SSM_2017 Basel 30/08/2017 Spread of antibiotic resistance markers in Lake Ceresio and tributary rivers. Monitoring results 2016

Federica Mauri Introduction and aims

Monitoring of Ceresio Observe if anthropic lake and tributary rivers activities could influence for the presence of the bacterial populations antibiotic resistance naturally present in water genes and bacteria environments

To gather more information about the role of aquatic ecosystem in spread of antibiotic resistance

2 Introduction

Sampling points

Rivers sampling points (jan-dec)

Rivers influenced by waste water treatment plant

Lake sampling points (feb, may, aug, nov)

Sampling points near potable water uptake plant

3 Results Total cells count: rivers Plate count 1.0E+07

1.0E+06

1.0E+05 CFU/mL 1.0E+04

1.0E+03

1.0E+02 Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec Magliasina Flow cytometry 1.0E+07 Laveggio 1.0E+06

1.0E+05 Cells/ml 1.0E+04

1.0E+03 Jan Feb Mar Apr May Jun Jul Aug Oct Nov Dec 5 Results Total cells count lake Plate count 1.0E+07 1.0E+06 1.0E+05 Gandria 1.0E+04 CFU/mL 1.0E+03 1.0E+02 1.0E+01 Melide 1.0E+00 Feb May Aug Nov Flow Cytometry Ponte 1.0E+07

1.0E+06

1.0E+05

1.0E+04

1.0E+03 Cells/ml 1.0E+02

1.0E+01

1.0E+00 Feb May Aug Nov 4 Results Antibiotic resistance genes: rivers

Rivers influenced by waste water treatment plant

Rivers NOT influenced by waste water treatment plant

FQLs: fluoroquinolones; Bla: β- lactams; Suls: sulphonamides; TETs:

tetracycline 7 Results Antibiotic resistance genes: lake

Absence of resistance genes

Presence of resistance genes

Sampling points Sample type Antibiotics Integrons ABR genes Feb May Aug Nov Gandria Integrated 0-20 m - Class I intI Melide Integrated 0-20 m - Class I intI FQLs - qnrS Integrated 0-20 m - Class I intI Bla - bla (TEM) Lugano 40 m Suls - sulII - Class I intI Riva San Vitale 33 m - Class I intI

FQLs: fluoroquinolones; Bla: β-lactams; Suls: sulphonamides

6 Results qPCR of ABR genes: rivers Genecopies/mL

16S sul II tetA qnrS bla (TEM) 9 Results qPCR of ABR genes: lake

16S

sulII

qnrS

8 Results Antibiotic resistant species

Bla % TETs % Suls % FQLs % Pseudomonas 18 17 24 13 Acinetobacter 19 24 18 25 % of most resistant Enterobacteriaceae 11 13 8 9 strains in rivers Aeromonas 3 3 5 9 Flavobacteriaceae 0.57 1 0.73 0.24 Comamonas 0.34 0.20 0.18 0.24

Bla % TETs % Suls % FQLs % Pseudomonas 13 9 31 3 Microbacterium 3 1.4 5 17 Stenotrophomonas 10 14 10 22 Brevundimonas 1.8 1.7 4 10 % of most resistant Acinetobacter 8 9 14 10 strains in lake Aeromonas 11 18 2.3 6 Enterobatteriacee 4 2 2 0 Sphingobium 2 0 0 0 Flavobacteriaceae 2 5 0 0

FQLs: fluoroquinolones; Bla: β-lactams; Suls: sulphonamides; TETs: tetracycline 10 Conclusions

ARGs were most sulII, qnrS, tetA present in rivers are the most influenced by abundant genes WWTP and in the lake near the coast .

Rivers and lakes are reservoir of Pseudomonas , Acinetobacter , Anthropic impact resistance genes Aeromonas , and resistance Enterobacteriaceae, bacteria Stenotrophomonas,

intI in all sampling points Mobilization of ARGs

11 Many Thanks to:

Prof. Dr Mauro Tonolla Laboratory of Applied Microbiology SUPSI Dr. Antonella Demarta Laboratory of Applied Microbiology SUPSI Dr. Andreas Bruder Institute of Earth Sciences SUPSI Dr. Giuseppe Ranieri Ufficio monitoraggio ambientale (SPAAS) Dr. Nicola Solcà Ufficio della gestione dei rischi ambientali e del suolo Dr. Gianluca Corno National Research Council of (CNR)

Thank you for your attention! Results Antibiotic resistance genes: rivers

Antibiotics ABR genes jan feb mar apr may jun jul aug sep oct nov dec Antibiotics ABR genes jan feb mar apr may jun jul augsep oct nov dec bla (TEM) bla (TEM) bla (SHV) bla (SHV) β-lactams β-lactams bla ( CTX-M) bla ( CTX-M) bla (oxa 48) bla (oxa 48) qnrA qnrA Fluoroquinolones Fluoroquinolones qnrS qnrS tetA tetA Vedeggio Tetracycline tetM Cassarate Tetracycline tetM tetB tetB sulII sulII Sulphonamides Sulphonamides sulIII sulIII

Integrase intI Integrase intI

Antibiotics ABR genes jan feb mar apr may jun jul aug sep oct nov dec Antibiotics ABR genes jan feb mar apr may jun jul augsep oct nov dec bla (TEM) bla (TEM) bla (SHV) bla (SHV) β-lactams β-lactams bla ( CTX-M) bla ( CTX-M) bla (oxa 48) bla (oxa 48) qnrA qnrA Fluoroquinolones Fluoroquinolones qnrS qnrS tetA tetA Laveggio Tetracycline tetM Tetracycline tetM Magliasina tetB tetB sulII sulII Sulphonamides Sulphonamides sulIII sulIII

Integrase intI Integrase intI

Absence of resistance genes

Presence of resistance genes