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1 Fate of antibiotics and antibiotic-resistant fecal bacteria from the contamination source to the estuary: impact and/or resilience? French-Japanese conference.

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Presentation on theme: "1 Fate of antibiotics and antibiotic-resistant fecal bacteria from the contamination source to the estuary: impact and/or resilience? French-Japanese conference."— Presentation transcript:

1 1 Fate of antibiotics and antibiotic-resistant fecal bacteria from the contamination source to the estuary: impact and/or resilience? French-Japanese conference of oceanography Boulogne sur Mer 2013 Fabienne Petit UMR CNRS 6143

2 2 Health context:  Emergence and reemergence of infectious diseases Shiga-toxin E.coli (STEC)  Emergence of antibiotic-resistant bacteria Environmental context:  Increase of anthropic pressure on the watershed: population, industrial activities, farming practices  Higher vulnerability of the estuaries and the coastal zones to contamination, by pathogens and pharmaceutical contaminants  Increase of the risk for human health via shellfish consumption;  Existence of a putative risk related to the persistence and the spread of the virulence and antibiotic-resistant genes. Scientific and societal contexts A challenge for the next decade: assessment of how ecosystem changes could affect human health © Manfred Rohde/Helmholtz-Zentrum für Infektions/AFP Escherichia coli O104 Millenium ecosystem assessment, Who

3 to tonnes per year (Wise et al., 2002) (Affsaps, 2011) (who, 2001)  The increase of antibiotic resistance is a major public healthissue worldwide. Scientific and societal contexts  France: the world’s 4th largest consumer of drugs  The Seine estuary is the receiving water body from a highly anthropized catchment area: 79,000km 2 30% of the French population lives along the estuary 40% of industrial and agricultural activities are located on the watershed Haute-Normandie : one of the highest antibiotic consumer

4 4 Antibiotic use and overuse Selection of antibio-resistant Escherichia coli, and Enterococcus Soil leaching, fields run off septic tank WWTP Human exposure Is there a transfer of antibiotic-resistance or virulent gene? What about the integron spread which plays a keyrole in multiple antibiotic- resistance? Is antibiotic concentration sufficient for the emergence of antibiotic-resistant bacteria ? Sediments Mussels Water Gene transfer Release of antibiotics and antibio-resistant bacteria Antibiotic persistence Scientific and societal contexts Why does the ecosystem matter for human health in the emergence of antibiotic resistance?

5 5  Antibiotic contamination : from 40 ng.L -1 (Togola, 08) to 100 ng.L-1 in water (Tamtam et al, 09). Antibiotic and antibiotic-resistant bacteria in the Seine estuary 654 strains Laroche et al., 09  A permanent high level of antibiotic-resistant E. coli (from 30 to 56% among which 11% contains integrons)  Low level of microbial quality throughout the Seine estuary due to: the upstream input of WWTP; the release of high contaminated tributaries Touron et al., 07 Percentage of antibiotic-resistant E. coli 6.5millions inhabitants eq. The main conclusions of the previous scientific programs ( )

6 6 Percentage of samples of low microbial quality (E. coli > 1000 CFU.100 g -1 CLI) Honfleur [5-25%][25-50%] Putative human exposure: microbial quality of shellfish Programme seine aval Ifremer data  Contamination of the mussels from coastal zone depending on the sampling site  Occurrence of antibiotic-resistant E. coli in mussels sampled in the mouth of the estuary (Honfleur)

7 7 The multidisciplinary FLASH project: EC2CO Based on those results, the main objectives of the FLASH project are to have a better understanding of :  the fate of antibiotic, and of the antibiotic-resistant bacteria (E.coli, Enterococcus) and the corresponding genes;  the relationship between antibiotic and antibiotic-resistant bacteria in water;  the identification of the source of the contamination by antibiotic-resistant bacteria (human or bovine). Two water continuum were monitored during two years (6 sampling campaigns):  A rural land use continuum;  A medical center - wastewater treatment plant - river continuum

8 8  the most frequently antibiotic prescribed ( Penicillin/amoxicillin) was detected in the water in low concentration  The most persistent antibiotic were detected : quinolones, cephalosporin, macrolides  The antibiotic concentration is too low to exert a selective pressure on bacteria Antibiotic contamination in water from medical center to river Antibiotic use in medical center Development of a new chemical methodology to detect 34 antibiotics ( nG.L -1 ) LPTC Antibiotic concentration in hospital and retirement home effluents

9 9 Antibioresistant E.coli (%, N=50) Relationship between antibiotic use, antibiotic contamination, and antibiotic resistant bacteria, in medical center effluents  Antibiotic-resistance is linked to an antibiotic prescription  no similar observation for both tetracyclin and sulfonamids resistance : Integrons? Antibio- resistant Enteroccocus( %, N=50) Antibiotic consumption during high epidemic period ( december 2009) One month before the sampling hospital ( 87 beds, stay of short time) retirement home (180beds, average stay of 10 years)

10 10 Resistant to at least 3 antibiotics Harboring class 1 integron  E.coli resistant to at least 3 antibiotics and those carrying class1 integron decrease significantly ( Oberlé et al., 2012)  Lower survival of antibiotic-multi-resistant E.coli strains, which were replaced by strains better adapted to the water environment (Berthe et al, 2013) Occurence of multiple antibiotic resistance and class 1 integron in waters Along the medical center-wastewater treatment plant- river continuum (4km) * (p-value<0,001) Occurrence of class integrons Hospital (N=50) Retirement home (N=50) WWTP influent (N=49) WWTP effuent (N=48) River (N=50)

11 11  In hospital: higher percentages of Enterococcus resistants to high level of erythromycin (70 – 90%), mainly harboring the gene erm(B);  Decrease of hospital-adapted clonal complex CC17 isolates, containing erm(B) and mef(A) genes( p-value<0,001 vs hospital and retirement home) Leclercq et al, 2013 ermB Enterococci resistant to macrolids ( erythromycin) mefA ermB + mefAUnknown genes * * * Changes in Enterococcal populations, and related antibiotic resistance Along the medical center-wastewater treatment plant- river continuum (4km)

12 12 Antibiotic contamination in water along the rural continuum Forest land (Sebec) 50m from a cattle breeding land (450 cattle, Selles) Urban zone (Tourville) Urban zone (Risle) Source: Inhabitants: 396 Cattle : 4592 Anthropic pressure Inhabitants:9058 Catlle: 45

13 13 7, CFU. 100mL -1 Source: 1, CFU. 100mL -1 1, CFU. 100mL -1 1, CFU. 100mL -1 Antibiotic resistance and structure of E.coli population along the rural continuum Anthropic pressure occurence of Shigatoxin producing E.coli Distribution of phylogroups % Antibioresistant E.coli Distribution of phylogroups % Antibioresistant E.coli

14 14 Anthropic pressure Changes in Enterococcal populations, and related antibiotic resistance, along a rural continuum  Increase of abundance of E.faecalis and antibiotic–resistant strains, containing tetM gene from human origins, from downstream to upstream  Environmental species are specifically isolated at the upstream (E. casseliflavus, E.avium, E. gallinarium)

15 15  The contamination of water by antibiotics is related, not only to the medical prescription, but also to the persistence of the molecule of the antibiotic in water environment.  The most persistent antibiotic are fluoroquinolones, macrolides, sulfamides  The concentrations (ng.L -1 ) observed are not sufficient to exert a selective pressure on microbial community (mg.L -1 ) Conclusions Environmental impact of antibiotic use and antibiotic-resistant bacteria Resilience of the estuarine environment  There is a lower survival of culturable antibiotic-resistant E. coli and Enterococcus strains from human (hospital) origin Recommendation ( ANSES september 2012) : With the same therapeutic efficiency, antibiotics which are highly degradated in waters should be used.

16 16 French-Japanese conference of oceanography Boulogne sur Mer 2013 Ph D students Kenny Oberlé UMR M2C/ LPTC, Marion Justine LPTC, Mehdy Ratajczak ( UMR M2C) Laroche Emilie ( UMR M2C) Touron aurélie ( UMR M2C) EC2C0  Hydrology and environmental microbiology UMR CNRS 6143, (M2C), Rouen Thierry Berthe, Nicolas Massei, julien Deloffre, Robert Lafite, Fabienne Petit  Microbiology and epidemiology INSERM U722 Université Paris 7, Faculté de médecine site Xavier Bichat Erick Denamur, Olivier Clermont  and CHU CAEN EA 2125 : Roland Leclercq, Vincent Cattoir  Environmental chemistry UMR CNRS 5255 LPTC- Université Bordeaux Hélène Budzinski


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