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Occurrence of Antimicrobial Resistant Escherichia Coli in Waterways of Southeast Queensland, Australia

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Occurrence of Antimicrobial Resistant Escherichia Coli in Waterways of Southeast Queensland, Australia Medical Research Archives, Vol. 5, Issue 9, September 2017 Occurrence of antimicrobial resistant Escherichia coli in waterways of southeast Queensland, Australia Occurrence of antimicrobial resistant Escherichia coli in waterways of southeast Queensland, Australia Author: Abstract A.J. Watkinson1* Antimicrobial resistance is a global health issue. G.B. Micalizzi2 The discharge, maintenance and transfer of antimicrobial resistance to the aquatic 2 J.B. Bates environment and the risk this presents is relatively S.D. Costanzo3 unknown. This work describes the presence and distribution of antimicrobial resistant Escherichia Affiliations: coli in surface waters of seven rivers in south east Queensland, Australia. Resistance to four 1 School of Engineering antimicrobials (ampicillin, tetracycline, University of Queensland sulfamethoxazole and ciprofloxacin) was St. Lucia, Queensland determined using a method combining Australia 4072 chromogenic microbial detection with breakpoint antimicrobial resistance analysis. E. coli 2 Public Health Microbiology concentration and antimicrobial resistance was Queensland Health Forensic and significantly higher (p < 0.05) at sites receiving Scientific Services WWTP discharge compared to sites with no direct 39 Kessels Road WWTP discharge. There was a positive Coopers Plains Qld 4108 correlation (ρ < 0.001, r = 0.95) between E. coli 3University of Maryland Center for concentration and volume of WWTP discharge Environmental Science into these rivers. This would suggest that WWTPs 2020 Horns Point Road are the primary source of antimicrobial resistant Cambridge, MD, USA 21601 E. coli in the study region. There was no correlation (ρ > 0.05) between WWTP discharge *Address for Correspondence: volume into rivers and incidence of antimicrobial Dr Andrew Watkinson resistance for ampicillin, sulfamethoxazole and tetracycline. However, there was a positive Queensland Bulk Water Supply correlation (ρ < 0.01, r = 0.82) between incidence Authority (trading as Seqwater) of ciprofloxacin resistance and WWTP discharge 117 Brisbane Street, Ipswich, QLD, volume into rivers. This would suggest that E. coli 4305 (street Address) resistance to ciprofloxacin found in the study area PO Box 16146, City East, QLD 4002 is driven primarily by WWTP discharge, with (postal Address) resistance to other investigated antimicrobials Phone: +61 7 3035 5635 influenced by additional sources and/or drivers. Email: [email protected] Keywords: antimicrobial, antibiotic, resistance, wastewater, aquatic, environment, Australia Copyright 2017 KEI Journals. All Rights Reserved Page │1 Medical Research Archives, Vol. 5, Issue 9, September 2017 Occurrence of antimicrobial resistant Escherichia coli in waterways of southeast Queensland, Australia 1. Introduction Traditionally, the faecal coliform The World Health Organisation has Escherichia coli has been the foremost described antimicrobial resistance as one of indicator of faecal contamination in water the key global health issues and a profound quality monitoring. Escherichia coli is a threat to human health (1). Through overuse causative agent of a number of infections in and poor management of antimicrobials, we both humans (41, 42) and animals (43, 44), now find ourselves in the precarious mostly caused by a number of pathogenic strains, such as E. coli O157:H7 (45). situation where careful management of antimicrobials is essential to combat the Additionally, E. coli has been shown as a widespread emergence of resistant bacteria. significant reservoir of genes coding for Global responses to this have seen the antimicrobial drug resistance and therefore development of taskforces such as the WHO represents a useful indicator for and Australian Strategic and Technical antimicrobial resistance in bacterial Advisory Groups on Antimicrobial communities (46). Therefore studies of Resistance and the production of Strategic antimicrobial resistance patterns in E. coli Action Plans in an attempt to address the have implications on many levels since they issue (1, 2). can be used as an indicator for overall antimicrobial resistance and findings have While much of this attention has been direct impacts on risks to public health. directed towards management of antimicrobial use and monitoring the The aim of this study was to prevalence of bacterial resistance within the investigate the levels of antimicrobial community, antimicrobials and ARB in the resistance in E. coli isolated from surface aquatic environment have received waters in South-East Queensland. This information will contribute to our comparatively little attention. This is surprising given the reliance on our water understanding of antimicrobial resistance in resources and the potential for the spread the aquatic environment and the potential and maintenance of bacterial resistance to environmental and public health risk antimicrobials in this environment. associated with exposure to aquatic bacteria. Antimicrobials have recently been identified quite ubiquitously in the aquatic 2. Materials and Methods environment at sub-inhibitory concentrations 2.1. Preparation of media (3-7). However, knowledge of sub-inhibitory effects of antimicrobials on environmental Media was prepared as described in bacteria is scarce and contradictory (8-11). Watkinson et. al. 2007 (47). Four antimicrobials were chosen for susceptibility Antimicrobial resistant bacteria can be testing: ampicillin; tetracycline; considered a contaminant of emerging sulfamethoxazole; and ciprofloxacin due to concern through their regular discharge to their clinical relevance and their frequent the aquatic environment from wastewater identification in the aquatic environment (3, treatment plants (WWTPs) (12-18), 6, 48). aquaculture (19-24) and runoff from agricultural regions (25-31). Studies 2.2. Study Area investigating the prevalence of antimicrobial resistance in the aquatic environment are Water samples were collected on three limited, though they are starting to appear occasions from 57 sites (n = 171 samples) with increasing frequency (32-40). across nine rivers in South-East Queensland, Australia (Figure 1). These rivers comprise Copyright 2017 KEI Journals. All Rights Reserved Page │2 Medical Research Archives, Vol. 5, Issue 9, September 2017 Occurrence of antimicrobial resistant Escherichia coli in waterways of southeast Queensland, Australia of freshwater, estuarine and marine sections to discharges with PS sites directly receiving and discharge either into a large semi- discharge from a WWTP and NPS sites enclosed bay (Moreton Bay) or directly into having no direct input. Each site was the Pacific Ocean. Study sites were located sampled on three occasions over a 6-month within the estuarine-marine zones of these period. To assist in interpreting results, rivers and were classified as point source major practices within each river are (PS, 18 sites) or non-point source (NPS, 33 summarised in Table 1. sites) sites based on their proximity (< 50 m) Noosa River Maroochy River Mooloolah River Caboolture River Moreton Pacific Bay Ocean Pine River Lower Brisbane River N Logan/Albert River Point Source (PS) Sites Non-point Source (NPS) Sites 0 Kilometres 50 Nerang River Figure 1: Location of eight investigated river systems in South- East Queensland. Point source and non-point source sites are indicated. Copyright 2017 KEI Journals. All Rights Reserved Page │3 Medical Research Archives, Vol. 5, Issue 9, September 2017 Occurrence of antimicrobial resistant Escherichia coli in waterways of southeast Queensland, Australia Table 1: Characteristics of investigated rivers (82) a Catchment Dry Weather Total Length of Effluent Discharge River Area (km2) Population Streams (kms) Major Land Uses (ML d-1) Discharge Point Noosa 831 36,000 600 U, NB, CA, G 0 Pacific Ocean Maroochy 636 136,000 438 U, NB, G, IA 37 Pacific Ocean Mooloolah 223 83,000 322 U, NB, G, RR, MF 0 Pacific Ocean Caboolture 468 115,000 795 U, RR, G, IA 18 Moreton Bay Pine 816 177,000 553 U, RR, G, NB 35 Moreton Bay Lower Brisbane 1,171 888,000 845 U, RR, NB, G 282 Moreton Bay Logan/ Albert 4,133 407,000 2,828 RR, U, G, IA 50 Moreton Bay Nerang 498 455,000 928 U, RR, IA, NB 0b Pacific Ocean NB=Native Bush; CA=Conservation Area; G=Grazing; U=Urban; IA=Intensive Agriculture; RR= Rural Residential; MF=Managed Forests a SEQRWQMS (2001) b effluent discharge at river mouth on ebb tide 2.3. Susceptibility Testing control and each of the antimicrobial treatments for each site and plates were One litre of surface water was o collected at each site at approximately 0.5 m incubated at 35 C for 24 hrs. After depth in autoclaved glass amber jar and incubation, blue colonies were counted transported back to the laboratory on ice. A under ambient light and confirmed under dilution series of 100, 10, 1, 0.1 and 0.01 ml long-wave UV light (366 nm). For each antimicrobial, percent resistance was was filtered through 0.22 µm membrane filters (Millepore, Bedford, MA, USA), the calculated by directly comparing counts on latter three volumes combined with 10 ml the antimicrobial plate with corresponding peptone. All filters were rinsed with peptone. counts on the control plate: A dilution series was prepared for the [E.coli] antibiotic plate % Resistance = x 100 (1) [E.coli] control plate Escherichia coli ATCC 25922 was with total riverine effluent discharge and chosen as a control strain carrying no river catchment population investigated for resistance to selected antimicrobials. correlation with median riverine
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