Ultra-fine-ZnO showed low toxicity in complex water matrix containing multiple components such as PBS buffer and the toxic mechanism of ultra-fine-ZnO has not been clearly elucidated. In present study, enhanced antibacterial activity of 200 nm diameter ultra-fine-ZnO in PBS buffer against Bacillus cereus and Escherichia coli were observed in the presence of several organic acids in comparison with ultra-fine-ZnO in PBS buffer alone. These findings indicated that the toxic effects of the ultra-fine-ZnO was dependent on the concentration of released Zn2+ which was affected by organic acids. The production of reactive oxygen species (ROS) did not responsible to the toxic mechanism of ultra-fine-ZnO which was tested using the antioxidant N-Acetylcysteine (NAC). Indeed, ultra-fine-ZnO induced bacteria cell membrane leakages and cell morphology damages that eventually led to cell death, which were confirmed using propidium monoazide (PMA) in combination with PCR and scanning electron microscopy (SEM). All data gathered herein suggested that released Zn2+ played a major role in the microbial toxicity of ultra-fine-ZnO.

The objective of this study was to determine the prevalence of Staphylococcus in urban wastewater treatment plants (UWTP) of La Rioja (Spain), and to characterize de obtained isolates. 16 wastewater samples (8 influent, 8 effluent) of six UWTPs were seeded on mannitol-salt-agar and oxacillin-resistance-screening-agar-base for staphylococci and methicillin-resistant Staphylococcus aureus recovery. Antimicrobial susceptibility profile was determined for 16 antibiotics and the presence of 35 antimicrobial resistance genes and 14 virulence genes by PCR. S. aureus was typed by spa, agr, and multilocus-sequence-typing, and the presence of immune-evasion-genes cluster was analyzed. Staphylococcus spp. were detected in 13 of 16 tested wastewater samples (81%), although the number of CFU/mL decreased after treatment. 40 staphylococci were recovered (1–5/sample), and 8 of them were identified as S. aureus being typed as (number of strains): spa-t011/agr-II/ST398 (1), spa-t002/agr-II/ST5 (2), spa-t3262/agr-II/ST5 (1), spa-t605/agr-II/ST126 (3), and spa-t878/agr-III/ST2849 (1). S. aureus ST398 strain was methicillin-resistant and showed a multidrug resistance phenotype. Virulence genes tst, etd, sea, sec, seg, sei, sem, sen, seo, and seu, were detected among S. aureus and only ST5 strains showed genes of immune evasion cluster. Thirty-two coagulase-negative Staphylococcus of 12 different species were recovered (number of strains): Staphylococcus equorum (7), Staphylococcus vitulinus (4), Staphylococcus lentus (4), Staphylococcus sciuri (4), Staphylococcus fleurettii (2), Staphylococcus haemolyticus (2), Staphylococcus hominis (2), Staphylococcus saprophyticus (2), Staphylococcus succinus (2), Staphylococcus capitis (1), Staphylococcus cohnii (1), and Staphylococcus epidermidis (1). Five presented a multidrug resistance phenotype. The following resistance and virulence genes were found: mecA, lnu(A), vga(A), tet(K), erm(C), msr(A)/(B), mph(C), tst, and sem. We found that Staphylococcus spp. are normal contaminants of urban wastewater, including different lineages of S. aureus and a high diversity of coagulase-negative species. The presence of multiple resistance and virulence genes, including mecA, in staphylococci of wastewater can be a concern for the public health.

The bacterial diversity in ballast water from five general cargo ships calling at the Port of Houston was determined with ion semiconductor DNA sequencing (Ion Torrent PGM) of PCR amplified 16S rRNA genes. Phylogenetic analysis revealed that the composition of bacteria in ballast water did not resemble that of typical marine habitats or even open ocean waters where BWEs occur. The predominant group of bacteria in ships conducting BWEs was the Roseobacter clade within the Alphaproteobacteria. In contrast, Gammaproteobacteria were predominant in the ship that did not conduct a BWE. All the ships contained human, fish, and terrestrial plant pathogens as well as bacteria indicative of fecal or activated sludge contamination. Most of the 60 pathogens had not been detected in ballast water previously. Among these were the human pathogens Corynebacterium diptheriae and several Legionella species and the fish pathogens Francisella piscicida and Piscirickettsia salmonis.

Marine beaches are important recreational and economic resources in Brazil, but the beaches' water quality is negatively impacted by the discharge of domestic sewage effluent. The occurrence of diarrheagenic Escherichia coli among the E. coli isolated from three Brazilian marine beaches was investigated. Multiplex and single step PCR were used to screen 99 E. coli isolates for ten target toxin genes. Six toxin genes, stx1, eae, estp, esth, astA, and bfpA, were identified in 1% to 35% of the isolates. A quantitative microbial risk assessment (QMRA) of human exposure to diarrheagenic E. coli during marine recreation was carried out. The results indicated that the diarrheagenic E. coli risk is well below the U.S. EPA's recommended daily recreational risk benchmark. However, the overall recreational health risk due to all pathogens in the water could be much higher and exceeded the U.S. EPA's benchmark.

Using an indoor microcosm assay, we analyzed the biodegradation of total petroleum hydrocarbons (TPHs) by autochthonous bacterial populations in mining soil in the presence of a surfactant (Tween 80). The kinetic behavior of TPH biodegradation involved fast and slow stages. Initially, heterotrophic and hydrocarbonoclastic bacteria increased in abundance by an order of magnitude, but both groups decreased to close to their initial population sizes by the end of experiment. The most efficient final biodegradation (61.5 %) was achieved using soil with 0.5 % added surfactant. Polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE) were used to analyze changes in the bacterial community structure. During the fast biodegradation phase, bacterial species richness as indicated by DGGE profiles was reduced after long periods of TPH biodegradation with exposure to Tween 80. The distribution of families was modified, but no particular pattern could be identified. The main bacterial genera were Acinetobacter, Pedomicrobium, Halomonas, Rhizobium, Cryobacterium, Pseudomonas, Lysobacter, Thermomonas, and Stenotrophomonas. Acinetobacter exhibited the highest species richness and was the most abundant and persistent genus, followed by Pedomicrobium and Rhizobium. Decreasing TPH biodegradation can be attributed to a reduction in the microbial population and the disappearance of most of the initial bacterial genera. The correlation between TPH biodegradation and microbial population dynamics helps explain long bioremediation times and can facilitate actions for increasing bioremediation efficiency.

The overapplication of endosulfan on crops has resulted in the widespread contamination of soil. In this study, we examine the potential for bioremediation of the bacteria strain Alcaligenes faecalis JBW4 in degrading endsosulfan in soils. Bacteria were inoculated into sterilized and non-sterilized soils (Argi-Udic Ferrosols and Hapli-Udic Isohumosols) spiked with endosulfan. The results obtained from polymerase chain reaction-denaturing gradient gel electrophoresis indicate that JBW4 colonized Argi-Udic Ferrosols and Hapli-Udic Isohumosols successfully. The degradation efficiencies of α and β isomers of endosulfan by JBW4 were higher in Hapli-Udic Isohumosols than in Argi-Udic Ferrosols, and α and β isomers were degraded by 100.0 and 69.8%, respectively. In addition, detected endosulfan metabolites were either endosulfan ether and endosulfan lactone. Results of the single-cell gel electrophoresis assay showed that the toxicity of endosulfan and its metabolites in Hapli-Udic Isohumosols decreased after 77 days when compared to those in Argi-Udic Ferrosols after degradation by JBW4. Strain JBW4 is an excellent bio-remediator through its ability to degrade endosulfan in contaminated Argi-Udic Ferrosols and Hapli-Udic Isohumosols.

Amaranthus hybridus L. has great potential for use in phytoremediation of soils contaminated with cadmium (Cd). In this study, we found higher absorption of Cd by the roots of A. hybridus than by its other organs. To understand the mechanism of Cd accumulation in A. hybridus roots, a comparative proteomic approach was used to differentiate the two-dimensional electrophoretic profiles of root proteins in Cd-free and Cd-treated plants. Twenty-eight differentially expressed proteins were successfully identified using matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry. Of these, 10 were specifically expressed under Cd stress, and another 11 were upregulated and 7 downregulated by >2.5-fold under Cd stress. We observed increased expression of proteins involved in energy metabolism, protein metabolism, stress and defense, and signal transduction. These changes likely enhanced Cd tolerance and enrichment in A. hybridus. The downregulated proteins were mainly involved in the synthesis of microRNAs, cell walls, and other structural components. These observations were further confirmed by quantitative fluorescence PCR. The resulting differences in protein expression patterns suggest that redirection of root cell metabolism might be an important survival mechanism for A. hybridus under Cd stress.

The aim of this study was the evaluation of the occurrence of pathogenic Campylobacter, Escherichia coli O157:H7, E. coli virulence genes and Salmonella spp. in different wastewater treatment plants (WWTPs) using a method based on an enrichment step and PCR. This method was sensitive enough to detect low levels (∼2 CFU100 ml⁻¹ of raw sewage) of all the investigated pathogens. In the WWTP samples, E. coli O157:H7 DNA and the eae gene were never found, but 33 % of influents and effluents exhibited amplicons corresponding to Shiga-like toxin I. Twenty-five percent of the influent and 8 % of the effluent exhibited the presence of Shiga-like toxin II. Campylobacter jejuni and C. coli DNA were identified in 50 and 25 % of the influents and in 8 and 25 % of the effluents, respectively. Salmonella spp. DNA was present in all the samples. Considering the results obtained, the method tested here offers a reliable and expeditious tool for evaluating the efficiency of the effluent treatment in order to mitigate contamination risk. Influent contamination by Salmonella spp. and Campylobacter spp. provides indirect information about their circulation; moreover, their presence in effluents underlines the role of WWTPs in the contamination of the receiving surface waters, which affects public health directly or indirectly.

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.

Pathogenic Vibrio alginolyticus, a cause of severe infection in shellfish, as well as in humans, has been found at high frequency around all coastal areas of Korea. The aim of this study was to determine the occurrence of V. alginolyticus, to identify the strains isolated from oysters in West Sea, and to investigate their antimicrobial resistance profiles. Biochemical analyses of the 90 initially recovered presumptive V. alginolyticus colonies indicated that 16 isolates were V. alginolyticus. PCR analysis to detect the presence of the gyrB gene confirmed that 15 (93.8 %) of the 16 isolates were V. alginolyticus. These 15 isolates had the following profiles of resistance against 16 antibiotics: all isolates were resistant to ampicillin and vancomycin, and 26.7 % of the isolates exhibited resistance to cephalothin. A large number of isolates showed intermediate resistance to erythromycin (100 %) and rifampin (73.3 %). Five (33.3 %) of the V. alginolyticus isolates demonstrated multiple resistance to at least three antimicrobials.