Antimicrobial resistance of Listeria monocytogenes serogroups IIa and IVb from food and food-production environments in Poland.

Introduction: Listeria monocytogenes is an important foodborne pathogen responsible for human listeriosis, which is a disease with high hospitalisation and mortality rates. The bacteria are usually susceptible to most antibacterial substances, but resistance to some of them has been recently observed. The present study introduces the evidence on the emergence of antibiotic resistance among L. monocytogenes strains isolated from food and food-production environments in Poland. Material and Methods: A total of 283 L. monocytogenes isolates classified into serogroups IIa and IVb which had been recovered from food and food production environments were tested with 17 antimicrobials. These included those that are recommended for treatment of severe listeriosis cases in humans. A multiplex PCR was used to identify serogroups, and a microbroth dilution method was applied for the determination of antibiotic resistance among the isolates tested. Results: Only 34 (12.0%) strains were susceptible to all the antimicrobials used in the study. The remaining 249 (88.0%) strains displayed different instances of resistance to the antimicrobials tested, from insusceptibility to one (112 strains; 39.6%) to resistance to four antibacterial substances (6 strains; 2.1%). Among them, there were 38 strains (13.4%) with multiresistance patterns. Conclusion: Polish food and its processing environments may be a potential source of antimicrobial-resistant L. monocytogenes, which may pose a potential health risk to consumers in the country.

Listeria monocytogenes in foods-From culture identification to whole-genome characteristics.

Listeria monocytogenes is an important foodborne pathogen, which is able to persist in the food production environments. The presence of these bacteria in different niches makes them a potential threat for public health. In the present review, the current information on the classical and alternative methods used for isolation and identification of L. monocytogenes in food have been described. Although these techniques are usually simple, standardized, inexpensive, and are routinely used in many food testing laboratories, several alternative molecular-based approaches for the bacteria detection in food and food production environments have been developed. They are characterized by the high sample throughput, a short time of analysis, and cost-effectiveness. However, these methods are important for the routine testing toward the presence and number of L. monocytogenes, but are not suitable for characteristics and typing of the bacterial isolates, which are crucial in the study of listeriosis infections. For these purposes, novel approaches, with a high discriminatory power to genetically distinguish the strains during epidemiological studies, have been developed, e.g., whole-genome sequence-based techniques such as NGS which provide an opportunity to perform comparison between strains of the same species. In the present review, we have shown a short description of the principles of microbiological, alternative, and modern methods of detection of L. monocytogenes in foods and characterization of the isolates for epidemiological purposes. According to our knowledge, similar comprehensive papers on such subject have not been recently published, and we hope that the current review may be interesting for research communities.

Listeria monocytogenes - How This Pathogen Survives in Food-Production Environments?

The foodborne pathogen Listeria monocytogenes is the causative agent of human listeriosis, a severe disease, especially dangerous for the elderly, pregnant women, and newborns. Although this infection is comparatively rare, it is often associated with a significant mortality rate of 20-30% worldwide. Therefore, this microorganism has an important impact on food safety. L. monocytogenes can adapt, survive and even grow over a wide range of food production environmental stress conditions such as temperatures, low and high pH, high salt concentration, ultraviolet lights, presence of biocides and heavy metals. Furthermore, this bacterium is also able to form biofilm structures on a variety of surfaces in food production environments which makes it difficult to remove and allows it to persist for a long time. This increases the risk of contamination of food production facilities and finally foods. The present review focuses on the key issues related to the molecular mechanisms of the pathogen survival and adaptation to adverse environmental conditions. Knowledge and understanding of the L. monocytogenes adaptation approaches to environmental stress factors will have a significant influence on the development of new, efficient, and cost-effective methods of the pathogen control in the food industry, which is critical to ensure food production safety.

Genetic Diversity and Relationships of Listeria monocytogenes Serogroup IIa Isolated in Poland.

In the present study, 100 L. monocytogenes isolates of serogroup IIa from food and food production environments in Poland were characterized towards the presence of virulence, resistance, and stress response genes using whole-genome sequencing (WGS). The strains were also molecularly typed and compared with multi-locus sequence typing (MLST) and core genome MLST analyses. The present isolates were grouped into 6 sublineages (SLs), with the most prevalent SL155 (33 isolates), SL121 (32 isolates), and SL8 (28 isolates) and classified into six clonal complexes, with the most prevalent CC155 (33 strains), CC121 (32 isolates), and CC8 (28 strains). Furthermore, the strains were grouped to eight sequence types, with the most prevalent ST155 (33 strains), ST121 (30 isolates), and ST8 (28; strains) followed by 60 cgMLST types (CTs). WGS data showed the presence of several virulence genes or putative molecular markers playing a role in pathogenesis of listeriosis and involved in survival of L. monocytogenes in adverse environmental conditions. Some of the present strains were molecularly closely related to L. monocytogenes previously isolated in Poland. The results of the study showed that food and food production environments may be a source of L. monocytogenes of serogroup IIa with pathogenic potential.

Molecular Typing of Listeria monocytogenes IVb Serogroup Isolated from Food and Food Production Environments in Poland.

Listeria monocytogenes is one of the most important foodborne pathogens that may be present in food and in food processing environments. In the present study, 91 L. monocytogenes isolates of serogroup IVb from raw meat, ready-to-eat food and food production environments in Poland were characterized by whole genome sequencing (WGS). The strains were also compared, using core genome multi-locus sequence typing (cgMLST) analysis, with 186 genomes of L. monocytogenes recovered worldwide from food, environments, and from humans with listeriosis. The L. monocytogenes examined belonged to three MLST clonal complexes: CC1 (10; 11.0% isolates), CC2 (70; 76.9%), and CC6 (11; 12.1%). CC1 comprised of two STs (ST1 and ST515) which could be divided into five cgMLST, CC2 covered two STs (ST2 and ST145) with a total of 20 cgMLST types, whereas CC6 consisted of only one ST (ST6) classified as one cgMLST. WGS sequences of the tested strains revealed that they had several pathogenic markers making them potentially hazardous for public health. Molecular comparison of L. monocytogenes strains tested in the present study with those isolated from food and human listeriosis showed a relationship between the isolates from Poland, but not from other countries.

Antimicrobial resistance and genotypic characteristics of Listeria monocytogenes isolated from food in Poland.

The aim of the study was to determine antimicrobial resistance and genotypic characteristics of L. monocytogenes isolated from food of animal origin from different parts of Poland during years 2013-2016. A total of 146 isolates were tested using a microbroth dilution method, whereas virulence genes and molecular serogroups were identified by PCR. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) methods were used to analyze the genotypic relationship of the strains. Altogether, 102 pulsotypes grouped into 7 clusters and 24 sequence types, including 3 new types, were identified. Most of the strains clustered into individual patterns were originated from different food products and were isolated in different geographical regions at various time. L. monocytogenes was mostly resistant to oxacilin (90.4% strains), clindamycin (54.1%) and ceftriaxone (49.3%). A multiresistance patterns, mainly to ceftriaxone, oxacillin together with other antimicrobials, were observed among 27.4% strains. Antimicrobial resistance and presence of virulence genes suggest that food of animal origin contaminated with L. monocytogenes may present a risk for public health.