Improved traceability of Shiga-toxin-producing <Emphasis Type="Italic">Escherichia coli</Emphasis> using CRISPRs for detection and typing

Among strains of Shiga-toxin-producing Escherichia coli (STEC), seven serogroups (O26, O45, O103, O111, O121, O145, and O157) are frequently associated with severe clinical illness in humans. The development of methods for their reliable detection from complex samples such as food has been challenging thus far, and is currently based on the PCR detection of the major virulence genes stx1, stx2, and eae, and O-serogroup-specific genes. However, this approach lacks resolution. Moreover, new STEC serotypes are continuously emerging worldwide. For example, in May 2011, strains belonging to the hitherto rarely detected STEC serotype O104:H4 were identified as causative agents of one of the world’s largest outbreak of disease with a high incidence of hemorrhagic colitis and hemolytic uremic syndrome in the infected patients. Discriminant typing of pathogens is crucial for epidemiological surveillance and investigations of outbreaks, and especially for tracking and tracing in case of accidental and deliberate contamination of food and water samples. Clustered regularly interspaced short palindromic repeats (CRISPRs) are composed of short, highly conserved DNA repeats separated by unique sequences of similar length. This distinctive sequence signature of CRISPRs can be used for strain typing in several bacterial species including STEC. This review discusses how CRISPRs have recently been used for STEC identification and typing.     

Distribution of antidepressants and their metabolites in brook trout exposed to municipal wastewaters before and after ozone treatment--evidence of biological effects

This study examined the tissues distribution of selected serotonin reuptake inhibitors (SSRIs) in brook trout exposed for 3 months to continuous flow-through primary-treated effluent before and after ozone treatment. A reliable analytical method was developed for the quantification of trace amounts of antidepressants in small tissue homogenate extracts. Levels of six antidepressants and four of their N-desmethyl metabolites were determined using liquid chromatography-tandem mass spectrometry. Significant amounts of the SSRIs were found in fish tissue-in decreasing order: liver>brain>muscle. Sertraline and its metabolite desmethylsertraline were the predominant substances observed in most tissues (0.04-10.3 ng g(-1)). However, less SSRIs (0.08-1.17 ng g(-1)) were bioaccumulated in the ozonated effluent. The early molecular effects of these SSRIs on the Na/K-dependent ATPase pump activity in brain synaptosomes where also investigated in vitro and in fish exposed to the municipal effluents. With respect to their potential biological effects, in vitro exposure to selected SSRIs induced a reduction of the brain Na/K-ATPase activity in synaptosomes in a dose-dependent manner. Results showed that Na/K-ATPase activity was readily inhibited by exposure to municipal effluent before and, to a lesser extent, after ozone treatment. Moreover, the Na/K-ATPase activity was significantly and negatively correlated with brain tissue concentrations of fluoxetine (r=-0.57; p<0.03), desmethylsertraline (r=-0.84; p<0.001), and sertraline (r=-0.82; p<0.001). The present study reveals that SSRIs are readily available in fish, biologically active and corroborates previous findings on the serotonergic properties of municipal effluents to aquatic organisms.     

Distinct diversity of the czcA gene in two sedimentary horizons from a contaminated estuarine core

In estuarine ecosystems, trace metals are mainly associated with fine grain sediments which settle on mudflats. Over time, the layers of sediments accumulate and are then transformed by diagenetic processes, recording the history of the estuary’s chemical contamination. In such a specific environment, we investigated to what extent a chronic exposure to contaminants could affect metal-resistant sedimentary bacteria in subsurface sediments. The occurrence and diversity of cadmium resistance genes (cadA, czcA) was investigated in 5- and 33-year-old sediments from a highly contaminated estuary (Seine France). Primers were designed to detect a 252-bp fragment of the czcA gene, specifically targeting a transmembrane helice domain (TMH IV) involved in the proton substrate antiport of this efflux pump. Although the cadA gene was not detected, the highest diversity of the sequence of the czcA gene was observed in the 5-year-old sediment. According to the percentage of identity at the amino acid level, the closest CzcA relatives were identified among Proteobacteria (α, β, γ, and δ), Verrucomicrobia, Nitrospirae, and Bacteroidetes. The most abundant sequences were affiliated with Stenotrophomonas. In contrast, in the 33-year-old sediment, CzcA sequences were mainly related to Rhodanobacter thiooxydans and Stenotrophomonas, suggesting a shaping of the metal-resistant microbial communities over time by both diagenetic processes and trace metal contamination.     

Use of AERMOD to Determine a Hydrogen Sulfide Emission Factor for Swine Operations by Inverse Modeling

This study was conducted to determine both optimal settings applied to the plume dispersion model, AERMOD, and a scalable emission factor for accurately determining the spatial distribution of hydrogen sulfide concentrations in the vicinity of swine concentrated animal feeding operations (CAFOs). These operations emit hydrogen sulfide from both housing structures and waste lagoons. With ambient measurements made at 4 stations within 1 km of large swine CAFOs in Iowa, an inverse-modeling approach applied to AERMOD was used to determine hydrogen sulfide emission rates. CAFO buildings were treated as volume sources whereas nearby lagoons were modeled as area sources. The robust highest concentration (RHC), calculated for both measured and modeled concentrations, was used as the metric for adjusting the emission rate until the ratio of the two RHC levels was unity. Utilizing this approach, an average emission flux rate of 0.57 μg/m(2)-s was determined for swine CAFO lagoons. Using the average total animal weight (kg) of each CAFO, an average emission factor of 6.06 × 10(-7) μg/yr-m(2)-kg was calculated. From studies that measured either building or lagoon emission flux rates, building fluxes, on a floor area basis, were considered equal to lagoon flux rates. The emission factor was applied to all CAFOs surrounding the original 4 sites and surrounding an additional 6 sites in Iowa, producing an average modeled-to-measured RHC ratio of 1.24. When the emission factor was applied to AERMOD to simulate the spatial distribution of hydrogen sulfide around a hypothetical large swine CAFO (1M kg), concentrations 0.5 km from the CAFO were 35 ppb and dropped to 2 ppb within 6 km of the CAFO. These values compare to a level of 30 ppb that has been determined by the State of Iowa as a threshold level for ambient hydrogen sulfide levels.     

Human visitation rates to the Apostle Islands National Lakeshore and the introduction of the non-native species Lymantria dispar (L.)

The introduction of non-native species has accelerated due to increasing levels of global trade and travel, threatening the composition and function of ecosystems. Upon arrival and successful establishment, biological invaders begin to spread and often do so with considerable assistance from humans. Recreational areas can be especially prone to the problem of accidental non-native species transport given the number of visitors that arrive from geographically diverse areas. In this paper, we examine camping permit data to the Apostle Islands National Lakeshore in northwestern Wisconsin, USA, from 1999 to 2007 relative to gypsy moth distribution, phenology and outbreak data. During this time, gypsy moth populations became established in this area ahead of the moving population front of the gypsy moth, suggesting anthropogenic introduction. The permit data revealed that the majority of visitors arrived from outside of the gypsy moth established area. However, there was a consistent yearly trend of visitors that arrived from areas of high gypsy moth populations and who arrived during the gypsy moth life stage (egg masses) most likely to be successfully introduced. Using available data on the gypsy moth and its relationship to camping permit data, we describe how recreational managers could optimize park strategies to mitigate unwanted introductions of the gypsy moth as well as develop analogous strategies for managing other biological invaders in recreational areas.     

ANALYSIS OF WIND- AND SHIP-INDUCED SEDIMENT RESUSPENSION IN DULUTH-SUPERIOR HARBOR1

ABSTRACT: Suspended solids and ammonium concentration profiles measured at five locations in Duluth-Superior Harbor during July-October 1985 were analyzed to quantify wind and ship effects on sediment resuspension and resulting harbor water quality. Wind components from the SE quadrant correlated strongly with depth-averaged suspended solids concentrations that were unaffected by ship passage or thermal stratification. Winds from that quadrant have the largest fetch in the harbor. The highest correlation (r²= 0.93) was with the 6-hour average of the ESE wind velocity component. Multiple linear regression analysis of data from post-ship passage concentration profiles yielded numerical estimates of settling velocities of 0.08 to 0.25 cm s^?1, typical of ship-resuspended sediments, and vertical eddy diffusivities of 4 to 13 cm² s¹. The results suggest that ambient vertical eddy diffusivities in the harbor are less than 4 cm² s^?1 in the absence of ship passages and with winds less than 5 m s^?1 (10 knots).     

Hypsographs: Basic tools for lake management and monitoring

Hypsographs are graphic depictions of the vertical distributions of volumes or masses of water in lakes, ponds, or reservoirs. Quantitative limnological science or engineering must use a hypsograph, or a more complex equivalent system, to undertake calculations of water constituent concentration data by mathematical weighting. Construction of the graph is simple and described. Exemplary uses of the hypsograph are presented.     

SOME CONJUNCTIVE DESALTING STUDIES EN THE UNITED KINGDOM1

ABSTRACT .Operating rule procedures are developed for the optimal use of desalination in conjunction with surface water impoundments. Results obtained from the use of hill-climbing and dynamic programming methods are compared. Benefits to be obtained from the integrated use of desalination are demonstrated through a case study for the Barcelona area of Spain.     

THE POLITICAL INFLUENCE OF RESIDENTIAL CONSUMERS ON WATER RATES1

ABSTRACT: The regulatory process such as that for water rates can be viewed as a bargaining process between regulators, utility management, and consumers. This bargaining process involves both political and economic influences. In an attempt to empirically verify that utility rates are a product of both economic and political factors, a sample of publicly-owned water utilities was subjected to regression analysis. As expected, cost and cost-related factors were positive and significant determinants of residential water rates. Using an admittedly imperfect measure of residential user political influence, there was evidence that increased political activity results in lower water rates. Furthermore, the political influence and water rate linkage is more evident in small water utilities than in larger suppliers.