Aims: The surplus use of herbicide Dalapon® contains 2,2-dichloropropionic acid (2,2-DCP) poses great danger to human and ecosystem due to its toxicity. Hence, this study focused on the isolation and characterization of a dehalogenase producing bacteria from Sungai Skudai, Johor, capable of utilizing 2,2-DCP as a carbon source and in silico analysis of its putative dehalogenase. Methodology and results: Isolation of the target bacteria was done by using 2,2-DCP-enriched culture as the sole carbon source that allows a bacterium to grow in 20 mM of 2,2-DCP at 30 °C with the corresponding doubling time of 8.89 ± 0.03 h. The isolated bacterium was then designated as Klebsiella pneumoniae strain YZ based on biochemical tests and basic morphological examination. The full genome of K. pneumoniae strain KLPN_25 (accession number: RRE04903) which obtained from NCBI database was screened for the presence of dehalogenase gene, assuming both strains YZ and KLPN_25 were the same organisms. A putative dehalogenase gene was then identified as type II dehalogenase from the genome sequence of strain KLPN_25. The protein structure of the type II dehalogenase of KLPN_25 strain was then pairwise aligned with the crystal structure of L-2-haloacid dehalogenase (L-DEX) Pseudomonas sp. strain YL as the template, revealing the existence of conserved amino acids residues, uniquely known to participate in the dehalogenation mechanism. The finding thus implies that the amino acid residues of type II dehalogenase possibly shares similar catalytic functions with the L-DEX. Conclusion, significance and impact of the study In conclusion, this study confirmed the presence of new dehalogenase from the genus Klebsiella with potential to degrade 2,2-DCP from the river water. The structural information of type II dehalogenase provides insights for future work in designing haloacid dehalogenases.
Klebsiella pneumoniae--isolation & ; purification ; Computer Simulation ; Molecular Dynamics Simulation

INTRODUCTION: In recent years, () has emerged as the predominant cause of pyogenic liver abscess in Asia. - as the causative microorganism in other visceral organ abscesses-is less described. In this study, we seeked to describe the clinical characteristics of visceral organ abscesses in our institution and evaluated the prescription practices of physicians with regard to antibiotic therapy. MATERIALS AND METHODS: A retrospective analysis of patients with culture positive (blood or abscess aspirate) visceral organ abscesses from May 2014 to April 2016 requiring hospitalisation in Changi General Hospital was conducted. RESULTS: A total of 140 adult patients with visceral organ abscesses were identified. The commonest site of involvement was the liver (77.9%), followed by genitourinary tract (20.7%). Diabetic patients were more likely to have liver abscesses, genitourinary abscesses, abscesses in 2 or more organs, genitourinary disease with abscess formation outside of the genitourinary tract, and endovascular infection. Patients with extended spectrum beta-lactamase producing , were more likely to have an obstructive lesion related to the site of the abscess. Overall mortality rate was 7.1%. Amongst survivors, the mean total duration of parenteral antimicrobial therapy was 2.5 weeks before switching to oral antimicrobial agents. CONCLUSION Genitourinary tract is the commonest extra-hepatic site for visceral organ abscess in infections. Parenteral to oral switch of antimicrobial agents appears to be a safe and effective treatment option.
Abscess ; classification ; microbiology ; mortality ; therapy ; Anti-Bacterial Agents ; therapeutic use ; Diabetes Mellitus ; epidemiology ; Female ; Humans ; Klebsiella pneumoniae ; isolation & purification ; Liver ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Risk Factors ; Singapore ; epidemiology ; Survival Analysis ; Urogenital System ; pathology ; Viscera ; pathology

Objective The aim of our study is to investigate the prevalence of Carbapenem-resistant Klebsiella pneumoniae (CRKP) and the genetic characteristics of the class 1 integron in CRKP on multi-drug resistance.Methods Clinical Klebsiella pneumoniae strains were collected from multiple departments of a hospital in central China. CRKP strains were identified among the isolates, and antibiotics susceptibility of CRKP strains was analyzed. The polymerase chain reaction (PCR) was adopted to amplify the class 1 integron variable area. The integron genetic structure was analyzed with enzyme digestion and DNA sequencing technology. The relation between class 1 integron and drug resistance was analyzed statistically.Results Totally 955 strains of Klebsiella pneumoniae were isolated from varied sites of the hospital, and 117(12.3%) of them were identified as CRKP, with a separation rate of 8.9% (26/292) in 2013, 11.3% (38/336) in 2014 and 16.2% (53/327) in 2015, which shows an increasing trend by year. 44.4% (52/117) of CRKP strains were separated from specimen of ICU, and 61.5% (72/117) were from sputum. Over 95% CRKP strains were resistant to ampicillin/sulbactam, aztreonam, imipenem, meropenem, ceftazidme, cefotaxime, cefepime,and piperacillin, while relatively low resistant rates were found in tigecycline (12.8%) and colistin (35.9%). The class 1 integron was detected in 77.8% (91/117) of CRKP strains. Class 1 integron of CRKP was significantly correlated with the antibiotic resistance to the tobramycin, gentamicin and amikacin (all P<0.01). The gene cassette analysis of variable area of class 1 integron showed that aadA2 accounts for 64.8% (59/91), aacA4-catB8-aadA1 23.1% (21/91), and aadA2-dfrA25 12.1% (11/91).Conclusions CRKP has an increasing trend in a clinical setting in China, and most of them were resistant to multiple antibiotics. Class 1 integron in CRKP has strong ability to capture the genes resistant to aminoglycosides antibiotics from environment, with the aadA2 gene as the most popular one.
Anti-Bacterial Agents ; pharmacology ; Carbapenems ; pharmacology ; Drug Resistance, Bacterial ; Integrons ; Klebsiella pneumoniae ; drug effects ; genetics ; isolation & purification

PURPOSE: Tigecycline is one of the drugs used to treat multi-drug resistant Klebsiella pneumoniae (K. pneumoniae) infections, including complicated skin and soft tissue infections, complicated intra-abdominal infection, and community-acquired pneumonia in the Republic of Korea. However, since its commercial release, K. pneumoniae resistance against tigecycline has been reported, and there is a serious concern about the spread of tigecycline resistant bacteria. MATERIALS AND METHODS: In this study, we collected and analyzed 342 isolates from 23 hospitals in the Republic of Korea to determine the mechanisms of tigecycline susceptibility and their clonal types. The hospitals include several from each province in the Republic of Korea, except Jeju, an island province, and nonsusceptibility among the isolates was tested by the disk diffusion method. In our lab, susceptibility was checked again using the broth dilution method, and clonal types were determined using the multilocus sequence typing protocol. Real-time PCR was performed to measure the ramR mutation in the isolates nonsusceptible to tigecycline, which would suggest an increased expression of the AcrAB multidrug pump. RESULTS: Fifty-six K. pneumoniae isolates were found to be nonsusceptible, 16% of the 342 collected. Twenty-seven and nine isolates of the tigecycline nonsusceptible isolates had mutations in the ramR and rpsJ genes, respectively; while 18 nonsusceptible isolates harbored the tetA gene. Comparison of isolates with and without ramR mutation showed a significant statistical difference (p<0.05) for expression of AcrAB. Moreover, the most common clonal types, as observed in our study, appear to be ST11 and ST789. CONCLUSION: Several dominate clonal types infer tigecycline resistance to K. pneumoniae, including ST11, ST768, ST15, ST23, ST48, and ST307. There does not seem to be a transferrable medium, such as plasmid, for the resistance yet, although mutation of the ramR gene may be a common event, accounting for 48% of the nonsusceptibility in this study.
Anti-Bacterial Agents/*pharmacology/therapeutic use ; Bacterial Proteins ; *Drug Resistance, Bacterial ; Humans ; Klebsiella Infections/*drug therapy ; Klebsiella pneumoniae/*drug effects/genetics/*isolation & purification ; Microbial Sensitivity Tests ; Minocycline/*analogs & derivatives/pharmacology/therapeutic use ; Multilocus Sequence Typing ; Plasmids ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction ; Republic of Korea

Carbapenemase-producing organisms (CPO) are rapidly disseminating worldwide, and their presence in tertiary care hospitals poses a significant threat to the management of nosocomial infections. There is a need to control CPO, especially in intensive care unit (ICU) patients, because these organisms are resistant to most beta-lactam antibiotics and are easily transmitted. At present, the identification of CPO is time-consuming; hence, this study focused on the use of the Xpert CARBA-R assay (Cepheid, USA) to determine intestinal colonization rates of CPO in patients admitted to the ICU of a tertiary care hospital in Korea. Forty clinical stool samples were collected and inoculated both in a CARBA-R cartridge and in conventional culture plates. The CARBA-R assay required only ~one hour to screen CPO, while the time required for conventional culture was over three days. We also found that the prevalences of intestinal colonization by carbapenem-resistant organisms and Enterobacteriaceae were 17.5% (7 out of 40) and 7.5% (3 out of 40), respectively. Among the colonizing strains, three that contained carbapenemase, including Klebsiella pneumonia carbapenemase (KPC), and imipenem (IMP) and Verona integron-mediated metallo-beta-lactamase (VIM) were found. With its convenience, the Xpert CARBA-R assay can be included in CPO surveillance strategies.
Anti-Bacterial Agents/pharmacology ; Bacterial Proteins/*genetics/metabolism ; DNA, Bacterial/analysis ; Drug Resistance, Multiple, Bacterial/genetics ; Enterobacteriaceae/drug effects/genetics/*isolation & purification ; Feces/microbiology ; Humans ; Imipenem/pharmacology ; Intensive Care Units ; Klebsiella pneumoniae/drug effects/genetics/*isolation & purification ; Reagent Kits, Diagnostic ; Real-Time Polymerase Chain Reaction ; Republic of Korea ; Tertiary Healthcare ; beta-Lactamases/*genetics/metabolism

Objective To investigate the genotype of klebsiella pneumonia strains isolated from eldly inpatients by multiple-locus variable-number tandem-repeat analysis. Methods Totally 184 klebsiella pneumonia strains,isolated from eldly inpatients,were collected,and their genome DNA were extracted. The polymorphism of 7 variable-number tandem-repeat locus in the DNA samples was analyzed by multiple primers polymerase chain reaction and capillary electrophoresis. The clustering analysis of genotyping was carried out with the BioNumerics 5.1 software. Results A total of 139 genotypes were identified in 184 klebsiella pneumonia clinical strains,showing obvious genetic polymorphisms. With clustering analysis of genotypes,all the strains were categorized into three gene clusters (genogroups 1,2,and 3). The genogroup 1 was the biggest cluster,containing 93.06% of the isolated strains. Conclusion There was a predominant cluster in the klebsiella pneumonia strains isolated from eldly inpatients in our center,and the major source of klebsiella pneumonia infection remained the nosocomial infection.
Aged ; Bacterial Typing Techniques ; Cross Infection ; Genotype ; Genotyping Techniques ; Humans ; Inpatients ; Klebsiella pneumoniae ; classification ; isolation & purification ; Minisatellite Repeats ; Polymerase Chain Reaction ; Polymorphism, Genetic

OBJECTIVETo compare the characteristics of urinary tract infection (UTI) between kidney transplant recipients and non-recipient patients.

METHODSForty-nine kidney transplant recipients with UTI (69 episodes) and 401 non-recipient patients with UTI (443 episodes) admitted in Nanfang Hospital from January 2003 to August 2014 were enrolled in the study. The characteristics of UTI were compared between two groups.

RESULTSIn both groups of UTI, female patients comprised a greater proportion (63.3% and 58.6%) and Escherichia coli was the most common pathogen isolated (37.7% and 34.1%). However, the infection rate of Klebsiella pneumonia in recipients was higher than that in non-recipients (11.6% vs 3.2%, P= 0.001), while the infection rate of Candida albicans was lower (1.5% vs 11.3%, P=0.008) than that in non-recipients. Recipients were likely to develop antibiotic resistance and with a higher recurrence rate than non-recipient patients (38.8% vs 16.7%, P<0.001). Compared to non-recipient UTI patients, the symptoms of urinary irritation in recipient UTI patients were more common. There was higher percentage of neutrophil granulocyte (72.65% ± 1.90% vs 68.59% ± 0.73%, P=0.048), lower proportion of lymphocytes (17.73% ± 1.27% vs 21.28% ± 0.61%, P=0.037), and less platelets [(187.64 ± 10.84) × 10(9)/L vs (240.76 ± 5.26) × 10(9)/L, P<0.01] in recipients than in non-recipient UTI patients.

CONCLUSIONThese results indicate that the characteristics of UTI in kidney transplantation recipients and non-recipients patients are different.

Candida albicans ; isolation & purification ; Escherichia coli ; isolation & purification ; Female ; Humans ; Kidney Transplantation ; Klebsiella pneumoniae ; isolation & purification ; Male ; Transplant Recipients ; Urinary Tract Infections ; epidemiology ; pathology

OBJECTIVETo investigate antimicrobial resistance of Klebsiella pneumoniae and Pseudomonas aeruginosa isolates in fecal samples from rat-like animals.

METHODSRat-like animals were captured using cages around a hospital and the neighboring residential area between March and October, 2015. K. pneumoniae and P. aeruginosa were isolated from the fecal samples of the captured animals. Antimicrobial susceptibility test was performed according to the guidelines of Clinical and Laboratory Standards Institute (2014).

RESULTSA total of 329 rat-like animals were captured, including 205 Suncus murinus, 111 Rattus norvegicus, 5 Rattus flavipectus and 8 Mus musculus. The positivity rates of K. pneumoniae and P. aeruginosa were 78.4% and 34.7% in the fecal samples from the captured animals, respectively. K. pneumoniae isolates from Suncus murinus showed a high resistance to ampicillin, cephazolin, nitrofurantoin, piperacillin and cefotaxime (with resistance rates of 100%, 51.2%, 44.2%, 37.2%, and 23.3%, respectively), and K. pneumoniae isolates from Rattus spp. showed a similar drug-resistance profile. The prevalence rates of multidrug resistance and ESBLs were 40.9% and 10.7%, respectively. P. aeruginosa from both Suncus murinus and Rattus spp. exhibited the highest resistance rates to aztreonam (12.4% and 16.0%, respectively), followed by penicillins and fluoroquinolones. P. aeruginosa isolates were susceptible to cephems, aminoglycosides and carbapenems (with resistance rates below 5%).

CONCLUSIONK. pneumoniae and P. aeruginosa isolated from rat-like animals showed drug-resistance profiles similar to those of the strains isolated from clinical patients, suggesting that the possible transmission of K. pneumoniae and P. aeruginosa between rat-like animals and human beings.

Animals ; Anti-Bacterial Agents ; pharmacology ; Carbapenems ; pharmacology ; Drug Resistance, Bacterial ; Fluoroquinolones ; pharmacology ; Humans ; Klebsiella pneumoniae ; drug effects ; isolation & purification ; Mice ; Murinae ; microbiology ; Pseudomonas aeruginosa ; drug effects ; isolation & purification ; Rats

Animals ; Chickens ; microbiology ; Cross-Sectional Studies ; Enterobacteriaceae ; genetics ; isolation & purification ; Escherichia coli ; genetics ; isolation & purification ; Escherichia coli Proteins ; genetics ; Food Microbiology ; Klebsiella pneumoniae ; genetics ; isolation & purification ; Meat ; microbiology ; Multiplex Polymerase Chain Reaction ; Proteus mirabilis ; genetics ; isolation & purification ; Raw Foods ; microbiology ; Singapore ; beta-Lactamases ; genetics

Klebsiella pneumoniae liver abscess has a tendency to spread to distant sites early in the course of disease and to involve multiple organs synchronously. A 59-year-old male was admitted because of liver abscess accompanied by fever and abdominal pain. The patient underwent percutaneous catheter drainage and received intravenous antibiotics. Symptom relief was achieved after the treatment as well as marked reduction in the size of the abscess. Despite proper treatment of the liver abscess, however, patient developed multiple metastatic infections in a non-concurrent manner: left and right endophthalmitis, psoas abscess, and infectious spondylitis at 5, 23, 30 and 65 days after initial manifestations of liver abscess, respectively. Each infectious episode followed one another after resolution of the former one. For each episode of metastatic infections, the patient promptly underwent treatment with systemic and local antibiotics, interventional abscess drainage, and surgical treatments as needed. The patient fully recovered without sequelae after the use of intravenous antibiotics for an extended period of time. Herein, we report a case of K. pneumoniae liver abscess complicated with delayed-onset multiple metastatic infections.
Anti-Bacterial Agents/therapeutic use ; Ceftriaxone/therapeutic use ; Drainage ; Endophthalmitis/diagnosis/drug therapy ; Humans ; Injections, Intravenous ; Klebsiella Infections/complications/*diagnosis/drug therapy ; Klebsiella pneumoniae/isolation & purification ; Liver Abscess/*diagnosis/etiology ; Male ; Middle Aged ; Psoas Abscess/diagnosis/etiology ; Spondylitis/diagnosis/drug therapy ; Tomography, X-Ray Computed