scholarly journals Undetectable Production of the VIM-1 Carbapenemase in an Atlantibacter hermannii Clinical Isolate

2021 ◽  
Vol 12 ◽  
Author(s):  
Delphine Girlich ◽  
Rémy A. Bonnin ◽  
Alexis Proust ◽  
Thierry Naas ◽  
Laurent Dortet
Keyword(s):  
Rectal Swab ◽  
Selective Pressure ◽  
Resistance Pattern ◽  
Read Coverage ◽  
Rt Pcr ◽  
Incompatibility Group ◽  
Class 1 Integron ◽  
Class 1 ◽  
Gene Transcripts ◽  
Enterobacter Hormaechei

The differential expression of VIM-1 in Atlantibacter hermannii WEB-2 and Enterobacter hormaechei ssp. hoffmannii WEB-1 clinical isolates from a rectal swab of a hospitalized patient in France was investigated. A. hermannii WEB-2 was resistant to all β-lactams except carbapenems. It produced ESBL SHV-12, but the Carba NP test failed to detect any carbapenemase activity despite the production of VIM-1. Conversely, E. hormaechei WEB-1, previously recovered from the same patient, was positive for the detection of carbapenemase activity. The blaVIM–1 gene was located on a plasmid and embedded within class 1 integron. Both plasmids were of the same IncA incompatibility group and conferred the same resistance pattern when electroporated in Escherichia coli TOP10 or Enterobacter cloacae CIP7933. Quantitative RT-PCR experiments indicated a weaker replication of pWEB-2 in A. hermannii as compared to E. hormaechei. An isogenic mutant of A. hermannii WEB-2 selected after sequential passages with increased concentrations of imipenem possessed higher MICs for carbapenems and cephalosporins including cefiderocol, higher levels of the blaVIM–1 gene transcripts, and detectable carbapenemase activity using the Carba NP test. Assessment of read coverage demonstrated that a duplication of the region surrounding blaVIM–1 gene occurred in the A. hermannii mutant with detectable carbapenemase activity. The lack of detection of the VIM-1 carbapenemase activity in A. hermannii WEB-2 isolate was likely due to a weak replication of the IncA plasmid harboring the blaVIM–1 gene. Imipenem as selective pressure led to a duplication of this gene on the plasmid and to the restoration of a significant carbapenem-hydrolyzing phenotype.

scholarly journals A Predominant Multidrug-Resistant Salmonella enterica Serovar Saintpaul Clonal Line in German Turkey and Related Food Products

2010 ◽  
Vol 76 (11) ◽  
pp. 3657-3667 ◽  
Author(s):  
Janine Beutlich ◽  
Irene Rodr�guez ◽  
Andreas Schroeter ◽  
Annemarie K�sbohrer ◽  
Reiner Helmuth ◽  
...  
Keyword(s):  
Salmonella Enterica ◽  
Food Products ◽  
Gene Cassette ◽  
Multidrug Resistant ◽  
Resistance Pattern ◽  
Clonal Line ◽  
Class 1 Integron ◽  
Pathogenic Potential ◽  
Class 1 ◽  
Plasmid Profiling

ABSTRACT Recently, Salmonella enterica subsp. enterica serovar Saintpaul has increasingly been observed in several countries, including Germany. However, the pathogenic potential and epidemiology of this serovar are not very well known. This study describes biological attributes of S. Saintpaul isolates obtained from turkeys in Germany based on characterization of their pheno- and genotypic properties. Fifty-five S. Saintpaul isolates from German turkeys and turkey-derived food products isolated from 2000 to 2007 were analyzed by using antimicrobial agent, organic solvent, and disinfectant susceptibility tests, isoelectric focusing, detection of resistance determinants, plasmid profiling, pulsed-field gel electrophoresis (PFGE), and hybridization experiments. These isolates were compared to an outgroup consisting of 24 S. Saintpaul isolates obtained from humans and chickens in Germany and from poultry and poultry products (including turkeys) in Netherlands. A common core resistance pattern was detected for 27 German turkey and turkey product isolates. This pattern included resistance (full or intermediate) to ampicillin, amoxicillin-clavulanic acid, gentamicin, kanamycin, nalidixic acid, streptomycin, spectinomycin, and sulfamethoxazole and intermediate resistance or decreased susceptibility to ciprofloxacin (MIC, 2 or 1 μg/ml, respectively) and several third-generation cephalosporins (including ceftiofur and cefoxitin [MIC, 4 to 2 and 16 to 2 μg/ml, respectively]). These isolates had the same core resistance genotype, with bla TEM-1, aadB, aadA2, sul1, a Ser83→Glu83 mutation in the gyrA gene, and a chromosomal class 1 integron carrying the aadB-aadA2 gene cassette. Their XbaI, BlnI, and combined XbaI-BlnI PFGE patterns revealed levels of genetic similarity of 93, 75, and 90%, respectively. This study revealed that a multiresistant S. Saintpaul clonal line is widespread in turkeys and turkey products in Germany and was also detected among German human fecal and Dutch poultry isolates.

scholarly journals Genome and Plasmid Analysis ofblaIMP-4-Carrying Citrobacter freundii B38

2016 ◽  
Vol 60 (11) ◽  
pp. 6719-6725 ◽  
Author(s):  
Jianhui Xiong ◽  
Maxime Déraspe ◽  
Naeem Iqbal ◽  
Jennifer Ma ◽  
Frances B. Jamieson ◽  
...  
Keyword(s):  
Gram Negative Bacteria ◽  
Incompatibility Group ◽  
Class 1 Integron ◽  
Evolutionary Pathway ◽  
Content Type ◽  
Conserved Sequence ◽  
Distinct Evolutionary Lineage ◽  
Plasmid Analysis ◽  
Class 1 ◽  
The Western Pacific

ABSTRACTSequencing of theblaIMP-4-carryingC. freundiiB38 using the PacBio SMRT technique revealed that the genome contained a chromosome of 5,134,500 bp and three plasmids, pOZ172 (127,005 bp), pOZ181 (277,592 bp), and pOZ182 (18,467 bp). Plasmid pOZ172 was identified as IncFIIY, like pP10164-NDM and pNDM-EcGN174. It carries a class 1 integron with four cassettes (blaIMP-4-qacG2-aacA4-aphA15) and a complete hybridtnimodule (tniR-tniQ-tniB-tniA). The recombination oftniRfrom Tn402(identical) withtniQBAfrom Tn5053(99%) occurred within theressite of Tn402/5053. The Tn402/5053-like integron, named Tn6017, was inserted into Tn1722at theresII site. The replication, partitioning, and transfer systems of pOZ181 were similar to those of IncHI2 plasmids (e.g., R478) and contained asul1-type class 1 integron with the cassette arrayorf-dfrA1-orf-gcu37-aadA5linked to an upstream Tn1696 tnpA-tnpRand to a downstream 3′ conserved sequence (3′-CS) and ISCR1. A Tn2transposon encoding ablaTEM-1β-lactamase was identified on pOZ182. Other interesting resistance determinants encoded on the B38 chromosome included multidrug resistance (MDR) efflux pumps, an AmpC β-lactamase, and resistances to Cu, Ag, As, and Zn. This is the first report of a completetnimodule linked to ablaIMP-4-carrying class 1 integron, which, together with other recently reported non-sul1integrons, represents the emergence of a distinct evolutionary lineage of class 1 integrons lacking a 3′-CS (qacEΔ1-sul1). The unique cassette array, completetnimodule of Tn6017, and incompatibility group of pOZ172 suggest ablaIMP-4evolutionary pathway inC. freundiiB38 different from that for otherblaIMP-4genes found in Gram-negative bacteria in the Western Pacific region.

scholarly journals Evaluation of Class 1 and 2 Integrons and Antibiotic Resistance Pattern in Salmonella enterica Isolated from Diarrheal Food-Borne Outbreaks in Iran

2019 ◽  
Author(s):  
S.F. Sayadnouri ◽  
M.M. Soltan Dallal ◽  
S. Akbarzadeh ◽  
R. Mazaheri Nezhad Fard
Keyword(s):  
Antibiotic Resistance ◽  
Salmonella Enterica ◽  
Control Measures ◽  
Resistance Pattern ◽  
Salmonella Spp ◽  
Chi Square ◽  
Class 1 Integron ◽  
Antibiotic Resistance Pattern ◽  
Food Borne ◽  
Class 1

Background: Salmonella spp. are major causes of food-borne disease and have been identified among many diarrheal outbreaks. The major aim of the current investigation was to evaluate the class 1 and 2 integrons and antibiotic resistance pattern in Salmonella enterica isolated from diarrheal food-borne outbreaks in Iran.  Methods: This study was carried out on 115 diarrheal feces samples obtained from food-borne outbreak in 2016 in Iran. Antimicrobial resistance patterns of 27 isolated S. enterica seovars and presence of class 1 and class 2 integrons in the serovars were investigated using conventional and molecular methods. Results were statistically analyzed using SPSS software v. 21 and Chi-Square test. Results: Overall, 27 S. enterica were characterized as 14 S. Paratyphi C, 7 S. Enteritidis, 5 S. Paratyphi D, and 1 S. Paratyphi A serovars. Results of molecular assay showed that class 1 integron presented in all and class 2 integron in three strains. All isolates with class 2 integron genes were resistant to almost all the antimicrobials. Conclusion: Most studied Salmonella strains from diarrheal food-borne outbreak of Iran in 2016 were multiple resistant to the highlighted antimicrobials. Knowledge about risk factor involving the salmonellosis and their control measures could help the national authorities to prevent the outbreaks. Further comprehensive studies with larger sample sizes are necessary to acquire more data about risk factors of multiple resistant Salmonella outbreaks in the country.

scholarly journals A Novel, Integron-Regulated, Class C β-Lactamase

Antibiotics ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 123
Author(s):  
Maria-Elisabeth Böhm ◽  
Mohammad Razavi ◽  
Carl-Fredrik Flach ◽  
D. G. Joakim Larsson
Keyword(s):  
Bacterial Infections ◽  
Gene Cassette ◽  
Fourth Generation ◽  
Resistance Pattern ◽  
Asian Countries ◽  
Class 1 Integron ◽  
Class 1 ◽  
Resistant Strains ◽  
Class C ◽  
Mobile Context

AmpC-type β-lactamases severely impair treatment of many bacterial infections, due to their broad spectrum (they hydrolyze virtually all β-lactams, except fourth-generation cephalosporins and carbapenems) and the increasing incidence of plasmid-mediated versions. The original chromosomal AmpCs are often tightly regulated, and their expression is induced in response to exposure to β-lactams. Regulation of mobile ampC expression is in many cases less controlled, giving rise to constitutively resistant strains with increased potential for development or acquisition of additional resistances. We present here the identification of two integron-encoded ampC genes, blaIDC-1 and blaIDC-2 (integron-derived cephalosporinase), with less than 85% amino acid sequence identity to any previously annotated AmpC. While their resistance pattern identifies them as class C β-lactamases, their low isoelectric point (pI) values make differentiation from other β-lactamases by isoelectric focusing impossible. To the best of our knowledge, this is the first evidence of an ampC gene cassette within a class 1 integron, providing a mobile context with profound potential for transfer and spread into clinics. It also allows bacteria to adapt expression levels, and thus reduce fitness costs, e.g., by cassette-reshuffling. Analyses of public metagenomes, including sewage metagenomes, show that the discovered ampCs are primarily found in Asian countries.

scholarly journals Molecular Characterization of a Novel Class 1 Integron Containing blaGES-1 and a Fused Product of aac(3)-Ib/aac(6")-Ib" Gene Cassettes in Pseudomonas aeruginosa

2002 ◽  
Vol 46 (3) ◽  
pp. 638-645 ◽  
Author(s):  
Véronique Dubois ◽  
Laurent Poirel ◽  
Caroline Marie ◽  
Corinne Arpin ◽  
Patrice Nordmann ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Diffusion Method ◽  
Fusion Product ◽  
Resistance Pattern ◽  
Clinical Strain ◽  
Homologous Sequence ◽  
Class 1 Integron ◽  
Gene Cassettes ◽  
Extended Spectrum ◽  
Class 1

ABSTRACT As seen by the disk diffusion method, the clinical strain of Pseudomonas aeruginosa Pa695, resistant to all extended-spectrum cephalosporins and aminoglycosides, exhibited an unusual synergistic effect between ceftazidime and imipenem. This isolate produced an extended-spectrum β-lactamase (ESBL) with a pI of 5.8 that appeared to be chromosomally encoded. Cloning experiments revealed that this ESBL was encoded by bla GES-1, previously described in an integron from Klebsiella pneumoniae. In P. aeruginosa Pa695, a higher level of resistance to ceftazidime than to ticarcillin was observed, and no synergy between the β-lactamase inhibitors and extended-spectrum cephalosporins was detected, in contrast to the resistance pattern observed in K. pneumoniae. Further sequence analysis demonstrated that the bla GES-1 gene cassette was located in a class 1 integron, which contained another sequence corresponding to the fused aac(3)-Ib and aac(6")-Ib" gene cassettes. The fusion product was functional, as was the product of each gene cloned separately: AAC(3)-I, despite the deletion of the four last amino acids, and AAC(6"), which carried three amino acid changes compared with the most homologous sequence. The AAC(3)-I protein conferred an expected gentamicin and fortimicin resistance, and the AAC(6"), despite the Leu-119→Ser substitution, yielded resistance to kanamycin, tobramycin, and dibekacin, but slightly affected netilmicin and amikacin, and had no apparent effect on gentamicin. The fusion product conveyed a large profile of resistance, combining the AAC(6") activity with a higher level of gentamicin resistance without accompanying fortimicin resistance.

scholarly journals Exploring the global spread of Klebsiella grimontii isolates possessing bla VIM-1 and mcr-9

2021 ◽  
Author(s):  
Edgar I. Campos-Madueno ◽  
Aline I. Moser ◽  
Martin Risch ◽  
Bodmer Thomas ◽  
Endimiani Andrea
Keyword(s):  
Core Genome ◽  
Klebsiella Oxytoca ◽  
Regulatory Elements ◽  
Class 1 Integron ◽  
Closely Related Species ◽  
Global Spread ◽  
Class 1 ◽  
Resistance Patterns ◽  
Enterobacter Hormaechei ◽  
Antibiotic Resistance Patterns

The spread of plasmid-mediated carbapenemases within Klebsiella oxytoca is well-documented. In contrast, data concerning the closely-related species Klebsiella grimontii are scarce. In fact, despite the recent report of the first bla KPC-2 -producing K. grimontii , nothing is known about its clonality and antibiotic resistance patterns. In a retrospective search in our collection, we identified 2 bla VIM -positive K. oxytoca strains. Whole-genome sequencing with both Illumina and Nanopore indicated that our strains actually belonged to K. grimontii and were of sequence type (ST) 172 and ST189. Moreover, the two strains were associated with 297kb IncHI2/HI2A-pST1 and 90.6kb IncFII(Yp) plasmids carrying bla VIM-1 together with mcr-9 and bla VIM-1 , respectively. In the IncHI2/HI2A plasmid, bla VIM-1 was located in a class 1 integron (In 110 ), while mcr-9 was associated to the qseC - qseB- like regulatory elements. Overall, this plasmid showed to be very similar to those carried by other Enterobacterales isolated from food and animal sources (e.g., Salmonella and Enterobacter spp. detected in Germany and Egypt). The IncFII(Yp) plasmid was unique and its bla VIM-1 region was associated to a rare integron (In 1373 ). Mapping of In 1373 indicated a possible origin in Austria from an Enterobacter hormaechei carrying a highly similar plasmid. Core-genome phylogenies indicated that the ST172 K. grimontii belonged to a clone of identical Swedish and Swiss strains (≤15 SNVs to each other), whereas the ST189 strain was sporadic. Surveillance of carbapenemase-producing K. oxytoca strains should be reinforced to detect and prevent the dissemination of new species belonging to the Klebsiella genus.

scholarly journals SMB-1, a Novel Subclass B3 Metallo-β-Lactamase, Associated with ISCR1and a Class 1 Integron, from a Carbapenem-Resistant Serratia marcescens Clinical Isolate

2011 ◽  
Vol 55 (11) ◽  
pp. 5143-5149 ◽  
Author(s):  
Jun-ichi Wachino ◽  
Hiroyuki Yoshida ◽  
Kunikazu Yamane ◽  
Satowa Suzuki ◽  
Mari Matsui ◽  
...  
Keyword(s):  
Serratia Marcescens ◽  
Clinical Isolate ◽  
Apple Orchard ◽  
Resistance Pattern ◽  
Binding Motif ◽  
Class 1 Integron ◽  
Uncultured Bacterium ◽  
Content Type ◽  
Carbapenem Resistant ◽  
Class 1

ABSTRACTA carbapenem-resistantSerratia marcescensstrain, 10mdr148, was identified in a Japanese hospital in 2010. The carbapenem resistance of this strain was attributed to the production of a novel metallo-β-lactamase (MBL), named SMB-1 (Serratiametallo-β-lactamase). SMB-1 possessed a zinc binding motif, H(Q)XHXDH (residues 116 to 121), H196, and H263 and was categorized as a member of subclass B3 MBL. SMB-1 has 75% amino acid identity with the most closely related MBL, AMO1, of uncultured bacterium, recently identified through the metagenomic analysis of apple orchard soil. The introduction ofblaSMB-1intoEscherichia coliconferred resistance to a variety of β-lactam antibiotics, penicillins, cephalosporins, and carbapenems, but not aztreonam, a resistance pattern consistent with those of other MBLs. SMB-1 demonstrated highkcatvalues of >500 s−1for carbapenems, resulting in the highest hydrolyzing efficiency (kcat/Km) among the agents tested. The hydrolyzing activity of SMB-1 was well inhibited by chelating agents. TheblaSMB-1gene was located on the chromosome ofS. marcescensstrain 10mdr148 and at the 3′ end of the ISCR1element in complex with a typical class 1 integron carryingaac(6′)-IbandcatB3gene cassettes. Downstream ofblaSMB-1, the second copy of the 3′conserved segment and ISCR1were found. To our knowledge, this is the first subclass B3 MBL gene associated with an ISCR1element identified in anEnterobacteriaceaeclinical isolate. A variety of antibiotic resistance genes embedded with ISCR1have been widely spread amongEnterobacteriaceaeclinical isolates, thus the further dissemination ofblaSMB-1mediated by ISCR1transposition activity may become a future concern.

scholarly journals Beta‐lactamase‐producing Escherichia coli in Bangladesh: Their phenotypic and molecular characteristics

2019 ◽  
Vol 28 (1) ◽  
pp. 71-81
Author(s):  
Sunjukta Ahsan ◽  
Riajul Islam
Keyword(s):  
Health Concern ◽  
Resistance Pattern ◽  
Molecular Characteristics ◽  
New Delhi ◽  
Beta Lactamase ◽  
Beta Lactam ◽  
Class 1 Integron ◽  
E Coli ◽  
Beta Lactam Antibiotics ◽  
Class 1

The emergence and rapid dissemination of beta-lactamase-producing E. coli is now a worldwide problem. A total of 45 E. coli obtained from clinical specimens from a medical service centre in Dhaka were selected for this study. Test E. coli exhibited variable resistance to 3rd (71.7 - 97.8%, n = 48) and 4th (78%, n = 48) generation beta-lactam antibiotics, with 72% sensitivity to Carbapenem. Analysis of co-resistance indicated that 33.3% of E. coli (n = 48) were co-resistant to beta-lactams and ciprofloxacin. ESBL producers were predominant comprising of 84.7% E. coli. Among them, 22.7% contained blaTEM, 24.2% contained blaCTX-M, 4.3% contained blaSHV and 9.1% contained blaOXA-1 genes. A total of 25.75% isolates were metallo beta-lactamase producers. Of these, 1.5% of E. coli strains contained New Delhi metallo beta-lactamase gene and 6% contained AmpC gene. Multiple beta-lactamase genes were detected in some test isolates; 6.7% isolates contained 4, 20% contained 3 and 73.3% contained 2 beta lactamase genes. Fifty per cent of the E. coli contained plasmids of variable sizes. In addition, a total of 39% of the E. coli contained Class 1 integron. The increasing trend in beta-lactam resistance is of public health concern as it limits treatment regime and indicates to the need of continuous monitoring of resistance pattern. Dhaka Univ. J. Biol. Sci. 28(1): 71-81, 2019 (January)

Detection of Nucleic Acids in Cells or Tissue Sections Using In situ Polymerase Chain Reaction (PCR)

1996 ◽  
Vol 54 ◽  
pp. 16-17
Author(s):  
J. R. Hully ◽  
K. R. Luehrsen ◽  
K. Aoyagi ◽  
C. Shoemaker ◽  
R. Abramson
Keyword(s):  
Nucleic Acids ◽  
Viral Infections ◽  
Anatomical Location ◽  
Rt Pcr ◽  
Reverse Transcription Pcr ◽  
Cellular Source ◽  
Gene Transcripts ◽  
Initial Description ◽  
In Cells

The development of PCR technology has greatly accelerated medical research at the genetic and molecular levels. Until recently, the inherent sensitivity of this technique has been limited to isolated preparations of nucleic acids which lack or at best have limited morphological information. With the obvious exception of cell lines, traditional PCR or reverse transcription-PCR (RT-PCR) cannot identify the cellular source of the amplified product. In contrast, in situ hybridization (ISH) by definition, defines the anatomical location of a gene and/or it’s product. However, this technique lacks the sensitivity of PCR and cannot routinely detect less than 10 to 20 copies per cell. Consequently, the localization of rare transcripts, latent viral infections, foreign or altered genes cannot be identified by this technique. In situ PCR or in situ RT-PCR is a combination of the two techniques, exploiting the sensitivity of PCR and the anatomical definition provided by ISH. Since it’s initial description considerable advances have been made in the application of in situ PCR, improvements in protocols, and the development of hardware dedicated to in situ PCR using conventional microscope slides. Our understanding of the importance of viral latency or viral burden in regards to HIV, HPV, and KSHV infections has benefited from this technique, enabling detection of single viral copies in cells or tissue otherwise thought to be normal. Clearly, this technique will be useful tool in pathobiology especially carcinogenesis, gene therapy and manipulations, the study of rare gene transcripts, and forensics.

Antimicrobial Resistance and Presence of Class 1 Integrons Among Different Serotypes of Salmonella spp. Recovered From Children with Diarrhea in Tehran, Iran

2020 ◽  
Vol 20 (2) ◽  
pp. 160-166
Author(s):  
Seyedeh Hanieh Eshaghi Zadeh ◽  
Hossein Fahimi ◽  
Fatemeh Fardsanei ◽  
Mohammad Mehdi Soltan Dallal
Keyword(s):  
Antibiotic Resistance ◽  
Antimicrobial Resistance ◽  
Multidrug Resistant ◽  
Salmonella Spp ◽  
Class 1 Integron ◽  
Class 1 Integrons ◽  
Food Borne ◽  
Class 1 ◽  
Resistance Patterns ◽  
C 30

Background: Salmonellosis is a major food-borne disease worldwide. The increasing prevalence of antimicrobial resistance among food-borne pathogens such as Salmonella spp. is concerning. Objective: The main objective of this study is to identify class 1 integron genes and to determine antibiotic resistance patterns among Salmonella isolates from children with diarrhea. Methods: A total of 30 Salmonella isolates were recovered from children with diarrhea. The isolates were characterized for antimicrobial susceptibility and screened for the presence of class 1 integron genes (i.e. intI1, sulI1, and qacEΔ1). Results: The most prevalent serotype was Enteritidis 36.7%, followed by Paratyphi C (30%), and Typhimurium (16.7%). The highest rates of antibiotic resistance were obtained for nalidixic acid (53.3%), followed by streptomycin (40%), and tetracycline (36.7%). Regarding class 1 integrons, 36.7%, 26.7%, and 33.3% of the isolates carried intI1, SulI, and qacEΔ1, respectively, most of which (81.8%) were multidrug-resistant (MDR). Statistical analysis revealed that the presence of class 1 integron was significantly associated with resistance to streptomycin and tetracycline (p = 0.042). However, there was no association between class 1 integron and other antibiotics used in this study (p > 0.05). Conclusion: The high frequency of integron class 1 gene in MDR Salmonella strains indicates that these mobile genetic elements are versatile among different Salmonella serotypes, and associated with reduced susceptibility to many antimicrobials.

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