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Epstein–Barr virus (EBV) is an oncogenic virus classified by the World Health Organization as a class 1 carcinogen. Post-transplant lymphoproliferative disorders are believed to be strongly related to an EBV infection. Monitoring of EBV DNAemia is recommended to assess the risk of reactivation of latent infection and to assess the effectiveness of therapy. Currently, various types of clinical specimens are used for this purpose. The aim of the study was to assess a reliable method of EBV viral load investigation depending on the clinical material used: whole blood or plasma samples. We found that of 134 EBV-DNA-positive whole-blood samples derived from 51 patients (mostly hemato-oncology or post-transplantation), only 43 (32.1%) were plasma-positive. Of these, 37 (86.0%) had lower plasma DNAemia compared to the corresponding whole-blood samples. We conclude that whole-blood samples have a higher sensitivity than plasma samples in EBV DNA detection. The clinical utility of the tests is unclear, but our results suggest that either whole blood or plasma should be used consistently for EBV viral load monitoring.

BackgroundListeria monocytogenesare Gram-positive rods, which are the etiological factor oflisteriosis.L. monocytogenesquickly 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 ofL. monocytogenesstrains isolated from food products and clinical material.ResultsWe distinguished fifty genetically different strains ofL. 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 × 108PFU. 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.ConclusionsData 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.

ObjectiveMicrobial infection plays an important role in exacerbation of chronic otitis media. The aim of this study was to analyse the microbiota in chronic otitis media in the context of local treatment.MethodIn this prospective study, samples for microbiological examination were taken from 119 patients who underwent operation because of chronic otitis media.ResultsThe results were compared between groups depending on the type of operation (none, tympanoplasty or radical), the presence of cholesteatoma or granulomatous tissue or discharge from the ear as a symptom of exacerbation. Antibiotic susceptibility of germs was analysed to define the strategy of treatment. A total of 209 samples were collected from 119 patients with chronic otitis media.ConclusionPseudomonas aeruginosa and Staphylococcus aureus were pathogens most frequently identified from the ear in the course of chronic otitis media. Pseudomonas aeruginosa was concerned with major pathology of the middle ear (radical surgery, cholesteatoma or granulomatous tissue, persisting discharge after treatment), whereas Staphylococcus aureus was obtained in dry perforations without other pathology in the middle-ear cavity. Ciprofloxacin was effective against Staphylococcus aureus, but Pseudomonas aeruginosa strains were ciprofloxacin resistant.

Urinary Tract Infections (UTIs) are common outpatient and inpatient infections, often treated with empirical therapy.

Cefiderocol (CFDC) is a novel, broad-spectrum siderophore cephalosporin with potential activity against multi-drug (MDR) and extensively drug-resistant (XDR) Enterobacterales, including carbapenem-resistant strains. We assessed the in vitro susceptibility to CFDC of MDR, and XDR

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.

Enterobacterales as opportunistic pathogens are commonly associated with nosocomial infections. With increasing frequency, Gram-negative bacilli, especially Klebsiella pneumoniae strains, are mul- tidrug-resistant or pandrug-resistant. Carbapenems were used as the drugs of choice for the treat- ment of infections caused by multidrug-resistant Gram-negative bacilli. The aim of this study was to assess the usefulness of the RESIST-4 O.K.N.V. K-SeT for the rapid detection and identification of the most important carbapenemases (OXA-48, KPC, NDM, VIM) in Enterobacterales bacilli. The study involved the isolates of 97 Enterobacterales strains. The ability to produce carbapenemases was determined by the immunochromatographic RESIST-4 O.K.N.V. K-SeT test. This test detected carbapenemases OXA-48, KPC, NDM, and VIM. For the RESIST-4 O.K.N.V. K-SeT test, a positive result was obtained for 93 strains (95.9%). Four strains negative in the RESIST-4 O.K.N.V. K-SeT were positive in the Eazyplex®SuperBugCRE and PCR. These strains produce VIM enzymes. RE- SIST-4 O.K.N.V. K-SeT test is rapid, simple to perform and can be used for fast detection of the most important carbapenemases (OXA-48, KPC, NDM, VIM) among Gram-negative bacilli.

Three

Introduction: The normal vaginal microbiota (mainly Lactobacillus spp.) affects the health of these areas. Bacterial vaginosis is a serious health problem among many women, especially dangerous for pregnant women. The study aimed to assess the impact of Lactobacillus spp. strains on the population of Listeria monocytogenes isolated from women. Materials and methods: The research material consisted of reference strains of Lactobacillus spp.: L. acidophilus (LAC), L. fermentum (LFE), L. gasseri (LGA), L. plantarum (LPL), the strain L. monocytogenes ATCC 19111 and 7 L. monocytogenes strains isolated from the vagina. Results: The highest antagonistic activity was shown for the mixed culture of all Lactobacillus strains (LACTO MIX) used in the experiment. Among the individual strains of Lactobacillus spp. strains, L. plantarum turned out to most effectively reduce L. monocytogenes number (reduction of 5.74 log CFU × ml-1). The least effective in inhibiting the growth of L. monocytogenes was the L. acidophilus strain (reduction of L. monocytogenes of a number of 2.21 log CFU × ml-1). Conclusions: The presence of Lactobacillus spp. in the genital tract limits the development of bacterial infections, which is an important aspect especially for pregnant women.

Objectives: This study evaluated the diagnostic performance of the eazyplex® SuperBug CRE (eSBCRE) system, based on a loop-mediated isothermal amplification (LAMP), for the detection of the most co...

Wstęp Bakterie Gram-ujemne są istotnym czynnikiem etiologicznym szerokiego zakresu zakażeń szpitalnych. Rozpowszechnienie szczepów bakterii wielolekoopornych, zarówno należących do rzędu Enterobacterales, jak i niefermentujących pałeczek z gatunku Pseudomonas aeruginosa, stanowi poważne wyzwanie dla współczesnej medycyny. Badanie nowych opcji terapeutycznych oraz monitorowanie antybiotykowrażliwości drobnoustrojów jest ważnym elementem działań zapobiegających rozwojowi i utrzymywaniu się zakażeń z udziałem szczepów wielolekoopornych. Celem pracy było porównanie aktywności in vitro połączenia ceftazydymu z awibaktamem z najczęściej stosowanymi w Polsce antybiotykami, mającymi zastosowanie w leczeniu zakażeń bakteriami Gram-ujemnymi z rzędu Enterobacterales oraz P. aeruginosa, w ramach programu badawczego ATLAS. Materiał i metody W pracy analizowano wrażliwość 1677 szczepów rzędu Enterobacterales oraz 500 niefermentujących pałeczek P. aeruginosa, izolowanych w latach 2014–2018 w ośrodkach biorących udział w programie, na ceftazydym z awibaktamem oraz inne antybiotyki stosowane w leczeniu zakażeń pałeczkami Gram-ujemnymi. Wyniki Niemal 99% (98,9%) szczepów rzędu Enterobacterales, w tym 96,9% szczepów wytwarzających beta-laktamazy typu ESBL, było wrażliwych na ceftazydym z awibaktamem. Notowano wysoki odsetek szczepów Escherichia coli oraz Klebsiella pneumoniae wrażliwych na ceftazydym z awibaktamem, odpowiednio: 99,8% i 98,9%, w tym także szczepów ESBL-dodatnich tych samych gatunków, odpowiednio: 98,5% i 98,7%. Wrażliwość szczepów P. aeruginosa na ceftazydym z awibaktamem była niższa niż badanych szczepów Enterobacterales, niemniej jednak kształtowała się na poziomie 91,7%, co czyniło ceftazydym z awibaktamem drugim, po kolistynie, najlepiej działającym antybiotykiem wobec tych bakterii. Wnioski Przedstawione dane w połączeniu z dotychczas opublikowanymi wynikami farmakokinetycznymi, farmakodynamicznymi, klinicznymi i bezpieczeństwa wskazują, że połączenie ceftazydymu z awibaktamem może stanowić cenną opcję leczenia zakażeń wywoływanych przez Enterobacterales oraz P. aeruginosa, w tym zakażeń szczepami wielolekoopornymi.

Aim: Escherichia coli is one of the Gram-negative bacteria, known to cause many nosocomial infections. Multi-drug (MDR) and extensively-drug resistant (XDR). E. coli are of particular note, due to significant limitations in antibiotic therapy. Ceftolozane-tazobactam and ceftazidime-avibactam are novel therapeutic options against Gram-negative bacteria; hence the aim of this study was to evaluate and compare the in vitro activity of ceftolozane-tazobactam and ceftazidime-avibactam against MDR and XDR clinical E. coli isolates. Material/Methods: The study included 100 non-replicate E. coli isolates derived from clinical samples of patients hospitalized in teaching hospitals. Bacteria were identified by applying mass spectrometry in the MALDI Biotyper system (Bruker). ESBL (blaCTX-M-1group, blaCTX-M-9group) and carbapenemase (blaKPC, blaVIM, blaNDM, blaOXA-48, blaOXA-181) genes were detected using the eazyplex® SuperBug CRE test, based on a loop-mediated isothermal amplification (LAMP). The in vitro susceptibility to ceftolozane-tazobactam and ceftazidime-avibactam was tested using validated MIC Test strips (Liofilchem). Results: All 84 extended-spectrum β-lactamase-producing (ESBL) E. coli isolates were susceptible to ceftazidime-avibactam and 83 to ceftolozane-tazobactam. Among 17 E. coli isolates with resistance to at least one of the carbapenems, three (17.6%) were susceptible to ceftolozane-tazobactam and ceftazidime-avibactam. All 14 blaVIM gene-positive E. coli isolates were resistant to both ceftolozane-tazobactam and ceftazidime-avibactam. Both antibiotics were active against blaCTX-M-9group and blaOXA-48 gene-positive E. coli isolates, but they were not active against blaCTX-M-1group and blaVIM gene-positive isolates. Conclusions: Ceftolozane-tazobactam and ceftazidime-avibactam are alternative, non-carbapenem therapeutic options for ESBL-positive E. coli strains, and they are promising in the treatment of carbapenem-resistant E. coli strains, but not for those carrying the metallo-β-lactamase enzymes. Both drug combinations have comparable activity against ESBL, however, lower MIC values were found for ceftazidime-avibactam.

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.

(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.

Antibiotic resistance (AR) and multidrug resistance (MDR) have been confirmed for all major foodborne pathogens: Campylobacter spp., Salmonella spp., Escherichia coli and Listeria monocytogenes. Of great concern to scientists and physicians are also reports of antibiotic-resistant emerging food pathogens—microorganisms that have not previously been linked to food contamination or were considered epidemiologically insignificant. Since the properties of foodborne pathogens are not always sufficiently recognized, the consequences of the infections are often not easily predictable, and the control of their activity is difficult. The bacteria most commonly identified as emerging foodborne pathogens include Aliarcobacter spp., Aeromonas spp., Cronobacter spp., Vibrio spp., Clostridioides difficile, Escherichia coli, Mycobacterium paratuberculosis, Salmonella enterica, Streptocccus suis, Campylobacter jejuni, Helicobacter pylori, Listeria monocytogenes and Yersinia enterocolitica. The results of our analysis confirm antibiotic resistance and multidrug resistance among the mentioned species. Among the antibiotics whose effectiveness is steadily declining due to expanding resistance among bacteria isolated from food are β-lactams, sulfonamides, tetracyclines and fluoroquinolones. Continuous and thorough monitoring of strains isolated from food is necessary to characterize the existing mechanisms of resistance. In our opinion, this review shows the scale of the problem of microbes related to health, which should not be underestimated.

(1) Background: The main source of transmission of

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.

Multidrug-resistant

Lactic acid bacteria belonging to Lactobacillus spp. and Lacticaseibacillus spp. are a natural part of fermented milk and other food products, probiotic supplements and human microbiota. They mainly belong to mucosal microflora, especially oral, vaginal and intestinal. Lacticaseibacillus spp. strains included in probiotics are generally characterised as safe microorganisms, and the species are concerned bacteria with very low pathogenic potential. However, infections caused by Lactobacillus spp. and Lacticaseibacillus spp., including bacteraemia and endocarditis, occur occasionally. The aim of the study was to present two cases of bacteraemia due to Lacticaseibacillus rhamnosus associated with the use of a probiotic product. It afflicted patients in intensive care units. The investigation was preliminarily based on clinical and microbiological recognition of the cases. The initial observation was laboratory confirmed with the application of pulsed-field gel electrophoresis (PFGE) results. Identical PFGE patterns were obtained for the evaluated strains and the strains derived from a commercially available probiotic that was administered to those patients. The increasing number of studies describing opportunistic infections due to probiotic strains of Lacticaseibacillus spp. should result in verifying the safety of probiotic formulations used in immunocompromised patients and forming detailed guidelines for the use of probiotics among patients from several risk groups.

Healthcare-associated infections caused by multidrug-resistant Acinetobacter baumannii strains are a serious global threat. Therefore, it is important to expand the knowledge on the mechanisms of pathogenicity of these particular bacteria. The aim of this study was to assess the distribution of selected virulence factor genes (bap, surA1, omp33-36, bauA, bauS, and pld) among carbapenem-non-susceptible clinical A. baumannii isolates and to evaluate their potential usefulness as genetic markers for rapid diagnostics of A. baumannii infections. Moreover, we aimed to compare the virulence genes prevalence with the occurrence of carbapenemases genes. A total of 100 carbapenem-non-susceptible A. baumannii clinical isolates were included in the study. The presence of virulence factors and blaOXA genes was evaluated by real-time PCR. The occurrence of virulence factors genes was as follows: 100.0% for the bap and surA1 genes, 99.0% for the basD and pld genes. The bauA and omp33-36 genes were absent among the studied strains. The predominant genes (bap and surA1) are involved in biofilm formation and their presence among all clinical strains can be applied as a genetic marker to recognize A. baumannii infection. High frequencies of the basD gene—involved in siderophore biosynthesis and the gene encoding phospholipase D (pld)—were also noted among blaOXA-positive strains, showing their potential role in a pathogenicity of blaOXA-positive A. baumannii clinical strains.

Pseudomonas aeruginosa is an opportunistic pathogen encoding several virulence factors in its genome, which is well-known for its ability to cause severe and life-threatening infections, particularly among cystic fibrosis patients. The organism is also a major cause of nosocomial infections, mainly affecting patients with immune deficiencies and burn wounds, ventilator-assisted patients, and patients affected by other malignancies. The extensively reported emergence of multidrug-resistant (MDR) P. aeruginosa strains poses additional challenges to the management of infections. The aim of this study was to compare the incidence rates of selected virulence-factor-encoding genes and the genotype distribution amongst clinical multidrug-sensitive (MDS) and MDR P. aeruginosa strains. The study involved 74 MDS and 57 MDR P. aeruginosa strains and the following virulence-factor-encoding genes: lasB, plC H, plC N, exoU, nan1, pilA, and pilB. The genotype distribution, with respect to the antimicrobial susceptibility profiles of the strains, was also analyzed. The lasB and plC N genes were present amongst several P. aeruginosa strains, including all the MDR P. aeruginosa, suggesting that their presence might be used as a marker for diagnostic purposes. A wide variety of genotype distributions were observed among the investigated isolates, with the MDS and MDR strains exhibiting, respectively, 18 and 9 distinct profiles. A higher prevalence of genes determining the virulence factors in the MDR strains was observed in this study, but more research is needed on the prevalence and expression levels of these genes in additional MDR strains.

(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.

Among the diseases that pose a serious threat to public health, those caused by viruses are of great importance. The Nipah virus (NiV) belonging to the Paramyxoviridae family was reported in Malaysia in 1998/1999. Due to its high mortality in humans, its zoonotic nature, the possibility of human-to-human transmission, and the lack of an available vaccine, the World Health Organization (WHO) has recognized it as a global health problem. Depending on strain specificity, neurological symptoms and severe respiratory disorders are observed in NiV infection. In most confirmed cases of NiV epidemics, the appearance of the virus in humans was associated with the presence of various animal species, but generally, bats of Pteropus species are considered the most important natural animal NiV reservoir and vector. Consumption of contaminated food, contact with animals, and “human-to-human” direct contact were identified as NiV transmission routes. Due to the lack of vaccines and drugs with proven effectiveness against NiV, treatment of patients is limited to supportive and prophylactic.

The main purpose of micro-organisms elimination from the air and surfaces is to ensure microbiological safety in health care facilities or food production plants. Currently, many disinfection methods are used, both physical, chemical and, increasingly, biological. Scientists seek new solutions with high antimicrobial effectiveness (especially against the drug-resistant strains of bacteria), low production and operating costs, and, above all, the safety of patients and food consumers. The limitation of the methods used so far is primarily the micro-organisms acquire the resistance, mainly to antimicrobial agents. One of the new and alternative methods of disinfection is radiant catalytic ionization (RCI). RCI is an active method of air and surface purification. The technology proved high efficiency against viruses, Gram-positive and -negative bacteria, and fungi, both in the air and on surfaces (planktonic forms and biofilm). RCI has many advantages as well as some minor limitations. This overview summarizes the current knowledge about RCI technology.

BACKGROUND: Listeria monocytogenes is a cause of listeriosis, dangerous especially for elderly, immunocompromised people and pregnant women. Ability to colonize biotic and abiotic surfaces and form biofilm by these pathogens poses a serious threat for the hospitalized, catheterized patients. METHODS: The study was conducted on 29 L. monocytogenes strains isolated from clinical materials (blood, cerebrospinal fluid, swabs from vagina) and the reference strain L. monocytogenes ATCC 1911. Ability to form biofilm in 96-well plates and drug susceptibility (disk diffusion method) of tested strains was determined. RESULTS: All strains formed biofilm though it’s intensity was correlated with source of isolation. The strong biofilm formed 72.73 % of isolates from cerebrospinal fluid ((A570 0.421 – 1.3), 75.0 % of blood isolates 9 (A570 0.389 – 1.063) and 50.0 % of isolates from vaginal swabs (A570 0.457 – 0.487). The strongest biofilm was formed by strains derived from cerebrospinal fluid whereas isolates from vaginal swabs, which strongly formed a biofilm accounted for 50.0% of the studied population (absorbance 0.457 - 0.487). It was found that 93.1 % (n=27) of strains were susceptible to all drugs tested. Two strains (6.9 %) were resistant to cotrimoxazol and 1 strain (3.45 %) to erythromycin. CONCLUSIONS: Diverse ability to form biofilm by clinical L. monocytogenes strains is an important aspect in prophylaxis in catheterized patients.

Introduction. Raoultella spp. representatives are Gram-negative rod-shaped bacteria of the Enterobacteriaceae family. These bacteria are commonly found in the natural environment. Aim. The aim of the study was to indicate the reliable method for Raoultella spp. strains identification, evaluate the susceptibility of Raoultella spp. strains to selected antimicrobials and to detect their resistance mechanisms to beta-lactams. Methodology. Susceptibility of the strains to chosen antimicrobials was determined using the automatic method. The presence of particular antimicrobial resistant mechanism and genes encoding ESBLs and MBLs was determined respectively with double-disc synergy test and commercially available kit – eazyplex SuperBug CRE test (Amplex Diagnostics) and standard PCR. For the selected strains, DNA sequencing was performed. Results. Amongst 105 of the examined Raoultella spp. strains, majority were sensitive to: imipenem (99.0 %), meropenem (98.1 %), gentamicin (93.3 %) and ciprofloxacin (92.4 %). Of the tested Raoultella strains, thirteen (12.4 %) produced ESBLs and one strain simultaneously ESBLs and MBLs. The DNA sequencing results were as follows: for all the reference strains the correct species identification was achieved, for the analysed strains two were identified as R. planticola and one as R. ornithinolytica . Conclusion. Although Raoultella spp. strains remain sensitive to antibiotics, there is a constant need to monitor the sensitivity of these bacteria to selected antimicrobials. Isolation of a multi-drug resistant R. ornithinolytica strain indicates that even the less frequently isolated species of Enterobacteriaceae family should be precisely identified because they might be of clinical importance and the particular strain can also produce enzymes that pose the greatest threat today.

The multi-drug resistance of Gram-negative rods is one of the most important issues of present medicine. In recent years, more and more strains resistant to the majority or to all possible therapeutic options have been isolated—especially Klebsiella spp. and Pseudomonas spp. representatives. It is very important to detect strains with these phenotypes as quickly and reliably as possible. The aim of the study was to evaluate the usefulness of eazyplex® SuperBug CRE test (Amplex Diagnostics) for the detection of the most important beta-lactam resistance genes. eazyplex® SuperBug CRE test is based on the loop-mediated isothermal amplification (LAMP) method, and detects genes for the following beta-lactamases: KPC, NDM-1, VIM, OXA-48, CTX-M1, CTX-M9 and OXA-181. The study involved 87 strains. For all of the positive strains in the LAMP method, additional PCR were performed to increase the spectrum of ESBLs detected by the genes encoding for enzymes belonging to TEM and SHV families. The results obtained by the tested method and standard PCR were consistent for all Klebsiella spp. strains. The discrepancy between the evaluated test and PCR results was observed for one P. aeruginosa strain. The eazyplex® SuperBug CRE test can be used for quick detection of the most important beta-lactam resistance mechanisms amongst Gram-negative rods.

Due to increasing antibiotic resistance Klebsiella pneumoniae is a serious threat for the hospitalized patients. The aim of this study was the assessment of radiant catalytic ionization (RCI) efficacy on K. pneumoniae reduction in the air and on selected surfaces. Four K. pneumoniae NDM and ESBLs-producing strains were included in the study. Three types of surface were tested: cotton-polyester, terry and PVC. It was found that RCI significantly reduced the number of bacteria from all types of surface (terry: 0·56-1·22 log CFU m2 , cotton-polyester: 2·15-3·71 log CFU per m2 , PVC: 4·45-4·92 log CFU per m2 ) as well as from the air (1·80 log CFU per m3 ). The RCI technology may be a useful disinfection method in hospitals. SIGNIFICANCE AND IMPACT OF THE STUDY: Microbial contamination of air and surfaces in hospitals play an important role in healthcare-associated infections. The aim was the assessment of Klebsiella pneumoniae elimination using radiant catalytic ionization (RCI). K. pneumoniae are aetiological agent of nosocomial infections, such as: pneumonia, infections of urinary tract, blood, e.t.c. The strains producing the New Delhi metallo-β-lactamases are one of the greatest epidemiological threat. The use of RCI eliminate the tested bacteria from the hospital environment, but can also be effective in food processing plants or public facilities, ensuring the safety of people and products. This research is scarce in references and has a large innovation and application potential.

Listeria monocytogenes is the etiological factor of listeriosis. The main source of these organisms is food, including dairy products. The aim was to determine the multiple correlations between the drug susceptibility, virulence genes (VGs), and biofilm formation on silicone teat cups of milk-borne and human L. monocytogenes strains. The spread of L. monocytogenes via contaminated teat rubbers was assessed. The L. monocytogenes strains recovered from milk (18), human blood (10), and the reference strain ATCC®19111™ were used in the study. Penicillin resistance was the most prevalent resistance in the milk isolates (n=8; 44.4%), whereas among clinical strains erythromycin resistance was predominating - (n=6; 60%). The most frequent VGs among strains isolated from milk were hlyA (100%) and plcB (100%) whereas in strains isolated from blood - hlyA (100%) and prfA (90%). All tested VGs were present in 50% of blood isolates and 11% of milk-borne strains. The strains isolated from milk formed a significantly stronger biofilm. The strains with more numerous virulence genes were resistant to more antibiotics and formed a stronger biofilm. It was shown that contaminated teat cups might contribute to the transmission of L. monocytogenes in the herd. It seems reasonable to monitor the occurrence of L. monocytogenes biofilm in a dairy processing environment. Listeria monocytogenes is the etiological factor of listeriosis. The main source of these organisms is food, including dairy products. The aim was to determine the multiple correlations between the drug susceptibility, virulence genes (VGs), and biofilm formation on silicone teat cups of milk-borne and human L. monocytogenes strains. The spread of L. monocytogenes via contaminated teat rubbers was assessed. The L. monocytogenes strains recovered from milk (18), human blood (10), and the reference strain ATCC®19111™ were used in the study. Penicillin resistance was the most prevalent resistance in the milk isolates (n=8; 44.4%), whereas among clinical strains erythromycin resistance was predominating – (n=6; 60%). The most frequent VGs among strains isolated from milk were hlyA (100%) and plcB (100%) whereas in strains isolated from blood – hlyA (100%) and prfA (90%). All tested VGs were present in 50% of blood isolates and 11% of milk-borne strains. The strains isolated from milk formed a significantly stronger biofilm. The strains with more numerous virulence genes were resistant to more antibiotics and formed a stronger biofilm. It was shown that contaminated teat cups might contribute to the transmission of L. monocytogenes in the herd. It seems reasonable to monitor the occurrence of L. monocytogenes biofilm in a dairy processing environment.

Background: Listeria monocytogenes is a Gram-positive, foodborne pathogen. Biofilms formed by this bacterium are a serious problem in the food industry. Bacteria in biofilms are much more resistant to cleaning and disinfection agents posing a risk of food recontamination. The aim of this study was the assessment of the influence of initial sonication on disinfectant efficacy, based on QAC, against L. monocytogenes biofilm on the stainless steel. Methods: The biofilm formed on the stainless steel by the reference strain L. monocytogenes ATCC 19111 was sonicated for 1 and 5 minutes (500W/ 20kHz/ 100% amplitude). Then disinfection with quaternary ammonium compounds (0.5% working solution) was applied for 1 and 5 minutes and the number of bacteria recovered from the biofilm was assessed. Results: It was found that disinfection was more efficient than sonication (p ≤ 0.05). However, the combination of sonication and disinfection significantly improved biofilm eradication compared to the use of one of these methods separately (p ≤ 0.05). The greatest reduction of bacteria number was achieved after 5 minutes of sonication combined with 5 minutes of disinfection (6.42 log CFU × cm-2), whereas the lowest reduction was observed after 1 minute-sonication (2.03 log CFU × cm-2). Conclusions: Combination of sonication and disinfection based on quaternary ammonium compounds is an effective method allowing biofilm eradication from the food production surfaces.

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.

The aim of this study was to evaluate the usefulness of the Accelerate Pheno™ system (APS) (Accelerate Diagnostics, Denver, CO, USA) for rapid laboratory diagnosis of bloodstream infections. The study included 45 positive blood samples obtained from patients hospitalized in University Hospital No. 1 in Bydgoszcz, Poland. In 40 (88.9%) blood samples, the APS was capable of identification of at least one microorganism at the genus or species level and in 38 (84.4%) of them additionally assessed antimicrobial susceptibility. The time of identification and the time to result of antimicrobial susceptibility ranged from 1:32 to 1:42 and 5:02 to 5:36 h, respectively. Six positive blood samples revealed a poly-microbial culture. In these cases, only one out of two or three microorganisms was detected by the APS, and the system assessed antimicrobial susceptibility only for them. For 78.6% positive blood samples, agreement on identification compared to mass spectrometry was found. For all but one sample, a 96–100% compliance of the resistance category was achieved when comparing the antimicrobial susceptibility testing results to conventional methods. Using the APS, the total time to report was reduced from 13:34 to even 63:47 h compared to the standard microbiological laboratory workflow. The APS is a very useful system, especially for the rapid assessment of antimicrobial susceptibility of bacteria directly from positive blood samples, offering the greatest potential for microbiology laboratories operating around the clock.

Viral infections, or their reactivations, are one of the most important groups of transplantation complications that can occur among recipients of both hematopoietic cells and solid organ transplants. They are the most commonly caused by cytomegalovirus (CMV). Currently, the use of whole blood or plasma samples is recommended for CMV viral load monitoring. The aim of the study was to assess and compare the level of CMV DNA, depending on the type of clinical material—whole blood or plasma fraction derived from the same patient. The studies were carried out on 156 whole blood samples in which the presence of CMV genetic material was confirmed and the corresponding plasma samples from the same rounds of sampling. CMV DNA was not present in 59 (37.8%) of plasma samples compared to whole blood-positive counterparts. Of the samples positive in both types of clinical specimen, 77 (79.4%) had higher viral DNA levels in the whole blood samples. There were statistically significant differences in the detected CMV DNA load in the whole blood compared to plasma fraction counterparts (p < 0.001). The detected CMV DNA value is usually higher in whole blood compared to plasma samples of the same patient. Due to the variability in CMV viral load depending on the clinical material used for a particular patient, one type of specimen should be always used consequently for CMV viremia monitoring.

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.

Non-fermenting Gram-negative rods are one of the most commonly isolated bacteria from human infections. These microorganisms are typically opportunistic pathogens that pose a serious threat to public health due to possibility of transmission in the human population. Resistance to beta-lactams, due to carbapenemases synthesis, is one of the most important antimicrobial resistance mechanisms amongst them. The aim of this study was to evaluate the usefulness of the Carbapenem Inactivation Method (CIM), and its modifications, for the detection of carbapenemase activity amongst non-fermenting Gram-negative rods. This research involved 81 strains of Gram-negative rods. Of the tested strains, 55 (67.9%) synthesized carbapenemases. For non-fermenting rods, 100% sensitivity and specificity was obtained in the version of the CIM test using imipenem discs and E. coli ATCC 25922 strain. The CIM test allows for differentiation of carbapenems resistance mechanisms resulting from carbapenemase synthesis from other resistance types. It is a reliable diagnostic method for the detection of carbapenemase activity amongst non-fermenting Gram-negative rods. Application of imipenem discs and P. aeruginosa ATCC 27853 reference strain increases CIM results sensitivity, while imipenem discs and E. coli ATCC 25922 strain use maintains full precision of the test for non-fermenting rods.

Staphylococcus aureus is one of the predominant bacteria isolated from skin and soft tissue infections and a common cause of bloodstream infections. The aim of this study was to compare the rate of resistance to various antimicrobial agents and virulence patterns in a total of 200 S. aureus strains isolated from patients with bacteremia and chronic wounds. Disk diffusion assay and in the case of vancomycin and teicoplanin-microdilution assay, were performed to study the antimicrobial susceptibility of the isolates. The prevalence of genes encoding six enterotoxins, two exfoliative toxins, the Panton–Valentine leukocidin and the toxic shock syndrome toxin was determined by PCR. Of the 100 blood strains tested, the highest percentage (85.0%, 31.0%, and 29.0%) were resistant to benzylpenicillin, erythromycin and clindamycin, respectively. Out of the 100 chronic wound strains, the highest percentage (86.0%, 32.0%, 31.0%, 31.0%, 30.0%, and 29.0%) were confirmed as resistant to benzylpenicillin, tobramycin, amikacin, norfloxacin, erythromycin, and clindamycin, respectively. A significantly higher prevalence of resistance to amikacin, gentamicin, and tobramycin was noted in strains obtained from chronic wounds. Moreover, a significant difference in the distribution of sea and sei genes was found. These genes were detected in 6.0%, 46.0% of blood strains and in 19.0%, and 61.0% of wound strains, respectively. Our results suggest that S. aureus strains obtained from chronic wounds seem to be more often resistant to antibiotics and harbor more virulence genes compared to strains isolated from blood.

Staphylococcus aureus is one of the predominant bacteria isolated from skin and soft tissue infections and a common cause of bloodstream infections. The aim of this study was to compare the rate of resistance to various antimicrobial agents and virulence patterns in a total of 200 S. aureus strains isolated from patients with bacteremia and chronic wounds. Disk diffusion assay and in the case of vancomycin and teicoplanin-microdilution assay, were performed to study the antimicrobial susceptibility of the isolates. The prevalence of genes encoding six enterotoxins, two exfoliative toxins, the Panton–Valentine leukocidin and the toxic shock syndrome toxin was determined by PCR. Of the 100 blood strains tested, the highest percentage (85.0%, 31.0%, and 29.0%) were resistant to benzylpenicillin, erythromycin and clindamycin, respectively. Out of the 100 chronic wound strains, the highest percentage (86.0%, 32.0%, 31.0%, 31.0%, 30.0%, and 29.0%) were confirmed as resistant to benzylpenicillin, tobramycin, amikacin, norfloxacin, erythromycin, and clindamycin, respectively. A significantly higher prevalence of resistance to amikacin, gentamicin, and tobramycin was noted in strains obtained from chronic wounds. Moreover, a significant difference in the distribution of sea and sei genes was found. These genes were detected in 6.0%, 46.0% of blood strains and in 19.0%, and 61.0% of wound strains, respectively. Our results suggest that S. aureus strains obtained from chronic wounds seem to be more often resistant to antibiotics and harbor more virulence genes compared to strains isolated from blood.

Non-fermenting Gram-negative rods are one of the most commonly isolated bacteria from human infections. These microorganisms are typically opportunistic pathogens that pose a serious threat to public health due to possibility of transmission in the human population. Resistance to beta-lactams, due to carbapenemases synthesis, is one of the most important antimicrobial resistance mechanisms amongst them. The aim of this study was to evaluate the usefulness of the Carbapenem Inactivation Method (CIM), and its modifications, for the detection of carbapenemase activity amongst non-fermenting Gram-negative rods. This research involved 81 strains of Gram-negative rods. Of the tested strains, 55 (67.9%) synthesized carbapenemases. For non-fermenting rods, 100% sensitivity and specificity was obtained in the version of the CIM test using imipenem discs and E. coli ATCC 25922 strain. The CIM test allows for differentiation of carbapenems resistance mechanisms resulting from carbapenemase synthesis from other resistance types. It is a reliable diagnostic method for the detection of carbapenemase activity amongst non-fermenting Gram-negative rods. Application of imipenem discs and P. aeruginosa ATCC 27853 reference strain increases CIM results sensitivity, while imipenem discs and E. coli ATCC 25922 strain use maintains full precision of the test for non-fermenting rods.

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 (

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.

The share of Klebsiella pneumoniae in infections has been recently increasing. Multidrug-resistant strains that produce more than one antibiotic resistance mechanism are also increasingly isolated. Contamination of the organs preservation fluid occurs quite often, but the isolated microorganisms are mainly saprophytic bacteria that are part of the skin microbiota (coagulase-negative Staphylococcus, Corynebacterium spp). The following case describes a K. pneumoniae blood infection in a patient after liver transplantation. Susceptibility of the strains to chosen antimicrobials was determined using the automated method. For strain isolated from blood, it was confirmed by loop-mediated isothermal amplification of genetic material.

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.