Insights from 17 years of culture and PCR detection of animal mollicutes in a Canadian provincial laboratory.

Since ~1980, the Animal Health Laboratory (AHL) in Ontario, Canada, has isolated animal mollicute species by culture. Data for the most recent 17 y (2007-2024) captures over 90,000 test results. Advancements in PCR, qPCR, and DNA sequencing have shifted the percentage of testing by PCR from 18.7% in 2007 to 91.1% in 2024. The bulk of this shift is due to the uptake of molecular testing as a screening tool for clinically normal animals, but this shift has not been universal, particularly for ureaplasma testing. Culture remains the gold standard for the detection and identification of rare pathogens and plays a key role in research through our mycoplasma cryobank, which includes 40+ y of isolates. Synergizing the microbiologic and molecular techniques developed over the AHL's multi-decade history has presented novel opportunities for detection, characterization, and local eradication of animal mollicutes, including the development of new assays, tracking of historical trends for antimicrobial resistance (AMR), and identifying AMR-associated mutations in Mycoplasmopsis (Mycoplasma) bovis.