A Three-Year Follow-Up Study of Antibiotic and Metal Residues, Antibiotic Resistance and Resistance Genes, Focusing on Kshipra—A River Associated with Holy Religious Mass-Bathing in India: Protocol Paper

Abstract

Background: Antibiotic resistance (ABR) is one of the major health emergencies for global society. Little is known about the ABR of environmental bacteria and therefore it is important to understand ABR reservoirs in the environment and their potential impact on health. Method/Design: Quantitative and qualitative data will be collected during a 3-year follow-up study of a river associated with religious mass-bathing in Central India. Surface-water and sediment samples will be collected from seven locations at regular intervals for 3 years during religious mass-bathing and in absence of it to monitor water-quality, antibiotic residues, resistant bacteria, antibiotic resistance genes and metals. Approval has been obtained from the Ethics Committee of R.D. Gardi Medical College, Ujjain, India (No. 2013/07/17-311). Results: The results will address the issue of antibiotic residues and antibiotic resistance with a focus on a river environment in India within a typical socio-behavioural context of religious mass-bathing. It will enhance our understanding about the relationship between antibiotic residue levels, water-quality, heavy metals and antibiotic resistance patterns in Escherichia coli isolated from river-water and sediment, and seasonal differences that are associated with religious mass-bathing. We will also document, identify and clarify the genetic differences/similarities relating to phenotypic antibiotic resistance in bacteria in rivers during religious mass-bathing or during periods when there is no mass-bathing.

Abundance and diversity of phages, microbial taxa and antibiotic resistance genes in the sediments of the river Ganges through metagenomic approach

“…Furthermore, study from Diwan et al reported the presence of ARG types (beta-lactams, quinolone etc.) from seven regular and three mass-bathing sampling points from the river Kshipra, Ujjain, India [91]. These findings from different studies irrespective of distinct source rivers represent commonality and reported to have the presence of mainly beta-lactam and tetracycline resistance genes.…”

Mass bathing events in River Kshipra, Central India- influence on the water quality and the antibiotic susceptibility pattern of commensal E.coli

“…The present study is a part of an ongoing project, which has been described in detail previously [15]. In brief, the study samples were collected from River Kshipra, in its 93 km flow through Ujjain district, before, during and after mass-bathing events.…”

“…The pre-bathing samples were collected one day prior of the bathing event, post-bathing samples were collected 6-7 hours after bathing event while during-bathing samples were collected at the peak hours of the bathing event, generally between 9 to 11AM. Each time, riverwater and -sediment were collected in sterile screw-capped bottles and conical falcon tube respectively as described in detail [15]. Approximately 2Kg of sediment and 3.5L river-water were collected for analysis of water quality parameters, colony counts and antibacterial susceptibility as described elsewhere [15].…”

“…Each time, riverwater and -sediment were collected in sterile screw-capped bottles and conical falcon tube respectively as described in detail [15]. Approximately 2Kg of sediment and 3.5L river-water were collected for analysis of water quality parameters, colony counts and antibacterial susceptibility as described elsewhere [15]. All the collected samples were stored in icebox at 4-6˚C and transported to central research laboratory of R D Gardi Medical College, Ujjain within one hour.…”

“…The physical and chemical parameters (ambient and water temperature, pH, conductivity, total dissolved solids (TDS) free carbon dioxide (free CO 2 ), carbonate alkalinity and dissolved oxygen (DO)) of collected river-water and river-sediment samples were analyzed in field [15]. After transport to the laboratory, samples were analyzed for turbidity, hardness, chloride, alkalinity, nitrate nitrogen available phosphorous, total suspended solids (TSS), biochemical oxygen demand (BOD), chemical oxygen demand (COD) and total phosphorous [15].…”