The influence of stress factors on selected phenotypic and genotypic features of Listeria monocytogenes - a pilot study.

BACKGROUND: Listeria monocytogenes are Gram-positive rods, widespread in the environment due to their wide tolerance to changing conditions. The apilot study aimed to assess the impact of six various stresses (heat, cold, osmotic, acid, alkali, frozen) on phenotypic features: MIC of antibiotics (penicillin, ampicillin, meropenem, erythromycin, co-trimoxazole; gradient stripes), motility, ability to form a biofilm (crystal violet method) and growth rate (OD and quantitative method), expression level of sigB (stress induced regulator of genes), agrA, agrB (associated with biofilm formation) and lmo2230, lmo0596 (acid and alkali stress) (qPCR) for three strains of L. monocytogenes. RESULTS: Applied stress conditions contributed to changes in phenotypic features and expression levels of sigB, agrA, agrB, lmo2230 and lmo0596. Stress exposure increased MIC value for penicillin (ATCC 19111 - alkaline stress), ampicillin (472CC - osmotic, acid, alkaline stress), meropenem (strains: 55 C - acid, alkaline, o smotic, frozen stress; 472CC - acid, alkaline stress), erythromycin (strains: 55 C - acid stress; 472CC - acid, alkaline, osmotic stress; ATCC 19111 - osmotic, acid, alkaline, frozen stress), co-trimoxazole (strains: 55 C - acid stress; ATCC 19111 - osmotic, acid, alkaline stress). These changes, however, did not affect antibiotic susceptibility. The strain 472CC (a moderate biofilm former) increased biofilm production after exposure to all stress factors except heat and acid. The ATCC 19111 (a weak producer) formed moderate biofilm under all studied conditions except cold and frozen stress, respectively. The strain 55 C became a strong biofilm producer after exposure to cold and produced a weak biofilm in response to frozen stress. Three tested strains had lower growth rate (compared to the no stress variant) after exposure to heat stress. It has been found that the sigB transcript level increased under alkaline (472CC) stress and the agrB expression increased under cold, osmotic (55 C, 472CC), alkali and frozen (472CC) stress. In contrast, sigB transcript level decreased in response to acid and frozen stress (55 C), lmo2230 transcript level after exposure to acid and alkali stress (ATCC 19111), and lmo0596 transcript level after exposure to acid stress (ATCC 19111). CONCLUSIONS: Environmental stress changes the ability to form a biofilm and the MIC values of antibiotics and affect the level of expression of selected genes, which may increase the survival and virulence of L. monocytogenes. Further research on a large L. monocytogenes population is needed to assess the molecular mechanism responsible for the correlation of antibiotic resistance, biofilm formation and resistance to stress factors.

Genomic and pathogenicity islands of Listeria monocytogenes-overview of selected aspects.

Listeria monocytogenes causes listeriosis, a disease characterized by a high mortality rate (up to 30%). Since the pathogen is highly tolerant to changing conditions (high and low temperature, wide pH range, low availability of nutrients), it is widespread in the environment, e.g., water, soil, or food. L. monocytogenes possess a number of genes that determine its high virulence potential, i.e., genes involved in the intracellular cycle (e.g., prfA, hly, plcA, plcB, inlA, inlB), response to stress conditions (e.g., sigB, gadA, caspD, clpB, lmo1138), biofilm formation (e.g., agr, luxS), or resistance to disinfectants (e.g., emrELm, bcrABC, mdrL). Some genes are organized into genomic and pathogenicity islands. The islands LIPI-1 and LIPI-3 contain genes related to the infectious life cycle and survival in the food processing environment, while LGI-1 and LGI-2 potentially ensure survival and durability in the production environment. Researchers constantly have been searching for new genes determining the virulence of L. monocytogenes. Understanding the virulence potential of L. monocytogenes is an important element of public health protection, as highly pathogenic strains may be associated with outbreaks and the severity of listeriosis. This review summarizes the selected aspects of L. monocytogenes genomic and pathogenicity islands, and the importance of whole genome sequencing for epidemiological purposes.

Assessment of the influence of selected stress factors on the growth and survival of Listeria monocytogenes.

BACKGROUND: Listeria monocytogenes are Gram-positive rods, which are the etiological factor of listeriosis. L. monocytogenes quickly adapts to changing environmental conditions. Since the main source of rods is food, its elimination from the production line is a priority. The study aimed to evaluate the influence of selected stress factors on the growth and survival of L. monocytogenes strains isolated from food products and clinical material. RESULTS: We distinguished fifty genetically different strains of L. monocytogenes (PFGE method). Sixty-two percent of the tested strains represented 1/2a-3a serogroup. Sixty percent of the rods possessed ten examined virulence genes (fbpA, plcA, hlyA, plcB, inlB, actA, iap, inlA, mpl, prfA). Listeria Pathogenicity Island 1 (LIPI-1) was demonstrated among 38 (76.0%) strains. Majority (92.0%) of strains (46) were sensitive to all examined antibiotics. The most effective concentration of bacteriophage (inhibiting the growth of 22 strains; 44.0%) was 5 × 108 PFU. In turn, the concentration of 8% of NaCl was enough to inhibit the growth of 31 strains (62.0%). The clinical strain tolerated the broadest pH range (3 to 10). Five strains survived the 60-min exposure to 70˚C, whereas all were alive at each time stage of the cold stress experiment. During the stress of cyclic freezing-defrosting, an increase in the number of bacteria was shown after the first cycle, and a decrease was only observed after cycle 3. The least sensitive to low nutrients content were strains isolated from frozen food. The high BHI concentration promoted the growth of all groups. CONCLUSIONS: Data on survival in stress conditions can form the basis for one of the hypotheses explaining the formation of persistent strains. Such studies are also helpful for planning appropriate hygiene strategies within the food industry.

Antibiotic Resistance of Enterococcus spp. Isolated from the Urine of Patients Hospitalized in the University Hospital in North-Central Poland, 2016-2021.

Urinary Tract Infections (UTIs) are common outpatient and inpatient infections, often treated with empirical therapy. Enterococcus spp. is responsible for about 10% of UTIs. This study aimed to determine the necessity of changing the empirical treatment of UTIs caused by Enterococcus spp. The evaluation was performed for 542 Enterococcus strains isolated from urine samples in the years 2016-2021. We identified three Enterococcus species that were found: E. faecalis (389, 71.8%), E. faecium (151, 27.8%) and E. gallinarum (2, 0.4%). E. faecalis was the dominant species every year. Among E. faecalis, the most prevalent was resistance to norfloxacin (51.4%). Almost all E. faecium strains (150, 99.3%) were resistant to beta-lactams and norfloxacin. Eighty-three strains (55.0%) were resistant to vancomycin and 72 (47.7%) to teicoplanin. E. faecium strains showed a significantly higher percentage of resistance mechanisms GRE (Glicopeptide-Resistant Enterococcus) (72, 48.7%) and VRE (Vancomycin-Resistant Enterococcus) (11, 7.3%), while only five strains of E. feacalis showed a VRE mechanism (1.3%). In the therapy of E. faecalis UTIs, ampicillin and imipenem still remain effective. However, the above-mentioned antibiotics, as well as fluoroquinolones, are not recommended in the treatment of UTIs of E. faecium etiology.

Effect of Radiant Catalytic Ionization and Ozonation on Salmonella spp. on Eggshells.

Three Salmonella enterica strains were used in the study (serovars: S. enteritidis, S. typhimurim and S. virchow). This study evaluated the efficacy of radiant catalytic ionization (RCI) and ozonation against Salmonella spp. on eggshell (expressed as log CFU/egg). The egg surface was contaminated three different bacterial suspension (103 CFU/mL, 105 CFU/mL and 108 CFU/mL) with or without poultry manure. Experiments were conducted at 4 °C and 20 °C in three different time period: 30 min, 60 min and 120 min. Treatment with RCI reduced Salmonella numbers from 0.26 log CFU/egg in bacterial suspension 108 CFU/mL, 4 °C and 20 °C, with manure for 30 min to level decrease in bacteria number below the detection limit (BDL) in bacterial suspension 105 CFU/mL, 20 °C, with or without manure for 120 min. The populations of Salmonella spp. on eggs treated by ozonizer ranged from 0.20 log CFU/egg in bacteria suspension 108 CFU/mL, 20 °C, with manure for 30 min to 2.73 log CFU/egg in bacterial suspension 105 CFU/mL, 20 °C, with manure for 120 min. In all treatment conditions contamination with poultry manure decrease effectiveness the RCI and ozonation. In summary, RCI technology shows similar effectiveness to the ozonation, but it is safer for poultry plant workers and consumers.

The Role of Psychobiotics in Supporting the Treatment of Disturbances in the Functioning of the Nervous System-A Systematic Review.

Stress and anxiety are common phenomena that contribute to many nervous system dysfunctions. More and more research has been focusing on the importance of the gut-brain axis in the course and treatment of many diseases, including nervous system disorders. This review aims to present current knowledge on the influence of psychobiotics on the gut-brain axis based on selected diseases, i.e., Alzheimer's disease, Parkinson's disease, depression, and autism spectrum disorders. Analyses of the available research results have shown that selected probiotic bacteria affect the gut-brain axis in healthy people and people with selected diseases. Furthermore, supplementation with probiotic bacteria can decrease depressive symptoms. There is no doubt that proper supplementation improves the well-being of patients. Therefore, it can be concluded that the intestinal microbiota play a relevant role in disorders of the nervous system. The microbiota-gut-brain axis may represent a new target in the prevention and treatment of neuropsychiatric disorders. However, this topic needs more research. Such research could help find effective treatments via the modulation of the intestinal microbiome.

The effect of fluoroquinolones and antioxidans on biofilm formation by Proteus mirabilis strains.

BACKGROUND: Fluoroquinolones are a group of antibiotics used in urinary tract infections. Unfortunately, resistance to this group of drugs is currently growing. The combined action of fluoroquinolones and other antibacterial and anti-biofilm substances may extend the use of this therapeutic option by clinicians. The aim of the study was to determine the effect of selected fluoroquinolones and therapeutic concentrations of ascorbic acid and rutoside on biofilm formation by Proteus mirabilis. MATERIALS AND METHODS: The study included 15 strains of P. mirabilis isolated from urinary tract infections in patients of the University Hospital No. 1 dr A. Jurasz in Bydgoszcz (Poland). The metabolic activity of the biofilm treated with 0.4 mg/ml ascorbic acid, 0.02 µg/ml rutoside and chemotherapeutic agents (ciprofloxacin, norfloxacin) in the concentration range of 0.125-4.0 MIC (minimum inhibitory concentration) was assessed spectrophotometrically. RESULTS: Both ciprofloxacin and norfloxacin inhibited biofilm formation by the tested strains. The biofilm reduction rate was correlated with the increasing concentration of antibiotic used. No synergism in fluoroquinolones with ascorbic acid, rutoside or both was found. The ascorbic acid and rutoside combination, however, significantly decreased biofilm production. CONCLUSIONS: Our research proves a beneficial impact of ascorbic acid with rutoside supplementation on biofilm of P. mirabilis strains causing urinary tract infections.

Comparison of Selected Phenotypic Features of Persistent and Sporadic Strains of Listeria monocytogenes Sampled from Fish Processing Plants.

(1) Background: The main source of transmission of Listeria monocytogenes is contaminated food, e.g., fish and meat products and raw fruit and vegetables. The bacteria can remain for 13 years on machines in food processing plants, including fish plants. (2) Methods: A total of 720 swabs were collected from a salmon filleting line. The research material consisted of 62 (8.6%) L. monocytogenes isolates. Pulsed Field Gel Electrophoresis (PFGE) allowed detecting a pool of persistent strains. All persistent strains (n = 6) and a parallel group of strains collected sporadically (n = 6) were characterized by their ability to invade HT-29 cells, biofilm formation ability, and minimum bactericidal concentrations (MBC) of selected disinfectants. (3) Results: Among the obtained isolates, 38 genetically different strains were found, including 6 (15.8%) persistent strains. The serogroup 1/2a-3a represented 28 strains (73.7%), including the persistent ones. There were no significant differences in invasiveness between the persistent and sporadic strains. The persistent strains tolerated higher concentrations of the tested disinfectants, except for iodine-based compounds. The persistent strains initiated the biofilm formation process faster and formed it more intensively. (4) Conclusions: The presence of persistent strains in the food processing environment is a great challenge for producers to ensure consumer safety. This study attempts to elucidate the phenotypic characteristics of persistent L. monocytogenes strains.

Staphylococcal Enterotoxin Genes in Coagulase-Negative Staphylococci-Stability, Expression, and Genomic Context.

In the current study, we screened a collection of coagulase-negative staphylococci (CoNS) isolates for orthologues of staphylococcal enterotoxins (SEs) involved in S. aureus-related staphylococcal food poisoning (SFP). The amplicons corresponding to SEs were detected in S. chromogenes, S. epidermidis, S. haemolyticus, S. borealis, S. pasteuri, S. saprophyticus, S. vitulinus, S. warneri, and S. xylosus. All amplicons were sequenced and identified as parts of known S. aureus or S. epidermidis SE genes. Quantitative real-time PCR allowed determining the relative copy number of each SE amplicon. A significant portion of the amplicons of the sea, seb, sec, and seh genes occurred at low copy numbers. Only the amplicons of the sec gene identified in three isolates of S. epidermidis displayed relative copy numbers comparable to sec in the reference enterotoxigenic S. aureus and S. epidermidis strains. Consecutive passages in microbiological media of selected CoNS isolates carrying low copy numbers of sea, seb, sec, and seh genes resulted in a decrease of gene copy number. S. epidermidis isolates harbored a high copy number of sec, which remained stable over the passages. We demonstrated that enterotoxin genes may occur at highly variable copy numbers in CoNS. However, we could identify enterotoxin genes only in whole-genome sequences of CoNS carrying them in a stable form at high copy numbers. Only those enterotoxins were expressed at the protein level. Our results indicate that PCR-based detection of enterotoxin genes in CoNS should always require an additional control, like analysis of their presence in the bacterial genome. We also demonstrate S. epidermidis as a CoNS species harboring SE genes in a stable form at a specific chromosome site and expressing them as a protein.

Two Faces of Fermented Foods-The Benefits and Threats of Its Consumption.

In underdeveloped and developing countries, due to poverty, fermentation is one of the most widely used preservation methods. It not only allows extending the shelf life of food, but also brings other benefits, including inhibiting the growth of pathogenic microorganisms, improving the organoleptic properties and product digestibility, and can be a valuable source of functional microorganisms. Today, there is a great interest in functional strains, which, in addition to typical probiotic strains, can participate in the treatment of numerous diseases, disorders of the digestive system, but also mental diseases, or stimulate our immune system. Hence, fermented foods and beverages are not only a part of the traditional diet, e.g., in Africa but also play a role in the nutrition of people around the world. The fermentation process for some products occurs spontaneously, without the use of well-defined starter cultures, under poorly controlled or uncontrolled conditions. Therefore, while this affordable technology has many advantages, it can also pose a potential health risk. The use of poor-quality ingredients, inadequate hygiene conditions in the manufacturing processes, the lack of standards for safety and hygiene controls lead to the failure food safety systems implementation, especially in low- and middle-income countries or for small-scale products (at household level, in villages and scale cottage industries). This can result in the presence of pathogenic microorganisms or their toxins in the food contributing to cases of illness or even outbreaks. Also, improper processing and storage, as by well as the conditions of sale affect the food safety. Foodborne diseases through the consumption of traditional fermented foods are not reported frequently, but this may be related, among other things, to a low percentage of people entering healthcare care or weaknesses in foodborne disease surveillance systems. In many parts of the world, especially in Africa and Asia, pathogens such as enterotoxigenic and enterohemorrhagic Escherichia coli, Shigella spp., Salmonella spp., enterotoxigenic Staphylococcus aureus, Listeria monocytogenes, and Bacillus cereus have been detected in fermented foods. Therefore, this review, in addition to the positive aspects, presents the potential risk associated with the consumption of this type of products.

Flies as a potential vector of selected alert pathogens in a hospital environment.

Multi-drug resistant pathogens are a global problem. Flies are a potential vector of multi-drug resistant pathogens, which can be particularly dangerous in the hospital environment. This study aimed to evaluate flies as vectors of alert pathogens. The research material consisted of 100 flies (Musca domestica (46.0%), Lucilia sericata (28.0%), and Calliphora vicina (26.0%)) collected at the University Hospital No. 1 dr. A. Jurasz in Bydgoszcz (Poland) in 2018-2019 (summer months). The presence of bacteria of the genera: Enterococcus, Staphylococcus, Escherichia, Leclercia, Citrobacter, Hafnia, Providencia, Proteus, Enterobacter, Klebsiella, Raoultella, Morganella, Moellerella, Bordetella, Pantoea, Serratia, Plesiomonas, Wohlfahrimonas, and Lelliottia was confirmed. The most frequently isolated species included: Enterococcus faecalis (n = 64), Escherichia coli (n = 43) and Moellerella wisconsensis (n = 24). The infection rate and antibiotic resistance of bacteria were assessed. One strain of Proteus mirabilis (isolated from Calliphora vicina) produced ESBLs (extended-spectrum beta-lactamases). The infection rate was 0.38%, 0.26%, and 0.20% for Musca domestica, Lucilia sericata, and Calliphora vicina, respectively. The flies from a hospital area were not a vector of alert pathogens. Monitoring flies as potential vectors of pathogens is an important aspect of public health, especially for hospitalized patients.

The Differences in the Level of Anti-SARS-CoV-2 Antibodies after mRNA Vaccine between Convalescent and Non-Previously Infected People Disappear after the Second Dose-Study in Healthcare Workers Group in Poland.

(1) Background: In many infections, antibodies play a crucial role in controlling infection. In COVID-19, the dynamics of the immune system response to SARS-CoV-2 is not fully understood. (2) Methods: The study was conducted on 120 healthcare workers from Dr. Antoni Jurasz University Hospital No. 1 in Bydgoszcz, between June and December 2020. In all participants, IgA and IgG antibody serum concentrations were measured using the semi-quantitative Anti-SARS-CoV-2 ELISA test (Euroimmun). After vaccination, in January and February 2021, antibody levels were examined using the quantitative IgG Anti-SARS-CoV-2 Quantivac ELISA test (Euroimmun). (3) Results: During the whole study period, the SARS-CoV-2 infection was confirmed in 29 (24.2%) participants. In all infected participants, IgA and IgG antibodies were detectable after infection by semi-quantitative serological tests. Levels of antibodies were higher one month after the first dose in the convalescents than in the non-previously infected participants. In this second group, the level of antibodies increased significantly after the second dose of vaccines compared to the first dose. (4) Conclusions: The level of antibodies after the first dose of vaccine in the convalescents' group is higher than in the SARS-CoV-2 non-infected group, but the differences disappear after the second vaccination.

Adaptive Response of Listeria monocytogenes to the Stress Factors in the Food Processing Environment.

Listeria monocytogenes are Gram-positive, facultatively anaerobic, non-spore-forming bacteria that easily adapt to changing environmental conditions. The ability to grow at a wide range of temperatures, pH, and salinity determines the presence of the pathogen in water, sewage, soil, decaying vegetation, and animal feed. L. monocytogenes is an etiological factor of listeriosis, especially dangerous for the elderly, pregnant women, and newborns. The major source of L. monocytogenes for humans is food, including fresh and smoked products. Its high prevalence in food is associated with bacterial adaptation to the food processing environment (FPE). Since the number of listeriosis cases has been progressively increasing an efficient eradication of the pathogen from the FPE is crucial. Understanding the mechanisms of bacterial adaptation to environmental stress will significantly contribute to developing novel, effective methods of controlling L. monocytogenes in the food industry.

SARS-CoV-2-Morphology, Transmission and Diagnosis during Pandemic, Review with Element of Meta-Analysis.

The outbreak of Coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome (SARS) coronavirus 2 (SARS-CoV-2). Thus far, the virus has killed over 2,782,112 people and infected over 126,842,694 in the world (state 27 March 2021), resulting in a pandemic for humans. Based on the present data, SARS-CoV-2 transmission from animals to humans cannot be excluded. If mutations allowing breaking of the species barrier and enhancing transmissibility occurred, next changes in the SARS-CoV-2 genome, leading to easier spreading and greater pathogenicity, could happen. The environment and saliva might play an important role in virus transmission. Therefore, there is a need for strict regimes in terms of personal hygiene, including hand washing and surface disinfection. The presence of viral RNA is not an equivalent of active viral infection. The positive result of the RT-PCR method may represent either viral residues or infectious virus particles. RNA-based tests should not be used in patients after the decline of disease symptoms to confirm convalescence. It has been proposed to use the test based on viral, sub-genomic mRNA, or serological methods to find the immune response to infection. Vertical transmission of SARS-CoV-2 is still a little-known issue. In our review, we have prepared a meta-analysis of the transmission of SARS-CoV-2 from mother to child depending on the type of delivery. Our study indicated that the transmission of the virus from mother to child is rare, and the infection rate is not higher in the case of natural childbirth, breastfeeding, or contact with the mother. We hope that this review and meta-analysis will help to systemize knowledge about SARS-CoV-2 with an emphasis on diagnostic implications and transmission routes, in particular, mother-to-child transmission.

Human Skin Microbiome: Impact of Intrinsic and Extrinsic Factors on Skin Microbiota.

The skin is the largest organ of the human body and it protects the body from the external environment. It has become the topic of interest of researchers from various scientific fields. Microorganisms ensure the proper functioning of the skin. Of great importance, are the mutual relations between such microorganisms and their responses to environmental impacts, as dysbiosis may contribute to serious skin diseases. Molecular methods, used for microorganism identification, allow us to gain a better understanding of the skin microbiome. The presented article contains the latest reports on the skin microbiota in health and disease. The review discusses the relationship between a properly functioning microbiome and the body's immune system, as well as the impact of internal and external factors on the human skin microbiome.

SARS-CoV-2 in the environment-Non-droplet spreading routes.

The new coronavirus SARS-CoV-2, first identified in Wuhan (China) in December 2019, represents the same family as the Serve Acute Respiratory Syndrome Coronavirus-1 (SARS-CoV-1). These viruses spread mainly via the droplet route. However, during the pandemic of COVID-19 other reservoirs, i.e., water (surface and ground), sewage, garbage, or soil, should be considered. As the infectious SARS-CoV-2 particles are also present in human excretions, such a non-droplet transmission is also possible. A significant problem is the presence of SARS-CoV-2 in the hospital environment, including patients' rooms, medical equipment, everyday objects and the air. Relevant is selecting the type of equipment in the COVID-19 hospital wards on which the virus particles persist the shortest or do not remain infectious. Elimination of plastic objects/equipment from the environment of the infected person seems to be of great importance. It is particularly relevant in water reservoirs contaminated with raw discharges. Wastewater may contain coronaviruses and therefore there is a need for expanding Water-Based Epidemiology (WBE) studies to use obtained values as tool in determination of the actual percentage of the SARS-CoV-2 infected population in an area. It is of great importance to evaluate the available disinfection methods to control the spread of SARS-CoV-2 in the environment. Exposure of SARS-CoV-2 to 65-70% ethanol, 0.5% hydrogen peroxide, or 0.1% sodium hypochlorite has effectively eliminated the virus from the surfaces. Since there are many unanswered questions about the transmission of SARS-CoV-2, the research on this topic is still ongoing. This review aims to summarize current knowledge on the SARS-CoV-2 transmission and elucidate the viral survival in the environment, with particular emphasis on the possibility of non-droplet transmission.

Genetic Diversity and Distribution of Virulence-Associated Genes in Y. enterocolitica and Y. enterocolitica-Like Isolates from Humans and Animals in Poland.

Yersinia enterocolitica, widespread within domestic and wild-living animals, is a foodborne pathogen causing yersiniosis. The goal of this study was to assess a genetic similarity of Y. enterocolitica and Y. enterocolitica-like strains isolated from different hosts using Multiple Locus Variable-Number Tandem Repeat Analysis (MLVA) and Pulsed-Field Gel Electrophoresis (PFGE) methods, and analyze the prevalence of virulence genes using multiplex-Polymerase Chain Reaction (PCR) assays. Among 51 Yersinia sp. strains 20 virulotypes were determined. The most common virulence genes were ymoA, ureC, inv, myfA, and yst. Yersinia sp. strains had genes which may contribute to the bacterial invasion and colonization of the intestines as well as survival in serum. One wild boar Y. enterocolitica 1A strain possessed ail gene implying the possible pathogenicity of 1A biotype. Wild boar strains, represented mainly by 1A biotype, were not classified into the predominant Variable-Number Tandem Repeats (VNTR)/PFGE profile and virulotype. There was a clustering tendency among VNTR/PFGE profiles of pig origin, 4/O:3, and virulence profile. Pig and human strains formed the most related group, characterized by ~80% of genetic similarity what suggest the role of pigs as a potential source of infection for the pork consumers.

Assessment of the Prevalence and Drug Susceptibility of Listeria monocytogenes Strains Isolated from Various Types of Meat.

Listeria monocytogenes are the etiological factor of listeriosis, and their main source for humans is food. The aim of the current study was to assess the contamination of various types of meat and the drug susceptibility of isolated L. monocytogenes. Between 2016-2018, 6000 swabs were taken (2000 annually) from the surface of pork, beef, and poultry. The analysis of intermediate and finished product samples was carried out in accordance with ISO 11290-1 (International Organization for Standardization). The genetic similarity assessment of the isolates obtained was based on the Pulsed Field Gel Electrophoresis (PFGE) method, and drug-sensitivity assessment using the disc-diffusion method. We found 2.1% of collected samples were L. monocytogenes positive. The level of meat contamination varied depending on its matrix. Most L. monocytogenes were isolated from poultry. It was shown that 39 (32.5%) strains were sensitive to all tested antibiotics and eight (6.7%) were resistant to all five tested antimicrobials. Most strains tested were resistant to cotrimoxazole (55; 45.8%) and meropenem (52; 43.3%), followed by erythromycin (48; 40.0%), penicillin (31; 25.8%), and ampicillin (21; 17.5%). High prevalence of this pathogen may be a serious problem, especially when linked with antibiotic resistance and high percentage of serotypes responsible for listeriosis outbreaks.

Ascorbic Acid Changes Growth of Food-Borne Pathogens in the Early Stage of Biofilm Formation.

Since bacterial biofilm may contribute to the secondary contamination of food during the manufacturing/processing stage there is a need for new methods allowing its effective eradication. Application of food additives such as vitamin C already used in food industry as antioxidant food industry antioxidants may be a promising solution. The aim of this research was evaluation of the impact of vitamin C (ascorbic acid), in a range of concentrations 2.50 µg mL-1-25.0 mg mL-1, on biofilms of Staphylococcus aureus, Escherichia coli, and Listeria monocytogenes strains isolated from food. The efficacy of ascorbic acid was assessed based on the reduction of optical density (λ = 595 nm). The greatest elimination of the biofilm was achieved at the concentration of vitamin C of 25.0 mg mL-1. The effect of the vitamin C on biofilm, however, was strain dependent. The concentration of 25.0 mg mL-1 reduced 93.4%, 74.9%, and 40.5% of E. coli, L. monocytogenes, and S. aureus number, respectively. For E. coli and S. aureus lower concentrations were ineffective. In turn, for L. monocytogenes the biofilm inhibition was observed even at the concentration of 0.25 mg mL-1. The addition of vitamin C may be helpful in the elimination of bacterial biofilms. Nonetheless, some concentrations can induce growth of the pathogens, posing risk for the consumers' health.