Growth, Congo Red Agar Colony Morphotypes and Antibiotic Susceptibility Testing of Mycobacterium avium subspecies paratuberculosis

• Published in: Clinical medicine & research Vol. 2; no. 2; pp. 107 - 114
• Main Authors: Parrish, Nicole M, Ko, Chiew G, Dick, James D, Jones, Paul B, Ellingson, Jay L E
• Format: Journal Article
• Language: English
• Published: United States Marshfield Clinic 01-05-2004; Marshfield Clinic Research Foundation
• Subjects: Agar; Animals; Cattle; Coloring Agents; Congo Red; Humans; Indicator Dilution Techniques; Microbial Sensitivity Tests - methods; Microbiological Techniques; Mycobacterium avium subsp. paratuberculosis - drug effects; Mycobacterium avium subsp. paratuberculosis - growth & development; Original Research; Paratuberculosis - microbiology
• ISSN: 1539-4182; 1554-6179
• DOI: 10.3121/cmr.2.2.107

Mycobacterium avium subspecies (subsp.) paratuberculosis (MAP) is the causative agent of Johne's disease in ruminants and has been associated with Crohn's disease in humans. We sought to test growth rates and susceptibilities of various strains of MAP in two available growth media.
Paired comparison design.
Using the BACTEC macrobroth radiometric growth system and Congo Red-staining agar media, we determined inherent differences in growth characteristics of three bovine and two human strains of MAP and compared susceptibility results obtained in each growth system.
Significant differences were observed in growth rate as well as mycobactin J dependence between strains and between a laboratory-adapted isolate of the same strain in the macrobroth system. Similarly, colonial morphology and Congo Red staining on agar media were observed. Two strains, one human and one bovine, demonstrated a 100% rough transparent colony with white coloration on Congo Red agar, while one bovine isolate exclusively grew as a smooth opaque colony with red coloration on Congo Red agar. The remaining strains exhibited mixtures of these two colonial morphotypes on agar media. Comparative susceptibility results between the BACTEC radiometric macrobroth method and the agar proportionality method showed good correlation for most antibiotics/inhibitors tested. However, erratic or poor growth in the macrobroth system prevented minimal inhibitory concentration determinations for two bovine strains by this method.
This study demonstrates the variability in the colonial morphology of MAP on Congo Red agar as well as the correlation of antibiotic susceptibility results between the BACTEC macro broth method and the agar proportionality method. This study also emphasizes the need for the development of improved, standardized culture and susceptibility test methods for MAP.