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Diseases of Aquatic Organisms

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DAO 155:109-123 (2023)  -  DOI:

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

Peter Smith1,*, Laëtitia Le Devendec2, Eric Jouy2, Emeline Larvor2, Alain Le Breton3, Sara Picon-Camacho3, Snježana Zrnčić4, Ivana Giovanna Zupičić5, Dražen Oraić4, Süheyla Karataş5,, David Verner-Jeffreys6,7, Andrew Wokorac Joseph6,7, 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 Vela12,13, 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:

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.

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