Persistence of extracellular DNA in river sediment facilitates antibiotic resistance gene propagation.

The propagation of antibiotic resistance genes (ARGs) represents a global threat to both human health and food security. Assessment of ARG reservoirs and persistence is therefore critical for devising and evaluating strategies to mitigate ARG propagation. This study developed a novel, internal standard method to extract extracellular DNA (eDNA) and intracellular DNA (iDNA) from water and sediments, and applied it to determine the partitioning of ARGs in the Haihe River basin in China, which drains an area of intensive antibiotic use. The concentration of eDNA was higher than iDNA in sediment samples, likely due to the enhanced persistence of eDNA when associated with clay particles and organic matter. Concentrations of sul1, sul2, tetW, and tetT antibiotic resistance genes were significantly higher in sediment than in water, and were present at higher concentrations as eDNA than as iDNA in sediment. Whereas ARGs (frequently located on plasmid DNA) were detected for over 20 weeks, chromosomally encoded 16S rRNA genes were undetectable after 8 weeks, suggesting higher persistence of plasmid-borne ARGs in river sediment. Transformation of indigenous bacteria with added extracellular ARG (i.e., kanamycin resistance genes) was also observed. Therefore, this study shows that extracellular DNA in sediment is a major ARG reservoir that could facilitate antibiotic resistance propagation.

Addition of a magnetite layer onto a polysulfone water treatment membrane to enhance virus removal.

The applicability of low-pressure membranes systems in distributed (point of use) water treatment is hindered by, among other things, their inability to remove potentially harmful viruses and ions via size exclusion. According to the USEPA and the Safe Drinking Water Act, drinking water treatment processes must be designed for 4-log virus removal. Batch experiments using magnetite nanoparticle (nano-Fe3O4) suspensions and water filtration experiments with polysulfone membranes coated with nano-Fe3O4 were conducted to assess the removal of a model virus (bacteriophage MS2). The membranes were coated via a simple filtration protocol. Unmodified membranes were a poor adsorbent for MS2 bacteriophage with less than 0.5-log removal, whereas membranes coated with magnetite nanoparticles exhibited a removal efficiency exceeding 99.99% (4-log). Thus, a cartridge of PSf membranes coated with nano-Fe3O4 particles could be used to remove viruses from water. Such membranes showed negligible iron leaching into the filtrate, thus obviating concern about coloured water. Further research is needed to reduce the loss of water flux caused by coating.

Implications and potential applications of bactericidal fullerene water suspensions: effect of nC(60) concentration, exposure conditions and shelf life.

Stable fullerene water suspensions (nC(60)) exhibited potent antibacterial activity to physiologically different bacteria in low-salts media over a wide range of exposure conditions. Antibacterial activity was observed in the presence or absence of light or oxygen, and increased with both exposure time and dose. The activity was also influenced by the nC(60) storage conditions and by the age of the buckminsterfullerene (C(60)) used to make nC(60). These results reflect the potential impact of nC(60) on the health of aquatic ecosystems and suggest novel alternatives for disinfection and microbial control.

Comparison of bioaugmentation and biostimulation for the enhancement of dense nonaqueous phase liquid source zone bioremediation.

Two 11.7-m(3) experimental controlled release systems (ECRS), packed with sandy model aquifer material and amended with tetrachloroethene (PCE) dense nonaqueous phase liquid (DNAPL) source zone, were operated in parallel with identical flow regimes and electron donor amendments. Hydrogen Releasing Compound (Regenesis Bioremediation Products, Inc., San Clemente, California), and later dissolved lactate, served as electron donors to promote dechlorination. One ECRS was bioaugmented with an anaerobic dechlorinating consortium directly into the source zone, and the other served as a control (biostimulated only) to determine the benefits of bioaugmentation. The presence of halorespiring bacteria in the aquifer matrix before bioaugmentation, shown by nested polymerase chain reaction with phylogenetic primers, suggests that dechlorinating catabolic potential may be somewhat widespread. Results obtained corroborate that source zone reductive dechlorination of PCE is possible at near field scale and that a system bioaugmented with a competent halorespiring consortium can enhance DNAPL dissolution and dechlorination processes at significantly greater rates than in a system that is biostimulated only.

Transport of antibiotic-resistant bacteria and resistance-carrying plasmids through porous media.

Microbial antibiotic resistance has emerged as not only a major medical problem but also as an environmental engineering challenge, with antibiotic resistance genetic elements as environmental pollutants. The transport characteristics of a tetracycline-resistant bacterium (B. cepacia) and plasmids carrying tetracycline-resistance genes were investigated using flow-through columns packed with porous media. Higher influent cell concentrations (1.1 x 10(8) CFU mL(-1)) resulted in higher breakthrough (C/Co = 0.596+/-0.055) than a solution with lower cell concentration (2.0 x 10(6) CFU mL(- 1), C/Co = 0.461+/-0.037). This decreased extent of filtration suggests fast initial cell deposition and strong subsequent blocking of binding sites, resulting in less-hindered microbial transport through the sandy medium. The addition of a bromide tracer (NaBr) prior to the plasmid solution resulted in DNA retardation and increased filtration in a zirconia-silica bead matrix. Apparently, Na+ binding to the beads decreased electrostatic repulsion between the negatively charged DNA and zirconia-silica surface. In contrast, plasmid breakthrough preceded that of the tracer when the plasmids were added first, possibly due to size exclusion chromatography coupled with stronger electrostatic repulsion. This implies that efforts to characterize the dynamics of resistance vector propagation in aquifers should consider the effect of groundwater chemistry and the surface characteristics of the porous media on vector transport.

Comparative toxicity of nano-scale TiO2, SiO2 and ZnO water suspensions.

TiO2, SiO2 and ZnO are common additives with improved applications at the nanoscale. The antibacterial activity of TiO2, which has important ecosystem health implications, is well understood. However, less attention has been paid to the antibacterial activity of SiO2 and ZnO despite them also producing reactive oxygen species. This paper explores the relative toxicity of TiO2, SiO2 and ZnO water suspensions towards bacteria (B. subtilis, E. coli) and the eukaryotic Daphnia magna. These three photosensitive nanomaterials were hazardous to all test organisms, with toxicity increasing with particle concentration. Toxicity of the three compounds decreased from ZnO to TiO2 to SiO2 and Daphnia were most susceptible to their effects. Nominal particle size did not affect the toxicity of these compounds. Antibacterial activity was noted under both dark and light conditions indicating that mechanisms additional to ROS production were responsible for growth inhibition. These results highlight the need for caution during the use and disposal of such manufactured nanomaterials to prevent unintended environmental impacts, as well as the importance of further research on the mechanisms and factors that increase toxicity to enhance risk management.

Phenanthrene removal by Penicillium frequentans grown on a solid-state culture: effect of oxygen concentration.

Phenanthrene removal by Penicillium frequentans was compared under aerobic and microaerophilic conditions in a solid culture amended with low quantities of an agricultural residue. An inoculum of P. frequentans grown on sugarcane bagasse pith was mixed with soil spiked with 200 mg l(-1) of phenanthrene, to obtain a final bagasse/soil ratio of 1:16. The C/N ratio was adjusted to 60 and the moisture content to 40%. The oxygen concentrations were adjusted to 20%, 10%, 5%, 2% and close to 0%, in the soil-gas phase for each treatment. There were statistically significant (p<0.05) differences in the metabolic activity at different oxygen concentrations, measured as CO2 production. Phenanthrene removal rates increased with oxygen concentration, reaching 52% removal after 17 days of incubation for the treatment with 20% O2. Nevertheless, oxygen-limited (microaerophilic) conditions did not preclude phenanthrene degradation.

Prevalence and proliferation of antibiotic resistance genes in two municipal wastewater treatment plants.

The propagation of antibiotic resistance genes (ARGs) is an emerging health concern worldwide. Thus, it is important to understand and mitigate their occurrence in different systems. In this study, 30 ARGs that confer resistance to tetracyclines, sulfonamides, quinolones or macrolides were detected in two activated sludge wastewater treatment plants (WWTPs) in northern China. Bacteria harboring ARGs persisted through all treatment units, and survived disinfection by chlorination in greater percentages than total Bacteria (assessed by 16S rRNA genes). Although the absolute abundances of ARGs were reduced from the raw influent to the effluent by 89.0%-99.8%, considerable ARG levels [(1.0 ± 0.2) × 10(3) to (9.5 ± 1.8) × 10(5) copies/mL)] were found in WWTP effluent samples. ARGs were concentrated in the waste sludge (through settling of bacteria and sludge dewatering) at (1.5 ± 2.3) × 10(9) to (2.2 ± 2.8) × 10(11) copies/g dry weight. Twelve ARGs (tetA, tetB, tetE, tetG, tetH, tetS, tetT, tetX, sul1, sul2, qnrB, ermC) were discharged through the dewatered sludge and plant effluent at higher rates than influent values, indicating overall proliferation of resistant bacteria. Significant antibiotic concentrations (2%-50% of raw influent concentrations) remained throughout all treatment units. This apparently contributed selective pressure for ARG replication since the relative abundance of resistant bacteria (assessed by ARG/16S rRNA gene ratios) was significantly correlated to the corresponding effluent antibiotic concentrations. Similarly, the concentrations of various heavy metals (which induce a similar bacterial resistance mechanism as antibiotics - efflux pumps) were also correlated to the enrichment of some ARGs. Thus, curtailing the release of antibiotics and heavy metals to sewage systems (or enhancing their removal in pre-treatment units) may alleviate their selective pressure and mitigate ARG proliferation in WWTPs.

A model for the effect of rhizodeposition on the fate of phenanthrene in aged contaminated soil.

Microcosm data were used to develop a deterministic model to describe how rhizodeposition affects the fate of phenanthrene in aged contaminated soil. Microbial mineralization and soil sequestration of 14C-phenanthrene were compared in microcosms amended weekly with phenolic-rich mulberry root extracts versus unamended controls. Mineralization was higher in the amended soils simulating the rhizosphere (57.7 +/- 0.9%) than in controls simulating bulk (unplanted) soils (53.2 +/- 0.7%) after 201 days (p < 0.05). Humin was the main soil sink for the residual 14C-label. Whereas the total 14C-label associated with humin remained constant in biologically active soils (at about 30%), it increased up to 80% after 201 days in sterile controls. The initial phenanthrene extraction with n-butanol (commonly used to assess bioavailability) slightly underestimated the fraction thatwas mineralized (assessed by 14CO2 recovery). Changes in the unextractable fraction (determined by combustion in a biological oxidizer) suggested the presence of two soil sequestration domains: (1) irreversibly bound residue, and (2) an intermediate transition phase that is unextractable by solvents at a given point in time but could become bioavailable due to physicochemical or biological transformations of the binding matrix. The fate of phenanthrene was accurately modeled by considering the transfer of the 14C label between different soil compartments as first-order kinetic processes. Model simulations suggested that the system was approaching a stable end-point after 201 days of simulated rhizoremediation, and corroborated that microorganisms have a significant impact on the fate of phenanthrene in soil.