Characterization of the cell surface properties of drinking water pathogens by microbial adhesion to hydrocarbon and electrophoretic mobility measurements.

The surface characteristics of microbial cells directly influence their mobility and behavior within aqueous environments. The cell surface hydrophobicity (CSH) and electrophoretic mobility (EPM) of microbial cells impact a number of interactions and processes including aggregation, adhesion to surfaces, and stability of the cells within the aqueous environments. These cell characteristics are unique to the bacterial species and are a reflection of the large diversity of surface structures, proteins, and appendages of microorganisms. CSH and EPM of bacterial cells contribute substantially to the effectiveness of drinking water treatment to remove them, and therefore an investigation of these properties will be useful in predicting their removal through drinking water treatment processes and transport through drinking water distribution systems. EPM and CSH measurements of six microbiological pathogen or surrogate species suspended in phosphate-buffered water are reported in this work. Two strains of Vibrio cholerae were hydrophobic, while three strains of Escherichia coli were hydrophilic. Bacillus cereus was categorized as moderately hydrophobic. The strains of E. coli had the highest (most negative) EPM. Based on the measurements, E. coli species is predicted to be most difficult to remove from water while V. cholerae will be the easiest to remove.

Fate of pentabrominated diphenyl ethers in soil: abiotic sorption, plant uptake, and the impact of interspecific plant interactions.

Polybrominated diphenyl ethers (PBDEs) are potentially harmful and persistent environmental pollutants. Despite evidence that soils are a major sink for PBDEs, little is known regarding their behavior in this medium. An environmentally relevant level of a commercial penta-BDE mixture (75 microg kg(-1)) was added to topsoil, and the extractability of three congeners (BDE-47, -99, and -100) was monitored over 10 weeks in planted and unplanted treatments. The extractability of each congener decreased rapidly in the experimental soil due largely to abiotic sorption to soil particles, which was demonstrated by low PBDE recovery from sterilized and dry soils. Monoculture plantings of zucchini and radish did not affect the recovery of PBDEs from soil. However, PBDE recovery from mixed species plantings was nearly 8 times higher than that of unplanted and monoculture treatments, indicating that interspecific plant interactions may enhance PBDE bioavailablity in soil. Evidence for competitive interactions between the two species was revealed by reduced shoot biomass of zucchini plants in mixed treatments relative to pots containing only zucchini. Both plant species accumulated PBDEs in root and shoot tissue (<5 microg kg(-1) plant tissue). PBDE uptake was higher in zucchini, and translocation of PBDEs to zucchini shoots was congener-specific. Our results suggest that although abiotic sorption may limit the potential for human exposure to PBDEs in soil, plants may increase the exposure risk by taking up and translocating PBDEs into aboveground tissues and by enhancing bioavailability in soil.

Use of SEC-ICP-MS with a collision cell for determining the interaction of chromium with DNA extracted from metal-contaminated soils.

The potential of chromium to bind to DNA isolated directly from soil microbial communities was investigated in this study. An analytical scheme was developed to distinguish between chromium bound to DNA and its fragments or chromium contained elsewhere in an environmental DNA extract. DNA was extracted from chromium-contaminated soils and purified using DNA clean-up resins. Size-exclusion chromatography was employed due to its advantages in the separation and molecular weight approximation of large biomolecules. It was coupled with two on-line detection systems (spectrophotometric and inductively coupled plasma mass spectrometric) to study the binding of chromium to DNA or other components in a DNA extract. A collision cell was pressurized with helium to remove diatomic and polyatomic interferents resulting from the chosen mobile phase. Chromium peaks were observed in both the large and small molecular weight regions of the chromatogram; to further confirm that the environmentally extracted DNA contained Cr, the subsequently purified DNA was examined for total Cr using flow injection ICP-MS to accommodate small sample volumes. DNA samples isolated from the two soils examined contained 0.5-0.7 ppb Cr, indicating that DNA isolated directly from a chromium-contaminated soil has chromium bound to the nucleic acids.

The widespread occurrence of the enterohemolysin gene ehlyA among environmental strains of Escherichia coli.

The putative virulence factor enterohemolysin, encoded by the ehlyA gene, has been closely associated with the pathogenic enterohemorrhagic Escherichia coli (EHEC) group. Escherichia coli isolates from effluents from seven geographically dispersed municipal wastewater treatment plants were screened for the presence of enterohemolysin. A total of 338 E. coli isolates were found to express the ehlyA gene. However, none of the isolates contained the toxin-encoding genes (stxA or stxB) associated with EHEC. Two of the 338 isolates possessed the virulence factor intimin, encoded by the eae gene. These findings suggest that the ehlyA gene may be widely distributed among non-EHEC isolates in the environment.

In-situ enumeration and probing of pyrene-degrading soil bacteria.

Inferences about which microorganisms degrade polycyclic aromatic hydrocarbons in contaminated soils have largely been obtained using culture-based techniques, despite the low percentage of microorganisms in soil that are believed to be culturable. We used a substrate-responsive direct viable count method to identify and quantify potential polycyclic aromatic hydrocarbon-degrading bacteria in a soil containing petroleum wastes. Bacteria were extracted and their response to substrates determined in the presence of DNA gyrase inhibitors, which cause viable and active cells to elongate. When yeast extract, a widely used carbon source, was added as a growth substrate, together with nalidixic acid, piromidic acid and ciprofloxacin, a significant increase in elongated cells to 47%, 37% and 22%, respectively, was observed within 24 h. With pyrene as the main substrate, 10 mg L(-1) of nalidixic acid or piromidic acid caused 18-22% and 8-12%, respectively, of the cells to elongate within 24 h; whereas the effect of 0.5 mg L(-1) ciprofloxacin was not significant until 53 h later. Enlarged cells were identified and enumerated by fluorescent in situ hybridization, using Alpha-, Beta- and Gammaproteobacteria, and domain Bacteria-specific probes. The Bacteria-specific probe detected 35-71% of the total microorganisms detected by the DNA-binding dye 4,6-diamidino-2-phenylindole. Initially, 44%, 13% and 5% of the total bacteria in the soil extract were Alpha-, Beta- and Gammaproteobacteria, respectively. Without pyrene or a gyrase inhibitor, these subgroups decreased to 30% of the total population but were predominant with piromidic acid or unchanged with ciprofloxacin when pyrene was the main substrate. The proportion of elongated Alpha- and Betaproteobacteria (potential pyrene degraders) increased significantly (P<0.05). This approach links phylogenetic information with physiological function in situ without the conventional cultivation of bacteria and can be used to probe and enumerate degradative groups at even a finer level of discrimination.

High performance degradation of azo dye Acid Orange 7 and sulfanilic acid in a laboratory scale reactor after seeding with cultured bacterial strains.

Bacterial strains 1CX and SAD4i--previously isolated from the mixed liquor of a municipal sewage treatment plant--are capable of degrading the azo dye Acid Orange 7 (AO7) and sulfanilic acid, respectively. A rotating drum bioreactor (RDBR), operating under continuous flow and nutrient conditions designed to simulate the effluent from a dye manufacturing plant, was seeded with strains 1CX and SAD4i, forming a biofilm capable of degrading AO7 and sulfanilic acid. In addition, an RDBR containing a pre-existing biofilm capable of degrading AO7, but not sulfanilic acid, was seeded with strain SAD4i alone. Strain SAD4i was incorporated into the existing biofilm and degraded the sulfanilic acid resulting from the degradation of AO7 by indigenous members of the biofilm. The ability to seed a bioreactor with bacterial strains capable of degrading azo dyes, and resulting by-products, in a mixed microbial community suggests that this process could have commercial applications.

Degradation of acid orange 7 in an aerobic biofilm.

A stable microbial biofilm community capable of completely mineralizing the azo dye acid orange 7 (AO7) was established in a laboratory scale rotating drum bioreactor (RDBR) using waste liquor from a sewage treatment plant. A broad range of environmental conditions including pH (5.8-8.2), nitrification (0.0-4.0 mM nitrite), and aeration (0.2-6.2 mg O2 l(-1)) were evaluated for their effects on the biodegradation of AO7. Furthermore the biofilm maintained its biodegradative ability for over a year while the effects of these environmental conditions were evaluated. Reduction of the azo bond followed by degradation of the resulting aromatic amine appears to be the mechanism by which this dye is biodegraded. Complete loss of color, sulfanilic acid, and chemical oxygen demand (COD) indicate that AO7 is mineralized. To our knowledge this is the first reported occurrence of a sulfonated phenylazonaphthol dye being completely mineralized under aerobic conditions. Two bacterial strains (ICX and SAD4i) originally isolated from the RDBR were able to mineralize, in co-culture, up to 90% of added AO7. During mineralization of AO7, strain ICX reduces the azo bond under aerobic conditions and consumes the resulting cleavage product 1-amino-2-naphthol. Strain SAD4i consumes the other cleavage product, sulfanilic acid. The ability of the RDBR biofilm to aerobically mineralize an azo dye without exogenous carbon and nitrogen sources suggests that this approach could be used to remediate industrial wastewater contaminated with spent dye.