RESUMO
Reported cases of listeriosis from food of non-animal origin (FNAO) are increasing. In order to assess the risk of exposure to Listeria monocytogenes from FNAO, the genetic characterization of the pathogen in FNAO products and in primary production and processing plants needs to be investigated. For this, 123 samples of fresh and frozen soft fruit and 407 samples of 39 plants in Bavaria, Germany that produce and process FNAO were investigated for Listeria contamination. As a result, 64 Listeria spp. isolates were detected using ISO 11290-1:2017. Environmental swabs and water and food samples were investigated. L. seeligeri (36/64, 56.25%) was the most frequently identified species, followed by L. monocytogenes (8/64, 12.50%), L. innocua (8/64, 12.50%), L. ivanovii (6/64, 9.38%), L. newyorkensis (5/64, 7.81%), and L. grayi (1/64, 1.56%). Those isolates were subsequently sequenced by whole-genome sequencing and subjected to pangenome analysis to retrieve data on the genotype, serotype, antimicrobial resistance (AMR), and virulence markers. Eight out of sixty-four Listeria spp. isolates were identified as L. monocytogenes. The serogroup analysis detected that 62.5% of the L. monocytogenes isolates belonged to serogroup IIa (1/2a and 3a) and 37.5% to serogroup IVb (4b, 4d, and 4e). Furthermore, the MLST (multilocus sequence typing) analysis of the eight detected L. monocytogenes isolates identified seven different sequence types (STs) and clonal complexes (CCs), i.e., ST1/CC1, ST2/CC2, ST6/CC6, ST7/CC7, ST21/CC21, ST504/CC475, and ST1413/CC739. The core genome MLST analysis also showed high allelic differences and suggests plant-specific isolates. Regarding the AMR, we detected phenotypic resistance against benzylpenicillin, fosfomycin, and moxifloxacin in all eight L. monocytogenes isolates. Moreover, virulence factors, such as prfA, hly, plcA, plcB, hpt, actA, inlA, inlB, and mpl, were identified in pathogenic and nonpathogenic Listeria species. The significance of L. monocytogenes in FNAO is growing and should receive increasing levels of attention.
RESUMO
Several foodborne outbreaks associated with food of non-animal origin (FNAO) were reported within the last years. In recent years, Listeriamonocytogenes has been associated with such outbreaks. For this reason, different producers of FNAO at the primary production and processing level in Bavaria, Germany, were inspected from July 2020 to June 2021. Environmental and food sampling as well as the sampling of irrigation and processing water was performed to investigate the prevalence of Listeriaspp., including L.monocytogenes at facilities that produce ready-to-eat FNAO. Altogether, 39 producers of soft fruit, vegetables, ready-to-eat raw fruits, and vegetables/fresh cut were inspected. In addition to the on-spot inspections, 407 samples were taken in total, among them, 229 were swab samples from food contact material and the environment, 59 food samples (including soft fruit, vegetables and ready-to-eat vegetables), and 119 samples of irrigation and processing water. Samples were analyzed using methods according to ISO11290-1:2017. Furthermore, the samples of irrigation and processing water were also quantitatively tested for the number of Escherichiacoli (ISO9308-2:2014-06), enterococci (ISO7899-2:2000-11), and Pseudomonasaeruginosa (ISO16266:2008-05). No contamination with E.coli, enterococci, and P.aeruginosa could be detected in most of the samples. Overall, in 12.53% of the samples, Listeriaspp. were detected. L.monocytogenes was identified in 1.72% of the environmental and processing water samples, whereas L.monocytogenes was not detected in food samples. In addition to water sources and quality, this study demonstrates that irrigation regime, cultivation, hygienic handling, and maintenance protocols are highly important to reduce the potential contamination of ready-to-eat soft fruits and vegetables with Listeriaspp.
