Ashok J. Tamhankar

Antibiotics and antibiotic-resistant bacteria in waters associated with a hospital in Ujjain, India

BackgroundConcerns have been raised about the public health implications of the presence of antibiotic residues in the aquatic environment and their effect on the development of bacterial resistance. While there is information on antibiotic residue levels in hospital effluent from some other countries, information on antibiotic residue levels in effluent from Indian hospitals is not available. Also, concurrent studies on antibiotic prescription quantity in a hospital and antibiotic residue levels and resistant bacteria in the effluent of the same hospital are few. Therefore, we quantified antibiotic residues in waters associated with a hospital in India and assessed their association, if any, with quantities of antibiotic prescribed in the hospital and the susceptibility of Escherichia coli found in the hospital effluent.MethodsThis cross-sectional study was conducted in a teaching hospital outside the city of Ujjain in India. Seven antibiotics - amoxicillin, ceftriaxone, amikacin, ofloxacin, ciprofloxacin, norfloxacin and levofloxacin - were selected. Prescribed quantities were obtained from hospital records. The samples of the hospital associated water were analysed for the above mentioned antibiotics using well developed and validated liquid chromatography/tandem mass spectrometry technique after selectively isolating the analytes from the matrix using solid phase extraction. Escherichia coli isolates from these waters were tested for antibiotic susceptibility, by standard Kirby Bauer disc diffusion method using Clinical and Laboratory Standard Institute breakpoints.ResultsCiprofloxacin was the highest prescribed antibiotic in the hospital and its residue levels in the hospital wastewater were also the highest. In samples of the municipal water supply and the groundwater, no antibiotics were detected. There was a positive correlation between the quantity of antibiotics prescribed in the hospital and antibiotic residue levels in the hospital wastewater. Wastewater samples collected in the afternoon contained both a higher number and higher levels of antibiotics compared to samples collected in the morning hours. No amikacin was found in the wastewater, but E.coli isolates from all wastewater samples were resistant to amikacin. Although ciprofloxacin was the most prevalent antibiotic detected in the wastewater, E.coli was not resistant to it.ConclusionsAntibiotics are entering the aquatic environment of countries like India through hospital effluent. In-depth studies are needed to establish the correlation, if any, between the quantities of antibiotics prescribed in hospitals and the levels of antibiotic residues found in hospital effluent. Further, the effect of this on the development of bacterial resistance in the environment and its subsequent public health impact need thorough assessment.

Geographical Variation in Antibiotic-Resistant Escherichia coli Isolates from Stool, Cow-Dung and Drinking Water

Little information is available on relationships between the biophysical environment and antibiotic resistance. This study was conducted to investigate the antibiotic resistance pattern of Escherichia coli isolated from child stool samples, cow-dung and drinking water from the non-coastal (230 households) and coastal (187 households) regions of Odisha, India. Susceptibility testing of E. coli isolates (n = 696) to the following antibiotics: tetracycline, ampicillin/sulbactam, cefuroxime, cefotaxime, cefixime, cotrimoxazole, amikacin, ciprofloxacin, norfloxacin and nalidixic acid was performed by the disk diffusion method. Ciprofloxacin minimum inhibitory concentration (MIC) values were determined for ciprofloxacin-resistant isolates (n = 83). Resistance to at least one antibiotic was detected in 90% or more of the E. coli isolates. Ciprofloxacin MIC values ranged from 8 to 32 µg/mL. The odds ratio (OR) of resistance in E. coli isolates from children’s stool (OR = 3.1, 95% CI 1.18–8.01), cow-dung (OR = 3.6, 95% CI 1.59–8.03, P = 0.002) and drinking water (OR = 3.8, 95% CI 1.00–14.44, P = 0.049) were higher in non-coastal compared to coastal region. Similarly, the co-resistance in cow-dung (OR = 2.5, 95% CI 1.39–4.37, P = 0.002) and drinking water (OR = 3.2, 95% CI 1.36–7.41, P = 0.008) as well as the multi-resistance in cow-dung (OR = 2.2, 95% CI 1.12–4.34, P = 0.022) and drinking water (OR = 2.7, 95% CI 1.06–7.07, P = 0.036) were also higher in the non-coastal compared to the coastal region.

Seasonal and Temporal Variation in Release of Antibiotics in Hospital Wastewater: Estimation Using Continuous and Grab Sampling

The presence of antibiotics in the environment and their subsequent impact on resistance development has raised concerns globally. Hospitals are a major source of antibiotics released into the environment. To reduce these residues, research to improve knowledge of the dynamics of antibiotic release from hospitals is essential. Therefore, we undertook a study to estimate seasonal and temporal variation in antibiotic release from two hospitals in India over a period of two years. For this, 6 sampling sessions of 24 hours each were conducted in the three prominent seasons of India, at all wastewater outlets of the two hospitals, using continuous and grab sampling methods. An in-house wastewater sampler was designed for continuous sampling. Eight antibiotics from four major antibiotic groups were selected for the study. To understand the temporal pattern of antibiotic release, each of the 24-hour sessions were divided in three sub-sampling sessions of 8 hours each. Solid phase extraction followed by liquid chromatography/tandem mass spectrometry (LC-MS/MS) was used to determine the antibiotic residues. Six of the eight antibiotics studied were detected in the wastewater samples. Both continuous and grab sampling methods indicated that the highest quantities of fluoroquinolones were released in winter followed by the rainy season and the summer. No temporal pattern in antibiotic release was detected. In general, in a common timeframe, continuous sampling showed less concentration of antibiotics in wastewater as compared to grab sampling. It is suggested that continuous sampling should be the method of choice as grab sampling gives erroneous results, it being indicative of the quantities of antibiotics present in wastewater only at the time of sampling. Based on our studies, calculations indicate that from hospitals in India, an estimated 89, 1 and 25 ng/L/day of fluroquinolones, metronidazole and sulfamethoxazole respectively, might be getting released into the environment per 100 hospital beds.

Detection of carbapenem resistance genes and cephalosporin, and quinolone resistance genes along with oqxAB gene in Escherichia coli in hospital wastewater: a matter of concern

This study was performed to detect the presence of Escherichia coli resistant to cephalosporins, carbapenems and quinolones in hospital wastewater.

Antibiotics in Wastewater of a Rural and an Urban Hospital before and after Wastewater Treatment, and the Relationship with Antibiotic Use—A One Year Study from Vietnam

Hospital effluents represent an important source for the release of antibiotics and antibiotic resistant bacteria into the environment. This study aims to determine concentrations of various antibiotics in wastewater before and after wastewater treatment in a rural hospital (60 km from the center of Hanoi) and in an urban hospital (in the center of Hanoi) in Vietnam, and it aims to explore the relationship between antibiotic concentrations in wastewater before wastewater treatment and quantities of antibiotics used in the rural hospital, over a period of one year in 2013. Water samples were collected using continuous sampling for 24 h in the last week of every month. The data on quantities of antibiotics delivered to all inpatient wards were collected from the Pharmacy department in the rural hospital. Solid-phase extraction and high performance liquid chromatography-tandem mass spectrometry were used for chemical analysis. Significant concentrations of antibiotics were present in the wastewater both before and after wastewater treatment of both the rural and the urban hospital. Ciprofloxacin was detected at the highest concentrations in the rural hospital’s wastewater (before treatment: mean = 42.8 µg/L; after treatment: mean = 21.5 µg/L). Metronidazole was detected at the highest concentrations in the urban hospital’s wastewater (before treatment: mean = 36.5 µg/L; after treatment: mean = 14.8 µg/L). A significant correlation between antibiotic concentrations in wastewater before treatment and quantities of antibiotics used in the rural hospital was found for ciprofloxacin (r = 0.78; p = 0.01) and metronidazole (r = 0.99; p < 0.001).

Antibiotic Resistance in an Indian Rural Community: A ‘One-Health’ Observational Study on Commensal Coliform from Humans, Animals, and Water

Antibiotic-resistant bacteria are an escalating grim menace to global public health. Our aim is to phenotype and genotype antibiotic-resistant commensal Escherichia coli (E. coli) from humans, animals, and water from the same community with a ‘one-health’ approach. The samples were collected from a village belonging to demographic surveillance site of Ruxmaniben Deepchand (R.D.) Gardi Medical College Ujjain, Central India. Commensal coliforms from stool samples from children aged 1–3 years and their environment (animals, drinking water from childrens households, common source- and waste-water) were studied for antibiotic susceptibility and plasmid-encoded resistance genes. E. coli isolates from human (n = 127), animal (n = 21), waste- (n = 12), source- (n = 10), and household drinking water (n = 122) carried 70%, 29%, 41%, 30%, and 30% multi-drug resistance, respectively. Extended spectrum beta-lactamase (ESBL) producers were 57% in human and 23% in environmental isolates. Co-resistance was frequent in penicillin, cephalosporin, and quinolone. Antibiotic-resistance genes blaCTX-M-9 and qnrS were most frequent. Group D-type isolates with resistance genes were mainly from humans and wastewater. Colistin resistance, or the mcr-1 gene, was not detected. The frequency of resistance, co-resistance, and resistant genes are high and similar in coliforms from humans and their environment. This emphasizes the need to mitigate antibiotic resistance with a ‘one-health’ approach.

Disinfection of Multidrug Resistant Escherichia coli by Solar-Photocatalysis using Fe-doped ZnO Nanoparticles

Spread of antibiotic resistant bacteria through water, is a threat to global public health. Here, we report Fe-doped ZnO nanoparticles (Fe/ZnO NPs) based solar-photocatalytic disinfection (PCD) of multidrug resistant Escherichia coli (MDR E. coli). Fe/ZnO NPs were synthesized by chemical precipitation technique, and when used as photocatalyst for disinfection, proved to be more effective (time for complete disinfection = 90 min) than ZnO (150 min) and TiO2 (180 min). Lipid peroxidation and potassium (K+) ion leakage studies indicated compromisation of bacterial cell membrane and electron microscopy and live-dead staining confirmed the detrimental effects on membrane integrity. Investigations indicated that H2O2 was the key species involved in solar-PCD of MDR E. coli by Fe/ZnO NPs. X-ray diffraction and atomic absorption spectroscopy studies showed that the Fe/ZnO NPs system remained stable during the photocatalytic process. The Fe/ZnO NPs based solar-PCD process proved successful in the disinfection of MDR E. coli in real water samples collected from river, pond and municipal tap. The Fe/ZnO NPs catalyst made from low cost materials and with high efficacy under solar light may have potential for real world applications, to help reduce the spread of resistant bacteria.

Climatic factors and community - associated methicillin-resistant Staphylococcus aureus skin and soft-tissue infections - a time-series analysis study.

Skin and soft tissue infections caused by Staphylococcus aureus (SA-SSTIs) including methicillin-resistant Staphylococcus aureus (MRSA) have experienced a significant surge all over the world. Changing climatic factors are affecting the global burden of dermatological infections and there is a lack of information on the association between climatic factors and MRSA infections. Therefore, association of temperature and relative humidity (RH) with occurrence of SA-SSTIs (n = 387) and also MRSA (n = 251) was monitored for 18 months in the outpatient clinic at a tertiary care hospital located in Bhubaneswar, Odisha, India. The Kirby-Bauer disk diffusion method was used for antibiotic susceptibility testing. Time-series analysis was used to investigate the potential association of climatic factors (weekly averages of maximum temperature, minimum temperature and RH) with weekly incidence of SA-SSTIs and MRSA infections. The analysis showed that a combination of weekly average maximum temperature above 33 °C coinciding with weekly average RH ranging between 55% and 78%, is most favorable for the occurrence of SA-SSTIs and MRSA and within these parameters, each unit increase in occurrence of MRSA was associated with increase in weekly average maximum temperature of 1.7 °C (p = 0.044) and weekly average RH increase of 10% (p = 0.097).

Quality of Water and Antibiotic Resistance of Escherichia coli from Water Sources of Hilly Tribal Villages with and without Integrated Watershed Management—A One Year Prospective Study

In many hilly tribal areas of the world, water scarcity is a major problem and diarrhoea is common. Poor quality of water also affects the environment. An integrated watershed management programme (IWMP) aims to increase availability of water and to improve life conditions. Globally, there is a lack of information on water contamination, occurrence of diarrhoea and antibiotic resistance, a serious global concern, in relation to IWMP in hilly tribal areas. Therefore, a prospective observational study was conducted during 2011–2012 in six villages in a hilly tribal belt of India, three with and three without implementation of an IWMP, to explore quality of water, diarrhoeal cases in the community and antibiotic resistance of Escherichia coli from water sources. The results showed that physico-chemical quality of water was within limits of safe consumption in all samples. The odds of coliform contamination in water samples was 2.3 times higher in non-watershed management villages (NWMV) compared to integrated watershed management villages (IWMV) (95% CI 0.8–6.45, p = 0.081). The number of diarrhoeal cases (18/663 vs. 42/639, p < 0.05) was lower in IWMV as compared to NWMV. Overall E. coli isolates showed high susceptibility to antibiotics. Resistance to a wider range of antibiotics was observed in NWMV.

Understanding Healthcare Workers Self-Reported Practices, Knowledge and Attitude about Hand Hygiene in a Medical Setting in Rural India.

Aim To describe self-reported practices and assess knowledge and attitudes regarding hand hygiene among healthcare workers in a rural Indian teaching hospital. Setting A rural teaching hospital and its associated medical and nursing colleges in the district of Ujjain, India. Method The study population consisted of physicians, nurses, teaching staff, clinical instructors and nursing students. Self-administered questionnaires based on the World Health Organization Guidelines on Hand Hygiene in Healthcare were used. Results Out of 489 healthcare workers, 259 participated in the study (response rate = 53%). The proportion of healthcare workers that reported to ‘always’ practice hand hygiene in the selected situations varied from 40–96% amongst categories. Reported barriers to maintaining good hand hygiene were mainly related to high workload, scarcity of resources, lack of scientific information and the perception that priority is not given to hand hygiene, either on an individual or institutional level. Previous training on the topic had a statistically significant association with self-reported practice (p = 0.001). Ninety three per cent of the respondents were willing to attend training on hand hygiene in the near future. Conclusion Self-reported knowledge and adherence varied between situations, but hand hygiene practices have the potential to improve if the identified constraints could be reduced. Future training should focus on enhancing healthcare workers’ knowledge and understanding regarding the importance of persistent practice in all situations.