Commensal bacteria are the representative of the reservoir of antibiotic resistance genes present in a community. The usage of antibiotics along with the demographic factors is generally associated with an increase in antibiotics resistance in pathogens. Northeast (NE) India is untapped with regard to antibiotic resistance prevalence and spread. In the current study, the prevalence of antibiotic-resistant commensal Escherichia coli in pre-school and school-going children (n = 550, 1–14 years old) from the rural areas of the state of Sikkim—an NE Indian state, with respect to associated demographic factors was investigated. A total of 550 fecal E. coli isolates were collected during July 2015 to June 2017. A structured questionnaire was used to collect data to ascertain the potential factors associated with the carriage of antibiotic resistance E. coli among the children. Statistical analysis along with a logistic regression identified potential external factors affecting the observed antibiotic resistance pattern. The data indicated a high prevalence of resistance to common antibiotics like ampicillin (92%), ceftazidime (90%), cefoxitin (88%), streptomycin (40%) and tetracycline (36%), but no resistance to chloramphenicol. The resistance to the combination of penicillin and quinolone group of antibiotics was observed in fifty-two percent of the isolates. A positive correlation between the harboring of antibiotics resistant E. coli with different demographic factors was observed such as, with children living in nuclear family (vs joint family 63.15%, OR 0.18, 95% CI:0.11–0.28, p < 0.01), below higher secondary maternal education (vs college graduates 59.27% OR 0.75, 95% CI:0.55–1.02, p < 0.02). A close association between different demographic factors and the high prevalence of antibiotic-resistant commensal E. coli in the current study suggests a concern over rising misuse of antibiotics that warrants a future threat of emergence of multidrug-resistant pathogen isolates.

Continuous decline in potable water sources has raised serious concerns over human health. Developing countries are the most affected in this regard due to a lack of proper hygiene maintenance. Sikkim, an Eastern Himalayan state with mountains as the predominant topological features, harbors several perennial natural springs. Spring water is the primary source of potable water for the population in four districts of the state viz. East, West, North and South. Recent outbreaks of water-borne diseases and the relative lack of scientific studies on its potential correlation with the water quality of the area have educed this study. Physicochemical parameters of springs, community reservoirs, and household water were analyzed by ICP-MS and multi probe meter. Using the membrane filtration method, the microbial quality of the water samples during different seasons was assessed, primarily evaluating the presence of fecal indicators viz. Escherichia coli , total coliform and Enterococcus . The seasonal risk category of the water sources was also determined. Most of the physicochemical parameters of the spring water were within the permissible limits of WHO standards. However, water from four districts was recorded with traces of toxic heavy metals like mercury (0.001–0.007 mg/l), lead (0.001–0.007 mg/l), and selenium (0.526–0.644 mg/l), which are above the permissible limits of WHO. All the spring water samples were categorized as Mg-HC type and can be predicted as shallow fresh ground water based on the piper analysis. Microbial confirmatory testing indicated severe fecal contamination of water sources with high counts of total coliform (TC), Escherichia coli (EC) and Enterococcus (EN). The highest level of TC was recorded from West Sikkim (37.26 cfu/100 ml) and the lowest in North Sikkim (22.13 cfu/100 ml). The highest level of contamination of E. coli and Enterococcus was found in East Sikkim (EC = 8.7 cfu/100 ml; EN = 2.08 cfu/100 ml) followed by South Sikkim (EC = 8.4 cfu/100 ml; EN = 2.05 cfu/100 ml). There was a significant positive correlation between the contamination levels of the spring water and the community reservoir tank. As far as the seasonal variation is concerned, the rainy season showed the most contamination with coliform correlating with a high incidence of different water-borne diseases (East = 86%; West = 100%; South = 100%; North = 80%).

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Every year millions of people die due to fatal waterborne diseases around the world especially in developing countries like India. Sikkim, a northeastern state of India, greatly depends on natural water sources. About 80% of the population of Sikkim depends on natural spring water for domestic as well as agricultural use. Recent waterborne disease outbreaks in the state raises a concerning question on water quality. In this study, we analyzed water quality especially for the detection of Enterobacteriaceae members from four districts of the state. Isolation with selective culture media techniques and taxonomic characterization of Enterobacteriaceae bacteria with 16S rRNA gene showed the prevalence of Escherichia coli (37.50%), Escherichia fergusonii (29.41%), Klebsiella oxytoca (36.93%), Citrobacter freundii (37.92%), Citrobacter amalonaticus (43.82%), Enterobacter sp. (43.82%), Morganella morganii (43.82%), Hafnia alvei (32.42%), Hafnia paralvei (38.74%), and Shigella flexneri (30.47%) in the spring water of Sikkim. Antibiotic susceptibility test (AST) showed resistance of the isolates to common antibiotics like ampicillin, amoxicillin as well as to third generation antibiotics like ceftazidime and carbapenem. None of the isolates showed resistance to chloramphenicol. E. coli isolated from spring water of Sikkim showed presence of different virulence genes such as stx1 (81.81%), elt (86.66%), and eae (66.66%) along with resistance gene for ampicillin (CITM) (80%), quinolones (qnrB) (44.44%), tetracycline (tetO) (66.66%), and streptomycin (aadA1) (66.66%). The data indicates a high incidence rate of multiple antibiotic resistant enteric bacteria in the spring water of Sikkim. Additionally, the presence of enteric bacteria in the water samples indicates widespread fecal contamination of the spring water.

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