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. 2002 Feb;68(2):820-30.
doi: 10.1128/AEM.68.2.820-830.2002.

Biodiversity of the bacterial flora on the surface of a smear cheese

Noelle M Brennan  1 , Alan C Ward, Thomas P Beresford, Patrick F Fox, Michael Goodfellow, Timothy M Cogan
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V体育安卓版 - Biodiversity of the bacterial flora on the surface of a smear cheese

VSports最新版本 - Noelle M Brennan et al. Appl Environ Microbiol. 2002 Feb.

"VSports最新版本" Abstract

The bacteria on the surface of a farmhouse smear-ripened cheese at four stages of ripening (4, 16, 23, and 37 days) from inoculated (i. e. , deliberately inoculated with Brevibacterium linens BL2) and noninoculated (not deliberately inoculated with B. linens BL2) cheese were investigated. The results show that, contrary to accepted belief, B. linens is not a significant member of the surface flora of smear cheese and no microbial succession of species occurred during the ripening of the cheeses. Of 400 isolates made, 390 were lactate-utilizing coryneforms and 10 were coagulase-negative Staphylococcus spp. A detailed analysis of the coryneforms was undertaken using phenotypic analysis, molecular fingerprinting, chemotaxonomic techniques, and 16S rRNA gene sequencing. DNA banding profiles (ramdom amplified polymorphic DNA [RAPD]-PCR) of all the coryneform isolates showed large numbers of clusters. However, pulsed-field gel electrophoresis (PFGE) of the isolates from the cheeses showed that all isolates within a cluster and in many contiguous clusters were the same. The inoculated and noninoculated cheeses were dominated by single clones of novel species of Corynebacterium casei (50. 2% of isolates), Corynebacterium mooreparkense (26% of isolates), and Microbacterium gubbeenense (12. 8% of isolates). In addition, five of the isolates from the inoculated cheese were Corynebacterium flavescens. Thirty-seven strains were not identified but many had similar PFGE patterns, indicating that they were the same species. C VSports手机版. mooreparkense and C. casei grew at pH values below 4. 9 in the presence of 8% NaCl, while M. gubbeenense did not grow below pH 5. 8 in the presence of 5 to 10% NaCl. B. linens BL2 was not recovered from the inoculated cheese because it was inhibited by all the Staphylococcus isolates and many of the coryneforms. It was concluded that within a particular batch of cheese there was significant bacterial diversity in the microflora on the surface. .

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Figures

FIG. 1.
FIG. 1.
Development of bacteria and yeast on the surface of the inoculated and noninoculated cheese during ripening.
FIG. 2.
FIG. 2.
(A) Salt and moisture levels in the surface layer of the inoculated and noninoculated cheese during ripening. (B) Salt-in-moisture levels and pH in the surface layers of the inoculated and noninoculated cheese during ripening.
FIG. 3.
FIG. 3.
Reproducibility of the RAPD-PCR profiles of several culture collection strains of coryneform bacteria, most of which have been isolated from cheese. D1, D2, etc. refer to the day on which the PCRs were performed and A and B refer to duplicate PCRs of that strain.
FIG. 4.
FIG. 4.
Comparison of RAPD-PCR results and PFGE band patterns of isolates from the inoculated cheese. The cluster numbers (C1 to C29) are shown and the number of strains in each cluster is in parentheses. All isolates within a cluster had the same PFGE pattern and, for clarity, only one example of the band pattern in each cluster is shown. Strains which were previously identified by chemotaxonomic analyses and 16S rRNA sequencing (1, 2) are also indicated, while strains with one asterisk were identified in this study by chemotaxonomic analyses and 16S rRNA sequencing.
FIG. 5.
FIG. 5.
Comparison of RAPD-PCR results and PFGE band patterns of isolates from the noninoculated cheese. The cluster numbers (C1 to C26) are shown and the number of strains in each cluster is in parentheses. All isolates within a cluster had the same PFGE pattern and, for clarity, only one example of the pattern in each cluster is shown. Strains which were previously identified by chemotaxonomic analyses and 16S rRNA sequencing (1, 2) are also indicated.
FIG. 6.
FIG. 6.
Growth of M. gubbeenense DPC 5283 (A), C. mooreparkense DPC 5308 (B), and C. casei DPC 5583 (C) on 2-dimensional pH and salt gradient plates. The dark areas indicate where growth occurred and the light areas indicate where growth did not occur.
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"V体育2025版" References

    1. Brennan, N. M., R. Brown, M. Goodfellow, A. C. Ward, T. P. Beresford, P. J. Simpson, P. F. Fox, and T. M. Cogan. 2001. Corynebacterium mooreparkense sp. nov., and Corynebacterium casei sp. nov. isolated from the surface of a smear-ripened cheese. Int. J. Syst. E vol. Microbiol. 51:843-852. - PubMed
    1. Brennan, N. M., R. Brown, M. Goodfellow, A. C. Ward, T. P. Beresford, M. Vancanneyt, T. M. Cogan, and P. F. Fox. 2001. Microbacterium gubbeenense sp. nov., isolated from the surface of a smear-ripened cheese. Int. J. Syst. Evol. Microbiol. 51:1969-1976. - PubMed
    1. Busse, M. 1989. Die Oberflachenflora von Geschmierten Käse. Milchwirt. Berichte. 99:137-141.
    1. Cure, G. L., and R. M. Keddie. 1973. Methods for the morphological examination of aerobic coryneform bacteria, p. 123-135. In R. G. Board and D. W. Lovelock (ed.), Sampling—microbiological monitoring of environments—1973. Academic Press, London, United Kingdom.
    1. El-Erian, A. F. M. 1969. Bacteriological studies on limburger cheese. Ph.D. thesis. Agricultural University, Wageningen, The Netherlands.

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