scholarly journals Novel Tetracycline Resistance Determinant Isolated from an Environmental Strain of Serratia marcescens

2007 ◽  
Vol 73 (7) ◽  
pp. 2199-2206 ◽  
Author(s):  
Stuart A. Thompson ◽  
Elizabeth V. Maani ◽  
Angela H. Lindell ◽  
Catherine J. King ◽  
J. Vaun McArthur
Keyword(s):  
Serratia Marcescens ◽  
Efflux Pump ◽  
Tetracycline Resistance ◽  
Amino Acid Identity ◽  
Resistance Determinant ◽  
Mercury Resistance ◽  
Resistance Locus ◽  
Environmental Strain ◽  
Apparent Lack ◽  
E Coli

ABSTRACT Resistances to tetracycline and mercury were identified in an environmental strain of Serratia marcescens isolated from a stream highly contaminated with heavy metals. As a step toward addressing the mechanisms of coselection of heavy metal and antibiotic resistances, the tetracycline resistance determinant was cloned in Escherichia coli. Within the cloned 13-kb segment, the tetracycline resistance locus was localized by deletion analysis and transposon mutagenesis. DNA sequence analysis of an 8.0-kb region revealed a novel gene [tetA(41)] that was predicted to encode a tetracycline efflux pump. Phylogenetic analysis showed that the TetA(41) protein was most closely related to the Tet(39) efflux protein of Acinetobacter spp. yet had less than 80% amino acid identity with known tetracycline efflux pumps. Adjacent to the tetA(41) gene was a divergently transcribed gene [tetR(41)] predicted to encode a tetracycline-responsive repressor protein. The tetA(41)-tetR(41) intergenic region contained putative operators for TetR(41) binding. The tetA(41) and tetR(41) promoters were analyzed using lacZ fusions, which showed that the expression of both the tetA(41) and tetR(41) genes exhibited TetR(41)-dependent regulation by subinhibitory concentrations of tetracycline. The apparent lack of plasmids in this S. marcescens strain, as well as the presence of metabolic genes adjacent to the tetracycline resistance locus, suggested that the genes were located on the S. marcescens chromosome and may have been acquired by transduction. The cloned Tet 41 determinant did not confer mercury resistance to E. coli, confirming that Tet 41 is a tetracycline-specific efflux pump rather than a multidrug transporter.

scholarly journals A New Tetracycline Efflux Gene, tet(40), Is Located in Tandem with tet(O/32/O) in a Human Gut Firmicute Bacterium and in Metagenomic Library Clones

2008 ◽  
Vol 52 (11) ◽  
pp. 4001-4009 ◽  
Author(s):  
Katarzyna A. Kazimierczak ◽  
Marco T. Rincon ◽  
Andrea J. Patterson ◽  
Jennifer C. Martin ◽  
Pauline Young ◽  
...  
Keyword(s):  
Efflux Pump ◽  
Metagenomic Library ◽  
Tetracycline Resistance ◽  
Mobile Element ◽  
Amino Acid Identity ◽  
Donor Strain ◽  
Human Gut ◽  
Efflux Pump Inhibitor ◽  
E Coli ◽  
Tetracycline Resistance Genes

ABSTRACT The bacterium Clostridium saccharolyticum K10, isolated from a fecal sample obtained from a healthy donor who had received long-term tetracycline therapy, was found to carry three tetracycline resistance genes: tet(W) and the mosaic tet(O/32/O), both conferring ribosome protection-type resistance, and a novel, closely linked efflux-type resistance gene designated tet(40). tet(40) encodes a predicted membrane-associated protein with 42% amino acid identity to tetA(P). Tetracycline did not accumulate in Escherichia coli cells expressing the Tet(40) efflux protein, and resistance to tetracycline was reduced when cells were incubated with an efflux pump inhibitor. E. coli cells carrying tet(40) had a 50% inhibitory concentration of tetracycline of 60 μg/ml. Analysis of a transconjugant from a mating between donor strain C. saccharolyticum K10 and the recipient human gut commensal bacterium Roseburia inulinivorans suggested that tet(O/32/O) and tet(40) were cotransferred on a mobile element. Sequence analysis of a 37-kb insert identified on the basis of tetracycline resistance from a metagenomic fosmid library again revealed a tandem arrangement of tet(O/32/O) and tet(40), flanked by regions with homology to parts of the VanG operon previously identified in Enterococcus faecalis. At least 10 of the metagenomic inserts that carried tet(O/32/O) also carried tet(40), suggesting that tet(40), although previously undetected, may be an abundant efflux gene.

scholarly journals SmdAB, a Heterodimeric ABC-Type Multidrug Efflux Pump, in Serratia marcescens

2007 ◽  
Vol 190 (2) ◽  
pp. 648-654 ◽  
Author(s):  
Taira Matsuo ◽  
Jing Chen ◽  
Yusuke Minato ◽  
Wakano Ogawa ◽  
Tohru Mizushima ◽  
...  
Keyword(s):  
Serratia Marcescens ◽  
Antimicrobial Agents ◽  
Efflux Pump ◽  
Amino Acid Sequences ◽  
Multidrug Efflux ◽  
Chromosomal Dna ◽  
Atpase Inhibitor ◽  
Multidrug Efflux Pump ◽  
Hoechst 33342 ◽  
E Coli

ABSTRACT We cloned genes, designated smdAB, that encode a multidrug efflux pump from the chromosomal DNA of clinically isolated Serratia marcescens NUSM8906. For cells of the drug-hypersensitive strain Escherichia coli KAM32 harboring a recombinant plasmid carrying smdAB, structurally unrelated antimicrobial agents such as norfloxacin, tetracycline, 4′,6-diamidino-2-phenylindole (DAPI), and Hoechst 33342 showed elevated MICs. The deduced amino acid sequences of both SmdA and SmdB exhibited similarities to the sequences of ATP-binding cassette (ABC)-type multidrug efflux pumps. The efflux of DAPI and Hoechst 33342 from E. coli cells expressing SmdAB was observed, and the efflux activities were inhibited by sodium o-vanadate, which is a well-known ATPase inhibitor. The introduction of smdA or smdB alone into E. coli KAM32 did not elevate the MIC of DAPI; thus, both SmdA and SmdB were required for function. These results indicate that SmdAB is probably a heterodimeric multidrug efflux pump of the ABC family in S. marcescens.

scholarly journals Identification and Sequence of a tet(M) Tetracycline Resistance Determinant Homologue in Clinical Isolates of Escherichia coli

2006 ◽  
Vol 188 (20) ◽  
pp. 7151-7164 ◽  
Author(s):  
C. Hal Jones ◽  
Margareta Tuckman ◽  
Ellen Murphy ◽  
Patricia A. Bradford
Keyword(s):  
Escherichia Coli ◽  
Tetracycline Resistance ◽  
Mobile Element ◽  
Clinical Isolates ◽  
Resistance Determinant ◽  
M Gene ◽  
Insertion Element ◽  
E Coli ◽  
Vibrio Salmonicida ◽  
First Time

ABSTRACT The presence of the tetracycline resistance determinant tet(M) in human clinical isolates of Escherichia coli is described for the first time in this report. The homologue was >99% identical to the tet(M) genes reported to occur in Lactobacillus plantarum, Neisseria meningitidis, and Streptococcus agalactiae, and 3% of the residues in its deduced amino acid sequence diverge from tet(M) of Staphylococcus aureus. Sequence analysis of the regions immediately flanking the gene revealed that sequences upstream of tet(M) in E. coli have homology to Tn916; however, a complete IS26 insertion element was present immediately upstream of the promoter element. Downstream from the termination codon is an insertion sequence that was homologous to the ISVs1 element reported to occur in a plasmid from Vibrio salmonicida that has been associated with another tetracycline resistance determinant, tet(E). Results of mating experiments demonstrated that the E. coli tet(M) gene was on a mobile element so that resistance to tetracycline and minocycline could be transferred to a susceptible strain by conjugation. Expression of the cloned tet(M) gene, under the control of its own promoter, provided tetracycline and minocycline resistance to the E. coli host.

scholarly journals Tet 42, a Novel Tetracycline Resistance Determinant Isolated from Deep Terrestrial Subsurface Bacteria

2008 ◽  
Vol 52 (12) ◽  
pp. 4518-4521 ◽  
Author(s):  
Mindy G. Brown ◽  
Elizabeth H. Mitchell ◽  
David L. Balkwill
Keyword(s):  
Efflux Pump ◽  
Sequence Similarity ◽  
Tetracycline Resistance ◽  
Transmembrane Domains ◽  
Resistance Determinant ◽  
Deep Subsurface ◽  
Terrestrial Subsurface ◽  
Tetr Repressor

ABSTRACT Tet 42, a novel tetracycline resistance determinant from deep subsurface bacteria, was characterized and found to have a 30% sequence similarity to TetA(Z). The protein is a putative efflux pump that shares characteristics with previously characterized pumps, including a divergently transcribed TetR repressor, a conserved GxxSDRxGRR motif, and transmembrane domains.

scholarly journals Integrating Conjugative Elements as Vectors of Antibiotic, Mercury, and Quaternary Ammonium Compound Resistance in Marine Aquaculture Environments

2012 ◽  
Vol 56 (5) ◽  
pp. 2619-2626 ◽  
Author(s):  
Arturo Rodríguez-Blanco ◽  
Manuel L. Lemos ◽  
Carlos R. Osorio
Keyword(s):  
Quaternary Ammonium ◽  
Hot Spot ◽  
Tetracycline Resistance ◽  
Variable Region ◽  
Mercury Resistance ◽  
Ammonium Compound ◽  
Resistance Locus ◽  
Bacterial Strains ◽  
Marine Aquaculture ◽  
Content Type

ABSTRACTThe presence of SXT/R391-related integrating conjugative elements (ICEs) in bacterial strains isolated from fish obtained from marine aquaculture environments in 2001 to 2010 in the northwestern Iberian Peninsula was studied. ICEs were detected in 12 strains taxonomically related toVibrio scophthalmi(3 strains),Vibrio splendidus(5 strains),Vibrio alginolyticus(1 strain),Shewanella haliotis(1 strain), andEnterovibrio nigricans(2 strains), broadening the known host range able to harbor SXT/R391-like ICEs. Variable DNA regions, which confer element-specific properties to ICEs of this family, were characterized. One of the ICEs encoded antibiotic resistance functions in variable region III, consisting of a tetracycline resistance locus. Interestingly, hot spot 4 included genes providing resistance to rifampin (ICEVspPor2 and ICEValPor1) and quaternary ammonium compounds (QACs) (ICEEniSpa1), and variable region IV included a mercury resistance operon (ICEVspSpa1 and ICEEniSpa1). The S exclusion group was more represented than the R exclusion group, accounting for two-thirds of the total ICEs. Mating experiments allowed ICE mobilization toEscherichia colistrains, showing the corresponding transconjugants' rifampin, mercury, and QAC resistance. These results show the first evidence of ICEs providing rifampin and QAC resistances, suggesting that these mobile genetic elements contribute to the dissemination of antimicrobial, heavy metal, and QAC resistance determinants in aquaculture environments.

scholarly journals Burkholderia ubonensis high-level tetracycline resistance is due to efflux pump synergy involving a novel TetA(64) resistance determinant

2020 ◽  
pp. AAC.01767-20
Author(s):  
Nawarat Somprasong ◽  
Carina M. Hall ◽  
Jessica R. Webb ◽  
Jason W. Sahl ◽  
David M. Wagner ◽  
...  
Keyword(s):  
Burkholderia Cepacia ◽  
Efflux Pump ◽  
Tetracycline Resistance ◽  
Major Facilitator Superfamily ◽  
Resistance Determinant ◽  
Chromosome 1 ◽  
Human Pathogens ◽  
Clinically Significant ◽  
Major Facilitator ◽  
High Level

Burkholderia ubonensis, a non-pathogenic soil bacterium belonging to the Burkholderia cepacia complex (Bcc), is highly resistant to some clinically significant antibiotics. The concern is that B. ubonensis may serve as a resistance reservoir for Bcc or B. pseudomallei complex (Bpc) organisms that are opportunistic human pathogens. Using a B. ubonensis strain highly resistant to tetracycline (MIC ≥256 μg/ml), we identified and characterized tetA(64) that encodes a novel tetracycline-specific efflux pump of the major facilitator superfamily. TetA(64) and associated TetR(64) regulator expression is induced by tetracyclines. Although TetA(64) is the primary tetracycline and doxycycline resistance determinant, maximum tetracycline and doxycycline resistance requires synergy between TetA(64) and the non-specific AmrAB-OprA resistance nodulation cell division efflux pump. TetA(64) does not efflux minocycline, tigecycline and eravacycline. Comprehensive screening of genome sequences showed that TetA(64) is unequally distributed in the Bcc and absent from the Bpc. It is present in some major cystic fibrosis pathogens, like B. cenocepacia, but absent from others like B. multivorans. The tetR(64)-tetA(64) genes are located in a region of chromosome 1 that is highly conserved in Burkholderia. Because there is no evidence for transposition, the tetR(64)-tetA(64) genes may have been acquired by homologous recombination after horizontal gene transfer. Although Burkholderia species contain a resident multi-component efflux pump that allows them to respond to tetracyclines up to a certain concentration, the acquisition of the single-component TetA(64) by some species likely provides the synergy that these bacteria need to defend against high tetracycline concentrations in niche environments.

scholarly journals An arsRB resistance operon confers tolerance to arsenite in the environmental isolate Terribacillus sp. AE2B 122

2021 ◽  
Author(s):  
Almudena Escobar-Niño ◽  
Leyre Sánchez-Barrionuevo ◽  
José Miguel Torres-Torres ◽  
Rafael Clemente ◽  
Gabriel Gutiérrez ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Escherichia Coli ◽  
Heterologous Expression ◽  
Efflux Pump ◽  
Arsenic Speciation ◽  
Transcriptional Regulator ◽  
Environmental Strain ◽  
E Coli ◽  
Ars Operon ◽  
Arsenate Reduction

Abstract Terribacillus sp. AE2B 122 is an environmental strain isolated from olive-oil agroindustry wastes. This strain displays resistance to arsenic, one of the most ubiquitous carcinogens found in nature. Terribacillus sp. AE2B 122 possesses an unusual ars operon, consisting of the transcriptional regulator (arsR) and arsenite efflux pump (arsB) but no adjacent arsenate reductase (arsC) locus. Expression of arsR and arsB was induced when Terribacillus was exposed to sub-lethal concentrations of arsenate. Heterologous expression of the arsB homologue in Escherichia coli ∆arsRBC demonstrated that it conferred resistance to arsenite and reduced the accumulation of arsenic inside the cells. Two members of the arsC-like family (Te3384 and Te2854) found in the Terribacillus genome were not induced by arsenic, but their heterologous expression in E. coli ∆arsC and ∆arsRBC increased the accumulation of arsenic in both strains. We found that both Te3384 and Te2854 slightly increased resistance to arsenate in E. coli ∆arsC and ∆arsRBC, possibly by chelation of arsenic or increasing the resistance to oxidative stress. Finally, arsenic speciation assays suggest that Terribacillus is incapable of arsenate reduction, in agreement with the lack of an arsC homologue in the genome.

scholarly journals Identification and Transferability of Tetracycline Resistance in Streptococcus thermophilus during Milk Fermentation, Storage, and Gastrointestinal Transit

Fermentation ◽  
2021 ◽  
Vol 7 (2) ◽  
pp. 65
Author(s):  
Armin Tarrah ◽  
Shadi Pakroo ◽  
Viviana Corich ◽  
Alessio Giacomini
Keyword(s):  
Efflux Pump ◽  
Acquired Resistance ◽  
Gastrointestinal Transit ◽  
Tetracycline Resistance ◽  
Lactobacillus Delbrueckii ◽  
Genomic Islands ◽  
Resistant Bacteria ◽  
Human Pathogens ◽  
Resistance Level ◽  
Milk Fermentation

The existence of antibiotic-resistant bacteria in food products, particularly those carrying acquired resistance genes, has increased concerns about the transmission of these genes from beneficial microbes to human pathogens. In this study, we evaluated the antibiotic resistance-susceptibility patterns of 16 antibiotics in eight S. thermophilus strains, whose genome sequence is available, using phenotypic and genomic approaches. The minimal inhibitory concentration values collected revealed intermediate resistance to aminoglycosides, whereas susceptibility was detected for different classes of β-lactams, quinolones, glycopeptide, macrolides, and sulfonamides in all strains. A high tetracycline resistance level has been detected in strain M17PTZA496, whose genome analysis indicated the presence of the tet(S) gene and the multidrug and toxic compound extrusion (MATE) family efflux pump. Moreover, an in-depth genomic analysis revealed genomic islands and an integrative and mobilizable element (IME) in the proximity of the gene tet(S). However, despite the presence of a prophage, genomic islands, and IME, no horizontal gene transfer was detected to Lactobacillus delbrueckii subsp. lactis DSM 20355 and Lactobacillusrhamnosus GG during 24 h of skim milk fermentation, 2 weeks of refrigerated storage, and 4 h of simulated gastrointestinal transit.

scholarly journals Insight into the AcrAB-TolC Complex Assembly Process Learned from Competition Studies

Antibiotics ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 830
Author(s):  
Prasangi Rajapaksha ◽  
Isoiza Ojo ◽  
Ling Yang ◽  
Ankit Pandeya ◽  
Thilini Abeywansha ◽  
...  
Keyword(s):  
Efflux Pump ◽  
Dominant Negative ◽  
Dominant Negative Effect ◽  
Assembly Process ◽  
Multidrug Efflux ◽  
E Coli ◽  
Multidrug Efflux Pumps ◽  
Negative Effect ◽  
Dynamic Assembly ◽  
Pump Assembly

The RND family efflux pump AcrAB-TolC in E. coli and its homologs in other Gram-negative bacteria are major players in conferring multidrug resistance to the cells. While the structure of the pump complex has been elucidated with ever-increasing resolution through crystallography and Cryo-EM efforts, the dynamic assembly process remains poorly understood. Here, we tested the effect of overexpressing functionally defective pump components in wild type E. coli cells to probe the pump assembly process. Incorporation of a defective component is expected to reduce the efflux efficiency of the complex, leading to the so called “dominant negative” effect. Being one of the most intensively studied bacterial multidrug efflux pumps, many AcrA and AcrB mutations have been reported that disrupt efflux through different mechanisms. We examined five groups of AcrB and AcrA mutants, defective in different aspects of assembly and substrate efflux. We found that none of them demonstrated the expected dominant negative effect, even when expressed at concentrations many folds higher than their genomic counterpart. The assembly of the AcrAB-TolC complex appears to have a proof-read mechanism that effectively eliminated the formation of futile pump complex.

scholarly journals Bioactive constituents with antibacterial, resistance modulation, anti-biofilm formation and efflux pump inhibition properties from Aidia genipiflora stem bark

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Daniel Anokwah ◽  
Evelyn Asante-Kwatia ◽  
Abraham Y. Mensah ◽  
Cynthia Amaning Danquah ◽  
Benjamin K. Harley ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Efflux Pump ◽  
Ethyl Acetate Fraction ◽  
Stem Bark ◽  
Resistance Mechanisms ◽  
Bioactive Constituents ◽  
Biofilm Inhibition ◽  
E Coli ◽  
Efflux Pump Inhibition ◽  
Oleanonic Acid

Abstract Background Antimicrobial resistance is a global health challenge. The involvement of bacterial biofilms and efflux pumps in the development of multidrug resistance (MDR) is well established. Medicinal plants have been proposed as alternatives for combating MDR focusing on their bioactive constituents with resistance modulatory activities. This study was aimed at investigating the stem bark of Aidia genipiflora for bioactive constituents with anti-biofilm, efflux pump inhibition and resistance modulatory activities. Method The crude methanol extract was purified by column chromatography and isolated compounds characterized by mass and nuclear magnetic resonance spectrometry. Antibacterial activity was determined by the High-throughput spot culture growth inhibition and the broth micro-dilution assay. The ethidium bromide accumulation assay was used to determine efflux pump inhibition property. Biofilm inhibition was determined in a microplate crystal violet retention assay. Results Purification of the ethyl acetate fraction led to the isolation of oleanonic acid (1), 4-hydroxy cinnamic acid docosyl ester (2), β-stigmasterol/β-sitosterol (mixture 3a/b) and D-mannitol (4). The minimum inhibitory concentrations (MICs) ranged from 250 to > 500 μg/mL for extracts and fractions and from 15 to 250 μg/mL for compounds. In the presence of sub-inhibitory concentrations of the compounds, the MIC of amoxicillin against E. coli (20 μg/mL) and P. aeruginosa (320 μg/mL) was reduced by 32 and 10 folds respectively. The whole extract demonstrated anti-biofilm formation and efflux pump inhibition in E. coli, S. aureus and P. aeruginosa. The sterol mixture (3a/b) at concentration of 100 μg/mL caused the highest inhibition (73%) of biofilm formation in S. aureus. Oleanonic acid (1) demonstrated remarkable efflux pump inhibition at MIC of 7.8 μg/mL in E. coli better than the standard drugs verapamil and chlorpromazine. Conclusion This study confirms the prospects of A. genipiflora as a source of new antibacterial agents and adjuvants that could interact with some resistance mechanisms in bacteria to enhance the activity of hitherto ineffective antibiotics. “A small portion of the study has been presented in a conference in the form of poster”.

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