Journal
JOURNAL OF DAIRY SCIENCE
Volume 96, Issue 8, Pages 4928-4937Publisher
ELSEVIER SCIENCE INC
DOI: 10.3168/jds.2013-6688
Keywords
high-throughput DNA sequencing; flow cytometry; quantitative PCR
Funding
- FP7 EFRAIM FP7-KBBE-2007-1-'Mechanisms of early protective exposures on allergy development'
- Department of Agriculture, Fisheries and Food (Dublin, Ireland)
- Teagasc Walsh Fellowships
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The microbial composition of raw and pasteurized milk is assessed on a daily basis. However, many such tests are culture-dependent, and, thus, bacteria that are present at subdominant levels, or that cannot be easily grown in the laboratory, may be overlooked. To address this potential bias, we have used several culture-independent techniques, including flow cytometry, real-time quantitative PCR, and high-throughput DNA sequencing, to assess the microbial population of milk from a selection of commercial milk producers, pre- and postpasteurization. The combination of techniques employed reveals the presence of a previously unrecognized and diverse bacterial population in unpasteurized cow milk. Most notably, the use of high-throughput DNA sequencing resulted in several bacterial genera being identified in milk samples for the first time. These included Bacteroides, Faecalibacterium, Prevotella, and Catenibacterium. Our culture-independent analyses also indicate that the bacterial population of pasteurized milk is more diverse than previously appreciated, and that nonthermoduric bacteria within these populations are likely to be in a damaged, nonculturable form. It is thus apparent that the application of state-of-the-art approaches can provide a detailed insight into the bacterial composition of milk and could potentially be employed in the future to investigate the factors that influence the composition of these populations.
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