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Published on: February 23, 2014
Encapsulation of Disease-Causing and Commensal Mitis Group Non-Pneumococcal Streptococci
Daniel M Musher1,2, Mathias Müsken3, M John Hicks2,4
1Michael E. DeBakey Veterans Affairs Medical Center, Houston, TX 77030, USA.
Abstract:
Background: Mitis group non-pneumococcal streptococci (MGNPS), specifically Streptococcus mitis, Streptococcus infantis and Streptococcus oralis, have recently been shown to cause pneumonia and/or bacteremia. These organisms often have capsular (cps) operons resembling those in pneumococci, and some express cps-generated polysaccharides that antigenically cross-react with pneumococcal capsular serotypes. But, to date, a series of MGNPS isolates has not been studied by electron microscopy (EM) for the presence of a capsule. Methods: We studied 21 MGNPS; 11 were isolated from sputum and determined to have caused pneumonia, 3 were isolated from blood, and 7 were commensal isolates cultured from the oral cavity of healthy adults. Two reacted with a pneumococcal anticapsular antibody. Isolates were fixed with two different protocols and examined by transmission EM. Results: EM of MGNPS after standard fixation and staining with uranyl acetate did not show capsules. In contrast, the 21 MGNPS isolates that we studied after fixation with ruthenium red and lysine acetate were all shown to be encapsulated. The thickness and density of capsules was related to their species: Streptococcus pneumoniae had the most prominent encapsulation and Streptococcus oralis had the least. However, within a species, there was no apparent difference in capsules between disease-causing and colonizing strains. Conclusions: EM with ruthenium red staining demonstrated capsules on 21 MGNPS, but within a species, there was no apparent difference between disease-causing and commensal isolates. It seems reasonable to conclude that the capsule, together with inoculum size, host's ability to clear aspirated organisms, and other as yet unidentified virulence factors, all contribute to the pathogenesis of MGNPS pneumonia.
Insights
Mitis group non-pneumococcal streptococci (MGNPS) possess capsules, revealed by ruthenium red staining. This finding was consistent across disease-causing and commensal strains within species, suggesting capsules contribute to MGNPS pathogenesis.
Area of Science:
- Microbiology
- Bacteriology
- Electron Microscopy
Background:
- Mitis group non-pneumococcal streptococci (MGNPS) are increasingly recognized as causes of pneumonia and bacteremia.
- These bacteria share similarities with Streptococcus pneumoniae, including capsular operons and cross-reactive polysaccharides.
- Previous studies have not utilized electron microscopy to investigate MGNPS encapsulation.
Purpose of the Study:
- To investigate the presence and characteristics of capsules in Mitis group non-pneumococcal streptococci (MGNPS) isolates.
- To compare encapsulation between disease-causing and commensal MGNPS strains.
- To assess the utility of specific electron microscopy fixation techniques for visualizing bacterial capsules.
Main Methods:
- Studied 21 MGNPS isolates (pneumonia, bacteremia, and commensal).
- Employed two fixation protocols for transmission electron microscopy (TEM).
- Utilized ruthenium red and lysine acetate fixation for enhanced capsule visualization.
Main Results:
- Standard TEM fixation failed to reveal capsules on MGNPS.
- Ruthenium red and lysine acetate fixation demonstrated capsules on all 21 MGNPS isolates.
- Capsule thickness and density varied by species, with Streptococcus pneumoniae showing the most prominent encapsulation.
- No significant differences in capsule characteristics were observed between disease-causing and commensal strains within the same species.
Conclusions:
- Ruthenium red staining effectively visualizes capsules on MGNPS, including disease-causing and commensal strains.
- Bacterial capsules are a conserved feature across MGNPS species, though with inter-species variation.
- Capsule presence, along with inoculum size and host factors, likely contributes to the pathogenesis of MGNPS-associated pneumonia.
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