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  1. Lee CL, Ng HF, Ngeow YF, Thaw Z
    J Med Microbiol, 2021 Jul;70(7).
    PMID: 34236301 DOI: 10.1099/jmm.0.001378
    Introduction. Tigecycline is currently acknowledged to be one of the most effective antibiotics against infections caused by Mycobacteroides abscessus.Gap statement. The genetic determinants of tigecycline resistance in M. abscessus are not well understood.Aim. In this study, we characterized a tigecycline-resistant M. abscessus mutant, designated CL7, to identify the potential resistance mechanism.Methodology. CL7 was characterized using antimicrobial susceptibility testing, whole-genome sequencing, PCR and RT-qPCR. For biological verification, gene overexpression assays were carried out.Results. Whole-genome sequencing and the subsequent gene overexpression assays showed that CL7 harboured a stop-gain mutation in MAB_3543 c, which may be responsible for the tigecycline resistance phenotype. This gene encodes an orthologue of SigH, which is involved in the positive regulation of physiological stress response and is negatively regulated by the RshA anti-sigma factor in Mycobacterium tuberculosis. We hypothesized that the MAB_3543 c mutation may disrupt the interaction between SigH and RshA (MAB_3542 c). RT-qPCR analyses revealed the upregulation of MAB_3543 c and other key stress response genes, which has previously been shown to be a hallmark of SigH-RshA bond disruption and tigecycline resistance.Conclusion. The MAB_3543c mutation may represent a novel determinant of tigecycline resistance in M. abscessus. The findings of this study will hopefully contribute to our knowledge of potential tigecycline resistance mechanisms in M. abscessus, which may lead to better diagnostics and treatment modalities in the future.
    Matched MeSH terms: Drug Resistance, Bacterial/genetics*
  2. Norazah A, Rasinah WZ, Zaili Z, Aminuddin A, Ramelah M
    Malays J Pathol, 2009 Jun;31(1):29-34.
    PMID: 19694311 MyJurnal
    This study was conducted to determine whether there was any genetic heterogeneity among Helicobacter pylori strains isolated from the antrum and corpus of the same individual in a Malaysian population and to determine the presence of heterogeneous susceptibility of the isolates by comparing PCR-RAPD and antibiotic profiles. Forty-four H. pylori isolates cultured from the antrum and corpus of 22 patients were analyzed. Antibiotic susceptibility testing was carried out by minimum inhibitory concentration determination, using E-Test method strips. PCR-RAPD was carried out on all the strains and the profiles generated were analysed for cluster analysis. Twenty-nine different PCR-RAPD profiles were observed in the 44 isolates. Fifteen pairs of the isolates from the same patients had the same PCR-RAPD patterns while in 7 pairs, the profiles were different. The strains were clustered into 2 separate clusters at a low coefficient of similarity, where most of the strains were in cluster 1. The degree of similarity was very low among most of the isolates. Most of the patients (16 of 22) were infected with strains that have the same antibiotic susceptibility profiles. Out of these, only 10 pairs shared the same PCR-RAPD and antibiotic profiles. Five pairs of isolates with similar PCR-RAPD profiles differed in their antibiotic profiles due to metronidazole resistance in one of the sites. A large degree of genetic heterogeneity was observed among H. pylori strains circulating among Malaysian patients. An individual patient can be infected with multiple strains and the strains can be antibiotic resistant.
    Matched MeSH terms: Drug Resistance, Bacterial/genetics
  3. Learn-Han L, Yoke-Kqueen C, Salleh NA, Sukardi S, Jiun-Horng S, Chai-Hoon K, et al.
    Antonie Van Leeuwenhoek, 2008 Oct;94(3):377-87.
    PMID: 18548329 DOI: 10.1007/s10482-008-9254-y
    Forty-eight strains of Salmonella enterica subsp. enterica serovar Agona and 33 strains of Salmonella enterica subsp. enterica serovar Weltevreden were characterized by random amplified polymorphic DNA (RAPD) fingerprinting using 3 different arbitrary primer, Enterobacterial Repetitive Intergenic Consensus-Polymerase Chain Reaction (ERIC-PCR) and antimicrobial susceptibility testing. By using RAPD, 81 strains (44 strains of S. Agona and 33 strains of S. Weltevreden) can be clustered into 14 groups and 6 single isolates whereas ERIC-PCR produced 7 clusters and 3 single isolates. Thirteen antimicrobial agents were used and all the isolates were resistant to erythromycin and showed Multiple Antimicrobial Resistance indexes, ranging from 0.08 to 0.62. Poultry still remain as the common reservoir for multi-drug-resistant Salmonella. On the other hand, vegetables contaminated with S. Weltevreden showed a gain in antimicrobial resistance. Besides that, consistent antibiograms were observed from S. Weltevreden isolated at Kajang wet market on 2000/08/02.
    Matched MeSH terms: Drug Resistance, Bacterial/genetics
  4. Lye YL, Bong CW, Lee CW, Zhang RJ, Zhang G, Suzuki S, et al.
    Sci Total Environ, 2019 Oct 20;688:1335-1347.
    PMID: 31726563 DOI: 10.1016/j.scitotenv.2019.06.304
    The environmental reservoirs of sulfonamide (SA) resistome are still poorly understood. We investigated the potential sources and reservoir of SA resistance (SR) in Larut River and Sangga Besar River by measuring the SA residues, sulfamethoxazole resistant (SMXr) in bacteria and their resistance genes (SRGs). The SA residues measured ranged from lower than quantification limits (LOQ) to 33.13 ng L-1 with sulfadiazine (SDZ), sulfadimethoxine (SDM) and SMX as most detected. Hospital wastewater effluent was detected with the highest SA residues concentration followed by the slaughterhouse and zoo wastewater effluents. The wastewater effluents also harbored the highest abundance of SMXr-bacteria (107 CFU mL-1) and SRGs (10-1/16S copies mL-1). Pearson correlation showed only positive correlation between the PO4 and SMXr-bacteria. In conclusion, wastewater effluents from the zoo, hospital and slaughterhouse could serve as important sources of SA residues that could lead to the consequent emergence of SMXr-bacteria and SRGs in the river.
    Matched MeSH terms: Drug Resistance, Bacterial/genetics*
  5. Shahimi S, Elias A, Abd Mutalib S, Salami M, Fauzi F, Mohd Zaini NA, et al.
    Environ Sci Pollut Res Int, 2021 Aug;28(32):44002-44013.
    PMID: 33846919 DOI: 10.1007/s11356-021-13665-4
    A total of 24 strains of Vibrio alginolyticus were isolated from cockles (Anadara granosa) and identified for VibA and gyrB genes. All V. alginolyticus isolates were then tested against nine different antibiotics. In this study, the highest percentage of antibiotic resistance was obtained against penicillin (37.50%), followed by ampicillin, vancomycin (12.50%) and erythromycin (8.33%). All of V. alginolyticus isolates were susceptible against streptomycin, kanamycin, tetracycline, chloramphenicol and sulfamethoxazole. Polymerase chain reaction (PCR) assay has confirmed the presence of four antibiotic resistance genes of penicillin (pbp2a), ampicillin (blaOXA), erythromycin (ermB) and vancomycin (vanB). Out of 24 V. alginolyticus isolates, 2 isolates possessed the tdh-related hemolysin (trh) (strains VA15 and VA16) and none for the thermostable direct hemolysin (tdh) gene. Both strains of the tdh-related hemolysin (trh) were susceptible to all antibiotics tested. The multiple antibiotic resistance (MAR) index ranging between 0.2 and 0.3 with 5 antibiograms (A1-A5) was observed. Combination of enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) and antibiotic resistance indicated 18 genome types which showed genetic heterogeneity of those V. alginolyticus isolates. The results demonstrated the presence of V. alginolyticus strain found in cockles can be a potential risk to consumers and can contribute to the deterioration of human health in the study area. Thus, it is essential for local authority to provide the preventive measures in ensuring the cockles are safe for consumption.
    Matched MeSH terms: Drug Resistance, Bacterial/genetics
  6. Abatcha MG, Effarizah ME, Rusul G
    Int J Food Microbiol, 2019 Feb 02;290:180-183.
    PMID: 30342248 DOI: 10.1016/j.ijfoodmicro.2018.09.021
    Salmonella enterica serovar Paratyphi B (S. Paratyphi B) is a major foodborne pathogen distributed all over the world. However, little is known about the antibiotic resistance, genetic relatedness and virulence profile of S. Paratyphi B isolated from leafy vegetables and the processing environment in Malaysia. In this study, 6 S. Paratyphi B isolates were recovered from different vegetables and drain water of processing areas obtained from fresh food markets in Malaysia. The isolates were characterized by antibiogram, Pulsed-field gel electrophoresis (PFGE) and virulence genes. Antibiotic susceptibility test showed that 3 of the isolates were resistant to the antibiotics. These include S. Paratyphi B SP251 isolate, which was resistant to chloramphenicol, ampicillin, sulfonamides and streptomycin; Isolate SP246 which was resistant to chloramphenicol, sulfonamides and streptomycin and Isolate SP235 showing resistance to nalidixic acid only. PFGE subtyped the 6 S. Paratyphi B isolates into 6 distinct XbaI-pulsotypes, with a wide range of genetic similarity (0.55 to 0.9). The isolates from different sources and fresh food markets location were genetically diverse. Thirteen (tolC, orgA, spaN, prgH, sipB, invA, pefA, sofB, msgA, cdtB, pagC, spiA and spvB) out of the 17 virulence genes tested were found in all of the S. Paratyphi B isolates. Another gene (lpfC), was found only in one isolate (SP051). None of the isolates possessed sifA, sitC and ironN genes. In summary, this study provides unique information on antibiotic resistance, genetic relatedness, and virulotyping of S. Paratyphi B isolated from leafy vegetables and processing environment.
    Matched MeSH terms: Drug Resistance, Bacterial/genetics
  7. Hoe CH, Yasin RM, Koh YT, Thong KL
    J Appl Microbiol, 2005;99(1):133-40.
    PMID: 15960673
    Antimicrobial resistance of Shigella sonnei from Malaysia was determined and subtyping was carried by pulsed-field gel electrophoresis (PFGE) to assess the extent of genetic diversity of these strains.
    Matched MeSH terms: Drug Resistance, Bacterial/genetics*
  8. Ahmad N, Zakaria WR, Abdullah SA, Mohamed R
    World J Gastroenterol, 2009 Jul 07;15(25):3161-5.
    PMID: 19575497
    AIM: To characterize the types of mutations present in the 23S rRNA genes of Malaysian isolates of clarithromycin-resistant Helicobacter pylori (H pylori).

    METHODS: Clarithromycin susceptibility of H pylori isolates was determined by E test. Analyses for point mutations in the domain V of 23S rRNA genes in clarithromycin-resistant and -sensitive strains were performed by sequence analysis of amplified polymerase chain reaction products. Restriction fragment length polymorphism was performed using BsaI and MboII enzymes to detect restriction sites that correspond to the mutations in the clarithromycin-resistant strains.

    RESULTS: Of 187 isolates from 120 patients, four were resistant to clarithromycin, while 183 were sensitive. The MIC of the resistant strains ranged from 1.5 to 24 microg/mL. Two isolates had an A2142G mutation and another two had A2143G mutations. A T2182C mutation was detected in two out of four clarithromycin-resistant isolates and in 13 of 14 clarithromycin-sensitive isolates. Restriction enzyme analyses with BsaI and MboII were able to detect the mutations.

    CONCLUSION: Clarithromycin resistance is an uncommon occurrence among Malaysian isolates of H pylori strains and the mutations A2142G and A2143G detected were associated with low-level resistance.

    Matched MeSH terms: Drug Resistance, Bacterial/genetics*
  9. Ho SL, Tan EL, Sam CK, Goh KL
    J Dig Dis, 2010 Apr;11(2):101-5.
    PMID: 20402836 DOI: 10.1111/j.1751-2980.2010.00423.x
    To determine the prevalence of primary clarithromycin resistance amongst Helicobacter pylori (H. pylori) strains in Malaysian patients with gastroduodenal diseases, by using restriction fragment length polymorphism (RFLP) in domain V of 23S rRNA.
    Matched MeSH terms: Drug Resistance, Bacterial/genetics*
  10. Jindal HM, Ramanathan B, Le CF, Gudimella R, Razali R, Manikam R, et al.
    J Biomed Sci, 2018 Feb 15;25(1):15.
    PMID: 29448938 DOI: 10.1186/s12929-018-0414-8
    BACKGROUND: Streptococcus pneumoniae or pneumococcus is a leading cause of morbidity and mortality worldwide, specifically in relation to community-acquired pneumonia. Due to the overuse of antibiotics, S. pneumoniae has developed a high degree of resistance to a wide range of antibacterial drugs.

    METHODS: In this study, whole genome sequencing (WGS) was performed for 10 clinical strains of S. pneumoniae with different levels of sensitivity to standard antibiotics. The main objective was to investigate genetic changes associated with antibiotic resistance in S. pneumoniae.

    RESULTS: Our results showed that resistant isolates contain a higher number of non-synonymous single nucleotide polymorphisms (SNPs) as compared to susceptible isolates. We were able to identify SNPs that alter a single amino acid in many genes involved in virulence and capsular polysaccharide synthesis. In addition, 90 SNPs were only presented in the resistant isolates, and 31 SNPs were unique and had not been previously reported, suggesting that these unique SNPs could play a key role in altering the level of resistance to different antibiotics.

    CONCLUSION: Whole genome sequencing is a powerful tool for comparing the full genome of multiple isolates, especially those closely related, and for analysing the variations found within antibiotic resistance genes that lead to differences in antibiotic sensitivity. We were able to identify specific mutations within virulence genes related to resistant isolates. These findings could provide insights into understanding the role of single nucleotide mutants in conferring drug resistance.

    Study site: University Malaya Medical Centre (UMMC)
    Matched MeSH terms: Drug Resistance, Bacterial/genetics*
  11. Chin KL, Sarmiento ME, Norazmi MN, Acosta A
    Tuberculosis (Edinb), 2018 12;113:139-152.
    PMID: 30514496 DOI: 10.1016/j.tube.2018.09.008
    Tuberculosis (TB), caused by Mycobacterium tuberculosis complex (MTBC), is an infectious disease with more than 10.4 million cases and 1.7 million deaths reported worldwide in 2016. The classical methods for detection and differentiation of mycobacteria are: acid-fast microscopy (Ziehl-Neelsen staining), culture, and biochemical methods. However, the microbial phenotypic characterization is time-consuming and laborious. Thus, fast, easy, and sensitive nucleic acid amplification tests (NAATs) have been developed based on specific DNA markers, which are commercially available for TB diagnosis. Despite these developments, the disease remains uncontrollable. The identification and differentiation among MTBC members with the use of NAATs remains challenging due, among other factors, to the high degree of homology within the members and mutations, which hinders the identification of specific target sequences in the genome with potential impact in the diagnosis and treatment outcomes. In silico methods provide predictive identification of many new target genes/fragments/regions that can specifically be used to identify species/strains, which have not been fully explored. This review focused on DNA markers useful for MTBC detection, species identification and antibiotic resistance determination. The use of DNA targets with new technological approaches will help to develop NAATs applicable to all levels of the health system, mainly in low resource areas, which urgently need customized methods to their specific conditions.
    Matched MeSH terms: Drug Resistance, Bacterial/genetics
  12. Klayut W, Rudeeaneksin J, Srisungngam S, Bunchoo S, Bhakdeenuan P, Phetsuksiri B, et al.
    Trop Biomed, 2022 Dec 01;39(4):483-488.
    PMID: 36602205 DOI: 10.47665/tb.39.4.001
    Tuberculosis (TB) continues to be a major public health problem in Thailand and many countries. Endemic TB and outbreaks of TB drug resistance in the borderlands are particularly important. The Thailand-Myanmar border has extensive cross-border travel that may accelerate TB's spread. This cross-sectional study aimed to determine the frequency and factors associated with TB, and rifampicinresistant TB (RR-TB) among presumptive tuberculosis patients in Mae Sot Hospital. Sputum was processed by microscopic examination and Xpert MTB/RIF assay. Laboratory results and socio-demographic characteristics were collected and analyzed. Univariate and multivariate analyses were performed to assess the association of the risk factors with TB and RR-TB. The significant variables at p-values < 0.05 in univariate analysis were selected for multivariate analysis. Of 365 presumptive patients enrolled, 244 (66.85%) were males and 199 (54.52%) were Burmese. Of these, 314 (86.03%) were registered as new cases and 183 (50.14%) worked as laborers. Sputum microscopy was positive in 132 (36.16%) cases. Based on Xpert MTB/RIF, the frequency of TB was 136 (37.26%) and RR-TB was 15 (11.03%). TB was more common in males than females. The majority of the cases belonged to the 26-50-year-old age group and migrant workers. In RR-TB detection, the rpoB mutations covered by probe E were the most frequently observed. Sequencing showed that the most highly mutated codon was codon 531 and Ser531Thr was the most common mutation. For risk factor analysis, working as laborers was significantly (p-value < 0.05) associated with TB (aOR 2.83; 95% CI 1.43-5.63) and previously treated cases were significantly associated with RR-TB (aOR 12.33; 95% CI 2.29-66.49). The high frequency of TB and RR-TB in migrants highlights the problem and factors associated with TB at the border and the need for efforts in TB control programs in this setting.
    Matched MeSH terms: Drug Resistance, Bacterial/genetics
  13. 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: Drug Resistance, Bacterial/genetics*
  14. Ara B, Urmi UL, Haque TA, Nahar S, Rumnaz A, Ali T, et al.
    Expert Rev Clin Pharmacol, 2021 Apr;14(4):513-522.
    PMID: 33691556 DOI: 10.1080/17512433.2021.1901577
    Background: Currently, colistin-resistant pathogens emerged has become a global health concern. This study assessed the distribution of mcr-1 to mcr-5 variants with the phenotypic colistin-resistance in bacterial isolates from urinary tract infection (UTI) patients in Bangladesh.Methods: A cross-sectional study was conducted between April 2017 and March 2018 to enroll uncomplicated UTI patients, and 142 urine samples were analyzed. Uropathogens were identified using the API-20E biochemical panel and 16s rRNA gene sequencing. Polymerase chain reactions detected the mcr gene variants in the UTI isolates. The phenotypic colistin-susceptibility was determined by the Kirby-Bauer disc-diffusion method and the minimal inhibitory concentration (MIC) measurement.Results: The combined carriage of mcr-1 and mcr-2 genes in 11.4% (14/123) of urinary tract pathogens. The mcr-positive pathogens include five Escherichia coli, three Klebsiella pneumoniae, three Pseudomonas putida, two Enterobacter cloacae, and one Enterobacter hormaechei. The mcr-positive variant showed significantly higher phenotypic colistin resistance with MIC between >16 µg/mL and >128 µg/mL (pdrug may lead to a lack of treatment options for the infectious diseases and spread of infection to the unaffected cohorts.
    Matched MeSH terms: Drug Resistance, Bacterial/genetics
  15. Yang SK, Yusoff K, Ajat M, Thomas W, Abushelaibi A, Akseer R, et al.
    PLoS One, 2019;14(4):e0214326.
    PMID: 30939149 DOI: 10.1371/journal.pone.0214326
    Klebsiella pneumoniae (KP) remains the most prevalent nosocomial pathogen and carries the carbapenemase (KPC) gene which confers resistance towards carbapenem. Thus, it is necessary to discover novel antimicrobials to address the issue of antimicrobial resistance in such pathogens. Natural products such as essential oils are a promising source due to their complex composition. Essential oils have been shown to be effective against pathogens, but the overall mechanisms have yet to be fully explained. Understanding the molecular mechanisms of essential oil towards KPC-KP cells would provide a deeper understanding of their potential use in clinical settings. Therefore, we aimed to investigate the mode of action of essential oil against KPC-KP cells from a proteomic perspective by comparing the overall proteome profile of KPC-KP cells treated with cinnamon bark (Cinnamomum verum J. Presl) essential oil (CBO) at their sub-inhibitory concentration of 0.08% (v/v). A total of 384 proteins were successfully identified from the non-treated cells, whereas only 242 proteins were identified from the CBO-treated cells. Proteins were then categorized based on their biological processes, cellular components and molecular function prior to pathway analysis. Pathway analysis showed that CBO induced oxidative stress in the KPC-KP cells as indicated by the abundance of oxidative stress regulator proteins such as glycyl radical cofactor, catalase peroxidase and DNA mismatch repair protein. Oxidative stress is likely to oxidize and disrupt the bacterial membrane as shown by the loss of major membrane proteins. Several genes selected for qRT-PCR analysis validated the proteomic profile and were congruent with the proteomic abundance profiles. In conclusion, KPC-KP cells exposed to CBO undergo oxidative stress that eventually disrupts the bacterial membrane possibly via interaction with the phospholipid bilayer. Interestingly, several pathways involved in the bacterial membrane repair system were also affected by oxidative stress, contributing to the loss of cells viability.
    Matched MeSH terms: Drug Resistance, Bacterial/genetics
  16. Kim SY, Ko KS
    Microb Drug Resist, 2019 Mar;25(2):227-232.
    PMID: 30212274 DOI: 10.1089/mdr.2018.0020
    To reveal whether an increase of CTX-M-15-producing Klebsiella pneumoniae ST11 isolates is due to clonal dissemination across the countries, plasmids (pHK02-026, pM16-13, pIN03-01, and pTH02-34) were extracted from four K. pneumoniae isolates collected in Hong Kong, Malaysia, Thailand, and Indonesia, respectively. Complete sequencing of blaCTX-M-15-carrying plasmids was performed. In addition to the four plasmids, a previously sequenced plasmid (pKP12226) of a K. pneumoniae ST11 isolate from Korea was included in the analysis. While pIN03-01 and pTH02-34, which belonged to the incompatibility group IncX3, showed nearly the same structure, the others of IncF1A or IncFII exhibited very different structures. The number and kinds of antibiotic genes found in the plasmids were also different from each other. Cryptic prophage genes were identified in all five blaCTX-M-15-harboring plasmids from the ST11 isolates; P1-like region in pKP12226, CPZ-55 prophage region in pHK02-026, phage shock operon pspFABCD in pM16-13, and SPBc2 prophage yokD in pIN03-01 and pTH02-34. The plasmids with blaCTX-M-15 in the prevailing K. pneumoniae ST11 isolates in Asian countries might emerge from diverse origins by recombination. The prevalence of CTX-M-15-producing K. pneumoniae ST11 clone in Asian countries is not mainly due to the dissemination of a single strain.
    Matched MeSH terms: Drug Resistance, Bacterial/genetics*
  17. Rameshkumar MR, Arunagirinathan N, Swathirajan CR, Vignesh R, Balakrishnan P, Solomon SS
    Indian J Med Res, 2018 09;148(3):341-344.
    PMID: 30425226 DOI: 10.4103/ijmr.IJMR_730_17
    Matched MeSH terms: Drug Resistance, Bacterial/genetics
  18. Wahab HA, Choong YS, Ibrahim P, Sadikun A, Scior T
    J Chem Inf Model, 2009 Jan;49(1):97-107.
    PMID: 19067649 DOI: 10.1021/ci8001342
    The continuing rise in tuberculosis incidence and the problem of drug resistance strains have prompted the research on new drug candidates and the mechanism of drug resistance. Molecular docking and molecular dynamics simulation (MD) were performed to study the binding of isoniazid onto the active site of Mycobacterium tuberculosis enoyl-acyl carrier protein reductase (InhA) in an attempt to address the mycobacterial resistance against isoniazid. Results show that isonicotinic acyl-NADH (INADH) has an extremely high binding affinity toward the wild type InhA by forming stronger interactions compared to the parent drug (isoniazid) (INH). Due to the increase of hydrophobicity and reduction in the side chain's volume of A94 of mutant type InhA, both INADH and the mutated protein become more mobile. Due to this reason, the molecular interactions of INADH with mutant type are weaker than that observed with the wild type. However, the reduced interaction caused by the fluctuation of INADH and the mutant protein only inflected minor resistance in the mutant strain as inferred from free energy calculation. MD results also showed there exists a water-mediated hydrogen bond between INADH and InhA. However, the bridged water molecule is only present in the INADH-wild type complex, reflecting the putative role of the water molecule in the binding of INADH to the wild type protein. The results support the assumption that the conversion of prodrug isoniazid into its active form INADH is mediated by KatG as a necessary step prior to target binding on InhA. Our findings also contribute to a better understanding of INH resistance in mutant type.
    Matched MeSH terms: Drug Resistance, Bacterial/genetics
  19. Low YM, Chong CW, Yap IKS, Chai LC, Clarke SC, Ponnampalavanar S, et al.
    Pathog Glob Health, 2018 10;112(7):378-386.
    PMID: 30380366 DOI: 10.1080/20477724.2018.1538281
    The increasing prevalence of antibiotic resistant pathogens poses a serious threat to global health. However, less emphasis has been placed to co-relate the gene expression and metabolism of antibiotic resistant pathogens. This study aims to elucidate gene expression and variations in metabolism of multidrug resistant Klebsiella pneumoniae after exposure to antibiotics. Phenotypic responses of three genotypically distinct carbapenem resistant Klebsiella pneumoniae (CRKP) strains untreated and treated with sub-lethal concentrations of imipenem were investigated via phenotype microarrays (PM). The gene expression and metabolism of the strain harboring blaNDM-1 before and after exposure to sub-lethal concentration of imipenem were further investigated by RNA-sequencing (RNA-Seq) and 1H NMR spectroscopy respectively. Most genes related to cell division, central carbon metabolism and nucleotide metabolism were downregulated after imipenem treatment. Similarly, 1H NMR spectra obtained from treated CRKP showed decrease in levels of bacterial end products (acetate, pyruvate, succinate, formate) and metabolites involved in nucleotide metabolism (uracil, xanthine, hypoxanthine) but elevated levels of glycerophosphocholine. The presence of anserine was also observed for the treated CRKP while FAPγ-adenine and methyladenine were only present in untreated bacterial cells. As a conclusion, the studied CRKP strain exhibited decrease in central carbon metabolism, cell division and nucleotide metabolism after exposure to sub-lethal concentrations of imipenem. The understanding of the complex biological system of this multidrug resistant bacterium may help in the development of novel strategies and potential targets for the management of the infections.
    Matched MeSH terms: Drug Resistance, Bacterial/genetics
  20. Veldman K, Kant A, Dierikx C, van Essen-Zandbergen A, Wit B, Mevius D
    Int J Food Microbiol, 2014 May 2;177:72-7.
    PMID: 24607424 DOI: 10.1016/j.ijfoodmicro.2014.02.014
    Since multidrug resistant bacteria are frequently reported from Southeast Asia, our study focused on the occurrence of ESBL-producing Enterobacteriaceae in fresh imported herbs from Thailand, Vietnam and Malaysia. Samples were collected from fresh culinary herbs imported from Southeast Asia in which ESBL-suspected isolates were obtained by selective culturing. Analysis included identification by MALDI-TOF mass spectrometry, susceptibility testing, XbaI-PFGE, microarray, PCR and sequencing of specific ESBL genes, PCR based replicon typing (PBRT) of plasmids and Southern blot hybridization. In addition, the quinolone resistance genotype was characterized by screening for plasmid mediated quinolone resistance (PMQR) genes and mutations in the quinolone resistance determining region (QRDR) of gyrA and parC. The study encompassed fifty samples of ten batches of culinary herbs (5 samples per batch) comprising nine different herb variants. The herbs originated from Thailand (Water morning glory, Acacia and Betel leaf), Vietnam (Parsley, Asian pennywort, Houttuynia leaf and Mint) and Malaysia (Holy basil and Parsley). By selective culturing 21 cefotaxime resistant Enterobacteriaceae were retrieved. Array analysis revealed 18 isolates with ESBL genes and one isolate with solely non-ESBL beta-lactamase genes. Mutations in the ampC promoter region were determined in two isolates with PCR and sequencing. The isolates were identified as Klebsiella pneumoniae (n=9), Escherichia coli (n=6), Enterobacter cloacae complex (n=5) and Enterobacter spp. (n=1). All isolates tested were multidrug resistant. Variants of CTX-M enzymes were predominantly found followed by SHV enzymes. PMQR genes (including aac(6')-1b-cr, qnrB and qnrS) were also frequently detected. In almost all cases ESBL and quinolone resistance genes were located on the same plasmid. Imported fresh culinary herbs from Southeast Asia are a potential source for contamination of food with multidrug resistant bacteria. Because these herbs are consumed without appropriate heating, transfer to human bacteria cannot be excluded.
    Matched MeSH terms: Drug Resistance, Bacterial/genetics*
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