Mutans streptococci enumeration and genotype selection using different bacitracin-containing media

Stephanie S Momeni¹, Paul Patrick², Howard W Wiener³, Gary R Cutter⁴, John D Ruby⁵, Kyounga Cheon⁶, Jennifer Whiddon⁷, Stephen A Moser⁸, Noel K Childers⁹

Affiliations

¹ Department of Pediatric Dentistry, School of Dentistry, University of Alabama at Birmingham, SOD 304, 1720 2nd Avenue South, Birmingham, AL 35294-0007, USA. Electronic address: sks@uab.edu.
² Department of Pediatric Dentistry, School of Dentistry, University of Alabama at Birmingham, SOD 304, 1720 2nd Avenue South, Birmingham, AL 35294-0007, USA. Electronic address: paul.patrick53@yahoo.com.
³ Department of Epidemiology, School of Public Health, University of Alabama at Birmingham, RPHB 217D, 1720 2nd Avenue South, Birmingham, AL 35294-0022, USA. Electronic address: hwiener@uab.edu.
⁴ Department of Biostatistics, School of Public Health, University of Alabama at Birmingham, RPHB 327, 1720 2nd Avenue South, Birmingham, AL 35294-0022, USA. Electronic address: cutterg@uab.edu.
⁵ Department of Pediatric Dentistry, School of Dentistry, University of Alabama at Birmingham, SOD 304, 1720 2nd Avenue South, Birmingham, AL 35294-0007, USA. Electronic address: johnruby@uab.edu.
⁶ Department of Pediatric Dentistry, School of Dentistry, University of Alabama at Birmingham, SOD 304, 1720 2nd Avenue South, Birmingham, AL 35294-0007, USA. Electronic address: kcheon@uab.edu.
⁷ Department of Pathology, School of Medicine, University of Alabama at Birmingham, WP P230, 1720 2nd Avenue South, Birmingham, AL 35294-7331, USA. Electronic address: jenndoc@uab.edu.
⁸ Department of Pathology, School of Medicine, University of Alabama at Birmingham, WP P230, 1720 2nd Avenue South, Birmingham, AL 35294-7331, USA. Electronic address: moser@uab.edu.
⁹ Department of Pediatric Dentistry, School of Dentistry, University of Alabama at Birmingham, SOD 304, 1720 2nd Avenue South, Birmingham, AL 35294-0007, USA. Electronic address: nkc@uab.edu.

PMID: 24878141
PMCID: PMC4121667
DOI: 10.1016/j.mimet.2014.05.010

Mutans streptococci enumeration and genotype selection using different bacitracin-containing media

Stephanie S Momeni et al. J Microbiol Methods. 2014 Aug.

. 2014 Aug:103:53-7.

doi: 10.1016/j.mimet.2014.05.010. Epub 2014 May 28.

Authors

Stephanie S Momeni¹, Paul Patrick², Howard W Wiener³, Gary R Cutter⁴, John D Ruby⁵, Kyounga Cheon⁶, Jennifer Whiddon⁷, Stephen A Moser⁸, Noel K Childers⁹

Affiliations

¹ Department of Pediatric Dentistry, School of Dentistry, University of Alabama at Birmingham, SOD 304, 1720 2nd Avenue South, Birmingham, AL 35294-0007, USA. Electronic address: sks@uab.edu.
² Department of Pediatric Dentistry, School of Dentistry, University of Alabama at Birmingham, SOD 304, 1720 2nd Avenue South, Birmingham, AL 35294-0007, USA. Electronic address: paul.patrick53@yahoo.com.
³ Department of Epidemiology, School of Public Health, University of Alabama at Birmingham, RPHB 217D, 1720 2nd Avenue South, Birmingham, AL 35294-0022, USA. Electronic address: hwiener@uab.edu.
⁴ Department of Biostatistics, School of Public Health, University of Alabama at Birmingham, RPHB 327, 1720 2nd Avenue South, Birmingham, AL 35294-0022, USA. Electronic address: cutterg@uab.edu.
⁵ Department of Pediatric Dentistry, School of Dentistry, University of Alabama at Birmingham, SOD 304, 1720 2nd Avenue South, Birmingham, AL 35294-0007, USA. Electronic address: johnruby@uab.edu.
⁶ Department of Pediatric Dentistry, School of Dentistry, University of Alabama at Birmingham, SOD 304, 1720 2nd Avenue South, Birmingham, AL 35294-0007, USA. Electronic address: kcheon@uab.edu.
⁷ Department of Pathology, School of Medicine, University of Alabama at Birmingham, WP P230, 1720 2nd Avenue South, Birmingham, AL 35294-7331, USA. Electronic address: jenndoc@uab.edu.
⁸ Department of Pathology, School of Medicine, University of Alabama at Birmingham, WP P230, 1720 2nd Avenue South, Birmingham, AL 35294-7331, USA. Electronic address: moser@uab.edu.
⁹ Department of Pediatric Dentistry, School of Dentistry, University of Alabama at Birmingham, SOD 304, 1720 2nd Avenue South, Birmingham, AL 35294-0007, USA. Electronic address: nkc@uab.edu.

PMID: 24878141
PMCID: PMC4121667
DOI: 10.1016/j.mimet.2014.05.010

Abstract

The primary etiological agents associated with dental caries include the mutans streptococci (MS) comprised of Streptococcus mutans and Streptococcus sobrinus. The effective cultivation and isolation of MS are necessary for the study of MS, including their proper clinical assessment in the epidemiological study of dental caries. Several selective media have been developed for the isolation, enumeration, and characterization of MS. However, inhibition of MS may occur, reducing counts and perhaps limiting selection of some strains. The purpose of this study was to compare five culture media containing bacitracin recommended for the isolation of MS. Five commonly used bacitracin-containing media (MSB, MSKB, GTSB, TYS20B, and TYCSB) used for MS isolation were quantitatively evaluated. Standard plate counts were performed in duplicate for 2 prototype MS strains (S. mutans UA159 and S. sobrinus 6715) and for MS isolates from clinical saliva samples obtained from 16 children (approximate age 5years) to determine total plate counts, and total S. mutans counts. Selected isolates (n=249) from all five media for 5 saliva samples were further confirmed as S. mutans with real-time PCR then subsequently evaluated qualitatively with rep-PCR for genotype determination. All media resulted in variable enumeration with no significant difference in MS counts. MS prototype strains grew well on all five media; clinical isolates demonstrated more variability in counts but no overall significant differences were found. MSB demonstrated comparable ability to grow S. mutans but allowed for more non-S. mutans growth. All 5 media identified a consistent predominant genotype by rep-PCR. Recovery of minor genotypes was not inhibited by media type.

Keywords: Genotypes; Selective media; Streptococcus mutans; Streptococcus sobrinus; Total streptococci; rep-PCR.

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Conflict of interest statement

The authors declare no conflicts of interest.

References

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Mutans streptococci enumeration and genotype selection using different bacitracin-containing media

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Mutans streptococci enumeration and genotype selection using different bacitracin-containing media

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