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In this study, we decided to test the hypothesis that the fungal biostarter M. flavus used during a 21-day beef dry-aging process significantly impacts the composition of other microorganisms, the profile of volatile compounds, meat hardness characteristics, and, consequently, the sensory quality. The experiments were performed on samples derived from animals crossbred between Holstein–Fresian cows and meat breed bulls. Two groups of samples were studied, including the control group, without biostarter, and a group inoculated with the M. flavus biostarter. Both sample groups were seasoned for 21 days in the dry-aging fridge. The physicochemical parameters (pH, color parameters), the chemical composition of muscle, the determination of the shear force, the profile of volatile compounds (VOCs), and the sensory quality were evaluated after aging. During this study, classical microbiological methods were used to investigate the influence of fungal biostarters on the growth and survival of bacteria and other fungi (e.g., yeasts) during the dry-aging process of beef (DAB). The M. flavus biostarter improved the sensory quality of DAB, allowing high sensory quality to be achieved after just 21 days. This is likely due to the diverse VOCs produced by the fungus, including 1-tetradecanol, 2-nonenal, trans-2-undecenoic acid, and the following esters: formic acid hexyl ester, 10-undecenoic acid methyl ester, and 4-methylpentanoic acid methyl ester. The presence of the biostarter had no significant effect on the number of the bacteria or the survivability of the L. monocytogenes on the meat's surface in laboratory conditions. [ABSTRACT FROM AUTHOR]

Bacillus cereus is a foodborne pathogen causing food safety issues due to the formation of difficult to eliminate spores and biofilms. The objective of this study was to investigate the occurrence of B. cereus (conducted as part of monitoring in 2017-2018) and the presence of a toxin gene in strains isolated from retail products (pastries/cakes; vegetables, spices, delicatessen products) in Poland, and to determine the susceptibility of these microorganisms to different antimicrobial agents. A total of 267 B. cereus isolates from food products were examined, of which 95.51% were found positive for the presence of at least one toxin gene, with the highest frequency of the nhe gene (91.39%). The hbl and cytK genes were detected in 53.56% and 44.19% of B. cereus strains, respectively. The lowest frequency was found for the ces gene (2.62%). The susceptibility of B. cereus isolates to 16 antimicrobials was investigated. Ampicillin and penicillin resistance was the most common resistance phenotype and was identified in 100% of the B. cereus isolates. In addition, the tested isolates exhibited resistance to: amoxicillin-clavulanic acid (96.25%), cephalothin (67.79%), ceftriaxone (64.42%), rifampicin (46.82%), trimethoprim-sulfamethoxazole (5.62%), quinupristin/dalfopristin (4.87%), chloramphenicol (3.75%), clindamycin (2.62%), teicoplanin (1.87%), erythromycin (1.87%), ciprofloxacin (0.75%), imipenem (0.75%), tetracycline (0.37%), and gentamicin (0.37%). The study results contribute to characterizing the diversity of B. cereus isolated from various food products in Poland and their impact on food safety and public health. This study delivers practical information on antibiotic resistance and the frequency of toxin genes among strains isolated from food. [ABSTRACT FROM AUTHOR]

The B-cell CLL/lymphoma 11B gene (BCL11B) plays a crucial role in T-cell development, but its role in T-cell malignancies is still unclear. To study its role in the development of T-cell neoplasms, we generated an inducible BCL11B knockout in a murine T cell leukemia/lymphoma model. Mice, bearing human oncogenes TAL BHLH Transcription Factor 1 (TAL1; SCL) or LIM Domain Only 1 (LMO1), responsible for T-cell acute lymphoblastic leukemia (T-ALL) development, were crossed with BCL11B floxed and with CRE-ER/lox mice. The mice with a single oncogene BCL11Bflox/floxCREtg/tgTAL1tg or BCL11Bflox/floxCREtg/tgLMO1tg were healthy, bred normally, and were used to maintain the mice in culture. When crossed with each other, >90% of the double transgenic mice BCL11Bflox/floxCREtg/tgTAL1tgLMO1tg, within 3 to 6 months after birth, spontaneously developed T-cell leukemia/lymphoma. Upon administration of synthetic estrogen (tamoxifen), which binds to the estrogen receptor and activates the Cre recombinase, the BCL11B gene was knocked out by excision of its fourth exon from the genome. The mouse model of inducible BCL11B knockout we generated can be used to study the role of this gene in cancer development and the potential therapeutic effect of BCL11B inhibition in T-cell leukemia and lymphoma. [ABSTRACT FROM AUTHOR]

Abstract Background Bacterial penicillin-binding proteins (PBPs) can be visualized by their ability to bind radiolabeled or fluorescent β-lactam derivatives both whole cells and membrane/cell enriched fractions. Analysis of the Listeria monocytogenes genome sequence predicted ten genes coding for putative PBPs, but not all of their products have been detected in studies using radiolabeled antibiotics, thus hindering their characterization. Here we report the positive identification of the full set of L. monocytogenes PBPs and the characteristics of the hitherto undescribed PBPD2 (Lmo2812). Results Eight L. monocytogenes PBPs were identified by the binding of fluorescent β-lactam antibiotic derivatives Boc-FL, Boc-650 and Amp-Alexa430 to proteins in whole cells or membrane/cell wall extracts. The gene encoding a ninth PBP (Lmo2812) was cloned and expressed in Escherichia coli as a His-tagged protein. The affinity purified recombinant protein had DD-carboxypeptidase activity and preferentially degraded low-molecular-weight substrates. L. monocytogenes mutants lacking the functional Lmo2812 enzyme were constructed and, compared to the wild-type, the cells were longer and slightly curved with bent ends. Protein Lmo1855, previously designated PBPD3, did not bind any of the antibiotic derivatives tested, similarly to the homologous enterococcal protein VanY. Conclusions Nine out of the ten putative L. monocytogenes PBP genes were shown to encode proteins that bind derivatives of β-lactam antibiotics, thus enabling their positive identification. PBPD2 (Lmo2812) was not visualized in whole cell extracts, most probably due to its low abundance, but it was shown to bind Boc-FL after recombinant overexpression and purification. Mutants lacking Lmo2812 and another low molecular mass (LMM) PBP, PBP5 (PBPD1) - both with DD-carboxypeptidase activity - displayed only slight morphological alterations, demonstrating that they are dispensable for cell survival and probably participate in the latter stages of peptidoglycan synthesis. Since Lmo2812 preferentially degrades low-molecular- mass substrates, this may indicate a role in cell wall turnover.

Listeria monocytogenes is a potential hazard for food safety and therefore for public health. The aim of the present study was to determine the prevalence and characteristics of L. monocytogenes in Polish ready-to-eat (RTE) meat products for retail sale. Among the 184,439 food samples collected within the framework of a national official control and monitoring program, only 0.3% were positive for L. monocytogenes. A significant group of products that did not meet the criteria were RTE meat products. This group accounted for 40% of all noncompliant samples. Seventy L. monocytogenes isolates from these RTE meat products (meat, sausages, and delicatessen products with meat) were examined. The majority of the tested isolates (51%) belonged to serogroup 1/2a-3a followed by 1/2c-3c (21%), 1/2b-3b-7 (14%), and 4ab-4b-4d-4e (13%). Serogroup 4a-4c was not present among the tested isolates. All L. monocytogenes isolates harbored the virulence-associated genes inlA, inlC, inlJ, and lmo2672. The llsX marker was detected in 12 (17%) of the 70 isolates. Ampicillin resistance was the most common resistance phenotype and was identified in 83% of the L. monocytogenes isolates. A low incidence of resistance to amoxicillin-clavulanic acid (6% of isolates) was also detected. All L. monocytogenes isolates were susceptible to chloramphenicol, gentamicin, ciprofloxacin, meropenem, sulfamethoxazole-trimethoprim, tetracycline, and erythromycin. This work provides useful information regarding contamination of RTE meat products with L. monocytogenes, which may have implications for food safety risks. [ABSTRACT FROM AUTHOR]

According to EU summary report on zoonoses, zoonotic agents and food‐borne outbreaks in 2017, Campylobacter was the most commonly reported gastrointestinal bacterial pathogen in humans in the EU. Unfortunately, the standard methods for the detection of thermotolerant Campylobacter spp. in foods are time‐consuming. Additionally, the qualified staff is obligatory. For this reason, new methods of pathogens detection are needed. The present work demonstrates that surface‐enhanced Raman scattering (SERS) is a reliable and fast method for detection of Campylobacter spp. in food samples. The proposed method combines the SERS measurements performed on an Ag/Si substrate with two initial steps of the ISO standard procedure. Finally, the principal component analysis (PCA) allows for statistical classification of the studied bacteria. By applying the proposed ISO‐SERS‐PCA method in the case of Campylobacter bacteria the total detection time may be reduced from 7 to 8 days required by ISO method to 3 to 4 days in the case of SERS‐based approach. [ABSTRACT FROM AUTHOR]

Abstract The sixty-seven nonpathogenic Listeria spp. strains isolated from food and food processing environments in Poland were examined for the presence of benzalkonium chloride (BC) resistance cassette (bcrABC) and four different variants of cadmium resistance determinants (cadA1-cadA4). All the strains were phenotypically resistant to cadmium and 22 among them were also resistant to BC. PCR-based analysis revealed that bcrABC cassette was harbored by 95.5% of the strains phenotypically resistant to BC. All of them harbored also either cadA1 or cadA2 genes (none carried cadA3 or cadA4), which corresponded to the presence of plasmids with two restriction patterns. The strains resistant to cadmium but susceptible to BC harbored only the cadA1 gene variant. DNA-DNA hybridization analysis showed that all the identified bcrABC , cadA 1 and cadA2 genes were located within plasmids, classified into 11 groups of RFLP profiles. Only one of the plasmids – pLIS1 of Listeria welshimeri (carrying bcrABC and cadA2) – was capable of efficient conjugal transfer from nonpathogenic Listeria isolates to a pathogenic Listeria monocytogenes strain. Analysis of the complete nucleotide sequence of pLIS1 (the first sequenced plasmid of L. welshimeri species) revealed the presence of genes involved in plasmid replication, stabilization and transfer as well as genes conferring resistance phenotypes. Comparative analysis showed that pLIS1 genome is highly similar to a group of plasmids originating from L. monocytogenes strains. A common feature of pLIS1 and its relatives, besides the presence of the resistance genes, is the presence of numerous transposable elements (TEs). The analysis revealed the important role of TEs in both promoting genetic rearrangements within Listeria spp. plasmids and the acquisition of resistance determinants. Highlights • Nonpathogenic cadmium resistant Listeria spp. carry either cadA1 or cadA2 genes • Majority of strains resistant to benzalkonium chloride carry bcrABC genes • The resistance determinants are located within plasmids of 11 RFLP patterns • Plasmid pLIS1 of L. welshimeri is capable of efficient conjugal transfer • pLIS1 contains bcrABC and cadA2 as well as numerous transposable elements [ABSTRACT FROM AUTHOR]

A total of 127 Listeria isolates from food and food processing environments, including 75 L. innocua , 49 L. welshimeri , 2 L. seeligeri and 1 L. grayi were tested for susceptibility to eight antimicrobials, benzalkonium chloride (BC), cadmium and arsenic. The isolates were also screened for the presence of extrachromosomal genetic elements - plasmids, and their restriction pattern types were determined. All strains were susceptible to ampicillin, ciprofloxacin, erythromycin, gentamicin, rifampicin, trimethoprim and vancomycin. Two of the L. innocua isolates showed resistance to tetracycline and minocycline. The resistance was determined by the presence of chromosomal localization of tet (M) gene, which was not integrated in the transposon Tn 916 -Tn 1545 family. Of analyzed isolates, 18.11% and 55.91% isolates were resistant to BC and cadmium, respectively, but all were susceptible to arsenic. Resistance to BC was correlated with resistance to cadmium - all BC resistant isolates were also resistant to cadmium. On the other hand, 67.61% of cadmium-resistant isolates were susceptible to BC, suggesting that cadmium and BC resistance were not always concurrent in Listeria species. 48.03% of isolates contained plasmids. The size of most of the identified replicons was in the range of 50–90 kb. All plasmids were classified into 12 groups with identical restriction pattern (I–XII). Interestingly, plasmids belonging to the same group were determined in isolates of the same species. Only in one case, plasmids with I-type profile were identified in L. innocua and L. welshimeri. There was an association between resistance to BC and plasmid DNA presence: all resistant isolates carried a plasmid. A correlation between resistance to cadmium and plasmid carriage was also observed in L. innocua and L. seeligeri isolates, but among resistant L. welshimeri, 23.08% of isolates did not have plasmids. This may suggest that resistance is associated with determinants located within the chromosome. To elucidate the adaptation strategies and ecology of Listeria spp., it is important to have a better understanding of its resistance to antimicrobials and environmental toxicants such as heavy metals and disinfectants. [ABSTRACT FROM AUTHOR]

The purpose of the present study was to determine the prevalence of thermophilic Campylobacter in poultry, pork, and beef meat at the retail level and to identify the main categories of meat representing the most significant reservoirs of Campylobacter. A monitoring study was conducted throughout Poland from 2009 to 2013. A total of 1,700 fresh meat samples were collected from supermarkets, large retail outlets, and smaller stores. Thermophilic Campylobacter species were detected in 690 (49.3%) of 1,400 poultry samples collected from retail trade. Strains were isolated from 50.2 and 41.1% of raw chicken and turkey meat samples, respectively, and from 50.1 and 42.6% of raw chicken and turkey giblets. The incidence of Campylobacter spp. on pork ( 10.6%) and beef ( 10.1 %) was significantly lower than on poultry. Campylobacter jejuni was the most prevalent Campylobacter species in chicken (46.6%), pork (68.6%), and beef (66.7%), and Campylobacter coli was the most frequently isolated Campylobacter species in turkey meat (71.2%). This study revealed that retail raw meats are often contaminated with Campylobacter; however, the prevalence of these pathogens is markedly different in different meats. Raw retail meats are potential vehicles for transmitting foodborne diseases, and our findings stress the need for increased implementation of hazard analysis critical control point programs and consumer food safety education efforts. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is a Gram-positive bacterium causing rare but dangerous cases of disease in humans and animals. The β-lactams penicillin G and ampicillin are the antibiotics of choice in the treatment of listeriosis. Recently, lmo1941, encoding a surface protein of L. monocytogenes with unknown function, was identified as a gene transcriptionally upregulated under penicillin G pressure. In this study, the effect of lmo1941 knockout on the susceptibility of L. monocytogenes to β-lactams was examined. Deletion mutant in lmo1941 was constructed and subjected to studies, which revealed that the deletion of lmo1941 had no effect on susceptibility and tolerance to penicillin G and ampicillin but resulted, however, in increased susceptibility of L. monocytogenes to several cephalosporins. Subsequently, the potential effect of lmo1941 mutation on the cell wall of L. monocytogenes was investigated. The analysis revealed quantitative changes in the muropeptide profile of peptidoglycan and a decrease in density of the high-density zone of cell wall of the mutant strain. Both these changes were observed in cells taken from the stationary phase. These results indicate that the surface protein Lmo1941 affects peptidoglycan composition and cell wall structure of L. monocytogenes in the stationary phase of growth. [ABSTRACT FROM AUTHOR]

Abstract: A total of 471 Listeria monocytogenes isolates from different types of food and food-related sources in Poland during 2004–2010 were examined. This number includes 200 isolates from fish, 144 from fresh and frozen vegetables, 43 ready-to-eat products (deli foods, cold cuts), 13 from dairy products, 16 from raw meats, 15 from confectionery products and 40 directly from processing plants. All isolates were subjected to serotyping and lineage assays using PCR, and antimicrobial susceptibility using E-test and a broth microdilution method. Of all isolates, 256 (54.4%), 120 (25.5%), 59 (12.5%), 36 (7.6%) were identified as serotypes 1/2a (or 3a), 1/2c (or 3c), 1/2b (or 3b or 7), and 4b (or 4d or 4e), respectively. A direct correlation between the most common serotypes and three L. monocytogenes lineages was also observed. All L. monocytogenes isolates belonged to lineages I (20.2%) and II (79.8%). All strains were sensitive to ampicillin, amoxicillin, gentamicin, erythromycin, trimethoprim, rifampicin, vancomycin, chloramphenicol and sulfamethoxazol. Two of the L. monocytogenes strains (0.42%) showed phenotypic resistance. One strain was resistant to tetracycline and minocycline due to the presence of tet(M). It did not carry gene int, which may indicate that the tet(M) gene in this strain was not integrated in the transposon Tn916-Tn1545 family. The resistance of the second strain to ciprofloxacin and norfloxacin was attributed to active efflux associated with overexpression of gene lde. Our data indicate the low prevalence of antimicrobial resistance among L. monocytogenes isolates from food and food-related sources in Poland. [Copyright &y& Elsevier]

Abstract: This study presents the results of investigations on the susceptibility of Campylobacter spp. strains isolated from chicken meat and giblets to fluorochinolones (ciprofloxacin), macrolides (erythromycin), tetracyclines (tetracycline) and aminoglycosides (gentamicin) andV an analysis of the molecular mechanisms of resistance to the selected antibiotics. Between January 2008 and December 2009 a total of 218 samples of chicken meat and giblets from retail trade in Poland were examined. Campylobacter bacteria were found in 143 samples, that is in 65.6% of the total number embraced by the study. Campylobacter coli was the most ubiquitous – its presence was determined in 108 samples out of 143 (75.5%), whereas Campylobacter jejuni was found in 35 of the contaminated samples (24.5%). The results obtained point to the high percentage (97.9%) of Campylobacter isolates resistant to ciprofloxacin. 92 strains (64.3%) were resistant to tetracycline, 14 (9.1%) to erythromycin and only 9 (6.3%) to gentamicin. Moreover, ten out of the 143 resistant Campylobacter strains (7.0%) were found to be resistant to at least three unrelated antibiotics. [Copyright &y& Elsevier]

In 2007 and 2008, a monitoring study was carried out in Poland to examine the occurrence of thermotolerant Campylobacter spp. in raw and cooked chicken products available on the retail market. A total of 912 samples were tested: 443 samples of raw chicken meat, 146 samples of giblets, and 323 ready-to-eat poultry products (150 samples of spit-roasted chicken, 56 samples of smoked chicken, and 117 samples of pâté and cold meats). A high level of contamination of raw chicken meat (51.7% o samples) and chicken giblets (47.3% of samples) was detected. However, thermotolerant Campylobacter spp. were found in only 1.2% of the ready-to-eat poultry products. [ABSTRACT FROM AUTHOR]

Background: Bacterial penicillin-binding proteins (PBPs) can be visualized by their ability to bind radiolabeled or fluorescent β-lactam derivatives both whole cells and membrane/cell enriched fractions. Analysis of the Listeria monocytogenes genome sequence predicted ten genes coding for putative PBPs, but not all of their products have been detected in studies using radiolabeled antibiotics, thus hindering their characterization. Here we report the positive identification of the full set of L. monocytogenes PBPs and the characteristics of the hitherto undescribed PBPD2 (Lmo2812). Results: Eight L. monocytogenes PBPs were identified by the binding of fluorescent β-lactam antibiotic derivatives Boc-FL, Boc-650 and Amp-Alexa430 to proteins in whole cells or membrane/cell wall extracts. The gene encoding a ninth PBP (Lmo2812) was cloned and expressed in Escherichia coli as a His-tagged protein. The affinity purified recombinant protein had DD-carboxypeptidase activity and preferentially degraded low-molecular-weight substrates. L. monocytogenes mutants lacking the functional Lmo2812 enzyme were constructed and, compared to the wild-type, the cells were longer and slightly curved with bent ends. Protein Lmo1855, previously designated PBPD3, did not bind any of the antibiotic derivatives tested, similarly to the homologous enterococcal protein VanY. Conclusions: Nine out of the ten putative L. monocytogenes PBP genes were shown to encode proteins that bind derivatives of β-lactam antibiotics, thus enabling their positive identification. PBPD2 (Lmo2812) was not visualized in whole cell extracts, most probably due to its low abundance, but it was shown to bind Boc-FL after recombinant overexpression and purification. Mutants lacking Lmo2812 and another low molecular mass (LMM) PBP, PBP5 (PBPD1) - both with DD-carboxypeptidase activity - displayed only slight morphological alterations, demonstrating that they are dispensable for cell survival and probably participate in the latter stages of peptidoglycan synthesis. Since Lmo2812 preferentially degrades low-molecular- mass substrates, this may indicate a role in cell wall turnover. [ABSTRACT FROM AUTHOR]

The study describes the characterization of Listeria monocytogenes isolated from the general 2017–2019 national official control and monitoring sampling program. A total of 60,928 of ready-to-eat (RTE) food products were collected in retail in Poland, while the number of L. monocytogenes contaminated samples was 67 (0.1%). The majority of the strains belonged to molecular serotype IVb followed by IIa, frequently associated with human listeriosis. Furthermore, 61.2% of the isolates were resistant at least to one of the tested antimicrobials: penicillin, ampicillin, meropenem, erythromycin, sulfamethoxazole-trimethoprim, amoxicillin-clavulanic acid, ciprofloxacin, chloramphenicol, gentamicin, vancomycin, tetracycline and rifampicin. Virulence genes inlA , inlC , inlJ and lmo2672 were detected in all of the isolates. In our study the llsX gene (encoding LLS) exhibited 11.6% positivity. The 32 strains were grouped into 12 clonal complexes (CCs) which belong to the major clones that are in circulation in Europe. Among them, seven strains with the cgMLST close relatedness (CC2) were isolated from diverse food sectors, underlining a large circulation of this clone in Poland, most likely from multiple introduction sources. Additionally, two RTE strains CC6 and one CC37 were identified as closely related by cgMLST to two publicly available genomes of clinical strains isolated in Poland in 2012–2013. These results indicate the large strain circulation and point to RTE food products as a potential source of human listeriosis. The present study provided data to capture the contamination status of L. monocytogenes in foods at the retail level in Poland and assess the potential risk of this pathogen for human safety. • L. monocytogenes was isolated from 0.1% of the RTE food samples. • The most frequently identified molecular serogroups were IVb (51%) and IIa (42%). • 48% strains belonged to the major clonal complexes (CCs) circulating in Europe. • Seven strains (CC2) close relatedness were isolated from diverse food sectors. • Two strains CC6 and one C37 were closely related to clinical strains in Poland. [ABSTRACT FROM AUTHOR]

Campylobacter-associated gastroenteritis remains an important cause of morbidity worldwide, and some evidence suggests that poultry is an important source of this foodborne infection in humans. This study was conducted to analyze the prevalence and genetic background of resistance of 149 Campylobacterjejuni and 54 Campylobacter coli strains isolated from broiler chicken carcasses and from stool samples of infected children in Poland from 2003 through 2005. Nearly all isolates were susceptible to macrolides and aminoglycosides. The highest resistance in both human and chicken strains was observed for ciprofloxacin (more than 40%), followed by ampicillin (13 to 21%), and tetracycline (8 to 29%). Resistance to ampicillin and tetracycline rose significantly between 2003 and 2005. Slight differences in resistance between human and chicken isolates indicate that although chicken meat is not the only source of Campylobacter infection in our population, it can be involved in the transmission of drug-resistant Campylobacter strains to humans. [ABSTRACT FROM AUTHOR]

Abstract: We report on the cloning of the structural gene for penicillin-binding protein 5 (PBP5), lmo2754. We also describe the enzymatic activity of PBP5 and characterize a mutant lacking this activity. Purified PBP5 has dd-carboxypeptidase activity, removing the terminal d-alanine residue from murein pentapeptide side chains. It shows higher activity against low molecular weight monomeric pentapeptide substrates compared to dimeric pentapeptide compound. Similarly, PBP5 preferentially cleaves monomeric pentapeptides present in high-molecular weight murein sacculi. A Listeria monocytogenes mutant lacking functional PBP5 was constructed. Cells of the mutant are viable, showing that the protein is dispensable for growth, but grow slower and have thickened cell walls. [Copyright &y& Elsevier]

Bacteria of the genus Listeria (phylum Firmicutes) include both human and animal pathogens, as well as saprophytic strains. A common component of Listeria spp. genomes are plasmids, i.e., extrachromosomal replicons that contribute to gene flux in bacteria. This study provides an in-depth insight into the structure, diversity and evolution of plasmids occurring in Listeria strains inhabiting various environments under different anthropogenic pressures. Apart from the components of the conserved plasmid backbone (providing replication, stable maintenance and conjugational transfer functions), these replicons contain numerous adaptive genes possibly involved in: (i) resistance to antibiotics, heavy metals, metalloids and sanitizers, and (ii) responses to heat, oxidative, acid and high salinity stressors. Their genomes are also enriched by numerous transposable elements, which have influenced the plasmid architecture. The plasmidome of Listeria is dominated by a group of related replicons encoding the RepA replication initiation protein. Detailed comparative analyses provide valuable data on the level of conservation of these replicons and their role in shaping the structure of the Listeria pangenome, as well as their relationship to plasmids of other genera of Firmicutes, which demonstrates the range and direction of flow of genetic information in this important group of bacteria. [ABSTRACT FROM AUTHOR]

Phenotypic and genotypic resistance to benzalkonium chloride (BC), cadmium and arsenic was tested (by susceptibility assays and molecular methods) in 287 Listeria monocytogenes strains isolated from fish and fish products, and food-producing factories in Poland. Overall, 40% of the isolates were resistant to BC, 56% to cadmium and 41% to arsenic (57% displayed resistance to more than one of the tested compounds). Among BC-resistant isolates, the most commonly detected resistance determinant was the qacH gene (83%). Three distinct types of cadA gene determining resistance to cadmium were detected, with the cadA1 variant predominant (88%), while most arsenic-resistant isolates (86%) harbored the arsA gene associated with a Tn 554- like transposon (one strain harbored two copies of arsA in different arsenic resistance cassettes). 53% of all tested isolates contained plasmids (from 4 kb to > 90 kb in size), which were classified into 11 groups (p1–p11) based on their restriction patterns. Interestingly, 12 isolates harbored the small mobilizable pLMST6-like plasmid pLIS3 encoding multidrug efflux pump EmrC. Clustering analysis of PFGE patterns revealed that these isolates represent several diverse bacterial populations, which strongly suggests mobility of the pLMST6-like plasmids among L. monocytogenes strains and their role in dissemination of BC resistance. • Among 287 Listeria monocytogenes isolates, 40% were resistant to benzalkonium chloride, 56% to cadmium and 41% to arsenic. • 83% of isolates resistant to benzalkonium chloride carried the qacH gene. • 88% of cadmium-resistant isolates carried the cadA1 gene. • 86% of isolates resistant to arsenic harbored the Tn 554 associated arsA gene. • 4% of isolates carried a mobilizable pLMST6-like plasmid pLIS3 encoding multidrug efflux pump EmrC. [ABSTRACT FROM AUTHOR]

The aim of the present study is to investigate the prevalence and genetic diversity of Listeria monocytogenes in various fresh and frozen vegetable products available in Poland. The samples were collected at retail market within the framework of national official control and monitoring program. In the years 2016–2019 a total of 49 samples out of 8712 collected vegetable samples were positive for L. monocytogenes. Our findings demonstrated that the occurrence of L. monocytogenes in various vegetable products was generally low, on average only 0.56% in the studied years. All isolates were susceptible to 11 antimicrobial agents: penicillin, ampicillin, meropenem, erythromycin, sulfamethoxazole-trimethoprim, amoxicillin-clavulanic acid, ciprofloxacin, chloramphenicol, gentamicin, vancomycin, and tetracycline. All of them harbored virulence-associated genes (inlA , inlC , and lmo2672), 82% harbored inlJ gene and few of them (22%) also possessed the llsX gene. The majority of collected isolates (65%) belonged to molecular serogroup 1/2a-3a, followed by 4ab-4b-4d-4e (33%), and only one to serogroup 1/2b-3b-7 (2%). Isolates yielded 18 different restriction profiles, revealing a large cluster of contamination linked to frozen corn (21 strains) and distributed in 3 pulsotypes. MLST analysis classified selected isolates into nine clonal complexes (CCs). The obtained results contribute to characterizing the diversity of L. monocytogenes isolated from various vegetable products in Poland and their impact on food safety and public health. • 0.56% vegetable products in Poland were positive for L. monocytogenes. • The most prevalent was serogroup 1/2a-3a (65%). • All isolates were susceptible to tested antimicrobials. • The inlA , inlC and lmo2672 genes were observed in all isolates. • MLST analysis classified selected isolates into nine clonal complexes. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is a pathogen responsible for severe cases of food poisoning. Listeria spp. strains occurring in soil and water environments may serve as a reservoir of resistance determinants for pathogenic L. monocytogenes strains. A large collection of Listeria spp. strains (155) isolated from natural, agricultural, and urban areas was screened for resistance to heavy metals and metalloids, and the presence of resistance determinants and extrachromosomal replicons. Of the tested strains, 35% were resistant to cadmium and 17% to arsenic. Sequence analysis of resistance plasmids isolated from strains of Listeria seeligeri and Listeria ivanovii, and the chromosome of L. seeligeri strain Sr73, identified a novel variant of the cadAC cadmium resistance efflux system, cadA6, that was functional in L. monocytogenes cells. The cadA6 cassette was detected in four Listeria species, including strains of L. monocytogenes, isolated from various countries and sources—environmental, food-associated, and clinical samples. This resistance cassette is harbored by four novel composite or non-composite transposons, which increases its potential for horizontal transmission. Since some cadAC cassettes may influence virulence and biofilm formation, it is important to monitor their presence in Listeria spp. strains inhabiting different environments. [ABSTRACT FROM AUTHOR]

The aim of the study was to characterize strains of Listeria monocytogenes isolated from ready to eat (RTE) products collected as part of official food control and monitoring in Poland. A total of 105 L. monocytogenes isolates from RTE products: 54- cakes and 51 – delicatessen products were examined. The presence L. monocytogenes in cakes and delicatessen products was 0.4% and 0.7% respectively suggesting the level of contamination of RTE products with L. monocytogenes is very low. [ABSTRACT FROM AUTHOR]

Bacillus cereus sensu lato can contaminate food and cause food poisoning by producing toxins such as cereulide, toxin BL, and cytotoxin K. In this study, we retrospectively analyzed B. cereus sensu lato from retail food products and food poisoning cases using PCR methods to determine their virulence profiles. A new toxin profile, encoding all four toxins ( hbl , nhe , cytK , ces ), was found in 0.4% of isolates. The toxin profiles, classified into A-J, revealed that 91.8% harbored nhe genes, while hbl , cytK , and ces were detected in 43.8%, 46.9%, and 4.2% of isolates, respectively. Whole-genome sequencing (WGS) identified four distinct species within the B. cereus group, with 21 isolates closely related to B. cereus sensu stricte , 25 to B. mosaicus , 2 to B. toyonensis , and 1 to B. mycoides . Three novel sequence types (STs 3297, 3298, 3299) were discovered. Antibiotic resistance genes were common, with 100% of isolates carrying beta-lactam resistance genes. Fosfomycin (80%), vancomycin (8%), streptothricin (6%), tetracycline (4%), and macrolide resistance (2%) genes were also detected. These results highlight the genetic diversity and antibiotic resistance potential of B. cereus sensu lato strains in Polish food products.

The prevalence, count and molecular identification of Campylobacter spp. in Polish poultry meat were analysed. 181 samples of meat from chicken (70), turkey (47), duck (54) and goose (10) were studied. Campylobacter spp. was found in 64% of meat samples. The highest prevalence of this pathogen was detected for duck meat. On average 80% of duck samples were contaminated with Campylobacter spp. The counts of Campylobacter spp. in positive samples remained under ten colony forming units per gram of product in 59% of poultry meat. C. jejuni was more frequently detected in poultry meat than C. coli.

In this study, we investigated the molecular mechanisms involved in erythromycin resistance in the first resistant Campylobacter strains isolated from chicken meat in Poland, and analyzed their genetic relatedness. A total of 297 samples of raw chicken meat and giblets from retail trade in the Warsaw area collected between 2006 and 2009 were examined. Among 211 Campylobacter strains (52 C. jejuni and 159 C. coli), 10 C. coli isolates (4.7%) were resistant to erythromycin. All the C. jejuni strains were susceptible. Among the high-level macrolide-resistant isolates, two different point mutations within the domain V of the 23S rRNA gene were observed. Eight of the strains had adenine→guanine transitions at position 2075, two other isolates at position 2074. Sequence analysis of ribosomal proteins L4 (rplD) and L22 (rplV) indicated that ribosomal protein modifications did not contribute to macrolide resistance. A mutation in the inverted repeat in the cmeR and cmeABC intergenic region was found in a single resistant strain. The genetic relatedness of Campylobacter isolates showed that two resistant strains obtained from the same production plant in a 2-month interval were genetically identical. The risk of transmission of resistant strains via the food chain highlights the need for constant monitoring of resistance in Campylobacter isolates of human and animal hosts.

Campylobacter is recognized as a major cause of human acute bacterial enteritis. The incidence of human Campylobacter infection has increased markedly in both developed and developing countries and, more significantly, so has rapid emergence of antibiotic-resistant Campylobacter strains. It is caused by improper applying antibiotics in treating people and too frequent applying these substances in the animal husbandry. In this review, the patterns of emerging resistance to the antimicrobial agents useful in treatment of the disease are presented and the mechanisms of resistance to these drugs in Campylobacter spp. are discussed.

Microbial contamination of a water distribution system was examined. The number and the taxonomy of non-pigmented and pigmented heterotrophic bacteria (HB), number of bacteria (Pseudomonas sp., Enterococcus sp., Campylobacter sp., Yersinia sp., representatives of the Enterobacteriaceae, coagulase-positive staphylococci, and C. pefringens) in the bulk water phase, biomass of zoogloeal aggregates of bacteria, fungi, algae, protozoa and rotifers (ZABFAPR) (separated from the above on 5 microm pore size filters) and in pipe sediments was determined. An increased number of HB occurred at the sampling sites situated as close as 4.2 km to the Water Treatment Plant (WTP), and was especially significant at 10.3 km. It was shown that the main reservoir of hygienically relevant bacteria did not occur in the water phase which is monitored in routine control analyses carried out by the WTP laboratories, but in the ZABFAPR biomass not monitored so far.

The antibiotic susceptibilities and capabilities to induce beta-lactamases were studied in multiple Escherichia coli murein (peptidoglycan) hydrolase mutants. E. coli mutants lacking either three amidases, three amidases and one lytic transglycosylase, or six lytic transglycosylases showed higher levels of susceptibility to bacitracin, erythromycin, gallidermin, and vancomycin than the wild type. Mutant cells without three amidases lost viability in the presence of vancomycin and gallidermin, whereas the wild type was resistant to both antibiotics. Beta-lactamase induction was studied after introduction of a plasmid carrying the ampC and ampR genes. Upon addition of cefoxitin to the growth medium, the wild type as well as a mutant lacking all known amidases and DD-endopeptidases induced beta-lactamase, whereas a mutant lacking all known lytic transglycosylases was unable to induce beta-lactamase, showing that lytic transglycosylase activity is essential for beta-lactamase induction. Consequently, cells lacking lytic transglycosylase activity lysed in the presence of penicillin, despite the presence of the inducible beta-lactamase system. We discuss the potential of murein hydrolase inhibitors for antibiotic therapy.

Cells of a mutant of Listeria monocytogenes lacking functional PBP5, an enzyme with DD-carboxypeptidase activity, make thicker cells walls. In this study we show that the muropeptide profile of the mutant, obtained after HPLC analysis of a muramidase digest of cell wall murein, differs from that for the wild type strain. The main differences embrace strongly reduced disaccharide-tripeptide content, strongly increased amounts of pentapeptide-containing muropeptides and a shift in profile from less cross-linked muropeptides (monomers, dimers) towards more highly cross-linked ones.

Previously found DNA sequence (iam), correlated with the invasiveness of Campylobacter strains, became a starting point for present investigation. Main goal of this study was to isolate number of Campylobacter strains from chicken carcasses, to determine their taxonomic position and to establish the presence of iam sequence in their genoms. It was found, that invasion associated marker is present in all Campylobacter coli strains and in majority but not all Campylobacter jejuni. This may confirm the idea that the marker is not only responsible for diarrhea in humans but also may be important in the colonization of chicken guts.

Intact Listeria monocytogenes cells or membranes isolated from them were treated with [3H]penicillin to allow identification of the penicillin binding proteins (PBPs) located in the cytoplasmic membrane. In the former case the PBPs were released from the cells following disruption of the cell wall murein with Listeria monocytogenes bacteriophage lysin. The procedure described by Dougherty et al. (1996) for Escherichia coli, with some modifications, was used to evaluate the M(r)s of the individual PBPs and allowed direct quantitation of their copy number.