Microbial shifts through the ripening of the "Entre Serras" Minas artisanal cheese monitored by high-throughput sequencing

Abstract

Minas Gerais is a Brazilian state known as the largest cheese producer in Brazil. Minas Artisanal Cheese (MAC) is produced in different regions of this Brazilian state using raw cow milk to which a natural starter culture ("pingo") is added. "Entre Serras" is one of these regions, in which the MAC production had decreased (even stopped) for decades until recently, when artisanal cheeses production has been resurrected. Here, we aimed to gain insights on the bacterial diversity of "Entre Serras" MAC. 16S rRNA gene amplicon sequencing was used to assess the bacterial community in cheeses produced by four farms (A, B, C, and D) over 60 days of ripening. Overall, Lactococcus lactis was the predominant species found, regardless of the producer/farm. Enterococcus, Streptococcus, Lactobacillus and Leuconostoc genera were also prevalent in the samples microbiota and their levels varied according to the producer/farm. Cheeses produced by Farms A and B presented high contaminant levels (mainly Enterobacteriaceae and S. aureus), which may be attributed to poor hygiene during cheese production and/or herd health management. Chao1 indices varied significantly when the estimated species richness values of the producers/farms were compared (p < 0.05). A principal coordinate analysis also revealed distinct microbial communities for some farms (p < 0.001). However, no statistical significance was identified when samples were grouped by ripening time. Core microbiota analysis indicated that "Entre Serras" MAC microbiota includes not only LAB, but also spoilage and potentially pathogenic bacteria. We provide the first insights on the bacterial diversity of "Entre Serras" MAC, helping the understanding of the inter-regional microbiological diversity of the samples.

Keywords: 16S rRNA; Artisanal cheese; Lactic acid bacteria; Microbiome.