Resistance In Serratia Marcescens Research Articles

Comprehensive analysis of resistance-nodulation-cell division superfamily (RND) efflux pumps from Serratia marcescens, Db10

We investigated the role of the resistance-nodulation-cell division superfamily (RND) efflux system on intrinsic multidrug resistance in Serratia marcescens. We identified eight putative RND efflux system genes in the S. marcescens Db10 genome that included the previously characterized systems, sdeXY, sdeAB, and sdeCDE. Six out of the eight genes conferred multidrug resistance on KAM32, a drug hypersensitive strain of Escherichia coli. Five out of the eight genes conferred resistance to benzalkonium, suggesting the importance of RND efflux systems in biocide resistance in S. marcescens. The energy-dependent efflux activities of five of the pumps were examined using a rhodamine 6 G efflux assay. When expressed in the tolC-deficient strain of E. coli, KAM43, none of the genes conferred resistance on E. coli. When hasF, encoding the S. marcescens TolC ortholog, was expressed in KAM43, all of the genes conferred resistance on E. coli, suggesting that HasF is a major outer membrane protein that is used by all RND efflux systems in this organism. We constructed a sdeXY deletion mutant from a derivative strain of the clinically isolated multidrug-resistant S. marcescens strain and found that the sdeXY deletion mutant was sensitive to a broad spectrum of antimicrobial agents.

The lack of OmpF, but not OmpC, contributes to increased antibiotic resistance in Serratia marcescens.

The environmental organism Serratia marcescens is one of the primary causes of numerous nosocomial outbreaks and opportunistic infections. Multi-drug resistance is now a common feature among S. marcescens clinical isolates, complicating the efficacy of treatment. Recent reports have attributed antibiotic resistance to altered porin expression as well as perturbation of the intrinsic AmpC beta-lactamase production pathway. In this study, we aimed to genetically correlate the absence of OmpF and OmpC classical porins with increased antibiotic resistance. In generating isogenic porin mutant strains, we avoided incorporating additional resistance through the use of antibiotic cassettes in gene replacement and adopted an alternative strategy in creating clean unmarked mutant strains. We found that lack of OmpF, but not OmpC, significantly increased antibiotic MIC values to the beta-lactam drugs such as ampicillin and cefoxitin as well as to nitrofurantoin. Furthermore, we found that cefoxitin did not induce intrinsic AmpC beta-lactamase production, indicating that the increased MIC values were a result of reduced permeability of cefoxitin due to the lack of OmpF. Genetic deletion of both ompF and ompC did not compromise the integrity of the bacterial cell envelope in optimal growth conditions, suggesting that other outer-membrane porins may function in a compensatory role to facilitate nutrient uptake and cell envelope integrity. Taken together, to our knowledge this is the first study that genetically correlates increased antibiotic resistance with altered porin expression in S. marcescens.

Mechanisms of antimicrobial resistance in Serratia marcescens

Serratia marcescens, one of the Enterobacteriaceae, was considered originally to be non-pathogenic. But in recent decades, nosocomial infections caused by this organism are often hard to treat because of both the intrinsic resistance of this species and its abilities to acquire further resistance to multiple groups of antimicrobial agents, including β-lactams, aminoglycosides and fluoroquinolones. S. marcescens have their own ways to become resistant in common with other Enterobacteriaceae: production of β-lactamasesincluding extended-spectrum β-lactamases, AmpC-type cephalosporinase and carbapenemases; diminished outer membrane permeability; modification of the target site-PBPs; overexpression of active efflux systems; synthesis of aminoglycoside-modifying enzymes; and structural alterations of the GyrA protein. This review will show a brief history, the virulence factors and the known resistance mechanisms to the most frequently administrated antimicrobial agents: β-lactams, aminoglycosides and fluoroquinolones in S. marcescens. Key words: Serratia marcescens, virulence factors, antimicrobial resistance.

Tigecycline resistance in Serratia marcescens associated with up-regulation of the SdeXY-HasF efflux system also active against ciprofloxacin and cefpirome

Efflux by RND-type transporters is known to confer resistance to tigecycline in several Enterobacteriaceae species and we examined the potential of this mechanism in Serratia marcescens using a clinical isolate and laboratory-selected mutants. Expression of RND-type efflux pump genes was analysed by real-time RT-PCR. Laboratory mutants were selected by exposure to either tigecycline or tetracycline in vitro. Efflux pump genes were inactivated by suicide plasmids containing the R6K gamma origin of replication. Higher tigecycline MICs correlated with elevated expression of the RND-type efflux pump genes sdeXY. Inactivation of sdeY or the outer membrane component gene hasF reduced MICs of tigecycline, tetracycline, ciprofloxacin and cefpirome to below those for strain NCTC 10211. A tetracycline-selected laboratory mutant also showed increases in sdeXY expression and tigecycline MIC. Up-regulation of endogenous SdeXY-HasF-mediated efflux is associated with tigecycline resistance in S. marcescens along with MIC rises for tetracycline, ciprofloxacin and cefpirome. Inactivation of this efflux system reduced MICs of those compounds to below those for strain NCTC 10211.

Mutational Upregulation of a Resistance-Nodulation-Cell Division-Type Multidrug Efflux Pump, SdeAB, upon Exposure to a Biocide, Cetylpyridinium Chloride, and Antibiotic Resistance in Serratia marcescens

Serratia marcescens is an important opportunistic pathogen in hospitals, where quaternary ammonium compounds are often used for disinfection. The aim of this study is to elucidate the effect of a biocide on the emergence of biocide- and antibiotic-resistant mutants and to characterize the molecular mechanism of biocide resistance in Serratia marcescens. A quaternary ammonium compound-resistant strain, CRes01, was selected by exposing a wild-type strain of S. marcescens to cetylpyridinium chloride. The CRes01 cells exhibited 2- to 16-fold more resistance than the wild-type cells to biocides and antibiotics, including cetylpyridinium chloride, benzalkonium chloride, chlorhexidine gluconate, fluoroquinolones, tetracycline, and chloramphenicol, and showed increased susceptibilities to beta-lactam antibiotics and N-dodecylpyridinium iodide. Mutant cells accumulated lower levels of norfloxacin than the parent cells in an energized state but not in a de-energized state, suggesting that the strain produced a multidrug efflux pump(s). To verify this assumption, we knocked out a putative efflux pump gene, sdeAB, in CRes01 and found that the knockout restored susceptibility to most quaternary ammonium compounds and antibiotics, to which the CRes01 strain showed resistance. On the basis of these and other results, we concluded that S. marcescens gains resistance to both biocides and antibiotics by expressing the SdeAB efflux pump upon exposure to cetylpyridinium chloride.

Institutional spread of clonally related Serratia marcescens isolates with a novel AmpC cephalosporinase (S4): a 4-year experience in Taiwan

Resistance of Serratia marcescens, a nosocomial pathogen of Enterobacteriaceae, to the extended-spectrum β-lactams is usually mediated by an overproduced AmpC cephalosporinase. We aimed to characterize the molecular epidemiology and AmpC of S. marcescens isolates recovered from 1 medical center in southern Taiwan. AmpC-encoding genes for SRT families were investigated by polymerase chain reaction and DNA sequencing. From August 1999 through July 2003, 69 nonrepetitive bloodstream isolates were enrolled. Excluding 11 isolates, which also produced an extended-spectrum β-lactamase, 58 isolates carried an AmpC-encoding gene, including a novel S4 gene with 98% identity to SRT-1 gene ( n = 50), SRT-2 gene ( n = 3), SST-1 gene ( n = 1), and others ( n = 4). Isolates with S4 exhibited a phenotype of resistance to cefotaxime (CTX) but not ceftazidime. Genotype analysis by pulsed-field gel electrophoresis revealed that 45 (90%) of the isolates carrying S4 gene belonged to 2 major epidemic clones, including types A ( n = 28) and B ( n = 17). In conclusion, the AmpC-like S4 β-lactamase may confer CTX resistance of the S. marcescens population. Strains carrying the S4 gene with prolonged dissemination were closely related.

Characterization of imipenem-resistant Serratia marcescens producing IMP-type and TEM-type β-lactamases encoded on a single plasmid

To evaluate the roles of blaIMP and blaTEM genes in the resistance of Serratia marcescens against beta-lactams and to find the spreading ways of these genes, 19 clinical isolates of imipenem-resistant Serratia marcescens were analyzed. Six strains bore blaIMP and blaTEM genes on a single plasmid, as confirmed by transferring resistance determinants via conjugation and transformation, and by detecting bla genes with PCR analysis. The six strains showed two different genomic patterns on pulsed-field gel electrophoresis. All the transconjugants and transformants gained high-level resistance to ampicillin, cephalexin, cefoxitin and cefotaxime, and showed a reduced susceptibility to imipenem, but maintained full susceptibility to aztreonam. In addition, the expressions of blaIMP and blaTEM genes were constitutive, either in Serratia marcescens clinical isolates or in their transconjugants and transformants. These findings may explain the rapid spread of the above resistance determinants among Enterobacteriaceae via transmissible plasmids in the clinical setting.

Multidrug Resistance in Serratia marcescens and Cloning of Genes Responsible for the Resistance

Six clinically isolated strains of Serratia marcescens were tested for their drug resistance. All showed fairly high resistance to many antimicrobial agents tested including norfloxacin, streptomycin, ampicillin, erythromycin, tetracycline, chloramphenicol, and antimicrobial dyes. Using the drug-hypersensitive strain of Escherichia coli KAM32 as the host, we cloned the genes responsible for multidrug resistance from chromosomal DNA of one of the strains of S. marcescens, NUSM8906. We obtained 28 hybrid plasmids that made host cells resistant to several antimicrobial agents. Many of the transformants harboring each of the plasmids showed multidrug resistance, and some showed resistance to specific drugs. The hybrid plasmids were classified into several groups based on their drug specificity. It appears that each class of plasmid carries different types of drug resistance genes. Analysis of such genes will reveal the multiple mechanisms involved in multidrug resistance in S. marcescens.

Resistance of Serratia marcescens to polymyxin B: a comparative investigation of two S-form lipopolysaccharides obtained from a sensitive and a resistant variant of strain 111

Lipopolysaccharide (LPS) of the polymyxin B (PmB)-sensitive variant of Serratia marcescens smooth-type strain 111 (senLPS) binds significantly more PmB than that of the resistant variant (resLPS), following mild acid hydrolysis this difference is abolished. The main compositional differences between these strains are: (i) the amount of 4-amino-4-deoxy-arabinose (Ara4N); and (ii) the substitution patterns of the phosphate groups. ResLPS contains three times more Ara4N than senLPS, mainly phosphodiesters, whereas senLPS contains mainly (di)-phosphomonoesters. Thus, the senLPS, and the bacterial surface, contain a higher negative charge density. Mild acid hydrolysis decreases the PmB-binding capacity of senLPS and increases that of resLPS. The resulting change in LPS composition was a reduction in the amount of phosphate (about 40%) and a loss of nearly all the Ara4N residues, from both LPS. Mild alkaline hydrolysis removes Ara4N from both LPS, and the hydrolysate contains Ara4N-phosphate, but no Ara4N-(1 →8)-Kdo was detected. The results suggest that the 4'-phosphate of lipid A mediates the binding of PmB, whereas its substitution by Ara4N prevents it.

Spontaneous quinolone resistance in Serratia marcescens due to a mutation in gyrA.

Spontaneous quinolone-resistant mutants of MP050, a quinolone-susceptible clinical strain of Serratia marcescens, were isolated on nutrient agar containing 0.5 microgram of ciprofloxacin per ml. One mutant, designated MP051, was selected for further study. Quinolone MICs for MP051 were 4- to 16-fold higher than those for MP050; nonquinolone MICs were unchanged. The DNA gyrase isolated from MP051 was 24-fold less sensitive to inhibition of supercoiling by ciprofloxacin than the DNA gyrase isolated from MP050 was. Inhibition studies on reconstituted combinations of heterologous gyrase subunits showed that the decreased inhibition was dependent on the A subunit of DNA gyrase from MP051. Further evidence that this decreased inhibition was due to a gyrA mutation was provided by analysis of Escherichia coli gyrA gene expression in S. marcescens heterodiploids containing pNJR3-2, a broad-host-range gyrA gene probe. Quinolone susceptibilities of MP051 heterodiploids containing the wild-type E. coli gyrA gene decreased to those of MP050, while quinolone susceptibilities of MP050 containing the same plasmid were unchanged. These results indicate that spontaneous quinolone resistance in MP051 was due to a mutation in gyrA.

Plasmid-mediated formaldehyde resistance in Serratia marcescens and Escherichia coli: Alterations in the cell surface

The plasmid-mediated formaldehyde resistance of Serratia marcescens and Escherichia coli was examined. For that purpose the outer membranes of isogenic strains (with and without resistance plasmid) were compared. No quantitative or immunological differences in lipopolysaccharide of resistant and sensitive strains were noted. By contrast analysis of outer membrane proteins revealed that the sensitive variants had a higher protein content than the resistant strains. When outer membrane proteins were separated by SDS-PAGE the number of bands seemed identical for sensitive and resistant strains but the intensity of some of the bands was greater for the sensitive isolates. In addition, the surface hydrophobicity was greater for the resistance than for the sensitive strains. These findings suggest that the formaldehyde resistance plasmid of Serratia marcescens confer changes in cell surface proteins and surface hydrophobicity.

Current mechanisms of resistance to antimicrobial agents in microorganisms causing infection in the patient at risk for infection

The mechanisms of resistance encountered in bacteria causing infection in the patient at risk for infection are diverse. Most resistance currently seen is the result of plasmid transfer rather than mutational events. However, extensive use of antimicrobial agents in the hospital has caused the selection of organisms resistant to many agents by virtue of chromosomally mediated mechanisms. Staphylococcus aureus resistant to beta-lactams due to altered penicillin-binding proteins has become a problem in certain patients such as narcotic addicts and chronic care facility patients exposed to many beta-lactam antibiotics. S. epidermidis has also proved to be a problem in patients with indwelling foreign devices, and altered penicillin-binding proteins also make these organisms resistant to available penicillins and cephalosporins. Streptococcus fecalis has become increasingly resistant to aminoglycosides, erythromycin, and tetracyclines due to plasmid-mediated enzymes. Hemophilus influenzae resistant to both penicillins and chloramphenicol by virtue of beta-lactamases and chloramphenicol transacetylase has been encountered. Beta-lactamase-mediated resistance of Enterobacteriaceae, Escherichia coli, and Klebsiella pneumoniae to betalactam antibiotics has increased, and resistance of Serratia marcescens and Pseudomonas aeruginosa to aminoglycosides and penicillins is a widespread phenomenon. Mechanisms to reduce resistance will include not only careful attention to hygienic practices but also more appropriate use of antibiotics selecting the proper agent depending on the type of patient and environment in which the infection develops.

Antibiotic resistance in Serratia marcescens

Antibiotic resistance in Serratia marcescens