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ABSTRACT: This study aimed to investigate the chlorination effects on microbial antibiotic resistance in a drinking water treatment plant. Biochemical identification, 16S rRNA gene cloning and metagenomic analysis consistently indicated that Proteobacteria were the main antibiotic resistant bacteria (ARB) dominating in the drinking water and chlorine disinfection greatly affected microbial community structure. After chlorination, higher proportion of the surviving bacteria was resistant to chloramphenicol, trimethoprim and cephalothin. Quantitative real-time PCRs revealed that sulI had the highest abundance among the antibiotic resistance genes (ARGs) detected in the drinking water, followed by tetA and tetG. Chlorination caused enrichment of ampC, aphA2, bla(TEM-1), tetA, tetG, ermA and ermB, but sulI was considerably removed (p < 0.05). Metagenomic analysis confirmed that drinking water chlorination could concentrate various ARGs, as well as of plasmids, insertion sequences and integrons involved in horizontal transfer of the ARGs. Water pipeline transportation tended to reduce the abundance of most ARGs, but various ARB and ARGs were still present in the tap water, which deserves more public health concerns. The results highlighted prevalence of ARB and ARGs in chlorinated drinking water and this study might be technologically useful for detecting the ARGs in water environments.
Water Research 10/2012; · 4.86 Impact Factor
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ABSTRACT: This study aimed to explore the molecular mechanisms behind the stimulation effects of microcystin-LR (a well-known cyanobacterial toxin produced in eutrophic lakes or reservoirs) on cancer cell invasion and matrix metalloproteinases (MMPs) expression. Boyden chamber assay showed that microcystin-LR exposure (>12.5 nM) evidently enhanced the invasion ability of the melanoma cells (MDA-MB-435). Tumor Metastasis PCR Array demonstrated that 24 h microcystin-LR treatment (25 nM) caused overexpression of eight genes involved in tumor metastasis, including MMP-2, MMP-9, and MMP-13. Quantitative real-time PCR, Western blotting and gelatin zymography consistently demonstrated that mRNA and protein levels of MMP-2/-9 were increased in the cells after microcystin-LR exposure (P < 0.05 each). Immunofluorescence assay and electrophoretic mobility shift assay revealed that microcystin-LR could activate nuclear factor kappaB (NF-κB) by accelerating NF-κB translocation into the nucleus and enhancing NF-κB binding ability. Furthermore, addition of NF-κB inhibitor in culture medium could suppress the invasiveness enhancement and MMP-2/-9 overexpression. This study indicates that microcystin-LR can act as a NF-κB activator to promote MMP-2/-9 expression and melanoma cell invasion, which deserves more environmental health concerns.
Environmental Science & Technology 09/2012; 46(20):11319-26. · 4.80 Impact Factor
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ABSTRACT: Mice were fed with source water (SW) and tap water (TW) for 90 d to evaluate hepatotoxicity induced by the drinking water. Histopathologic observation showed no obvious damage to hepatic tissue in the SW and TW groups. However, microarray analysis indicated that the SW and TW exposures affected many metabolic pathways, among which PPAR (peroxisome proliferator-activated receptors) signaling was most susceptible. Immunohistochemical staining demonstrated that both PPAR-α and PPAR-γ were significantly increased in the exposure groups compared to control. Enzyme-linked immunosorbent assay revealed that PPAR-α expression level was increased from 23.37±0.53 ng g(-1) liver weight in control group to 26.60±1.43 ng g(-1) liver weight in SW group and 27.68±1.10 ng g(-1) liver weight in TW group (p<0.05). For PPAR-γ, the expression level was also significantly enhanced from 0.83±0.07 ng g(-1) liver weight in control group to 1.11±0.20 ng g(-1) liver weight in SW group and 1.16±0.07 ng g(-1) liver weight in TW group (p<0.05). The SW and DW posed no obvious hepatotoxicity on mice and PPAR-α/-γ could be used as a novel biomarker to assess public health risk induced by slightly contaminated drinking water.
Chemosphere 03/2012; 88(4):407-12. · 3.21 Impact Factor
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ABSTRACT: The emerging antibiotic resistance genes in the aquatic environment have aroused public concern. As β-lactam is the most widely used group of antibiotics, β-lactam resistance genes were selected to investigate their distribution and diversity in the activated sludge from 15 geographically different sewage treatment plants (STPs) of China, Singapore, USA, and Canada. Specific PCR and quantitative real-time PCR (q-PCR) were used to investigate the occurrence and abundance of nine β-lactam resistance genes. Five genes (OXA-1, OXA-2, OXA-10, ampC, and TEM-1) were detected in most of the sludge collected, while three genes (mecA, CTX-M-1, and SME) were not found in any sludge sample. The total abundances of the six detected β-lactam resistance genes in the 15 STPs varied from 5.34 × 10(1) copies/ng DNA (ampC) to 5.49 × 10(4) copies/ng DNA (OXA-1). Overall, OXA-1 had the highest total concentration, followed by IMP and OXA-10. Noticeably, the abundances of TEM-1 in Chinese STPs were generally higher than those in the STPs of other countries, while the abundances of OXA-2 and IMP in the STPs of North America were much greater than those of East Asia. A total of 78 clones carrying β-lactam resistance genes were randomly selected from six clone libraries for phylogenetic diversity analysis; the similarity of these cloned genes to known β-lactam resistance genes with sequence identities ranged from 96% to 100%. Furthermore, OXA-1, ampC, and IMP were found to be more diverse than the other β-lactam resistance genes.
Applied Microbiology and Biotechnology 12/2011; 95(5):1351-8. · 3.42 Impact Factor
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ABSTRACT: A carcinogenic risk assessment of polycyclic aromatic hydrocarbons (PAHs) in source water and drinking water of China was conducted using probabilistic techniques from a national perspective. The published monitoring data of PAHs were gathered and converted into BaP equivalent (BaP(eq)) concentrations. Based on the transformed data, comprehensive risk assessment was performed by considering different age groups and exposure pathways. Monte Carlo simulation and sensitivity analysis were applied to quantify uncertainties of risk estimation. The risk analysis indicated that, the risk values for children and teens were lower than the accepted value (1.00E-05), indicating no significant carcinogenic risk. The probability of risk values above 1.00E-05 was 5.8% and 6.7% for adults and lifetime groups, respectively. Overall, carcinogenic risks of PAHs in source water and drinking water of China were mostly accepted. However, specific regions, such as Yellow river of Lanzhou reach and Qiantang river should be paid more attention. Notwithstanding the uncertainties inherent in the risk assessment, this study is the first attempt to provide information on carcinogenic risk of PAHs in source water and drinking water of China, and might be useful for potential strategies of carcinogenic risk management and reduction.
Science of The Total Environment 12/2011; 410-411:112-8. · 3.29 Impact Factor
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ABSTRACT: This study assessed the effects of microcystin-LR (MC-LR) exposure on matrix metalloproteinases (MMPs) expression and cancer cell migration. After male mice were orally administered with different concentrations of MC-LR for 270 d, histopathologic observation revealed an obvious hepatic lymphocyte infiltration or fatty degeneration. Immunohistochemical staining and enzyme-linked immunosorbent assay demonstrated that MC-LR treatment (even at 1 nM) caused up-regulated expressions of hepatic MMP-2/-9. Quantitative reverse-transcriptase PCR showed that the exposure to 80 nM MC-LR induced an increase of MMP-2/-9 mRNA levels by 1.0 and 1.9 fold. Breast cancer cells (MDA-MB-435s) were also cultured with MC-LR solutions and a wound healing assay demonstrated that MC-LR posed a time/dose-dependent stimulation effect on migration of the cancer cells. Gelatin electrophoresis and quantitative PCR showed significant increases in cellular MMP-2/-9 expressions after MC-LR exposure. This study indicated that chronic exposure to MC-LR could alter MMP-2/-9 expressions and stimulate cancer cell migration.
Ecotoxicology and Environmental Safety 11/2011; 77:88-93. · 2.29 Impact Factor
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Shi Feng Yun,
Xiao Yun Tian,
Shu Pei Cheng,
Yan Zhang,
Ai Min Li,
Li Bo Zhang, Xu Xiang Zhang,
Li Chen,
Bing Wu,
Lian Qin Guo,
Yun Zhong Shi
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ABSTRACT: The 18 biochemical parameters of serum were measured to analyze the pathogenic risks of the Yangtze River Source of Drinking Water in Nanjing area (YZR-SDW-NJ) on mouse Mus musculus for protection of human health in this research. The male mice Mus musculus were sampled and fed with YZR-SDW-NJ for 90 days then the eighteen serum biochemical levels were measured with Automatic Biochemical Analysis/RerLi 600. And the parameter data were treated by One-Way ANOVA statistic approach. The results showed that five parameter levels for the sample group mice were different from those for the control group significantly (0.01 < P or 0.05 < P). Four 4 of the 5 altered parameter levels were decreased including glutamate pyruvate transaminase 38% lower, glutamine-oxaloacetic transaminase 24% lower, triglyceride 76% lower and cystatin C 73% lower, only creatinine level was 26% higher than that in the control group. The data suggest that YZR-SDW-NJ had toxicity on the mouse and the organic pollutants in YZR-SDW-NJ might lead to liver, kidney, cardiovascular and metabolic pathogenic risks on the human beings. The results might be cited as evidence to control pollutants in the source water for the protection of NJ people's health.
Ecotoxicology 07/2011; 20(5):1078-82. · 2.36 Impact Factor
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ABSTRACT: Water from Taihu Lake (China) is used as a drinking source. The frequency of contamination in Taihu Lake has increased over the past decade and the bloom-forming cyanobacteria are the dominant species during eutrophication. Cyanobacteria can produce various harmful secondary substances including microcystins capable of endangering human health and ecological safety. This study investigated toxicity of ingested Taihu Lake water on mice via hepatic histopathology and matrix metalloproteinase (MMP) expression. Water was sampled from four Taihu Lake locations, Meiliang Bay 2 group (M2), Meiliang Bay 1 group (M1), Lake Center (H) and Xukou Bay (X), along a gradient of decreasing degree of eutrophication. The experimental design consists of five groups of male mice (Mus musculus, ICR): one control and four groups ingesting water from the four sampling sites for 90 days. Compared to control, M1 and M2 mice showed hepatic histopathological changes including swollen, vacuolar degeneration or inflammatory. Immunohistochemical staining demonstrated a higher expression of MMP-2 proteins in M2 group and a lower expression of MMP-9 in M1. Enzyme-linked immunosorbent assay indicated that MMP-9 concentration was significantly reduced from 0.55 to 0.28 ng/g liver weight in M2 (p < 0.05). Real time PCR revealed a down-regulation of MMP-9 mRNA by 2.2 fold in M1 and an up-regulation of MMP-2 mRNA by 1.73 fold in H. Using this mouse model as a gauge of water toxicity, our results revealed that potential health risks induced by Taihu Lake water might arise from the use of this source water by local resident.
Ecotoxicology 03/2011; 20(5):1047-56. · 2.36 Impact Factor
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ABSTRACT: Activated sludge was sampled from 15 sewage treatment plants (STPs) across China and other global locations to investigate the occurrence, abundance and diversity of tetracycline resistance genes (tet) in the STPs. Occurrence and abundance of 14 tet genes were determined using polymerase chain reaction (PCR) and quantitative real time PCR. Six genes (tet(A), tet(C), tet(G), tet(M), tet(S), and tet(X)) were detected in all the STPs, while no sludge sample contained tet(Q). Total concentration of the 14 genes was significantly different among the STPs and average tet abundance of the STPs varied greatly among the tet types (p<0.05). Tet(G) had the highest concentration in the STPs, followed by tet(C), tet(A) and tet(S). Phylogenetic diversity of the genes was investigated using DNA cloning. BLAST analysis showed that all of the 450 cloned sequences matched known tet genes, except for tet(G). The 56 tet(G) clones were grouped into 14 genotypes, among which type G24 had an identical sequence to tet(G) carried by Salmonella enterica or Acinetobacter baumannii, while the other sequences had low similarity to the known genes in GenBank. The results of this study might be useful to understand the diversity of these resistance genes in STPs.
Environmental Science & Technology 03/2011; 45(7):2598-604. · 4.80 Impact Factor
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ABSTRACT: Cytochrome P450 2E1 (CYP2E1) is a cytochrome P450 enzyme involved in styrene metabolism. This study compared the binding affinities between styrene and 11 mammalian CYP2E1 systems using bioinformatics methods. Firstly, amino acid sequences of CYP2E1s were obtained from the Swiss-Prot database. Then, taking the crystal structure of human CYP2E1 as a template, 3D models of the CYP2E1s of other mammals were constructed using the SWISS-MODEL program. Finally, the generated homology models were applied to calculate their docking capacities against styrene and polystyrene using the Surflex-Dock program, which could automatically dock ligands into a receptor's ligand binding site using a protomol based approach and assess the affinity by an empirically derived scoring function. Docking experiments showed that the studied mammalian CYP2E1s had high binding affinities with styrene. For polystyrene, the dimmer of styrene has high binding affinities with CYP2E1s, however, trimer and other high polymers were found hard to be docked into the CYP2E1s. The results of this study indicated that bioinformatics approaches might be useful tools to predict styrene and polystyrene affinities with mammalian CYP2E1s.
Ecotoxicology 03/2011; 20(5):1041-6. · 2.36 Impact Factor
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ABSTRACT: A probability risk assessment of anthracene, benzo(a)pyrene, chrysene, fluorene, phenanthrene and pyrene was carried out to examine the ecological risk of these six polycyclic aromatic hydrocarbons (PAHs) in aquatic ecosystems in China. The literature on PAH concentrations in surface water in China was collected to evaluate the environmental exposure concentrations (EEC). The 10th percentile of predicted no observed effect concentration (PNEC(10%)) of PAHs, calculated according to the data from the USEPA AQUIRE database and regulatory reviews, was applied as the toxicity assessment endpoint. The ratio of EEC and PNEC(10%), expressed as a risk quotient (RQ), was used to characterize the risk value. Bootstrapping method and Monte Carlo simulation were utilized to calculate the distribution of EEC, PNEC(10%), RQ and associated uncertainties. Risk assessment showed that reliable maximum RQs of anthracene, benzo(a)pyrene, chrysene, fluorene and phenanthrene were in the range of 0.064-0.755, lower than the acceptable value of 1. However, the reliable maximum RQ of pyrene was 1.39, indicating its potential ecological risk. Notwithstanding the uncertainty, these results suggest that the aquatic ecosystems with high PAH concentrations might pose potential ecological risks, and concerted efforts are required to ensure that surface water is protected.
Ecotoxicology 03/2011; 20(5):1124-30. · 2.36 Impact Factor
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ABSTRACT: Integrons are elements that encode a site-specific recombination system that recognizes and captures mobile gene cassettes and are closely related to multiple resistances of environmental microorganisms. This study was undertaken to determine the efficiency of an activated sludge process to remove integrons. The prevalence and characteristics of class 1 integrons were investigated for bacterial species isolated from the activated sludge of Nanjing Jiangxinzhou sewage treatment plant (STP, China). A total of 189 bacterial strains were isolated from influent water, activated sludge and effluent water, and PCR-RFLP (Polymerase chain reaction--restriction fragment length polymorphism) of 16S rRNA gene showed that the isolated bacteria were Escherichia coli, Aeromonas veronii, Klebsiella spp., Aeromonas salmonicida and Aeromonas media. PCRs showed that 57 isolates contained class 1 integronase gene intI1. The integron detection frequency in the isolated strains was 20.4% for influent, 30.9% for activated sludge and 38.9% for effluent. Quantitative real-time PCR assay showed that the abundance of integrons in effluent was higher than that in influent. This study indicates that class 1 integrons are wide-spread in STPs which might be involved in multiple resistances in the activated sludge characterized by high biomass and biodiversity.
Ecotoxicology 03/2011; 20(5):968-73. · 2.36 Impact Factor
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ABSTRACT: The overuse or misuse of antibiotics has accelerated antibiotic resistance, creating a major challenge for the public health in the world. Sewage treatment plants (STPs) are considered as important reservoirs for antibiotic resistance genes (ARGs) and activated sludge characterized with high microbial density and diversity facilitates ARG horizontal gene transfer (HGT) via mobile genetic elements (MGEs). However, little is known regarding the pool of ARGs and MGEs in sludge microbiome. In this study, the transposon aided capture (TRACA) system was employed to isolate novel plasmids from activated sludge of one STP in Hong Kong, China. We also used Illumina Hiseq 2000 high-throughput sequencing and metagenomics analysis to investigate the plasmid metagenome. Two novel plasmids were acquired from the sludge microbiome by using TRACA system and one novel plasmid was identified through metagenomics analysis. Our results revealed high levels of various ARGs as well as MGEs for HGT, including integrons, transposons and plasmids. The application of the TRACA system to isolate novel plasmids from the environmental metagenome, coupled with subsequent high-throughput sequencing and metagenomic analysis, highlighted the prevalence of ARGs and MGEs in microbial community of STPs.
PLoS ONE 01/2011; 6(10):e26041. · 4.09 Impact Factor
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ABSTRACT: In order to investigate the potential effects of microcystin-LR (MC-LR) on the expression of metalloproteinases (MMPs), Mice were orally administered with MC-LR in drinking water (0, 1, 40 and 80 μg/L) for 180 d, and hepatic MMP-2/-9 expression was evaluated at the levels of enzyme activity, protein level and mRNA expression. Histopathologic observation showed the obvious hepatic lymphocyte infiltration and fatty degeneration in the mice exposed to 40 and 80 μg/L MC-LR. Immunohistochemical staining and enzyme-linked immunosorbent assay (ELISA) revealed that excess MMP-2/-9 proteins were produced in livers of the mice exposed to MC-LR at the higher concentrations. Hepatic MMP-9 level was elevated from 0.6 ng/g liver weight in control to 1.4 ng/g liver weight in 80-μg/L group, but a slight increase was found for MMP-2 level. Real time PCR showed that MMP-2/-9 mRNA expression was up-regulated by 6.9 fold and 5.0 fold after 80-μg/L-MC treatment, respectively. MMP-2/-9 expression showed a good dose-dependent manner at both protein and mRNA levels. ELISA demonstrated that MC-LR stimulated phosphorylation of mitogen-activated protein kinases, a potential signal transduction pathway of the MMP-2/-9 expression alteration. This study revealed a significant alteration in hepatic MMP-2/-9 expression induced by MC-LR, which might be involved in cell invasion and metastasis.
Toxicology Letters 10/2010; 199(3):377-82. · 3.23 Impact Factor
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ABSTRACT: DNA micorarray was used to analyze hepatic transcriptional profile of male mice (Mus musculus) after 35-d intragastric perfusion treatment with purified terephthalic acid (PTA) manufacturing wastewater. Haematological analysis demonstrated that the levels of glutamyl transferase and lactate dehydrogenase in serum were significantly decreased, and DNA microarray showed that a total of 306 genes were differentially expressed in PTA wastewater-treated mice. According to Kyoto encyclopedia of genes and genomes pathway database, the differentially expressed genes were mainly grouped to metabolic pathways (58 genes) and biological processes (101 genes). PTA wastewater had significant impacts upon metabolisms of lipid, carbohydrate, amino acid, vitamin and nucleotide. Several signal transduction pathways are most susceptible to PTA wastewater, including mitogen-activated protein kinases, Janus kinase/signal transducers and activators of transcription and calcium signaling pathways. Potential public health problems may arise from the discharge of PTA wastewater into the environment.
Journal of hazardous materials 09/2010; 181(1-3):1121-6. · 4.14 Impact Factor
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ABSTRACT: Homology modeling and molecular docking were used to in silico analyze the interactions between six endocrine disrupting chemicals (EDCs) and 11 androgen receptors (ARs) of different vertebrate species. The MODELLER 9V7 program was employed to construct the homology models of AR ligand binding domains (LBDs) from birds, amphibians, bony fishes and cartilaginous fishes. The Surflex-Dock program was applied to calculate and analyze the binding affinities between the six EDCs and AR LBDs. The docking experiment showed that AR LBDs had high affinities with nonyl phenol (NP) and butyl benzyl phthalate (BBP), but low affinities with the 2,2',4,4',5,5'-hexabromodiphenyl ether (BDE153). The results of cluster analysis suggested that predicted binding affinities were species-specific, which was consistent with the phylogenetic analysis of AR LBDs. The difference of binding affinities could be mainly due to the different hydrogen bonds and the orientation of ligands in the binding pockets. Our results suggest that integrated methods of phylogenetic analysis, homology modeling and molecular docking might be a potential tool to predict the different interactions between contaminants and associated receptors in different trophic levels.
Chemosphere 07/2010; 80(5):535-41. · 3.21 Impact Factor
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ABSTRACT: Six biofilters were used for advanced treatment of Yangtze River source water to investigate the effects of hydraulic loading rate (HLR) on pollutant removal and microbial community. HLR was found to exert significant influences on the removal efficiency of the conventional pollutants and 24 detectable semivolatile organic compounds (SVOCs). More than 85% of chemical oxygen demand and assimilable organic carbon was removed at the optimal HLR of 3.0 m h(-1). With the increase of HLR, SVOC removal showed a decreasing trend. Di-n-butyl phthalate and bis(2-ethylhexyl)phthalate, two main SVOCs in the source water, had the highest removals of 71.2% and 84.4%, respectively. Nearly 65% of 2,6-dinitrotoluene and 80% of isophorone were removed at the lowest HLR. Phylogenetic analysis showed that Escherichia coli, Shigella sp., E. fergusonii and Firmicutes bacteria predominated in the bioreactors. The dominance of E. coli in the low-HLR biofilters might contribute greatly to the high SVOC removal.
Journal of hazardous materials 02/2010; 178(1-3):652-7. · 4.14 Impact Factor
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ABSTRACT: Reproductive toxicity of purified terephthalic acid (PTA) manufacturing wastewater on the male mice (Mus musculus) was investigated after 35-day intragastric perfusion treatment with the wastewater. Fluorescein diacetate and propidium iodide staining, and flow cytometry were used to assess the toxicity of PTA wastewater on spermatogenic cells. PTA wastewater induced significant variations in the relative percentages of immature haploid, diploid, tetraploid and S-phase spermatogonia. Percentage of viable spermatogenic cells was reduced from 93.1+/-2.3 in control group to 90.4+/-1.9 in the wastewater-treated group. Testicular histopathology revealed expansion of interstitial space and reduction in the number and size of Leydig cells induced by the wastewater, which was further certified by the decrease (10.6%) in relative testes weight and the increase (101.3%) in sperm shape abnormality in the wastewater-treated group. In this study, PTA wastewater was found to have reproductive toxicity on male mice, and public health problems may potentially arise from the discharge of the wastewater into the environment.
Journal of hazardous materials 11/2009; 176(1-3):300-5. · 4.14 Impact Factor
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ABSTRACT: Class 1 integrons and gene cassettes containing antibiotic resistance genes (ARGs) in five different sewage treatment plants (STPs) were characterized and quantified using polymerase chain reaction (PCR), sequencing, and quantitative real-time PCR (qRT-PCR) in this study. Class 1 integronase gene (intI1) was found commonly occurring in all of activated sludge samples from the five STPs, as well as in influent and effluent of two STPs at Hong Kong. One hundred and nine lactose-fermenting Enterobacteriaceae (LFE) strains were isolated from activated sludge of Shatin STP. Among them, 36 strains (33.0%) were found to carry class 1 integrons. PCR assays showed that 11 of the 36 intI1-carrying isolates harbored a common type of gene cassette array of about 1,600 bps, as well as the static genes (sulI and qacEDelta1) on class 1 integrons. This gene cassette array was found phylogenetically close to antibiotic resistance genes dfr17 and aadA5, encoding dihydrofolate reductase conferring resistance to trimethoprim and adenylyltransferase conferring resistance to spectinomycin/streptomycin, respectively. Antimicrobial susceptibility analysis demonstrated that all the 11 LFEs carrying gene cassette were multi-resistant, especially having common resistance to trimethoprim and streptomycin. qRT-PCR assay showed that genes copies of both class 1 integron and the gene cassette varied significantly among the activated sludge sampled from different STPs, at different time points or different treatment steps. More than 90% of class 1 integrons and the gene cassette were removed by activated sludge processes in two STPs, while the disinfection process removed 94% integron and 77% gene cassette in one STP.
Applied Microbiology and Biotechnology 03/2009; 82(6):1169-77. · 3.42 Impact Factor
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ABSTRACT: The use of antibiotics may accelerate the development of antibiotic resistance genes (ARGs) and bacteria which shade health risks to humans and animals. The emerging of ARGs in the water environment is becoming an increasing worldwide concern. Hundreds of various ARGs encoding resistance to a broad range of antibiotics have been found in microorganisms distributed not only in hospital wastewaters and animal production wastewaters, but also in sewage, wastewater treatment plants, surface water, groundwater, and even in drinking water. This review summarizes recently published information on the types, distributions, and horizontal transfer of ARGs in various aquatic environments, as well as the molecular methods used to detect environmental ARGs, including specific and multiplex PCR (polymerase chain reaction), real-time PCR, DNA sequencing, and hybridization based techniques.
Applied Microbiology and Biotechnology 02/2009; 82(3):397-414. · 3.42 Impact Factor
