High Prevalence of Plasmid-Mediated Quinolone Resistance among ESBL/AmpC-Producing Enterobacterales from Free-Living Birds in Poland.

In this study, we investigated the occurrence of plasmid-mediated quinolone resistance (PMQR) in extended-spectrum ß-lactamase- (ESBL) and/or AmpC-type ß-lactamase-producing Enterobacterales isolates from free-living birds in Poland. The prevalence of the qnrB19 gene was 63%, and the distribution of isolates in terms of bacterial species was as follows: 67% (22/33) corresponded to Escherichia coli, 83% (5/6) to Rahnella aquatilis, 44% (4/9) to Enterobacter cloacae and 33% (1/3) to Klebsiella pneumoniae. The qnrB19 gene was also found in a single isolate of Citrobacter freundii. The molecular characteristics of qnrB19-positive isolates pointed to extended-spectrum beta lactamase CTX-M as the most prevalent one (89%) followed by TEM (47%), AmpC (37%) and SHV (16%). This study demonstrates the widespread occurrence of PMQR-positive and ESBL/AmpC-producing Enterobacterales isolates in fecal samples from wild birds. In this work, plasmid pAM1 isolated from Escherichia coli strain SN25556 was completely sequenced. This plasmid is 3191 nucleotides long and carries the qnrB19 gene, which mediates decreased susceptibility to quinolones. It shares extensive homology with other previously described small qnrB19-harboring plasmids. The nucleotide sequence of pAM1 showed a variable region flanked by an oriT locus and a Xer recombination site. The presence of a putative recombination site was detected, suggesting that interplasmid recombination events might have played a role in the development of pAM1. Our results highlight the broad geographical spread of ColE-type Qnr resistance plasmids in clinical and environmental isolates of Enterobacterales. As expected from the results of phenotypic susceptibility testing, no resistance genes other than qnrB19 were identified.

A Novel Cryptic Clostridial Peptide That Kills Bacteria by a Cell Membrane Permeabilization Mechanism.

This work reports detailed characteristics of the antimicrobial peptide Intestinalin (P30), which is derived from the LysC enzyme of Clostridium intestinale strain URNW. The peptide shows a broader antibacterial spectrum than the parental enzyme, showing potent antimicrobial activity against clinical strains of Gram-positive staphylococci and Gram-negative pathogens and causing between 3.04 ± 0.12 log kill for Pseudomonas aeruginosa PAO1 and 7.10 ± 0.05 log kill for multidrug-resistant Acinetobacter baumannii KPD 581 at a 5 µM concentration. Moreover, Intestinalin (P30) prevents biofilm formation and destroys 24-h and 72-h biofilms formed by Acinetobacter baumannii CRAB KPD 205 (reduction levels of 4.28 and 2.62 log CFU/mL, respectively). The activity of Intestinalin is combined with both no cytotoxicity and little hemolytic effect against mammalian cells. The nuclear magnetic resonance and molecular dynamics (MD) data show a high tendency of Intestinalin to interact with the bacterial phospholipid cell membrane. Although positively charged, Intestinalin resides in the membrane and aggregates into small oligomers. Negatively charged phospholipids stabilize peptide oligomers to form water- and ion-permeable pores, disrupting the integrity of bacterial cell membranes. Experimental data showed that Intestinalin interacts with negatively charged lipoteichoic acid (logK based on isothermal titration calorimetry, 7.45 ± 0.44), causes membrane depolarization, and affects membrane integrity by forming large pores, all of which result in loss of bacterial viability. IMPORTANCE Antibiotic resistance is rising rapidly among pathogenic bacteria, becoming a global public health problem that threatens the effectiveness of therapies for many infectious diseases. In this respect, antimicrobial peptides appear to be an interesting alternative to combat bacterial pathogens. Here, we report the characteristics of an antimicrobial peptide (of 30 amino acids) derived from the clostridial LysC enzyme. The peptide showed killing activity against clinical strains of Gram-positive and Gram-negative pathogens. Experimental data and computational modeling showed that this peptide forms transmembrane pores, directly engaging the negatively charged phospholipids of the bacterial cell membrane. Consequently, dissipation of the electrochemical gradient across cell membranes affects many vital processes, such as ATP synthesis, motility, and transport of nutrients. This kind of dysfunction leads to the loss of bacterial viability. Our firm conviction is that the presented study will be a helpful resource in searching for novel antimicrobial peptides that could have the potential to replace conventional antibiotics.

Difficulties in the Modeling of E. coli Spreading from Various Sources in a Coastal Marine Area.

Coastal and transitional waters are often used as bathing waters. In many regions, such activities play an important economic role. According to the European Union Bathing Water Directive (2006/7/EC) (BWD) the concentration of Escherichia coli in bathing water exceeding 500 CFU·100 mL-1 poses a high risk for bathers' health. In order to safeguard public health, microbiological environmental monitoring is carried out, which has recently been supported or replaced by mathematical models detailing the spread of sanitary contamination. This study focuses on the problems and limitations that can be encountered in the process of constructing a mathematical model describing the spread of biological contamination by E. coli bacteria in coastal seawater. This and other studies point to the following problems occurring during the process of building and validating a model: the lack of data on loads of sanitary contamination (often connected with multiple sources of biological contamination inflow) makes the model more complex; E. coli concentrations higher than 250 CFU·100 mL-1 (low hazard for health) are observed very rarely, and are associated with great uncertainty; the impossibility of predicting the time and intensity of precipitation as well as stronger winds and rougher sea, which may be a significant source of E. coli. However, there is universal agreement that such models will be useful in managing bathing water quality and protecting public health, especially during big failures of the wastewater network.

Raw Meat Contaminated with Cephalosporin-Resistant Enterobacterales as a Potential Source of Human Home Exposure to Multidrug-Resistant Bacteria.

The prevalence of cephalosporine-resistant (3GC-R) strains among United States community-related research samples ranged from 5.6 to 10.8%, while, in the European countries, it was 1.2% to 10.1%. Several studies suggest that meat of animal origin could be one of the reservoirs of 3GC-R bacteria. Here, 86 raw meat samples (turkey, pork, chicken and beef) were collected randomly and verified for the presence of 3GC-R bacteria. The 3GC-R bacteria were isolated, identified and characterized phenotypically (antibiotic resistance, motility and biofilm) and genotypically (repetitive-sequence-based rep-PCR) to elucidate any correlations with principal component analysis (PCA). From 28 3GC-R positive samples, 41 strains were isolated, from which the majority belonged to Serratia fonticola (39%), followed by Escherichia coli (19.5%), Enterobacter cloacae (17.1%) and Klebsiella pneumoniae (14.6%). The isolates of E. coli and S. fonticola presented diverse profiles in rep-PCR. Generally, 3GC-R strains were more resistant to antibiotics used in veterinary medicine than in human medicine. PCA derived from antibiotic resistance, motility and biofilm formation of S. fonticola and E. coli strains showed that resistance to beta-lactams was separated from the resistance to other antibiotic classes. Moreover, for the S. fonticola, E. coli and En. cloacae, the type of meat can create a specific tendency towards antibiotic resistance and phenotypic characteristics for S. fonticola, while these relationships were not found for other tested species.

Antibiotic resistance, virulence, and phylogenetic analysis of Escherichia coli strains isolated from free-living birds in human habitats.

Wild birds can be colonized by bacteria, which are often resistant to antibiotics and have various virulence profiles. The aim of this study was to analyze antibiotic resistance mechanisms and virulence profiles in relation to the phylogenetic group of E. coli strains that were isolated from the GI tract of wildfowl. Out of 241 faecal samples, presence of E. coli resistant to a cephalosporin (ESBL/AmpC) was estimated for 33 isolates (13,7%). Based on the analysis of the coexistence of 4 genes encoding ESBLs/AmpC (blaCTX-M, blaTEM, blaSHV, blaAmpC) and class 1 and 2 integrons genes (intI1, intI2) a subset of two resistance profiles was observed among the investigated E. coli isolates carrying blaAmpC, blaSHV, and blaCTX-M, blaTEM, class 1 and 2 integrons, respectively. The E. coli isolates were categorized into 4 phylogenetic groups A (39.4%), B2 (24.25%), D (24.25%) and B1 (12.1%). The pathogenic B2 and D groups were mainly typical for the Laridae family. Among the 28 virulence factors (Vfs) detected in pathogenic phylogenetic groups B2 and D, 7 were exclusively found in those groups (sfa, vat, tosA, tosB, hly, usp, cnf), while 4 VFs (fecA, fyuA, irp2, kspMTII) showed a statistically significant association (P≤0.05) with phylogroups A and B1. Our results indicated that strains belonging to commensal phylogroups A/B1 possess extensive iron acquisition systems (93,9%) and autotransporters (60,6%), typical for pathogens, hence we suggest that these strains evolve towards higher levels of virulence. This study, which is a point assessment of the virulence and drug resistance potential of wild birds, confirms the importance of taking wild birds as a reservoir of strains that pose a growing threat to humans. The E. coli analyzed in our study derive from different phylogenetic groups and possess an arsenal of antibiotic resistance genes and virulence factors that contribute to their ability to cause diseases.

Escherichia coli and Serratia fonticola ESBLs as a potential source of antibiotics resistance dissemination in the Tricity water reservoirs.

Despite the fact that cephalosporins are rarely used in medical or veterinary treatment, the presence of Enterobacterales strains resistant to this group of anti-bacterial drugs (ESBL) is an important issue that requires attention. Between 2019 and 2021, 14 retention reservoirs, 12 streams, 3 rivers and 1 lake situated in the Tricity area (in northern Poland) were sampled for the presence of ESBL strains. Out of 40 water samples, characteristic growth (E. coli and the KESC group) on Chromagar ESBL plates was observed for 33 samples. The average number of ESBL E. coli was 42±132 CFU/100 ml, while the KESC group was 73±147 CFU/100 ml. Out of 33 positive samples, 57 ESBL Enterobacterales strains were isolated, of which the most abundant species were E. coli (13 isolates) and S. fonticola (23 isolates). The E. coli ESBL isolates not only showed resistance to third generation cephalosporins but also to antibiotics from other groups, such as fluoroquinolones, aminoglycosides and sulfonamides. The S. fonticola ESBL isolates were also found to be mainly resistant to the third generation cephalosporins, with the exception of 5 imipenem and 2 ertapenem-resistant strains. These strains presented highly diverse fingerprinting profiles, as well as significant differences in phenotypic traits helpful for survival in the environment, such as biofilm formation and motility. Moreover, biofilm formation and the swimming ability were species and temperature dependent. We confirmed the presence of highly diverse ESBL strains with multiple drug resistance patterns in the Tricity water reservoirs. This could possibly pose a threat to human health and create a suitable ground for acquiring antibiotics resistance in the natural environment.

Multiresistant strains are as susceptible to photodynamic inactivation as their naïve counterparts: protoporphyrin IX-mediated photoinactivation reveals differences between methicillin-resistant and methicillin-sensitive Staphylococcus aureus strains.

OBJECTIVE: The current study was aimed at the investigation of differences in response to photoinactivation between methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive S. aureus (MSSA) isolates. Moreover, we aimed to elucidate if the observed variation resulted from antimicrobial resistance mechanisms and strains' susceptibility to antibiotic therapy. BACKGROUND DATA: Because of the emergence of multidrug resistance, the development of alternative antimicrobial strategies seems to be required. The concept of photodynamic inactivation (PDI) involves cell exposure to appropriate wavelength light that leads to the excitation of photosensitizer molecules, resulting in the production of reactive oxygen species responsible for cell inactivation and death. Recently, we have demonstrated a strain-dependent response of S. aureus to photoinactivation, and observed elevated resistance to PDI among MRSA strains. Nevertheless, the mechanism underlying this phenomenon remains unexplained. METHODS: S. aureus response to protoporphyrin IX (PPIX)-mediated photoinactivation was studied for 424 MRSA/MSSA isolates. VITEK 2 Advanced Expert System was used to detect antimicrobial resistance mechanisms and strains' susceptibility to antibiotictherapy. RESULTS: Data obtained demonstrated that MRSA are significantly more resistant to photoinactivation than MSSA strains; however, the difference observed did not result from antimicrobial susceptibility or resistance mechanisms. Furthermore, regardless of the strains' origin, a similar effectiveness of PDI could be achieved. Moreover, it was determined that the ability to form biofilms in vitro, and the presence of mec element, does not explain the observed differences between MRSA and MSSA strains. CONCLUSIONS: PDI could be highly effective against multidrug resistant pathogens as well as their naïve counterparts. Nevertheless, regardless of the antimicrobial resistance mechanism, the difference in response to PDI between MRSA and MSSA exists.

Mucormycosis complicating lower limb crash injury in a multiple traumatised patient: an unusual case.

Necrotising skin and soft tissues infections are most commonly bacterial in origin. However, saprophytic fungi of the class Zygomycetes, family Mucoraceae, can cause highly aggressive infections (mucormycoses) mainly in immunocompromised patients. Severe trauma is one of the major risk factors for mucormycosis. Fungal traumatic wound infection is an unusual complication associated with crash limb injury. This report describes a case of serious necrotising soft tissue infection caused by Mucor sp following primary fungal environmental wound contamination in a multiply injured patient. Despite undelayed diagnosis and proper treatment (surgical debridement and limb amputation, amphotericin B therapy) the patient presented a fatal outcome.

[Escherichia coli a forgotten pathogen in septicemia]. / Escherichia coli zapomniany patogen posocznic.

The aim of the retrospect study was to analyse the incidence of E. coli bacteremia in eight wards of SPSK 1 ACK AM in Gdansk from 2002 to 2004. We analyzed the incidence of bacteremia, patients outcome, source of infection and antimicrobial susceptibility. During the study period we detected 268 patients with E. coli bacteremia (8,0% of all bacteremic patients). 11,2% of them died within 24-48h after positive blood culture. Incidence of bacteremia was 1,7/1000 patients and the highest level achieved in Hematology Unit--33,2. The main portal of entry was genitourinary tract (24,3%) and gastrointestinal tract (21,8%). The strains (n=263) were least susceptible to ampicillin (33,3%), co-trimoxazole (68,4%), amoxycillin with clavulanic acid (69,3%) and ciprofloxacine (78,9%).

[Nosocomial bacteremia]. / Bakteriemie szpitalne.

Nosocomial bacteremia according to different authors can be divided in to bacteremia persistent, polymicrobial bacteremia, alternate bacteremia, breakthrough bacteremia, different bacteremia. Developing techniques of microbiological and molecular diagnostics induce us to verify the definition and partition of term "bacteremia".

[Study on genetic diversity of genes encoding main adhesins among nontypeable Haemophilus influenzae strains isolated from children in Poland]. / Badania profilu determinant genetycznych kodujacych glówne adhezyny bezotoczkowych szczepów Haemophilus influenzae izolowanych od dzieci w Polsce.

The study was based on hypothesis that in the nontypeable population of H. influenzae strains isolated from children there are some genetically predisposed to induce symptomatic infection in children and that they might be divided into different groups depending on profiles of genes encoding main adhesins synthesis. The work aimed at analysis of distribution of genes encoding adhesins and evaluation of domination possibility of some strains representing particular adhesins genes profiles among NTHi population. Results of the study revealed that among population of NTHi strains, distribution of genes encoding main adhesins are differing. Among children, NTHi strains harbouring genes encoding HA and HMW1/HMW2 adhesins were more prevalent in healthy children and in children with symptomatic infections, respectively. Analysis of strains harbouring main adhesins profiles might be a useful screening method in monitoring strains circulating among children, in order to determine the most invasive NTHi strains.

[Microorganisms analysis in Public Clinical Hospital in Gdansk over three years 2001-2003]. / Analiza wyników diagnostycznych badan mikrobiologicznych u pacjentów Szpitala Klinicznego w Gdansku, w latach 2001-2003.

The aim of the study was to analyse the changes in occurence of microorganisms and antibiotic usage in tertiary care hospital over 3 years. We analysed the results of microbiological records from laboratory inforation systems from 2001 to 2003. Over the study period there was about 40% increase of specimens received in the laboratory mainly due to another hospital incorporation. The relations between different groups of microorganisms was stable, Gram negatives 44,4%-46,3%, Gram positives 37,3%-40,3%, yeasts 7,0%-8,1%. There was a decrease in MRSA from 0,6% to 0,2% and carbapenem resistant Pseudomonas aeruginosa (CRPA) isolations from 2,0% to 0,7%, however the reverse was true for VRE, increase from 0,3% to 2%. ESBL-producing bacteria were isolated from about 4% of Enterobacteriaceae throughout the study. The analysis of blood cultures revealed over 60% deacrease in P. aeruginosa bacteremia and stable incidence of Escherichia coli (7%) and Staphylococcus aureus (6,5%) bacteremia. Increased usage of cephalosporins and fluoroquinolones was accompanied by the decrease in carbapenems and penicillins. In most cases there were no significant changes in occurence of main groups of microorganisms. Some multidrug resistant bacteria like MRSA and CRPA are no longer a problem in our hospital. Others like VRE, ESBL and Acinetobacter still cause concern due to high colonisation or infection rate. The usage of some antibiotic groups increased, another decreased and finally some like aminoglicosides and glicopeptides remained stable.

Molecular epidemiology of Serratia marcescens in two hospitals in Gdansk, Poland, over a 5-year period.

The history of the Serratia marcescens population in two hospitals in Danzig, Poland, over a 5-year period was analyzed in a study that combined MIC evaluation, typing by randomly amplified polymorphic DNA (RAPD) analysis and pulsed-field gel electrophoresis, and analysis of extended-spectrum beta-lactamases (ESBLs). We analyzed 354 isolates collected from 341 patients in two teaching hospitals in Danzig, Poland, from 1996 to 2000. The antimicrobial susceptibility profiles varied greatly, and for resistance to newer beta-lactams, probable AmpC cephalosporinase derepression and ESBL production occurred in about 23 and 19% of the isolates, respectively. RAPD typing, by which 69 types were discerned altogether, revealed a high degree of clonal diversity among the populations. However, the four most prevalent types were highly predominant, grouping approximately 71% of the isolates studied. These clones were observed in the two hospitals and were strong contributors to both outbreaks and the background of endemicity of the S. marcescens infections. Some of the strains that were not so widely spread (12 RAPD types; approximately 14% of the isolates) were responsible for several smaller outbreaks, and the remaining isolates represented unique RAPD types (53 types; approximately 15% of the isolates) and were probably sporadic introductions from other environments. ESBLs were identified in several different clones, and some of these had most likely already been introduced into the hospitals as ESBL producers, whereas the others acquired the ESBL-encoding genes from other enterobacterial strains in these environments. The CTX-M-3 enzyme, which is widely observed in Poland, was the most common ESBL type among the S. marcescens isolates, followed by TEM-47 and SHV-5. The complex epidemiology of ESBLs, especially in 1999 and 2000, must have arisen from the introduction of ESBL producers from other centers, their clonal dissemination, and the constant penetration of the S. marcescens populations with plasmids with ESBL genes. Multiple S. marcescens isolates were obtained from 11 patients, who probably represented both patients with recolonizations and reinfections and patients with recurrences of infections with the evolution of the strain's susceptibility.

[Microorganisms isolated from clinical specimens of patients from the Department of Thoracic Surgery]. / Drobnoustroje izolowane z materialów klinicznych torakochirurgicznej chorych z kliniki.

The clinical specimens received from patients hospitalized in Department of Thoracic Surgery between 1997 and 2001 were microbiologically examined. The main specimen for microbiological examination was pleural fluid (median 34%). The frequency of specimens from bronchial tree increased significantly (from 4% to 26%) with concurrent decrease of sputum (from 29% to 6%). Among isolated pathogens, Gram negative rods were the most frequent (median 48%) and Pseudomonas sp. was the main pathogen among them. Occurrence of staphylococci was median 22% and Staphylococcus aureus, with a little decrease in analyzed period, was still the main Gram positive pathogen. Simultaneously the occurrence of MRSA in the last three years dropped three times. The number of isolations of yeasts have risen from 5.8% to 10.3%.