Application of a Loop-Mediated Isothermal Amplification (LAMP) Assay for the Detection of Listeria monocytogenes in Cooked Ham.

Changing eating habits and rising demand of food have increased the incidence of foodborne diseases, particularly in industrialized countries. In this context, contaminated ready-to-eat food (RTE) may be a vehicle for the transmission of Listeria monocytogenes (L. monocytogenes), a foodborne pathogen responsible of listeriosis, a severe infectious disease involving humans and animals. It would be useful to have rapid detection methods to screen the presence of L. monocytogenes in food. In this study, a colorimetric Loop-mediated isothermal amplification (LAMP) assay was applied to the detection of L. monocytogenes in 37 experimentally contaminated RTE meat samples. The LAMP primers consisted of a set of six primers targeting eight regions on the hlyA gene; the assay was carried out in 30 min at 65 °C in a water bath. Amplification products were visualized by color change assessment. The results of colorimetric LAMP assays based on the hly gene obtained in this study were compared to microbiological cultural methods, real-time PCR and real-time LAMP PCR, which show 100% specificity and sensitivity. These data suggest that colorimetric LAMP assays can be used as a screen to detect L. monocytogenes in ready-to-eat meat food.

Phylogenetic and Evolutionary Genomic Analysis of Listeria monocytogenes Clinical Strains in the Framework of Foodborne Listeriosis Risk Assessment.

Listeria monocytogenes is one of the most important foodborne pathogens responsible for listeriosis, a severe disease with symptoms ranging from septicemia, meningoencephalitis, and abortion. Given the strong impact of listeriosis on human health and the difficulty of controlling L. monocytogenes along the food production chain, listeriosis has become a priority subjected to molecular surveillance in European Union/European Economic Area since 2007. From 2018, surveillance is based on whole-genome sequence using the core genome multilocus sequence type. The complete sequences of 132 clinical strains were used to define the evolutionary relatedness among subtypes of L. monocytogenes isolated in Italy from 2010 to 2016, allowing the identification of clades and/or clusters associated with outbreaks or sporadic cases of listeriosis. All the strains analyzed are clustered in lineages I and II, and the majority of the strains were classified as lineage II. A probable epidemic entrance in different years for every clade and cluster from each different region was defined. The persistence of the same specific clonal complexes of L. monocytogenes has been found over long periods; this may be related to the fact that some strains are able to survive better than others in a food production environment. Phylogenic studies, using whole-genome sequence data, are able to identify the emergence of highly persistent pathogenic variants, contributing to improving the hazard characterization of L. monocytogenes.

Quality of ready-to-eat swordfish fillets inoculated with Lactobacillus paracasei IMPC 2.1.

BACKGROUND: The quality of marinated ready-to-eat (RTE) swordfish fillets, with or without inoculation with the probiotic strain Lactobacillus paracasei IMPC 2.1, was assessed over 3 months of refrigerated storage at 4 °C. RTE probiotic and control fish fillets were sampled after 7, 14, 30, 60, and 90 days of storage. Microbiological tests were performed, and fatty acid (FA) profiles and malondialdehyde content were examined. Microbiological counts, including total viable count, lactic acid bacteria (LAB), yeasts, moulds, Enterobacteriaceae, and Pseudomonadaceae were determined. RESULTS: Inoculation successfully ensured the growth of the probiotic strain and prevented the growth of other LAB. The two RTE products showed significant differences in lipid profile and lipid oxidation during storage. In particular, inoculation with L. paracasei IMPC 2.1 increased the amount of polyunsaturated FAs and limited the amount of monounsaturated FAs and oleic acid, as well as lipid oxidation. It thus represents an interesting strategy for preserving the chemical quality of fish fillets and an alternative means of delivering probiotics. CONCLUSION: Probiotic inoculation with Lactobacillus paracasei seemed to delay lipid oxidation of the fish flesh and increase the retention of polyunsaturated FAs, suggesting a potential application for this strain in the seafood industry. © 2018 Society of Chemical Industry.

Dissemination and Persistence of Pseudomonas spp. in Small-Scale Dairy Farms.

This study was aimed at collecting data on presence, dissemination and persistence of Pseudomonas in small-scale dairy farms. Six farms (located in Piedmont) were visited three times over 2014: 116 waters (wells and different faucets/pipes) and 117 environmental samples (milking equipments and drains) were collected. Enumeration of Pseudomonadaceae was performed, 3-5 colonies/samples were selected for identification via 16SrDNA/oprI polymerase chain reaction (PCR), and typed by enterobacterial-repetitive-intergenic-consensus (ERIC)-PCR. Pseudomonadaceae were detected in 77% of samples. No statistical differences were found among proportions of positives across farms, sample typologies and seasons. Most isolates were Pseudomonas fluorescens (45%), and ERIC-PCR showed 32 persistent types diffused across farms. All in all, Pseudomonas spp. represents a challenge, considering its presence over time in water as well as in teat cups, indicating a continuous source of contamination. Moreover, persistency of strains may indicate biofilm-formation and/or sanitisers resistance, therefore emphasising the role of primary production for preventing milk contamination by Pseudomonas spp.

Effects of Rosemary Oil (Rosmarinus officinalis) on the Shelf-Life of Minced Rainbow Trout (Oncorhynchus mykiss) during Refrigerated Storage.

The effects of three concentrations (0.2%, 1% and 3%) of rosemary oil (RO) on the freshness indicators, oxidative stability, fatty acid and biogenic amine (BA) contents of minced rainbow trout muscle (MTM) were investigated after different periods of storage (three and nine days) at 4 ± 1 °C. Moreover, the terpene and sesquiterpene contents in the treated MTM were also measured. RO treatment improves the pH, oxidative stability of the lipids and the FA profile, which resulted in a significant extension of MTM shelf-life. Storage time influenced all freshness indicators, with the exception of yellowness and chroma. Treatment with RO had a positive effect, leading to low BA content, especially putrescine, cadaverine, tyramine and histamine. Differences in BA were also found to be due to storage time, with the exception of spermidine, which was not influenced by time. Moreover, the presence of the terpenoid fraction of RO in MTM improved the quality of this ready-to-cook fish food.