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Suzuki, Haruo, Hirokazu Yano, Celeste J. Brown, and Eva M. Top. "Predicting Plasmid Promiscuity Based on Genomic Signature." Journal of Bacteriology 192, no. 22 (2010): 6045–55. http://dx.doi.org/10.1128/jb.00277-10.
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ABSTRACT Despite the important contribution of self-transmissible plasmids to bacterial evolution, little is understood about the range of hosts in which these plasmids have evolved. Our goal was to infer this so-called evolutionary host range. The nucleotide composition, or genomic signature, of plasmids is often similar to that of the chromosome of their current host, suggesting that plasmids acquire their hosts’ signature over time. Therefore, we examined whether the evolutionary host range of plasmids could be inferred by comparing their trinucleotide composition to that of all completely sequenced bacterial chromosomes. The diversity of candidate hosts was determined using taxonomic classification and genetic distance. The method was first tested using plasmids from six incompatibility (Inc) groups whose host ranges are generally thought to be narrow (IncF, IncH, and IncI) or broad (IncN, IncP, and IncW) and then applied to other plasmid groups. The evolutionary host range was found to be broad for IncP plasmids, narrow for IncF and IncI plasmids, and intermediate for IncH and IncN plasmids, which corresponds with their known host range. The IncW plasmids as well as several plasmids from the IncA/C, IncP, IncQ, IncU, and PromA groups have signatures that were not similar to any of the chromosomal signatures, raising the hypothesis that these plasmids have not been ameliorated in any host due to their promiscuous nature. The inferred evolutionary host range of IncA/C, IncP-9, and IncL/M plasmids requires further investigation. In this era of high-throughput sequencing, this genomic signature method is a useful tool for predicting the host range of novel mobile elements.
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Getino, María, David J. Sanabria-Ríos, Raúl Fernández-López, et al. "Synthetic Fatty Acids Prevent Plasmid-Mediated Horizontal Gene Transfer." mBio 6, no. 5 (2015): e01032-15. https://doi.org/10.1128/mBio.01032-15.
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Bacterial conjugation constitutes a major horizontal gene transfer mechanism for the dissemination of antibiotic resistance genes among human pathogens. Antibiotic resistance spread could be halted or diminished by molecules that interfere with the conjugation process. In this work, synthetic 2-alkynoic fatty acids were identified as a novel class of conjugation inhibitors. Their chemical properties were investigated by using the prototype 2-hexadecynoic acid and its derivatives. Essential features of effective inhibitors were the carboxylic group, an optimal long aliphatic chain of 16 carbon atoms, and one unsaturation. Chemical modification of these groups led to inactive or less-active derivatives. Conjugation inhibitors were found to act on the donor cell, affecting a wide number of pathogenic bacterial hosts, including <em>Escherichia</em>, <em>Salmonella</em>, <em>Pseudomonas</em>, and<em>Acinetobacter</em> spp. Conjugation inhibitors were active in inhibiting transfer of IncF, IncW, and IncH plasmids, moderately active against IncI, IncL/M, and IncX plasmids, and inactive against IncP and IncN plasmids. Importantly, the use of 2-hexadecynoic acid avoided the spread of a derepressed IncF plasmid into a recipient population, demonstrating the feasibility of abolishing the dissemination of antimicrobial resistances by blocking bacterial conjugation.
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Schwanbeck, Julian, Wolfgang Bohne, Ufuk Hasdemir, et al. "Detection of a New Resistance-Mediating Plasmid Chimera in a blaOXA-48-Positive Klebsiella pneumoniae Strain at a German University Hospital." Microorganisms 9, no. 4 (2021): 720. http://dx.doi.org/10.3390/microorganisms9040720.
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Mobile genetic elements, such as plasmids, facilitate the spread of antibiotic resistance genes in Enterobacterales. In line with this, we investigated the plasmid-resistome of seven blaOXA-48 gene-carrying Klebsiella pneumoniae isolates, which were isolated between 2013 and 2014 at the University Medical Center in Göttingen, Germany. All isolates were subjected to complete genome sequencing including the reconstruction of entire plasmid sequences. In addition, phenotypic resistance testing was conducted. The seven isolates comprised both disease-associated isolates and colonizers isolated from five patients. They fell into two clusters of three sequence type (ST)101 and two ST11 isolates, respectively; and ST15 and ST23 singletons. The seven isolates harbored various plasmids of the incompatibility (Inc) groups IncF, IncL/M, IncN, IncR, and a novel plasmid chimera. All blaOXA-48 genes were encoded on the IncL/M plasmids. Of note, distinct phenotypical resistance patterns associated with different sets of resistance genes encoded by IncL/M and IncR plasmids were observed among isolates of the ST101 cluster in spite of high phylogenetic relatedness of the bacterial chromosomes, suggesting nosocomial transmission. This highlights the importance of plasmid uptake and plasmid recombination events for the fast generation of resistance variability after clonal transmission. In conclusion, this study contributes a piece in the puzzle of molecular epidemiology of resistance gene-carrying plasmids in K. pneumoniae in Germany.
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Bihannic, Morgan, Marisa Haenni, Eric Oswald, and Jean-Yves Madec. "Divergent Evolution of the repFII Replicon of IncF Plasmids Carrying Cytotoxic Necrotizing Factorcnf2, Cytolethal Distending ToxincdtIII, andf17AeFimbrial Variant Genes in Type 2 Necrotoxigenic Escherichia coli Isolates from Calves." Applied and Environmental Microbiology 82, no. 2 (2015): 510–17. http://dx.doi.org/10.1128/aem.02641-15.
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ABSTRACTAmong the pathovars ofEscherichia coliin cattle, necrotoxigenicE. coli(NTEC) is defined by the production of cytotoxic necrotizing factors (CNFs). In particular, type 2 NTEC (NTEC2) strains are frequent in diarrheic and septicemic calves and usually coproduce CNF type 2 (CNF2), cytolethal distending toxin type III (CDTIII), and fimbrial adhesins of the F17 family, whose genetic determinants have frequently been reported on the same Vir-like plasmid. In this study, we investigated the genetic environment of thecnf2,f17Ae, andcdtIIIgenes in a collection of fecalE. coliisolates recovered from 484 French and 58 Iranian calves. In particular, we highlighted the spread ofcnf2,f17Ae, andcdtIIIon similar 150-kb IncF plasmids harboring the newly assigned repFII replicon allele F74 in NTEC2 isolates. Interestingly, this 150-kb IncF plasmid differed from the 140-kb IncF plasmid harboring the newly assigned repFII replicon allele F75 and carryingcnf2alone. These results suggest two divergent lineages ofcnf2-carrying IncF plasmids depending on the presence of thef17AeandcdtIIIgenes. This partition was observed inE. colistrains of unrelated backgrounds, suggesting two different evolutionary paths ofcnf2-carrying IncF plasmids rather than divergent evolutions of NTEC2 clones. The driving forces for such divergent evolutions are not known, and further studies are required to clarify the selection of plasmid subtypes spreading virulence determinants inE. coli, in particular, plasmids of the IncF family.
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Pérez-Vázquez, María, Pedro J. Sola Campoy, Adriana Ortega, et al. "Emergence of NDM-producing Klebsiella pneumoniae and Escherichia coli in Spain: phylogeny, resistome, virulence and plasmids encoding blaNDM-like genes as determined by WGS." Journal of Antimicrobial Chemotherapy 74, no. 12 (2019): 3489–96. http://dx.doi.org/10.1093/jac/dkz366.
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Abstract Objectives NDM carbapenemases have spread worldwide. However, little information exists about the impact of NDM-producing Enterobacteriaceae in Spain. By WGS, we sought to elucidate the population structure of NDM-like-producing Klebsiella pneumoniae and Escherichia coli in Spain and to determine the plasmids harbouring blaNDM-like genes. Methods High-resolution SNP typing, core-genome MLST and plasmid reconstruction (PlasmidID) were performed on 59 NDM-like-producing K. pneumoniae and 8 NDM-like-producing E. coli isolated over an 8 year period in Spain. Results Five major epidemic clones of NDM-producing K. pneumoniae caused five important nationwide outbreaks: ST437/NDM-7, ST437/NDM-1, ST147/NDM-1, ST11/NDM-1 and ST101/NDM-1; in contrast, the spread of NDM-producing E. coli was polyclonal. Three blaNDM types were identified: blaNDM-1, 61.2%; blaNDM-7, 32.8%; and blaNDM-5, 6%. Five K. pneumoniae isolates co-produced other carbapenemases (three blaOXA-48 and two blaVIM-1). The average number of acquired resistance genes was higher in K. pneumoniae than in E. coli. The plasmids encoding blaNDM-like genes belonged to IncFII, IncFIB, IncX3, IncR, IncN and IncC types, of which IncF, IncR and IncC were associated with MDR. The genetic surroundings of blaNDM-like genes showed a highly variable region upstream of ISAba125. Conclusions In recent years NDM-producing K. pneumoniae and E. coli have emerged in Spain; the spread of a few high-risk K. pneumoniae clones such as ST437/NDM-7, ST437/NDM-1, ST147/NDM-1, ST11/NDM-1 and ST101/NDM-1 have caused several interregional outbreaks. In contrast, the spread of NDM-producing E. coli has been polyclonal. Plasmid types IncFII, IncFIB, IncX3, IncR, IncN and IncC carried blaNDM, and the same IncX3 plasmid was detected in K. pneumoniae and E. coli.
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Partridge, Sally R., Zhiyong Zong, and Jonathan R. Iredell. "Recombination in IS26and Tn2in the Evolution of Multiresistance Regions CarryingblaCTX-M-15on Conjugative IncF Plasmids from Escherichia coli." Antimicrobial Agents and Chemotherapy 55, no. 11 (2011): 4971–78. http://dx.doi.org/10.1128/aac.00025-11.
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ABSTRACTCTX-M-15 now appears to be the dominant extended-spectrum β-lactamase worldwide, and a number of different factors may contribute to this success. These include associations betweenblaCTX-M-15and particular plasmids (IncF) and/or strains, such asEscherichia coliST131, as well as the genetic contexts in which this gene is found. We previously identifiedblaCTX-M-15as the dominant ESBL gene in the western Sydney area, Australia, and found that it was carried mainly on IncF or IncI1 plasmids. Here, we have mapped the multiresistance regions of the 11 conjugative plasmids with one or more IncF replicons obtained from that survey and conducted a limited comparison of plasmid backbones. Two plasmids with only an IncFII replicon appear to be very similar to the published plasmids pC15-1a and pEK516. The remaining nine plasmids, with multiple IncF replicons, have multiresistance regions related to those of pC15-1a and pEK516, but eight contain additional modules previously found in resistance plasmids from different geographic locations that carry a variety of different resistance genes. Differences between the multiresistance regions are largely due to IS26-mediated deletions, insertions, and/or rearrangements, which can explain the observed variable associations betweenblaCTX-M-15and certain other resistance genes. We found no evidence of independent movement ofblaCTX-M-15or of a large multiresistance region between different plasmid backbones. Instead, homologous recombination between common components, such as IS26and Tn2, appeared to be more important in creating new multiresistance regions, and this may be coupled with recombination in plasmid backbones to reassort multiple IncF replicons as well as components of multiresistance regions.
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Rayamajhi, Nabin, Seung Bin Cha, Seung Won Shin, Byeong Yeal Jung, Suk-Kyung Lim, and Han Sang Yoo. "Plasmid Typing and Resistance Profiling of Escherichia fergusonii and Other Enterobacteriaceae Isolates from South Korean Farm Animals." Applied and Environmental Microbiology 77, no. 9 (2011): 3163–66. http://dx.doi.org/10.1128/aem.02188-10.
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ABSTRACTIn this study, we focused on determining the distribution and prevalence of major plasmid replicons in β-lactam-resistantEscherichia fergusoniiandEnterobacteriaceaeof animal and human origin. A high degree of plasmid variability and multiple plasmid replicons were observed among the isolates. The IncF and IncI1 replicons were the most prevalent inE. fergusoniiandSalmonella entericaserovar Indiana isolated from swine and poultry in South Korea, respectively. The presence of broad-host-range plasmid replicons such as IncN, IncA/C, IncHI1, and IncHI2 that are associated with important virulence genes and toxins as well as antimicrobial resistance determinants indicates thatE. fergusoniihas the potential to become an important pig pathogen and possible emerging opportunistic zoonotic pathogen.
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Kalaycı Yüksek, F., D. Gümüş, AC Macunluoğlu, E. Eroğlu, D. Camadan, and M. Anğ Küçüker. "Mobile resistance determinants, plasmid replicon types and phylogeny among Escherichia coli strains isolated from cats and dogs." Journal of the Hellenic Veterinary Medical Society 73, no. 4 (2023): 5039–52. http://dx.doi.org/10.12681/jhvms.30148.
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Abstract
 Multidrug resistance is a great challenge for the treatment of infectious diseases. We determined antibiotic resistance patterns, integrons, plasmid-mediated ESBL-, AmpC beta-lactamase-, carbapenemase-, colistin resistance genes, plasmid replicon types and phylogeny of fecal E. coli strains isolated from domestic cats and dogs in Turkey.
 A total of 104 fecal samples of healthy 49 cats and 55 dogs were examined. The integrons, plasmid-mediated resistance genes, plasmid replicon types and phylogroups were determined by PCR. Antimicrobial susceptibilities were performed by disc diffusion and microdilution methods.
 
 coli strains were mostly resistant to AMP (56.73%), SXT (39.42%), CTX (38.46%) and CIP (30.77%). Colistin resistance was not detected. ESBL and carbapenemase rates were 35.5 % and 7.69%, respectively. Eighty (76.9%) and 49 (47.1%) strains were harboring class I and class II integrons, respectively. Besides 12 strains were shown to possess class III integrons. The most frequently detected genes were blaCTX-M (48.08%), blaTEM (45.19%) and blaVIM (20.19%). In our study, none of strains were positive for mcr-1 and mcr-2 genes. Integrons were mostly found on plasmids of incompatibility groups IncF (71.25%) and strains bearing CTX-M and TEM carried a wide range of plasmid replicons of which IncF, IncFIB, IncK, and IncN. The majority of the strains were grouped in B2 (31.73%) and B1 (22.12%) and resistant bacteria mostly belonged to phylogroup B2.
 
 We showed an increasing trend in ESBL-producing E. coli among fecal microbiota members. E. coli strains with different plasmid replicon types and phylogroups isolated from cats and dogs can be resistant to various antibiotics which are used in human and veterinary medicine.
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Quiñones, Beatriz, Bertram G. Lee, Ashley Avilés Noriega, and Lisa Gorski. "Plasmidome of Salmonella enterica serovar Infantis recovered from surface waters in a major agricultural region for leafy greens in California." PLOS ONE 19, no. 12 (2024): e0316466. https://doi.org/10.1371/journal.pone.0316466.
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Non-typhoidal Salmonella enterica is a leading cause of gastrointestinal illnesses in the United States. Among the 2,600 different S. enterica serovars, Infantis has been significantly linked to human illnesses and is frequently recovered from broilers and chicken parts in the U.S. A key virulence determinant in serovar Infantis is the presence of the megaplasmid pESI, conferring multidrug resistance. To further characterize the virulence potential of this serovar, the present study identified the types of plasmids harbored by Infantis strains, recovered from surface waters adjacent to leafy greens farms in California. Sequencing analysis showed that each of the examined 12 Infantis strains had a large plasmid ranging in size from 78 kb to 125 kb. In addition, a second 4-kb plasmid was detected in two strains. Plasmid nucleotide queries did not identify the emerging megaplasmid pESI in the examined Infantis strains; however, the detected plasmids each had similarity to a plasmid sequence already cataloged in the nucleotide databases. Subsequent comparative analyses, based on gene presence or absence, divided the detected plasmids into five distinct clusters, and the phylogram revealed these Infantis plasmids were clustered based either on the plasmid conjugation system, IncI and IncF, or on the presence of plasmid phage genes. Assignment of the putative genes to functional categories revealed that the large plasmids contained genes implicated in cell cycle control and division, replication and recombination and defense mechanisms. Further analysis of the mobilome, including prophages and transposons, demonstrated the presence of genes implicated in the release of the bactericidal peptide microcin in the IncF plasmids and identified a Tn10 transposon conferring tetracycline resistance in one of the IncI1 plasmids. These findings indicated that the plasmids in the environmental S. enterica serovar Infantis strains from surface waters harbored a wide variety of genes associated with adaptation, survivability and antimicrobial resistance.
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Poirel, Laurent, Rémy A. Bonnin, and Patrice Nordmann. "Analysis of the Resistome of a Multidrug-Resistant NDM-1-Producing Escherichia coli Strain by High-Throughput Genome Sequencing." Antimicrobial Agents and Chemotherapy 55, no. 9 (2011): 4224–29. http://dx.doi.org/10.1128/aac.00165-11.
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ABSTRACTThe resistome of the multidrug-resistantEscherichia colistrain 271 carrying the plasmid-mediatedblaNDM-1carbapenemase gene was analyzed by high-throughput genome sequencing. The p271A plasmid carrying theblaNDM-1gene was 35.9 kb in size and possessed an IncN-type backbone that harbored a novel replicase gene. Acquisition of theblaNDM-1gene on plasmid p271A had been likely the result of a cointegration event involving the transposase of Tn5403. The expression ofblaNDM-1was associated with the insertion sequence ISAba125likely originating fromAcinetobacter baumannii. E. coli271 accumulated multiple resistance determinants, including five β-lactamase genes (comprising the extended-spectrum β-lactamase CTX-M-15), two 16S RNA methylase ArmA- and RmtB-encoding genes, and theqepAgene encoding an efflux pump involved in resistance to fluoroquinolones. These resistance genes were located on three additional plasmids, of 160 kb (IncA/C), 130 kb (IncF), and 110 kb (IncI1). In addition, several chromosomally encoded resistance determinants were identified, such as topoisomerase mutations, porin modifications and truncations, and the intrinsicampCgene ofE. colithat was weakly expressed. The multidrug resistance pattern observed forE. coli271 was therefore the result of combined chromosome- and plasmid-encoded mechanisms.
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May, Thithiwat, and Satoshi Okabe. "Escherichia coli Harboring a Natural IncF Conjugative F Plasmid Develops Complex Mature Biofilms by Stimulating Synthesis of Colanic Acid and Curli." Journal of Bacteriology 190, no. 22 (2008): 7479–90. http://dx.doi.org/10.1128/jb.00823-08.
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ABSTRACT It has been shown that Escherichia coli harboring the derepressed IncFI and IncFII conjugative F plasmids form complex mature biofilms by using their F-pilus connections, whereas a plasmid-free strain forms only patchy biofilms. Therefore, in this study we investigated the contribution of a natural IncF conjugative F plasmid to the formation of E. coli biofilms. Unlike the presence of a derepressed F plasmid, the presence of a natural IncF F plasmid promoted biofilm formation by generating the cell-to-cell mating F pili between pairs of F+ cells (approximately two to four pili per cell) and by stimulating the formation of colanic acid and curli meshwork. Formation of colanic acid and curli was required after the initial deposition of F-pilus connections to generate a three-dimensional mushroom-type biofilm. In addition, we demonstrated that the conjugative factor of F plasmid, rather than a pilus synthesis function, was involved in curli production during biofilm formation, which promoted cell-surface interactions. Curli played an important role in the maturation process. Microarray experiments were performed to identify the genes involved in curli biosynthesis and regulation. The results suggested that a natural F plasmid was more likely an external activator that indirectly promoted curli production via bacterial regulatory systems (the EnvZ/OmpR two-component regulators and the RpoS and HN-S global regulators). These data provided new insights into the role of a natural F plasmid during the development of E. coli biofilms.
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Willetts, Neil, and John Maule. "Specificities of IncF plasmid conjugation genes." Genetical Research 47, no. 1 (1986): 1–11. http://dx.doi.org/10.1017/s0016672300024447.
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SummaryThe conjugation regions of IncF plasmids are closely related in that they share extensive DNA homology, and that they specify related pili. Variations between individual conjugation gene products of different IncF plasmids have, however, been noted. We have extended these observations by carrying out a systematic survey of twelve such plasmids, to examine the numbers and the groupings of the plasmid-specific alleles of several genes required for conjugation and its control.Using vector plasmids carrying cloned origins of transfer (oriT), four different specificities were recognized, and these were correlated with the specificities of the genes with products that may act at this site (traM, traYandtraZ). ThetraYgene is the first gene of the major transfer operon, and is therefore located close to the site at which thetraJprotein acts to induce expression of the operon: correspondingly, correlation was observed between theoriT/traMYZandtraJspecificities in most of the plasmids. In turn,traJis negatively regulated by thefinOandfinPproducts acting in concert: thefinOproduct was relatively non-specific, but sixfinPalleles were identified, again with specificities correlated with those oftraJ. Our explanation for this unexpectedly large number offinPalleles derives from the concept that thefinPproduct is an RNA molecule rather than a protein. Although the conjugative pili encoded by IncF plasmids are closely related, they confer different efficiencies of plating of the various F-specific bacteriophages. We distinguished four groups on this basis, presumably resulting from differences in the primary amino-acid sequences of the pilin proteins. These groups could be related to the surface exclusion system specificities, consistent with the hypothesis that surface exclusion acts at least in part by preventing interaction between the pilus and the recipient cell surface.From these data, information about the evolutionary relationships between the twelve IncF plasmids can be deduced.
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Poirel, Laurent, Laurent Dortet, Sandrine Bernabeu, and Patrice Nordmann. "Genetic Features ofblaNDM-1-Positive Enterobacteriaceae." Antimicrobial Agents and Chemotherapy 55, no. 11 (2011): 5403–7. http://dx.doi.org/10.1128/aac.00585-11.
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ABSTRACTGenetic features associated with theblaNDM-1gene were investigated in 6Escherichia coli, 7Klebsiella pneumoniae, 1Citrobacter freundii, 1Proteus mirabilis, and 1Providencia stuartiiisolate of worldwide origin. Clonal diversity was observed for bothE. coliandK. pneumoniae. TheblaNDM-1gene was carried by different plasmid types (IncA/C, IncF, IncL/M, or untypeable) and was likely chromosome borne in two isolates. TheblaNDM-1plasmids coharbored a variety of resistance determinants, including β-lactamase genes, quinolone resistance genes, and 16S RNA methylase genes.
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Ammar, Ahmed M., Norhan K. Abd El-Aziz, Mohamed G. Aggour, et al. "A Newly Incompatibility F Replicon Allele (FIB81) in Extensively Drug-Resistant Escherichia coli Isolated from Diseased Broilers." International Journal of Molecular Sciences 25, no. 15 (2024): 8347. http://dx.doi.org/10.3390/ijms25158347.
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Multiple drug resistance (MDR) has gained pronounced attention among Enterobacterales. The transfer of multiple antimicrobial resistance genes, frequently carried on conjugative incompatibility F (IncF) plasmids and facilitating interspecies resistance transmission, has been linked to Salmonella spp. and E. coli in broilers. In Egypt, the growing resistance is exacerbated by the limited clinical efficacy of many antimicrobials. In this study, IncF groups were screened and characterized in drug-resistant Salmonella spp. and E. coli isolated from broilers. The antimicrobial resistance profile, PCR-based replicon typing of bacterial isolates pre- and post-plasmid curing, and IncF replicon allele sequence typing were investigated. Five isolates of E. coli (5/31; 16.13%) and Salmonella spp. (5/36; 13.89%) were pan-susceptible to the examined antimicrobial agents, and 85.07% of tested isolates were MDR and extensively drug-resistant (XDR). Twelve MDR and XDR E. coli and Salmonella spp. isolates were examined for the existence of IncF replicons (FII, FIA, and FIB). They shared resistance to ampicillin, ampicillin/sulbactam, amoxicillin/clavulanate, doxycycline, cefotaxime, and colistin. All isolates carried from one to two IncF replicons. The FII-FIA-FIB+ and FII-FIA+FIB- were the predominant replicon patterns. FIB was the most frequently detected replicon after plasmid curing. Three XDR E. coli isolates that were resistant to 12–14 antimicrobials carried a newly FIB replicon allele with four nucleotide substitutions: C99→A, G112→T, C113→T, and G114→A. These findings suggest that broilers are a significant reservoir of IncF replicons with highly divergent IncF-FIB plasmid incompatibility groups circulating among XDR Enterobacterales. Supporting these data with additional comprehensive epidemiological studies involving replicons other than the IncF can provide insights for implementing efficient policies to prevent the spreading of new replicons to humans.
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Moussa, Jennifer, Balig Panossian, Elie Nassour, Tamara Salloum, Edmond Abboud, and Sima Tokajian. "Detailed characterization of an IncFII plasmid carrying blaOXA-48 from Lebanon." Journal of Antimicrobial Chemotherapy 75, no. 9 (2020): 2462–65. http://dx.doi.org/10.1093/jac/dkaa181.
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Abstract Background The spread of carbapenem-resistant Enterobacteriaceae is an important challenge and an increasing healthcare problem. OXA-48 is a class D carbapenemase that is usually localized on a conjugative plasmid belonging to the IncL incompatibility group. Methods In this study, we used a combination of short- and long-read WGS approaches and molecular typing techniques to characterize the genetic environment of the smallest reported 27 029 bp IncFII plasmid carrying blaOXA-48 (pLAU-OXA48). Results The plasmid recovered from a clinical Escherichia coli isolate was positive for blaOXA-48, which was located within the Tn6237 composite transposon. Primers targeting junctions between the IncF fragment and Tn6237 for the rapid identification of pLAU-OXA48-like plasmids were designed. Conclusions To our knowledge, this is the first report showing the complete sequence of an IncFII plasmid carrying blaOXA-48 within Tn6237 using hybrid assembly of long- and short-read sequencing.
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Johnson, Timothy J., Kylie E. Siek, Sara J. Johnson, and Lisa K. Nolan. "DNA Sequence and Comparative Genomics of pAPEC-O2-R, an Avian Pathogenic Escherichia coli Transmissible R Plasmid." Antimicrobial Agents and Chemotherapy 49, no. 11 (2005): 4681–88. http://dx.doi.org/10.1128/aac.49.11.4681-4688.2005.
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ABSTRACT In this study, a 101-kb IncF plasmid from an avian pathogenic Escherichia coli (APEC) strain (APEC O2) was sequenced and analyzed, providing the first completed APEC plasmid sequence. This plasmid, pAPEC-O2-R, has functional transfer and antimicrobial resistance-encoding regions. The resistance-encoding region encodes resistance to eight groups of antimicrobial agents, including silver and other heavy metals, quaternary ammonium compounds, tetracycline, sulfonamides, aminoglycosides, trimethoprim, and beta-lactam antimicrobial agents. This region of the plasmid is unique among previously described IncF plasmids in that it possesses a class 1 integron that harbors three gene cassettes and a heavy metal resistance operon. This region spans 33 kb and is flanked by the RepFII plasmid replicon and an assortment of plasmid maintenance genes. pAPEC-O2-R also contains a 32-kb transfer region that is nearly identical to that found in the E. coli F plasmid, rendering it transferable by conjugation to plasmid-less strains of bacteria, including an APEC strain, a fecal E. coli strain from an apparently healthy bird, a Salmonella enterica serovar Typhimurium strain, and a uropathogenic E. coli strain from humans. Differences in the G+C contents of individual open reading frames suggest that various regions of pAPEC-O2-R had dissimilar origins. The presence of pAPEC-O2-R-like plasmids that encode resistance to multiple antimicrobial agents and that are readily transmissible from APEC to other bacteria suggests the possibility that such plasmids may serve as a reservoir of resistance genes for other bacteria of animal and human health significance.
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Hammerl, Jens A., Iris Klein, Erich Lanka, Bernd Appel, and Stefan Hertwig. "Genetic and Functional Properties of the Self-Transmissible Yersinia enterocolitica Plasmid pYE854, Which Mobilizes the Virulence Plasmid pYV." Journal of Bacteriology 190, no. 3 (2007): 991–1010. http://dx.doi.org/10.1128/jb.01467-07.
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ABSTRACT Yersinia strains frequently harbor plasmids, of which the virulence plasmid pYV, indigenous in pathogenic strains, has been thoroughly characterized during the last decades. Yet, it has been unknown whether the nonconjugative pYV can be transferred by helper plasmids naturally occurring in this genus. We have isolated the conjugative plasmids pYE854 (95.5 kb) and pYE966 (70 kb) from a nonpathogenic and a pathogenic Yersinia enterocolitica strain, respectively, and demonstrate that both plasmids are able to mobilize pYV. The complete sequence of pYE854 has been determined. The transfer proteins and oriT of the plasmid reveal similarities to the F factor. However, the pYE854 replicon does not belong to the IncF group and is more closely related to a plasmid of gram-positive bacteria. Plasmid pYE966 is very similar to pYE854 but lacks two DNA regions of the larger plasmid that are dispensable for conjugation.
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Liang, Huixian, Xinhui Li, and He Yan. "Identification of a Novel IncHI1B Plasmid in MDR Klebsiella pneumoniae 200 from Swine in China." Antibiotics 11, no. 9 (2022): 1225. http://dx.doi.org/10.3390/antibiotics11091225.
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Multidrug-resistant (MDR) Klebsiella pneumoniae poses a seriously threat to public health. The aim of this study was to better understand the genetic structure of its plasmids and chromosomes. The whole-genome sequence of K. pneumoniae 200 isolated from the liver of a swine with diarrhea in China was determined using PacBio RS II and Illumina MiSeq sequencing. The complete sequences of the chromosomal DNA and the plasmids were analyzed for the presence of resistance genes. The phylogenetic trees revealed that K. pneumoniae 200 displayed the closest relationship to a human-associated K. pneumoniae strain from Thailand. K. pneumoniae 200 contained two plasmids, pYhe2001 and pYhe2002, belonging to the incompatibility groups IncH-HI1B and IncF-FIA. The plasmid pYhe2001 was a novel plasmid containing four types of heavy metal resistance genes and a novel Tn6897 transposon flanked by two copies of IS26 at both ends. Mixed plasmids could be transferred from K. pneumoniae 200 to Escherichia coli DH5α through transformation together. This study reported the first time a novel plasmid pYhe2001 from swine origin K. pneumoniae 200, suggesting that the plasmids may act as reservoirs for various antimicrobial resistance genes and transport multiple resistance genes in K. pneumoniae of both animal and human origin.
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Zong, Zhiyong. "Complete Sequence of pJIE186-2, a Plasmid Carrying Multiple Virulence Factors from a Sequence Type 131 Escherichia coli O25 Strain." Antimicrobial Agents and Chemotherapy 57, no. 1 (2012): 597–600. http://dx.doi.org/10.1128/aac.01081-12.
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ABSTRACTThe complete sequence of a 137-kb plasmid, pJIE186-2, from a sequence type 131 (ST131)Escherichia colistrain was determined. pJIE186-2 contained IncF replicons (FIB, FIIA, and FIAΔ), an incomplete conjugative region, and multiple virulence factors (sitABCD,iucABCD-iutA,iroCDEN,etsABC,hlyF,iss,ompT, andvagCD) but no antimicrobial resistance genes. The host strain also had another plasmid, pJIE186-1, carrying multiple resistance genes. The two plasmids conferred selective advantages for the host strain, contributing to the recent emergence of ST131E. coli.
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Raro, Otávio Hallal Ferreira, Laurent Poirel, and Patrice Nordmann. "Effect of Zinc Oxide and Copper Sulfate on Antibiotic Resistance Plasmid Transfer in Escherichia coli." Microorganisms 11, no. 12 (2023): 2880. http://dx.doi.org/10.3390/microorganisms11122880.
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Heavy metals such as zinc (Zn) and copper (Cu) may be associated with antibiotic resistance dissemination. Our aim was to investigate whether sub-lethal dosage of Zn and Cu may enhance plasmid transfer and subsequently resistance genes dissemination. Plasmid conjugation frequencies (PCF) were performed with Escherichia coli strains bearing IncL-blaOXA-48, IncA/C-blaCMY-2, IncI1-blaCTX-M-1, IncF-blaCTX-M-1, and IncX3-blaNDM-5 as donors. Mating-out assays were performed with sub-dosages of zinc oxide (ZnO) and Cu sulfate (CuSO4). Quantification of the SOS response-associated gene expression levels and of the production of reactive oxygen species were determined. Increased PCF was observed for IncL, IncA/C, and IncX3 when treated with ZnO. PCF was only increased for IncL when treated with CuSO4. The ROS production presented an overall positive correlation with PCF after treatment with ZnO for IncL, IncA/C, and IncX3. For CuSO4 treatment, the same was observed only for IncL. No increase was observed for expression of SOS response-associated genes under CuSO4 treatment, and under ZnO treatment, we observed an increase in SOS response-associated genes only for IncX3. Our data showed that sub-dosages of ZnO and CuSO4 could significantly enhance PCF in E. coli, with a more marked effect observed with IncL, IncA/C, and IncX3 scaffolds. Our study suggested that use of certain heavy metals is not the panacea for avoiding use of antibiotics in order to prevent the dissemination of antibiotic resistance.
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Rocha-Gracia, Rosa del Carmen, Patricia Lozano-Zarain, Zita Gutiérrez Cázarez, et al. "IncFIB plasmids carrying the resistance gene blaCTX-M-15 in ESBL-producing Escherichia coli clones from pediatric patients." Journal of Infection in Developing Countries 16, no. 03 (2022): 500–506. http://dx.doi.org/10.3855/jidc.15080.
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Introduction: The emergence of extended-spectrum β-lactamases (ESBLs)-producing Escherichia coli clones are a public health concern worldwide. Scarce information does exist about the spread of ESBLs-producing E. coli in pediatric patients from developing countries.
 Methodology: E. coli strains were analyzed by multilocus-sequence-typing, pulsed-field-gel-electrophoresis and phylogenetic group. The antimicrobial-resistance genes were detected by PCR, and plasmid content by the PCR-based replicon-typing. Horizontal transfer was tested by conjugation and the location of the blaCTX-M-15 gene by Southern blot hybridization.
 Results: Thirty-two cefotaxime-resistant E. coli were recovered. Eleven of them were ESBL-producing isolates, which were well characterized and ascribed to seven sequence types and five phylogroups. The ESBL CTX-M-15 was the most prevalent enzyme (9 of 11). Plasmids of variable sizes (40-220 kb) were visualized, and the incompatibility (Inc) group FIB plasmid-replicon was detected in the ESBL strains and transferred by conjugation in 45.45% of them.
 Plasmid-borne toxin-antitoxin systems were the most frequently detected systems, strongly associated to IncF plasmids. The CTX-M-15-encoding gene was located on IncFIB plasmids.
 Conclusions: Even though a small number of ESBL-producing strains was recovered, we evidenced that IncFIB plasmids carry the blaCTX-M-15 gene, highlighting the role of IncF-type plasmids in facilitating the spread and maintenance of ESBL-encoding genes, which further favors the rapid increase of the antimicrobial resistance dissemination in disease-causing E. coli strains in pediatric patients.
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Lawley, Trevor D., Matthew W. Gilmour, James E. Gunton, Leah J. Standeven, and Diane E. Taylor. "Functional and Mutational Analysis of Conjugative Transfer Region 1 (Tra1) from the IncHI1 Plasmid R27." Journal of Bacteriology 184, no. 8 (2002): 2173–80. http://dx.doi.org/10.1128/jb.184.8.2173-2180.2002.
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ABSTRACT The conjugative transfer region 1 (Tra1) of the IncHI1 plasmid R27 was subjected to DNA sequence analysis, mutagenesis, genetic complementation, and an H-pilus-specific phage assay. Analysis of the nucleotide sequence indicated that the Tra1 region contains genes coding for mating pair formation (Mpf) and DNA transfer replication (Dtr) and a coupling protein. Insertional disruptions of 9 of the 14 open reading frames (ORFs) in the Tra1 region resulted in a transfer-deficient phenotype. Conjugative transfer was restored for each transfer mutant by genetic complementation. An intergenic region between traH and trhR was cloned and mobilized by R27, indicating the presence of an origin of transfer (oriT). The five ORFs immediately downstream of the oriT region are involved in H-pilus production, as determined by an H-pilus-specific phage assay. Three of these ORFs encode proteins homologous to Mpf proteins from IncF plasmids. Upstream of the oriT region are four ORFs required for plasmid transfer but not H-pilus production. TraI contains sequence motifs that are characteristic of relaxases from the IncP lineage but share no overall homology to known relaxases. TraJ contains both an Arc repressor motif and a leucine zipper motif. A putative coupling protein, TraG, shares a low level of homology to the TraG family of coupling proteins and contains motifs that are important for DNA transfer. This analysis indicates that the Mpf components of R27 share a common lineage with those of the IncF transfer system, whereas the relaxase of R27 is ancestrally related to that of the IncP transfer system.
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Seddon, Chloe, Gad Frankel, and Konstantinos Beis. "Structure of the outer membrane porin OmpW from the pervasive pathogen Klebsiella pneumoniae." Acta Crystallographica Section F Structural Biology Communications 80, no. 1 (2024): 22–27. http://dx.doi.org/10.1107/s2053230x23010579.
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Conjugation is the process by which plasmids, including those that carry antibiotic-resistance genes, are mobilized from one bacterium (the donor) to another (the recipient). The conjugation efficiency of IncF-like plasmids relies on the formation of mating-pair stabilization via intimate interactions between outer membrane proteins on the donor (a plasmid-encoded TraN isoform) and recipient bacteria. Conjugation of the R100-1 plasmid into Escherichia coli and Klebsiella pneumoniae (KP) recipients relies on pairing between the plasmid-encoded TraNα in the donor and OmpW in the recipient. Here, the crystal structure of K. pneumoniae OmpW (OmpWKP) is reported at 3.2 Å resolution. OmpWKP forms an eight-stranded β-barrel flanked by extracellular loops. The structures of E. coli OmpW (OmpWEC) and OmpWKP show high conservation despite sequence variability in the extracellular loops.
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Schink, Anne-Kathrin, Kristina Kadlec, and Stefan Schwarz. "Analysis ofblaCTX-M-Carrying Plasmids from Escherichia coli Isolates Collected in the BfT-GermVet Study." Applied and Environmental Microbiology 77, no. 20 (2011): 7142–46. http://dx.doi.org/10.1128/aem.00559-11.
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ABSTRACTIn this study, 417Escherichia coliisolates from defined disease conditions of companion and farm animals collected in the BfT-GermVet study were investigated for the presence of extended-spectrum β-lactamase (ESBL) genes. Three ESBL-producingE. coliisolates were identified among the 100 ampicillin-resistant isolates. TheE. coliisolates 168 and 246, of canine and porcine origins, respectively, harboredblaCTX-M-1, and the canine isolate 913 harboredblaCTX-M-15, as confirmed by PCR and sequence analysis. The isolates 168 and 246 belonged to the novel multilocus sequence typing (MLST) types ST1576 and ST1153, respectively, while isolate 913 had the MLST type ST410. The ESBL genes were located on structurally related IncN plasmids in isolates 168 and 246 and on an IncF plasmid in isolate 913. TheblaCTX-M-1upstream regions of plasmids pCTX168 and pCTX246 were similar, whereas the downstream regions showed structural differences. The genetic environment of theblaCTX-M-15gene on plasmid pCTX913 differed distinctly from that of bothblaCTX-M-1genes. Detailed sequence analysis showed that the integration of insertion sequences, as well as interplasmid recombination events, accounted for the structural variability in theblaCTX-Mgene regions.
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Höppner, Christoph, Zhenying Liu, Natalie Domke, Andrew N. Binns, and Christian Baron. "VirB1 Orthologs from Brucella suis and pKM101 Complement Defects of the Lytic Transglycosylase Required for Efficient Type IV Secretion from Agrobacterium tumefaciens." Journal of Bacteriology 186, no. 5 (2004): 1415–22. http://dx.doi.org/10.1128/jb.186.5.1415-1422.2004.
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ABSTRACT Type IV secretion systems mediate conjugative plasmid transfer as well as the translocation of virulence factors from various gram-negative pathogens to eukaryotic host cells. The translocation apparatus consists of 9 to 12 components, and the components from different organisms are believed to have similar functions. However, orthologs to proteins of the prototypical type IV system, VirB of Agrobacterium tumefaciens, typically share only 15 to 30% identical amino acids, and functional complementation between components of different type IV secretion systems has not been achieved. We here report a heterologous complementation in the case of A. tumefaciens virB1 defects with its orthologs from Brucella suis (VirB1s) and the IncN plasmid pKM101 (TraL). In contrast, expression of the genes encoding the VirB1 orthologs from the IncF plasmid (open reading frame 169) and from the Helicobacter pylori cag pathogenicity island (HP0523) did not complement VirB1 functions. The complementation of VirB1 activity was assessed by T-pilus formation, by tumor formation on wounded plants, by IncQ plasmid transfer, and by IncQ plasmid recipient assay. Replacement of the key active-site Glu residue by Ala abolished the complementation by VirB1 from B. suis and by TraL, demonstrating that heterologous complementation requires an intact lytic transglycosylase active site. In contrast, the VirB1 active-site mutant from A. tumefaciens retained considerable residual activity in various activity assays, implying that this protein exerts additional effects during the type IV secretion process.
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Lyimo, Beatus, Joram Buza, Murugan Subbiah, et al. "IncF Plasmids Are Commonly Carried by Antibiotic ResistantEscherichia coliIsolated from Drinking Water Sources in Northern Tanzania." International Journal of Microbiology 2016 (2016): 1–7. http://dx.doi.org/10.1155/2016/3103672.
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The aim of this study was to identify the replicon types of plasmids, conjugation efficiencies, and the complement of antibiotic resistance genes for a panel of multidrug resistantE. coliisolates from surface waters in northern Tanzania. Standard membrane filtration was used to isolate anduidAPCR was used to confirm the identity of strains asE. coli. Antibiotic susceptibility was determined by breakpoint assay and plasmid conjugation was determined by filter-mating experiments. PCR and sequencing were used to identify resistance genes and PCR-based replicon typing was used to determine plasmid types. Filter mating experiments indicated conjugation efficiencies ranged from 10−1to 10−7. Over 80% of the donor cells successfully passed their resistance traits and eleven different replicon types were detected (IncI1, FIC, P, FIIA, A/C, FIB, FIA, H12, K/B B/O, and N). IncF plasmids were most commonly detected (49% of isolates), followed by types IncI1 and IncA/C. Detection of these public health-relevant conjugative plasmids and antibiotic resistant traits in Tanzanian water suggests the possible pollution of these water sources from human, livestock, and wild animal wastes and also shows the potential of these water sources in the maintenance and transmission of these resistance traits between environments, animals, and people.
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Lawley, Trevor D., Matthew W. Gilmour, James E. Gunton, Dobryan M. Tracz, and Diane E. Taylor. "Functional and Mutational Analysis of Conjugative Transfer Region 2 (Tra2) from the IncHI1 Plasmid R27." Journal of Bacteriology 185, no. 2 (2003): 581–91. http://dx.doi.org/10.1128/jb.185.2.581-591.2003.
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ABSTRACT The transfer 2 region (Tra2) of the conjugative plasmid drR27 (derepressed R27) was analyzed by PSI-BLAST, insertional mutagenesis, genetic complementation, and an H-pilus assay. Tra2 contains 11 mating-pair formation (Mpf) genes that are essential for conjugative transfer, 9 of which are essential for H-pilus production (trhA, -L, -E, -K, -B, -V, -C, -P, and -W). TrhK has similarity to secretin proteins, suggesting a mechanism by which DNA could traverse the outer membrane of donors. The remaining two Mpf genes, trhU and trhN, play an auxiliary role in H-pilus synthesis and are proposed to be involved in DNA transfer and mating-pair stabilization, respectively. Conjugative transfer abilities were restored for each mutant when complemented with the corresponding transfer gene. In addition to the essential Mpf genes, three genes, trhO, trhZ, and htdA, modulate R27 transfer frequency. Disruption of trhO and trhZ severely reduced the transfer frequencies of drR27, whereas disruption of htdA greatly increased the transfer frequency of wild-type R27 to drR27 levels. A comparison of the essential transfer genes encoded by the Tra2 and Tra1 (T. D. Lawley, M. W. Gilmour, J. E. Gunton, L. J. Standeven, and D. E. Taylor, J. Bacteriol. 184:2173-2183, 2002) of R27 to other transfer systems illustrates that the R27 conjugative transfer system is a chimera composed of IncF-like and IncP-like transfer systems. Furthermore, the Mpf/type IV secretion systems encoded by IncH and IncF transfer systems are distinct from that of the IncP transfer system. The phenotypic and ecological significance of these observations is discussed.
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Li, Jun-Jie, Caressa N. Spychala, Fupin Hu, Ji-Fang Sheng, and Yohei Doi. "Complete Nucleotide Sequences ofblaCTX-M-Harboring IncF Plasmids from Community-Associated Escherichia coli Strains in the United States." Antimicrobial Agents and Chemotherapy 59, no. 6 (2015): 3002–7. http://dx.doi.org/10.1128/aac.04772-14.
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ABSTRACTCommunity-associated infections due toEscherichia coliproducing CTX-M-type extended-spectrum β-lactamases are increasingly recognized in the United States. TheblaCTX-Mgenes are frequently carried on IncF group plasmids. In this study,blaCTX-M-15-harboring plasmids pCA14 (sequence type 131 [ST131]) and pCA28 (ST44) andblaCTX-M-14-harboring plasmid pCA08 (ST131) were sequenced and characterized. The three plasmids were closely related to other IncFII plasmids from continents outside the United States in the conserved backbone region and multiresistance regions (MRRs). Each of theblaCTX-M-15-carrying plasmids pCA14 and pCA28 belonged to F31:A4:B1 (FAB [FII, FIA, FIB] formula) and showed a high level of similarity (92% coverage of pCA14 and 99% to 100% nucleotide identity), suggesting a possible common origin. TheblaCTX-M-14-carrying plasmid pCA08 belonged to F2:A2:B20 and was highly similar to pKF3-140 from China (88% coverage of pCA08 and 99% to 100% nucleotide identity). All three plasmids carried multiple antimicrobial resistance genes and modules associated with virulence and biochemical pathways, which likely confer selective advantages for their host strains. TheblaCTX-M-carrying IncFII-IA-IB plasmids implicated in community-associated infections in the United States shared key structural features with those identified from other continents, underscoring the global nature of this plasmid epidemic.
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Zhou, Xiujuan, Shanrong Yi, Dai Kuang, Chunlei Shi, and Chunbo Qu. "Analysis of Efflux Pump Contributions and Plasmid-Mediated Genetic Determinants in Ciprofloxacin-Resistant Salmonella." Pathogens 13, no. 12 (2024): 1126. https://doi.org/10.3390/pathogens13121126.
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This study aimed to explore the interactions among genetic determinants influencing ciprofloxacin resistance in Salmonella. Treatment with PAβN, an efflux pump inhibitor, resulted in a 4–32-fold reduction in the minimum inhibitory concentration (MIC) across all 18 ciprofloxacin-resistant Salmonella isolates. Notably, isolates without point mutations reverted from resistance to sensitivity. The efflux pump played a crucial role in resistance development, particularly in serovar Enteritidis, where PAβN treatment caused a more significant MIC reduction (16–32-fold) in five strains carrying the GyrA (Asp87Tyr) mutation, which initially exhibited high MICs (8 μg/mL). Several resistance genes were identified on transferable plasmids: oqxAB and aac(6′)-Ib-cr were associated with IncF plasmids in S. Enteritidis, IncA/C plasmids in S. Typhimurium, and IncHI2 plasmids in S. Virchow. Additionally, qnrS1 and/or qepA were carried by IncA/C plasmids in S. Thompson. Whole-genome sequencing revealed the presence of an oqxAB module integrated into the chromosomal DNA of S. Derby. Although the MICs of ciprofloxacin in transconjugants and transformants remained low (1–4 μg/mL), they exceeded the clinical breakpoint for susceptibility. These findings highlight the synergistic impact of efflux pumps and plasmid-mediated resistance mechanisms, contributing to the increasing prevalence of ciprofloxacin resistance and posing a significant threat to food safety.
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Ogbolu, David Olusoga, Oluwole Adebayo Daini, Afolabi Ogunledun, Oyebode Armstrong Terry Alli, and Mark Alexander Webber. "Dissemination of IncF plasmids carrying beta-lactamase genes in Gram-negative bacteria from Nigerian hospitals." Journal of Infection in Developing Countries 7, no. 05 (2013): 382–90. http://dx.doi.org/10.3855/jidc.2613.
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Introduction: Production of beta-lactamases is the predominant cause of resistance to beta-lactam antibiotics in Gram-negative bacteria. We investigated the diversity of plasmid-borne beta-lactamase genes and replicon type of the plasmids carrying the respective genes in Gram-negative bacteria recovered from clinical infection in Nigerian hospitals. Methodology: A total of 134 Gram-negative bacteria of 13 species were analyzed for antimicrobial susceptibility, phenotypic and genotypic detection of various beta-lactamases, and plasmid analysis, including replicon typing. Results: Of the 134 isolates, 111 (82.8%) contained beta-lactamases, while 28 (20.9%) carried extended-spectrum beta-lactamases. PCR and sequencing identified TEM-1 in 109 isolates (81.3%), SHV-1 in 33 isolates (24.6%), OXA-1 in 15 isolates (11.2%) and CTX-M enzymes (24 CTX-M-15 and 1 CTX-M-3) in 25 isolates (18.7%). Multiplex PCR showed that 6 isolates carried plasmidic AmpCs (ACT-1, DHA-1 and CMY-2); these enzymes were detected only in isolates possessing CTX-M beta-lactamases. Of 13 (76.9%) representative plasmids investigated in detail, 9 (69.2%) were self-transferable when selected by a beta-lactam and the plasmids once transferred coded for beta-lactam resistance. Replicon typing indicated IncF as the common vector encoding for beta-lactamases. Conclusions: The study showed a diversity of beta-lactamase genes disseminated by conjugative IncF plasmids in Gram-negative bacteria; TEM-1, SHV-1, OXA-1, CTX-M-15, CTX-M-3and plasmidic AmpC enzymes are in common circulation in Nigeria.
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Matsumura, Yasufumi, Masaki Yamamoto, Miki Nagao, Yutaka Ito, Shunji Takakura та Satoshi Ichiyama. "Association of Fluoroquinolone Resistance, Virulence Genes, and IncF Plasmids with Extended-Spectrum-β-Lactamase-Producing Escherichia coli Sequence Type 131 (ST131) and ST405 Clonal Groups". Antimicrobial Agents and Chemotherapy 57, № 10 (2013): 4736–42. http://dx.doi.org/10.1128/aac.00641-13.
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ABSTRACTThe global increase of extended-spectrum-β-lactamase (ESBL)-producingEscherichia coliis associated with the specific clonal group sequence type 131 (ST131). In order to understand the successful spread of ESBL-producingE. coliclonal groups, we characterized fluoroquinolone resistance determinants, virulence genotypes, and plasmid replicons of ST131 and another global clonal group, ST405. We investigated 41 ST131-O25b, 26 ST131-O16, 41 ST405, and 41 other ST (OST) ESBL-producing isolates, which were collected at seven acute care hospitals in Japan. The detection of ESBL types, fluoroquinolone resistance-associated mutations (including quinolone resistance-determining regions [QRDRs]), virulence genotypes, plasmid replicon types, and IncF replicon sequence types was performed using PCR and sequencing.blaCTX-M, specificallyblaCTX-M-14, was the most common ESBL gene type among the four groups. Ciprofloxacin resistance was found in 90% of ST131-O25b, 19% of ST131-O16, 100% of ST405, and 54% of OST isolates. Multidrug resistance was more common in the ST405 group than in the ST131-O25 group (56% versus 32%;P= 0.045). All ST131-O25b isolates except one had four characteristic mutations in QRDRs, but most of the isolates from the other three groups had three mutations in common. The ST131-O25b and ST405 groups had larger numbers of virulence genes than the OST group. All of the ST131-O25b and ST405 isolates and most of the ST131-O16 and OST isolates carried IncF replicons. The most prevalent IncF replicon sequence types differed between the four clonal groups. Both the ST131-O25b and ST405 clonal groups had a fluoroquinolone resistance mechanism in QRDRs, multidrug resistance, high virulence, and IncF plasmids, suggesting the potential for further global expansion and a need for measures against these clonal groups.
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Abbassi, Mohamed Salah, Hajer Kilani, Islem Abid, et al. "Genetic Background of Antimicrobial Resistance in Multiantimicrobial-Resistant Escherichia coli Isolates from Feces of Healthy Broiler Chickens in Tunisia." BioMed Research International 2021 (October 1, 2021): 1–7. http://dx.doi.org/10.1155/2021/1269849.
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Multiantimicrobial-resistant Escherichia coli isolates are a global human health problem causing increasing morbidity and mortality. Genes encoding antimicrobial resistance are mainly harbored on mobile genetic elements (MGEs) such as transposons and plasmids as well as integrons, which enhance their rapid spread. The aim of this study was to characterize 83 multiantimicrobial-resistant E. coli isolates recovered from healthy broiler chickens. Among 78 tetracycline-resistant isolates, the tetA, tetB, and tetC genes were detected in 59 (75.6%), 14 (17.9%), and one (1.2%) isolates, respectively. The sul1, sul2, and sul3 genes were detected 31 (46.2%), 16 (23.8%), and 6 (8.9%) isolates, respectively, among 67 sulfonamide-resistant isolates. The PCR-based replicon typing method showed plasmids in 29 isolates, IncFIB (19), IncI1-Iγ (17), IncF (14), IncK (14), IncFIC (10), IncP (8), IncY (3), IncHI2 (1), and IncX (1). The class 1 and 2 integrons were detected in 57 and 2 isolates, respectively; one isolate harbored both integrons. Seven and one gene cassette arrays were identified in class 1 and class 2 integrons, respectively. Our findings show that multiantimicrobial-resistant E. coli isolates from chickens serve as reservoirs of highly diverse and abundant tet and sul genes and plasmid replicons. Such isolates and MGEs pose a potential health threat to the public and animal farming.
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Dealtry, Simone, Peter N. Holmsgaard, Vincent Dunon, et al. "Shifts in Abundance and Diversity of Mobile Genetic Elements after the Introduction of Diverse Pesticides into an On-Farm Biopurification System over the Course of a Year." Applied and Environmental Microbiology 80, no. 13 (2014): 4012–20. http://dx.doi.org/10.1128/aem.04016-13.
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ABSTRACTBiopurification systems (BPS) are used on farms to control pollution by treating pesticide-contaminated water. It is assumed that mobile genetic elements (MGEs) carrying genes coding for enzymes involved in degradation might contribute to the degradation of pesticides. Therefore, the composition and shifts of MGEs, in particular, of IncP-1 plasmids carried by BPS bacterial communities exposed to various pesticides, were monitored over the course of an agricultural season. PCR amplification of total community DNA using primers targeting genes specific to different plasmid groups combined with Southern blot hybridization indicated a high abundance of plasmids belonging to IncP-1, IncP-7, IncP-9, IncQ, and IncW, while IncU and IncN plasmids were less abundant or not detected. Furthermore, the integrase genes of class 1 and 2 integrons (intI1,intI2) and genes encoding resistance to sulfonamides (sul1,sul2) and streptomycin (aadA) were detected and seasonality was revealed. Amplicon pyrosequencing of the IncP-1trfAgene coding for the replication initiation protein revealed high IncP-1 plasmid diversity and an increase in the abundance of IncP-1β and a decrease in the abundance of IncP-1ε over time. The data of the chemical analysis showed increasing concentrations of various pesticides over the course of the agricultural season. As an increase in the relative abundances of bacteria carrying IncP-1β plasmids also occurred, this might point to a role of these plasmids in the degradation of many different pesticides.
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Puangseree, Jiratchaya, Rangsiya Prathan, Songsak Srisanga, and Rungtip Chuanchuen. "Molecular basis of the persistence of chloramphenicol resistance among Escherichia coli and Salmonella spp. from pigs, pork and humans in Thailand." PLOS ONE 19, no. 5 (2024): e0304250. http://dx.doi.org/10.1371/journal.pone.0304250.
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This study aimed to investigate the potential mechanisms associated with the persistence of chloramphenicol (CHP) resistance in Escherichia coli and Salmonella enterica isolated from pigs, pork, and humans in Thailand. The CHP-resistant E. coli (n = 106) and Salmonella (n = 57) isolates were tested for their CHP susceptibility in the presence and absence of phenylalanine arginine β-naphthylamide (PAβN). The potential co-selection of CHP resistance was investigated through conjugation experiments. Whole genome sequencing (WGS) was performed to analyze the E. coli (E329, E333, and E290) and Salmonella (SA448, SA461, and SA515) isolates with high CHP MIC (32–256 μg/mL) and predominant plasmid replicon types. The presence of PAβN significantly reduced the CHP MICs (≥4-fold) in most E. coli (67.9%) and Salmonella (64.9%). Ampicillin, tetracycline, and streptomycin co-selected for CHP-resistant Salmonella and E. coli-transconjugants carrying cmlA. IncF plasmids were mostly detected in cmlA carrying Salmonella (IncFIIAs) and E. coli (IncFIB and IncF) transconjugants. The WGS analysis revealed that class1 integrons with cmlA1 gene cassette flanked by IS26 and TnAs1 were located on IncX1 plasmid, IncFIA(HI1)/HI1B plasmids and IncFII/FIB plasmids. IncFIA(HI1)/HI1B/Q1in SA448 contained catA flanked by IS1B and TnAs3. In conclusion, cross resistance through proton motive force-dependent mechanisms and co-selection by other antimicrobial agents involved the persistence of CHP-resistance in E. coli in this collection. Dissemination of CHP-resistance genes was potentially facilitated by mobilization via mobile genetic elements.
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Finlay, B. B., L. S. Frost, W. Paranchych, and N. S. Willetts. "Nucleotide sequences of five IncF plasmid finP alleles." Journal of Bacteriology 167, no. 2 (1986): 754–57. http://dx.doi.org/10.1128/jb.167.2.754-757.1986.
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McGann, Patrick, Erik Snesrud, Ana C. Ong, et al. "War Wound Treatment Complications Due to Transfer of an IncN Plasmid HarboringblaOXA-181from Morganella morganii to CTX-M-27-Producing Sequence Type 131 Escherichia coli." Antimicrobial Agents and Chemotherapy 59, no. 6 (2015): 3556–62. http://dx.doi.org/10.1128/aac.04442-14.
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ABSTRACTA 22-year-old male developed a recurrent sacral abscess associated with embedded shrapnel following a blast injury. Cultures grew extended-spectrum β-lactamase (ESBL)-producing, carbapenem-susceptibleEscherichia coli. Ertapenem was administered, but the infection recurred after each course of antibiotics. Initial surgical interventions were unsuccessful, and subsequent cultures yieldedE. coliandMorganella morganii, both nonsusceptible to carbapenems. The isolates were Carba NP test negative, gave ambiguous results with the modified Hodge test, and amplified theblaOXA48-like gene by real-time PCR. AllE. coliisolates were sequence type 131 (ST131), carried nine resistance genes (includingblaCTX-M-27) on an IncF plasmid, and were identical by genome sequencing, except for 150 kb of plasmid DNA in carbapenem-nonsusceptible isolates only. Sixty kilobases of this was shared byM. morganiiand represented an IncN plasmid harboringblaOXA-181. InM. morganii, the gene was flanked by IS3000and ISKpn19, but in all but one of theE. coliisolates containingblaOXA-181, a second copy of ISKpn19had inserted adjacent to IS3000. To the best of our knowledge, this is the first report ofblaOXA-181in the virulent ST131 clonal group and carried by the promiscuous IncN family of plasmids. The tendency ofM. morganiito have high MICs of imipenem, ablaOXA-181substrate profile that includes penicillins but not extended-spectrum cephalosporins, and weak carbapenemase activity almost resulted in the presence ofblaOXA-181being overlooked. We highlight the importance of surveillance for carbapenem resistance in all species, even those with intrinsic resistances, and the value of advanced molecular techniques in detecting subtle genetic changes.
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Yao, Yancheng, Can Imirzalioglu, Linda Falgenhauer, et al. "Plasmid-Mediated Spread of Carbapenem Resistance in Enterobacterales: A Three-Year Genome-Based Survey." Antibiotics 13, no. 8 (2024): 682. http://dx.doi.org/10.3390/antibiotics13080682.
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The worldwide emergence and dissemination of carbapenem-resistant Gram-negative bacteria (CRGNB) is a challenging problem of antimicrobial resistance today. Outbreaks with CRGNB have severe consequences for both the affected healthcare settings as well as the patients with infection. Thus, bloodstream infections caused by metallo-ß-lactamase-producing Enterobacterales can often have clinical implications, resulting in high mortality rates due to delays in administering effective treatment and the limited availability of treatment options. The overall threat of CRGNB is substantial because carbapenems are used to treat infections caused by ESBL-producing Enterobacterales which also exist with high frequency within the same geographical regions. A genome-based surveillance of 589 CRGNB from 61 hospitals across the federal state Hesse in Germany was implemented using next-generation sequencing (NGS) technology to obtain a high-resolution landscape of carbapenem-resistant isolates over a three-year period (2017–2019). The study examined all reportable CRGNB isolates submitted by participating hospitals. This included isolates carrying known carbapenemases (435) together with carbapenem-resistant non-carbapenemase producers (154). Predominant carbapenemase producers included Klebsiella pneumoniae, Escherichia coli, Citrobacter freundii and Acinetobacter baumannii. Over 80% of 375 carbapenem-resistant determinants including KPC-, NDM-, VIM- and OXA-48-like ones detected in 520 Enterobacterales were plasmid-encoded, and half of these were dominated by a few incompatibility (Inc) types, viz., IncN, IncL/M, IncFII and IncF(K). Our results revealed that plasmids play an extraordinary role in the dissemination of carbapenem resistance in the heterogeneous CRGNB population. The plasmids were also associated with several multispecies dissemination events and local outbreaks throughout the study period, indicating the substantial role of horizontal gene transfer in carbapenemase spread. Furthermore, due to vertical and horizontal plasmid transfer, this can have an impact on implant-associated infections and is therefore important for antibiotic-loaded bone cement and drug-containing devices in orthopedic surgery. Future genomic surveillance projects should increase their focus on plasmid characterization.
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Szmolka, Ama, Barbara Lestár, Judit Pászti, Péter Z. Fekete, and Béla Nagy. "Conjugative IncF and IncI1 plasmids with tet(A) and class 1 integron conferring multidrug resistance in F18+ porcine enterotoxigenic E. coli." Acta Veterinaria Hungarica 63, no. 4 (2015): 425–43. http://dx.doi.org/10.1556/004.2015.040.
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Enterotoxigenic E. coli (ETEC) bacteria frequently cause watery diarrhoea in newborn and weaned pigs. Plasmids carrying genes of different enterotoxins and fimbrial adhesins, as well as plasmids conferring antimicrobial resistance are of prime importance in the epidemiology and pathogenesis of ETEC. Recent studies have revealed the significance of the porcine ETEC plasmid pTC, carrying tetracycline resistance gene tet(B) with enterotoxin genes. In contrast, the role of tet(A) plasmids in transferring resistance of porcine ETEC is less understood. The objective of the present study was to provide a comparative analysis of antimicrobial resistance and virulence gene profiles of porcine post-weaning ETEC strains representing pork-producing areas in Central Europe and in the USA, with special attention to plasmids carrying the tet(A) gene. Antimicrobial resistance phenotypes and genotypes of 87 porcine ETEC strains isolated from cases of post-weaning diarrhoea in Austria, the Czech Republic, Hungary and the Midwest USA was determined by disk diffusion and by PCR. Central European strains carrying tet(A) or tet(B) were further subjected to molecular characterisation of their tet plasmids. Results indicated that > 90% of the ETEC strains shared a common multidrug resistant (MDR) pattern of sulphamethoxazole (91%), tetracycline (84%) and streptomycin (80%) resistance. Tetracycline resistance was most frequently determined by the tet(B) gene (38%), while tet(A) was identified in 26% of all isolates with wide ranges for both tet gene types between some countries and with class 1 integrons and resistance genes co-transferred by conjugation. The virulence gene profiles included enterotoxin genes (lt, sta and/or stb), as well as adhesin genes (k88/f4, f18). Characterisation of two representative tet(A) plasmids of porcine F18+ ETEC from Central Europe revealed that the IncF plasmid (pES11732) of the Czech strain (~120 kb) carried tet(A) in association with catA1 for chloramphenicol resistance. The IncI1 plasmid (pES2172) of the Hungarian strain (~138 kb) carried tet(A) gene and a class 1 integron with an unusual variable region of 2,735 bp composed by two gene cassettes: estX-aadA1 encoding for streptothricin-spectinomycin/streptomycin resistance exemplifying simultaneous recruitment, assembly and transfer of multidrug resistance genes by the tet(A) plasmid of porcine ETEC. By this we provide the first description of IncF and IncI1 type plasmids of F18+ porcine enterotoxigenic E. coli responsible for cotransfer of the tet(A) gene with multidrug resistance. Additionally, the unusual determinant estX, encoding for streptothricin resistance, is first reported here in porcine enterotoxigenic E. coli.
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Tóth, Kinga, Ivelina Damjanova, Levente Laczkó, et al. "Genomic Epidemiology of C2/H30Rx and C1-M27 Subclades of Escherichia coli ST131 Isolates from Clinical Blood Samples in Hungary." Antibiotics 13, no. 4 (2024): 363. http://dx.doi.org/10.3390/antibiotics13040363.
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Extended-spectrum β-lactamase-producing Escherichia coli ST131 has become widespread worldwide. This study aims to characterize the virulome, resistome, and population structure of E. coli ST131 isolates from clinical blood samples in Hungary. A total of 30 C2/H30Rx and 33 C1-M27 ST131 isolates were selected for Illumina MiSeq sequencing and 30 isolates for MinION sequencing, followed by hybrid de novo assembly. Five C2/H30Rx and one C1-M27 cluster were identified. C1-M27 isolates harbored the F1:A2:B20 plasmid in 93.9% of cases. Long-read sequencing revealed that blaCTX-M-27 was on plasmids. Among the C2/H30Rx isolates, only six isolates carried the C2-associated F2:A1:B- plasmid type. Of 19 hybrid-assembled C2/H30Rx genomes, the blaCTX-M-15 gene was located on plasmid only in one isolate, while in the other isolates, ISEcp1 or IS26-mediated chromosomal integration of blaCTX-M-15 was detected in unique variations. In one isolate a part of F2:A1:B- plasmid integrated into the chromosome. These results suggest that CTX-M-15-producing C2/H30Rx and CTX-M-27-producing C1-M27 subclades may have emerged and spread in different ways in Hungary. While blaCTX-M-27 was carried mainly on the C1/H30R-associated F1:A2:B20 plasmid, the IncF-like plasmids of C2/H30Rx or its composite transposons have been incorporated into the chromosome through convergent evolutionary processes.
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Gancz, Ayala, Kira Kondratyeva, Dorit Cohen-Eli, and Shiri Navon-Venezia. "Genomics and Virulence of Klebsiella pneumoniae Kpnu95 ST1412 Harboring a Novel Incf Plasmid Encoding Blactx-M-15 and Qnrs1 Causing Community Urinary Tract Infection." Microorganisms 9, no. 5 (2021): 1022. http://dx.doi.org/10.3390/microorganisms9051022.
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The emergence of extended-spectrum β-lactamase (ESBL)-producing multidrug resistant Klebsiella pneumoniae causing community urinary tract infections (CA-UTI) in healthy women undermines effective treatment and poses a public health concern. We performed a comprehensive genomic analysis (Illumina and MinION) and virulence studies using Caenorhabditis elegans nematodes to evaluate KpnU95, a blaCTX-M-15-producing CA-UTI K. pneumoniae strain. Whole genome sequencing identified KpnU95 as sequence type 1412 and revealed the chromosomal and plasmid-encoding resistome, virulome and persistence features. KpnU95 possess a wide virulome and caused complete C. elegans killing. The strain harbored a single novel 180.3Kb IncFIB(K) plasmid (pKpnU95), which encodes ten antibiotic resistance genes, including blaCTX-M-15 and qnrS1 alongside a wide persistome encoding heavy metal and UV resistance. Plasmid curing and reconstitution were used for loss and gain studies to evaluate its role on bacterial resistance, fitness and virulence. Plasmid curing abolished the ESBL phenotype, decreased ciprofloxacin MIC and improved bacterial fitness in artificial urine accompanied with enhanced copper tolerance, without affecting bacterial virulence. Meta-analysis supported the uniqueness of pKpnU95 and revealed plasmid-ST1412 lineage adaptation. Overall, our findings provide translational data on a CA-UTI K. pneumoniae ST1412 strain and demonstrates that ESBL-encoding plasmids play key roles in multidrug resistance and in bacterial fitness and persistence.
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Feng, Yu, Lu Liu, Alan McNally, and Zhiyong Zong. "Coexistence of three blaKPC-2 genes on an IncF/IncR plasmid in ST11 Klebsiella pneumoniae." Journal of Global Antimicrobial Resistance 17 (June 2019): 90–93. http://dx.doi.org/10.1016/j.jgar.2018.11.017.
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Falodun, Olutayo Israel, Akeem Ganiyu Rabiu, Abidemi Joseph Marcus, Rotimi Ayodeji Dada, and Mobolaji Christianah Afolabi. "Characterization of virulent Escherichia coli in healthy pet dog feces: Implications for public health." Journal of Istanbul Veterinary Sciences 8, no. 1 (2024): 5–12. http://dx.doi.org/10.30704/http-www-jivs-net.1407165.
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The characterization of Escherichia coli that colonizes pets is necessary to maintain animal health and to reduce the chance of transmission to owners. In this study, we investigated the incidence of potentially virulent E. coli inhabiting healthy pet dogs as a risk of infection to pet owners. Antibiotic-resistant E. coli isolated from freshly passed dog feces were whole-genome sequenced using Illumina chemistry and classified into pathogenic lineages using pathogen-specific markers. The antimicrobial resistance genes (ARGs), virulence-associated genes (VAGs), and plasmids were respectively predicted using the ResFinder, VirulenceFinder, and PlasmidFinder. Of the 32 isolates, 13 carried resistance genes such that four, six, and 11 contained β-lactam (blaTEM), aminoglycoside [aac-6(Ib7)/ant-3(Iia)/aph-3(Ib)/aph-6(Id)] and tetracycline (tet) resistance genes, respectively. The IncF plasmids were most prevalent (n=12, 38.71%) but the highly self-conjugative IncN plasmids occurred simultaneously with the plasmid-borne [quinolones (QnrS1/QnrB7) and sulfonamide (sul3)] ARGs in ≥ 2 E. coli. One E. coli each was classified as avian pathogenic E. coli, atypical enteropathogenic E. coli, enterotoxigenic E. coli, Shiga toxin-producing enteroaggregative E. coli, and enteroaggregative E. coli. Pet feces should be carefully handled because they contain virulent and drug-resistant E. coli.
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Ho, Pak-Leung, River C. Wong, Stephanie W. Lo, Kin-Hung Chow, Samson S. Wong, and Tak-Lun Que. "Genetic identity of aminoglycoside-resistance genes in Escherichia coli isolates from human and animal sources." Journal of Medical Microbiology 59, no. 6 (2010): 702–7. http://dx.doi.org/10.1099/jmm.0.015032-0.
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A bacterial collection (n=249) obtained in Hong Kong from 2002 to 2004 was used to investigate the molecular epidemiology of aminoglycoside resistance among Escherichia coli isolates from humans and food-producing animals. Of these, 89 isolates were gentamicin-sensitive (human n=60, animal n=29) and 160 isolates were gentamicin-resistant (human n=107, animal n=53). Overall, 84.1 % (90/107) and 75.5 % (40/53) of the gentamicin-resistant isolates from human and animal sources, respectively, were found to possess the aacC2 gene. The aacC2 gene for 20 isolates (10 each for human and animal isolates) was sequenced. Two alleles were found that were equally distributed in human and animal isolates. PFGE showed that the gentamicin-resistant isolates exhibited diverse patterns with little clonality. In some isolates, the aacC2 gene was encoded on large transferable plasmids of multiple incompatibility groups (IncF, IncI1 and IncN). An IncFII plasmid of 140 kb in size was shared by one human and three animal isolates. In summary, this study showed that human and animal isolates share the same pool of resistance genes.
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Yang, Qiu E., Timothy Rutland Walsh, Bao Tao Liu, et al. "Complete Sequence of the FII Plasmid p42-2, CarryingblaCTX-M-55,oqxAB,fosA3, andfloRfrom Escherichia coli." Antimicrobial Agents and Chemotherapy 60, no. 7 (2016): 4336–38. http://dx.doi.org/10.1128/aac.00475-16.
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ABSTRACTWe sequenced a novel conjugative multidrug resistance IncF plasmid, p42-2, isolated fromEscherichia colistrain 42-2, previously identified in China. p42-2 is 106,886 bp long, composed of a typical IncFII-type backbone (∼54 kb) and one distinct acquired DNA region spanning ∼53 kb, harboring 12 antibiotic resistance genes [blaCTX-M-55,oqxA,oqxB,fosA3,floR,tetA(A),tetA(R),strA,strB,sul2,aph(3′)-II, and ΔblaTEM-1]. The spread of these multidrug resistance determinants on the same plasmid is of great concern and, because of coresistance to antibiotics from different classes, is therapeutically challenging.
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Calhau, Vera, Catarina Mendes, Angelina Pena, Nuno Mendonça, and Gabriela Jorge Da Silva. "Virulence and plasmidic resistance determinants of Escherichia coli isolated from municipal and hospital wastewater treatment plants." Journal of Water and Health 13, no. 2 (2014): 311–18. http://dx.doi.org/10.2166/wh.2014.327.
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Escherichia coli is simultaneously an indicator of water contamination and a human pathogen. This study aimed to characterize the virulence and resistance of E. coli from municipal and hospital wastewater treatment plants (WWTPs) in central Portugal. From a total of 193 isolates showing reduced susceptibility to cefotaxime and/or nalidixic acid, 20 E. coli with genetically distinct fingerprint profiles were selected and characterized. Resistance to antimicrobials was determined using the disc diffusion method. Extended spectrum β-lactamase and plasmid-mediated quinolone resistance genes, phylogroups, pathogenicity islands (PAIs) and virulence genes were screened by polymerase chain reaction (PCR). CTX-M producers were typed by multilocus sequence typing. Resistance to beta-lactams was associated with the presence of blaTEM,blaSHV, blaCTX-M-15 and blaCTX-M-32. Plasmid-mediated quinolone resistance was associated with qnrA, qnrS and aac(6′)-Ib-cr. Aminoglycoside resistance and multidrug-resistant phenotypes were also detected. PAI IV536, PAI IICFT073, PAI II536 and PAI ICFT073, and uropathogenic genes iutA, papAH and sfa/foc were detected. With regard to the clinical ST131 clone, it carried blaCTX-M-15, blaTEM-type, qnrS and aac(6′)-lb-cr; IncF and IncP plasmids, and virulence factors PAI IV536, PAI ICFT073, PAI IICFT073, iutA, sfa/foc and papAH were identified in the effluent of a hospital plant. WWTPs contribute to the dissemination of virulent and resistant bacteria in water ecosystems, constituting an environmental and public health risk.
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Palm, Martin, Alfred Fransson, Julia Hultén, et al. "The Effect of Heavy Metals on Conjugation Efficiency of an F-Plasmid in Escherichia coli." Antibiotics 11, no. 8 (2022): 1123. http://dx.doi.org/10.3390/antibiotics11081123.
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Conjugation, the process by which conjugative plasmids are transferred between bacteria, is regarded as a major contributor to the spread of antibiotic resistance, in both environmental and clinical settings. Heavy metals are known to co-select for antibiotic resistance, but the impact of the presence of these metals on conjugation itself is not clear. Here, we systematically investigate the impact that five heavy metals (arsenic, cadmium, copper, manganese, and zinc) have on the transfer of an IncF conjugative plasmid in Escherichia coli. Our results show that two of the metals, cadmium and manganese, have no significant impact, while arsenic and zinc both reduce conjugation efficiency by approximately 2-fold. Copper showed the largest impact, with an almost 100-fold decrease in conjugation efficiency. This was not mediated by any change in transcription from the major Py promoter responsible for transcription of the conjugation machinery genes. Further, we show that in order to have this severe impact on the transfer of the plasmid, copper sulfate needs to be present during the mating process, and we suggest explanations for this.
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Markovska, Rumyana, Ines Schneider, Dobrinka Ivanova, Ivan Mitov, and Adolf Bauernfeind. "Predominance of IncL/M and IncF plasmid types among CTX-M-ESBL-producingEscherichia coliandKlebsiella pneumoniaein Bulgarian hospitals." APMIS 122, no. 7 (2013): 608–15. http://dx.doi.org/10.1111/apm.12204.
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VUBIL, D., R. FIGUEIREDO, T. REIS, C. CANHA, L. BOAVENTURA, and G. J. DA SILVA. "Outbreak of KPC-3-producing ST15 and ST348Klebsiella pneumoniaein a Portuguese hospital." Epidemiology and Infection 145, no. 3 (2016): 595–99. http://dx.doi.org/10.1017/s0950268816002442.
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SUMMARYTo date, only a few sporadic cases of infections due toKlebsiella pneumoniaecarbapenemase (KPC) producers have been reported in Portugal. Here, we report for the first time an outbreak ofK. pneumoniaeKPC-3 producers in a tertiary-care hospital during 2013. Twenty-seven ertapenem-resistantK. pneumoniaewere identified in patients at a tertiary-care hospital during 2013 isolated predominantly from urine (48·1%) and blood (25·9%) cultures. All isolates were highly resistant toβ-lactam antibiotics and most showed intermediate resistance to imipenem. The more frequentβ-lactamases were TEM- (77·7%), CTX-M- (70·3%) and KPC-type (66·6%). KPC-3 was identified by sequencing. TheblaKPC−3gene was associated with an IncF plasmid, and efficiently transferred toE. coliJ53. Pulsed-field gel electrophoresis typing revealed three clusters of isolates which were further characterized by multi-locus sequence typing as ST11, ST15 and ST348. Ertapenem-resistant ST15 was already in circulation in the hospital, related to expression of OmpK36 modified porin, but the other two sequence types had not been previously found in the hospital. We conclude that the IncF plasmid mediated transfer of KPC-3 in the outbreak and that implementation of carbapenemase gene screening in isolates from patients on admission to hospital is advisable in order to control dissemination of these antimicrobial resistance elements.
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Fontana, Herrison, Brenda Cardoso, Fernanda Esposito, Aline Valerio de Lima, Jorge Luiz Mello Sampaio, and Nilton Lincopan. "Small IncQ1 Plasmid Encoding KPC-2 Expands to Invasive Non-Typhoidal Salmonella." Antimicrobial Agents and Chemotherapy, August 30, 2021. http://dx.doi.org/10.1128/aac.01552-21.
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The pandemic status of KPC-type carbapenemases has been linked especially to IncF, IncN, IncX, IncA/C, IncP, IncL/M, and IncR plasmids associated with the mobile element Tn 4401 , which have rapidly disseminated among international clones of Escherichia coli and Klebsiella complex (1, 2).…
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Mahérault, Anne-Claire, Harry Kemble, Mélanie Magnan, et al. "Advantage of the F2:A1:B- IncF Pandemic Plasmid over IncC Plasmids in In Vitro Acquisition and Evolution of blaCTX-M Gene-Bearing Plasmids in Escherichia coli." Antimicrobial Agents and Chemotherapy 63, no. 10 (2019). http://dx.doi.org/10.1128/aac.01130-19.
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ABSTRACT Despite a fitness cost imposed on bacterial hosts, large conjugative plasmids play a key role in the diffusion of resistance determinants, such as CTX-M extended-spectrum β-lactamases. Among the large conjugative plasmids, IncF plasmids are the most predominant group, and an F2:A1:B- IncF-type plasmid encoding a CTX-M-15 variant was recently described as being strongly associated with the emerging worldwide Escherichia coli sequence type 131 (ST131)-O25b:H4 H30Rx/C2 sublineage. In this context, we investigated the fitness cost of narrow-range F-type plasmids, including the F2:A1:B- IncF-type CTX-M-15 plasmid, and of broad-range C-type plasmids in the K-12-like J53-2 E. coli strain. Although all plasmids imposed a significant fitness cost to the bacterial host immediately after conjugation, we show, using an experimental-evolution approach, that a negative impact on the fitness of the host strain was maintained throughout 1,120 generations with the IncC-IncR plasmid, regardless of the presence or absence of cefotaxime, in contrast to the F2:A1:B- IncF plasmid, whose cost was alleviated. Many chromosomal and plasmid rearrangements were detected after conjugation in transconjugants carrying the IncC plasmids but not in transconjugants carrying the F2:A1:B- IncF plasmid, except for insertion sequence (IS) mobilization from the fliM gene leading to the restoration of motility of the recipient strains. Only a few mutations occurred on the chromosome of each transconjugant throughout the experimental-evolution assay. Our findings indicate that the F2:A1:B- IncF CTX-M-15 plasmid is well adapted to the E. coli strain studied, contrary to the IncC-IncR CTX-M-15 plasmid, and that such plasmid-host adaptation could participate in the evolutionary success of the CTX-M-15-producing pandemic E. coli ST131-O25b:H4 lineage.