Dataset Information

Occurrence of Listeria monocytogenes in Artisanal Cheeses from Poland and Its Identification by MALDI-TOF MS.

ABSTRACT: Listeria monocytogenes is a foodborne pathogen. A source of infection can be artisanal cheeses. Identification of the Listeria species is important for the protection of public health and the food industry. This study aimed to examine artisanal cheeses for the presence of L. monocytogenes and the effectiveness of the MALDI-TOF MS method in the identification of the L. monocytogenes isolates. A total of 370 samples of artisanal cheeses were examined. L. monocytogenes was found in 23 cheese samples (6.2%). The reliability of L. monocytogenes identification achieved by MALDI-TOF MS was varied, and the vast majority of the isolates (27/32) were identified only to the secure genus, probable species level. This study showed that (i) the occurrence of L. monocytogenes in the artisanal cheeses was at a higher level than that in the other EU countries, (ii) the standard of species identification of L. monocytogenes isolates from artisanal cheeses achieved by MALDI-TOF MS was not satisfactory and (iii) the presence of L. monocytogenes in artisanal cheeses remains a problem with regard to the food safety criterion and a potential public health risk.

SUBMITTER: Pyz-Lukasik R

PROVIDER: S-EPMC8224308 | biostudies-literature | 2021 May

REPOSITORIES: biostudies-literature

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Occurrence of Listeria monocytogenes in Artisanal Cheeses from Poland and Its Identification by MALDI-TOF MS.

Pyz-Łukasik Renata R Gondek Michał M Winiarczyk Dagmara D Michalak Katarzyna K Paszkiewicz Waldemar W Piróg-Komorowska Anna A Policht Agata A Ziomek Monika M

Pathogens (Basel, Switzerland) 20210521 6

Listeria monocytogenes is a foodborne pathogen. A source of infection can be artisanal cheeses. Identification of the Listeria species is important for the protection of public health and the food industry. This study aimed to examine artisanal cheeses for the presence of L. monocytogenes and the effectiveness of the MALDI-TOF MS method in the identification of the L. monocytogenes isolates. A total of 370 samples of artisanal cheeses were examined. L. monocytogene ...[more]

PMID: 34064133

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Project description:BackgroundIn the last decade, an innovative approach has emerged for arthropod identification based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Increasing interest in applying the original technique for arthropod identification has led to the development of a variety of procedures for sample preparation and selection of body parts, among others. However, the absence of a consensual strategy hampers direct inter-study comparisons. Moreover, these different procedures are confusing to new users. Establishing optimized procedures and standardized protocols for mosquito identification by MALDI-TOF MS is therefore a necessity, and would notably enable the sharing of reference MS databases. Here, we assess the optimal conditions for mosquito identification using MALDI-TOF MS profiling.MethodsThree homogenization methods, two of which were manual and one automatic, were used on three distinct body parts (legs, thorax, head) of two mosquito laboratory strains, Anopheles coluzzii and Aedes aegypti, and the results evaluated. The reproducibility of MS profiles, identification rate with relevant scores and the suitability of procedures for high-throughput analyses were the main criteria for establishing optimized guidelines. Additionally, the consequences of blood-feeding and geographical origin were evaluated using both laboratory strains and field-collected mosquitoes.ResultsRelevant score values for mosquito identification were obtained for all the three body parts assayed using MALDI-TOF MS profiling; however, the thorax and legs were the most suitable specimens, independently of homogenization method or species. Although the manual homogenization methods were associated with a high rate of identification on the three body parts, this homogenization mode is not adaptable to the processing of a large number of samples. Therefore, the automatic homogenization procedure was selected as the reference homogenization method. Blood-feeding status did not hamper the identification of mosquito species, despite the presence of MS peaks from original blood in the MS profiles of the three body parts tested from both species. Finally, a significant improvement in identification scores was obtained for field-collected specimens when MS spectra of species from the same geographical area were added to the database.ConclusionThe results of the current study establish guidelines for the selection of mosquito anatomic parts and modality of sample preparation (e.g. homogenization) for future specimen identification by MALDI-TOF MS profiling. These standardized operational protocols could be used as references for creating an international MS database.

Project description:BackgroundAnopheles species identification is essential for an effective malaria vector control programme. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) has been developed to identify adult Anopheles species, using the legs or the cephalothorax. The protein repertoire from arthropods can vary according to compartment, but there is no general consensus regarding the anatomic part to be used.MethodsTo determine the body part of the Anopheles mosquitoes best suited for the identification of field specimens, a mass spectral library was generated with head, thorax with wings and legs of Anopheles gambiae, Anopheles arabiensis and Anopheles funestus obtained from reference centres. The MSL was evaluated using two independent panels of 52 and 40 An. gambiae field-collected in Mali and Guinea, respectively. Geographic variability was also tested using the panel from Mali and several databases containing added specimens from Mali and Senegal.ResultsUsing the head and a database without specimens from the same field collection, the proportion of interpretable and correct identifications was significantly higher than using the other body parts at a threshold value of 1.7 (p < 0.0001). The thorax of engorged specimens was negatively impacted by the blood meal after frozen storage. The addition of specimens from Mali into the database significantly improved the results of Mali panel (p < 0.0001), which became comparable between head and legs. With higher identification scores, the using of the head will allow to decrease the number of technical replicates of protein extract per specimen, which represents a significant improvement for routine use of MALDI-TOF MS.ConclusionsThe using of the head of Anopheles may improve the performance of MALDI-TOF MS. Region-specific mass spectrum databases will have to be produced. Further research is needed to improve the standardization in order to share online spectral databases.

Dataset Information

Occurrence of Listeria monocytogenes in Artisanal Cheeses from Poland and Its Identification by MALDI-TOF MS.

Publications

Occurrence of <i>Listeria monocytogenes</i> in Artisanal Cheeses from Poland and Its Identification by MALDI-TOF MS.

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