Neither reduced uptake nor increased efflux is encoded by tellurite resistance determinants expressed inEscherichia coli

1995 ◽  
Vol 41 (1) ◽  
pp. 92-98 ◽  
Author(s):  
Raymond J. Turner ◽  
Joel H. Weiner ◽  
Diane E. Taylor
Keyword(s):  
Resistance Mechanism ◽  
Metal Resistance ◽  
Resistance Determinant ◽  
Inescherichia Coli ◽  
Efflux Transporter ◽  
Tellurite Resistance ◽  
E Coli ◽  
Resistance Determinants ◽  
Escherichia Coli Cells ◽  
Resistance Plasmids

Rates of uptake of the TeO32−oxyanion were investigated in Escherichia coli cells containing tellurite resistance determinants from both plasmid (RK2Ter, R478, pMER610, MIP233, pHH1508a, pMUR) and chromosomal (tehAB) sources. The uptake was investigated to determine whether or not reduced uptake or increased efflux is involved in the tellurite resistance mechanism. Reduced TeO32−uptake generated by cultures harboring arsABC from the plasmid R773, which has been previously shown to be an oxyanion efflux transporter, was used as the standard. Uptake curves were found to be essentially identical among E. coli cultures harboring the tellurite resistance plasmids RK2Ter, pMER610, pHH1508a, and pMUR and cultures harboring tellurite-sensitive control plasmids. Cultures harboring clones of the tehAB operon from E. coli showed no change in the TeO32−accumulation. Cultures harboring R478 demonstrated reduced uptake. However, a subclone containing only the tellurite resistance determinant displayed no reduced uptake. This suggests that there may be another determinant on R478 other than the primary tellurite resistance determinant that gives rise to TeO32−efflux. These results demonstrate that neither reduced uptake nor increased efflux is responsible for the tellurite resistance in the resistance determinants investigated here.Key words: tellurite resistance, uptake, metal resistance, resistance.

Glutathione is a target in tellurite toxicity and is protected by tellurite resistance determinants inEscherichia coli

2001 ◽  
Vol 47 (1) ◽  
pp. 33-40 ◽  
Author(s):  
Raymond J Turner ◽  
Yair Aharonowitz ◽  
Joel H Weiner ◽  
Diane E Taylor
Keyword(s):  
Reduced Glutathione ◽  
Resistance Mechanism ◽  
Thiol Oxidation ◽  
Wild Type ◽  
Response Curves ◽  
Tellurite Resistance ◽  
Thiol Compounds ◽  
Mechanisms Of Toxicity ◽  
Resistance Determinants ◽  
Glutathione Oxidation

Tellurite (TeO32-) is highly toxic to most microorganisms. The mechanisms of toxicity or resistance are poorly understood. It has been shown that tellurite rapidly depletes the reduced thiol content within wild-type Escherichia coli. We have shown that the presence of plasmid-borne tellurite-resistance determinants protects against general thiol oxidation by tellurite. In the present study we observe that the tellurite-dependent depletion of cellular thiols in mutants of the glutathione and thioredoxin thiol:redox system was less than in wild-type cells. To identify the type of low-molecular-weight thiol compounds affected by tellurite exposure, the thiol-containing molecules were analyzed by reverse phase HPLC as their monobromobimane derivatives. Results indicated that reduced glutathione is a major initial target of tellurite reactivity within the cell. Other thiol species are also targeted by tellurite, including reduced coenzyme A. The presence of the tellurite resistance determinants kilA and ter protect against the loss of reduced glutathione by as much as 60% over a 2 h exposure. This protection of glutathione oxidation is likely key to the resistance mechanism of these determinants. Additionally, the thiol oxidation response curves were compared between selenite and tellurite. The loss of thiol compounds within the cell recovered from selenite but not to tellurite.Key words: tellurite, resistance, thiol oxidation, heavy metal toxicity, selenite, glutathione.

scholarly journals Apramycin resistance plasmids inEscherichia coli: possible transfer toSalmonella typhimuriumin calves

1992 ◽  
Vol 108 (2) ◽  
pp. 271-278 ◽  
Author(s):  
J. E. B. Hunter ◽  
J. C. Shelley ◽  
J. R. Walton ◽  
C. A. Hart ◽  
M. Bennett
Keyword(s):  
Escherichia Coli ◽  
Salmonella Typhimurium ◽  
High Frequency ◽  
Inescherichia Coli ◽  
Gene Encoding ◽  
E Coli ◽  
Resistance Plasmids ◽  
Effect Of Treatment ◽  
K 12

SUMMARYAn outbreak of salmonellosis in calves was monitored for persistence ofSalmonella typhimuriumexcretion in faeces and the effect of treatment with apramycin. Prior to treatment apramycin-resistantEscherichia coliwere present but allS. typhimuriumisolates were sensitive. Following the treatment of six calves with apramycin, apramycin-resistantS. typhimuriumwere isolated from two treated calves and one untreated calf. Plasmid profiles ofE. coliandS. typhimuriumwere compared and plasmids conferring resistance to apramycin and several other antibiotics were transferred by conjugationin vitrofrom calfE. coliandS. typhimuriumisolates toE. coliK-12 and fromE. colitoS. typhimurium. The plasmids conjugated with high frequencyin vitrofromE. colitoS. typhimurium, and hybridized to a DNA probe specific for the gene encoding aminoglycoside acetyltransferase 3-IV (AAC(3)-IV) which confers resistance to apramycin, gentamicin, netilmicin and tobramycin.

scholarly journals Evaluation of the Shift in Antimicrobial Resistance Due to Extended Spectrum Beta-Lactamase and AmpC Producing Enterobacteriaceae in Hampshire England

2018 ◽  
Vol 1 (1) ◽  
Author(s):  
Sarah Fouch
Keyword(s):  
Antimicrobial Resistance ◽  
Resistance Mechanism ◽  
Community Setting ◽  
Resistance Determinant ◽  
Antibiotic Prescribing ◽  
Esbl Production ◽  
Patient Age ◽  
Extended Spectrum Beta Lactamase ◽  
Resistance Determinants ◽  
Patient Âgé

The aim of this study was to identify shifts in antimicrobial resistance in the Hampshire region including correlations between patient demographic and antibiotic prescribing to inform safe and effective antimicrobial stewardship. 475 ESBL and AmpC producing bacteria, from various infection sites, were obtained from four hospital laboratories in Hampshire, UK, during 2010 and 2012. All isolates were identified to species level. ESBL production and antimicrobial susceptibility testing was performed using disc diffusion methodology. Multiplex PCR and gel electrophoresis was used to detect the BlaCTX, BlaTEM and BlaSHV resistance determinants. Corresponding patient data included patient age, gender, location, clinical details and previous antibiotic therapy. Patient information revealed mean ages of 60 and 62 for the 2010 and 2012 cohorts respectively, with ages ranging from 3 months to 96. ESBL production was the most prevalent resistance mechanism (65% in 2010, 79% in 2012), produced mostly by E.coli (85% in 2010, 84% in 2012). While 9 of the 13 antibiotics demonstrated increased resistance, 4 demonstrated a decrease. CTX was the most prevalent resistance determinant (38% in 2010 and 27% in 2012), followed by dual expression of CTX & TEM, TEM, TEM & SHV, SHV, SHV & CTX and all three resistance determinants. A significant correlation between patient age and joint expression of TEM & SHV was observed in 2010, in 2012, patient age significantly correlated with joint CTX and SHV expression. Significant differences could also be determined between resistance determinant type and antimicrobial resistance. This study shows that the incidence of ESBL and AmpC infections and resistance to commonly used antibiotics within Hampshire is increasing both within the hospital and community setting. This emphasises the need for judicious antibiotic prescribing to safeguard this valuable medical commodity.

scholarly journals Replication of Staphylococcal Multiresistance Plasmids

2000 ◽  
Vol 182 (8) ◽  
pp. 2170-2178 ◽  
Author(s):  
Neville Firth ◽  
Sumalee Apisiridej ◽  
Tracey Berg ◽  
Brendon A. O'Rourke ◽  
Steve Curnock ◽  
...  
Keyword(s):  
Sequence Similarity ◽  
Heavy Metal Resistance ◽  
Metal Resistance ◽  
Replication Initiation ◽  
Conjugative Plasmid ◽  
Rolling Circle ◽  
Segregational Stability ◽  
Structural And Functional Properties ◽  
Resistance Plasmids ◽  
Genes Encoding

ABSTRACT Based on structural and functional properties, three groups of large staphylococcal multiresistance plasmids have been recognized, viz., the pSK1 family, pSK41-like conjugative plasmids, and β-lactamase–heavy-metal resistance plasmids. Here we describe an analysis of the replication functions of a representative of each of these plasmid groups. The replication initiation genes from theStaphylococcus aureus plasmids pSK1, pSK41, and pI9789::Tn552 were found to be related to each other and to the Staphylococcus xylosus plasmid pSX267 and are also related to rep genes of several plasmids from other gram-positive genera. Nucleotide sequence similarity between pSK1 and pI9789::Tn552 extended beyond theirrep genes, encompassing upstream divergently transcribed genes, orf245 and orf256, respectively. Our analyses revealed that genes encoding proteins related to the deducedorf245 product are variously represented, in several types of organization, on plasmids possessing six seemingly evolutionarily distinct types of replication initiation genes and including both theta-mode and rolling-circle replicons. Construction of minireplicons and subsequent functional analysis demonstrated that orf245is required for the segregational stability of the pSK1 replicon. In contrast, no gene equivalent to orf245 is evident on the conjugative plasmid pSK41, and a minireplicon encoding only the pSK41 rep gene was found to exhibit a segregational stability approaching that of the parent plasmid. Significantly, the results described establish that many of the large multiresistance plasmids that have been identified in clinical staphylococci, which were formerly presumed to be unrelated, actually utilize an evolutionarily related theta-mode replication system.

scholarly journals Impact of Feed Supplementation with Antimicrobial Agents on Growth Performance of Broiler Chickens, Clostridium perfringens and Enterococcus Counts, and Antibiotic Resistance Phenotypes and Distribution of Antimicrobial Resistance Determinants in Escherichia coli Isolates

2007 ◽  
Vol 73 (20) ◽  
pp. 6566-6576 ◽  
Author(s):  
Moussa S. Diarra ◽  
Fred G. Silversides ◽  
Fatoumata Diarrassouba ◽  
Jane Pritchard ◽  
Luke Masson ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Antimicrobial Resistance ◽  
Broiler Chickens ◽  
Antimicrobial Agents ◽  
Growth Promoters ◽  
Feed Supplementation ◽  
E Coli ◽  
Resistance Determinants ◽  
Class 1

ABSTRACT The effects of feed supplementation with the approved antimicrobial agents bambermycin, penicillin, salinomycin, and bacitracin or a combination of salinomycin plus bacitracin were evaluated for the incidence and distribution of antibiotic resistance in 197 commensal Escherichia coli isolates from broiler chickens over 35 days. All isolates showed some degree of multiple antibiotic resistance. Resistance to tetracycline (68.5%), amoxicillin (61.4%), ceftiofur (51.3%), spectinomycin (47.2%), and sulfonamides (42%) was most frequent. The levels of resistance to streptomycin, chloramphenicol, and gentamicin were 33.5, 35.5, and 25.3%, respectively. The overall resistance levels decreased from day 7 to day 35 (P < 0.001). Comparing treatments, the levels of resistance to ceftiofur, spectinomycin, and gentamicin (except for resistance to bacitracin treatment) were significantly higher in isolates from chickens receiving feed supplemented with salinomycin than from the other feeds (P < 0.001). Using a DNA microarray analysis capable of detecting commonly found antimicrobial resistance genes, we characterized 104 tetracycline-resistant E. coli isolates from 7- to 28-day-old chickens fed different growth promoters. Results showed a decrease in the incidence of isolates harboring tet(B), bla TEM, sulI, and aadA and class 1 integron from days 7 to 35 (P < 0.01). Of the 84 tetracycline-ceftiofur-resistant E. coli isolates, 76 (90.5%) were positive for bla CMY-2. The proportions of isolates positive for sulI, aadA, and integron class 1 were significantly higher in salinomycin-treated chickens than in the control or other treatment groups (P < 0.05). These data demonstrate that multiantibiotic-resistant E. coli isolates can be found in broiler chickens regardless of the antimicrobial growth promoters used. However, the phenotype and the distribution of resistance determinants in E. coli can be modulated by feed supplementation with some of the antimicrobial agents used in broiler chicken production.

scholarly journals Genetic but No Phenotypic Associations between Biocide Tolerance and Antibiotic Resistance in Escherichia coli from German Broiler Fattening Farms

2021 ◽  
Vol 9 (3) ◽  
pp. 651
Author(s):  
Alice Roedel ◽  
Szilvia Vincze ◽  
Michaela Projahn ◽  
Uwe Roesler ◽  
Caroline Robé ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Acquired Resistance ◽  
Antibiotic Resistance Genes ◽  
Animal Husbandry ◽  
Metal Resistance ◽  
Resistant Bacteria ◽  
Virulence Determinants ◽  
Phylogenetic Distribution ◽  
Resistance Determinants

Biocides are frequently applied as disinfectants in animal husbandry to prevent the transmission of drug-resistant bacteria and to control zoonotic diseases. Concerns have been raised, that their use may contribute to the selection and persistence of antimicrobial-resistant bacteria. Especially, extended-spectrum β-lactamase- and AmpC β-lactamase-producing Escherichia coli have become a global health threat. In our study, 29 ESBL-/AmpC-producing and 64 NON-ESBL-/AmpC-producing E.coli isolates from three German broiler fattening farms collected in 2016 following regular cleaning and disinfection were phylogenetically characterized by whole genome sequencing, analyzed for phylogenetic distribution of virulence-associated genes, and screened for determinants of and associations between biocide tolerance and antibiotic resistance. Of the 30 known and two unknown sequence types detected, ST117 and ST297 were the most common genotypes. These STs are recognized worldwide as pandemic lineages causing disease in humans and poultry. Virulence determinants associated with extraintestinal pathogenic E.coli showed variable phylogenetic distribution patterns. Isolates with reduced biocide susceptibility were rarely found on the tested farms. Nine isolates displayed elevated MICs and/or MBCs of formaldehyde, chlorocresol, peroxyacetic acid, or benzalkonium chloride. Antibiotic resistance to ampicillin, trimethoprim, and sulfamethoxazole was most prevalent. The majority of ESBL-/AmpC-producing isolates carried blaCTX-M (55%) or blaCMY-2 (24%) genes. Phenotypic biocide tolerance and antibiotic resistance were not interlinked. However, biocide and metal resistance determinants were found on mobile genetic elements together with antibiotic resistance genes raising concerns that biocides used in the food industry may lead to selection pressure for strains carrying acquired resistance determinants to different antimicrobials.

scholarly journals In Vitro and In Vivo Antibacterial Activities of a Novel Glycylcycline, the 9-t-Butylglycylamido Derivative of Minocycline (GAR-936)

1999 ◽  
Vol 43 (4) ◽  
pp. 738-744 ◽  
Author(s):  
P. J. Petersen ◽  
N. V. Jacobus ◽  
W. J. Weiss ◽  
P. E. Sum ◽  
R. T. Testa
Keyword(s):  
Anaerobic Bacteria ◽  
Tetracycline Resistance ◽  
Protective Effects ◽  
Gram Negative ◽  
E Coli ◽  
Resistance Determinants ◽  
Aerobic And Anaerobic ◽  
Ribosomal Protection

ABSTRACT The 9-t-butylglycylamido derivative of minocycline (TBG-MINO) is a recently synthesized member of a novel group of antibiotics, the glycylcyclines. This new derivative, like the first glycylcyclines, theN,N-dimethylglycylamido derivative of minocycline and 6-demethyl-6-deoxytetracycline, possesses activity against bacterial isolates containing the two major determinants responsible for tetracycline resistance: ribosomal protection and active efflux. The in vitro activities of TBG-MINO and the comparative agents were evaluated against strains with characterized tetracycline resistance as well as a spectrum of recent clinical aerobic and anaerobic gram-positive and gram-negative bacteria. TBG-MINO, with an MIC range of 0.25 to 0.5 μg/ml, showed good activity against strains expressing tet(M) (ribosomal protection), tet(A), tet(B),tet(C), tet(D), and tet(K) (efflux resistance determinants). TBG-MINO exhibited similar activity against methicillin-resistant Staphylococcus aureus (MRSA), penicillin-resistant streptococci, and vancomycin-resistant enterococci (MICs at which 90% of strains are inhibited, ≤0.5 μg/ml). TBG-MINO exhibited activity against a wide diversity of gram-negative aerobic and anaerobic bacteria, most of which were less susceptible to tetracycline and minocycline. The in vivo protective effects of TBG-MINO were examined against acute lethal infections in mice caused by Escherichia coli, S. aureus, andStreptococcus pneumoniae isolates. TBG-MINO, administered intravenously, demonstrated efficacy against infections caused byS. aureus including MRSA strains and strains containingtet(K) or tet(M) resistance determinants (median effective doses [ED50s], 0.79 to 2.3 mg/kg of body weight). TBG-MINO demonstrated efficacy against infections caused by tetracycline-sensitive E. coli strains as well asE. coli strains containing either tet(M) or the efflux determinant tet(A), tet(B), ortet(C) (ED50s, 1.5 to 3.5 mg/kg). Overall, TBG-MINO shows antibacterial activity against a wide spectrum of gram-positive and gram-negative aerobic and anaerobic bacteria including strains resistant to other chemotherapeutic agents. The in vivo protective effects, especially against infections caused by resistant bacteria, corresponded with the in vitro activity of TBG-MINO.

scholarly journals The characterization of ESBL genes in Escherichia coli and Klebsiella pneumoniae causing nosocomial infections in Vietnam

2013 ◽  
Vol 7 (12) ◽  
pp. 922-928 ◽  
Author(s):  
Nguyen Hoang Thu Trang ◽  
Tran Vu Thieu Nga ◽  
James I Campbell ◽  
Nguyen Trong Hiep ◽  
Jeremy Farrar ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Klebsiella Pneumoniae ◽  
Nosocomial Infections ◽  
Enteric Bacteria ◽  
E Coli ◽  
Plasmid Analysis ◽  
Resistance Plasmids ◽  
Genes Encoding ◽  
Third Generation Cephalosporins

Background: Extended-spectrum β-lactamases (ESBLs) are enzymes capable of hydrolyzing oxyimino-β-lactams and inducing resistance to third generation cephalosporins. The genes encoding ESBLs are widespread and generally located on highly transmissible resistance plasmids. We aimed to investigate the complement of ESBL genes in E. coli and Klebsiella pneumoniae causing nosocomial infections in hospitals in Ho Chi Minh City, Vietnam. Methodology: Thirty-two non-duplicate isolates of E. coli and Klebsiella pneumoniae causing nosocomial infections, isolated between March and June 2010, were subjected to antimicrobial susceptibility testing. All isolates were PCR-amplified to detect the blaSHV, blaTEM and blaCTX-M ESBL genes and subjected to plasmid analysis. Results: We found that co-resistance to multiple antimicrobials was highly prevalent, and we report the predominance of the blaCTX-M-15 and blaCTX-M-27 genes, located on highly transmissible plasmids ranging from 50 to 170 kb in size. Conclusions: Our study represents a snap shot of ESBL-producing enteric bacteria causing nosocomial infections in this setting. We suggest that antimicrobial resistance in nosocomial E. coli and Klebsiella pneumoniae is rampant in Vietnam and ESBL organisms are widespread. In view of these data and the dramatic levels of antimicrobial resistance reported in Vietnam we advocate an urgent review of antimicrobial use in the Vietnamese healthcare system.

scholarly journals Induction and control of the autolytic system of Escherichia coli

1982 ◽  
Vol 152 (1) ◽  
pp. 26-34
Author(s):  
M Leduc ◽  
R Kasra ◽  
J van Heijenoort
Keyword(s):  
Escherichia Coli ◽  
Fatty Acid ◽  
Acid Synthesis ◽  
Induction Method ◽  
E Coli ◽  
Escherichia Coli Cells ◽  
Carbonyl Cyanide ◽  
And Control ◽  
First Time ◽  
Induced Autolysis

Various methods of inducing autolysis of Escherichia coli cells were investigated, some being described here for the first time. For the autolysis of growing cells only induction methods interfering with the biosynthesis of peptidoglycan were taken into consideration, whereas with harvested cells autolysis was induced by rapid osmotic or EDTA shock treatments. The highest rates of autolysis were observed after induction by moenomycin, EDTA, or cephaloridine. The different autolyses examined shared certain common properties. In particular, regardless of the induction method used, more or less extensive peptidoglycan degradation was observed, and 10(-2) M Mg2+ efficiently inhibited the autolytic process. However, for other properties a distinction was made between methods used for growing cells and those used for harvested cells. Autolysis of growing cells required RNA, protein, and fatty acid synthesis. No such requirements were observed with shock-induced autolysis performed with harvested cells. Thus, the effects of Mg2+, rifampicin, chloramphenicol, and cerulenin clearly suggest that distinct factors are involved in the control of the autolytic system of E. Coli. Uncoupling agents such as sodium azide, 2,4-dinitrophenol, and carbonyl-cyanide-m-chlorophenyl hydrazone used at their usual inhibiting concentration had no effect on the cephaloridine or shock-induced autolysis.

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