Faecal Coliform Bacteria in Febros River (Northwest Portugal): Temporal Variation, Correlation with Water Parameters, and Species Identification

Febros river water was sampled weekly, during 35 successive weeks, and analyzed for microbiological (total coliforms, faecal coliforms, faecal streptococci and enterococci) and chemical-physical (ammonia and temperature) parameters. All microbiological parameters were highly correlated with each other and with ammonia, suggesting that the simultaneous determination of all variables currently in use in the evaluation of the microbiological quality of waters is probably redundant, and could be simplified, and that ammonia should be tested as a sentinel parameter of the microbiological pollution load of Febros river. From the strains isolated from positive tubes of the faecal coliforms test (multiple tube fermentation technique) and retested in this assay, Escherichia coli, Klebsiella oxytoca and Klebsiella pneumoniae subsp. pneumoniae strains were positive, indicating that the faecal coliforms test is not totally specific for Escherichia coli, and can detect other bacteria. Considering that these Klebsiella spp. are not necessarily of faecal origin, it was concluded that the faecal coliforms test can overestimate true faecal pollution. From the strains isolated from positive tubes of the faecal coliforms procedure, only Escherichia coli strains were clearly positive in the β-D-glucuronidase test. All other species were negative or very weakly positive, suggesting that the assay of the β-D-glucuronidase activity is less prone to false positives than the faecal coliforms test in the quantification of Escherichia coli in environmental waters.     

Physical and Sanitary Quality of Hand-Dug Well Water from Oil-Producing Area of Nigeria

The physical (pH, Dissolved Solids, Dissolved Oxygen, conductivity, water depth temperature) and bacteriological (Total coliforms, Faecal coliforms) characteristics of 22 hand-dug wells in oil-rich Niger Delta of Nigeria were investigated. All coliforms counts (Log₁₀ MPN/100 ml) varied from well to well ranging from 0 to 6.0 although faecal coliform counts were markedly lower (0–3.5 vs 1.0–6.0). When both seasons were compared, faecal coliform counts were significantly higher (t test; P < 0.05) during the wet season whereas no significant difference was observed in respect of total coliforms. Unlike the total coliforms, faecal coliform counts increased in wells close to septic tanks during the wet season. Although a declining trend was observed during the wet season, the values of the physical parameters were within WHO limits. The pH was slightly acidic (Mean ± SD: dry season, 6.04 ± 0.66; wet, 6.43 ± 0.62) and may be attributed to emissions from gas flaring and petroleum refining activities, which is common in the area. While the total coliform counts reflected the unsanitary habits of the well users, the detection of faecal coliforms exceeding recommended numbers in 63% of the wells render them unfit for domestic use. The public health implication is obvious considering the increasing number of people using the wells due to the rapid urbanization of the oil-rich region.     

Efficiency of Modified H2S Test for Detection of Faecal Contamination in Water

Faecal contamination is a major causative factor for incidence of water borne infectious diseases. Certain hydrogen sulfide (H₂S) producing enteric bacteria such as Salmonella sp. and Citrobacter sp., assosiated with coliforms, have been considered for rapid detection of recent faecal contamination in water. The basic H₂S test medium, modified by adding 0.25 gm/L of L-cystine HCl, was examined for its efficiency with 90 water samples collected from 40 pipe supplies, 20 open wells, 15 hand pumps and 15 different surface water bodies (river, streams and ponds). Sterilized modified culture medium in glass vials was inoculated with 100 mL of each sample and incubated at 20, 25, 30, 35 and 44 ^∘C for 18, 24, 42, 48, 66 and 72 h. Blackening of content in incubated vials was considered positive. For comparison, most probable number (MPN) of coliform and faecal coliform per 100 mL was also estimated in each sample by multiple tube fermentation (MTF) method. H₂S positive result was exhibited by 78% of samples. Coliform (> 10) and faecal coliform/100 mL were also detected in 59% of samples. Maximum H₂S positive results (100%) were found with well and surface water samples incubated at 30, 35 and 44 ^∘C for 18 h. Coliform (> 10) and faecal coliform/100 mL were also detected in most of these samples. Pipe supplies (60%) and hand pumps (73%) also exhibited considerable H₂S production. Coliforms and faecal coliforms were also found in significant number of these samples. Thus, the modified H₂S test may prove a useful alternative indicator of faecal contamination for water quality surveillance and screening of large number of water samples in short duration, particularly during any outbreak of water borne disease among rural population.     

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.     

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.     

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.     

Prevalence of heavy metal resistance in bacteria isolated from tannery effluents and affected soil

In the present study, a total of 198 bacteria were isolated, 88 from the tannery effluents and 110 from agricultural soil irrigated with the tannery effluents. Tannery effluents and soils were analyzed for metal concentrations by atomic absorption spectrophotometer. The tannery effluents and soil samples were found to be contaminated with chromium, nickel, zinc, copper, and cadmium. All isolates were tested for their resistance against Cr^6?+?, Cr^3?+?, Ni^2?+?, Zn^2?+?, Cu^2?+?, Cd^2?+?, and Hg^2?+?. From the total of 198 isolates, maximum bacterial isolates were found to be resistant to Cr^6?+? 178 (89.9%) followed by Cr^3?+? 146 (73.7%), Cd^2?+? 86 (43.4%), Zn^2?+? 83 (41.9%), Ni^2?+? 61 (30.8%), and Cu^2?+? 51 (25.6%). However, most of the isolates were sensitive to Hg^2?+?. Among the isolates from tannery effluents, 97.8% were resistant to Cr^6?+? and 64.8% were resistant to Cr^3?+?. Most of the soil isolates were resistant against Cr^6?+? (83.6%) and Cr^3?+? (81.8%). All isolates were categorized into Gram-positive and Gram-negative bacteria. In a total of 114 Gram-positive isolates, 91.2% were resistant to Cr^6?+? followed by 73.7% to Cr^3?+?, 42.1% to Zn^2?+?, 40.4% to Cd^2?+?, and 32.5% to Ni^2?+?. Among Gram-negative isolates, 88.1% were found showing resistance to Cr^6?+?, 75.0% to Cr^3?+?, and 47.6% were resistant to Cd^2?+?. Majority of these metal-resistant isolates were surprisingly found sensitive to the ten commonly used antibiotics. Out of 198 isolates, 114 were found sensitive to all antibiotics whereas only two isolates were resistant to maximum eight antibiotics at a time. Forty-one and 40 isolates which constitute 20.7% and 20.2% were resistant to methicilin and amoxicillin, respectively.     

Bacteriological safety assessment,hygienic habits and cross-contamination risks in a Nigerian urban sample of household kitchen environment

Urban household kitchen environment was assessed for safety by determining their levels of indicator bacteria, hygienic habits and risk of cross-contamination. Household kitchens (60) were selected in Warri Town, Nigeria, by the multi-stage sampling technique. Contact surfaces, water and indoor kitchen air were analysed for aerobic plate counts, total and faecal coliforms using Nutrient and McConkey media by swab/rinse method, membrane filtration and sedimentation methods, respectively. Hygienic habits and risk of cross-contamination were assessed with structured questionnaire which included socio-demographic variables. On the basis of median counts, the prevalence of high counts (log cfu/cm²/m³/100 mL) of aerobic plate counts (>3.0), total coliforms (>1.0) and faecal coliforms (>0) on contact surfaces and air was high (58.0–92.0%), but low in water (30.0–40.0%). Pots, plates and cutleries were the contact surfaces with low counts. Prevalence of poor hygienic habits and high risk of cross-contamination was 38.6 and 67.5%, respectively. Education, occupation and kitchen type were associated with cross-contamination risk (P = 0.002–0.022), while only education was associated with hygienic habits (P = 0.03). Cross-contamination risk was related (P = 0.01–0.05) to aerobic plate counts (OR 2.30; CL 1.30–3.17), total coliforms (OR 5.63; CL 2.76–8.25) and faecal coliforms (OR 4.24; CL 2.87–6.24), while hygienic habit was not. It can be concluded that urban household kitchens in the Nigerian setting are vulnerable to pathogens likely to cause food-borne infections.     

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.     

Seasonal and spatio-temporal distribution of faecal-indicator bacteria in Tyume River in the Eastern Cape Province, South Africa

We assessed the incidence of faecal-indicator bacteria in Tyume River over a 12-month period between August 2010 and July 2011. Total coliforms, faecal coliforms and enterococci were determined by the membrane filtration method. Total coliforms were detected in counts ranging from 2.1?×?10² to 3.4?×?10⁴?CFU/100 ml. Faecal coliform counts ranged from 1?×?10² to 1.6?×?10⁴?CFU/100 ml while enterococci counts were in the range of 3.3?×?10¹ to 5.1?×?10³?CFU/100 ml. Indicator bacteria counts increased from upstream to downstream sampling sites. Counts of indicator bacteria at all sites were significantly affected by seasonal changes. The bacteriological qualities of the river water were poor, exceeding the guideline of 200 CFU/100 ml and 33 CFU/100 ml for faecal coliforms and enterococci, respectively, for recreational water. Faecal coliform counts also exceeded the 1,000 CFU/100 ml guideline for water used in fresh produce irrigation. Microbial source tracking results showed that faecal pollution was predominantly of human origin during spring at all sampling sites. During other seasons, human faecal pollution was largely confined to midstream and downstream sampling sites. Generally, the presence of faecal-indicator bacteria in the river water samples suggests faecal pollution of this freshwater resource, raising the possibility of the presence of pathogenic microorganisms in the water and a threat to public health.     

Quantitative Analyses of Pollution-Indicator and Pathogenic Bacteria in Mumbai Waters from Ballast Water Exchange Perspective

As large numbers of ships either take in ballast from or discharge their ballast into the Mumbai Harbor region, it is pertinent to quantify microorganisms of health concerns and suggest on the suitability of water for ballasting purposes. To meet with this main aim, a selected set of general and pathogenic bacterial groups and their seasonal variations were studied from the Mumbai Harbor area. Sampling was carried out during postmonsoon (November 2001), pre-monsoon (April 2002) and monsoon (October 2002) periods. We quantified total coliforms and eight other groups of known human pathogenic bacteria from water, sediment, marine plant and animal samples collected in and around Mumbai Harbor. When compared with similar studies from other parts/harbors of the world, the Mumbai Harbor area has over 100-times higher levels of coliforms. The serotypes of Escherichia coli O157 and Shigella-Alkaligens Dispar group were abundant throughout the year. Even Vibrio cholerae, V. parahaemolyticus, Salmonella spp., campylobacters and aeromonads were present in large numbers. This comprehensive study, conducted with a view to quantify these bacterial groups and to evaluate the suitability of these waters for ballasting purposes, clearly suggests that both ballasting and deballasting be avoided in this region and alternative procedures should be developed for treating/handling ballast water.     

Applicability of universal Bacteroidales genetic marker for microbial monitoring of drinking water sources in comparison to conventional indicators

Water quality monitoring is essential for the provision of safe drinking water. In this study, we compared a selection of fecal indicators with universal Bacteroidales genetic marker to identify fecal pollution of a variety of drinking water sources. A total of 60 samples were collected from water sources. The microbiological parameters included total coliforms, fecal coliforms, Escherichia coli and fecal streptococci as the fecal indicator bacteria (FIB), Clostridium perfringens and H₂S bacteria as alternative indicators, universal Bacteroidales genetic marker as a promising alternative fecal indicator, and Salmonella spp., Shigella spp., and E. coli O157 as pathogenic bacteria. From 60 samples analyzed, Bacteroidales was the most frequently detected indicator followed by total coliforms. However, the Bacteroidales assay failed to detect the marker in nine samples positive for FIB and other alternative indicators. The results of our study showed that the absence of Bacteroidales is not necessarily an evidence of fecal and pathogenic bacteria absence and may be unable to ensure the safety of the water. Further research, however, is required for a better understanding of the use of a Bacteroidales genetic marker as an indicator in water quality monitoring programs.     

Isolation and Identification of Enterococci from Seawater Samples: Assessment of Their Resistance to Antibiotics and Heavy Metals

A hundred Enterococcus strains were isolated from seawater samples collected from coastal areas of Istanbul. Isolates were identified to the species level using standard biochemical tests specified by Facklam and Collins. The species distribution was as follows Enterococcus faecalis (96%), Enterococcus gallinarum (3%) and Enterococcus solitarius (1%). The resistance of bacteria to both heavy metals (zinc [Zn], iron [Fe], cadmium [Cd], chrome [Cr], cobalt [Co]) and antibiotics (ampicillin 10 μg [AP], penicillin G 10 Units [PG], gentamycin 10 μg [GM], streptomycin 10 μg [S], chloramphenicol 10 μg [C], erythromycin 15 μg [E], kanamycin 30 μg [K], amikacin 30 μg [AK], nalidixic acid 30 μg [NA], and vancomycin 30 μg [VA]) was evaluated. None of the strains was resistant to VA. It was found that among the 100 isolates, those that exhibit resistance to antibiotics, particularly NA, S and K, were also resistant all the heavy metals tested. To our knowledge this is the first report focusing on determination of resistance of environmental enterococci found in Istanbul against heavy metals and antibiotics. Thus, combined expressions of antibiotic and heavy metal resistance may help to reinforce ecological and epidemiological studies and to determine the role of these strains in antibiotic and heavy metal resistance dissemination.     

Physicochemical and microbiological assessment of recreational and drinking waters

The present study was aimed to make an assessment of health risk due to pollution and human pathogenic bacteria associated with the recreational and drinking water sources in twin densely populated holy Indian cities Ayodhya and Faizabad. Though physicochemical studies revealed that the water available in the area is under recommended limits for human use, it is unsafe on account of poor microbiological quality of surface and ground water in the region. The most probable number (MPN) test results revealed the preponderance of ≥2,400 total coliforms (TC) (100 ml)^− 1 in river, pond, dug well and kund waters. Contrary to that, 94% tube wells, 32% hand pumps and 25% piped supply water were under safe limits having <3 TC (100 ml)^− 1. The shallow depth (~40 ft), water logging and presence of septic tanks in the near vicinity are the possible reasons of poor microbial quality of hand pump drinking water. The municipal supply water passes along sewage line where loose connections and/or cracks in pipe lead to mixing and contamination. The significant best quality of tube well water evident from the absence of TC could be attributed to the depth of well ≥150 ft and usually their location away from the habitation. A total of 263 bacteria from 186 water samples were isolated, and at least five genera of enteric bacteria from various water sources were identified morphologically and biochemically as Escherichia coli, Klebsiella sp., Enterobacter sp., Shigella sp. and Salmonella sp. The serotyping of 72 E. coli and 36 Salmonella sp. revealed 51 as E. coli O157 and 20 as Salmonella sp. The presence of enteric pathogens in water sources pose threat to human health and therefore call for immediate remedial measures.     

Identifying fecal pollution sources using 3M™ Petrifilm™ count plates and antibiotic resistance analysis in the Horse Creek Watershed in Aiken County,SC (USA)

Sources of fecal coliform pollution in a small South Carolina (USA) watershed were identified using inexpensive methods and commonly available equipment. Samples from the upper reaches of the watershed were analyzed with 3M^? Petrifilm^? count plates. We were able to narrow down the study’s focus to one particular tributary, Sand River, that was the major contributor of the coliform pollution (both fecal and total) to a downstream reservoir that is heavily used for recreation purposes. Concentrations of total coliforms ranged from 2,400 to 120,333 cfu/100 mL, with sharp increases in coliform counts observed in samples taken after rain events. Positive correlations between turbidity and fecal coliform counts suggested a relationship between fecal pollution and stormwater runoff. Antibiotic resistance analysis (ARA) compared antibiotic resistance profiles of fecal coliform isolates from the stream to those of a watershed-specific fecal source library (equine, waterfowl, canines, and untreated sewage). Known fecal source isolates and unknown isolates from the stream were exposed to six antibiotics at three concentrations each. Discriminant analysis grouped known isolates with an overall average rate of correct classification (ARCC) of 84.3 %. A total of 401 isolates from the first stream location were classified as equine (45.9 %), sewage (39.4 %), waterfowl (6.2 %), and feline (8.5 %). A similar pattern was observed at the second sampling location, with 42.6 % equine, 45.2 % sewage, 2.8 % waterfowl, 0.6 % canine, and 8.8 % feline. While there were slight weather-dependent differences, the vast majority of the coliform pollution in this stream appeared to be from two sources, equine and sewage. This information will contribute to better land use decisions and further justify implementation of low-impact development practices within this urban watershed.     

Identification of source of faecal pollution of Tirumanimuttar River, Tamilnadu, India using microbial source tracking

Efficient management of deteriorating water bodies can be achieved by determining the sources of faecal pollution. Resourceful techniques for discrimination of the sources of Escherichia coli in surface water have recently been developed, including the use of river water to facilitate faecal indicator surveillance, identification of sources of faecal contamination and employing relevant management practices to maintain water quality. This study was conducted to employ microbial source tracking (MST) techniques for the determination of the sources of faecal pollution based on a water quality investigation of the physico-chemical characteristics and coliform count point of the Tirumanimuttar River. To accomplish this, an MST library-based antibiotic resistance analysis, serotyping and the genomic tool rep-PCR techniques were applied, and the obtained results were analysed statistically. Among 135 and 70 E. coli isolates present in the library and water samples collected from the river and nearby well water sources, respectively, most showed intrinsic, high or moderate resistance to antibiotics. Isolates from human and pig faecal sources were 92% homologous with the samples from the river, whereas isolates from sewage and dairy cattle showed 89% and 80% homology, respectively. These findings indicated that the Tirumanimuttar River is subjected to stress from anthropogenic activities and runoff contaminated with agricultural and human faecal contamination. The sources of faecal pollution identified in this study may facilitate the monitoring and management of the Tirumanimuttar River.     

Determination of antimicrobial and heavy metal resistance profiles of some bacteria isolated from aquatic amphibian and reptile species

The aim of the present study was to determine the level of antibiotic resistance patterns and distribution of heavy metal resistance of bacterial isolates from aquatic animals (Lissotriton vulgaris, Pelophylax ridibundus, Emys orbicularis, Mauremys rivulata, and Natrix natrix) in Turkey (Kavak Delta). A total of 153 bacteria have been successfully isolated from cloaca and oral samples of the aquatic amphibians and reptilians which were found, namely, Aeromonas sp. (n?=?29), Plesiomonas sp. (n?=?7), Vibrio sp. (n?=?12), Citrobacter sp. (n?=?12), Enterobacter sp. (n?=?11), Escherichia sp. (n?=?22), Klebsiella sp. (n?=?22), Edwardsiella sp. (n?=?6), Hafnia sp. (n?=?1), Proteus sp. (n?=?19), Providencia sp. (n?=?8), and Pseudomonas sp. (n?=?4). In terms of antibiotic and heavy metal susceptibility testing, each isolate was tested against 12 antibiotics and 4 metals. There was a high incidence of resistance to cefoxitin (46.40 %), ampicillin (44.44 %), erythromycin (35.29 %), and a low incidence of resistance to gentamicin (6.53 %), kanamycin (8.49 %), chloramphenicol (9.15 %), and cefotaxime (10.45 %). The multiple antibiotic resistance index of each bacterial species indicated that bacteria from raised amphibians and reptiles have been exposed to tested antibiotics, with results ranging from 0 to 0.58. Most isolates showed tolerance to different concentrations of heavy metals, and minimal inhibition concentrations ranged from100 to >3,200 μg/mL. According to these results, a significant occurrence of bacteria in the internal organs of reptiles and amphibians, with a high incidence of resistance against antibiotics and heavy metals, may risk aquatic animals and the public health. These data appoint the importance of epidemiological surveillance and microbiological monitoring and reinforce the need to implement environment protection programs for amphibian and reptile species.     

Monitoring E. coli and total coliforms in natural spring water as related to recreational mountain areas

Natural spring water has unique properties, as it is rich in minerals that are considered to be beneficial to human health. A survey of the microbiological quality of natural spring water was conducted to assess possible risks from the consumption of the water by visitors in recreational mountain areas located in Seoul, South Korea. The densities of total coliforms and Escherichia coli were measured during the spring and the summer of 2002 to investigate the presence of coliform bacteria in the drinking spring waters. Total coliforms were detected in all samples and the mean density of total coliforms was up to a maximum of 228 CFU/mL. Detectable E. coli was found in 78% of all samples and the mean densities of E. coli varied from a minimum of 0 CFU/mL to a maximum of 15 CFU/mL in all samples. Malfunctioning septic systems and wildlife population appear to be the main source of E. coli contamination. Presence of E. coli in natural spring water indicates potential adverse health effects for individuals or populations exposed to this water. The fecal contaminated spring water may present an unacceptable risk to humans if it is used as raw drinking water.     

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.     

Impact of long-term wastewater application on microbiological properties of vadose zone

Impact of wastewater irrigation on some biological properties was studied in an area where treated sewage water is being supplied to the farmers since 1979 in the western part of National Capital Territory of New Delhi under Keshopur Effluent Irrigation Scheme. Three fields were selected which had been receiving irrigation through wastewater for last 20, 10 and 5 years. Two additional fields were selected in which the source of irrigation water was tubewell. The soil bacterial and fungal population density was studied in soil layers of 0?C15, 15?C30, 30?C60 and 60?C120 cm depths. Groundwater samples were collected from the piezometers installed in the field irrigated with sewage water for last 20, 10 and 5 years. Results indicate that there was significant increase in bacterial and fungal count in sewage-irrigated soils as compared to their respective control. The population density of bacteria and fungi in waste water-irrigated soils increased with the duration of sewage water application and decreased with increasing depth. The bacterial and fungal count was also directly proportional to organic carbon, sand and silt content and negatively correlated to the clay content, electrical conductivity, pH and bulk density of the soil. Groundwater under sewage-irrigated fields had higher values of most probable number (MPN) index as compared to that of tubewell water-irrigated fields. All the shallow and deep groundwaters were found to be contaminated with faecal coliforms. The vadose zone had filtered the faecal coliform to the tune of 98?C99%, as the MPN index was reduced from ??18,000 per 100 ml of applied waste water to 310 per 100 ml of groundwater under 20 years sewage-irrigated field. The corresponding values of MPN were 250 and 130 per 100 ml of shallow groundwater under 10 and 05 years sewage-irrigated fields, respectively. Rapid detection of faecal contamination suggested that the Citrobacter freundii and Salmonella were dominant in shallow groundwater, while Escherichia coli was dominant in deep groundwater collected from sewage-irrigated field.