Outbreaks of Imipenem Resistant Acinetobacter Baumannii Producing OXA-23 ��-Lactamase in a Tertiary Care Hospital in Korea

Purpose

Since November 2006, imipenem-resistant Acinetobacter baumannii isolates have increased in Kyung Hee University Hospital in Seoul, Korea. The purpose of this study was to determine the genetic basis and molecular epidemiology of outbreak isolates.

Materials and Methods

Forty-nine non-repetitive isolates of the 734 IRAB strains were investigated in order to determine their characteristics. The modified Hodge and the ethylenediaminetetraacetic acid (EDTA)-disk synergy test were performed for the screening of carbapenemase and metallo-β-lactamase production. Multiplex polymerase chain reaction (PCR) assays were performed for the detection of genes encoding for OXA-23-like, OXA-24-like, OXA-58-like and OXA-51-like carbapenemase. Pulsed-field gel electrophoresis (PFGE) was performed for strain identification.

Results

All isolates showed 100% resistance to ciprofloxacin and gentamicin, 97.9% resistance to cefepime, piperacillin/tazobactam, aztreonam, ceftazidime and piperacillin, 93.9% resistance to tobramycin and 57.1% resistance to amikacin. All of the 49 isolates (100%) showed positive results in the modified Hodge test and negative results in the EDTA-disk synergy test. They all (100%) possessed the encoding gene for an intrinsic OXA-51-like carbapenemase and an acquired OXA-23-like carbapenemase in the multiplex PCR assay. PFGE patterns revealed that all isolates were clonally related from A1 to A14.

Conclusion

It is concluded that all of the 49 IRAB isolates acquired resistance to imipenem by producing OXA-23 carbapenemase and they might have originated from a common source.     

In Vivo Selection of Pan-Drug Resistant Acinetobacter baumannii during Antibiotic Treatment

Purpose

Colistin resistance in Acinetobacter baumannii (A. baumannii) is mediated by a complete loss of lipopolysaccharide production via mutations in lpxA, lpxC, and lpxD gene or lipid A modifications via mutations in the pmrA and pmrB genes. However, the exact mechanism of therapy-induced colistin resistance in A. baumannii is not well understood.

Materials and Methods

We investigated the genotypic and phenotypic changes that underlie pan-drug resistance mechanisms by determining differences between the alterations in extensively drug-resistant (XDR) A. baumannii (AB001 and AB002) isolates and a pan-drug resistant (PDR) counterpart (AB003) recovered from one patient before and after antibiotic treatment, respectively.

Results

All three clinical isolates shared an identical sequence type (ST138), belonging to the global epidemic clone, clonal complex 92, and all produced OXA-23 carbapenemase. The PDR AB003 showed two genetic differences, acquisition of armA gene and an amino acid substitution (Glu229Asp) in pmrB gene, relative to XDR isolates. No mutations were detected in the pmrA, pmrC, lpxA, lpxC, or lpxD genes in all three isolates. In matrix-assisted laser desorption ionization-time of flight analysis, the three isolates commonly showed two major peaks at 1728 m/z and 1912 m/z, but peaks at 2034 m/z, 2157 m/z, 2261 m/z, and 2384 m/z were detected only in the PDR A. baumannii AB003 isolate.

Conclusion

Our results show that changes in lipid A structure via a mutation in the pmrB gene and acquisition of armA gene might confer resistance to colistin and aminoglycosides to XDR A. baumannii strains, resulting in appearance of a PDR A. baumannii strain of ST138.     

Clonal spread of carbapenem-resistant Acinetobacter baumannii across a community hospital and its affiliated long-term care facilities: A cross sectional study

Background

The global spread of carbapenem-resistant Acinetobacter baumannii (CRAB) is now a public health problem. In Taiwan, the relationship of the CRAB circulation between long-term care facilities (LTCFs) and acute care hospitals remains unclear. Here, we use molecular epidemiologic methods to describe the transmission of CRAB isolates between a community hospital and its affiliated LTCFs.

Spread of carbapenem-resistant international clones of <Emphasis Type="Italic">Acinetobacter baumannii</Emphasis> in Turkey and Azerbaijan: a collaborative study

Epidemic clones of Acinetobacter baumannii, described as European clones I, II, and III, are associated with hospital epidemics throughout the world. We aimed to determine the molecular characteristics and genetic diversity between European clones I, II, and III from Turkey and Azerbaijan. In this study, a total of 112 bloodstream isolates of carbapenem-resistant Acinetobacter spp. were collected from 11 hospitals across Turkey and Azerbaijan. The identification of Acinetobacter spp. using conventional and sensitivity tests was performed by standard criteria. Multiplex polymerase chain reaction (PCR) was used to detect OXA carbapenemase-encoding genes (bla _OXA-23-like, bla _OXA-24-like, bla _OXA-51-like, and bla _OXA-58-like). Pulsed-field gel electrophoresis (PFGE) typing was used to investigate genetic diversity. The bla _OXA-51-like gene was present in all 112 isolates, 75 (67 %) carried bla _OXA-23-like, 7 (6.2 %) carried bla _OXA-58-like genes, and 5 (4.5 %) carried bla _OXA-24-like genes. With a 90 % similarity cut-off value, 15 clones and eight unique isolates were identified. The largest clone was cluster D, with six subtypes. Isolates from clusters D and I were widely spread in seven different geographical regions throughout Turkey. However, F cluster was found in the northern and eastern regions of Turkey. EU clone I was grouped within J cluster with three isolates found in Antalya, Istanbul, and Erzurum. EU clone II was grouped in the U cluster with 15 isolates and found in Kayseri and Diyarbak?r. The bla _OXA-24-like gene in carbapenemases was identified rarely in Turkey and has been reported for the first time from Azerbaijan. Furthermore, this is the first multicenter study in Turkey and Azerbaijan to identify several major clusters belonging to European clones I and II of A. baumannii.     

Colonization pressure as a risk factor for colonization by multiresistant Acinetobacter spp and carbapenem-resistant Pseudomonas aeruginosa in an intensive care unit

OBJECTIVE:

To determine factors associated with colonization by carbapenem-resistant Pseudomonas aeruginosa and multiresistant Acinetobacter spp.

METHODS:

Surveillance cultures were collected from patients admitted to the intensive care unit at admission, on the third day after admission and weekly until discharge. The outcome was colonization by these pathogens. Two interventions were implemented: education and the introduction of alcohol rubs. Compliance with hand hygiene, colonization pressure, colonization at admission and risk factors for colonization were evaluated.

RESULTS:

The probability of becoming colonized increased during the study. The incidence density of colonization by carbapenem-resistant P. aeruginosa and multiresistant Acinetobacter spp. and colonization pressure were different between periods, increasing gradually throughout the study. The increase in colonization pressure was due to patients already colonized at admission. The APACHE II score, colonization pressure in the week before the outcome and male gender were independent risk factors for colonization. Every 1% increase in colonization pressure led to a 2% increase in the risk of being colonized.

CONCLUSION:

Colonization pressure is a risk factor for carbapenem-resistant P. aeruginosa and multiresistant Acinetobacter spp. colonization. When this pressure reaches critical levels, efforts primarily aimed at hand hygiene may not be sufficient to prevent transmission.     

Clonal relationship and the association of the ST218 strain harboring blaOXA-72 gene to mortality in carbapenem-resistant Acinetobacter baumannii bacteremia

Background/purpose

In 2017, the World Health Organization categorized carbapenem-resistant Acinetobacter baumannii (CRAB) as a priority 1, critical antibiotic-resistant bacteria. This study analyzed the clinical outcomes and investigated the molecular epidemiology of CRAB bacteremia in a medical center in Northern Taiwan.

Further increases in carbapenem-, amikacin-, and fluoroquinolone-resistant isolates of Acinetobacter spp. and P. aeruginosa in Korea: KONSAR study 2009

Purpose

The increasing prevalence of antimicrobial resistant bacteria has become a serious worldwide problem. The aim of this study was to analyze antimicrobial resistance data generated in 2009 by hospitals and commercial laboratories participating in the Korean Nationwide Surveillance of Antimicrobial Resistance program.

Materials and Methods

Susceptibility data were collected from 24 hospitals and two commercial laboratories. In the analysis, resistance did not include intermediate susceptibility. Duplicate isolates were excluded from the analysis of hospital isolates, but not from the commercial laboratory isolates.

Results

Among the hospital isolates, methicillin-resistant Staphylococcus aureus, penicillin G-non-susceptible Streptococcus pneumoniae based on meningitis breakpoint, and ampicillin-resistant Enterococcus faecium remained highly prevalent. The proportion of vancomycin-resistant E. faecium gradually increased to 29%. Ceftazidime-resistant Escherichia coli and Klebsiella pneumoniae increased to 17% and 33%, respectively, and fluoroquinolone-resistant K. pneumoniae, Acinetobacter spp. and Pseudomonas aeruginosa increased to 33%, 67% and 39%, respectively. Amikacin-resistant Acinetobacter spp. increased to 48%. Imipenem-resistant Acinetobacter spp. and P. aeruginosa increased to 51% and 26%, respectively. Higher resistance rates were observed in intensive care unit (ICU) isolates than in non-ICU isolates among the isolates from hospitals. Resistance rates were higher in hospital isolates than in clinic isolates among the isolates from commercial laboratories.

Conclusion

Among the hospital isolates, ceftazidime-resistant K. pneumoniae and fluoroquinolone-resistant K. pneumoniae, Acinetobacter spp., and P. aeruginosa further increased. The increase in imipenem resistance was slight in P. aeruginosa, but drastic in Acinetobacter spp. The problematic antimicrobial-organism combinations were much more prevalent among ICU isolates.     

Inhaled Colistin for Treatment of Pneumonia due to Colistin-Only-Susceptible Acinetobacter baumannii

Purpose

Colistin is used for the treatment of pneumonia associated with multidrug-resistant Acinetobacter baumannii and Pseudomonas aeruginosa. However, the best route of administration and dosage is not known. We report our experience with aerosolized colistin in twelve patients with pneumonia caused by colistin-only-susceptible (COS) A. baumannii.

Materials and Methods

We retrospectively reviewed patients'' medical records who were treated with aerosolized colistin for the treatment of pneumonia.

Results

Ten patients were treated only with aerosolized colistin inhalation and two patients received a 3-day course intravenous colistin, and then switched to colistin inhalation therapy. The median duration of aerosolized colistin therapy was 17 days (5-31 days). Four patients were treated only with aerosolized colistin, whereas 4 patients received concomitant glycopeptides, and 4 received concomitant levofloxacin or cefoperazone/sulbactam. At the end of the therapy, the clinical response rate and bacteriological clearance rate was 83% and 50%, respectively. Colistin-resistant strains were isolated from 3 patients after aerosolized colistin therapy; however, all of them showed favorable clinical response. The median interval between inhalation therapy and resistance was 7 days (range 5-19 days). Acute kidney injury developed in 3 patients. Two patients experienced Clostridium difficile associated diarrhea. One patient developed fever and skin rash after aerosolized colistin therapy. No patient developed neurotoxicity or bronchospasm.

Conclusion

Colistin inhalation therapy is deemed tolerable and safe, and could be beneficial as an adjuctive therapy for the management of pneumonia due to COS A. baumannii. However, the potential development of colistin resistance cannot be overlooked.     

Metallo-��-Lactamase-Producing Pseudomonas spp. in Korea: High Prevalence of Isolates with VIM-2 Type and Emergence of Isolates with IMP-1 Type

Purpose

Two Korean nationwide studies showed that metallo-β-lactamases (MBLs)-producing-Pseudomonas spp. are not rare. The aim of this study was to assess the trends of MBL-producing isolates among imipenem-resistant isolates of Pseudomonas spp.

Materials and Methods

Imipenem-resistant clinical isolates were collected from 23 hospitals and one commercial laboratory participating in the KONSAR program in 2005. Polymerase chain reaction (PCR) was used to detect MBL genes.

Results

Alleles of MBL genes were detected in 10.8% of 415 Pseudomonas aeruginosa and 66.7% of 12 P. putida isolates from 18 of 24 hospitals/laboratory. Among the 14 IMP-1-like and 39 VIM-2-like MBLs, emergence of IMP-6 was detected for the first time.

Conclusion

Prevalence of MBL-producing P. aeruginosa has not significantly increased, but IMP-6 emerged in P. aeruginosa.     

OXA β-Lactamases

SUMMARY

The OXA β-lactamases were among the earliest β-lactamases detected; however, these molecular class D β-lactamases were originally relatively rare and always plasmid mediated. They had a substrate profile limited to the penicillins, but some became able to confer resistance to cephalosporins. From the 1980s onwards, isolates of Acinetobacter baumannii that were resistant to the carbapenems emerged, manifested by plasmid-encoded β-lactamases (OXA-23, OXA-40, and OXA-58) categorized as OXA enzymes because of their sequence similarity to earlier OXA β-lactamases. It was soon found that every A. baumannii strain possessed a chromosomally encoded OXA β-lactamase (OXA-51-like), some of which could confer resistance to carbapenems when the genetic environment around the gene promoted its expression. Similarly, Acinetobacter species closely related to A. baumannii also possessed their own chromosomally encoded OXA β-lactamases; some could be transferred to A. baumannii, and they formed the basis of transferable carbapenem resistance in this species. In some cases, the carbapenem-resistant OXA β-lactamases (OXA-48) have migrated into the Enterobacteriaceae and are becoming a significant cause of carbapenem resistance. The emergence of OXA enzymes that can confer resistance to carbapenems, particularly in A. baumannii, has transformed these β-lactamases from a minor hindrance into a major problem set to demote the clinical efficacy of the carbapenems.     

Metallo-beta-lactamases among imipenem-resistant Pseudomonas aeruginosa in a Brazilian university hospital

INTRODUCTION:

Imipenem‐resistant Pseudomonas aeruginosa resulting from metallo‐β‐lactamases has been reported to be an important cause of nosocomial infection and is a critical therapeutic problem worldwide, especially in the case of bacteremia.

OBJECTIVES:

To determine the frequency of metallo‐β‐lactamases among imipenem‐resistant Pseudomonas aeruginosa isolates and to compare methods of phenotypic and molecular detection.

METHODS:

During 2006, 69 imipenem‐resistant Pseudomonas aeruginosa samples were isolated from blood and tested for metallo‐β‐lactamase production using phenotypic methods. Minimal Inhibitory Concentratrions (MIC) (µg/mL) was determined with commercial microdilution panels. Pulsed Field Gel Electrophoresis (PFGE) was performed among metallo‐β‐lactamase producers.

RESULTS:

Of all the blood isolates, 34.5% were found to be imipenem‐resistant Pseudomonas aeruginosa. Positive phenotypic tests for metallo‐β‐lactamases ranged from 28%‐77%, and Polymerase Chain Reaction (PCR) were positive in 30% (of note, 81% of those samples were bla_SPM‐1 and 19% were bla_VIM‐2). Ethylenediamine tetracetic acid (EDTA) combinations for the detected enzymes had low kappa values; thus, care should be taken when use it as a phenotypic indicator of MBL. Despite a very resistant antibiogram, four isolates demonstrated the worrisome finding of a colistin MIC in the resistant range. PFGE showed a clonal pattern.

CONCLUSION:

Metallo‐β‐lactamases among imipenem‐resistant Pseudomonas aeruginosa were detected in 30.4% of imipenem‐resistant Pseudomonas aeruginosa isolates. This number might have been higher if other genes were included. SPM‐1 was the predominant enzyme found. Phenotypic tests with low kappa values could be misleading when testing for metallo‐β‐lactamases. Polymerase Chain Reaction detection remains the gold standard.     

The prevalence of aminoglycoside-modifying enzyme genes (aac (6')-I, aac (6')-II, ant (2")-I, aph (3')-VI) in Pseudomonas aeruginosa

INTRODUCTION:

Pseudomonas aeruginosa (P. aeruginosa) is one of the primary opportunistic pathogens responsible for nosocomial infections. Aminoglycosides are an important component of antipseudomonal chemotherapy. The inactivation of drugs by modifying enzymes is the most common mechanism of aminoglycoside resistance.

OBJECTIVES:

The inactivation of aminoglycosides by modifying enzymes is the primary resistance mechanism employed by P. aeruginosa. The aim of the present study was to investigate the occurrence of aminoglycoside resistance and the prevalence of four important modifying enzyme genes (aac (6′)-I, aac (6′)-II, ant (2″)-I, aph (3′)-VI) in P. aeruginosa in Iran.

METHODS:

A total of 250 clinical isolates of P. aeruginosa were collected from several hospitals in seven cities in Iran. Antimicrobial susceptibility tests (using the disk diffusion method and E-tests) were performed for all 250 isolates. In addition, all isolates were screened for the presence of modifying enzyme genes by polymerase chain reaction.

RESULTS:

The resistance rates, as determined by the disk diffusion method, were as follows: gentamicin 43%, tobramycin 38%, and amikacin 24%. Of the genes examined, aac (6′)-II (36%) was the most frequently identified gene in phenotypic resistant isolates, followed by ant (2″)-I, aph (3′)-VI, and aac (6′)-I.

CONCLUSIONS:

Aminoglycoside resistance in P. aeruginosa remains a significant problem in Iran. Therefore, there is considerable local surveillance of aminoglycoside resistance.     

Molecular Epidemiology of Multidrug-Resistant Acinetobacter baumannii in a Tertiary Care Hospital in Naples,Italy, Shows the Emergence of a Novel Epidemic Clone

The molecular epidemiology of multidrug-resistant Acinetobacter baumannii was investigated in two intensive care units of the V. Monaldi university hospital in Naples, Italy, from May 2006 to December 2007. Genotype analysis by pulsed-field gel electrophoresis (PFGE), trilocus sequence-based typing (3LST), and multilocus sequence typing (MLST) of A. baumannii isolates from 71 patients identified two distinct genotypes, one assigned to PFGE group A, 3LST group 1, and ST2 in 14 patients and the other to PFGE group B, 3LST group 6, and ST78 in 71 patients, that we named ST2/A and ST78/B, respectively. Of these, ST2/A corresponded to European clone II identified in the same hospital during 2003 and 2004; ST78/B was a novel genotype that was isolated for the first time in May 2006 but became prevalent during 2007. The ST78/B profile was also identified in five patients from two additional hospitals in Naples during 2007. The ST2/A and ST78/B isolates were resistant to all antimicrobials tested, including carbapenems, but were susceptible to colistin. Both ST2/A and ST78/B isolates possessed a plasmid-borne carbapenem-hydrolyzing oxacillinase gene, bla_OXA-58, flanked by ISAba2 and ISAba3 elements at the 5′ and 3′ ends, respectively. The selection of the novel ST78/B A. baumannii clone might have been favored by the acquisition of the bla_OXA-58 gene.Acinetobacter baumannii is an emerging opportunistic nosocomial pathogen, with increasing prevalence worldwide, responsible for a variety of nosocomial infections, especially in intensive care unit (ICU) patients (10, 15). Several hospital outbreaks caused by the selection of multiresistant A. baumannii clones have been described in Europe and worldwide (1, 10, 15, 27). Genotypic characterization of epidemic A. baumannii isolates through amplified fragment length polymorphism analysis has identified clusters of highly similar strains, which were assumed to represent distinct clonal lineages and were defined as European clones I, II, and III (9, 24). Similarly, three distinct groups were recently identified among A. baumannii isolates from five different countries by sequence-based typing (ST): group 1, corresponding to European clone II; group 2, corresponding to European clone I; and group 3, corresponding to European clone III (22). Moreover, epidemics caused by A. baumannii genotypes assigned to novel ST groups 4 and 5 have been recently described in different Greek and Turkish cities (11). The majority of the outbreaks occurring in Europe were caused by carbapenem-resistant strains that carried the bla_OXA-58 gene or a distinct carbapenem-hydrolyzing oxacillinase (CHDL) gene (4, 8, 11-13, 16, 20, 27, 28).We have previously reported the occurrence of two sequential outbreaks from August 1999 to February 2001 and from January 2002 to December 2002 along with the emergence of carbapenem-resistant A. baumannii in the ICU of the Federico II university hospitals in Naples, Italy, during 2002 (26). More recently, we have shown that the same epidemic A. baumannii clone isolated during 2002 was responsible for a large and sustained outbreak in the V. Monaldi tertiary care teaching hospital of Naples between June 2003 and June 2004 (25). An increase in the number of cases of A. baumannii infection was observed after 2 years in the V. Monaldi hospital. The objectives of the present study were (i) to investigate the molecular epidemiology of A. baumannii in the V. Monaldi hospital, (ii) to study the genetic characteristics of A. baumannii isolates responsible for the epidemic, and (iii) to analyze the antimicrobial susceptibilities of the A. baumannii isolates and their mechanisms of resistance.     

Characterization of Epidemiologically Unrelated Acinetobacter baumannii Isolates from Four Continents by Use of Multilocus Sequence Typing,Pulsed-Field Gel Electrophoresis,and Sequence-Based Typing of blaOXA-51-like Genes

This study used a diverse collection of epidemiologically unrelated Acinetobacter baumannii isolates to compare the robustness of a multilocus sequence typing (MLST) scheme, based on conserved regions of seven housekeeping genes, gltA, gdhB, recA, cpn60, rpoD, gyrB, and gpi, with that of sequence-based typing of bla_OXA-51-like genes (SBT-bla_OXA-51-like genes). The data obtained by analysis of MLST and SBT-bla_OXA-51-like genes were compared to the data generated by pulsed-field gel electrophoresis (PFGE). The topologies of the phylogenetic trees generated for the gyrB and gpi genes showed evidence of recombination and were inconsistent with those of the trees generated for the other five genes. MLST identified 24 sequence types (STs), of which 19 were novel, and 5 novel alleles. Clonality was demonstrated by eBURST analysis and standardized index of association values of >1 (P < 0.001). MLST data revealed that all isolates harboring the major bla_OXA-51-like alleles OXA-66, OXA-69, and OXA-71 fell within the three major European clonal lineages. However, the MLST data were not always in concordance with the PFGE data, and some isolates containing the same bla_OXA-51-like allele demonstrated <50% relatedness by PFGE. It was concluded that the gyrB and gpi genes are not good candidates for use in MLST analysis and that a SBT-bla_OXA-51-like gene scheme produced results comparable to those produced by MLST for the identification of the major epidemic lineages, with the advantage of having a significantly reduced sequencing cost and time. It is proposed that studies of A. baumannii epidemiology could involve initial screening of bla_OXA-51-like alleles to identify isolates belonging to major epidemic lineages, followed by MLST analysis to categorize isolates from common lineages, with PFGE being reserved for fine-scale typing.Acinetobacter baumannii is a Gram-negative bacterium that causes serious nosocomial infections, especially in critical care units (13, 14, 42). Several outbreaks have been caused by multidrug-resistant (MDR) strains of A. baumannii (23, 33, 43), and the rate of resistance to carbapenems, which have been the antibiotics of choice to treat infections caused by this pathogen, has increased considerably over the last decade (4, 32, 42, 46). In addition, the prevalence of A. baumannii in hospitals has increased worldwide (3, 25, 28, 29, 47), and, therefore, finding suitable molecular typing methods for A. baumannii is essential for epidemiological investigations and infection control studies. Many genomic typing methods have been used, including ribotyping (36), infrequent-restriction-site analysis (48), repetitive extragenic palindromic sequence-based PCR (rep-PCR) (18), random amplified polymorphic DNA (RAPD) analysis (21), amplified fragment length polymorphism (AFLP) analysis (39), and multilocus PCR and electrospray ionization mass spectrometry (PCR/ESI-MS) (6). Pulsed-field gel electrophoresis (PFGE) is still considered the “gold standard” for the typing of bacterial isolates (36), but it has drawbacks when its comes to interchanging data among laboratories for comparison purposes (35) and may lose its discriminatory power when isolates from geographically diverse areas are analyzed.Multilocus sequence typing (MLST) schemes, which use several housekeeping genes, have already been used to type many pathogenic bacteria (15, 17, 20, 40), including A. baumannii (1, 31, 44), and MLST is emerging as an alternative to PFGE. MLST is used mainly for global epidemiology studies, but it has also been used successfully for short-term investigation of an outbreak of meningococcal disease (11). Although MLST has many advantages over other molecular typing methods, many questions remain to be answered, including whether several loci are required to obtain a robust scheme and whether the criteria for the selection of the housekeeping genes are sufficiently reliable to reveal the population structure of the strains analyzed.bla_OXA-51-like genes are unique to A. baumannii and may be used as markers for identification of this species (16). They have also successfully been used as one of three loci in a PCR-based typing scheme that is able to assign isolates of A. baumannii to sequence groups (SGs) that appear to correlate with the major epidemic lineages within the species (41). This raises the question of whether the bla_OXA-51-like genes themselves could be utilized in a typing scheme. Accordingly, the aim of this work was to investigate the robustness of MLST in categorizing epidemiologically unrelated A. baumannii isolates from four continents and to evaluate the use of variations within the intrinsic bla_OXA-51-like gene as a typing tool comparable to MLST.     

Association of biofilm production with multidrug resistance among clinical isolates of Acinetobacter baumannii and Pseudomonas aeruginosa from intensive care unit

Background and Aims:

Given choice, bacteria prefer a community-based, surface-bound colony to an individual existence. The inclination for bacteria to become surface bound is so ubiquitous in diverse ecosystems that it suggests a strong survival strategy and selective advantage for surface dwellers over their free-ranging counterparts. Virtually any surface, biotic or abiotic (animal, mineral, or vegetable) is suitable for bacterial colonization and biofilm formation. Thus, a biofilm is “a functional consortium of microorganisms organized within an extensive exopolymeric matrix.”

Materials and Methods:

The present study was undertaken to detect biofilm production from the repertoire stocks of Acinetobacter baumannii (A. baumannii) and Pseudomonas aeruginosa (P. aeruginosa) obtained from clinical specimens. The tube method was performed to qualitatively detect biofilm production.

Results:

A total of 109 isolates of both organisms were included in the study, out of which 42% (46/109) isolates showed biofilm detection. Among the biofilm producers, 57% of P. aeruginosa and 73% of A. baumannii showed multidrug resistance (MDR) pattern which was statistically significant in comparison to nonbiofilm producers (P < 0.001).

Conclusion:

To the best of our knowledge, this is the only study to have tested the biofilm production in both P. aeruginosa and A. baumannii in a single study. Biofilm production and MDR pattern were found to be significantly higher in A. baumannii than P. aeruginosa. Antibiotic resistance was significantly higher among biofilm producing P. aeruginosa than non producers. Similarly, antibiotic resistance was significantly higher among biofilm producing A. baumannii than non producers.     

Nasal carriage of multi-drug resistant Staphylococcus aureus in healthy inhabitants of Amassoma in Niger delta region of Nigeria

Background

Nasal Staphylococcus aureus is a major source of community and hospital associated staphylococcal infections. This study determined the prevalence of nasal S. aureus isolates and investigated their antimicrobial resistance profile in healthy volunteers.

Methods

Nasal specimens of healthy volunteers in Amassoma were cultured and screened for S. aureus using standard microbiological protocols and their antibiotic profile susceptibility was investigated using disc diffusion and agar dilution techniques.

Results

A total of 40 (33.3%) S. aureus isolates were obtained from 120 nares specimens screened. Twenty three (57.5%) and 17 (42.5%) of the isolates were from university students and villagers respectively. The isolates showed an overall 75% resistance to ampicillin, 52.5% to doxycycline, 47.5% to chloramphenicol, 35% to erythromycin and 32.5% to cotrimoxazole; with 27.5% methicillin resistant. No isolate was resistant to gentamicin while few isolates were resistant to cefuroxime (2.5%), augmentin (5.0%), ciprofloxacin (10.0%), ofloxacin (10.0%) and vancomycin (7.5%). Twenty one (52.5%) of all the isolates were multi-drug resistant, ten (47.6%) of which were methicillin resistant Staphylococcus aureus (MRSA) and only 3 (7.5%) were fully susceptible to all the tested antimicrobial drugs.

Conclusions

The observation calls for strategies to prevent their spread to more vulnerable populations where the consequences of their infections can be severe.     

Prevalence of carbapenem-hydrolyzing class D β-lactamase genes in Acinetobacter spp. isolates in China

In order to assess the prevalence of carbapenem-hydrolyzing class D β-lactamase genes in Acinetobacter spp. isolates in China, we conducted a polymerase chain reaction (PCR)-based surveillance of OXA-type β-lactamase gene clusters for a total of 2,880 Acinetobacter spp. isolates collected from 23 Chinese provinces. All isolates were tested for susceptibility to 12 antimicrobial agents and showed high rates of resistance to all these agents except minocycline. We also found that the vast majority of carbapenem-resistant Acinetobacter spp. were OXA-23-like-producing isolates, predominantly Acinetobacter baumannii isolates. Besides, bla _OXA-58-like and bla _OXA-24-like genes were detected in 32 and 11 isolates, respectively, involving many provinces throughout China. Furthermore, these two carbapenem-resistance determinants were located on transferable plasmids in most cases, indicating an emerging threat for both OXA-58-like- and OXA-24-like-producing Acinetobacter spp. isolates in China. Interestingly, a novel homologue of the bla _OXA-143 gene was identified in a susceptible Acinetobacter pittii isolate. Overall, these observations suggest that the bla _OXA-23-harboring A. baumannii isolates are the most frequent carbapenem-resistant Acinetobacter spp. in China, and the bla _OXA-24-like and bla _OXA-58-like genes have emerged as potential threats of hospital outbreaks of multidrug-resistant Acinetobacter spp.     

Risk Factors and Molecular Epidemiology of Community-Onset Extended-Spectrum β-Lactamase-Producing Escherichia coli Bacteremia

Purpose

Inadequate empirical therapy for severe infections caused by extended-spectrum β-lactamase-producing Escherichia coli (ESBLEC) is associated with poor outcomes. This study was designed to investigate risk factors for community-onset ESBLEC bacteremia at admission to a tertiary care hospital.

Materials and Methods

A case-control study was performed that included all episodes of ESBLEC bacteremia in the outpatient department or within 48 hours of admission from January 2005 to March 2009. Data on predisposing factors were collected. The molecular epidemiology of ESBLEC clinical isolates was also determined.

Results

Among 25281 blood cultures, 60 episodes of ESBLEC bacteremia were studied, which accounted for 7% of all E. coli bacteremia at admission. Healthcare-associated infection [odds ratio (OR), 8.3; 95% confidence interval (CI), 2.4-28.7; p=0.001], malignancy (OR, 4.6; 95% CI, 1.3-16.3; p=0.018), urinary tract infection (OR, 139.1; 95% CI, 24.6-788.2; p<0.001), hepatobiliary infection (OR, 79.1; 95% CI, 13.5-463.8; p<0.001), third generation cephalosporin usage during preceding 3 months (OR, 16.4; 95% CI, 2.0-131.8; p=0.008), and severe sepsis/septic shock (OR, 73.7; 95% CI, 12.4-438.5; p<0.001) were determined as independent risk factors for community-onset ESBLEC bacteremia. The most common extended-spectrum β-lactamase (ESBL) gene identified was bla_CTX-M-15 (n=31) followed by bla_CTX-M-14 (n=23).

Conclusion

The most common types of ESBLs in E. coli causing community-onset bacteremia were CTX-M-15 and CTX-M-14 in Korea. By result of decision tree analysis, the empirical use of carbapenems is suggested only for patients with severe sepsis/septic shock, hepatobiliary infection, or healthcare-associated urinary tract infection.     

Clinical review of endogenous endophthalmitis in Korea: a 14-year review of culture positive cases of two large hospitals

Purpose

To identify the clinical features and outcomes of endogenous endophthalmitis in Korea.

Materials and Methods

We reviewed 18 patients with endogenous endophthalmitis at 2 Korean hospitals, treated over a 14 year period between January 1993 and December 2006.

Results

The comorbidities observed in these cases were diabetes mellitus and liver cirrhosis. The most common pathogens, which were found in 7 patients each (38.9%), were Klebsiella pneumonia and Pseudomonas aeruginosa. All patients were treated with systemic antibiotics and fortified topical antibiotics. A surgical approach including vitrectomy was performed in 9 cases (50.0%). The prognosis was generally poor, and visual acuity improved slightly in 6 patients (33.3%).

Conclusion

In this study, diabetes mellitus and Klebsiella pneumonia showed a close relationship with endogenous endophthalmitis, respectively. Endogenous endophthalmitis is a serious risk to sight and careful attention to establishing the diagnosis and management may decrease the ocular morbidity.     

The modified Hodge test is a useful tool for ruling out klebsiella pneumoniae carbapenemase

OBJECTIVE:

Enterobacteriaceae bacteria harboring Klebsiella pneumoniae carbapenemase are a serious worldwide threat. The molecular identification of these pathogens is not routine in Brazilian hospitals, and a rapid phenotypic screening test is desirable. This study aims to evaluate the modified Hodge test as a phenotypic screening test for Klebsiella pneumoniae carbapenemase.

METHOD:

From April 2009 to July 2011, all Enterobacteriaceae bacteria that were not susceptible to ertapenem according to Vitek2 analysis were analyzed with the modified Hodge test. All positive isolates and a random subset of negative isolates were also assayed for the presence of bla_KPC. Isolates that were positive in modified Hodge tests were sub-classified as true-positives (E. coli touched the ertapenem disk) or inconclusive (distortion of the inhibition zone of E. coli, but growth did not reach the ertapenem disk). Negative results were defined as samples with no distortion of the inhibition zone around the ertapenem disk.

RESULTS:

Among the 1521 isolates of Enterobacteriaceae bacteria that were not susceptible to ertapenem, 30% were positive for bla_KPC, and 35% were positive according to the modified Hodge test (81% specificity). Under the proposed sub-classification, true positives showed a 98% agreement with the bla_KPC results. The negative predictive value of the modified Hodge test for detection was 100%. KPC producers showed high antimicrobial resistance rates, but 90% and 77% of these isolates were susceptible to aminoglycoside and tigecycline, respectively.

CONCLUSION:

Standardizing the modified Hodge test interpretation may improve the specificity of KPC detection. In this study, negative test results ruled out 100% of the isolates harboring Klebsiella pneumoniae carbapenemase-2. The test may therefore be regarded as a good epidemiological tool.