Medical Research Archives, Vol. 5, Issue 9, September 2017 Occurrence of antimicrobial resistant Escherichia coli in waterways of southeast ,

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 , 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 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

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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 () 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

Caboolture River Moreton Pacific Bay Ocean

Lower

N Logan/Albert River Point Source (PS) Sites Non-point Source (NPS) Sites

0 Kilometres 50

Figure 1: Location of eight investigated river systems in South- East Queensland. Point source and non-point source sites are indicated.

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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 E. coli Ambient E. coli concentrations were concentration, and antimicrobial resistance. calculated from control plate results for each site. A one-way ANOVA was used to 3. Results determine significant differences (P < 0.05) for E. coli concentrations and each Recorded concentrations of E. coli antimicrobial resistance between sources. A varied greatly both within and between one-way ANOVA followed by a post hoc investigated rivers (Figure 2a). Highest E. coli concentrations were recorded in the Tukey’s HSD means test was used to -1 determine significant differences (P < 0.01) lower Brisbane River (3330 cfu 100 ml ) which also had the highest median E. coli between E. coli concentrations and each -1 antimicrobial between rivers. Spearman’s concentration (195 cfu 100 ml ) of all rank correlations (ρ < 0.05) were performed investigated rivers. The Maroochy and Pine

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Rivers shared the second highest E. coli > Mooloolah (16 cfu 100 ml-1) > Nerang (15 concentrations (95 cfu 100 ml-1) followed by cfu 100 ml-1) > Noosa (3 cfu 100 ml-1). A the Caboolture (55 cfu 100 ml-1) > Logan median concentration of 55 cfu 100 ml-1 was (48 cfu 100 ml-1) > Tweed (41 cfu 100 ml-1) recorded for the entire study.

3500 b 3000 A) 2500 2000 1500 b ab

1000 (cfu/100mls)

500 b E.. Coli Coli conentrationE.. 250 a ab ab ab 0 50 B) b b 40 b b 30 b 20 b

Ampicillin b % Resistant% 10 a 0 Max. 50 b C) 75th %ile 40 b c 30 b c median bc b 20 th

% Resistant% 25 %ile Tetracycline 10 a Min. 0 b 70 D) b 60 50 c b 40 30 bc c

% Resistant% 20 b

Sulfamethoxazole 10 a 0 10 E) b 8

6 b b

4 b

% Resistant% Ciprofloxacin 2 a a a a 0 Noosa Maroochy Mooloolah Caboolture Pine Lower Logan/ Nerang Brisbane Albert Figure 2: Comparison of South- East Queensland Rivers for A) average E. coli concentrations; B) % Ampicillin resistance; C) % Tetracycline resistance; D) % Sulfamethoxazole resistance; and E) % Ciprofloxacin resistance. Small different letters indicate significant differences (p<0.05) between rivers for each treatment.

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Ampicillin resistance in E. coli was was observed with tetracycline (4%) widespread but typically low with median (Figure 2d). Highest median sulfametho- values for all investigated rivers below 15% xazole resistance was seen in the Maroochy (Figure 2b). The highest median value was River (19%) followed by the lower Brisbane observed in the Logan/Albert River (14%) > Logan/Albert > Pine > Caboolture > followed by the Mooloolah > lower Brisbane Mooloolah > Nerang > Noosa Rivers. > Nerang > Caboolture > Pine and Maximum site sulfamethoxazole resistance Maroochy > Noosa Rivers. The highest site was seen in the lower Brisbane River (67%) ampicillin resistance was recorded in the with elevated resistance also recorded at lower Brisbane River (48%) with similar sites within the Maroochy and Caboolture elevated values recorded across other sites in Rivers. the Mooloolah, Caboolture, Logan/Albert E. coli resistance to ciprofloxacin was and Nerang Rivers. very low with a median value of 0% overall Overall resistance to tetracycline was with only the lower Brisbane (1%), approximately half that seen with ampicillin Logan/Albert (2%) and Pine (1%) Rivers with a median value of 7% (Figure 2c). The demonstrating medians greater than 0% highest median value was recorded in the (Figure 2e). Maximum site resistance to Logan/Albert (16%) followed by the Pine > ciprofloxacin was observed in the Maroochy > lower Brisbane > Mooloolah > Logan/Albert River (8%). Caboolture > Nerang > Noosa Rivers. Point-source (PS) sites appeared to be Maximum tetracycline resistance was the primary source of both E. coli and recorded at a site in the antimicrobial resistance with significantly (45%). greater (p< 0.05) colony counts and percent Resistance to sulfamethoxazole in all resistance than non-point source sites (NPS) investigated rivers was nearly half of what (Figure 3).

700 35 * Non-point Source 600 Point Source * 30

500 25

400 * 20 *

300 15 Conc.(CFU/100mls)

200 10 E.coli

100 5 Antibiotic Av. Resistance(%) Av. Av. * 0 0 E. coli Ampicillin Tetracycline Sulfamethoxazole Ciprofloxacin Figure 3: Comparison of Escherichia coli concentrations and associated antimicrobial resistance between investigated point and non-point source sites. *values indicate significant differences (p < 0.05) between sources.

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Significant correlations (ρ < 0.05) between median riverine ciprofloxacin were identified for E. coli concentration and resistance and total river WWTP discharge. both river WWTP discharge and river No other relationships were significant catchment population (Table 2). Significant (ρ > 0.05). correlations (ρ < 0.05) were identified

Table 2: Spearman’s rank correlations for selected river parameters and E. coli concentration and antimicrobial resistance.

River WWTP Discharge Volume River Population Median River Parameter Spearman r r value Spearman r r value E.coli Concentration 0.95 0.001 0.76 0.03 Ampicillin Resistance 0.69 0.06 0.32 0.42 Tetracycline Resistance 0.61 0.11 0.26 0.53 Sulfamethoxazole Resistance 0.21 0.6 0.08 0.83 Ciprofloxacin Resistance 0.82 0.01 0.49 0.21

4. Discussion discharge. Further work would suggest It is well established in the literature wastewater treatment plants offer a that the presence of Escherichia coli in favourable environment for the transfer and environmental surface waters is associated exchange of genetic information and could with degraded water quality (49-51). This enhance the development of antimicrobial study supports the literature with elevated E. resistance (12, 55-58). Additionally, Mao et coli found in the more degraded catchments. al 2015 (59) demonstrated that bacteria For example, the lower Brisbane and surviving the treatment process (including Logan/Albert Rivers frequently demonstrate chlorination) have a greater abundance of poor water quality with elevated nutrients antimicrobial resistance genes (ARG) and and suspended sediments, and low dissolved suggest this may be due to the selective pressure of significant concentrations of oxygen (52) and also recorded the highest median E. coli concentrations in this study. antimicrobials through the treatment stream In contrast, the Noosa, Mooloolah and and Novo et al 2013 (60) showed that Nerang Rivers are frequently characterised resistance was positively correlated with the by excellent water quality (52) and exhibited occurrence of antimicrobial residues. There very low E. coli concentrations. While there is some evidence to suggest that ARG are many reasons for these differences in carrying E.coli have a higher number of water quality, such as varying flow and virulence genes and therefore may present a catchment land use, WWTP discharge has greater risk for re-association with the been implicated as a major contributor to human microflora (61). these degraded rivers (53). This study has There was no correlation between also indicated that WWTP discharge is volume of wastewater discharge and highly correlated with E. coli concentrations. prevalence of antimicrobial resistance for all Research has shown that antimicrobial antimicrobials investigated, with the resistance increases with the level of exception of ciprofloxacin. This would exposure of faecal bacteria from humans suggest that other factors are more of an influence on the rates of antimicrobial (54) which may further support the increase in resistance with proximity to WWTP resistance seen within investigated E. coli.

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This may include antimicrobial use and resistant to cheaper drugs (67). And lastly, concentration within different source due to their restricted use in agriculture as catchments or treatment and disinfection growth promoters, there is little opportunity practices at WWTPs. Reinthaler et al. 2003 for the development and spread of resistance (12) have previously proposed that from low-dose exposure to these wastewater treatment will select for more antimicrobials as is typically seen with other antimicrobial resistant organisms and that agricultural antimicrobials. This is effluent disinfection, while lowering the total highlighted by the development of bacterial count, has also been shown to vancomycin resistance in Enterococci increase the prevalence of antimicrobial- following the use of avoparcin as a growth resistant bacteria (62, 63) and not be an promoter (68). effective control for antimicrobial resistance E. coli resistance to ciprofloxacin was at typical doses (18, 64). Therefore, the only investigated resistance correlated variations in treatment and disinfection are with WWTP discharge volume. Previous likely to lead to variations in antimicrobial investigations have identified a strong resistance in discharged organisms. correlation between the presence of Quinolone resistance amongst E. coli quinolone antimicrobials, such as was also significantly lower (p < 0.05) ciprofloxacin, norfloxacin and enrofloxacin, compared to the resistance to other and their proximity to WWTP discharge (7). investigated antimicrobials. There are a The use of quinolone antimicrobials have number of possible explanations for this and recently been banned in agriculture in the difference is probably driven by a Australia with their application confined combination of these. Firstly, quinolone solely for human and domestic animal antimicrobials were only introduced into treatment (69), indicating a potential driver clinical practice in the early 1970s, and as for their confinement to WWTP discharge. the first synthetic antimicrobials, the While the level of resistance to ciprofloxacin development of resistance to these drugs is appeared low it was comparable to clinical mostly limited to spontaneous mutants that estimates in both Queensland (3.4%) and alter target protein or increase pump Australia (3.4%), which are very low expression (65) and the spread of this themselves on a global scale (70). resistance is relatively restricted within the It has been previously demonstrated bacterial community. In contrast resistance that sites receiving wastewater discharge to the β-lactams, originally derived from a (PS) typically have a higher incidence of mould of the genus Penicillium, is antimicrobial resistance amongst their moderated by highly specific catalytic bacterial populations than NPS sites (36, 39, enzymes that were probably present before 47, 71, 72) and this study is no exception. As human development of antimicrobials, either the majority of bacteria in wastewater is of in other roles (for instance, housekeeping faecal origin, is it highly likely that the genes regulating sugar kinases or acetyl- antimicrobial resistance seen in this study is transferases also confer aminoglycoside acquired either within the original host or resistance) or as part of microbial during wastewater treatment; not from competition for an ecological niche (66). within receiving environment. However the Secondly, due to the comparatively low potential role of the environment in the incidence of clinical resistance and the fact transfer, development and storage of their synthetic nature makes production antimicrobial resistance should not be relatively expensive, quinolones are ignored. Additionally, the selective pressure generally reserved for treatment of infections

Copyright 2017 KEI Journals. All Rights Reserved Page │8 Medical Research Archives, Vol. 5, Issue 9, September 2017 Occurrence of antimicrobial resistant Escherichia coli in waterways of southeast Queensland, Australia of antimicrobial residues present in these to the presence of clinically relevant ARG waters may help maintain or even further on the chromosome of environmental develop resistance (73). Regulation of these bacteria (77), this is significantly different to PS facilities does not currently consider the clinical role ARG play through rapid antimicrobials or the level of resistance in horizontal and mobile transfer on genetic discharged bacteria in effluent licensing. elements, and then how these elements may Recent advancements and flexibility in the be maintained, transferred or further licensing approach has allowed for developed in the environmental setting. significant innovation in the region to Czekalski et al 2015 (39) has demonstrated include voluntary market-based mechanisms the potential for freshwater lakes to preserve for nutrient management to provide a better ARG and that background levels of environmental and cost-effective outcome resistance may be influenced by low level for eutrophication (74). It will be important, anthropogenic activities. Further, studies however, in the future to ensure this have shown the similarity of genes encoding flexibility adequately considers the role for CTX-M, an extended spectrum β- these point sources may play in the lactamase found in clinical isolates, with antimicrobial dissemination of resistance, chromosomal β-lactamases from an how to mitigate this process and its link to environmental Kluyvera spp. (78, 79) and other aspects of the water cycle (e.g. quinolone resistance (qnrA) on the Irrigation and drinking water). chromosome of Shewanella algae (80). Recent studies have identified an Given the relatedness of many of these important environmental reservoir of environmental microbes to pathogenic antimicrobial-resistance determinants, individuals, the further study of this termed the antimicrobial resistome, in soil environmental resistome, and the micro-organisms (75). As the environment, mechanisms driving it, could hold the key to particularly the soil microbial community, is the development of new means to combat the primary source of the antimicrobials we antimicrobial resistance. develop, it is not surprising that amongst these communities strategies have evolved to 5. Conclusions resist their actions. Even antimicrobial The aquatic environment is becoming producers themselves harbor resistance of increasing interest as a sink for elements for self-protection that are often antimicrobial resistant bacteria due to the clustered in operons (76). D’Costa and regular release of our waste streams colleagues (76) recently screened 480 soil (sewage, aquaculture and agricultural) Streptomyces representatives against 21 carrying antimicrobial residues and antimicrobials (both natural and antimicrobial resistant bacteria. This study semisynthetic) and found that without has identified antimicrobial resistant E. coli exception, all strains demonstrated some in rivers of South-East Queensland with level of multi-resistance. Dantas et. al. 2008 resistance greatest at sites receiving (75) have even gone so far as to show that discharge from WWTPs. The evidence soil bacteria can utilize a number of presented in this study highlights the need antimicrobials as a carbon source. This may for investigations into the mechanism of have implications for the use of effluent to antimicrobial resistance of discharged E. coli irrigate pasture and crops. and the potential for this resistance to be While it is suggested that most ARG transferred and maintained to the native originated from environmental isolates, due bacterial population.

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A modern-day cholera pandemic in focus against antimicrobial resistance, we Ecuador affected an estimated 8000 people cannot lose sight of the importance of the and was highlighted by the presence of a strategic role of the environment if we are to multi-resistance strain of V. cholera, which win the war. had been recently identified in multiple areas around the globe (81). While initial concerns Acknowledgements around the use of antimicrobials in local shrimp farms and its potential for the The authors wish to acknowledge development of this resistance were technical assistance provided by the staff at unfounded, the emergence of multiple Queensland Health Forensic and Scientific antimicrobial resistance in a virulent Services. This project was supported through opportunistic pathogen found in the an ARC Linkage Grant (LP0453-708) and in environment highlights the potential risk of part by the Wastewater Program of the the presence of antimicrobial resistance in Cooperative Research Centre for Water the aquatic environment. The impact of these Quality and Treatment (Project number observations on public health and the 666003). The use of trade, firm, or brand potential for the emergence and spread of names in this paper is for identification antimicrobial resistance through the aquatic purposes only and does not constitute environment demands further investigation endorsement by the University of to maintain our vigilance against the spread Queensland, Queensland Bulk Water Supply of infectious disease. While the battles rage Authority, University of Maryland or on the clinical front as the justified primary Queensland Health.

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References

1. WHO. Global Action Plan on 9. Huijbers PMC, Blaak H, de Jong MCM, Antimicrobial Resistance. In: Graat EAM, Vandenbroucke-Grauls C, Organisation WH, editor. Geneva, Husman AMD. Role of the Environment Switzerland: WHO Document in the Transmission of Antimicrobial Production Services; 2015. p. 28. Resistance to Humans: A Review. Environmental Science & Technology. 2. CoA. National Antimicrobial Resistance Strategy 2015-2019. In: Australia Co, 2015;49(20):11993-2004. editor. Canberra ACT: Commonwealth 10. Kummerer K. Pharmaceuticals in the of Australia; 2015. p. 48. Environment. Berlin: Springer; 2004. 3. Batt AL, Bruce IB, Aga DS. Evaluating 527 p. the vulnerability of surface waters to 11. Marti E, Balcázar JL. Antibiotic antibiotic contamination from varying Resistance in the Aquatic Environment. wastewater treatment plant discharges. Comprehensive Analytical Chemistry. Environ Pollut. 2006;142(2):295-302. 2013;62:671-84. 4. Calamari D, Zuccato E, Castiglioni S, 12. Reinthaler FF, Posch J, Feierl G, Wust Bagnati R, Fanelli R. Strategic survey of G, Haas D, Ruckenbauer G, et al. therapeutic drugs in the rivers Po and Antibiotic resistance of E. coli in Lambro in northern Italy. sewage and sludge. Water Research. Environmental Science and Technology. 2003;37(8):1685-90. 2003;37(7):1241-8. 13. Watkinson AJ, Murby EJ, Costanzo SD. 5. Costanzo SD, Murby J, Bates J. Removal of antibiotics in conventional Ecosystem response to antibiotics and advanced wastewater treatment: entering the aquatic environment. Implications for environmental Marine Pollution Bulletin. 2005;51:218- discharge and wastewater recycling. 23. Water Research. 2007;doi:10.1016/j.watres.2007.04.005. 6. Kolpin DW, Furlong ET, Meyer MT, Thurman EM, Zaugg SD, Barber LB, et 14. Guardabassi L, Petersen A, Olsen JE, al. Pharmaceuticals, hormones, and Dalsgaard A. Antibiotic Resistance in other organic wastewater contaminants Acinetobacter spp. Isolated from Sewers in US streams, 1999-2000: A national Receiving Waste Effluent from a reconnaissance. Environmental Science Hospital and a Pharmaceutical Plant. & Technology. 2002;36(6):1202-11. Appl Environ Microbiol. 7. Watkinson AJ, Murby EJ, Kolpin DW, 1998;64(9):3499-502. Costanzo SD. The occurrence of 15. Gallert C, Fund K, Winter J. Antibiotic antibiotics in an urban watershed:from resistance of bacteria in raw and wastewater to drinking water. Science of biologically treated sewage and in the Total Environment. 2009;407:2711- groundwater below leaking sewers. 23. Applied Microbiology and 8. Carvalho IT, Santos L. Antibiotics in the Biotechnology. 2005;69(1):106-12. aquatic environments: A review of the 16. Martins da Costa P, Vaz-Pires P, European scenario. Environment Bernardo F. Antimicrobial resistance in International. 2016;94:736-57. Enterococcus spp. isolated in inflow,

Copyright 2017 KEI Journals. All Rights Reserved Page │11 Medical Research Archives, Vol. 5, Issue 9, September 2017 Occurrence of antimicrobial resistant Escherichia coli in waterways of southeast Queensland, Australia

effluent and sludge from municipal Australia. Journal of Applied sewage water treatment plants. Water Microbiology. 2006;100(5):1103-13. Research. 2006;40(8):1735-40. 24. Le TX, Munekage Y, Kato S-i. 17. Ziembinska-Buczynska A, Felis E, Antibiotic resistance in bacteria from Folkert J, Meresta A, Stawicka D, Gnida shrimp farming in areas. A, et al. Detection of antibiotic Science of The Total Environment. resistance genes in wastewater treatment 2005;349(1-3):95-105. plant - molecular and classical approach. 25. Jordan D, Morris SG, Gill P, Andersen Archives of Environmental Protection. LM, Chowdhury A, Stevenson AE, et al. 2015;41(4):23-32. Mass screening for antimicrobial 18. Yuan QB, Guo MT, Yang J. Fate of resistant Escherichia coli in dairy cows Antibiotic Resistant Bacteria and Genes in northern New South Wales. during Wastewater Chlorination: Australian Veterinary Journal. Implication for Antibiotic Resistance 2005;83(11):688-94. Control. Plos One. 2015;10(3). 26. de Fatima Silva Lopes M, Ribeiro T, 19. Bjorklund H, Bondestam J, Bylund G. Abrantes M, Figueiredo Marques JJ, Residues of oxytetracycline in wild fish Tenreiro R, Crespo MTB. Antimicrobial and sediments from fish farms. resistance profiles of dairy and clinical Aquaculture. 1990;86(4):359-68. isolates and type strains of enterococci. 20. Chelossi E, Vezzulli L, Milano A, International Journal of Food Branzoni M, Fabiano M, Riccardi G, et Microbiology. 2005;103(2):191-8. al. Antibiotic resistance of benthic 27. Chee-Sanford JC, Aminov RI, Krapac bacteria in fish-farm and control IJ, Garrigues-Jeanjean N, Mackie RI. sediments of the Western Occurrence and Diversity of Mediterranean. Aquaculture. Tetracycline Resistance Genes in 2003;219(1-4):83-97. Lagoons and Groundwater Underlying 21. Guardabassi L, Dalsgaard A, Raffatellu Two Swine Production Facilities. Appl Environ Microbiol. 2001;67(4):1494- M, Olsen JE. Increase in the prevalence of oxolinic acid resistant Acinetobacter 502. spp. observed in a stream receiving the 28. Mathew AG, Arnett DB, Cullen P, effluent from a freshwater trout farm Ebner PD. Characterisation of resistance following the treatment with oxolinic patterns and detection of apramycin acid-medicated feed. Aquaculture. resistance genes in Escherichia coli 2000;188(3-4):205-18. isolated from swine exposed to various 22. Molina AA, Garcia GA, Abreu GA, environmental conditions. International Bolan MC, Roque A, Gomez GB. Journal of Food Microbiology. Plasmid profiling and antibiotic 2003;89:11-20. resistance of Vibrio strains isolated from 29. Sengelov G, Agerso Y, Halling- cultured penaeid shrimp. FEMS Sorensen B, Baloda SB, Andersen JS, Microbiology Letters. 2002;213(1):7-12. Jensen LB. Bacterial antibiotic 23. Akinbowale OL, Peng H, Barton MD. resistance levels in Danish farmland as a Antimicrobial resistance in bacteria result of treatment with pig manure isolated from aquaculture sources in slurry. Environment International. 2003;28(7):587-95.

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30. Obeng AS, Rickard H, Sexton M, Pang 37. Rosas I, Salinas E, Martinez L, Cruz- Y, Peng H, Barton M. Antimicrobial Cordova A, Gonzalez-Pedrajo B, susceptibilities and resistance genes in Espinosa N, et al. Characterization of Campylobacter strains isolated from Escherichia coli Isolates from an Urban poultry and pigs in Australia. Journal of Lake Receiving Water from a Applied Microbiology. Wastewater Treatment Plant in Mexico 2012;113(2):294-307. City: Fecal Pollution and Antibiotic Resistance. Current Microbiology. 31. Obeng AS, Rickard H, Ndi O, Sexton M, Barton M. Antibiotic resistance, 2015;71(4):490-5. phylogenetic grouping and virulence 38. Rathnayake I, Hargreaves M, Huygens potential of Escherichia coli isolated F. SNP diversity of Enterococcus from the faeces of intensively farmed faecalis and Enterococcus faecium in a and free range poultry. Veterinary South East Queensland waterway, Microbiology. 2012;154(3-4):305-15. Australia, and associated antibiotic 32. Ash RJ, Mauck B, Morgan M. resistance gene profiles. Bmc Antibiotic Resistance of gram-negative Microbiology. 2011;11. bacteria in Rivers, United States. 39. Czekalski N, Sigdel R, Birtel J, Emerging Infectious Diseases. Matthews B, Burgmann H. Does human 2002;8(7):713-6. activity impact the natural antibiotic 33. Goni-Urriza M, Capdepuy M, Arpin C, resistance background? Abundance of Raymond N, Caumette P, Quentin C. antibiotic resistance genes in 21 Swiss Impact of an Urban Effluent on lakes. Environment International. Antibiotic Resistance of Riverine 2015;81:45-55. Enterobacteriaceae and Aeromonas spp. 40. Barker-Reid F, Fox EM, Faggian R. Appl Environ Microbiol. Occurrence of antibiotic resistance 2000;66(1):125-32. genes in reclaimed water and river water 34. Harakeh S, Yassine H, El-Fadel M. in the Werribee Basin, Australia. Antimicrobial-resistant patterns of Journal of Water and Health. Escherichia coli and Salmonella strains 2010;8(3):521-31. in the aquatic Lebanese environments. 41. Enne VI, Livermore DM, Stephens P, Environ Pollut. 2006;143(2):269-77. Hall LMC. Persistence of sulphonamide 35. Miranda CD, Zemelman R. Antibiotic resistance in Escherichia coli in the UK Resistant Bacteria in Fish from the despite national prescribing restriction. Concepcion Bay, Chile. Marine Lancet. 2001;357(9265):1325-8. Pollution Bulletin. 2001;42(11):1096- 42. Mammeri H, Van de Loo M, Poirel L, 102. Martinez-Martinez L, Nordmann P. 36. Parveen S, Murphree R, Edmiston L, Emergence of plasmid-mediated Kaspar C, Portier K, Tamplin M. quinolone resistance in Escherichia coli Association of multiple-antibiotic- in Europe. Antimicrobial Agents and resistance profiles with point and Chemotherapy. 2005;49(1):71-6. nonpoint sources of Escherichia coli in 43. Galland JC, Hyatt DR, Crupper SS, Apalachicola Bay. Applied and Acheson DW. Prevalence, Antibiotic Environmental Microbiology. Susceptibility, and Diversity of 1997;63(7):2607-12. Escherichia coli O157:H7 Isolates from a Longitudinal Study of Beef Cattle

Copyright 2017 KEI Journals. All Rights Reserved Page │13 Medical Research Archives, Vol. 5, Issue 9, September 2017 Occurrence of antimicrobial resistant Escherichia coli in waterways of southeast Queensland, Australia

Feedlots. Appl Environ Microbiol. total coliform, fecal coliform, and 2001;67(4):1619-27. enterococcus bacterial indicator 44. Lanz R, Kuhnert P, Boerlin P. response for ocean recreational water Antimicrobial resistance and resistance quality testing. Water Research. gene determinants in clinical 2003;37:1637-43. Escherichia coli from different animal 52. EHMP. Ecosytem Health Monitoring species in Switzerland. Veterinary Program: 2005-06 Annual Technical Microbiology. 2003;91(1):73-84. Report. Brisbane, Australia: South East 45. Scott L, McGee P, Minihan D, Sheridan Queensland Healthy Waterways JJ, Earley B, Leonard N. The Partnership; 2007. characterisation of E. coli O157:H7 53. Dennison WC, Abal EG. Moreton Bay isolates from cattle faeces and feedlot Study: A scientific Basis for the Healthy environment using PFGE. Veterinary Waterways Campaign. Brisbane, Microbiology. 2006;114(3-4):331-6. Queensland: South East Queensland 46. Bucknell DG, Gasser RB, Irving A, Regional Water Quality Management Whithear K. Antimicrobial resistance in Strategy; 1999. Salmonella and Escherichia coli isolated 54. Skurnik D, Ruimy R, Andremont A, from horses. . Australian Veterinary Amorin C, Rouquet P, Picard B, et al. Journal. 1997; 75:355-6. Effect of human vicinity on 47. Watkinson AJ, Micalizzi GB, Bates JB, antimicrobial resistance and integrons in Costanzo SD. A novel method for rapid animal faecal Escherichia coli. J assessment of antibiotic resistance in Antimicrob Chemother. 2006;57(6):1215-9. Escherichia coli from environmental waters using a modified Chromogenic 55. Inoue D, Sei K, Soda S, Ike M, Fujita agar. Applied and Environmental M. Potential of predominant activated Microbiology. 2007;73(7). sludge bacteria as recipients in 48. Giger W, Alder AC, Golet EM, Kohler conjugative plasmid transfer. Journal of Bioscience and Bioengineering. HPE, McArdell CS, Molnar E, et al. Occurrence and fate of antibiotics as 2005;100(6):600-5. trace contaminants in wastewaters, 56. Mach PA, Grimes DJ. R-plasmid sewage sludges, and surface waters. transfer in a wastewater treatment plant. Chimia. 2003;57(9):485-91. Applied and Environmental Microbiology. 1982;44(6):1395-403. 49. Davies M, Long J, Donald M, Ashbolt N. Survival of Fecal Microorganisms in 57. Mezrioui N, Baleux B. Resistance Marine and Freshwater Sediments. Patterns of Escherichia-Coli Strains Applied and Environmental Isolated from Domestic Sewage before Microbiology. 1994;61:1888-96. and after Treatment in Both Aerobic Lagoon and Activated-Sludge. Water 50. Mallin M, Williams K, Esham E, Lowe R. Effect of Human Development on Research. 1994;28(11):2399-406. bacteriological water quality in coastal 58. Czekalski N, Berthold T, Caucci S, Egli watersheds. Ecological Applications. A, Buergmann H. Increased levels of 2000;10:1047-56. multiresistant bacteria and resistance genes after wastewater treatment and 51. Noble R, Moore D, Leecaster M, McGee C, Weisberg S. Comparison of their dissemination into Lake Geneva,

Copyright 2017 KEI Journals. All Rights Reserved Page │14 Medical Research Archives, Vol. 5, Issue 9, September 2017 Occurrence of antimicrobial resistant Escherichia coli in waterways of southeast Queensland, Australia

Switzerland. Frontiers in Microbiology. 66. Baquero F, Blazquez J. Evolution of 2012;3(106):1-18. antibiotic resistance. Trends in Ecology 59. Mao DQ, Yu S, Rysz M, Luo Y, Yang & Evolution. 1997;12(12):482-7. FX, Li FX, et al. Prevalence and 67. TGL. Theraputic Guidelines: Antibiotic proliferation of antibiotic resistance Version 12. North Melbourne, Australia: genes in two municipal wastewater Therapeutic Guidelines Limited; 2003. treatment plants. Water Research. 68. Collignon P. Vancomycin-resistant 2015;85:458-66. enterococci and use of avoparcin in 60. Novo A, Andre S, Viana P, Nunes OC, animal feed: is there a link? Medical Manaia CM. Antibiotic resistance, Journal of Australia. 1999;171:144-6. antimicrobial residues and bacterial 69. Unicomb LE, Ferguson J, Stafford RJ, community composition in urban Ashbolt R, Kirk MD, Becker NG, et al. wastewater. Water Research. Low-Level Fluoroquinolone Resistance 2013;47(5):1875-87. among Campylobacter jejuni Isolates in 61. Biswal BK, Mazza A, Masson L, Gehr Australia. Clin Infect Dis. R, Frigon D. Impact of wastewater 2006;42(10):1368-74. treatment processes on antimicrobial 70. Pearson J, Turnidge J, Franklin C, Bell resistance genes and their co-occurrence J. Prevalence of Antimicrobial with virulence genes in Escherichia coli. Resistances in Common Pathogenic Water Research. 2014;50:245-53. Enterobacteriaceae in Australia, 2004: 62. Murray GE, Tobin RS, Junkins B, Report from the Australian Group on Kushner DJ. Effect of Chlorination on Antimicrobial Resistance. Antibiotic-Resistance Profiles of Communicable Diseases Intelligence. Sewage-Related Bacteria. Applied and 2007;31:106-12. Environmental Microbiology. 71. Boon PI, Cattanach M. Antibiotic 1984;48(1):73-7. resistance of native and faecal bacteria 63. Watkinson AJ, Micalizzi GB, Graham isolated from rivers, reservoirs and GM, Bates JB, Costanzo SD. Antibiotic- sewage treatment facilities in Victoria, Resistant Escherichia coli in south-eastern Australia. Lett Appl Wastewaters, Surface Waters, and Microbiol. 1999;28(3):164-8. Oysters from an Urban Riverine System. 72. Koczura R, Mokracka J, Jablonska L, Applied and Environmental Gozdecka E, Kubek M, Kaznowski A. Microbiology. 2007;73(17):1-2. Antimicrobial resistance of integron- 64. Zhang YY, Zhuang Y, Geng JJ, Ren harboring Escherichia coli isolates from HQ, Zhang Y, Ding LL, et al. clinical samples, wastewater treatment Inactivation of antibiotic resistance plant and river water. Science of the genes in municipal wastewater effluent Total Environment. 2012;414:680-5. by chlorination and sequential 73. Tello A, Austin B, Telfer TC. Selective UV/chlorination disinfection. Science of pressure of antibiotic pollution on the Total Environment. 2015;512:125- bacteria of importance to public health. 32. Environmental Health Perspectives. 65. Hooper DC. Fluoroquinolone resistance 2012;128(8):1100-6. among Gram-positive cocci. The Lancet 74. DEHP. Flexible options for managing Infectious Diseases. 2002;2(9):530-8. point source water emissions: A

Copyright 2017 KEI Journals. All Rights Reserved Page │15 Medical Research Archives, Vol. 5, Issue 9, September 2017 Occurrence of antimicrobial resistant Escherichia coli in waterways of southeast Queensland, Australia

voluntary market-based mechanism for Lactamases Antimicrobial Agents and nutrient management. In: Protection Chemotherapy. 2002;46:4038-40. DoEaH, editor. Brisbane, Australia: 79. Humeniuk C, Arlet G, Gautier V, State of Queensland; 2014. p. 9. Grimont P, Labia R, Philippon A. b- 75. Dantas G, Sommer MOA, Oluwasegun Lactamases of Kluyvera ascorbata, RD, Church GM. Bacteria subsisting on probable progenitors of some plasmid- antibiotics. Science. 2008;320:100-3. encoded CTX-M types. Antimicrobial 76. D'Costa VM, McGrann KM, Hughes Agents and Chemotherapy. DW, Wright GD. Sampling the 2002;46:3045–9. antibiotic resistome. Science. 80. Poirel L, Rodriguez-Martinez J, 2006;311:374-7. Mammeri H, Liard A, Nordmann P. 77. Cantas L, Shah SQA, Cavaco LM, Origin of Plasmid-Mediated Quinolone Manaia CM, Walsh F, Popowska M, et Resistance Determinant QnrA al. A brief multi-disciplinary review on Antimicrobial Agents and antimicrobial resistance in medicine and Chemotherapy. 2005;49:3523-5. its linkage to the global environmental 81. Smith P. Antimicrobial use in shrimp microbiota. Frontiers in Microbiology. farming in Ecuador and emerging multi- 2013;4. resistance during the cholera epidemic 78. Poirel L, Kampfer P, Nordmann P. of 1991: A re-examination of the data. Chromosome-Encoded Ambler Class A Aquaculture. 2007;271(1-4):1-7. β-Lactamase of Kluyvera georgiana, a 82. SEQRWMS. Discover the waterways of Probable Progenitor of a Subgroup of South-east Queensland. Brisbane: South CTX-M Extended-Spectrum β- East Queensland Regional Wastewater Management Strategy; 2001.

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