We addressed this in a variety of ways. First, the extraction kit used to perform the DNA extractions was chosen based on data collected

in which the Qiagen DNeasy Blood and Tissue kit was compared to five other commercially-available kits for the extraction of Brucella neotomae DNA from the same Latin-style cheeses used in this study (T. Lusk, E. Strain, and J.A. Kase, submitted for publication). The Qiagen DNeasy kit was found to produce the highest quality and quantity DNA from this matrix. All extractions were performed by a single person at one time. Lastly, four subsamples of each enriched cheese brand were extracted and sequenced, with all replicates producing Cetuximab molecular weight similar bacterial profiles within each brand except for Brand A, in which 1 replicate showed more diversity than its counterparts. Selleck RG7422 Conclusions This research presents a first look at the microflora of Latin-style cheese using Next-Generation Sequencing. Our findings offer surprising insight into cheese microflora composition, with three cheese brands exhibiting unique bacterial profiles which varied in diversity and abundance of taxa. Although the cheese are visually similar (e.g. white color and soft, crumbly texture), their bacterial profiles were very different at nearly every classification level. Brand A cheese was clearly more diverse than the other two cheese brands

with 13 OTUs at the genus level using a 95% Methocarbamol identity threshold compared to 7 and 3 for Brand C and Brand B, respectively. Additionally, Brand A was dominated by different genus than Brands B and C. Brand B showed less

diversity, mostly dominated at the genus level by Exiguobacterium which constituted 96% of its microflora composition. Exiguobacterium also made up 46% of Brand C’s profile, although its presence in cheese has not been previously documented though it has been found in milk. Factors such as milk, pH, starter culture, and salt concentration may have contributed to the unique bacterial composition of each cheese brand, although no particular factor was determined to be responsible for differences in abundance between the brands based on the limited available information. Overnight enrichment in a non-selective broth also may have allowed some fast-growing bacteria to out-compete and inhibit slower growing bacteria. This emphasizes the importance of examining food samples after the broth enrichment step to provide a more accurate depiction of microflora composition when trying to selectively cultivate target organisms while decreasing competing background flora. More effort is needed to fully characterize cheese microbial populations and to understand the effects of enrichment formulations on population composition. This valuable preliminary data will certainly inform future culture-based efforts.