Molecular detection of antimicrobial resistance in livestock mycoplasmas: current status and future prospects

Kinga M Sulyok¹Zsuzsa Kreizinger¹Dorottya Földi¹Áron Botond Kovács¹Dénes Grózner¹Lucia Manso-Silvan²Jade Bokma³Annet E. Heuvelink⁴Sara M Klose⁵Anneke Feberwee⁶Salvatore Catania⁷Ana S. Ramirez Corbera⁸Paola K Vaz⁵Cecile Boland⁹Kannan Ganapathy¹⁰Anne V Gautier-Bouchardon¹¹Claire A.M. Becker¹²Florence TARDY¹¹Inna Lysnyansky¹³Miklós Gyuranecz^1*

• ¹HUN-REN Allatorvostudomanyi Kutatointezet, Budapest, Hungary
• ²CIRAD INRAE, UMR ASTRE, Montpellier, France
• ³Universiteit Gent, Ghent, Belgium
• ⁴GD Animal Health, Deventer, Netherlands
• ⁵Asia-Pacific Centre for Animal Health, Faculty of Science, University of Melbourne, Melbourne, Australia
• ⁶Royal GD, Deventer, Netherlands
• ⁷Mycoplasma Unit, WOAH reference laboratory for avian mycoplasmosis, Istituto Zooprofilattico Sperimentale delle Venezie, Buttapietra, Italy
• ⁸IUSA, Veterinary Faculty, University of Las Palmas de Gran Canaria, Gran Canaria, Spain
• ⁹Veterinary Bacteriology, Department of Infectious Diseases in Animals, Sciensano, Brussels, Belgium
• ¹⁰University of Liverpool, Leahurst Campus, Neston, United Kingdom
• ¹¹Anses Laboratoire de Ploufragan-Plouzane, Ploufragan, France
• ¹²UMR Anses VetAgro Sup Mycoplasmoses Animales, VetAgro Sup, Université de Lyon, Marcy l’Etoile, France
• ¹³Kimron Veterinary Institute, Beit Dagan, Israel

Pathogenic Mycoplasma species significantly impact livestock health, causing respiratory, articular, mammary gland, and reproductive disorders with substantial economic losses. Antimicrobials remain essential for controlling clinical signs and production losses; however, treatment efficacy is increasingly threatened by antimicrobial resistance (AMR). Phenotypic methods remain the most reliable approach for detecting AMR in Mycoplasma species; however, they are time-consuming, technically demanding, and results are often difficult to interpret. The absence of clinical breakpoints and limited epidemiological cut-off values (ECOFFs) further complicate AMR categorization. Advances in molecular techniques offer a promising alternative for faster AMR detection and prediction. This review summarizes current knowledge of genetic mechanisms underlying AMR in clinically important Mycoplasma species affecting ruminants, swine, and poultry. It highlights the role of molecular assays in identifying resistance-associated mutations. Additionally, a SWOT (Strengths, Weaknesses, Opportunities, Threats) analysis evaluates these methods' practical applications and limitations in veterinary mycoplasmas. Finally, the potential of genome-wide association studies (GWAS) is explored as an emerging tool for linking genetic traits to phenotypic resistance patterns, offering new insights for enhancing resistance prediction in veterinary medicine.