The aim of this study was to characterize the first cases and outbreaks of OXA-48-like-producing Enterobacteriaceae recovered from hospital settings in the Czech Republic. From 2013 to 2015, 22 Klebsiella pneumoniae isolates, 3 Escherichia coli isolates, and 1 Enterobacter cloacae isolate producing OXA-48-like carbapenemases were isolated from 20 patients. Four of the patients were colonized or infected by two or three different OXA-48-like producers. The K. pneumoniae isolates were classified into nine sequence types (STs), with ST101 being predominant (n ϭ 8). The E. coli isolates were of different STs, while the E. cloacae isolate belonged to ST109. Twenty-four isolates carried bla OXA-48 , while two isolates carried bla OXA-181 or bla . Almost all isolates (n ϭ 22) carried bla OXA-48 -positive plasmids of a similar size (ϳ60 kb), except the two isolates producing OXA-181 or OXA-232. In an ST45 K. pneumoniae isolate and an ST38 E. coli isolate, S1 nuclease profiling plus hybridization indicated a chromosomal location of bla . Sequencing showed that the majority of bla OXA-48 -carrying plasmids exhibited high degrees of identity with the pOXA-48-like plasmid pE71T. Additionally, two novel pE71T derivatives, pOXA-48_30715 and pOXA-48_30891, were observed. The bla OXA-181 -carrying plasmid was identical to the IncX3 plasmid pOXA181_EC14828, while the bla OXA-232 -carrying plasmid was a ColE2-type plasmid, being a novel derivative of pOXA-232. Finally, sequencing data showed that the ST45 K. pneumoniae and ST38 E. coli isolates harbored the IS1R-based composite transposon Tn6237 containing bla OXA-48 integrated into their chromosomes. These findings underlined that the horizontal transfer of pOXA-48-like plasmids has played a major role in the dissemination of bla in the Czech Republic. In combination with the difficulties with their detection, OXA-48 producers constitute an important public threat.

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The objective of this study was to perform molecular surveillance for assessing the spread of carbapenemase-producing in Czech hospitals. One hundred thirty-six carbapenemase-producing isolates were recovered from 22 hospitals located throughout the country. Sequence type 357 (ST357) dominated ( = 120) among carbapenemase producers. One hundred seventeen isolates produced IMP-type (IMP-7 [ = 116] and IMP-1 [ = 1]) metallo-β-lactamases (MβLs), 15 produced the VIM-2 MβL, and the remaining isolates expressed the GES-5 enzyme. The -like genes were located in three main integron types, with In-p110-like being the most prevalent ( = 115). The two other IMP-encoding integrons (In1392 and In1393) have not been described previously. -carrying integrons included In59-like, In56, and a novel element (In1391). was carried by In717. Sequencing data showed that In-p110-like was associated with a Tn-like transposon inserted in genomic island LESGI-3 in the chromosome. The other integrons were also integrated into the chromosome. These findings indicated the clonal spread of ST357 , carrying the IMP-7-encoding integron In-p110, in Czech hospitals. Additionally, the sporadic emergence of producing different carbapenemase types, associated with divergent or novel integrons, punctuated the ongoing evolution of these bacteria.

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The aim of the present study was to characterize sporadic cases and an outbreak of NDM-like-producing Enterobacteriaceae recovered from hospital settings, in Czechia. During 2016, 18 Entrobacteriaceae isolates including 10 Enterobacter cloacae complex (9 E. xiangfangensis and 1 E. asburiae), 4 Escherichia coli, 1 Kluyvera intermedia, 1 Klebsiella pneumoniae, 1 Klebsiella oxytoca, and 1 Raoultella ornithinolytica that produced NDM-like carbapenemases were isolated from 15 patients. Three of the patients were colonized or infected by two different NDM-like producers. Moreover, an NDM-4-producing isolate of E. cloacae complex, isolated in 2012, was studied for comparative purposes. All isolates of E. cloacae complex, except the E. asburiae, recovered from the same hospital, were assigned to ST182. Additionally, two E. coli belonged to ST167, while the remaining isolates were not clonally related. Thirteen isolates carried blaNDM−4, while six isolates carried blaNDM−1 (n = 3) or blaNDM−5 (n = 3). Almost all isolates carried blaNDM-like-carrying plasmids being positive for the IncX3 allele, except ST58 E. coli and ST14 K. pneumoniae isolates producing NDM-1. Analysis of plasmid sequences revealed that all IncX3 blaNDM-like-carrying plasmids exhibited a high similarity to each other and to previously described plasmids, like pNDM-QD28, reported from worldwide. However, NDM-4-encoding plasmids differed from other IncX3 plasmids by the insertion of a Tn3-like transposon. On the other hand, the ST58 E. coli and ST14 K. pneumoniae isolates carried two novel NDM-1-encoding plasmids, pKpn-35963cz, and pEsco-36073cz. Plasmid pKpn-35963cz that was an IncFIB(K) molecule contained an acquired sequence, encoding NDM-1 metallo-β-lactamase (MβL), which exhibited high similarity to the mosaic region of pS-3002cz from an ST11 K. pneumoniae from Czechia. Finally, pEsco-36073cz was a multireplicon A/C2+R NDM-1-encoding plasmid. Similar to other type 1 A/C2 plasmids, the blaNDM−1 gene was located within the ARI-A resistance island. These findings underlined that IncX3 plasmids have played a major role in the dissemination of blaNDM-like genes in Czech hospitals. In combination with further evolvement of NDM-like-encoding MDR plasmids through reshuffling, NDM-like producers pose an important public threat.

sequence type 131 (ST131) is currently one of the leading causes of multidrug-resistant extraintestinal infections globally. Here, we analyzed the phenotypic and genotypic characteristics of 169 ST131 isolates from various sources (wildlife, wastewater, companion animals, community, and hospitals) to determine whether wildlife and the environment share similar strains with humans, supporting transmission of ST131 between different ecological niches. Susceptibility to 32 antimicrobials was tested by disc diffusion and broth microdilution. Antibiotic resistance genes, integrons, plasmid replicons, 52 virulence genes, and -based subtypes were detected by PCR and DNA sequencing. Genomic relatedness was determined by pulsed-field gel electrophoresis (PFGE). The genetic context and plasmid versus chromosomal location of extended-spectrum beta-lactamase and AmpC beta-lactamase genes was determined by PCR and probe hybridization, respectively. The 169 ST131 study isolates segregated predominantly into30Rx (60%) and 30R1 (25%) subclones. Within each subclone, isolates from different source groups were categorized into distinct PFGE clusters; genotypic characteristics were fairly well conserved within each major PFGE cluster. Irrespective of source, the30Rx isolates typically exhibited virotype A (89%), an F2:A1:B- replicon (84%), and a 1.7-kb class 1 integron (92%) and had diverse structures upstream of the region. In contrast, the30R1 isolates typically exhibited virotype C (86%), an F1:A2:B20 replicon (76%), and a conserved IS-ΔIS-like structure. Despite considerable overall genetic diversity, our data demonstrate significant commonality between ST131 isolates from diverse environments, supporting transmission between different sources, including humans, environment, and wildlife.

Abstract. We report a case of congenital fibropapillomatosis in the head and neck of a piglet from a swine breeding farm where sporadic cutaneous papillomas of the prepuce and scrotum had previously occurred in several boars. Histologically, the tumor consisted of multiple exophytic, papillary projections composed of branching, densely cellular, fibrovascular stalks covered by hyperplastic keratizing squamous epithelium. Ultrastructural examination failed to reveal viral particles. Cutaneous papillomatosis is rare in swine. To the best of our knowledge, this is the second report of congenital fibropapillomatosis in pigs.Key words: Congenital papillomatosis, fibropapillomatosis, head and neck, piglet, skin, ultrastructure. Papillomatosis is a self-limiting transmissible disease that in most cases has been shown to have a viral cause. The disease may occur in many domestic and wild animal species, and it is most common in cattle and horses. 3 Papillomatosis is rare in pigs, usually affecting the skin and the genitalia. 3 To the best of our knowledge, there is only one case of congenital papillomatosis in the pig previously reported in the literature. 4 In this report, we describe a case of congenital cutaneous fibropapillomatosis in the head and neck region of a newborn piglet (crossbred Large White and Landrace). The piglet was delivered by an apparently healthy sow in a litter with another nine healthy piglets. There were no large papillary tumorous lesions in the skin of any other animals on the breeding farm; however, there were multiple small warts on the external genitalia of several boars.The piglet was born with a cauliflower-like skin tumor the size of a walnut (3 ϫ 2 cm) behind the left ear. The tumorous mass grew rapidly, and a malignant cutaneous neoplasm was suspected. The piglet was killed at 42 days of age with barbiturate (Thiopental, Spofa, Prague). Samples for histopathology and ultrastructural pathology were collected immediately after the animal was killed. The material was fixed in 10% buffered formalin. Fixed tissues were processed for light microscopy using standard methods. Paraffin sections were stained with hematoxylin and eosin. Immunohistochemical studies of proliferation in the tumor were performed on formalin-fixed, paraffin-embedded tissue sections using the monoclonal antibody MIB1 against the Ki-67 antigen (dilution 1 : 100; Immunotech, Marseille, France) as described previously. 5 For transmission electron microscopy, the tissue samples fixed in 10% buffered formalin were washed in phosphate buffer, postfixed in 1% OsO 4 , dehydrated in graded ethanols, and embedded in Durcupan resin. Ultrathin sections were poststained with uranyl acetate and lead citrate and examined at 80 kV with a Philips EM 420 electron microscope.Grossly, an irregular 10 ϫ 15 cm, rough, cauliflower-like mass extended above the skin surface near the left ear and adjacent head and neck region of the newborn piglet (Fig. 1). There was no evidence of metastasis or lesions in the internal organs at autopsy.

Nasopharyngeal carcinoma (NPC) is a rare malignancy in the Czech Republic and Slovakia, with the standardized incidence rate of < 1:100000 person-years. Viral status of NPC in these non-endemic Eastern European regions is currently unknown. In a retrospective study, we evaluated the presence of EBV and HPV in 62 NPC cases. EBV status was determined by the use of in situ hybridization (ISH) for EBV encoded small RNA 1 (EBER1). HPV status was examined with p16 immunohistochemistry, DNA ISH and DNA polymerase chain reaction. Sixty-one studied cases showed non-keratinizing morphology and one was keratinizing squamous cell carcinoma. Only one NPC with non-keratinizing morphology was scored as p16-positive (nuclear and cytoplasmic staining ≥ 70% of tumor cells). This case was positive for high-risk HPV by ISH and the DNA PCR confirmed the presence of HPV18 type. At the same time, this case was found negative for EBV. Remaining sixty-one cases that were scored as p16-negative were all found HPV-negative by ISH and the DNA PCR. EBV was detected in 85.5% (53/62) of cases and 9 cases were EBV-negative, including the case of keratinizing NPC. In contrast with previous reports on the prevalence of EBV-positivity in Caucasian patients with NPC, the majority of patients coming from this non-endemic region show EBV-positivity; therefore, they may be candidates for novel EBV-targeting therapies. Conversely, HPV-positive NPC is very rare and HPV does not seem to play a significant role in the etiopathogenesis of NPC in these Eastern European populations.