Epidemiological cut-off values for Vibrio anguillarum MIC and disc diffusion data generated by standardised methods

Peter Smith; Laëtitia Le Devendec; Eric Jouy; Emeline Larvor; Alain Le Breton; Sara Picon-Camacho; Snjeana Zrnčić; Ivana Giovanna Zupičić; Draen Oraić; Süheyla Karataş; David Verner-Jeffreys; Andrew Wokorac Joseph; Edel Light; Alieda van Essen-Zandbergen; Betty van Gelderen; Michal Voorbergen-Laarman; Olga L. M. Haenen; Kees T. Veldman; Lone Madsen; Kári K. Mouritsen; Cecilie Smith Svanevik; Fredrik Håkonsholm; Ana Isabel Vela; María García; Daniela Florio; Marialetizia Fioravanti; Luana Cortinovis; Tobia Pretto; Amedeo Manfrin; Sandrine Baron

doi:10.3354/dao03745

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Diseases of Aquatic Organisms

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DAO 155:109-123 (2023) - DOI: https://doi.org/10.3354/dao03745

DAO Special: 'Epidemiological cut off values for aquatic bacteria'

Epidemiological cut-off values for Vibrio anguillarum MIC and disc diffusion data generated by standardised methods

Peter Smith^1,*, Laëtitia Le Devendec², Eric Jouy², Emeline Larvor², Alain Le Breton³, Sara Picon-Camacho³, Snježana Zrnčić⁴, Ivana Giovanna Zupičić⁵, Dražen Oraić⁴, Süheyla Karataş^5,, David Verner-Jeffreys^6,7, Andrew Wokorac Joseph^6,7, Edel Light⁸, Alieda van Essen-Zandbergen⁹, Betty van Gelderen⁹, Michal Voorbergen-Laarman⁹, Olga L. M. Haenen⁹, Kees T. Veldman⁹, Lone Madsen¹⁰, Kári K. Mouritsen¹⁰, Cecilie Smith Svanevik¹¹, Fredrik Håkonsholm¹¹, Ana Isabel Vela^12,13, María García¹², Daniela Florio¹⁴, Marialetizia Fioravanti¹⁴, Luana Cortinovis¹⁵, Tobia Pretto¹⁵, Amedeo Manfrin¹⁵, Sandrine Baron²

¹School of Natural Science, University of Galway, Galway H91 TK33, Ireland
²Anses, Ploufragan-Plouzané-Niort Laboratory, Mycoplasmology-Bacteriology-Antibioresistance Unit, 22440 Ploufragan, France
³Vet’eau- Selarl du Dr Alain Le Breton, 1989 Rue des Pyrénées, 31330 Grenade sur Garonne, France
⁴Laboratory for Fish and Molluscs Diseases, Croatian Veterinary Institute, 10000 Zagreb, Croatia
⁵Istanbul University, Faculty of Aquatic Sciences, 34234 Istanbul, Turkey
⁶Centre for Environment, Fisheries and Aquaculture Science, Weymouth DT4 8UB, UK
⁷UK FAO Reference Centre for AMR, Weymouth DT4 8UB, UK
⁸Veterinary Medicines Directorate (VMD), Woodham Lane, New Haw, Surrey KT15 3LS, UK
⁹Wageningen Bioveterinary Research, 8221 RA Lelystad, The Netherlands
¹⁰Section for Fish and Shellfish Diseases, National Institute of Aquatic Resources (DTU Aqua), Technical University of Denmark, 2800 Kongens Lyngby, Denmark
¹¹Department of Contaminants and Biohazards, Institute of Marine Research, 5005 Bergen, Norway
¹²Centrode Vigilancia Sanitaria Veterinaria (VISAVET), Universidad Complutense, 28040 Madrid, Spain
¹³Department of Animal Health, Veterinary School, Complutense University, 28040 Madrid, Spain
¹⁴Department of Veterinary Medical Sciences, Alma Mater Studiorum Università di Bologna, 40064, Ozzano Emilia (BO), Italy
¹⁵Fish Pathology Unit, Istituto Zooprofilattico Sperimentale delle Venezie. 35020, Legnaro, Italy

*Corresponding author: peter.smith@universityofgalway.ie

ABSTRACT: This work aims to generate the data needed to set epidemiological cut-off values for minimum inhibitory concentration (MIC) and disc-diffusion zone measurements of Vibrio anguillarum. A total of 261 unique isolates were tested, applying standard methods specifying incubation at 28°C for 24-28 h. Aggregated MIC distributions for a total of 247 isolates were determined in 9 laboratories for 11 agents. Data aggregations of the disc zone for the 10 agents analysed contained between 157 and 218 observations made by 4 to 7 laboratories. Acceptable ranges for quality control (QC) reference strains were available for 7 agents and the related multi-laboratory aggregated data were censored, excluding the data of a laboratory that failed to meet QC requirements. Statistical methods were applied to calculate epidemiological cut-off values. Cut-off values for MIC data were calculated for florfenicol (≤1 µg ml^-1), gentamicin (≤4 µg ml^-1), oxytetracycline (≤0.25 µg ml^-1) and trimethoprim/sulfamethoxazole (≤0.125/2.38 µg ml^-1). The cut-off values for disc zone data were calculated for enrofloxacin (≥29 mm), florfenicol (≥27 mm), gentamicin (≥19 mm), oxolinic acid (≥24 mm), oxytetracycline (≥24 mm) and trimethoprim/sulfamethoxazole (≥26 mm). MIC and disc-diffusion zone data for the other agents where not supported by QC, thus yielding only provisional cut-off values (meropenem, ceftazidime). Regardless of whether QC is available, some of the aggregated MIC distributions (enrofloxacin, oxolinic acid), disc zone (sulfamethoxazole), and MIC and disc-diffusion distributions (ampicillin, chloramphenicol) did not meet the statistical requirements. The data produced will be submitted to the Clinical Laboratory Standards Institute for their consideration in setting international consensus epidemiological cut-off values.

KEY WORDS: CLSI VET04 · Normalized resistance interpretation · ECOFFinder · Antimicrobial susceptibility · Aquatic animals

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Cite this article as: Smith P, Le Devendec L, Jouy E, Larvor E and others (2023) Epidemiological cut-off values for Vibrio anguillarum MIC and disc diffusion data generated by standardised methods. Dis Aquat Org 155:109-123. https://doi.org/10.3354/dao03745

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