Environmental monitoring of Listeria monocytogenes contamination in dairy processing facilities combining culturing technique and molecular methods

Abstract

of the present study was to investigate Listeria contamination in Sardinian sheep cheese-making plants using culture and DNA-based methods. Food and environmental samples were collected from different surfaces of 14 facilities. Samples were collected using pre-moistened sponge swabs. Each site was sampled using two sponges used one for microbiological detection and the other for extraction of total DNA. The DNA was submitted to real time PCR targeting the prs gene for Listeria spp., the gene inlA for Listeria monocytogenes and the gene iap for Listeria innocua . DNA metabarcoding was also conducted on the total DNA. Overall were collected 254 environmental and 36 food samples. The prevalence of Listeria spp. and Listeria monocytogenes was 18.6% and 10.0%, respectively. Samples enrichment coupled with quantitative PCR (qPCR) showed a greater sensitivity than the conventional culture method with an overall prevalence of 12.1% for L. monocytogenes . Both, qPCR and ribosomal DNA-metabarcoding conducted on total DNA extracted from the sponges performed poorly in comparison to the culture-based method detecting the presence of Listeria genus in as little as 7 (2.4%) and 4 (1.4%) samples respectively. The present study supports the use of a culture enrichment coupled with qPCR for routine monitoring of contamination in food premises. • Prevalence of Listeria was assessed in Sardinian cheesemaking plants. • Culture-mediated enrichment and DNA-based methods with no enrichment were used. • Metabarcoding was performed on DNA extracted from environmental samples; • The mean prevalence of L. monocytogenes with enrichment methods was 10.0% • Listeria represented as low as 0.001%-0.028% of the total microbial population

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