Discrimination between Mycoplasma and Acholeplasma species of bovine origin using digitonin disc diffusion assay, nisin disc diffusion assay, and conventional polymerase chain reaction

Sukolrat Boonyayatra; Lawrence K Fox; John M Gay; Ashish Sawant; Thomas E Besser

doi:10.1177/1040638711425936

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Discrimination between Mycoplasma and Acholeplasma species of bovine origin using digitonin disc diffusion assay, nisin disc diffusion assay, and conventional polymerase chain reaction

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Discrimination between Mycoplasma and Acholeplasma species of bovine origin using digitonin disc diffusion assay, nisin disc diffusion assay, and conventional polymerase chain reaction

Sukolrat Boonyayatra, Lawrence K Fox, John M Gay, Ashish Sawant and Thomas E Besser

Journal of veterinary diagnostic investigation, Vol.24(1), pp.7-13

01/2012

DOI: https://doi.org/10.1177/1040638711425936

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https://hdl.handle.net/2376/112822

PMID: 22362930

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Abstract

Acholeplasma laidlawii - genetics

Disk Diffusion Antimicrobial Tests - methods

Nisin

Cattle Diseases - microbiology

Mycoplasma Infections - microbiology

Mycoplasma - genetics

Mycoplasma Infections - diagnosis

Disk Diffusion Antimicrobial Tests - veterinary

Polymerase Chain Reaction - methods

Gram-Negative Bacterial Infections - microbiology

Animals

Polymerase Chain Reaction - veterinary

Acholeplasma - genetics

Cattle

Mycoplasma Infections - veterinary

RNA, Ribosomal, 16S - genetics

Female

Gram-Negative Bacterial Infections - diagnosis

Cattle Diseases - diagnosis

Gram-Negative Bacterial Infections - veterinary

Digitonin

Microbiological culture of milk samples has been used as a standard diagnosis for Mycoplasma mastitis. This technique is effective in isolating mollicutes that are Mycoplasma-like; however, isolates may be misinterpreted as Acholeplasma species, which are indistinguishable from Mycoplasma species by culture. A study to contrast the abilities of 2 culture-based tests, digitonin and nisin disc diffusion assays and a conventional polymerase chain reaction (PCR) technique, to discriminate between Mycoplasma and Acholeplasma was performed using 16S ribosomal RNA gene partial sequencing as the gold standard of comparison. A total of 288 bovine mollicute field isolates (248 from milk and 40 from other organ sites) and 13 reference strains were tested. Results obtained from the digitonin disc diffusion assay when it was performed with all field isolates were 92.7% and 99.0% in agreement with the gold standard using 5 mm and 3 mm of zone of growth inhibition as thresholds, respectively. Considering only milk isolates, agreements between the digitonin disc diffusion assay with the gold standard were 97.2% and 100% using 5 mm and 3 mm of zone of growth inhibition as thresholds, respectively. Culture identification using the nisin disc diffusion assay and the PCR was in a 100% agreement with the gold standard. Comparable results using culture-based nisin and digitonin disc diffusion assays, and PCR, to distinguish Mycoplasma and Acholeplasma species was found, especially for isolates from bovine milk.

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Title: Discrimination between Mycoplasma and Acholeplasma species of bovine origin using digitonin disc diffusion assay, nisin disc diffusion assay, and conventional polymerase chain reaction
Creators: Sukolrat Boonyayatra - Department of Veterinary Clinical Science, College of Veterinary Medicine, Washington State University, Pullman, WA 99164, USA
Lawrence K Fox
John M Gay
Ashish Sawant
Thomas E Besser
Publication Details: Journal of veterinary diagnostic investigation, Vol.24(1), pp.7-13
Academic Unit: Veterinary Clinical Sciences, Department of; Veterinary Microbiology and Pathology, Department of
Publisher: United States
Identifiers: 99900547623001842
Language: English
Resource Type: Journal article