Ming-Li Liou

Clonal spread of multidrug-resistant Acinetobacter baumannii in eastern Taiwan

BACKGROUND AND PURPOSE This study was conducted to investigate the molecular epidemiology and antimicrobial susceptibility of multidrug-resistant (MDR) Acinetobacter baumannii to three types of antibiotics. METHODS One hundred and thirty-four specimens of MDR A baumannii were collected from three branches (Taipei, Dalin, and Hualien branches) of Buddhist Tzu Chi Hospital, which are located in northern, southern, and eastern Taiwan, during 2007. Genotyping was performed by pulsed-field gel electrophoresis. Antibiotic susceptibilities to colistin, rifampicin, and tigecycline were determined. The synergistic effects of rifampin and colistin were also evaluated. RESULTS Antibiotic susceptibility testing showed that 10.4%, 47.8% and 45.5% of the MDR A baumannii isolates are resistant to colistin, rifampicin, and tigecycline, respectively. A majority of the rifampicin-resistant isolates (62.7%) were found in the Haulien branch, whereas 62.2% of tigecycline-resistant isolates were found in the Taipei branch. The combination of colistin and rifampicin had a synergistic effect on all of the isolates. Genotyping by pulsed-field gel electrophoresis identified 17, 23, and 11 pulsotypes in the Taipei, Dalin, and Haulien branches, respectively. Furthermore, 74.5% of isolates in the Haulien branch were identified as one of three pulsotypes. Among 37 rifampicin-resistant and 22 tigecycline-resistant MDR A baumannii isolates found in the Haulien branch, 51.3% (19/37) and 50% (11/22) of the isolates belonged to the same clone, respectively. CONCLUSION This study confirms the high prevalence of resistance to rifampicin and tigecycline in MDR A baumannii in the three hospitals that were studied, and the high proportion of identical strains that exist in eastern Taiwan.

Imipenem: a potent inducer of multidrug resistance in Acinetobacter baumannii

This study investigated the progression of multidrug resistance upon exposure to imipenem in Acinetobacter baumannii. Eighteen A. baumannii strains, including two reference strains (ATCC 19606 and ATCC 17978), four clinical strains (AB56, AB242, AB273 and AB279) and 12 antibiotic-selected mutant strains, were used in this study. Imipenem-selected mutants were generated from imipenem-susceptible strains (ATCC 19606, ATCC 17978 and AB242) by multistep selection resistance. Amikacin-, ciprofloxacin-, colistin-, meropenem- and ceftazidime-selected mutants were also generated from the two reference strains and were used for comparison. Antibiotic susceptibilities in the absence and presence of the efflux pump inhibitors carbonyl cyanide m-chlorophenylhydrazone (CCCP) and 1-(1-naphthylmethyl)-piperazine (NMP) were examined in the three imipenem-selected mutants and the three clinical multidrug-resistant (MDR) isolates. Expression profiles of the antimicrobial resistance genes in the imipenem-selected mutants and their parental strains were also determined. The results showed that imipenem was more likely, compared with the other antibiotics, to induce a MDR phenotype in the two reference strains. Differences in OXA-51-like carbapenemase, efflux pumps or/and AmpC β-lactamase expression were observed in the three imipenem-selected mutants. Moreover, a reduction in imipenem or amikacin resistance was observed when the imipenem-selected mutants and clinical isolates were exposed to NMP and CCCP. This study concluded that imipenem might be a potent inducer of multidrug resistance in A. baumannii strains. OXA-51-like carbapenemase combined with other resistance mechanisms may contribute to the development of multidrug resistance in A. baumannii. Monitoring the use of carbapenems is required to reduce the spread of MDR A. baumannii in hospitals.

Comparison of antimicrobial resistance patterns between clinical and sewage isolates in a regional hospital in Taiwan

Aims:  To compare bacterial populations and antimicrobial resistance patterns between clinical and sewage isolates from a regional hospital in northern Taiwan. The dissemination of antibiotic‐resistant bacteria from hospital compartments to the hospital sewage treatment plant was examined.

The sensor kinase BfmS mediates virulence in Acinetobacter baumannii

BACKGROUND/PURPOSE BfmR, the response regulator component of the two-component system BfmRS, has important roles in biofilm formation and cellular morphology of Acinetobacter baumannii. Until now, the contribution of the sensor kinase BfmS to the virulence of this bacterium remains unknown. In this study, a bfmS knockout and complementation studies were performed to clarify the role of BfmS in A. baumannii virulence. METHODS We constructed a bfmS knockout mutant in the A. baumannii 17978 type strain by transposon inactivation. To clarify the role of bfmS in A. baumannii virulence, the biofilm formation, adherence ability to eukaryotic cells, serum resistance, and antibiotic susceptibility tests were performed in A. baumannii 17978 and its derivative knockout and complementation strains. RESULTS The bfmS knockout displayed a reduction in biofilm formation, loss of adherence to eukaryotic cells, and greater sensitivity to serum killing compared with the parent strain. Proteomic analysis of culture supernatants revealed that the release of outer membrane proteins (Omps), including CarO and outer membrane protein A (OmpA), was associated with the inactivation of BfmS in A. baumannii. CONCLUSION This study is the first to demonstrate that the pathway regulated by the sensor kinase BfmS is associated with biofilm formation, adherence to biotic surfaces, serum resistance, and antibiotic susceptibility, which may be associated with the release of Omps in A. baumannii.

Distribution of blaOXA-carrying imipenem-resistant Acinetobacter spp. in 3 hospitals in Taiwan.

We investigated the molecular epidemiology and OXA-type carbapenemase genes of 83 imipenem-resistant Acinetobacter spp. collected from 2 university hospitals (hospitals A and B) and a regional hospital (hospital C) during 2007 in Taiwan. Genotyping by pulsed-field gel electrophoresis identified 51 pulsotypes. None of the pulsotypes established predominance throughout the 3 hospitals. Multiplex polymerase chain reaction of blaOXA genes showed that 100% (18/18), 91%(31/34), and 100% (31/31) of the Acinetobacter spp. collected from hospital A, B, and C, respectively, possessed blaOXA-51-like genes. None of the strains carrying blaOXA-23-like and blaOXA-24-like genes were found in hospital A. The coexistences of blaOXA-51-like/blaOXA-23-like and blaOXA-51-like/blaOXA-24-like genes detected in hospitals B and C were 26% (9/34) and 12% (4/34) and 58% (18/31) and 3% (1/31), respectively. Among blaOXA-23-like gene-carrying isolates collected from hospitals, clonal spread of strains carrying the blaOXA-23 gene was detected in the regional hospital but not the other 2 university hospitals. The results suggest that interhospital dissemination of imipenem-resistant Acinetobacter spp. was not found in these hospitals. The increasing percentage of OXA-23 in OXA-type carbapenemases in Acinetobacter spp. from the regional hospitals to medical centers deserves further attention in Taiwan.

The origin of Acinetobacter baumannii TYTH-1: a comparative genomics study

There have been increasing reports of bla(OXA-23)-carrying strains of carbapenem-resistant Acinetobacter baumannii (CRAB), which has become a significant public health concern in Taiwan. To determine the origin of these CRAB strains, the prevalence of CRAB and bla(OXA-23)-carrying CRAB in a regional hospital was analysed retrospectively. The genome of A. baumannii TYTH-1 was completely sequenced and annotated. Multiple comparative genomics studies, including phylogenetic analysis, functional comparison via the Clusters of Orthologous Groups (COGs) database, and determination of variance in GC profiles in the whole genome and gene arrangements in resistance islands, were performed using 11 completely sequenced A. baumannii genomes. bla(OXA-23)-carrying CRAB isolates became dominant clones in 2007. A comparative genomics analysis revealed a common strain lineage between Taiwanese strains (TYTH-1 and TCDC-AB0715) and Chinese strains (MDR-TJ and MDR-ZJ06). Phylogenetic studies and GC profiles showed that the genome of TYTH-1 was closest to MDR-ZJ06. However, the resistance island of TYTH-1 (RI(TYTH-1)) was nearly identical to that of RI(MDT-TJ). The functional category for COGs was similar in the tested genomes. The results reveal that dissemination of bla(OXA-23)-carrying CRAB in Taiwan may have been mediated by the transfer of people between Taiwan and China during 2007. The global spread of CRAB is now a worldwide public health problem. In Taiwan, the government needs to focus more attention on the importance of identifying and tracing resistant pathogens and issuing notifications of CRAB infections.

Emergence and dissemination of blaOXA-23-carrying imipenem-resistant Acinetobacter sp in a regional hospital in Taiwan

BACKGROUND The distribution and characterization of OXA-type carbapenemases in Acinetobacter sp in Taiwan has less been reported. The aim of the study was to investigate the molecular epidemiology and OXA-type carbapenemase genes in a regional hospital in Taiwan. METHODS Imipenem-resistant Acinetobacter sp were collected between 2005 and 2007 in a regional hospital. Genotyping was performed by pulsed-field gel electrophoresis. OXA-type carbapenemase genes were determined by multiplex polymerase chain reaction (PCR) and gene sequencing. RESULTS A total of 136 isolates were collected. Fifty-six pulsotypes were identified. None of the pulsotypes established predominance throughout the 3-year period. Multiplex PCR of blaOXA genes showed that 99% (135/136) of the Acinetobacter sp possessed blaOXA51-like genes. The coexistences of blaOXA51-like/blaOXA-23-like and blaOXA51-like/blaOXA-24-like were detected in 19% (26/136) and 1% (2/136) of the isolates, respectively. Among blaOXA-23-like gene-carrying isolates, two isolates (Pulsotypes 18 and 20) were found in 2006 and the remainder (n=24), including Pulsotypes 27 (n=18), 29 (n=1), 52 (n=3), and 53 (n=2), were found in 2007. Sequencing performed on the 26 representative isolates confirmed the presence of the blaOXA-23 carbapenemase gene. Analysis of the genetic content of blaOXA-23 showed that these genes were presumably chromosomal and associated with the upstream-located insertion sequence ISAba1. CONCLUSIONS The emergence and imminent widespread of blaOXA-23-carrying imipenem-resistant Acinetobacter sp appeared in Taiwan during the period from 2006 to 2007.

Genome Sequence of Acinetobacter baumannii TYTH-1

Acinetobacter baumannii has emerged recently as a major cause of health care-associated infections due to the extent of its antimicrobial resistance and its propensity to cause large nosocomial outbreaks. Here we report the genome sequence of Acinetobacter baumannii TYTH-1 isolated in Taiwan during 2008.

Clinical features and molecular epidemiology of multidrug-resistant Acinetobacter calcoaceticus-A baumannii complex in a regional teaching hospital in Taiwan

We conducted a case-controlled study in a regional teaching hospital in Taiwan to investigate the clinical features and molecular epidemiology of multidrug-resistant Acinetobacter calcoaceticus-A baumannii (MDR Acb) complex. Case patients had higher mortality than controls did. MDR Acb complex acquisition risk factors include longer hospital stays, higher ratio of nasogastric tube and Foley catheter use, and more carbapenem use. All available isolates were divided into 36 subtypes by pulsed-field gel electrophoresis. The proportion of the same subtypes with their appearance within 1 and 2 months was 62.5% and 87.5%, respectively. We concluded that many different MDR Acb complex clones could be found in a hospital and that the same clones often spread on a small scale within a short period of time if no outbreaks noted.

Subgingival microbiota in individuals with severe chronic periodontitis

BACKGROUND/PURPOSE Subgingival microorganisms are potentially associated with periodontal diseases. However, the correlation between the variance in the periodontal microbiome and the prevalence and severity of periodontitis remains unclear. The aim of this study was to determine the subgingival microbiota in Taiwanese individuals with severe chronic periodontitis (SP). METHODS The composition of the subgingival microbiota in healthy and diseased individuals was compared using a 16S rRNA metagenomic approach and quantitative polymerase chain reaction (qPCR). A total of 20 samples, including 10 from healthy individuals and 10 from SP patients, were analyzed. RESULTS We found high microbial diversity, with an average of 774 classified phylotypes per sample and a total of six bacterial phyla across all samples. Cluster analysis by principal component analysis and heat map showed that the bacterial communities were different in the two groups. Streptococcus dominated across all the healthy samples, whereas Prevotella, Porphyromonas, and Treponema were highly abundant across all diseased samples. At least 13 bacterial genera were conserved among all the samples. Only eight genera, including Lautropia, Parvimonas, Actinomyces, Capnocytophaga, Paludibacter, Streptococcus, Haemophilus, and Corynebacterium, were significantly enriched in the healthy group, and six genera, including Porphyromonas, Treponema, Tannerella, Aggregatibacter, Peptostreptococcus, and Filifactor, were significantly enriched in the diseased group. Furthermore, a trend of abundance of bacteria at the species level measured by qPCR in all samples was consistent with the 16S rRNA metagenomics results. CONCLUSION This study is the first in Taiwan to provide a picture of the microbiome in SP via 16S rRNA metagenomics.