Trimethoprim-sulfamethoxazole (SXT) is the preferable treatment option of the infections caused by Achromobacter spp. Our study aimed to analyze the SXT resistance of 98 Achromobacter spp. isolates from pediatric patients, among which 33 isolates were SXT-resistant. The presence of intI1 was screened by PCR and genome sequence analyses. The intI1 gene was detected in 10 of SXT-resistant isolates that had shorter intI1 PCR fragments named intI1S. Structural changes in intI1S were confirmed by genome sequencing and analyses which revealed 86 amino acids deletion in IntI1S protein compared to canonical IntI1 protein. All IntI1S isolates were of non-CF origin. Pan-genome analysis of intI1S bearing A. xylosoxidans isolates comprised 9052 genes, with the core genome consisting of 5455 protein-coding genes. Results in this study indicate that IntI1S isolates were derived from clinical settings and that cystic fibrosis (CF) patients were potential reservoirs for healthcare-associated infections that occurred in non-CF patients.

• Keywords
• Achromobacter spp., Class 1 integron integrase, Genome comparison,
• MeSH
• Achromobacter denitrificans * genetics   MeSH
• Achromobacter * MeSH
• Anti-Bacterial Agents therapeutic use   MeSH
• Cystic Fibrosis * MeSH
• Child MeSH
• Genomics MeSH
• Gram-Negative Bacterial Infections * MeSH
• Integrases therapeutic use   MeSH
• Integrons genetics   MeSH
• Trimethoprim, Sulfamethoxazole Drug Combination MeSH
• Humans MeSH
• Microbial Sensitivity Tests MeSH
• Check Tag
• Child MeSH
• Humans MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Serbia MeSH
• Names of Substances
• Anti-Bacterial Agents MeSH
• Integrases MeSH
• Trimethoprim, Sulfamethoxazole Drug Combination MeSH

In 2003 to 2004, the first five VIM-2 metallo-β-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.

• Keywords
• IncP-2, MBL, Pseudomonas aeruginosa, VIM, carbapenemase, megaplasmid, metallo-β-lactamase, plasmid,
• MeSH
• Anti-Bacterial Agents pharmacology   MeSH
• Bacterial Proteins MeSH
• beta-Lactamases genetics   metabolism   MeSH
• Epidemics * MeSH
• Phylogeny MeSH
• Cross Infection * epidemiology   MeSH
• Integrons genetics   MeSH
• Humans MeSH
• Multilocus Sequence Typing MeSH
• Hospitals MeSH
• Pseudomonas Infections * drug therapy   epidemiology   MeSH
• Pseudomonas aeruginosa genetics   metabolism   MeSH
• Electrophoresis, Gel, Pulsed-Field MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Poland MeSH
• Names of Substances
• Anti-Bacterial Agents MeSH
• Bacterial Proteins MeSH
• beta-Lactamases MeSH
• carbapenemase MeSH Browser

A VIM-1-producing ST92 Enterobacter cloacae was isolated in a Czech hospital. blaVIM-1 was part of the class 1 integron In110 carried by a Tn1721-like transposon. Tn1721-like was located on a ColE1-like plasmid, pEncl-30969cz (33,003 bp). Target site duplications at the boundaries of Tn1721-like suggested its transposition into the pEncl-30969cz backbone.

• Keywords
• Class 1 integron, Enterobacteriaceae, In110, Metallo-β-lactamase,
• MeSH
• Anti-Bacterial Agents pharmacology   MeSH
• Bacterial Proteins genetics   MeSH
• beta-Lactamases genetics   MeSH
• beta-Lactam Resistance genetics   MeSH
• DNA, Bacterial analysis   genetics   MeSH
• Enterobacter cloacae genetics   MeSH
• Enterobacteriaceae Infections microbiology   MeSH
• Integrons genetics   MeSH
• Humans MeSH
• Microbial Sensitivity Tests MeSH
• Plasmids genetics   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Anti-Bacterial Agents MeSH
• Bacterial Proteins MeSH
• beta-Lactamases MeSH
• DNA, Bacterial MeSH
• ROM protein, Bacteria MeSH Browser
• VIM-1 metallo-beta-lactamase MeSH Browser

ST252 Enterobacter cloacae, producing GES-5 carbapenemase, was isolated in a Czech hospital. blaGES-5 was part of a novel class 1 integron, In1406, which also included a new allele of the aadA15 gene cassette. In1406 was located on a ColE2-like plasmid, pEcl-35771cz (6953bp).

• Keywords
• Class 1 integron, Class A β-lactamase, ColE2-like, Enterobacteriaceae, In1406,
• MeSH
• Bacterial Proteins genetics   MeSH
• beta-Lactamases genetics   MeSH
• Enterobacter cloacae enzymology   genetics   MeSH
• Enterobacteriaceae Infections microbiology   MeSH
• Integrons genetics   MeSH
• Humans MeSH
• Multilocus Sequence Typing MeSH
• Hospitals MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Czech Republic MeSH
• Names of Substances
• Bacterial Proteins MeSH
• beta-Lactamases MeSH
• carbapenemase MeSH Browser

The objective of this study was to perform molecular surveillance for assessing the spread of carbapenemase-producing Pseudomonas aeruginosa in Czech hospitals. One hundred thirty-six carbapenemase-producing isolates were recovered from 22 hospitals located throughout the country. Sequence type 357 (ST357) dominated (n = 120) among carbapenemase producers. One hundred seventeen isolates produced IMP-type (IMP-7 [n = 116] and IMP-1 [n = 1]) metallo-β-lactamases (MβLs), 15 produced the VIM-2 MβL, and the remaining isolates expressed the GES-5 enzyme. The blaIMP-like genes were located in three main integron types, with In-p110-like being the most prevalent (n = 115). The two other IMP-encoding integrons (In1392 and In1393) have not been described previously. blaVIM-2-carrying integrons included In59-like, In56, and a novel element (In1391). blaGES-5 was carried by In717. Sequencing data showed that In-p110-like was associated with a Tn4380-like transposon inserted in genomic island LESGI-3 in the P. aeruginosa chromosome. The other integrons were also integrated into the P. aeruginosa chromosome. These findings indicated the clonal spread of ST357 P. aeruginosa, carrying the IMP-7-encoding integron In-p110, in Czech hospitals. Additionally, the sporadic emergence of P. aeruginosa producing different carbapenemase types, associated with divergent or novel integrons, punctuated the ongoing evolution of these bacteria.

• Keywords
• GES, Illumina sequencing, ST111, ST235, VIM, class 1 integrons, genomic islands (GIs), integrative conjugative element (ICE),
• MeSH
• Anti-Bacterial Agents pharmacology   MeSH
• Chromosomes, Bacterial chemistry   MeSH
• beta-Lactamases genetics   metabolism   MeSH
• Epidemiological Monitoring MeSH
• Gene Expression MeSH
• Genomic Islands MeSH
• Genotype MeSH
• Incidence MeSH
• Integrons MeSH
• Isoenzymes genetics   metabolism   MeSH
• Carbapenems pharmacology   MeSH
• Humans MeSH
• Microbial Sensitivity Tests MeSH
• Drug Resistance, Multiple, Bacterial genetics   MeSH
• Multilocus Sequence Typing MeSH
• Hospitals MeSH
• Pseudomonas Infections epidemiology   microbiology   MeSH
• Pseudomonas aeruginosa drug effects   enzymology   genetics   isolation & purification   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Multicenter Study MeSH
• Geographicals
• Czech Republic epidemiology   MeSH
• Names of Substances
• Anti-Bacterial Agents MeSH
• beta-lactamase IMP-7 protein, Pseudomonas aeruginosa MeSH Browser
• beta-Lactamases MeSH
• Isoenzymes MeSH
• Carbapenems MeSH

The aim of this study was to analyze the occurrence of sulfonamide resistance genes (sul1-3) and other genetic elements as antiseptic resistance gene (qacEΔ1) and class 1 and class 2 integrons (int1-2) in the upper layer of substrate and in the effluent of microcosm constructed wetlands (CWs) treating artificial wastewater containing diclofenac and sulfamethoxazole (SMX), which is a sulfonamide antibiotic. The bacteria in the substrate and in the effluents were equipped with the sul1-2, int1, and qacEΔ1 resistance determinants, which were introduced into the CW system during inoculation with activated sludge and with the soil attached to the rhizosphere of potted seedlings of Phalaris arundinacea 'Picta' roots (int1). By comparing the occurrence of the resistance determinants in the upper substrate layer and the effluent, it can be stated that they neither were lost nor emerged along the flow path. The implications of the presence of antibiotic resistance genes in the effluent may entail a risk of antibiotic resistance being spread in the receiving environment. Additionally, transformation products of SMX were determined. According to the obtained results, four (potential) SMX transformation products were identified. Two major metabolites of SMX, 2,3,5-trihydroxy-SMX and 3,5-dihydroxy-SMX, indicated that SMX may be partly oxidized during the treatment. The remaining two SMX transformation products (hydroxy-glutathionyl-SMX and glutathionyl-SMX) are conjugates with glutathione, which suggests the ability of CW bacterial community to degrade SMX and resist antimicrobial stress.

• Keywords
• Antibiotic resistance genes, Pharmaceuticals, Transformation products, Treatment wetlands,
• MeSH
• Anti-Bacterial Agents analysis   MeSH
• Drug Resistance, Microbial * MeSH
• Biodegradation, Environmental MeSH
• Integrons MeSH
• Pharmaceutical Preparations MeSH
• Wetlands MeSH
• Wastewater * MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Anti-Bacterial Agents MeSH
• Pharmaceutical Preparations MeSH
• Waste Water * MeSH

Two multidrug resistance (MDR) plasmids, carrying the VIM-1-encoding integron In110, were characterized. Plasmid pLec-476cz (311,758 bp), from a Leclercia adecarboxylata isolate, consisted of an IncHI1 backbone, a MDR region, and two accessory elements. Plasmid pKpn-431cz (142,876 bp), from a sequence type 323 (ST323) Klebsiella pneumoniae isolate, comprised IncFIIY-derived and pKPN3-like sequences and a mosaic region. A 40,400-bp sequence of pKpn-431cz was identical to the MDR region of pLec-476cz, indicating the en bloc acquisition of the VIM-1-encoding region from one plasmid by the other.

• Keywords
• IncFIIY, IncHI1, carbapenemases, integrative conjugative elements, metallo-β-lactamases,
• MeSH
• Bacterial Proteins genetics   MeSH
• beta-Lactamases genetics   MeSH
• beta-Lactam Resistance genetics   MeSH
• DNA, Bacterial genetics   MeSH
• Integrons genetics   MeSH
• Klebsiella pneumoniae drug effects   genetics   MeSH
• Humans MeSH
• Drug Resistance, Multiple, Bacterial genetics   MeSH
• Plasmids genetics   MeSH
• Base Sequence MeSH
• Sequence Analysis, DNA MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Czech Republic MeSH
• Names of Substances
• Bacterial Proteins MeSH
• beta-Lactamases MeSH
• carbapenemase MeSH Browser
• DNA, Bacterial MeSH
• VIM-1 metallo-beta-lactamase MeSH Browser

The complete nucleotide sequences of three multidrug resistance (MDR) IncA/C-like plasmids from Enterobacteriaceae isolates carrying the VIM-type carbapenemase-encoding integrons In4863 (blaVIM-19-aacA7-dfrA1-ΔaadA1-smr2) or In4873 (blaVIM-1-aacA7-dfrA1-ΔaadA1-smr2) were determined, which are the first In416-like elements identified in Greece. Plasmids pKP-Gr642 and pKP-Gr8143 were from Klebsiella pneumoniae ST383 isolates, whereas plasmid pEcl-Gr4873 was from an Enterobacter cloacae ST88 isolate. Sequencing showed that pKP-Gr642 (162787bp) and pKP-Gr8143 (154395bp) consisted of the type 1 IncA/C2 conserved backbone, the blaCMY-2-like gene-containing region, and the ARI-B (with the sul2 gene) and ARI-A (with a class 1 integron) resistance islands, like the plasmid pUMNK88_161 from the USA. The third plasmid, pEcl-Gr4873 (153958bp), exhibited extensive similarity with the type 2 IncA/C2 plasmid pR55 from France. pEcl-Gr4873 carried only one resistance island of a hybrid transposon structure inserted in a different location to ARI-A in type 1 A/C2 plasmids. In all three plasmids, the In416-like integrons In4863 or In4873 were identified within non-identical class II transposon structures. All three In416-like-carrying regions presented significant similarities with the MDR region of the IncA/C2 plasmid pCC416 from Italy, carrying the prototype In416 integron (blaVIM-4-aacA7-dfrA1-ΔaadA1-smr2). These findings provided the basis for speculations regarding the evolution of IncA/C2 plasmids with In416-like integrons, and confirmed the rapid evolution of some IncA/C2 plasmid lineages. Considering the broad host range of IncA/C2 molecules, it seems that pKP-Gr642, pKP-Gr8143 and pEcl-Gr4873 plasmids might support the diffusion of In416-like integrons among Enterobacteriaceae.

• Keywords
• CMY, Class 1 integron, IncA/C(2) plasmids, VIM,
• MeSH
• beta-Lactamases genetics   MeSH
• Enterobacter cloacae enzymology   genetics   isolation & purification   MeSH
• Enterobacteriaceae Infections microbiology   MeSH
• Integrons * MeSH
• Klebsiella pneumoniae enzymology   genetics   isolation & purification   MeSH
• Humans MeSH
• Molecular Sequence Data MeSH
• Plasmids * MeSH
• Gene Order MeSH
• Sequence Analysis, DNA MeSH
• Sequence Homology MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Greece MeSH
• Names of Substances
• beta-Lactamases MeSH

International trade with ornamental fish is gradually recognized as an important source of a wide range of different antibiotic resistant bacteria. In this study we therefore characterized the prevalence of selected antibiotic resistance genes in the microbiota found in the carriage water of ornamental fish originating from 3 different continents. Real-time PCR quantification showed that the sul1 gene was present in 11 out of 100 bacteria. tet(A) was present in 6 out of 100 bacteria and strA, tet(G), sul2 and aadA were present in 1-2 copies per 100 bacteria. Class I integrons were quite common in carriage water microbiota, however, pyrosequencing showed that only 12 different antibiotic gene cassettes were present in class I integrons. The microbiota characterized by pyrosequencing of the V3/V4 variable region of 16S rRNA genes consisted of Proteobacteria (48%), Bacteroidetes (29.5%), Firmicutes (17.8%), Actinobacteria (2.1%) and Fusobacteria (1.6%). Correlation analysis between antibiotic resistance gene prevalence and microbiota composition verified by bacterial culture showed that major reservoirs of sul1 sul2, tet(A), tet(B) tet(G), cat, cml, bla, strA, aacA, aph and aadA could be found among Alpha-, Beta- and Gammaproteobacteria with representatives of Enterobacteriaceae, Pseudomonadaceae, Rhizobiaceae and Comamonadaceae being those most positively associated with the tested antibiotic resistance genes.

• MeSH
• Actinobacteria drug effects   genetics   MeSH
• Anti-Bacterial Agents pharmacology   MeSH
• Bacteria drug effects   genetics   MeSH
• Bacteroidetes drug effects   genetics   MeSH
• Fusobacteria drug effects   genetics   MeSH
• Integrons genetics   MeSH
• Real-Time Polymerase Chain Reaction MeSH
• Water Microbiology MeSH
• Microbiota drug effects   genetics   MeSH
• Drug Resistance, Multiple, Bacterial MeSH
• Proteobacteria drug effects   genetics   MeSH
• RNA, Ribosomal, 16S genetics   MeSH
• Fishes MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Anti-Bacterial Agents MeSH
• RNA, Ribosomal, 16S MeSH

Carbapenemase-producing Enterobacteriaceae and Pseudomonas spp. are increasingly reported in many countries all over the world. Due to the resistance of those bacteria to almost all antibiotics (e.g.beta-lactams, aminoglycosides, fluoroquinolones),treatment options are seriously limited. In the Czech Republic, the incidence of carbapenemase-producing Enterobacteriaceae seems to be low, restricted to only three cases detected between 2009 and 2010.Here, we describe molecular typing of 15 carbapenemase-producing Klebsiella pneumoniae isolates identified in the Czech Republic during 2011. Five VIM-1-producing isolates belonging to sequence type (ST)11 and one VIM-4-producing isolate of ST1029 have been detected. blaVIM-1 and blaVIM-4 as a part of class 1 integrons were chromosomally located or carried by a plasmid belonging to A/C replicon type (blaVIM-4). KPC-3-producing isolates of ST512, recovered from six patients, caused an outbreak. Three more isolates producing KPC-2 enzyme belonged to ST258. Both blaKPCgenes were part of the Tn4401a transposon carried on plasmids of the pKpQIL type. The isolates were resistant to all antibiotics tested except colistin and/or gentamicin.Four of these 15 strains were recovered from patients repatriated to the Czech Republic from Greece and Italy. This is the first report of outbreaks caused by carbapenemase-producing Enterobacteriaceae in the Czech Republic.

• MeSH
• Anti-Bacterial Agents pharmacology   MeSH
• Bacterial Proteins metabolism   MeSH
• beta-Lactamases metabolism   MeSH
• beta-Lactam Resistance MeSH
• Travel MeSH
• Incidence MeSH
• Klebsiella Infections diagnosis   enzymology   epidemiology   MeSH
• Integrons genetics   MeSH
• Carbapenems pharmacology   MeSH
• Klebsiella pneumoniae classification   drug effects   genetics   isolation & purification   MeSH
• Humans MeSH
• Microbial Sensitivity Tests MeSH
• Drug Resistance, Multiple, Bacterial MeSH
• Molecular Typing MeSH
• Plasmids genetics   metabolism   MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Czech Republic epidemiology   MeSH
• Italy MeSH
• Greece MeSH
• Names of Substances
• Anti-Bacterial Agents MeSH
• Bacterial Proteins MeSH
• beta-Lactamases MeSH
• carbapenemase MeSH Browser
• Carbapenems MeSH