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Antibiotic Resistance in Gram Negative Bacteria Isolated from Fish Sold in Western Australia

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Antibiotic Resistance in Gram Negative Bacteria Isolated from Fish Sold in Western Australia Edith Cowan University Research Online Theses: Doctorates and Masters Theses 2018 Antibiotic resistance in Gram negative bacteria isolated from fish sold in Western Australia Hannah Kathleen Robinson Edith Cowan University Follow this and additional works at: https://ro.ecu.edu.au/theses Part of the Food Science Commons, Nutrition Commons, and the Public Health Commons Recommended Citation Robinson, H. K. (2018). Antibiotic resistance in Gram negative bacteria isolated from fish sold in esternW Australia. https://ro.ecu.edu.au/theses/2161 This Thesis is posted at Research Online. https://ro.ecu.edu.au/theses/2161 Edith Cowan University Copyright Warning You may print or download ONE copy of this document for the purpose of your own research or study. The University does not authorize you to copy, communicate or otherwise make available electronically to any other person any copyright material contained on this site. You are reminded of the following: Copyright owners are entitled to take legal action against persons who infringe their copyright. A reproduction of material that is protected by copyright may be a copyright infringement. Where the reproduction of such material is done without attribution of authorship, with false attribution of authorship or the authorship is treated in a derogatory manner, this may be a breach of the author’s moral rights contained in Part IX of the Copyright Act 1968 (Cth). Courts have the power to impose a wide range of civil and criminal sanctions for infringement of copyright, infringement of moral rights and other offences under the Copyright Act 1968 (Cth). Higher penalties may apply, and higher damages may be awarded, for offences and infringements involving the conversion of material into digital or electronic form. Antibiotic resistance in Gram negative bacteria isolated from fish sold in Western Australia This thesis is presented in fulfilment of the degree of the Master of Public Health Hannah Kathleen Robinson Edith Cowan University School of Medical and Health Sciences 2018 Abstract Background Information The global misuse and overuse of antibiotics in human medicine and the animal production industry is contributing to the development of antibiotic resistance in bacteria. This is a serious threat to modern medicine and public health. Antibiotic resistant organisms can cause severe infections in humans which are difficult to treat, and in some cases impossible to resolve which can lead to premature death. Several studies have been conducted across the globe to assess the use of antibiotics in the seafood industry and the associated health risks, however, limited studies have recently explored this risk in an Australian setting. Aims This thesis aimed to investigate the presence of antibiotic residues in seafood sold in Western Australia. Furthermore, the occurrence of antibiotic resistance in Gram negative bacteria isolated from fish sold in Perth, Western Australia was assessed. The impact of country of origin on the presence of antibiotic residues and antibiotic resistant bacteria in seafood samples has also been considered. Methodology Historical data was accessed from the Local Health Authorities Analytical Committee regarding the presence of eight antibiotic types in 253 seafood samples purchased throughout Western Australia between May and June 2017. Forty-four fish samples, a mix of local and imported from Asian countries, were sourced from retail shops located in the metropolitan area of Perth between September and November 2017. Gram negative bacteria were isolated by homogenisation of the fish with a Luria Bertani Broth and incubation on media selective for Gram negative bacteria. A series of preliminary microbial identification tests were conducted on selected bacterial isolates. Matrix Assisted Laser Desorption Ionization-Time of Flight confirmed the identification of the bacteria to species level. The identified bacteria (n = 35) were analysed for antibiotic susceptibility to eight antibiotic types using the standard disc diffusion method. Results The majority of seafood samples were free from antibiotic residue contamination and compliant with Australian legislation. A single non-compliant sample contained antibiotic residues below the level required to pose an immediate health risk to the consumer. This ii result suggests the Australian consumer has limited risk of consuming antibiotic residues in seafood. Thirty-five Gram negative bacterial isolates from ten genera were identified. The majority of the antibiotic resistance observed in the bacteria was either explained by intrinsic resistance or was similar to previous reports. Potential acquired antibiotic resistance was observed in four Acinetobacter species and a Rhizobium isolate which were isolated from commonly farmed fish from Australia (n = 1), China (n = 1) and Vietnam (n = 3). It is possible the fish may have been exposed to antibiotics during the production cycle. However, this result must be read with caution since there are limited standardised breakpoint guidelines for these particular species and, therefore the results were inferred using guidelines for other, similar, bacterial species. From these results, it appears that there is limited risk to consumer health from exposure to antibiotic resistant bacteria via consumption of seafood, however, only a limited number of samples were assessed, and Gram positive bacteria were not evaluated in this study. These results are reassuring but suggest that vigilance is required to ensure that the risk to consumers is minimised. Where antibiotics are used inappropriately in environmental settings, the risk of environmental bacteria developing further antibiotic resistance will remain. Routine surveillance of antibiotic resistance in pathogenic bacteria in domestic and imported food of animal origin is recommended to monitor this potential risk to human public health. iii Declaration I certify that this thesis does not, to the best of my knowledge and belief: i. incorporate without acknowledgment any material previously submitted for a degree or diploma in any institution of higher education; ii. contain any material previously published or written by another person except where due reference is made in the text of this thesis; or iii. contain any defamatory material; Hannah Kathleen Robinson 21 December 2018 iv Acknowledgements I would like to express my gratitude and appreciation to my supervisors, Dr Megan Lloyd, Ms Kerry Staples and Associate Professor Jacques Oosthuizen. Thank you for your guidance and patience. Thank you also to Mr Trevor Chapman of the Local Health Authorities Analytical Committee for supporting this research from the beginning. I am grateful for the kind permission to access your coordinated sampling project data. I would also like to thank Professor Tom Riley and the Clostridium difficile research staff for their generous assistance in providing microbial identification and for their analysis of the fish samples for the presence of Clostridium difficile. The analysis of the fish in this study for Clostridium difficile was conducted with funding provided by the School of Medical and Health Sciences Intra-School Collaboration Seed Fund, which is a funding initiative of Edith Cowan University. Finally, I would like to thank my family and friends for their unfailing encouragement. Special thanks to my Mum, Dad, Jeffrey, Jane, Leanne and Sarah. v Table of Contents Abstract .................................................................................................................................................................. ii Declaration ............................................................................................................................................................ iv Acknowledgements ................................................................................................................................................ v Table of Contents .................................................................................................................................................. vi Abbreviations ....................................................................................................................................................... viii Naming Conventions ........................................................................................................................................... viii Glossary ................................................................................................................................................................. ix 1.0 Introduction ...................................................................................................................................................... 1 2.0 Literature Review .............................................................................................................................................. 2 2.1 Antibiotic Resistance .................................................................................................................................... 2 2.1.1 Mechanisms of Antibiotic Resistance ................................................................................................... 2 2.1.2 Health and Economic Impacts of Antibiotic Resistance ........................................................................ 2 2.1.3 Development of New Antibiotics .........................................................................................................
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