ABSTRACT
Since 2011, the Republic of Korea (ROK) and United States (U.S.) have been collaborating to conduct inter- and intra-governmental exercises to jointly respond to biological events in Korea. These exercises highlight U.S. interest in increasing its global biosurveillance capability and the ROK’s interest in improving cooperation among ministries to respond to crises. With Able Response (AR) exercises, the ROK and U.S. have improved coordination among US and ROK government and defense agencies responding to potential bio-threats and identified additional areas on which to apply refinements in policies and practices. In 2014, the AR exercise employed a Biosurveillance Portal (BSP) to facilitate more effective communication among participating agencies and countries including Australia. In the present paper, we seek to provide a comprehensive assessment of the AR 2014 (AR14) exercise and make recommendations for future improvements. Incorporating a more realistic response in future scenarios by integrating a tactical response episode in the exercise is recommended.
Subject(s)
Australia , Biosurveillance , Disease Outbreaks , Exercise , Korea , Public Health , Republic of Korea , United StatesABSTRACT
BACKGROUND: Vancomycin-resistant enterococci (VRE) with vanA gene have been reported as a significant nosocomial pathogen. The vanA gene cluster (Tn1546) located on mobile DNA elements is known to be transferable from VRE to other enterococci. The purpose of this study was to investigate the genetic relationship between the vanA VRE strains isolated from hospitalizd patients and poultry. METHODS: Total 145 isolates, including 58 E. faecium, 12 E. faecalis, 3 E. casseliflavus, and 4 E. gallinarum from humans and 68 E. faecium from poultry, were studied. Antimicrobial susceptibility tests were done by disk diffusion or agar dilution methods and molecular epidemiological analysis was performed by pulsed-field gel electrophoresis (PFGE). The internal and structural regions of vanA gene cluster were analyzed by PCR fragment length polymorphism, restriction enzyme, and sequencing of Orf2D region and vanXY intergenic region. The point mutation at Tn1546 nucleotide position 8234 (G->T) within the vanX gene was screened with DdeI restriction enzyme. RESULTS: The antibiotic resistance patterns of human isolates were different from those of poultry. PFGE patterns revealed high heterogeneity. Three PCR fragment length patterns in the vanA gene cluster were found : (I) PCR amplicon of the same size as prototype (E. faecium BM4147) in 17% of human isolates and 100% of poultry ones; (II) PCR amplicon for vanXY intergenic region due to an insertion between vanX and vanY genes in 2.5% of human isolates; (III) the insertions in vanX-vanY intergenic and Orf2 regions in 81% of human isolates. The T type in vanX gene of human and poultry isolates was not found. CONCLUSION: Despite the diverse PFGE patterns, 81% of human and all of poultry isolates belonged to vanA gene cluster type III and I, respectively. These results indicate that the horizontal spread of vanA gene is occurring among genetically diverse strains of VRE in Korea.
Subject(s)
Humans , Agar , Diffusion , DNA , DNA, Intergenic , Drug Resistance, Microbial , Electrophoresis, Gel, Pulsed-Field , Korea , Multigene Family , Point Mutation , Polymerase Chain Reaction , Population Characteristics , PoultryABSTRACT
BACKGROUND: Enterococci are important cause of nosocomial infections. Recently, vancomycin-resistant enterococci (VRE) has been increasingly reported as significant nosocomial pathogens. Therefore, accurate identification of enterococcal species is a prerequisite step for the appropriate antibiotic treatment and epidemiologic surveillance. We wanted to know the usefulness of PCR method compared with Vitek automatic identification system. METHODS: Totally 105 isolates were identified on the species level by Vitek (GPI card and software version R06.1), methyl-alpha-D-glucopyranoside test, and PCR methods. RESULTS: Among 105 enterococcal isolates, 59 were identified as E. faecium, 11 E. faecalis, 6 E. gallinarum by Vitek. But 29 isolates (28%) were unidentified. Subsequently all of these isolates were analyzed by PCR, the results of which were as follows: 17 E. faecium, 5 E. casseliflavus, 7 E. gallinarum. Two isolates identified as E. gallinarum by Vitek were reidentified as E. casseliflavus by PCR and other methods for phenotypic characterization. CONCLUSOIN: PCR method was more accurate and sensitive than Vitek for the identification of enterococci species.
Subject(s)
Cross Infection , Epidemiological Monitoring , Polymerase Chain ReactionABSTRACT
ITSI-5.8S-ITSII rDNA region was amplified from the reference strains and clinical isolates with ITS1 and ITS4 primers. These primers amplified DNA fragments of 550 bp in Microsporum audouinii and Trichophyton violaceum, 700 bp in Microsporum gypseum, Trichophyton mentagrophytes, Trichophyton rubrum, and Trichophyton tonsurans, and 750 bp in Microsporum ferreugineum and Microsporum canis. The restriction enzyme patterns of PCR products digested with 13 restriction enzyme including PstI were distint among the genera, whereas identical in the same species. Examination of the ITS (Internal Transcribed Spacers)1 nucleotide sequence revealed that there was the genetic difference in each genera and species. Phylogenetic relationship among each species showed that the Trichophyton mentagrophytes was more closely related Trichophyton tonsurans than Trichophyton rubrum, and Microsporum gypseum was less related than Microsporum spp.
Subject(s)
Arthrodermataceae , Base Sequence , DNA , DNA, Ribosomal , Microsporum , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Ribosomes , TrichophytonABSTRACT
Dermatophytes infect the human hair, skin, nail and cause the dermatophytosis. The extracellular and intracellular proteinases of the dermatophytes commonly occur in the genus Trichophyton like T. rubrum, T. mentagrophytes, and T. granulosum. These enzymes play a prominent role in growth, multiplication and infection of the host tissue. Extracellular proteinases have been purified from the species of Trichophyton and Microsporum. We purified the proteinase partially from the culture filtrate of the Trichophyton tonsurans through Mono-Q and Superose 12 column and investigated its biochemical and enzymatic characters. The molecular size of the proteinase was estimated to be 41 kDa by SDS-PAGE. And pI was 3.2. The optimal temperature and pH for an enzymatic activity were 27C and 7.5, respectively. The purified porteinase degraded the keratin, bovine serum albumin, hemoglobin. The serine proteinase inhibitor like PMSF and DFP inhibited the proteolytic activity of the purified enzyme whereas the cysteinase inhibitor did not. These results demonstrated that the purified proteinase is a serine proteinase and can contribute the tissue invasion.
Subject(s)
Humans , Arthrodermataceae , Electrophoresis, Polyacrylamide Gel , Hair , Hydrogen-Ion Concentration , Isoflurophate , Microsporum , Peptide Hydrolases , Serine Proteases , Serine , Serum Albumin, Bovine , Skin , Tinea , TrichophytonABSTRACT
BACKGROUND: Dermatophytoses are infections of keratinized tissues, that is, the epidermis, hair and nails, caused by a group of specialized fungi, the dermatophytes. Laboratory diagnoses of dermatophytes such as Tricophyton, Microsporum and Epidermophyton are made by microscopic examination and in vitro culture but they are either time consuming of lacking specificity. OBJECTIVE: In order to develop and apply more rapid and precise diagnostic tests for fungal pathogens to facilitate the improved identification of dermatophytes, we investigated random amplified polymorphism DNA for classification and identification of dermatophytes. METHODS: Amplification reactions were performed in volumes of 5011 containing 10mM Tris-HCl(pH 8.3), 50mM KCl, 1.5mM MgCl2, 0.01% (w/v), gelatin, 200mM dNTP mixture, 50pM primer, Taq polymerase (0.025 units/ microliter), DNA 0.001 microgram/microliter. The optimal condition to. PCR was 2 cycles (denaturing 94 degrees C 2min, annealing 33 degrees C 2min, extension 72 degrees C 4min), 40 cycles, and extension (72 degrees C 10min). RESULTS: RAPD showed interspecies polymorphism in but it had identical patterns in intraspecies. CONCLUSION: It was confirmed that RAPD PCR analysis with optimal conditions is a fast, economical and reproducible method for identification and classification of dermatophytes isolates.
Subject(s)
Arthrodermataceae , Classification , Clinical Laboratory Techniques , Diagnostic Tests, Routine , DNA , Epidermis , Epidermophyton , Fungi , Gelatin , Hair , Magnesium Chloride , Microsporum , Polymerase Chain Reaction , Sensitivity and Specificity , Taq Polymerase , TineaABSTRACT
BACKGROUND: Trichophyton rubrum is the most common dermatophyte isolated from human and has ability to invade the tissues such as stratum comeum, nail and hair. The potential role of proteinases as virulence factors of F rMSrMm has been discussed at length. OBJECTIVE: As a first step towards assessing its virulence role, we report on the purification and characterization of proteinase from T. rubrum isolate culture filtrates. METHODS: An extracellular serine proteinase has been purified from culture filtrates of Trichophyton rubrum HP-9 by ultrafiltration, gel filtration chromatography, and affinity column chromatography. Azocoll and keratin azure were employed as the substrates of enzyme activities. Peak of proteolytic activity was analyzed by gelatin co-polymerized gel electrophoresis. RESULTS: The molecular weight of the purified enzyme was approximately exhibited to 14.0 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The optimum pH and molality of 14.0 kDa proteinase activity was 6.0 and 100mM, respectively. The activity was inhibited by serine proteinase inhibitor, phenylmethylsulfonyl fluoride (PMSF). The proteinase degraded gelatin, collagen type VI, and keratin from human epidermis but not hemoglobin. CONCLUSION: The 14,000 Mr extracellular serine proteinase purified from T. rubrum NP-9 culture filtrates has neutral pH optimum 6.0 and activities against gelatin, collagen type VI, and keratin.
Subject(s)
Humans , Arthrodermataceae , Chromatography , Chromatography, Gel , Collagen Type VI , Electrophoresis , Electrophoresis, Polyacrylamide Gel , Epidermis , Gelatin , Hair , Hydrogen-Ion Concentration , Molecular Weight , Peptide Hydrolases , Phenylmethylsulfonyl Fluoride , Serine Proteases , Sodium Dodecyl Sulfate , Trichophyton , Ultrafiltration , Virulence , Virulence FactorsABSTRACT
Enteroenteric anastomotic strictures of UGI tract are common and require treatment if significant obstruction occurs. We performed fluoroscopic guided balloon dilatation in 6 patients who had symptomatic stricture of gastrojejunostomy. The stricture was successfully resolved in 4 patients with benign stricture. But 2 patients with malignant stricture had recurrence of obstructive symptom 2 weeks later, and they required a stent. Asymptomatic balloon rupture was seen in one patient, but other procedural complications did not occur. We found that fluoroscopic guided balloon dilatation is an effective and safe method in the treatment of anastomotic stricture of gastrojejunostomy. We also found transient effect in malignant gastrojejunal anastomotic strictures, which required an interventional procedure, such as placement of a stent.
Subject(s)
Humans , Constriction, Pathologic , Dilatation , Gastric Bypass , Methods , Recurrence , Rupture , StentsABSTRACT
Enteroenteric anastomotic strictures of UGI tract are common and require treatment if significant obstruction occurs. We performed fluoroscopic guided balloon dilatation in 6 patients who had symptomatic stricture of gastrojejunostomy. The stricture was successfully resolved in 4 patients with benign stricture. But 2 patients with malignant stricture had recurrence of obstructive symptom 2 weeks later, and they required a stent. Asymptomatic balloon rupture was seen in one patient, but other procedural complications did not occur. We found that fluoroscopic guided balloon dilatation is an effective and safe method in the treatment of anastomotic stricture of gastrojejunostomy. We also found transient effect in malignant gastrojejunal anastomotic strictures, which required an interventional procedure, such as placement of a stent.