In recent years, there has been an increase in the consumption of seafood such as shellfish and crustaceans due to their pleasant taste and nutritional value. Fish are also a crucial part of a healthy, balanced diet. However, the consumption of these products may cause food poisoning through marine biotoxins. In recent years, several legal acts have been published by the European Commission to regulate toxin limits and describe their reference analysis methods. Commission Regulation (EC) No. 853/2004 established the maximum contents of marine biotoxins only in bivalve mollusks. Although other groups of marine organisms such as crustaceans (crabs, shrimps, and lobsters) and fish are not included in the EU rules for toxin monitoring, they may still be vectors of marine biotoxins for humans. Due to this, there is an urgent need for studies regarding the occurrence of marine biotoxins in non-bivalve seafood organisms and their potential influence on public health. In this review, the most important cases of accumulation of marine biotoxins in crustaceans and fish in recent years are described.

The rapid growth of world population and increase in living standards have led to an increase in the demand for high-quality, safe food. The Food and Agriculture Organization of the United Nations (FAO) estimates that by 2050 the demand for food will increase by 60%, and production of animal protein will increase by 1.7% a year, with meat production to increase by nearly 70%, dairy products by 55%, and aquaculture by as much as 90%. Microbial contamination of food is a significant problem for the accessibility of safe food which does not pose a threat to the life and health of consumers. Campylobacter, Salmonella, and Yersinia are responsible for thousands of food-borne infections in humans. Currently, numerous programs are being developed to combat pathogenic bacteria in the food supply chain, especially at the primary production stage. These approaches include physical, chemical, biological, and other strategies and methods used to inhibit the bacterial growth of bacteria or completely eliminate the pathogens from the food chain. Therefore, an extremely important goal is to provide safe food and control its quality by eliminating pathogenic and spoilage microorganisms. However, the use of chemicals in food preservation has negative effects for both the consumption values of food and the natural environment. Therefore, it seems absolutely necessary to implement measures utilizing the most environmentally friendly and effective techniques for controlling microbial contamination in food. There is a great need to develop ecological methods in food production which guarantee adequate safety. One of these methods is the use of bacteriophages (bacterial viruses) naturally occurring in the environment. Given the above, the aim of this study was to present the most natural, ecological, and alternative methods of food preservation with regard to the most common foodborne zoonotic bacteria. We also present methods for reducing the occurrence of microbial contamination in food, thus to produce maximally safe food for consumers.

Biogenic amines (BA) are simple organic bases of low molecular weight, formed during decarboxylation of amino acids. Ripened cheeses provide suitable conditions for the development of bacteria and production of BAs. The aim of the present study was to investigate the presence of eight BAs in ripened cheese samples (n = 125) using a high-performance liquid chromatography with diode array detector (HPLC-DAD). Furthermore, microbiological analyses towards identification of bacteria using matrix-assisted laser desorption ionisation—time of flight mass spectrometry (MALDI-TOF MS) were performed. Cadaverine and putrescine were detected in 28.0% and 20.8% of cheese samples at concentrations ranging from 6.12 to 2871 mg/kg and 5.74 to 441 mg/kg, respectively. High amounts of putrescine and cadaverine in cheeses were associated with the presence of Hafnia alvei. Tyramine was identified in 28.0% of samples in the concentration range of 5.62–646 mg/kg. High concentrations of this amine was found in cheeses containing Enterococcus faecium and Enterococcus faecalis. Histamine content, the only BA restricted in food according to Regulation 2073/2005, was observed above 100 mg/kg in 11.2% of the cheeses. Ripened cheeses available on the local retail market may contain significant levels of biogenic amines and may pose a potential health hazard to consumers.

The presence of Shiga toxin-producing Escherichia coli (STEC) on bovine and porcine carcasses during 2019-2023 was investigated. A total of 368 bovine and 87 porcine carcasses were tested using the ISO/TS 13136 standard and the STEC isolates were further characterized with whole genomic sequencing (WGS). It was found that 119 (32.3 %) of bovine and 14 (16.1 %) of porcine carcasses were positive for the stx Shiga toxin gene. Further analysis of the stx-positive samples allowed to isolate 32 (26.9 %) bovine and two (14.3 %) porcine STEC, respectively. Bovine isolates were classified into 21 different serotypes with the most prevalent O168:H8 (3 isolates), whereas two porcine STEC belonged to two serotypes that were not identified in bovine strains. Isolates of bovine carcass origin were mainly positive for the stx2 Shiga toxin gene, either alone or in combination with stx1 type (26 of 32; 81.2 % isolates). Two STEC from porcine carcasses were positive for the stx2e variant only. All STEC, irrespective of the origin, were negative for the eae intimin gene. MLST and cgMLST analyses of all strains tested revealed that they were diverse. However, a close molecular relationship between some bovine isolates based on cgMLST schemes was observed. Comparison of the current bovine STEC with those isolated between 2014 and 2018 showed that some of them consisted of the same MLST sequence types. However, based on cgMLST analysis only two cases of three genomes of STEC isolates each (two from period 2019-2023 and one isolated between 2014 and 2018) revealed up to 50 allelic differences.

The consumption of organic food has increased in recent years. In organic rearing animals are exposed to outdoor conditions, which may increase their risk of infection from various pathogens. In the present study the occurrence of the most significant foodborne pathogenic bacteria in organic meat and ready-to-eat organic meat products was assessed. Out of 100 raw organic meat samples tested, 72 were contaminated with bacterial pathogens. The highest percentage of contaminated samples was observed in poultry meat (92.5%) followed by pork meat (66.7%). Furthermore, 50.0% of beef origin samples were positive for the bacteria tested. L. monocytogenes was found in 39.0% of samples, S. aureus was identified in 37.0%, Campylobacter in 20.0%, Salmonella in 8.0% and Shigatoxin-producing E. coli in 4.0% of raw meat samples. In 31.0% of samples a co-occurrence of two (83.9%) or three (16.1%) pathogens was observed. Among 100 samples of organic meat products tested, only L. monocytogenes was found in 5.0% of samples. The result of the present study indicated that organic food may be a source of harmful microorganisms that may pose foodborne infections to consumers.

Listeria monocytogenes is an important food-borne pathogen that causes severe disease in humans. Many different foods products may be vehicles of these pathogenic strains. The aims of this study were to identified serogroups L. monocytogenes isolates by using PCR and analyze the distribution of serogroups in different food products of animal origin. L. monocytogenes isolates from ready-to-eat (RTE) meat products (n = 1,571), non-meat RTE food (n = 237), and non-RTE products (n = 189) were collected during years 2013-2020. A total of 899 (45.0%) isolates, irrespective of the origin, were classified to serogroup IIa. The remaining 1,098 strains belonged either to serogroups IVb (478; 23.9%), IIc (347; 17.4%) or IIb (273; 13.7%), respectively. In the predominant group products, i.e. RTE food of meat origin, serogroup IIa (702; 44.7%) was the most common followed by IVb (334; 21.3%), IIc (300; 19.1%), and IIb (235; 14.9%), respectively. The statistical differences were observed between the prevalence of L. monocytogenes serogroup IIb in sausages and cold cuts (P < 0.0001), cold cuts and delicatessen food (P < 0.001), as well as strains of serogroup IVb from sausages and cold cuts (P < 0.001). The prevalence of serogroup IIa was statistically higher among strains of milk and milk products origin than RTE fish and fish products (P < 0.05). The differences (P < 0.0001) were also observed in the distribution of L. monocytogenes of serogroup IIa from mixed food and milk as well as fish products. Additionally, the occurrence of serogroup IVb in mixed food differed compared to other food categories, i.e. milk and fish products (P < 0.0001). The obtained results confirmed the presence of L. monocytogenes serogroups IIa and IVb in food of animal origin, including RTE products, that are responsible for listeriosis in humans and suggested that these food products may pose a potential health problem, particularly to sensitive groups of people.

Abstract 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.

Introduction Campylobacteriosis is the most common human foodborne bacterial infection worldwide and is caused by bacteria of the Camplylobacter genus. The main source of these bacteria is poultry, but other food-producing animals such as pigs are also responsible for human infections. An increasing number of strains with resistance to fluoroquinolones and other antimicrobials such as macrolides were recently noted. The aim of the study was to investigate Campylobacter contamination of porcine carcasses and determine the antimicrobial resistance of the obtained isolates.Material and Methods A total of 534 swabs from carcasses of pigs slaughtered in Poland during 2019-2022 were tested for Campylobacter spp. Results Campylobacter was detected in 164 (30.7%) carcasses; among them 149 (90.8%) were classified as C. coli and the remaining 15 (9.2%) samples were C. jejuni-positive. Because a low number of C. jejuni isolates were identified, only the C. coli isolates were subjected to antimicrobial resistance analysis. The majority of these isolates were resistant to streptomycin (94.0%), ciprofloxacin (65.8%) and tetracycline (65.1%). A total of 94 (63.1%) strains displayed antimicrobial multiresistance patterns and were mainly resistant to fluoroquinolones, aminoglycosides and tetracyclines (74; 49.7% of the isolates tested). Conclusion The obtained results showed that pig carcasses may be contaminated with antimicrobial-resistant C. coli.

Listeria monocytogenes is one of the most dangerous food-borne pathogens and is responsible for human listeriosis, a severe disease with a high mortality rate, especially among the elderly, pregnant women and newborns. Therefore, this bacterium has an important impact on food safety and public health. It is able to survive and even grow in a temperature range from -0.4°C to 45°C, a broad pH range from 4.6 to 9.5 and at a relatively low water activity (aW < 0.90), and tolerates salt content up to 20%. It is also resistant to ultraviolet light, biocides and heavy metals and forms biofilm structures on a variety of surfaces in food-production environments. These features make it difficult to remove and allow it to persist for a long time, increasing the risk of contamination of food-production facilities and ultimately of food. In the present review, the key mechanisms of the pathogen's survival and stress adaptation have been presented. This information may grant better understanding of bacterial adaptation to food environmental conditions.

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.

The aim of this study was to evaluate the microbiological contamination of raw bivalve molluscan shellfish (BMS) available on the Polish market and determinate the antimicrobial resistance of the obtained isolates. A total of 1000 mollusc samples were tested for the presence of Salmonella spp., L. monocytogenes, V. parahaemolyticus, and S. aureus using the ISO standard methods. Additionally, the bacterial isolates’ susceptibility to antimicrobials was determined using the minimum inhibitory concentration (MIC) method. The obtained results showed that Salmonella spp. was detected in 31 (3.1%) samples, and 51.6% of the bacterial isolates were classified as Salmonella Typhimurium. A total of 74.2% of the Salmonella isolates were sensitive to all antimicrobial agents, whereas three isolates were multiresistant. L. monocytogenes was isolated from 18 (1.8%) BMS, and the isolates belonged to serogroups IIa, IIb, and IVb. Most of them were resistant to ceftriaxone (77.8%) and oxacillin (55.6%). V. parahaemolyticus was present in 24.2% BMS. These isolates were mainly resistant to ampicillin (77.3%) and streptomycin (64.0%). Moreover, 15.2% of the bivalve molluscs were contaminated with S. aureus. Most isolates belonging to this species were resistant to penicillin (84.9%). A total of 60 (6.0%) bivalve molluscs were contaminated with more than one pathogen simultaneously. In addition, the tested bacteria were more likely to be identified during the warmer period (53.9%) compared to the samples analyzed in colder months (35.7%). The obtained results indicate that raw bivalve molluscs from the Polish market are frequently contaminated with bacterial foodborne pathogens, which may be resistant to antimicrobials.

Listeriosis is a serious food-borne illness, especially in susceptible populations, including children, pregnant women, and elderlies. The disease can occur in two forms: non-invasive febrile gastroenteritis and severe invasive listeriosis with septicemia, meningoencephalitis, perinatal infections, and abortion. Expression of each symptom depends on various bacterial virulence factors, immunological status of the infected person, and the number of ingested bacteria. Internalins, mainly InlA and InlB, invasins (invasin A, LAP), and other surface adhesion proteins (InlP1, InlP4) are responsible for epithelial cell binding, whereas internalin C (InlC) and actin assembly-inducing protein (ActA) are involved in cell-to-cell bacterial spread. L. monocytogenes is able to disseminate through the blood and invade diverse host organs. In persons with impaired immunity, the elderly, and pregnant women, the pathogen can also cross the blood–brain and placental barriers, which results in the invasion of the central nervous system and fetus infection, respectively. The aim of this comprehensive review is to summarize the current knowledge on the epidemiology of listeriosis and L. monocytogenes virulence mechanisms that are involved in host infection, with a special focus on their molecular and cellular aspects. We believe that all this information is crucial for a better understanding of the pathogenesis of L. monocytogenes infection.

Introduction Histamine intoxication, known as scombroid fish poisoning, is caused by the consumption of foods with high levels of histamine. This biogenic amine is formed as a result of histidine decarboxylation by bacterial decarboxylases present in food, including fish and fish products. The aim of this study was to investigate the content of histamine at different production stages of canned, marinated and smoked fish. Material and Methods Raw fish, semi-finished fish products, and the final products of the same production batches were collected between 2019 and 2022 from different fish production facilities in Poland. A total of 133 raw fish samples and 76 smoked fish, 54 brined fish, 39 canned fish and 18 marinated fish final products were analysed using high performance liquid chromatography with a diode array detector. Results Histamine was identified in 55 (17.2%) out of 320 tested samples, including 8 samples of raw fish with a histamine level above 100 mg/kg. However, no samples of fish products had histamine content above the European Union Commission limit. Conclusion The obtained results show that fish products on the Polish market are generally safe for consumers in regard to histamine intoxication risk.

Background: A global problem of multi-drug resistance (MDR) among bacteria is the cause of hundreds of thousands of deaths every year. In response to the significant increase of MDR bacteria, legislative measures have widely been taken to limit or eliminate the use of antibiotics, including in the form of feed additives for livestock, but also in metaphylaxis and its treatment, which was the subject of EU Regulation in 2019/6. Numerous studies have documented that bacteria use both phenotypis and gentic strategies enabling a natural defence against antibiotics and the induction of mechanisms in increasing resistance to the used antibacterial chemicals. The mechanisms presented in this review developed by the bacteria have a significant impact on reducing the ability to combat bacterial infections in humans and animals. Moreover, the high prevalence of multi-resistant strains in the environment and the ease of transmission of drug-resistance genes between the different bacterial species including commensal flora and pathogenic like foodborne pathogens (E. coli, Campylobacter spp., Enterococcus spp., Salmonella spp., Listeria spp., Staphylococcus spp.) favor the rapid spread of multi-resistance among bacteria in humans and animals. Given the global threat posed by the widespread phenomenon of multi-drug resistance among bacteria which are dangerous for humans and animals, the subject of this study is the presentation of the mechanisms of resistance in most frequent bacteria called as “foodborne pathoges” isolated from human and animals. In order to present the significance of the global problem related to multi-drug resistance among selected pathogens, especially those danger to humans, the publication also presents statistical data on the percentage range of occurrence of drug resistance among selected bacteria in various regions of the world. In addition to the phenotypic characteristics of pathogen resistance, this review also presents detailed information on the detection of drug resistance genes for specific groups of antibiotics. It should be emphasized that the manuscript also presents the results of own research i.e., Campylobacter spp., E. coli or Enetrococcus spp. This subject and the presentation of data on the risks of drug resistance among bacteria will contribute to initiating research in implementing the prevention of drug resistance and the development of alternatives for antimicrobials methods of controlling bacteria.

Biogenic amines (BAs) are organic, basic nitrogenous compounds formed during the decarboxylation of amino acids. A method for the determination of eight biogenic amines (tryptamine, 2-phenyletylamine, putrescine, cadaverine, histamine, tyramine, spermidine, spermine) in ripened cheeses was developed and validated. Cheese samples with the addition of internal standards were extracted with 0.2 M perchloric acid and pre-column derivatized with dansyl chloride at 60 °C for 15 min, purified with toluene and dried under a stream of nitrogen. The samples were analyzed using high performance liquid chromatography with diode array detector (HPLC-DAD). The method was validated with the BAs at three concentration levels: 50, 100, and 200 mg/kg, respectively. The obtained values of correlation coefficient (R2) ranged at 0.9997–0.9998 for all of compounds. The limits of detection (LOD) and quantification (LOQ) were in ranges 1.53–1.88 and 5.13–6.28 mg/kg, respectively. The recovery for all of biogenic amines ranged from 70 to 120% and the precision (RSDr) value were <20%. The validated method was applied to analysis of 35 real ripened cheese samples purchased in Poland.

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.

Abstract 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.

INTRODUCTION:Marine biotoxins are toxic substances that may cause illness and death in marine organisms and humans. This article disseminates the results of a 4-year study on the occurrence of marine biotoxins in raw bivalve molluscs purchased from Polish suppliers. MATERIAL AND METHODS:A total of 256 samples of 8 different molluscs species were analysed for the presence of biotoxins using the ELISA method for paralytic shellfish poison, diarrhoetic shellfish poison, and amnaesic shellfish poison. RESULTS:The permitted limits of marine biotoxin content were not exceeded in any of the analysed samples and the majority of them were free from these compounds. CONCLUSION:The results of the study indicate that the tested raw bivalve molluscs available in Poland were safe for consumers.

The organic food sector and consumer interest in organic products are growing continuously. The safety and quality of such products must be at least equal to those of conventional equivalents, but attaining the same standards requires overcoming a particular problem identified in organic food production systems: the occurrence of bacterial pathogens such as Salmonella, Campylobacter, Listeria monocytogenes, Staphylococcus aureus and pathogenic Escherichia coli. These food-borne microorganisms were detected in the production environments of such food. The prevalence of pathogenic bacteria in organic livestock and products may be higher, but may also be the same as or lower than in like material from conventional farms. Furthermore, the incidence of antimicrobial-resistant bacteria was more often detected in conventional than in organic production. The aim of this review was to present the recent information on the microbiological safety of food of animal origin produced from raw materials from organic farms.

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.