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 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
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 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 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 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 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 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 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 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 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 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 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 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 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 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