Framework for monitoring antibiotic content and antibiotic resistance in the Danube Delta - the EnviroAMR project -

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1 Framework for monitoring antibiotic content and antibiotic resistance in the Danube Delta - the EnviroAMR project - Dr. Cristian COMAN Institute of Biological Research Cluj-Napoca

2 November 18 th November 16-22

3 The problem of AMR in the environment. The EnviroAMR goals. Plan in the Danube Delta.

4 Antibiotics Antibiotic - A medicine (such as penicillin or its derivatives) that inhibits the growth of or destroys microorganisms (Oxford dictionary)

5 No. of antibiotics approved for human usage in the past 30 years (Source: Cole, 2014)

6 Class of antibiotic Representative drug Mode of action β Lactam Sulfonamides Aminoglycosides Glycopeptides Lincosamides Macrolides Rifamycins Tetracyclines Chloramphenicol Quinolones Fluoroquinolones Phenoxypenicillin, oxacillin, amoxicillin, carbenicillin, piperacillin Sulfamethoxazole, sulfamethazine, Sulfdiazine, sulfadimethoxine, Sulfamethoxazole, Sulfasalazine, sulfasoxazole etc. Gentamycin Vancomycin Clindamycin, lincomycin Clarithromycin, erythromycin, Spiramycin, tylosin Rifamycin Chlortetracycline,Doxycycline, oxytetracycline, Tetracycline etc. Chloramphenicol Oxolinic acid, nalidixic acid, pipemidic acid, flumequine, pefloxacin Ciprofloxacin, norfloxacin, ofloxacin, enrofloxacin, enoxacin, sarafloxacin etc. Inhibits steps in cell wall synthesis and murein assembly Inhibitors of bacterial beta lactamases Inhibits translation (protein synthesis) Inhibits steps in murein biosynthesis and assembly Inhibits translation (protein synthesis) Inhibits translation (protein synthesis) Inhibits transcription (bacterial RNA polymerase) Inhibits translation (protein synthesis) Inhibits translation (protein synthesis) Inhibits DNA replication Inhibits DNA replication (After Gothwal and Shashidhar, 2014)

7 Antimicrobial resistance - resistance of a microorganism to an antimicrobial drug that was originally effective for treatment of infections caused by it. (World Health Organization)

8 Antimicrobial resistance Intrinsic Acquired Mutation Transffered Transformation Transduction Conjugation

9 Is AMR that important? [ ] there is the danger that the ignorant man may easily underdose himself and by exposing his microbes to non-lethal quantities of the drug make them resistant. (A. Fleming, Nobel lecture, 1945) [ ] arguably the greatest risk to human health comes in the form of antibiotic-resistant bacteria. (World Economic Forum, 2013) We acknowledge that the emergence of AMR is an increasing global health threat and that the spread of AMR affects all nations. (Berlin Declaration on Antimicrobial Resistance, G7 Health Ministers, October 2015)

10 (Source: Jim O Neill The Review on AMR, 2015) (Front cover The Economist, May 21 st 2016)

11 Together with other 18 EU countries, Romania is part of the Joint Programming Initiative on Antimicrobial Resistance (JPI AMR) (JPI AMR Strategic Research Agenda, 2013) October 2015 JPI AMR workshop in Berlin -> what to focus on in the near future?

12 Why focus on the environment? Antibiotics, ARBs and ARGs new classes of pollutants in the environment (Source: Coutinho et al. 2013)

13 Why focus on the environment?

14 In Romania there is not a comprehensive system for monitoring antibiotics and AMR in the environment => the EnviroAMR project

15 The EnviroAMR project Financed through the EEA Financial Mechanism, in the RO04 Reduction of hazardous substances. Will raise awareness that antibiotics and antimicrobial resistant microorganisms are considered novel classes of pollutants in the environment. Programme Operator: Ministry of Environment, Waters and Forests Will propose a methodological guide for standardized monitoring of antibiotics and antibiotic resistance in the environment. Start date: May 20 th 2015 End date: October 31 st 2016 The beneficiaries are public entities and stakeholders directly involved in implementing strategies for a clean and healthy environment.

16 Raising awareness is very important! ARG WWTP inlet WWTP outlet River downstream of WWTP Hospital sewage Subsurface water Pig manure erma sul sul teta tetb tetc teto tetw (Source: EnviroAMR)

17 The EnviroAMR project a methodological guide Environmental sampling for monitoring. I. SAMPLING Detection and cuantification of antibiotics. II. CHEMICAL ANALYSIS Data integration and analysis. VI. INTEGRATE Antimicrobial susceptibility testing. Deposit the AMR strains in a biobank. III. PHENOTYPE Metagenomics (metabarcoding and shotgun): from sample preparation to bioinformatics. V. METAGENOME Determine the 16S rrna gene and ARG sequences. qpcr of target ARGs. IV. GENOTYPE

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20 JDS3 report (www. icpdr.org)

21 Danube Delta expedition Research collaboration agreement between the Institute of Biological Research Cluj-Napoca and the National Institute for R&D Danube Delta Tulcea signed in May 2016 Direct collaboration with Dr. Liliana Torok (Toxicology lab) Molecular investigation of cyanobacterial blooms and cyanotoxins in the Danube Delta. Can cyanobacterial blooms trigger/influence the natural AMR?

22 Danube Delta expedition (Source: RIZA and DDNI rapport )

23 Danube Delta expedition Băclănești Furtuna Nebunu Merhei Matița Dracului Miazăzi Trei Iezere Isac Uzlina Cuibul cu Lebede Gorgoștel Roșu Roșuleț Puiu Iacob Erenciuc

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