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Etiology of blood culture isolates among patients in a multidisciplinary teaching hospital in Kuala Lumpur

Karunakaran R, Raja NS, Ng KP, Navaratnam P

J Microbiol Immunol Infect, 2007 Oct;40(5):432-7.
PMID: 17932604

Bloodstream infections are an important cause of morbidity and mortality among hospitalized patients and the surveillance of etiological agents in these infections is important for their prevention and treatment. Data on common organisms isolated from blood cultures from Malaysia are limited, and our aim was to identify the common bloodstream isolates in hospitalized patients at the University of Malaya Medical Centre (UMMC), Kuala Lumpur, Malaysia.

Matched MeSH terms: beta-Lactamases/biosynthesis
Comparison of three different methods for the presumptive detection of ESBL production in ceftazidime-resistant strains of Klebsiella pneumoniae

Palasubramaniam S, Parasakthi N

Malays J Pathol, 2001 Dec;23(2):73-8.
PMID: 12166595

Twenty-eight (28) strains of ceftazidime-resistant Klebsiella pneumoniae were isolated from blood cultures of in-patients from University Hospital, Kuala Lumpur between March 1995 and August 1996. Three methods were used to detect the production of ESBL enzymes by these strains. These three methods include the double-disc synergy test (DDST), inhibitor-potentiated disc-diffusion test (IPDD) and the E-test ESBL method. All strains could be identified as ESBL producers using the DDST method by a minimum of two beta-lactams and these included either a combination of ceftazidime and ceftriaxone with clavulanate respectively or cefotaxime and aztreonam with clavulanate respectively. Similarly using a combination of either cefotaxime and ceftriaxone with clavulanate or ceftriaxone and aztreonam with clavulanate respectively would have detected all strains as ESBL producers. The IPDD method could also detect for ESBL activity based on combinations of beta-lactam antibiotics with clavulanate respectively. All combinations of beta-lactam antibiotics could detect for ESBL activity in all the strains except a combination of either ceftazidime and aztreonam or cefotaxime and ceftriaxone with clavulanate respectively. The E-Test method using ceftazidime alone and in combination with clavulanate was found to be the most effective method in the presumptive identification of ESBL activity in all the strains.

Matched MeSH terms: beta-Lactamases/biosynthesis*
Infections due to beta-lactamase-producing Neisseria gonorrhoeae at the University Hospital Medical Center, Kuala Lumpur, Malaysia

Ismail R

Sex Transm Dis, 1987 4 1;14(2):113-5.
PMID: 3112967

The incidence of infections due to beta-lactamase-producing Neisseria gonorrhoeae is increasing in many parts of the world. An epidemiologic survey of infections caused by beta-lactamase-producing strains of N. gonorrhoeae at the University Hospital, Kuala Lumpur, from February 1977 to December 1985 (106 months) showed that the incidence rose from 4.8% (two cases) in 1977 to 49.4% (39 cases) by the end of 1985. The highest incidence of gonococcal infections was found to be in the group aged 20-39 years; the male-to-female ratio was 1.55:1. The mean inhibitory concentrations of benzylpenicillin were 0.12 microgram/ml for non-beta-lactamase-producing strains and 16 micrograms/ml for isolates of N. gonorrhoeae that produce beta-lactamase.

Matched MeSH terms: beta-Lactamases/biosynthesis*
Detection of VIM-2-, IMP-1- and NDM-1-producing multidrug-resistant Pseudomonas aeruginosa in Malaysia

Liew SM, Rajasekaram G, Puthucheary SD, Chua KH

J Glob Antimicrob Resist, 2018 06;13:271-273.
PMID: 29432937 DOI: 10.1016/j.jgar.2018.01.026

OBJECTIVES: The increasing incidence of carbapenem-resistant Pseudomonas aeruginosa along with the discovery of novel metallo-β-lactamases (MBLs) is of concern. In this study, the isolation of MBL-producing P. aeruginosa clinical strains in Malaysia was investigated.
METHODS: A total of 53 P. aeruginosa clinical strains were isolated from different patients in Sultanah Aminah Hospital (Johor Bahru, Malaysia) in 2015. Antimicrobial susceptibility testing was performed, and minimum inhibitory concentrations (MICs) of imipenem and meropenem were determined by Etest. Carbapenem-resistant strains were screened for MBL production by the imipenem-ethylene diamine tetra-acetic acid (IMP-EDTA) double-disk synergy test, MBL imipenem/imipenem-inhibitor (IP/IPI) Etest and PCR. Multilocus sequence typing (MLST) analysis was performed for genotyping of the isolates.
RESULTS: Among the 53 clinical strains, 3 (5.7%) were identified as MBL-producers. Multidrug resistance was observed in all three strains, and two were resistant to all of the antimicrobials tested. Sequencing analysis confirmed that the three strains harboured carbapenemase genes (blaIMP-1, blaVIM-2 and blaNDM-1 in one isolate each). These multidrug-resistant strains were identified as sequence type 235 (ST235) and ST308.
CONCLUSIONS: The blaIMP-1 and blaNDM-1 genes have not previously been reported in Malaysian P. aeruginosa isolates. The emergence of imipenemase 1 (IMP-1)- and New Delhi metallo-β-lactamase 1 (NDM-1)-producing P. aeruginosa in Malaysia maybe travel-associated.

Matched MeSH terms: beta-Lactamases/biosynthesis
Fulltext Detection and characterization of ESBL-producing Enterobacteriaceae from the gut of subsistence farmers, their livestock, and the surrounding environment in rural Nepal

Subramanya SH, Bairy I, Metok Y, Baral BP, Gautam D, Nayak N

Sci Rep, 2021 01 22;11(1):2091.
PMID: 33483551 DOI: 10.1038/s41598-021-81315-3

The increasing trend of gut colonization by extended-spectrum β-lactamase (ESBL) producing Enterobacterales has been observed in conventional farm animals and their owners. Still, such colonization among domesticated organically fed livestock has not been well studied. This study aimed to determine the gut colonization rate of ESBL-producing Enterobacteriaceae and carbapenemase-producing Enterobacteriaceae (CPE) among rural subsistence farming communities of the Kaski district in Nepal. Rectal swabs collected by systematic random sampling from 128 households of subsistence farming communities were screened for ESBL-producing Enterobacteriaceae and CPE by phenotypic and molecular methods. A total of 357 (57%) ESBL-producing Enterobacteriaceae isolates were obtained from 626 specimens, which included 97 ESBL-producing Enterobacteriaceae (75.8%) from 128 adult humans, 101 (79.5%) from 127 of their children, 51 (47.7%) from 107 cattle, 26 (51%) from 51 goats, 30 (34.9%) from 86 poultry and 52 (42%) from 127 environmental samples. No CPE was isolated from any of the samples. blaCTX-M-15 was the most predominant gene found in animal (86.8%) and human (80.5%) isolates. Out of 308 Escherichia coli isolates, 16 human and two poultry isolates were positive for ST131 and were of clade C. Among non-cephalosporin antibiotics, the resistance rates were observed slightly higher in tetracycline and ciprofloxacin among all study subjects. This is the first one-health study in Nepal, demonstrating the high rate of CTX-M-15 type ESBL-producing Enterobacteriaceae among gut flora of subsistence-based farming communities. Gut colonization by E. coli ST131 clade C among healthy farmers and poultry birds is a consequential public health concern.

Matched MeSH terms: beta-Lactamases/biosynthesis*
Detection of betalactamase producing bacterial genes and their clinical features

Hashim RB, Husin S, Rahman MM

Pak J Biol Sci, 2011 Jan 01;14(1):41-6.
PMID: 21913496

The present study was aimed to identify the gene of drug resistance betalactamase producing bacteria and clinical features of the infected patients at Hospital University Kebangsaan Malaysia. Blood samples from the patients were collected, processed and betalactamase producing drug resistance bacteria were identified by antibiotic sensitivity testing. Genes of the drug resistance bacteria were detected and characterized by polymerase chain reaction. A total of 34 isolates of drug resistance Betalactamase producing E. coli and Klebsiella spp. were isolated from 2,502 patients. Most common drug resistance gene TEM was found in 50% of the isolates. 11% was found positive for both TEM and SHV. Next 11% of the isolates expressed only SHV genes. Clinical features of the patients were recorded from where the bacteria isolated. Regarding community affiliations 70.5% of the infected patients were Malay 17.6% were Indian and 11.7% were Chinese. Majority of the patients has an underlying pre-morbid condition as reflected by their diagnosis. Better infection control and hygiene in hospitals, plus controlled and prudent use of antibiotics, is required to minimize the impact of drug resistance betalactamase producing bacteria and the spread of infections.

Matched MeSH terms: beta-Lactamases/biosynthesis*
Imipenem-resistance in Klebsiella pneumoniae in Malaysia due to loss of OmpK36 outer membrane protein coupled with AmpC hyperproduction

Palasubramaniam S, Karunakaran R, Gin GG, Muniandy S, Parasakthi N

Int J Infect Dis, 2007 Sep;11(5):472-4.
PMID: 17337225
Matched MeSH terms: beta-Lactamases/biosynthesis*
Rapid detection of ESBL-producing Klebsiella pneumoniae in blood cultures by fluorescent in-situ hybridization

Palasubramaniam S, Muniandy S, Navaratnam P

J Microbiol Methods, 2008 Jan;72(1):107-9.
PMID: 18054098

Multi-resistant Enterobacteriaceae pose a serious threat of hospital acquired infections and their rapid identification is important for better clinical outcome. This study describes the rapid identification of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae of the sulphydryl variable-type by fluorescent in-situ hybridization. The method which rapidly identifies the target genes within 1 h could be a potentially rapid bacterial diagnostic tool.

Matched MeSH terms: beta-Lactamases/biosynthesis
Fulltext First Report of Extended-Spectrum β-Lactamases TEM-116 and OXA-10 in Clinical Isolates of Alcaligenes Species from Kuala Lumpur, Malaysia

Puah SM, Puthucheary SD, Chua KH

Jpn J Infect Dis, 2019 Jul 24;72(4):266-269.
PMID: 30918144 DOI: 10.7883/yoken.JJID.2018.031

There is an alarming increase in the prevalence of extended-spectrum β-lactamases (ESBLs) present mainly in Enterobacteriaceae and other nonfermenting gram-negative bacteria, such as Alcaligenes faecalis, which is the only species in that genus that is clinically relevant. We investigated Alcaligenes species from 7 cases (6 inpatients and one outpatient) at our tertiary-care hospital. Four patients had urinary tract infections, and one each had systemic lupus erythematosus, pulmonary stenosis, and diabetic ulcer. All 7 isolates were identified as Alcaligenes spp. based on their 16S rRNA gene sequences, and antibiotic susceptibility was determined using a Vitek 2 system with AST-GN87 cards. All the strains were resistant to cefazolin; 6 were resistant to trimethoprim/sulfamethoxazole; 5 manifested resistance to ampicillin/sulbactam, cefepime, tobramycin, ciprofloxacin, and nitrofurantoin; whereas 5 had multidrug resistance profiles. All the strains (7/7) expressed ESBL activity; PCR screening and sequencing showed evidence of genes blaTEM-116 (7/7) and blaOXA-10 (4/7), and we believe that this is the first report on the presence of TEM-116 and OXA-10 in an Alcaligenes spp. A combination of the 2 genes was present in 4 strains. All 7 strains were found to harbor at least one ESBL gene probably contributing to the drug resistance.

Matched MeSH terms: beta-Lactamases/biosynthesis
Fulltext Integrating multiple genomic technologies to investigate an outbreak of carbapenemase-producing Enterobacter hormaechei

Roberts LW, Harris PNA, Forde BM, Ben Zakour NL, Catchpoole E, Stanton-Cook M, et al.

Nat Commun, 2020 01 24;11(1):466.
PMID: 31980604 DOI: 10.1038/s41467-019-14139-5

Carbapenem-resistant Enterobacteriaceae (CRE) represent an urgent threat to human health. Here we report the application of several complementary whole-genome sequencing (WGS) technologies to characterise a hospital outbreak of blaIMP-4 carbapenemase-producing E. hormaechei. Using Illumina sequencing, we determined that all outbreak strains were sequence type 90 (ST90) and near-identical. Comparison to publicly available data linked all outbreak isolates to a 2013 isolate from the same ward, suggesting an environmental source in the hospital. Using Pacific Biosciences sequencing, we resolved the complete context of the blaIMP-4 gene on a large IncHI2 plasmid carried by all IMP-4-producing strains across different hospitals. Shotgun metagenomic sequencing of environmental samples also found evidence of ST90 E. hormaechei and the IncHI2 plasmid within the hospital plumbing. Finally, Oxford Nanopore sequencing rapidly resolved the true relationship of subsequent isolates to the initial outbreak. Overall, our strategic application of three WGS technologies provided an in-depth analysis of the outbreak.

Matched MeSH terms: beta-Lactamases/biosynthesis*
Analysis of plasmid and chromosomal DNA of multidrug-resistant Salmonella enterica serovar typhi from Asia

Mirza S, Kariuki S, Mamun KZ, Beeching NJ, Hart CA

J Clin Microbiol, 2000 Apr;38(4):1449-52.
PMID: 10747124

Molecular analysis of chromosomal DNA from 193 multidrug-resistant (MDR) Salmonella enterica serovar Typhi isolates from 1990 to 1995 from Pakistan, Kuwait, Malaysia, Bangladesh, and India produced a total of five major different pulsed-field gel electrophoresis (PFGE) patterns. Even within a particular country MDR S. enterica serovar Typhi DNA was found to be in different PFGE groups. Similar self-transferable 98-MDa plasmids belonging to either incompatibility group incHI1 or incHI1/FIIA were implicated in the MDR phenotype in S. enterica serovar Typhi isolates from all the locations except Quetta, Pakistan, where the majority were of incFIA. A total of five different PFGE genotypes with six different plasmids, based on incompatibility and restriction endonuclease analysis groups, were found among these MDR S. enterica serovar Typhi isolates.

Matched MeSH terms: beta-Lactamases/biosynthesis
Resistance to extended-spectrum beta-lactams by the emergence of SHV-12 and the loss of OmpK35 in Klebsiella pneumoniae and Escherichia coli in Malaysia

Palasubramaniam S, Muniandy S, Navaratnam P

J Microbiol Immunol Infect, 2009 Apr;42(2):129-33.
PMID: 19597644

In addition to beta-lactamase production, loss of porins confers resistance to extended-spectrum beta-lactams in Klebsiella pneumoniae and Escherichia coli infection. This study describes the detection of SHV-12 extended-spectrum beta-lactamase (ESBL) subtype and the loss of OmpK35 porin in 4 strains of K. pneumoniae and E. coli.

Matched MeSH terms: beta-Lactamases/biosynthesis*
Extended-spectrum beta-lactam resistance due to AmpC hyperproduction and CMY-2 coupled with the loss of OMPK35 in Malaysian strains of Escherichia coli and Klebsiella pneumoniae

Palasubramaniam S, Subramaniam G, Muniandy S, Parasakthi N

Microb Drug Resist, 2007;13(3):186-90.
PMID: 17949305

In this report, we describe the detection of AmpC and CMY-2 beta-lactamases with the loss of OmpK35 porin among seven sporadic strains of ceftazidime-resistant Klebsiella pneumoniae and ceftazidime-resistant Escherichia coli. Cefoxitin, which was used as a marker of resistance toward 7-alpha-methoxy-cephalosporins, exhibited high minimum inhibitory concentration (MIC) values ranging between 128 microg/ml and >256 microg/ml in all the strains. The presence of hyperproducing AmpC enzymes was indicated by the positive three-dimensional test. Isoelectric focusing (IEF) study confirmed the presence of AmpC enzymes in all the strains. The ampC gene was detected by PCR in all the strains and confirmed by DNA sequencing. Large plasmids in all the strains, ranging from 60 kb to 150 kb in size, most likely encode the ampC gene. Two E. coli strains out of the seven strains showed positive amplification of the bla(CMY-2) gene, an AmpC variant, and was confirmed by DNA sequence analyses. DNA hybridization confirmed the bla(CMY-2) gene to be plasmid-mediated in both of these strains. However, one of these two strains also mediated a chromosomal CMY gene. All the strains showed an absence of OmpK35 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS/PAGE) and was confirmed by western blot analyses using raised polyclonal anti-OmpK35 antiserum. This suggests that, apart from CMY production, absence of OmpK35 porin also contributed to cefoxitin resistance resulting in extended-spectrum beta-lactam resistance among these isolates.

Matched MeSH terms: beta-Lactamases/biosynthesis*
Antibiotic resistance patterns in nosocomial gram-negative bacterial infections in units with heavy antibiotic usage

Ariffin H, Navaratnam P, Kee TK, Balan G

J Trop Pediatr, 2004 Feb;50(1):26-31.
PMID: 14984166

The pattern of antibiotic resistance amongst gram-negative bacteria (GNB) in paediatric units, which have heavy empirical usage of broad-spectrum antibiotics, was studied prospectively over a 6-month period. A total of 200 consecutive, non-duplicate gram-negative isolates were obtained from 109 patients admitted to intensive care and oncology units in two hospitals. The commonest isolates were Klebsiella spp (36.5 per cent) and Pseudomonas (20.0 per cent). The isolates showed lower susceptibility rates to the third-generation cephalosporins (47-62 per cent) compared with cefepime (91 per cent), imipenem (90 per cent) and ciprofloxacin (99 per cent). Fifty-four (52.8 per cent) Klebsiella and Escherichia coli isolates were determined to be extended-spectrum beta-lactamase (ESBL) producing strains. Antibiotics found to be effective against ESBL-producers were imipenem and ciprofloxacin. The high resistance rate amongst GNB to third-generation cephalosporins is a likely consequence of heavy empirical usage of this group of antibiotics. The carbapenems and quinolones remain useful agents in the management of patients admitted to these units.

Matched MeSH terms: beta-Lactamases/biosynthesis
Fulltext Characterization of multidrug resistant ESBL-producing Escherichia coli isolates from hospitals in Malaysia

Lim KT, Yasin R, Yeo CC, Puthucheary S, Thong KL

J Biomed Biotechnol, 2009;2009:165637.
PMID: 19672454 DOI: 10.1155/2009/165637

The emergence of Escherichia coli that produce extended spectrum beta-lactamases (ESBLs) and are multidrug resistant (MDR) poses antibiotic management problems. Forty-seven E. coli isolates from various public hospitals in Malaysia were studied. All isolates were sensitive to imipenem whereas 36 were MDR (resistant to 2 or more classes of antibiotics). PCR detection using gene-specific primers showed that 87.5% of the ESBL-producing E. coli harbored the blaTEM gene. Other ESBL-encoding genes detected were blaOXA, blaSHV, and blaCTX-M. Integron-encoded integrases were detected in 55.3% of isolates, with class 1 integron-encoded intI1 integrase being the majority. Amplification and sequence analysis of the 5'CS region of the integrons showed known antibiotic resistance-encoding gene cassettes of various sizes that were inserted within the respective integrons. Conjugation and transformation experiments indicated that some of the antibiotic resistance genes were likely plasmid-encoded and transmissible. All 47 isolates were subtyped by PFGE and PCR-based fingerprinting using random amplified polymorphic DNA (RAPD), repetitive extragenic palindromes (REPs), and enterobacterial repetitive intergenic consensus (ERIC). These isolates were very diverse and heterogeneous. PFGE, ERIC, and REP-PCR methods were more discriminative than RAPD in subtyping the E. coli isolates.

Matched MeSH terms: beta-Lactamases/biosynthesis*

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