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Growth, Congo Red agar colony morphotypes and antibiotic susceptibility testing of Mycobacterium avium subspecies
Nicole M Parrish1, Chiew G Ko, James D Dick
1Department of Pathology, School of Medicine, Johns Hopkins University, Baltimore, Maryland, USA.
Objective:
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.
Design:
Paired comparison design.
Methods:
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.
Results:
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.
Conclusion:
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.
Insights
Mycobacterium avium subspecies (MAP) strains show varied growth and colony types on different media. Antibiotic susceptibility testing correlated between methods, but improved diagnostic approaches for MAP are needed.
Area of Science:
- Veterinary Microbiology
- Human Infectious Diseases
- Bacteriology
Background:
- Mycobacterium avium subspecies (subsp.) paratuberculosis (MAP) causes Johne's disease in ruminants.
- MAP has been linked to Crohn's disease in humans.
- Variability in MAP growth and susceptibility testing poses diagnostic challenges.
Purpose of the Study:
- To compare growth rates and susceptibilities of different MAP strains.
- To evaluate two distinct culture media for MAP analysis.
- To assess the correlation of antibiotic susceptibility results between two testing systems.
Main Methods:
- Utilized the BACTEC macrobroth radiometric growth system and Congo Red-staining agar media.
- Compared growth characteristics of three bovine and two human MAP strains.
- Assessed colonial morphology and mycobactin J dependence on agar and in broth.
Main Results:
- Observed significant differences in MAP growth rates and mycobactin J dependence between strains.
- Identified distinct colonial morphotypes (rough transparent vs. smooth opaque) on Congo Red agar.
- Demonstrated good correlation in antibiotic susceptibility results between the BACTEC system and agar proportionality method, with some limitations.
Conclusions:
- MAP exhibits significant variability in colonial morphology on Congo Red agar.
- The BACTEC macrobroth and agar proportionality methods show correlated antibiotic susceptibility results.
- There is a critical need for enhanced and standardized diagnostic methods for MAP.
