Large hospital outbreak caused by OXA-244-producing Escherichia coli sequence type 38, Poland, 2023.

In February 2023, Escherichia coli sequence type (ST) 38 producing oxacillinase 244 (OXA-244-Ec ST38) was detected from three patients in a hospital in western Poland. Overall, OXA-244-Ec ST38 was detected from 38 colonised patients in 13 wards between February and June 2023. The outbreak was investigated on site by an infection control team, and the bacterial isolates were characterised microbiologically and by whole genome sequencing. We could not identify the primary source of the outbreak or reconstruct the transmission sequence. In some of the 13 affected wards or their groups linked by the patients' movement, local outbreaks occurred. The tested outbreak isolates were resistant to ß-lactams (penicillins, cephalosporins, aztreonam and ertapenem) and to trimethoprim-sulfamethoxazole. Consistently, apart from bla OXA-244, all isolates contained also the bla CMY-2 and bla CTX-M-14 genes, coding for an AmpC-like cephalosporinase and extended-spectrum ß-lactamase, respectively, and genes conferring resistance to trimethoprim-sulfamethoxazole, sul2 and dfrA1. Genomes of the isolates formed a tight cluster, not of the major recent European Cluster A but of the older Cluster B, with related isolates identified in Germany. This outbreak clearly demonstrates that OXA-244-Ec ST38 has a potential to cause hospital outbreaks which are difficult to detect, investigate and control.

Dissemination of extensively drug-resistant NDM-producing Providencia stuartii in Europe linked to patients transferred from Ukraine, March 2022 to March 2023.

BackgroundThe war in Ukraine led to migration of Ukrainian people. Early 2022, several European national surveillance systems detected multidrug-resistant (MDR) bacteria related to Ukrainian patients.AimTo investigate the genomic epidemiology of New Delhi metallo-ß-lactamase (NDM)-producing Providencia stuartii from Ukrainian patients among European countries.MethodsWhole-genome sequencing of 66 isolates sampled in 2022-2023 in 10 European countries enabled whole-genome multilocus sequence typing (wgMLST), identification of resistance genes, replicons, and plasmid reconstructions. Five bla NDM-1-carrying-P. stuartii isolates underwent antimicrobial susceptibility testing (AST). Transferability to Escherichia coli of a bla NDM-1-carrying plasmid from a patient strain was assessed. Epidemiological characteristics of patients with NDM-producing P. stuartii were gathered by questionnaire.ResultswgMLST of the 66 isolates revealed two genetic clusters unrelated to Ukraine and three linked to Ukrainian patients. Of these three, two comprised bla NDM-1-carrying-P. stuartii and the third bla NDM-5-carrying-P. stuartii. The bla NDM-1 clusters (PstCluster-001, n = 22 isolates; PstCluster-002, n = 8 isolates) comprised strains from seven and four countries, respectively. The bla NDM-5 cluster (PstCluster-003) included 13 isolates from six countries. PstCluster-001 and PstCluster-002 isolates carried an MDR plasmid harbouring bla NDM-1, bla OXA-10, bla CMY-16, rmtC and armA, which was transferrable in vitro and, for some Ukrainian patients, shared by other Enterobacterales. AST revealed PstCluster-001 isolates to be extensively drug-resistant (XDR), but susceptible to cefiderocol and aztreonam-avibactam. Patients with data on age (n = 41) were 19-74 years old; of 49 with information on sex, 38 were male.ConclusionXDR P. stuartii were introduced into European countries, requiring increased awareness and precautions when treating patients from conflict-affected areas.

Genome-Based Epidemiologic Analysis of VIM/IMP Carbapenemase-Producing Enterobacter spp., Poland.

We sequenced all nonduplicate 934 VIM/IMP carbapenemase-producing Enterobacterales (CPE) reported in Poland during 2006-2019 and found ≈40% of the isolates (n = 375) were Enterobacter spp. During the study period, incidence of those bacteria gradually grew in nearly the entire country. The major factor affecting the increase was clonal spread of several E. hormaechei lineages responsible for multiregional and interregional outbreaks (≈64% of all isolates), representing mainly the pandemic sequence type (ST) 90 or the internationally rare ST89 and ST121 clones. Three main VIM-encoding integron types efficiently disseminated across the clone variants (subclones) with various molecular platforms. Those variants were predominantly Pseudomonas aeruginosa-derived In238-like elements, present with IncHI2+HI2A, IncFII+FIA, IncFIB, or IncN3 plasmids, or chromosomal genomic islands in 30 Enterobacter STs. Another prevalent type, found in 34 STs, were In916-like elements, spreading in Europe recently with a lineage of IncA-like plasmids.

Epidemic Territorial Spread of IncP-2-Type VIM-2 Carbapenemase-Encoding Megaplasmids in Nosocomial Pseudomonas aeruginosa Populations.

In 2003 to 2004, the first five VIM-2 metallo-ß-lactamase (MBL)-producing Pseudomonas aeruginosa (MPPA) isolates with an In4-like integron, In461 (aadB-blaVIM-2-aadA6), on conjugative plasmids were identified in three hospitals in Poland. In 2005 to 2015, MPPA expanded much in the country, and as many as 80 isolates in a collection of 454 MPPA (∼18%) had In461, one of the two most common MBL-encoding integrons. The organisms occurred in 49 hospitals in 33 cities of 11/16 main administrative regions. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) classified them into 55 pulsotypes and 35 sequence types (STs), respectively, revealing their remarkable genetic diversity overall, with only a few small clonal clusters. S1 nuclease/hybridization assays and mating of 63 representative isolates showed that ∼85% of these had large In461-carrying plasmids, ∼350 to 550 kb, usually self-transmitting with high efficiency (∼10-1 to 10-2 per donor cell). The plasmids from 19 isolates were sequenced and subjected to structural and single-nucleotide-polymorphism (SNP)-based phylogenetic analysis. These formed a subgroup within a family of IncP-2-type megaplasmids, observed worldwide in pseudomonads from various environments and conferring resistance/tolerance to multiple stress factors, including antibiotics. Their microdiversity in Poland arose mainly from acquisition of different accessory fragments, as well as new resistance genes and multiplication of these. Short-read sequence and/or PCR mapping confirmed the In461-carrying plasmids in the remaining isolates to be the IncP-2 types. The study demonstrated a large-scale epidemic spread of multidrug resistance plasmids in P. aeruginosa populations, creating an epidemiological threat. It contributes to the knowledge on IncP-2 types, which are interesting research objects in resistance epidemiology, environmental microbiology, and biotechnology.

Citrobacter telavivum sp. nov. with chromosomal mcr-9 from hospitalized patients.

Strains 6105T and 6106, recovered from colonized patients in a hospital in Tel-Aviv, Israel, were compared with currently known species of the genus Citrobacter by a polyphasic taxonomic approach. Strains were characterized by whole-genome sequencing, 16S rRNA and recN gene sequencing, multilocus sequence analysis (MLSA), average nucleotide identity (ANI), Genome-to-Genome Distance Calculator (GGDC), and biochemical tests. The location and genetic surrounding of antibiotic resistance genes were investigated, and antibiotic susceptibility profiles were determined by broth microdilution or agar dilution methods. Phylogenetic analysis based on recN and MLSA revealed that both strains formed a distinct cluster from all currently recognized species. The ANI and GGDC were 90.7% and 54.3% with Citrobacter farmeri, respectively. The ability to metabolize various compounds also differentiated both strains from closely related Citrobacter species. Chromosomes of the isolates contained locus encoding a novel class A ß-lactamase (TEL-1; 90.5% amino acid identity with CdiA of Citrobacter koseri) plus a LysR-like transcriptional regulator (TEL-R) and an ~ 25.5-kb mcr-9 mosaic region. The direct mcr-9 context matched with those previously identified in several plasmids and chromosomes of diverse Enterobacteriaceae, yet similarity with the plasmidic loci extended further. Untypeable plasmids, pCTEL-2 (~ 235 kb) and pCTEL-1 (~ 114 kb), devoid of resistance genes, were identified in the strains. The isolates were non-susceptible to ß-lactams. The name Citrobacter telavivum sp. nov. is proposed, with 6105T (CECT 9989T or DSM 110286T) as the type strain. C. telavivum may represent a bacterial species adapting to hospital settings, able to disseminate and acquire antimicrobial resistance genes.

The Impact of Insertion Sequences on O-Serotype Phenotype and Its O-Locus-Based Prediction in Klebsiella pneumoniae O2 and O1.

Klebsiella pneumoniae is a nosocomial pathogen, pointed out by the World Helth Organisation (WHO) as "critical" regarding the highly limited options of treatment. Lipopolysaccharide (LPS, O-antigen) and capsular polysaccharide (K-antigen) are its virulence factors and surface antigens, determining O- and K-serotypes and encoded by O- or K-loci. They are promising targets for antibody-based therapies (vaccines and passive immunization) as an alternative to antibiotics. To make such immunotherapy effective, knowledge about O/K-antigen structures, drift, and distribution among clinical isolates is needed. At present, the structural analysis of O-antigens is efficiently supported by bioinformatics. O- and K-loci-based genotyping by polymerase chain reaction (PCR) or whole genome sequencing WGS has been proposed as a diagnostic tool, including the Kaptive tool available in the public domain. We analyzed discrepancies for O2 serotyping between Kaptive-based predictions (O2 variant 2 serotype) and the actual phenotype (O2 variant 1) for two K. pneumoniae clinical isolates. Identified length discrepancies from the reference O-locus resulted from insertion sequences (ISs) within rfb regions of the O-loci. In silico analysis of 8130 O1 and O2 genomes available in public databases indicated a broader distribution of ISs in rfbs that may influence the actual O-antigen structure. Our results show that current high-throughput genotyping algorithms need to be further refined to consider the effects of ISs on the LPS O-serotype.

Review of measles-related events recorded by the National IHR Focal Point in Poland in the years 2016-2018

Member States of the World Health Organization (WHO), in accordance with the requirements of the International Health Regulations (2005), were obliged to establish National Focal Points for International Health Regulations (IHR NFP), whose task is, among others, consolidating information on public health events of international importance that occur abroad or in the country. The aim of this article is to review information on measles-related events posted on the Event Information Site for IHR National Focal Points, in the Early Warning and Response System (EWRS), received by email directly from other IHR National Focal Points located in WHO member states, and from all organs of the State Sanitary Inspectorate in Poland in the years 2016-2018. In this time period, the IHR NFP recorded 92 measles-related events of which 38 related to individual cases, 37 to outbreaks of the disease, and 17 involved exposure to a measles case. 36% of reported events were aviationrelated. The number of events in 2018 has tripled compared to 2017 and increased eightfold in comparison to 2016. The current situation indicates the need to take appropriate actions, including implementation of the National Vaccination Program as well as introducing vaccination interventions.

Multiregional dissemination of KPC-producing Klebsiella pneumoniae ST258/ST512 genotypes in Poland, 2010-14.

Objectives: In 2008-09, the KPC carbapenemase epidemiology in Poland was dominated by a Klebsiella pneumoniae ST258 KPC-2 outbreak in Warsaw and its administrative region. The aim of this study was to analyse the situation in 2010-14, with a focus on new outbreaks in other parts of the country. Methods: KPCs were detected in all suspected isolates by PCR. The detailed study was performed on 173 isolates from 2010 to 2012, and included PFGE and MLST, PCR identification of K. pneumoniae clonal group CG258 clades and potential specificity markers ( pilv-1 , IS 66 and prp ), PCR mapping of Tn 4401 transposons, and plasmid analysis by nuclease S1 profiling and PCR-based replicon typing. Results: Six hundred and eight KPC cases were identified in Poland in 2010-14, almost half of which occurred in the Warsaw region, and another half in four other areas. The new outbreaks were caused by four K. pneumoniae CG258 genotypes, different from each other and from the organisms spreading in Warsaw. The new lineages were ST258 or ST512 of clade II, and had specific compositions of potential ST258/ST512 clonal markers. The isolates produced KPC-3 encoded by Tn 4401 a or Tn 4401 b elements on plasmids with single or multiple replicons, including I2, FII K (+/-FIB K ), 'FII Y -like', X3 and R. Of other species, Citrobacter freundii ST17 and Enterobacter cloacae ST254 with KPC-2 were identified in a Warsaw hospital. Conclusions: The study showed remarkable changes in the KPC epidemiology in Poland in 2010-14, which, following the localized regional spread in the early phase, has converted into multiregional dissemination.

Measles elimination ­ review of event notifications sent to National IHR Focal Point between 2010 and 2016

The Member States of the World Health Organization (WHO) in accordance with International Health Regulations (2005) were obliged to appoint National IHR Focal Points (N IHR FP), of which tasks include obtaining information concerning public health emergencies of international concern which occurred abroad or within the country. The aim of this work is the review of WHO, ECDC, National IHR Focal Points from the WHO Member States and The State Sanitary Inspection notifications related to measles received by National IHR Focal Point in Poland in the period from 2010 to 2016. During this period N IHR FP was informed about 79 events related to measles. These events include: 36 related to the outbreaks in different countries, 27 concerning individual cases, 14 related to the exposure in contact with a measles case during air travel and two concerning the implementation of the MMR vaccination programs. Despite the progress in implementing the measures included in the elimination of measles programs in Europe, there was a significant increase in the number of measles cases and outbreaks particularly in years 2010-2011.

Poliomyelitis eradication ­ the review of notifications from the years 2010-2016 sent to National IHR Focal Point

Polio eradication programme was launched after World Health Assembly in 1988. Despite considerable decrease in reported cases it still constitutes a significant public health threat. All WHO member state is bound to appoint National IHR Focal Point, which operates based on International Health Regulations (2005), which were enacted during the World Health Assembly in 2005. In Poland National IHR Focal Point (IHR NFP in Poland) operates since 2007, and is located in the Department of Epidemiology, in National Institute of Public Health - National Institute of Hygiene. Its aim is to acquire, assess and to transfer information on events which may constitute an international threat for the public health. IHR NFP in Poland has an access to WHO's Event Information Site (EIS) as well as Early Warning and Response System (EWRS) with reading-only credentials. Both platforms are of limited access (1). Among recipients of IHR NFP notifications and information are experts from many fields such as epidemiology, virology, bacteriology and others- related to specific type of notification, as well as specific and appointed members of state's administration and authorities in the field of public health. In this paper a review of notifications on the subject of poliomyelitis, sent to IHR NFP in Poland in the years 2010-2016 is presented, as well as references to poliomyelitis epidemiological situation were made based on the date from Global Polio Eradication Initiative.

Assessment of knowledge about the effects of UV radiation on health behaviors associated with sunbathing in gymnasium students.

INTRODUCTION: The health behaviors or health-related behaviors is behavior (or activity) that are part of everyday life, affecting the health of the individual. An example of the behavior of health is also sunbathing, or exposing the body to excessive solar radiation dosage. It may be positive and negative effects on health. AIM: Evaluation of knowledge about gymnasium students. The impact of UV radiation on health and health behaviors associated with sunbathing. MATERIAL AND METHODS: The study was a diagnostic survey, with author's questionnaire. The sample was comprised students from classes II and III. A total of 312 questionnaires were collected among 181 girls and 131 boys. Used purposeful sampling. Results were considered statistically significant at p < 0.05. RESULTS: There is a statistically significant relationship between the stated sunbathing to get a sun tan, and sex of the respondent (p = 0.0002). Definitely more girls (77.35%) admit that tans in the sun to get a tan as compared to boys (58.02%). CONCLUSION: It is recommended that further research aimed at checking the causes and incidence of sunburn among young people. Consideration should be given to create and implement the appropriate health programs taking about tanning that could be implemented under the School Health Promotion Program, or to supplement this knowledge on subjects such as Biology or Nature.

Characterization of pKP-M1144, a Novel ColE1-Like Plasmid Encoding IMP-8, GES-5, and BEL-1 ß-Lactamases, from a Klebsiella pneumoniae Sequence Type 252 Isolate.

IMP-8 metallo-ß-lactamase was identified in Klebsiella pneumoniae sequence type 252 (ST252), isolated in a Portuguese hospital in 2009. blaIMP-8 was the first gene cassette of a novel class 3 integron, In1144, also carrying the blaGES-5, blaBEL-1, and aacA4 cassettes. In1144 was located on a ColE1-like plasmid, pKP-M1144 (12,029 bp), with a replication region of limited nucleotide similarity to those of other RNA-priming plasmids, such as pJHCMW1. In1144 and pKP-M1144 represent an interesting case of evolution of resistance determinants in Gram-negative bacteria.

Matrix-assisted laser desorption ionization-time of flight mass spectrometry meropenem hydrolysis assay with NH4HCO3, a reliable tool for direct detection of carbapenemase activity.

A comparison of a matrix-assisted laser desorption ionization-time of flight mass spectrometric (MALDI-TOF MS) meropenem hydrolysis assay with the Carba NP test showed that both methods exhibited low sensitivity (approximately 76%), mainly due to the false-negative results obtained with OXA-48-type producers. The addition of NH4HCO3 to the reaction buffer for the MALDI-TOF MS assay dramatically improved its sensitivity (98%). Automatic interpretation of the MALDI-TOF MS assay, using the MBT STAR-BL software, generally agreed with the results obtained after manual analysis. For the Carba NP test, spectrophotometric analysis found six additional carbapenemase producers.

PCR melting profile as a tool for outbreak studies of Salmonella enterica in chickens.

BACKGROUND: Salmonellosis is of great economic concern in all phases of the poultry industry, from production to marketing, leading to severe economic losses. Monitoring the source of the bacterial contamination has fundamental importance in the spreading of salmonellosis. RESULTS: We applied a ligation-mediated PCR method, PCR MP (PCR melting profile), to type S. enterica ssp. enterica ser. Enteritidis (56 strains) and 43 control strains classified to other serovars isolated from poultry. We demonstrated the PCR MP potential for salmonellosis spreading monitoring. Our rapid test presents higher discriminatory power (0.939 vs. 0.608) compared to current molecular subtyping tool such as pulsed-field gel electrophoresis (PFGE), which ineffectiveness underlies the high degree of clonality of S. Enteritidis. CONCLUSIONS: PCR MP was found to be a highly discriminating, sensitive and specific method that could be a valuable molecular tool, particularly for analyzing epidemiological links of limited number of S. enterica ser. Enteritidis strains.

Emerging infectious diseases not covered by routine vaccination in Europe in 2010-2015--the review of WHO and ECDC notifications for the National IHR Focal Point in Poland.

The National IHR Focal Point is a center set up by each Member State of the World Health Organization (WHO) in accordance with the International Health Regulations (2005). The International Health Regulations (IHR) were adopted on 23 May 2005 at the World Health Assembly and entered into force since 15 June 2007 as the legal instrument designed to help protect all countries from uncontrolled international spread of diseases and other urgent public health threats. According to Article 2 of IHR the purpose and scope of these Regulations are to prevent, protect against, control and provide a public health response to the international spread of disease in ways that are commensurate with and restricted to public health risks, and which avoid unnecessary interference with international traffic and trade. Primarily, the scope of IHR is to establish a system of early warning (in accordance with Article 6 and 7) with the functioning in each country National IHR Focal Point which is available at any time to communicate with WHO IHR Contact Points and other entities. The tasks of the National IHR Fo- cal Point in Poland which was appointed by the Minister of Health and runs in the Department of Epidemiology, National Institute of Public Health--National Institute of Hygiene from 1 September 2007 are the notification of events that may constitute a public health emergency of international concern occurring in Poland or abroad and the dissemination of this information to the WHO, other National IHR Focal Points or competent authorities responsible for public health. The task of the National IHR Focal Point in Poland is also the dissemination of WHO and ECDC notifications, including recommendation and risk assessment documents. The aim of this work is the review of WHO and ECDC notifications received by National IHR Focal Point in Poland in the period from 2010 to 2015 which were related to emerging infectious diseases not covered by routine vaccination programs or for which there are no effective vaccines that have occurred in the WHO European Region. The review includes verotoxin-producing Escherichia coli O104: H4 infections, MERS-CoV infections, Ebola virus disease, malaria, dengue fever, West Nile fever, chikungunya and cholera.

Sequence types 235, 111, and 132 predominate among multidrug-resistant pseudomonas aeruginosa clinical isolates in Croatia.

A population analysis of 103 multidrug-resistant Pseudomonas aeruginosa isolates from Croatian hospitals was performed. Twelve sequence types (STs) were identified, with a predominance of international clones ST235 (serotype O11 [41%]), ST111 (serotype O12 [15%]), and ST132 (serotype O6 [11%]). Overexpression of the natural AmpC cephalosporinase was common (42%), but only a few ST235 or ST111 isolates produced VIM-1 or VIM-2 metallo-ß-lactamases or PER-1 or GES-7 extended-spectrum ß-lactamases.

The OL101 O antigen locus specifies a novel Klebsiella pneumoniae serotype O13 structure.

Klebsiella pneumoniae is one of the priority objects for the development of new therapies against infections. The species has been perceived as of limited variety of O antigens (11 O serotypes identified to date). That trait makes lipopolysaccharide an attractive target for protective antibodies. Nowadays, K. pneumoniae O antigens encoding genes are often analysed by bioinformatic tools, such as Kaptive, indicating higher actual diversity of the O antigen loci. One of the novel K. pneumoniae O loci for which the antigen structure has not been elucidated so far is OL101. In this study, four clinical isolates predicted as OL101 were characterized and found to have the O antigen structure composed of ß-Kdop-[→3)-α-l-Rhap-(1→4)-α-d-Glcp-(1→]n, representing a novel serotype O13. Identification of the ß-Kdop terminus was based on the analysis of the complete LPS molecule by the HR-MAS NMR spectroscopy. The bioinformatic analysis of 71,377 K. pneumoniae genomes from public databases (July 2023) revealed a notable OL101 prevalence of 6.55 %.

Genomic Epidemiology of MBL-Producing Pseudomonas putida Group Isolates in Poland.

INTRODUCTION: Pseudomonas putida group are described as low-incidence opportunistic pathogens, but also as a significant reservoir of antimicrobial resistance (AMR) genes, including those of metallo-ß-lactamases (MBLs). Our objective was the molecular and genomic characterization of MBL-producing P. putida (MPPP) group isolates from Poland, focusing on population structures, successful genotypes and MBL-encoding integrons. METHODS: During a country-wide MBL surveillance in Pseudomonas spp., 59 non-duplicate MPPP isolates were collected from 36 hospitals in 23 towns from 2003 to 2016. All of the isolates were subjected to whole-genome sequencing (WGS), followed by species identification, multi-locus sequence typing (MLST), single-nucleotide polymorphism (SNP)-based phylogenetic/clonality analysis, resistome determination, and susceptibility testing. RESULTS: The study collection comprised 12 species, of which P. alloputida (n = 19), P. monteilii (n = 15), and P. asiatica (n = 11) prevailed, while the others were P. kurunegalensis, P. putida, P. soli, P. mosselii, P. juntendi, and four potentially new species. MLST classified the isolates into 23 sequence types (STs) of which 21 were new, with three main clones, namely P. alloputida ST69, P.monteilii ST95 and P. asiatica ST15. The isolates produced VIM-like MBLs only, largely VIM-2 (n = 40), encoded by 24 different class 1 integrons (ten new), a number of which occurred also in P. aeruginosa and/or Enterobacterales in Poland. The plasmid pool was dominated by IncP-9, IncP-2, and pMOS94-like types. Multiple isolates were extensively drug-resistant. CONCLUSIONS: This study, being one of the most comprehensive analyses of MPPP so far, has shown high diversity of the isolates in general, with three apparently international lineages, each internally diversified by MBL-encoding structures.

Molecular characteristics of KPC-producing Enterobacteriaceae at the early stage of their dissemination in Poland, 2008-2009.

After the first report in May 2008, the National Reference Center for Susceptibility Testing confirmed 113 cases of infection or colonization by KPC-producing members of the family Enterobacteriaceae in Poland by the end of 2009. The vast majority of patients were found in 18 hospitals; three patients were diagnosed at outpatient clinics. Most of the institutions were in the Warsaw area, including three hospitals with the highest numbers of cases. When available, the data on previous hospitalizations often indicated that these hospitals were the probable acquisition sites; one patient arrived from New York. The group of 119 unique isolates consisted of Klebsiella pneumoniae (n = 114), followed by Klebsiella oxytoca (n = 3), and Escherichia coli (n = 2). The K. pneumoniae isolates were dominated by the clone sequence type 258 (ST258) (n = 111); others were ST11 and ST23. The ST258 group was heterogeneous, with 28 pulsed-field gel electrophoresis (PFGE) subtypes, ∼25 plasmid profiles, and nine ß-lactamase patterns differing by KPC variants (KPC-2 mainly), and SHV-12, CTX-M-3, and TEM-1-like enzymes. Plasmids carrying bla(KPC) genes varied in size (~48 to 250 kb), structure, and conjugation potential. Transferable IncFII(K) plasmids of ~110 to 160 kb, probably pKpQIL or its derivatives, were observed in all K. pneumoniae clones and in K. oxytoca. Also prevalent were nontypeable pETKp50-like plasmids of ~50 kb, found in K. pneumoniae ST258 and E. coli isolates (ST93 and ST224). Two K. pneumoniae-E. coli pairs from single patients might represent the in vivo transfer of such plasmids. The striking diversity of KPC producers at the early stage of dissemination could result from several introductions of these bacteria into the country, their multidirectional evolution during clonal spread, and transfer of the plasmids.

A new rapid and cost-effective method for detection of phages, ICEs and virulence factors encoded by Streptococcus pyogenes.

Streptococcus pyogenes (group A Streptococcus, GAS) is a human pathogen that causes diseases of various intensity, from mild strep throat to life threatening invasive infections and postinfectional sequelae. S. pyogenes encodes multiple, often phage encoded, virulence factors and their presence is related to severity of the disease. Acquisition of mobile genetic elements, carrying virulence factors, as phages or ICEs (integrative and cojugative elements) has been shown previously to promote selection of virulent clones. We designed the system of eight low volume multi- and one singleplex PCR reactions to detect genes encoding twenty virulence factors (spd3, sdc, sdaB, sdaD, speB, spyCEP, scpA, mac, sic, speL, K, M, C, I, A, H, G, J, smeZ and ssa) and twenty one phage and ICE integration sites described so far for S. pyogenes. Classification of strains based on the phage and virulence factors absence or presence, correlates with PFGE MLST and emm typing results. We developed a novel, fast and cost effective system that can be used to detect GAS virulence factors. Moreover, this system may become an alternative and effective system to differentiate between GAS strains.