An outbreak of STEC O26 linked to water
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1 12 th Annual Workshop of the NRLs for E. coli in the EU ottobre 2017 An outbreak of STEC O26 linked to water Analysis on food, animal faeces and water Conedera G., Pierasco A., Targhetta C., Vio D. IZS Venezie, Laboratory of Pordenone Ceschi P. Veterinary Service, Local Health Unit, Bolzano
2 Aim of the presentation To describe the laboratory analysis to investigate STEC O26 in foods, animal faeces and water, sampled in the area of a small municipality of Bolzano province, within the epidemiological survey on HUS cases occurred between June and July 2017
3 Methods Food samples ISO/TS 13136:2012 (25gr/ ml) SAMPLING performed by Veterinary Service and Hygiene PH Service of the Local Health Unit of Bolzano Water samples One liter of water, filtered through membranes (0.45 µm) Membranes transferred in 100 ml BPW (18-24h at 37ºC) Nucleic acid extraction and detection according to ISO/TS 13136:2012 Fecal samples 10 gr of faeces, pre-enrichment at 37 º C in 90 ml BPW for 18-24h Nucleic acid extraction from 1 ml of microbial enrichment with Instagene Matrix (Biorad) Nucleic acid diluted 1:20 prior to real time amplification Detection of serogroup O26 gene according to ISO/TS 13136:2012 For O26 positive samples, detection of stx1/stx2/eae genes according to ISO/TS 13136:2012 For STEC O26 positive samples, isolation of microrganism according to ISO/TS 13136:2012
4 Sampling at MOUNTAIN PASTURE FARM with small dairy factory and B&B (1930 m asl) FOOD (July) 1 bulk milk sample 7 dairy products NO STEC isolation FECAL SAMPLES Sampling at MP farm (August) 24 individual fecal samples (animals from 3 different farms) Sampling at original farms (September) (after the animals came back from the summer pastures) 39 fecal samples from herd 1 6 fecal samples from herd 2 9 fecal samples from herd 3 Isolation of E. coli O26 stx2 eae from 1/24 cows (herd 1) Isolation of E. coli O26 stx2 eae from 5/39 animals (2 calves and 3 heifers) of herd 1 only Presence of O26 and stx genes in other 7 animals of herd 1 only
5 Sampling at ALPINE DAIRY FARM with B&B (1300m asl) FOOD (July) 1 bulk milk sample 3 ice cream samples (not homemade) (5 more ice creams sampled by the same local producer) NO STEC isolation FECAL SAMPLES (July) 45 individual bovine fecal samples 2 individual goat fecal samples NO STEC isolation
6 Further sampling (July - September) 3 SMALL TRADITIONAL DAIRY FARMS (July- September): a total of 31 bovine fecal samples and 3 bulk milk samples 6 bovine cow pats collected in the close proximity of a small stream (September) WILD LIFE (September): 3 red deer fecal samples NO STEC isolation
7 Water samples (end of July) 11 WATER SAMPLES FROM MOUNTAIN SPRINGS, TANKS, FOUNTAINS AND AQUADUCTS RESULTS STEC were isolated from 4/11 water samples: E. coli O26 stx2 eae from 2 water samples from the same mountain area with forest (one from a public aquaduct and one from a private spring close to alpine dairy farm with B&B), apparently not connected E. coli O26 stx1 eae from 1 water sample collected from a fountain near a small dairy farm STEC stx1 stx2 from 1 water sample from a spring near the mountain pasture farm with B&B
8 PFGE at EU-RL highlighted 100% omology between the profile of E. coli O26 stx2 eae strains isolated from the 2 water samples and the human strains (isolated by AGES-Austria and ISS and SouthTirol Labs) apart from 2 (one of which anyway was very similar) by Antonella Maugliani Bovine strain O26 stx2 eae from mountain pasture farm with B&B was not related with HUS strains
9 In summary we tested: 156 bovine fecal samples 2 goat fecal samples 3 red deer fecal samples 11 water samples 20 food samples Conclusions STEC were isolated from 4/11 water samples 2 STEC O26 isolated from water with 100% omology with human STEC O26 HUS strains Bovine strain STEC O26 from mountain pasture farm with B&B was not related with STEC O26 HUS strains Cluster of STEC O26 cases apparently linked to water Further investigations are required
10 Discussion: possible points raised. Analytical methods for STEC isolation from environment Mountain water springs contamination and risk of infection for local population and tourists, considering in particular young children Management of water resources and contamination prevention Possibile source of STEC O26 strains domestic/ wild ruminants, (humans)? Role of cattle as a reservoir of STEC O26 and STEC O26 ecology within the farm Need of prompt pathogen detection and source identification Importance of communication and cooperation among LNRs and National Authorities of different countries (good communication between LNR Austria and Italy) Need of continuous exchange of information and cooperation among Local and National Health Authorities involved in clinical diagnostics, microbiological and epidemiological surveys
11 Aknowledgements IZS Venezie staff Pordenone EURL E. coli, ISS Rome IZS Venezie - Bolzano Local Health Unit Bolzano Veterinary Service Dr. A Fugatti Hygiene and Public Health Service Dott.ssa MG Zuccaro and their staff Thank you!
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