bioassay. Sublethal, medial lethal, and lethal levels of sodium phosphate to D. bugensis were found to be 125, 260, and 476 ppm. Physiological biomarkers such as the oxygen consumption and filtration rate were estimated by exposing D. bugensis to five different sublethal concentrations (25, 50, 75, 100, and 125 ppm) of sodium phosphate for 96 h. Both oxygen consumption and filtration rate gradually declined with increasing exposure concentrations and durations, which was significant (α < 0.05) for 75, 100, and 125 ppm of sodium phosphate concentrations. Based on the feeding rate and oxygen consumption endpoints, the no-observed effect concentration and the low observed effect concentration were 25 and 75 ppm, respectively. Maximum acceptable toxicant concentration of sodium phosphate was 43.3 ppm. Measured environmental concentration (MEC) of total phosphorus (0.015 ppm; n = 6) was obtained from seasonal field assessments in Saginaw Bay during the years 2008 to 2010. An assessment factor of 1000 was used for calculating the predicted no effect concentration (PNEC) of 0.025 ppm. Risk quotient (RQ) of "0.6" was therefore established using MEC/PNEC (real risk) ratio. Binary ecological classification (RQ < 1) suggested that there is no appreciable risk of phosphorus to D. bungensis in the Saginaw Bay of Lake Huron of Laurentian Great Lakes.

Microbes in Beach Sands: Integrating Environment, Ecology and Public Health.

Beach sand is a habitat that supports many microbes, including viruses, bacteria, fungi and protozoa (micropsammon). The apparently inhospitable conditions of beach sand environments belie the thriving communities found there. Physical factors, such as water availability and protection from insolation; biological factors, such as competition, predation, and biofilm formation; and nutrient availability all contribute to the characteristics of the micropsammon. Sand microbial communities include autochthonous species/phylotypes indigenous to the environment. Allochthonous microbes, including fecal indicator bacteria (FIB) and waterborne pathogens, are deposited via waves, runoff, air, or animals. The fate of these microbes ranges from death, to transient persistence and/or replication, to establishment of thriving populations (naturalization) and integration in the autochthonous community. Transport of the micropsammon within the habitat occurs both horizontally across the beach, and vertically from the sand surface and ground water table, as well as at various scales including interstitial flow within sand pores, sediment transport for particle-associated microbes, and the large-scale processes of wave action and terrestrial runoff. The concept of beach sand as a microbial habitat and reservoir of FIB and pathogens has begun to influence our thinking about human health effects associated with sand exposure and recreational water use. A variety of pathogens have been reported from beach sands, and recent epidemiology studies have found some evidence of health risks associated with sand exposure. Persistent or replicating populations of FIB and enteric pathogens have consequences for watershed/beach management strategies and regulatory standards for safe beaches. This review summarizes our understanding of the community structure, ecology, fate, transport, and public health implications of microbes in beach sand. It concludes with recommendations for future work in this vastly under-studied area.

Performance of viruses and bacteriophages for fecal source determination in a multi-laboratory, comparative study.

An inter-laboratory study of the accuracy of microbial source tracking (MST) methods was conducted using challenge fecal and sewage samples that were spiked into artificial freshwater and provided as unknowns (blind test samples) to the laboratories. The results of the Source Identification Protocol Project (SIPP) are presented in a series of papers that cover 41 MST methods. This contribution details the results of the virus and bacteriophage methods targeting human fecal or sewage contamination. Human viruses used as source identifiers included adenoviruses (HAdV), enteroviruses (EV), norovirus Groups I and II (NoVI and NoVII), and polyomaviruses (HPyVs). Bacteriophages were also employed, including somatic coliphages and F-specific RNA bacteriophages (FRNAPH) as general indicators of fecal contamination. Bacteriophage methods targeting human fecal sources included genotyping of FRNAPH isolates and plaque formation on bacterial hosts Enterococcus faecium MB-55, Bacteroides HB-73 and Bacteroides GB-124. The use of small sample volumes (≤50 ml) resulted in relatively insensitive theoretical limits of detection (10-50 gene copies or plaques × 50 ml(-1)) which, coupled with low virus concentrations in samples, resulted in high false-negative rates, low sensitivity, and low negative predictive values. On the other hand, the specificity of the human virus methods was generally close to 100% and positive predictive values were ∼40-70% with the exception of NoVs, which were not detected. The bacteriophage methods were generally much less specific toward human sewage than virus methods, although FRNAPH II genotyping was relatively successful, with 18% sensitivity and 85% specificity. While the specificity of the human virus methods engenders great confidence in a positive result, better concentration methods and larger sample volumes must be utilized for greater accuracy of negative results, i.e. the prediction that a human contamination source is absent.

Recommendations following a multi-laboratory comparison of microbial source tracking methods.

Microbial source tracking (MST) methods were evaluated in the Source Identification Protocol Project (SIPP), in which 27 laboratories compared methods to identify host sources of fecal pollution from blinded water samples containing either one or two different fecal types collected from California. This paper details lessons learned from the SIPP study and makes recommendations to further advance the field of MST. Overall, results from the SIPP study demonstrated that methods are available that can correctly identify whether particular host sources including humans, cows and birds have contributed to contamination in a body of water. However, differences between laboratory protocols and data processing affected results and complicated interpretation of MST method performance in some cases. This was an issue particularly for samples that tested positive (non-zero Ct values) but below the limits of quantification or detection of a PCR assay. Although false positives were observed, such samples in the SIPP study often contained the fecal pollution source that was being targeted, i.e., the samples were true positives. Given these results, and the fact that MST often requires detection of targets present in low concentrations, we propose that such samples be reported and identified in a unique category to facilitate data analysis and method comparisons. Important data can be lost when such samples are simply reported as positive or negative. Actionable thresholds were not derived in the SIPP study due to limitations that included geographic scope, age of samples, and difficulties interpreting low concentrations of target in environmental samples. Nevertheless, the results of the study support the use of MST for water management, especially to prioritize impaired waters in need of remediation. Future integration of MST data into quantitative microbial risk assessments and other models could allow managers to more efficiently protect public health based on site conditions.

Differentiating Enterococcus concentration spatial, temporal, and analytical variability in recreational waters.

Monitoring recreational waters for fecal contamination is an important responsibility of water resource management agencies throughout the world, yet fecal indicator bacteria (FIB)-based recreational water quality assessments rarely distinguish between analytical, spatial, and temporal variability. To address this gap in water resources research and management protocol, we compare two methods for quantifying FIB concentration variability at a frequently-used beach on Lake Huron (Michigan, USA). The first method calculates differences between most probable number (MPN) and colony-forming unit (CFU) values derived from conventional analysis procedures. The second method uses the "raw data" from these analysis procedures in a Bayesian hierarchical model to explicitly acknowledge analytical variability and subsequently infer the relative significance of the effect of sampling location and time on in situ FIB concentrations. Results of the Bayesian analysis indicate that in situ FIB concentrations do not vary significantly over small spatial and temporal scales, and that observed differences in MPN and CFU values over these same spatial and temporal scales are due almost entirely to intrinsic variability introduced by laboratory analysis procedures. Our findings underscore potential opportunities for incorporating Bayesian statistical models directly into routine recreational water quality assessments and for advancing the state of the art in methods for protecting humans from waterborne disease.

In vitro antioxidant and antimicrobial activities of two Hawaiian marine Limu: Ulva fasciata (Chlorophyta) and Gracilaria salicornia (Rhodophyta).

The antioxidant and antimicrobial properties of two Hawaiian marine algae (Ulva fasciata and Gracilaria salicornia) were evaluated. Ethanolic extracts of these two algae exhibited intracellular reactive 1,1-diphenyl-2-picrylhydrazyl, 2,2'-azino-bis(3-ethylbenzthiazoline)-6-sulfonic radical, nitric oxide, superoxide radical, and hydroxyl radical scavenging activities. In addition to the antioxidant activity these algae possessed appreciable antimicrobial activity and total phenolic contents. The overall results have established that these two marine algae could be used to derive bioactive compounds for a possible role as nutraceutical agents.

Isolation and characterization of Bacteroides host strain HB-73 used to detect sewage specific phages in Hawaii.

Previous studies have shown that Escherichia coli and enterococci are unreliable indicators of fecal contamination in Hawaii because of their ability to multiply in environmental soils. In this study, the method of detecting Bacteroides phages as specific markers of sewage contamination in Hawaii's recreational waters was evaluated because these sewage specific phages cannot multiply under environmental conditions. Bacteroides hosts (GB-124, GA-17), were recovered from sewage samples in Europe and were reported to be effective in detecting phages from sewage samples obtained in certain geographical areas. However, GB-124 and GA-17 hosts were ineffective in detecting phages from sewage samples obtained in Hawaii. Bacteroides host HB-73 was isolated from a sewage sample in Hawaii, confirmed as a Bacteroides sp. and shown to recover phages from multiple sources of sewage produced in Hawaii at high concentrations (5.2-7.3 x 10(5) PFU/100 mL). These Bacteroides phages were considered as potential markers of sewage because they also survived for three days in fresh stream water and two days in marine water. Water samples from Hawaii's coastal swimming beaches and harbors, which were known to be contaminated with discharges from streams, were shown to contain moderate (20-187 CFU/100 mL) to elevated (173-816 CFU/100 mL) concentrations of enterococci. These same samples contained undetectable levels (<10 PFU/100 mL) of F+ coliphage and Bacteroides phages and provided evidence to suggest that these enterococci may not necessarily be associated with the presence of raw sewage. These results support previous conclusions that discharges from streams are the major sources of enterococci in coastal waters of Hawaii and the most likely source of these enterococci is from environmental soil rather than from sewage.

Water chemistry influences the toxicity of silver to the green-lipped mussel Perna viridis.

The study determined the influence and relative importance of water chemistry parameters (pH, alkalinity, hardness) on the acute toxicity of silver to the green mussel Perna viridis. A preliminary bioassay revealed that 4 mg L(-1) of silver caused 50% mortality (LC50) in 96 h for mussels placed in seawater with pH 8.5, hardness 1,872 mg L(-1), and alkalinity 172 mg L(-1). Mortality of mussels increased with decreasing pH and increasing hardness and alkalinity variables. In contrast the mortality decreased with increasing pH and decreasing hardness and alkalinity values. The water chemistry also affected the concentration of silver in experimental seawater and bioaccumulation of silver in mussels. The results revealed that the chemical properties of seawater must be considered while conducting toxicity tests with metals like silver. The possible explanations for the influence of water chemistry on silver toxicity to P. viridis are discussed.