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Home / Equine Research Bank / Equine Vet J / Detection of Mycoplasma spp. in horses with respiratory diso...
Equine veterinary journal2023; 55(5); 747-754; doi: 10.1111/evj.13918

Detection of Mycoplasma spp. in horses with respiratory disorders.

Abstract: Bacteria belonging to the genus Mycoplasma are small-sized, have no cell walls and small genomes. They commonly cause respiratory disorders in their animal hosts. Three species have been found in the respiratory tract of horses worldwide, that is., Mycoplasma (M.) equirhinis, M. pulmonis and M. felis, but their role in clinical cases remains unclear. Objective: The aim of this study was to i) develop and validate tools to detect, isolate and identify different Mycoplasma spp. strains in clinical equine respiratory-tract specimens and ii) subsequently define the prevalence of the three species in France depending on sample types and horse characteristics (age, breed, sex). Methods: Validation of a workflow for mycoplasma diagnosis and subsequent prevalence study. Methods: Mycoplasma-free tracheal wash samples spiked with numerated strains and DNA dilutions were used to validate the culture methods and real-time PCR (rt-PCR) assay. Isolated strains were identified by 16S rRNA gene sequencing. Prevalences were determined on a population of 616 horses with respiratory disorders, sampled in France in 2020. Results: In total, 104 horses (16.9%) were found to be positive for Mycoplasma spp. by at least one method. M. equirhinis was the predominant circulating species, accounting for 85% of the rt-PCR-positive samples and 98% of the 40 cultured strains. Conclusions: The proposed pre-enrichment procedure improves the sensitivity of detection but hinders the quantification of the initial mycoplasma load in the clinical specimens. Conclusions: Prevalence of mycoplasma varied with age, breed, and type of sample. Unassigned: Bakterien, die zur Gattung Mycoplasma gehören, sind klein, haben keine Zellwände und ein kleines Genom. Sie verursachen bei ihren tierischen Wirten häufig Erkrankungen der Atemwege. Drei Spezies wurden weltweit in den Atemwegen von Pferden gefunden, nämlich Mycoplasma (M.) equirhinis, M. pulmonis und M. felis, aber ihre Rolle in klinischen Fällen bleibt unklar. Unassigned: Ziel dieser Studie war es, i) Instrumente zum Nachweis, zur Isolierung und zur Identifizierung verschiedener Mycoplasma spp. in klinischen Atemwegsproben von Pferden zu entwickeln und zu validieren und ii) anschließend die Prävalenz der drei Spezies in Frankreich in Abhängigkeit von Probentypen und Pferdemerkmalen (Alter, Rasse, Geschlecht) zu bestimmen. Methods: Validierung eines Arbeitsablaufs für die Mykoplasmen-Diagnose und anschließende Prävalenzstudie. Methods: Mykoplasmenfreie Trachealspülungen, die mit nummerierten Stämmen und DNA-Verdünnungen versetzt waren, wurden zur Validierung der Kulturmethoden und des real-time PCR-Tests (rt-PCR) verwendet. Isolierte Stämme wurden durch Sequenzierung des 16S rRNA-Gens identifiziert. Die Prävalenzen wurden in einer Population von 616 Pferden mit Atemwegserkrankungen bestimmt, die 2020 in Frankreich beprobt wurden. Unassigned: Insgesamt wurden 104 Pferde (16.9%) mit mindestens einer Methode positiv auf Mycoplasma spp. getestet. M. equirhinis war die vorherrschende zirkulierende Spezies und machte 85% der rt-PCR-positiven Proben und 98% der 40 kultivierten Stämme aus. Unassigned: Das vorgeschlagene Voranreicherungsverfahren verbessert die Nachweisempfindlichkeit, erschwert aber die Quantifizierung der ursprünglichen Mykoplasmenbelastung in den klinischen Proben. Unassigned: Die Prävalenz von Mykoplasmen variierte je nach Alter, Status des Rennpferdes und Art der Probe.
© 2022 The Authors. Equine Veterinary Journal published by John Wiley & Sons Ltd on behalf of EVJ Ltd.
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Publication Date: 2023-01-09 PubMed ID: 36572918DOI: 10.1111/evj.13918Google Scholar: Lookup
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Summary

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The research article investigates the detection and prevalence of the Mycoplasma species, a bacteria that typically causes respiratory disorders in horses. By developing a process to discover and isolate the bacteria, and then applying this to a study sample of horses in France, the research provides a deeper understanding of the bacteria’s presence and impact.

Objective and Approach

  • The study’s main objective was to develop a reliable method to detect, isolate, and identify different strains of the Mycoplasma bacteria from respiratory-tract specimens of horses, and to determine the prevalence of the three main species across varying horse characteristics (like age, breed, sex) in France.
  • The researchers validated a diagnostic workflow for mycoplasma bacteria and then used this to conduct a prevalence study.

Methodology

  • Tracheal wash samples free of Mycoplasma were spiked with strains and DNA dilutions, allowing the researchers to validate their culture methods and real-time PCR (rt-PCR) assay – a laboratory technique used to amplify and simultaneously quantify a targeted DNA molecule.
  • The scientists identified individual strains by sequencing the 16S rRNA gene, a component within the small subunit of ribosomes in the bacteria.
  • The prevalence investigation involved a population of 616 horses with respiratory disorders in France sampled in 2020.

Findings and Conclusions

  • Of the horse sample, 104 horses (16.9%) tested positive for Mycoplasma species using at least one detection method.
  • The predominant species among the horses sampled was M. equirhinis, representing 85% of the rt-PCR-positive samples and 98% of the 40 cultivated strains.
  • The researchers developed a pre-enrichment procedure that improved detection sensitivity. However, the technique hampers the quantification of the initial Mycoplasma load in the clinical specimens, making it difficult to establish the initial bacterial load when a horse has been infected.
  • The prevalence of Mycoplasma varied with factors such as the age of the horses, breed, and type of sample collected. This suggests that various factors could affect the susceptibility of horses to Mycoplasma infection.

Cite This Article

APA
Martineau M, Castagnet S, Kokabi E, Tricot A, Jaÿ M, Léon A, Tardy F. (2023). Detection of Mycoplasma spp. in horses with respiratory disorders. Equine Vet J, 55(5), 747-754. https://doi.org/10.1111/evj.13918

Publication

Equine veterinary journal
ISSN: 2042-3306
NlmUniqueID: 0173320
Country: United States
Language: English
Volume: 55
Issue: 5
Pages: 747-754

Researcher Affiliations

Martineau, Matthieu
  • LABÉO, Research Department, St Contest, Caen, France.
  • NormandieUniversité, CAEN/ROUEN Universités, DYNAMICURE, INSERM U1311, France.
  • Université de Lyon, Anses, VetAgro Sup, UMR Mycoplasmoses Animales, Lyon, France.
Castagnet, Sophie
  • LABÉO, Research Department, St Contest, Caen, France.
  • NormandieUniversité, CAEN/ROUEN Universités, DYNAMICURE, INSERM U1311, France.
Kokabi, Elena
  • LABÉO, Research Department, St Contest, Caen, France.
  • NormandieUniversité, CAEN/ROUEN Universités, DYNAMICURE, INSERM U1311, France.
Tricot, Agnès
  • Université de Lyon, Anses, VetAgro Sup, UMR Mycoplasmoses Animales, Lyon, France.
Jaÿ, Maryne
  • Université de Lyon, Anses, VetAgro Sup, UMR Mycoplasmoses Animales, Lyon, France.
Léon, Albertine
  • LABÉO, Research Department, St Contest, Caen, France.
  • NormandieUniversité, CAEN/ROUEN Universités, DYNAMICURE, INSERM U1311, France.
Tardy, Florence
  • Université de Lyon, Anses, VetAgro Sup, UMR Mycoplasmoses Animales, Lyon, France.

MeSH Terms

  • Horses / genetics
  • Animals
  • Mycoplasma / genetics
  • Mycoplasma Infections / epidemiology
  • Mycoplasma Infections / veterinary
  • Mycoplasma Infections / microbiology
  • RNA, Ribosomal, 16S / genetics
  • Respiratory Tract Diseases / epidemiology
  • Respiratory Tract Diseases / veterinary
  • Trachea / microbiology

Grant Funding

  • Fonds Eperon
  • Groupement d'Intu00e9ru00eat Scientifique Centaure
  • Institut Franu00e7ais du Cheval et de l'Equitation

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Citations

This article has been cited 3 times.
  1. Hardefeldt L, Thomas K, Page S, Norris J, Browning G, El Hage C, Stewart A, Gilkerson J, Muscatello G, Verwilghen D, van Galen G, Bauquier J, Cuming R, Reynolds B, Whittaker C, Wilkes E, Clulow J, Burden C, Begg L. Antimicrobial prescribing guidelines for horses in Australia. Aust Vet J 2025 Dec;103(12):781-889.
    doi: 10.1111/avj.70003pubmed: 40903020google scholar: lookup
  2. Simms N, Bertone JJ, Melgarejo T, O'Shea C, Linde A. Equine Blood Microbiome in a Cohort of Clinically Healthy Trail Riding Horses. J Vet Intern Med 2025 May-Jun;39(3):e70082.
    doi: 10.1111/jvim.70082pubmed: 40167194google scholar: lookup
  3. Kinoshita Y, Niwa H, Uchida-Fujii E, Ueno T. A real-time PCR assay for the quantification of Mycoplasma equirhinis in tracheal wash samples from Thoroughbred horses. J Vet Diagn Invest 2024 Jan;36(1):108-111.
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