Incidence of metal and antibiotic resistance in Pseudomonas spp. from the river water, agricultural soil irrigated with wastewater and groundwater

A total of 144 isolates of Pseudomonas spp. (48 each from the Yamuna River water, wastewater irrigated soil and groundwater irrigated soil) were tested for their resistance against certain heavy metals and antibiotics. Minimum inhibitory concentrations (MICs) of Hg^2?+?, Cd^2?+?, Cu^2?+?, Zn^2?+?, Ni^2?+?, Pb^2?+?, Cr^3?+? and Cr^6?+? for each isolate were also determined. A maximum MIC of 200 ??g/ml for mercury and 3,200 ??g/ml for other metals were observed. The incidences of metal resistance and MICs of metals for Pseudomonas isolates from the Yamuna water and wastewater irrigated soil were significantly different to those of groundwater irrigated soil. A high level of resistance against tetracycline and polymyxin B (81.2%) was observed in river water isolates. However, 87.5% of Pseudomonas isolates from soil irrigated with wastewater showed resistance to sulphadiazine, whereas 79.1% were resistant to both ampicillin and erythromycin. Isolates from soil irrigated with groundwater exhibited less resistance towards heavy metals and antibiotics as compared to those of river water and wastewater irrigated soil. Majority of the Pseudomonas isolates from water and soil exhibited resistance to multiple metals and antibiotics. Resistance was transferable to recipient Escherichia coli AB2200 strains by conjugation. Plasmids were cured with the curing agent ethidium bromide and acridine orange at sub-MIC concentration.     

Assessment of physicochemical parameters and prevalence of virulent and multiple-antibiotic-resistant <Emphasis Type="Italic">Escherichia coli</Emphasis> in treated effluent of two wastewater treatment plants and receiving aquatic <Emphasis Type="Italic">milieu</Emphasis> in Durban,South Africa

The poor operational status of some wastewater treatment plants often result in the discharge of inadequately treated effluent into receiving surface waters. This is of significant public health concern as there are many informal settlement dwellers (ISDs) that rely on these surface waters for their domestic use. This study investigated the treatment efficiency of two independent wastewater treatment plants (WWTPs) in Durban, South Africa and determined the impact of treated effluent discharge on the physicochemical and microbial quality of the receiving water bodies over a 6-month period. Presumptive Escherichia coli isolates were identified using biochemical tests and detection of the mdh gene via PCR. Six major virulence genes namely eae, hly, fliC, stx1, stx2, and rfbE were also detected via PCR while antibiotic resistance profiles of the isolates were determined using Kirby-Bauer disc diffusion assay. The physicochemical parameters of the wastewater samples ranged variously between 9 and 313.33 mg/L, 1.52 and 76.43 NTUs, and 6.30 and 7.87 for COD, turbidity, and pH respectively, while the E. coli counts ranged between 0 and 31.2?×?10³ CFU/ml. Of the 200 selected E. coli isolates, the hly gene was found in 28 %, fliC in 20 %, stx2 in 17 %, eae in 14 %, with stx1 and rfbE in only 4 % of the isolates. Notable resistance was observed toward trimethoprim (97 %), tetracycline (56 %), and ampicillin (52.5 %). These results further highlight the poor operational status of these WWTPs and outline the need for improved water quality monitoring and enforcement of stringent guidelines.     

Speciation and antimicrobial resistance of Enterococci isolated from recreational beaches in Malaysia

We report the first study on the occurrence of antibiotic-resistant enterococci in coastal bathing waters in Malaysia. One hundred and sixty-five enterococci isolates recovered from two popular recreational beaches in Malaysia were speciated and screened for antibiotic resistance to a total of eight antibiotics. Prevalence of Enterococcus faecalis and Enterococcus faecium was highest in both beaches. E. faecalis/E. faecium ratio was 0.384:1 and 0.375:1, respectively, for isolates from Port Dickson (PD) and Bagan Lalang (BL). Analysis of Fisher’s exact test showed that association of prevalence of E. faecalis and E. faecium with considered locations was not statistically significant (p?<?0.05). Chi-square test revealed significant differences (χ ²?=?82.630, df?=?20, p?<?0.001) in the frequency of occurrence of enterococci isolates from the considered sites. Resistance was highest to nalidixic acid (94.84 %) and least for chloramphenicol (8.38 %). One-way ANOVA using Tukey–Kramer multiple comparison test showed that resistance to ampicillin was higher in PD beach isolates than BL isolates and the difference was extremely statistically significant (p?<?0.0001). Frequency of occurrence of multiple antibiotic resistance (MAR) isolates were higher for PD beach water (64.29 %) as compared to BL beach water (13.51 %), while MAR indices ranged between 0.198 and 0.48. The results suggest that samples from Port Dickson may contain MAR bacteria and that this could be due to high-risk faecal contamination from sewage discharge pipes that drain into the sea water.     

A pilot study on water pollution and characterization of multidrug-resistant superbugs from Byramangala tank, Ramanagara district, Karnataka, India

Urbanization and industrialization has increased the strength and qualities of municipal sewage in Bangalore, India. The disposal of sewage into natural water bodies became a serious issue. Byramangala reservoir is one such habitat enormously polluted in South India. The water samples were collected from four hotspots of Byramangala tank in 3 months. The biochemical oxygen demand (BOD) and bacterial counts were determined. The fecal coliforms were identified by morphological, physiological, and biochemical studies. The antibiotics sensitivity profiling of isolated bacteria were further carried out. We have noticed that a high content of BOD in the tank in all the 3 months. The total and fecal counts were found to be varied from 1.6?×?10⁶ to 8.2?×?10⁶?colony forming unit/ml and >5,500/100 ml, respectively. The variations in BOD and total count were found to be statistically significant at p?>?0.05. Many pathogenic bacteria were characterized and most of them were found to be multidrug resistant. Salmonella showed resistance to cefoperazone, cefotaxime, cefixime, moxifloxacin, piperacillin/tazobactam, co-trimoxazole, levofloxacin, trimethoprim, and ceftazidime. Escherichia coli showed resistance to chloramphenicol, trimethoprim, co-trimoxazole, rifampicin, and nitrofurantoin while Enterobacter showed resistant to ampicillin, cefepime, ceftazidime, cefoperazone, and cefotaxime. Klebsiella and Shigella exhibited multiple drug resistance to conventional antibiotics. Staphylococcus showed resistance to vancomycin, methicillin, oxacillin, and tetracycline. Furthermore, Salmonella and Klebsiella are on the verge of acquiring resistance to even the strongest carbapenems-imipenem and entrapenem. Present study revealed that Byramanagala tank has become a cesspool of multidrug-resistant “superbugs” and will be major health concern in South Bangalore, India.     

Bacteria with dual resistance to elevated concentrations of heavy metals and antibiotics in Nigerian contaminated systems

Samples of soil, water, and sediments from industrial estates in Lagos were collected and analyzed for heavy metals and physicochemical composition. Bacteria that are resistant to elevated concentrations of metals (Cd^2?+?, Co^2?+?, Ni^2?+?, Cr^6?+?, and Hg^2?+?) were isolated from the samples, and they were further screened for antibiotic sensitivity. The minimum tolerance concentrations (MTCs) of the isolates with dual resistance to the metals were determined. The physicochemistry of all the samples indicated were heavily polluted. Twenty-two of the 270 bacterial strains isolated showed dual resistances to antibiotics and heavy metals. The MTCs of isolates to the metals were 14 mM for Cd^2?+?, 15 mM for Co^2?+? and Ni^2?+?, 17 mM for Cr^6?+?, and 10 mM for Hg^2?+?. Five strains (Pseudomonas aeruginosa, Actinomyces turicensis, Acinetobacter junni, Nocardia sp., and Micrococcus sp.) resisted all the 18 antibiotics tested. Whereas Rhodococcus sp. and Micrococcus sp. resisted 15 mM Ni^2?+?, P. aeruginosa resisted 10 mM Co^2?+?. To our knowledge, there has not been any report of bacterial strains resisting such high doses of metals coupled with wide range of antibiotics. Therefore, dual expressions of antibiotics and heavy-metal resistance make the isolates, potential seeds for decommissioning of sites polluted with industrial effluents rich in heavy metals, since the bacteria will be able to withstand in situ antibiosis that may prevail in such ecosystems.     

The occurrence of Shiga toxin-producing Escherichia coli in bathing water of the Sierra de la Ventana region, Buenos Aires Province, Argentina

The region of Sierra de la Ventana is located in the southwest of Buenos Aires Province, Argentina. Traditionally, this area has been devoted to livestock and agriculture, but tourism has had a significant development in recent years. In the region, there are many rivers and streams that are used for swimming and bathing. A survey of the occurrence of Shiga toxin-producing Escherichia coli (STEC) in these waters was conducted, and the microbiological quality of rivers and streams was investigated. No E. coli O157 was recovered by immunomagnetic separation. Nevertheless, the Shiga toxin gene, exclusively stx2 genotype, was detected in four non-O157 E. coli strains. Two STEC strains carried eae factor, but none of them harbored the EHEC-hlyA gene. Three of the STEC isolates belonged to samples obtained in the warm months, and one to the winter sampling. In the sample sites where STEC strains were isolated the counts of E. coli/100 ml exceeded or were close to the limit recommended by the United States Environmental Protection Agency for bathing water. The relationship observed between the rainy season and E. coli counts suggests that among the main causes for the hygienic indicator increase is the runoff of manure deposited on soils that may also induce the entrance of pathogens into the aquatic environment. This research, the first reporting STEC isolation from recreational waters in this area, revealed that streams and rivers from a beef-producing area of Argentina are a reservoir of STEC strains.     

A geographical approach to tracking Escherichia coli and other water quality constituents in a Texas coastal plains watershed

Diffuse sources of surface water pathogens and nutrients can be difficult to isolate in larger river basins. This study used a geographical or nested approach to isolate diffuse sources of Escherichia coli and other water quality constituents in a 145.7-km² river basin in south central Texas, USA. Average numbers of E. coli ranged from 49 to 64,000 colony forming units (CFU) per 100 mL depending upon season and stream flow over the 1-year sampling period. Nitrate-N concentrations ranged from 48 to 14,041 μg?L^?1 and orthophosphate-P from 27 to 2,721 μg?L^?1. High concentrations of nitrate-N, dissolved organic nitrogen, and orthophosphate-P were observed downstream of waste water treatment plants but E. coli values were higher in a watershed draining an older part of the city. Total urban land use explained between 56 and 72 % of the variance in mean annual E. coli values (p?<?0.05) in nine hydrologically disconnected creeks. Of the types of urban land use, commercial land use explained most of the variance in E. coli values in the fall and winter. Surface water sodium, alkalinity, and potassium concentrations in surface water were best described by the proportion of commercial land use in the watershed. Based on our nested approach in examining surface water, city officials are able to direct funding to specific areas of the basin in order to mitigate high surface water E. coli numbers and nutrient concentrations.     

Prevalence of antibiotic-resistant bacteria in three different aquatic environments over three seasons

The objective of this study was to evaluate the impact of urbanization and seasonal changes on the prevalence of antibiotic-resistant bacteria in different aqueous environments. To this end, bacteria were isolated from three different water sources: the River Hooghly in Kolkata, River Kangsabati and groundwater from Kharagpur, West Bengal over three seasons: post-monsoon, winter and summer in 2012–2013. A total of 163 Gram-negative bacteria were isolated from the River Hooghly (n?=?138), River Kangsabati (n?=?13) and groundwater (n?=?12). Antibiotic susceptibility testing was done using 12 antibiotic discs. The percentages of multiple antibiotic-resistant (MAR) bacteria at the three sampling locations were found to be 71.01 % (98/138) for River Hooghly, 15.38 % (2/13) for River Kangsabati and 8.33 % (1/12) for groundwater. Prevalence of MAR bacteria with respect to the three seasons were the following: 73.58 % in post-monsoon, 59.26 % in winter and 53.57 % in summer. Antibiotic resistance index (ARI) was calculated for each location and each season. In general, ARI values for all the River Hooghly samples were >0.2 while those for the River Kangsabati and groundwater in Kharagpur were always <0.2 indicating greater exposure to antibiotics and subsequent resistance in bacteria from the River Hooghly compared to the other two locations. In addition, percentage of MAR and ARI values followed the trend: post-monsoon?>?winter?>?summer. This may be due to the additional terrestrial resistants that get swept along with surface runoff during the monsoons.     

Assessment of animal impacts on bacterial water quality in a South Carolina, USA tidal creek system

Fecal pollution may adversely impact water quality in coastal ecosystems. The goal of this study was to determine whether cattle were a source of fecal pollution in a South Carolina watershed. Surface water samples were collected in June 2002 and February through March 2003 in closed shellfish harvesting waters of Toogoodoo Creek in Charleston County, SC. Fecal coliform concentrations in 70 % of the water samples taken for this study exceeded shellfish harvesting water standards. Ribotyping was performed in order to identify animal sources contributing to elevated fecal coliform levels. Escherichia coli isolates (n?=?253) from surface water samples were ribotyped and compared to a ribotype library developed from known sources of fecal material. Ribotypes from water samples that matched library ribotypes with 90 % maximum similarity or better were assigned to that source. Less than half of the unknown isolates (38 %) matched with library isolates. About half (53 %) of the matched ribotypes were assigned to cattle isolates and 43 % to raccoon. Ribotyping almost exclusively identified animal sources. While these results indicate that runoff from cattle farms was a likely source of fecal pollution in the watershed, wildlife also contributed. Given the small size of the library, ribotyping was moderately useful for determining the impact of adjacent cattle farms on Toogoodoo Creek. Increasing the number and diversity of the wildlife sources from the area would likely increase the usefulness of the method.     

Antibiotic resistance profile of bacterial isolates from animal farming aquatic environments and meats in a peri-urban community in Daejeon, Korea

The increasing usage of antibiotics in the animal farming industry is an emerging worldwide problem contributing to the development of antibiotic resistance. The purpose of this work was to investigate the prevalence and antibiotic resistance profile of bacterial isolates collected from animal farming aquatic environments and meats in a peri-urban community in Daejeon, Korea. In an antibacterial susceptibility test, the bacterial isolates showed a high incidence of resistance (～26.04%) to cefazolin, tetracycline, gentamycin, norfloxacin, erythromycin and vancomycin. The results from a test for multiple antibiotic resistance indicated that the isolates were displaying an approximately 5-fold increase in the incidence of multiple antibiotic resistance to combinations of two different antibiotics compared to combinations of three or more antibiotics. Most of the isolates showed multi-antibiotic resistance, and the resistance patterns were similar among the sampling groups. Sequencing data analysis of 16S rRNA showed that most of the resistant isolates appeared to be dominated by the classes Betaproteobacteria and Gammaproteobacteria, including the genera Delftia, Burkholderia, Escherichia, Enterobacter, Acinetobacter, Shigella and Pseudomonas.     

Monitoring of multiple drug-resistant pathogens in a selected stretch of Bay of Bengal,India

The present work aims at identification of multiple drug-resistant pathogenic bacteria in a selected stretch, namely, Puri on the Bay of Bengal, India. Six stations at the coast of Puri were selected and samples of water and sediment were collected during the winter of 2008 and 2009 for this study. Thirty-eight pathogenic bacteria were isolated and identified from both the water and the sediment of 6 fixed stations (PU-1a, PU-1b, PU-2, PU-3, PU-4, and PU-5). The identified pathogens were Escherichia coli, Vibrio cholerae, Vibrio parahaemolyticus, Klebsiella pneumoniae, Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aeruginosa, and Proteus mirabilis. Antibiotic sensitivity of the isolated bacteria was studied by using 12 selected antibiotics, commonly used for the medication of human beings and animals. The isolated pathogens from both the water and the sediment samples showed lowest resistance to chloramphenicol (C-30 μg) where as the pathogens showed highest level of resistance to ampicillin (10-μg) among the antibiotics used for the study. Among the isolated pathogens E. faecalis (PU-1a), P. aeruginosa (PU-2 and PU-3), E. coli (PU-3 and PU-4), and K. pneumonia (PU-4) showed resistance to more than four antibiotics. Out of the isolated species, 57.8% pathogens were multi-drug resistant. Antibiotic resistance indexes of all the stations were calculated and found to be in the range of 0.066 to 0.083.     

Aeromonas and Pseudomonas: antibiotic and heavy metal resistance species from Iskenderun Bay, Turkey (northeast Mediterranean Sea)

We studied the susceptibility patterns to 15 different antibiotics and six heavy metals in Aeromonas spp. and Pseudomonas spp. isolated from Iskenderun Bay, Turkey (northeast Mediterranean Sea). A high percentage of Aeromonas isolates showed resistance to cefazolin (66.6%) and trimethoprim–sulphamethoxazole (66.6%). Amongst the Pseudomonas isolates, there was a high incidence of resistance to nitrofurantoin (86.2%), cefazolin (84.8%) and cefuroxime (71.7%). Most isolates showed tolerance to different concentrations of heavy metals, and minimal inhibition concentrations ranged from 25 to >3,200 μg/ml. The Aeromonas spp. and Pseudomonas spp. showed high resistance to copper of 98.3% and 75.4%, respectively, and low resistance to lead of 1.7% and 7.2%, respectively. Our results show that antibiotic and heavy metal resistant Aeromonas spp. and Pseudomonas spp. were widespread in Iskenderun Bay in 2007 and 2008. The increasing presence of antibiotic and heavy metal resistant Aeromonas spp. and Pseudomonas spp. may become a potential human health hazard.     

Development of a PCR protocol for the detection of Escherichia coli O157:H7 and Salmonella spp. in surface water

Escherichia coli O157:H7 and Salmonella are pathogenic microorganisms that can cause severe gastrointestinal illness in humans. These pathogens may be transmitted in a variety of ways, including food and water. The presence of Salmonella and E. coli O157:H7 in surface waters constitutes a potential threat to human health when used for either drinking or recreation. As with most waterborne pathogens, Salmonella and E. coli O157:H7 are difficult to detect and enumerate with accuracy in surface waters due to methodological limitations. The aim of this study was to develop a protocol for the detection of Salmonella spp., E. coli O157:H7 and E. coli virulence genes (stx ₁, stx ₂ and eae) in water using a single enrichment step and PCR. In spiked water samples, PCR results showed high sensitivity (<3 CFU/L) for both microorganisms. The protocol developed in this study has been applied in different surface waters in association with microbiological and physical analysis. The frequency of PCR positive samples was 33% for Salmonella and 2% for E. coli O157:H7 producing intimin (eae) and Shiga-like toxin I (stx ₁). Moreover, the finding of amplicons corresponding to eae and stx ₁ genes in the absence of E. coli O157:H7 suggested the possible presence of other pathogenic bacteria that carry these genes (e.g. EHEC, Shigella strains). The results obtained showed that the developed protocol could be applied as a routine analysis of surface water for the evaluation of microbiological risks.     

High-level aminoglycoside resistance and virulence characteristics among Enterococci isolated from recreational beaches in Malaysia

We report the first study on the occurrence of high-level aminoglycoside-resistant (HLAR) Enterococci in coastal bathing waters and beach sand in Malaysia. None of the encountered isolates were resistant to high levels of gentamicin (500 μg/mL). However, high-level resistance to kanamycin (2,000 μg/mL) was observed in 14.2 % of tested isolates, the highest proportions observed being among beach sand isolates. High-level resistance to kanamycin was higher among Enterococcus faecalis and Enterococcus faecium than Enterococcus spp. Chi-square analysis showed no significant association between responses to tested antibiotics and the species allocation or source of isolation of all tested Enterococci. The species classification of encountered Enterococci resistance to vancomycin was highest among Enterococcus spp. (5.89 %) followed by E. faecium (4.785) and least among E. faecalis. A total of 160 isolates were also tested for virulence characteristics. On the whole, caseinase production was profoundly highest (15.01 %) while the least prevalent virulence characteristic observed among tested beach Enterococci was haemolysis of rabbit blood (3.65 %). A strong association was observed between the source of isolation and responses for each of caseinase (C?=?0.47, V?=?0.53) and slime (C?=?0.50, V?=?0.58) assays. Analysis of obtained spearman’s coefficient showed a strong correlation between caseinase and each of the slime production (p?=?0.04), gelatinase (p?=?0.0035) and haemolytic activity on horse blood (p?=?0.004), respectively. Suggestively, these are the main virulent characteristics of the studied beach Enterococci. Our findings suggest that recreational beaches may contribute to the dissemination of Enterococci with HLAR and virulence characteristics.     

The prevalence of bacterial resistance in clinical, food, water and some environmental samples in Southwest Nigeria

The resistance pattern and mechanisms of bacterial isolates obtained from clinical origin, soil, industrial effluent, orange juice products and drinking water were studied using commonly used antibiotics. The microbial load of the water samples, industrial effluent and orange juice products were 1.0 × 10¹–2.25 × 10⁶, 2.15 × 10⁵, and 3.5 × 10⁴–2.15 × 10⁵ cfu mL^–1, respectively. The faecal coliform test revealed that only two out of twenty orange juice products had MPN of 2 and 20, the MPN of water ranged from 1–1800, while the effluent had MPN of 1800. The bacterial isolates that were isolated include E. coli, S. aureus, P. vulgaris, S. marcescens, S. pyogenes, B. cereus, B. subtilis, Micrococcus sp., Klebsiella sp., P. aeruginosa, and Enterobacter sp. Also, clinical and soil isolates of P. aeruginosa were used in the study. Among the eight antibiotics tested for resistance on five strains of each bacterium, seven different resistance patterns were observed among the bacterial isolates obtained from water, effluent and orange juice products. Among the clinical and soil isolates of P. aeruginosa, four multiple-drug resistance patterns were obtained. Thirty strains of E. coli and S. aureus were tested for -lactamase production and fourteen strains, seven each of E. coli and S. aureus that had high Minimum Inhibitory Concentration values (MIC) for both Amoxycillin and Cloxacillin were positive.     

Molecular characterization of antibiotic resistance in enterococci recovered from seagulls (Larus cachinnans) representing an environmental health problem

Antimicrobial resistance and the mechanisms implicated were studied in 54 enterococci recovered from 57 seagull fecal samples. Almost 78% of the recovered enterococci showed resistance against one or more antibiotics and these isolates were identified to the species level. E. faecium was the most prevalent species (52.4%). High percentages of erythromycin and tetracycline resistances were found among our isolates (95.2%), and lower percentages were identified to other antibiotics. Most of the tetracycline-resistant strains carried the tet(M) and/or tet(L) genes. Genes associated with Tn916/Tn1545 and/or Tn5397 transposons were detected in 45% of tetracycline-resistant isolates. The erm(B) gene was detected in 65% of erythromycin-resistant isolates. The vat(D) and vat(E) genes were present in 5.9% and 11.8% of quinupristin/dalfopristin-resistant isolates, respectively. The ant(6)-Ia gene was identified in 57.1% of streptomycin-resistant isolates. All nine kanamycin-resistant isolates carried the aph(3)'-IIIa gene. The cat(A) gene was found in two chloramphenicol-resistant isolates. Seagulls should be considered a risk species for spreading in the environment antimicrobial resistant enterococci and can serve as a sentinel for antibiotic pressure from the surrounding farm and urban setting.     

Perchlorate contamination of groundwater from fireworks manufacturing area in South India

Perchlorate contamination was investigated in groundwater and surface water from Sivakasi and Madurai in the Tamil Nadu State of South India. Sensitive determination of perchlorate (LOQ?=?0.005 μg/L) was achieved by large-volume (500 μL) injection ion chromatography coupled with tandem mass spectrometry. Concentrations of perchlorate were <0.005–7,690 μg/L in groundwater (n?=?60), <0.005–30.2 μg/L in surface water (n?=?11), and 0.063–0.393 μg/L in tap water (n?=?3). Levels in groundwater were significantly higher in the fireworks factory area than in the other locations, indicating that the fireworks and safety match industries are principal sources of perchlorate pollution. This is the first study that reports the contamination status of perchlorate in this area and reveals firework manufacture to be the pollution source. Since perchlorate levels in 17 out of 57 groundwater samples from Sivakasi, and none from Madurai, exceeded the drinking water guideline level proposed by USEPA (15 μg/L), further investigation on human health is warranted.     

Characterization of lead-resistant river isolate Enterococcus faecalis and assessment of its multiple metal and antibiotic resistance

Contamination of surface waters has a direct impact on the public health of entire communities. Microorganisms inhabiting contaminated surface waters have developed mechanisms of coping with a variety of toxic metals and drugs. Investigations were carried out to isolate and identify lead-resistant bacteria from the river K?z?l?rmak along the city of K?r?kkale, Turkey. Of the 33 lead-resistant isolates, one isolate with a minimal inhibitory concentration of 1,200 mg?L^?1 was isolated and identified as Enterococcus faecalis by using biochemical tests and 16S rRNA sequencing. Lead-resistant E. faecalis isolate was found out to be resistant to other heavy metals like aluminum, lithium, barium, chromium, iron, silver, tin, nickel, zinc, and strontium and to drugs like amikacin, aztreonam, and gentamicin. E. faecalis harbored four plasmids with the molecular sizes of 1.58, 3.06, 22.76, and 28.95 kb. Plasmid profile analyses of cured derivatives revealed that the lead resistance ability of E. faecalis was still existing despite the elimination of all the plasmids. Moreover, the antibiotic resistance pattern of the cured derivatives did not demonstrate any change from the parental strain. Our findings indicated that the lead resistance genes of E. faecalis were located on the chromosomal DNA rather than the plasmid.     

Relative proportions of <Emphasis Type="Italic">E. coli</Emphasis> and <Emphasis Type="Italic">Enterococcus spp.</Emphasis> may be a good indicator of potential health risks associated with the use of roof harvested rainwater stored in tanks

A total of 285 water samples were collected from 71 roof harvested rainwater tanks from four villages in different provinces over a two-year (2013–2014) period during the early (October to December) and late (January to March) rainy season. Water quality was evaluated based on Escherichia coli, faecal coliforms and Enterococcus spp. prevalence using the IDEXX Quanti-Tray quantification system. Real-Time PCR was used to analyse a subset of 168 samples for the presence of Shigella spp., Salmonella spp. and E. coli virulence genes (stx1, stx2 and eaeA). Escherichia coli were detected in 44.1% of the samples, Enterococcus spp. in 57.9% and faecal coliforms in 95.7%. The most prevalent E. coli concentrations in harvested rainwater were observed in 29.1% of samples and 22.5% for Enterococcus spp. and, were within 1–10 cfu/100 ml and 10–100 cfu/100 ml, respectively, whereas those for faecal coliforms (36.6%) were within 100–1000 cfu/100 ml. On average 16.8% of the samples had neither E. coli nor Enterococcus spp. detected, while 33.9% had only Enterococcus spp. and 23.7% had only E. coli. E. coli and Enterococcus spp. were detected together in 25.5% of the samples. Evaluation of samples for potential pathogenic bacteria showed all tested samples to be negative for the Shigella spp. ipaH gene, while five tested positive for Salmonella ipaB gene. None of the samples tested positive for the stx1 and stx2 genes, and only two tested positive for the eaeA gene. These findings are potentially useful in the development of a simplified risk assessment strategy based on the concentrations of indicator bacteria.     

Effects of field storage method on <Emphasis Type="Italic">E. coli</Emphasis> concentrations measured in storm water runoff

Storm water runoff is increasingly assessed for fecal indicator organisms (e.g., Escherichia coli, E. coli) and its impact on contact recreation. Concurrently, use of autosamplers along with logistic, economic, technical, and personnel barriers is challenging conventional protocols for sample holding times and storage conditions in the field. A common holding time limit for E. coli is 8 h with a 10 °C storage temperature, but several research studies support longer hold time thresholds. The use of autosamplers to collect E. coli water samples has received little field research attention; thus, this study was implemented to compare refrigerated and unrefrigerated autosamplers and evaluate potential E. coli concentration differences due to field storage temperature (storms with holding times ≤24 h) and due to field storage time and temperature (storms >24 h). Data from 85 runoff events on four diverse watersheds showed that field storage times and temperatures had minor effects on mean and median E. coli concentrations. Graphs and error values did, however, indicate a weak tendency for higher concentrations in the refrigerated samplers, but it is unknown to what extent differing die-off and/or regrowth rates, heterogeneity in concentrations within samples, and laboratory analysis uncertainty contributed to the results. The minimal differences in measured E. coli concentrations cast doubt on the need for utilizing the rigid conventional protocols for field holding time and storage temperature. This is not to say that proper quality assurance and quality control is not important but to emphasize the need to consider the balance between data quality and practical constraints related to logistics, funding, travel time, and autosampler use in storm water studies.