key: cord-0886355-bc3rob2c authors: nan title: Poster presentations date: 2017-09-30 journal: International Journal of Antimicrobial Agents DOI: 10.1016/s0924-8579(17)30342-4 sha: 9f621b3b10e1fcfa35c77ef2ce04427909e5aad4 doc_id: 886355 cord_uid: bc3rob2c nan susceptibility test of Salmonella species and Escherichia coli by VITEK (II). Results: We isolated 455 (20.5%, isolation rate) in 2012, 669 (31.4%) in 2013, 655 (30.8%) in 2014, 392 (25.7%) in 2015, 359 (22.0%) in 2016. Among the isolated bacteria, Staphylococcus aureus (740, 29.2%) was the most frequently identified pathogen followed by pathogenic Escherichia coli (646, 25.5%) Salmonella species (506, 20.0%), Bacillus cereus (223, 808%), Clostridium perfringens (219, 8.7%), Campylobacter species (188, 7.4%), Yersinia enterocolitica (8, 0.3%). Pathogenic E. coli, Salmonella species and Campylobacter species showed similar seasonality, showing a surge in the infection rate during the summer months. S. aureus, C. perfringens and B. cereus had no seasonal differences, while prevalence of C. perfringens was high particularly in 2013 and 2014. The antibiotic susceptibility patterns of isolated pathogenic E. coli and Salmonella species showed relatively high resistance to ampicillin (55.1%, 39.8%, respectively) and tetracycline (41.3%, 30.3%, respectively), while only 1 of Salmonella species showed resistance to imipenem. Conclusion: This study was based on the use of systematic surveillance of diarrheal patients, providing the geographic trend of bacterial pathogen and its antimicrobial susceptibilities. Keywords: bacterial diarrhea, antimicrobial resistance, Salmonella, E. coli, Gwang-ju city P1-BT10 Comparative genomic analysis of global clone 2 Acinetobacter baumannii clinical isolates during bloodstream infection L. Guo 1 , W. Yu 1 , B. Zheng 1 , Y. Xiao 1 *. 1 Background: Acinetobacter baumannii has gained significant attention because of the emergence of multidrug-resistant strains associated with outbreaks of hospital infections. It has been reported that the majority of such isolates belongs to two major global clones, GC1 and GC2, especially GC2 in Asia. But almost all the research focused on antibiotic resistance and epidemiology of these domain clones, little is known about the strategies this bacterium uses for pathogenesis. We used comparative genomics method to compare the genomic difference between blood and sputum A. baumannii isolates belonging to GC2. Methods: WGS was performed on 90 A. baumannii isolates from 45 individuals in a Chinese hospital, with one spit sample and one blood sample from the same patient. Core genome SNP were used to reconstruct the phylogeny of sequenced isolates, and the withinpatient SNP differences for every individual between bloodstream and sputum isolate was identified, at the same time, we recognized the genetic location of these SNPs and the common features of bloodstream infection isolates. Results: We observed 29% individuals with more than 300 pairwise SNPs, also had different ST types in blood-spit isolates, ST195 was the major ST type in blood isolates, but the ratio of ST195 in spit isolates was very low. We detected some candidant genes, such as capsular polysaccharide and lipopolysaccharide function related genes, had point mutations in blood isolates, which may be associated with bloodstream infections. Conclusions: Using WGS, we observed potential genes that are critical for the ability of this pathogen to cause bloodstream infections, this study lays the foundation for future research on A. baumannii genes that can be targeted to develop novel therapeutics against this human pathogen. Acinetobacter baumannii pRAY plasmids -An age-old, stealthy vehicle for the transmission of aminoglycoside resistance, overlooked S.S. Lean 1 *, C.C. Yeo 2 . 1 Saw Swee Hock School of Public Health, National University of Singapore, Singapore, 2 Biomedical Research Centre, Faculty of Medicine, Universiti Sultan Zainal Abidin, Malaysia Arising drug resistance in Gram-negative pathogens has led to an 'era of antibiotic resistance', forcing clinicians to face its greatest threatthe end of antibiotic use. Acinetobacter baumannii is a nosocomial pathogen that has rapidly acquired resistance in the past two decades. Advances in genome sequencing technology have provided a platform for the search of a panacea, resulting in a plethora of A. baumannii genome sequences in the databases. One of the interesting features of A. baumannii genomes is the presence of small plasmids (of around 10 kb or less), some of which encode antibiotic resistance genes. These plasmids tend to be overlooked in genomic analyses of A. baumannii that mainly focuses on larger conjugative plasmids and chromosomal features. Here, we aim to provide detailed analysis of the pRAY-type of small plasmids found in various Acinetobacter genomes and its possible role in the transmission of drug resistance. First isolated in South Africa from A. baumannii SUN, the pRAY plasmid confers resistance to aminoglycosides via the aadB gene. Subsequently, various pRAY derivatives ranging in size from ∼4-8 kb were found in other strains worldwide, most of which also harboured the aadB gene. Intriguingly, there were no potential replication initiation protein encoded within pRAY or its derivatives, suggesting that its replication may mirror that of the enterobacterial ColE1 plasmids that utilize the host RNA polymerase for its replication initiation. Another feature of this group of plasmids is the presence of mobA and mobC genes along with a putative origin of transfer (oriT), inferring the potential transmissibility of these plasmids. The discovery of a 4.1 kb plasmid, pALWED1.8 from a permafrost A. lwoffii isolate containing the oriT-mobC-mobA region plus an aadA7 aminoglycoside-resistance gene indicated that the oriT-mobC-mobA region is likely the backbone and common ancestor for this group of plasmids. This common ancestor would have acquired various other genes and spread to other Acinetobacter species throughout its evolutionary history. Procalcitonin rapid test: Adjunct biomarkers in early detection of bacterial sepsis with its prognostic value In the era of increased use of empirical antibiotic and culturenegative sepsis, positive bacteriological culture and non-specific infection markers remain inadequate for early detection of bacterial sepsis. This study aimed to determine the usefulness of rapid procalcitonin (PCT) in early detection of bacterial sepsis and its prognostic value. There were 132 patients recruited from intensive care unit and medical wards of clinical training centre from June to December 2016. This immunoassay uses a principle of immunochromatography and performed on day 1 and 5 of admission. Eighty microliter of whole blood was run in parallel to the negative and positive control. The PCT value will be obtained within 15 minutes and matched with the risk category of bacterial sepsis based on the manufacturer's recommendation including extremely high risk, high risk, moderate and low risk that has PCT value of >10ng/ml, 2 to 10ng/ml, 0.5 to < 2ng/ml, and < 0.5ng/ml respectively. Of 132 patients, 33 were excluded due to dengue, and remaining 97 were clinically diagnosed as having bacterial infection. Of 97 patients, 25.7% (n = 25) had extremely high risk of developing sepsis. While 12.4% (n = 12), 16.5% (n = 16) and 45.4% (n = 44) of patients had high, moderate and low risk towards sepsis. There were 13.4% (n = 13) death mainly due to septic shock and the highest number was found in the extremely high risks category, followed by moderate risk with increasing value of PCT of >5 ng/ml on day 5. Rapid PCT demonstrated a remarkable finding as it assists early detection of bacterial infection and guiding clinicians to commence empirical antibiotic therapy based on day 1 of PCT value. It also aids in predicting the prognostication in relation to severity of the infection by monitoring the PCT value. Audit of the Impact of Selective Reporting of Fluoroquinolone Susceptibility on Prescribing Habits in A Tertiary Hospital G. Foo*, S.Y. Tan, Z.C. Loh, W.T. Chung, K.L. Chew, N. Bagdasarian. National University Hospital, Singapore Background: Selective reporting of antimicrobial susceptibilities is one strategy commonly used among antimicrobial stewardship programs. Fluoroquinolone (FQ) susceptibility reporting was suppressed at our institution from 2011 until April 2016, during which time results were released only following special request from a clinician. After April 2016, FQ susceptibility was released as part of routine susceptibility reporting. The objective of this study was to evaluate whether the "unmasking" of FQ susceptibility results had any effect on the use of FQ for culture-directed treatment of an infection. Methods: We conducted a retrospective review of inpatient intravenous and oral FQ prescriptions for culture-directed therapy during a two month period prior to and after the "unmasking" of FQ in April 2016. FQ prescriptions were defined as appropriate if the only appropriate alternative was a carbapenem, or if it was the only appropriate oral agent prior to discharge. Overall FQ utilization was measured six months before and after this change in protocol. Results: There were 48 FQ prescriptions per month for culturedirected treatment before the change in protocol versus 80 FQ prescriptions per month afterwards. The percentage of inappropriate FQ use also increased from 49.5% to 60.6% before versus after the "unmasking". The most frequent indications for culturedirected FQ inappropriate use were for the treatment of urinary tract infections and lower respiratory tract infections in both time periods. The mean monthly fluoroquinolone utilization remained unchanged at 259 defined daily doses per 1,000 patient days before and after unmasking of FQ susceptibility. Conclusion: Selective reporting is a useful intervention in limiting the number of inappropriate fluoroquinolone prescriptions, which in turn can influence gram-negative resistance, MRSA and C.difficle rates in a hospital. strategy to decrease antibiotic resistance in hospital, particularly in intensive care setting. Methods: Antibiogram profile from Clinical Pathology Department of National Cardiovascular Center Harapan Kita Jakarta was retrospectively analyzed. Antibiotic restriction policy due to meropenem usage was started from November 2015 to November 2016. Meropenem resistance before and after its restriction was compared. Results: There were 335 and 177 isolates of bacteria had identified during 2015 and 2016, which is mainly from sputum specimen (79% in 2015 and 86% in 2016) . The most common bacteria in 2015 were Acinetobacter baumanii (28%), Pseudomonas aeruginosa (15%), Klebsiella pneumoniae (12%), and Serratia marcescens (5%). On the following year, the most common bacteria were Acinetobacter baumanii (18%), Klebsiella pneumoniae (14%), Serratia marcescens (12%) and Pseudomonas aeruginosa (10%). Antibiotic resistance rates of those bacteria to meropenem were 50% (blood culture) and 51% (sputum culture) before antibiotic restriction policy applied. These number were decreased to 33% (blood culture) and 28% (sputum culture) on the following year after antibiotic restriction policy. Conclusion: Meropenem resistance in pediatric cardiac intensive care unit was declining after antibiotic restriction policy. Keywords: antibiotic restriction, antibiotic resistance, meropenem, pediatric cardiac intensive care unit P1-CM08 Introduce the first triple Dot-ELISA diagnostic kit for Neospora caninum, IBR and BVDV M. Namavari 1 , S. Kamkar-salehi 2 Neospora caninum, Infectious Bovine Rhinotracheitis (IBR), and Bovine Viral Diarrhea (BVD) are the most important agent causes abortion in cattle worldwide. These infection agents cause reproductive disorders which can lead in economic loss in dairy and beef herds. Serological test including ELISA are calculated as the best diagnostic ways for Neospora caninum, IBR, and BVD. The aim of this study was to develop a multiplex Dot-ELISA kit for simultaneous detecting of three abortion agents: Neospora caninum, IBR, and BVD. The blood samples were collected from 184 cows with a history of abortion in the northern of Fars province. Then sera samples were evaluated by Multiplex Dot-ELISA and compare with commercial ELISA kits. Results showed that the rate of positive sera by use of commercial kits for Neospora caninum IBR and BVDV were 32. 98.9 and 64.1%, and by Dot-Elisa were 79.3 points 29.3 and 93.5 respectively. Kappa agreement assessment demonstrated that appropriate agreement between Multiplex Dot-ELISA results and ELISA kits, and McNemar test showed that both methods have the same efficacy for the identification of these abortion agents. The results of this study revealed that multiple Dot-ELISA kit, could be a simple and economical method for the simultaneous detection of three abortion agents in cattle. This experimentally kit is designed as the first step in designing a multiple kit for the diagnosis of infections in cows which could be introduced with complementary studies as a commercial kit in the future. Keywords: Multiplex Dot-ELISA, ELISA, Neospora caninum, IBR, BVD P1-CM09 Evaluate the role of the microbiological molecular Biology tests in the early detection of pathogens causing lower respiratory infections P.H. Van 1,2 *, V.D. Chien 2 , T.T. Kieu 2 , P.T. Huong 1 . 1 Nguyen Tri Phuong Hospital, 2 Namkhoa Co., 3 Background and aims: Based on the pathogenesis of the lower respiratory tract infections, the first specimen should be obtained from patients to detect the causative microbial agent is the sputum specimens. However, sputum is the contaminated specimen so that the big challenge must be overcome is to confirm the isolated bacteria is the pathogen, not the contaminated one. In addition, the most common pathogens of the lower respiratory tract infection are the fastidious bacteria requiring the immediate isolating on multiple media. Another specimen should be collected to detect bacterial pathogens causing pneumonia is the blood culture. The main challenge in the blood culture is the ratio of blood culture blood culture [+] in the diagnosis of pneumonia is often low and sometimes false positives because of contamination. So that blood cultures must be done at the right time on the appropriate blood cultures media. Serological and immunochemical test to detect the specific antibodies and antigens of the causative pathogens are the main solution for the detection of the pathogens that cannot be cultured routinely in most of the clinical laboratory like viruses and atypical bacteria, but these kinds of tests are often not sensitive and specificity enough (antigen and IgM detection) as well as not clinical relevant (IgG detection). Innovative and the most feasible solutions at present that can be able to detect microbial agents causing LRI are using the multiplex real-time PCR technique thank to its high sensitive and specificity, easily performed in the clinical laboratories due to the moderate investment costs and can implement automation. Methods and results: This solution has been evaluated by several studies were conducted in Nguyen Tri Phuong and Children No. 1 hospital, and now in REALS project. The received results have shown that multiplex real-time PCR capable of detecting pathogenic microbial agents with the sensitivity several times higher than the routine method. Model to implement this solution is called STREAMLINE REAL-TIME PCR with two basic devices: (1) The automatic DNA/RNA extraction machine using NK DNARNAPREP-MAGBEAD, and (2) the real-time PCR machine using kits including NK ARIbac real-time PCR for detection of community bacteria, NK ARIatypicalbac real-time PCR for detection of atypical bacteria, NK ARIvirus real-time PCR for detection of viral pathogen, NK HAPVAPbac real-time PCR for detection of nosocomial bacteria causing HAP and VAP bacterial pathogens. Conclusions: With this solution, the results of detection microbiological pathogens causing lower respiratory infection can arrive to the physicians timely, avoid the using of the empirical antibiotic treatment for longtime since the targeted antibiotic treatment can be done to the patients sort time after the clinical diagnosis. Keywords: Micro-organism pathogens causing pneumonia and lower respiratory tract infection P1-CM10 Production and evaluation of the kit using magnetic silica coated nano-iron beads to extract the nucleic acid from different samples P.H. Van 1,2 *, B.T. Thanh 3 , N.V. Quoc 2 , N.Q. Viet 2,4 , P.T. Huong. 1 Nguyen Tri Phuong Hospital, 2 Nam Khoa Co., 3 University of Medicine and Pharmacy, 4 Pham Ngoc Thach University of Medicine Introduction: PCR technique in Vietnam is being applied in many diagnostic laboratories. However, output quality varies between Background: In clinical practice, it is often difficult to distinguish true bacteremia from false bacteremia. The study aimed to evaluate the time to positivity (TTP), procalcitonin (PCT) level, and TTP/PCT ratio as reliable predictors for distinguishing true bacteremia from contaminated blood culture or intravascular catheter colonization. Methods: The study was retrospectively conducted in a university hospital in South Korea from January 2014 to December 2016. Subjects included febrile hospitalized adult patients (≥18 years) who had one or more positive blood cultures with available TTP and PCT data. The clinical findings of the patients were meticulously reviewed by an infectious disease physician. Positive blood cultures were categorized into true bacteremia and inconclusive bacteremia. True bacteremia was defined as having two clinically relevant positive blood cultures obtained on separate occasions. Inconclusive bacteremia was defined as a contaminated, single positive blood culture or catheter colonization. Results: During the study period, 172 patients were enrolled, including 129 (75%) for true bacteremia and 43 (25%) for inconclusive bacteremia. In a multivariate analysis, urinary tract infections [odds ratio (OR) 3.32, 95% confidence interval (CI) 1.03-10.72, p = 0.04] and shorter TTP [OR 0.97, 95% CI 0.94-0.99, p = 0.01] were independently associated with true bacteremia. In true bacteremia cases, TTP was significantly shorter for gramnegative bacteremia, compared to gram-positive bacteremia [16.0±13.9 vs 27.9±14.9, p < 0.001]. Based on primary site of infection, TTP was significantly shorter for urinary tract infections than for non-urinary tract infections [17.7±11.7 vs 23.7 ±17.1, p < 0.001]. In receiver operating characteristics curve analysis, PCT level (at the cutoff of [CO] 0.82 ng/mL), TTP (CO 20.5 h) , and TTP/PCT ratio (CO 36.5 h mL/ng) showed sensitivities of 73.6%, 72.9%, 82.2% and specificities of 62.8%, 65.1%, 74.4%, respectively. Conclusions: This study indicates that a shorter TTP or the presence of urinary tract infection may be reliable predictors of true bacteremia. In addition, TTP/PCT ratio could be useful in predicting true bacteremia. Keywords: procalcitonin, time to positivity, TTP/PCT ratio, bacteremia The qnrB gene is prevalent in Citrobacter freundii clinical isolates. However, there are no reports regarding the coexistence between the qnrB and bla VIM-2 genes. The aims of this study were to access the coexistence of these two genes and the potential of multidrug resistance transfer, and to characterize the mobile element involved. MICs of the antibiotics and an efflux pump inhibitor were determined in a clinical isolate, its transformant and site-directed mutant, according to guidelines of CLSI, by agar dilution method. Molecular characterizations of qnrB62 gene and its mobile element were performed by PCR amplification, DNA sequencing, PFGE and/ or Southern blot analysis. Coexistence of qnrB62 (encoding QnrB62 with Ser198Asn substitution, compared with QnrB5) with bla VIM-2 gene was detected in a Citrobacter clinical isolate resistant to fluoroquinolones and carbapenems. The reduced fluoroquinolone susceptibility is attributable to the qnrB62, a Thr83Ile substitution in GyrA and a Ser80Ile substitution in ParC, and efflux pump(s) other than QepA or OqxAB. The genetic context surrounding chromosomal qnrB62 was a novel complex class 1 integron (In1148::ISCR1::qnrB62) containing a unique gene array (bla VIM-2 -aacA4′-8-gucD) in a new class 1 integron (In1148). An 18 nucleotide deletion at the 3′ end of pspA (Δ18) upstream of qnrB62 and an inverted repeat region (IRR2) were detected in In1148::ISCR1::qnrB62, indicating past transposition events. The fluoroquinolone and carbapenem resistance associated with the In1148::ISCR1::qnrB62 integron could be related to a potential multidrug resistance horizontal spread that is worrisome worldwide. Stability of trimethoprim-sulfomethoxazole resistance genes in a strain of non-typeable Haemophilus influenzae Z. Mohd-Zain 1 *, S.Y. Mohamad 1 , N.K. Palanisamy 1 , N. Ahmad 2 . Background: Combination of trimethoprim-sulfamethoxazole (SXT) is commonly used in the treatment of Haemophilus influenzae infections. However, nowadays it is rarely used for treatment due to emergence of H. influenzae SXT-resistant strains. Resistance to trimethoprim is attributed to altered structure of dihydrofolate reductase gene while sulfonamide resistance is due to mutation in folP gene as well as, acquisition of sul2 gene. In this study, we investigated whether these resistance genes in a strain of nontypeable H. influenzae (NTHi) could be transferred to H. influenzae type B (Hib) by conjugation; and to determine if integrative and conjugative element (ICE) is involved in this process. Methods: Three types of mating procedures (filter-paper method, liquid-suspension method and agar-plate method) were carried out, in donor to recipient ratio volumes of 1:1 and 1:10. NTHi strain H152 resistant to SXT was the donor, while Hib strain H582, resistant to ampicillin, tetracycline and chloramphenicol was the recipient. Following overnight incubation, transconjugants bearing resistant genes were enumerated. PCR was used to detect the presence of attB in both strains, for common sequence of attachment site of ICE. Results: Repeated trials performed failed to produce transconjugants in all the mating methods. attB gene was detected in the Hib strain but absent in the NTHi strain was a possible reason that conjugative transfer of SXT-resistance genes did not occur. Conclusion: The genes responsible for SXT resistance in this NTHi strain are stable in its chromosome because this strain lacks the attB of an ICE for site-specific recombination to take place. Background: Moraxella catarrhalis is one of the important pathogen causing infectious diseases. The significance of Moraxella catarrhalis in community-acquired pneumonia is increasing with the introduction of pneumococcal conjugate vaccine and haemophilus influenzae type b vaccine. The aim of the study is to detect the antimicrobial susceptibility pattern and βlactamase activity characteristics of 178 Moraxella catarrhalis isolates collected from two county hospitals in China. Methods: Antibiotic susceptibility against 11 antimicrobials were tested using the E-test method or disc diffusion. The test results were interpreted with reference to the standards of European Committee on Antimicrobial Susceptibility Testing (EUCAST), Clinical and Laboratory Standards Institute (CLSI) and British Society for Antimicrobial Chemotherapy (BSAC). Detection of βlactamase activity was determined by the chromogenic cephalosporin nitrocefin (BR66A; Oxoid). Results: All of the isolates were susceptible to amoxicillinclavulanic acid. The susceptibility rate to meropenem was 100% according to EUCAST, as there was no breakpoints listed in CLSI and BSAC. The non-susceptibility rate to sulfamethoxazole-trimethoprim in the two counties showed significant difference no matter which judgment criteria was used, and isolates in Zhongjiang were more sensitive than strains in Youyang. The total positive rate of βlactamase was 99.4% (177/178 cases), as it was 99.0% (100/101 cases) in Youyang and 100% (77/77 cases) in Zhongjiang. Conclusion: Almost all of the Moraxella catarrhalis isolates produce β-lactamase. The isolates showed poor susceptibility to ampicillin and erythromycin, and high susceptibility rate to the third-and fourth-generation cephalosporins and amoxicillinclavulanic. There were some significant discrepancy between different antimicrobial susceptibility breakpoint criteria of Moraxella catarrhalis. Moraxella catarrhalis, Antibiotic resistance, β-lactamase, Children P1-GN04 Prevalence and antimicrobial drug resistance pattern of early urinary tract infection among renal transplant recipients in Nepal R. Shrestha. Tribhuvan University, Nepal Background: Urinary tract infection (UTI) is a major posttransplant bacterial infection in renal transplant recipients, with majority of infection encountered in first three month after transplantation. This is the first study conducted with the aim to determine the prevalence and drug resistance pattern of early UTI among renal transplant recipients at Human Organ and Transplant Center (HOTC), Nepal. Methods: A prospective descriptive study was conducted among renal transplant recipients transplanted between April to December 2016 at HOTC, and followed up until 3 month after transplant. Patients were screened for UTI, fortnightly for the first month and monthly for next two month and whenever sign and symptoms suggestive of UTI were present and questionnaires were filled up. Midstream urine and catheterized urine from newly inserted catheter were collected and processed according to the standard methodology. Both symptomatic and asymptomatic cases with bacterial colony count ≥10 5 cfu/ml were included. Combination disk method was applied for the detection of extended spectrum βlactamase (ESBL) and metallo βlactamase (MBL) producing isolates. Results: Total of 80 renal transplant recipients were included in the study, of which, 78% (n = 62) were male and 22% (n = 18) were female. Mean age of both genders was 35 years with standard deviation of 11 years. Total of 54 episodes of UTI were observed in 38.7% (n = 31/80) of transplant recipients, including 37% of male and 44% of female patients; (23/62) vs (8/18), (P value >0.05). Gram negative bacteria accounted for more than 90% of the UTI episodes. Klebsiella pneumoniae was the major bacteria, isolated in 42.6% (n = 23) of cases, followed by E. coli observed in 35.1% (n = 19) of cases. Other isolates were Pseudomonas aeruginosa, Enterococcus spp., Staphylococcus aureus and Citrobacter spp. All gram negative isolates were multidrug resistant. Increased level of resistance was observed against the most of drugs including; imipenem-44%, amikacin-57%, Gentamycin-61%, pipracillin tazobactam-83%, ofloxacin and ceftazidime 89% each, ceftriaxone-92%, cotrimoxazole -96%, amoxycillin and cefixime-98% each. Alarmingly high resistance to the carbapenem (78%), was observed among the isolates of Klebsiella pneumoniae. Polymixin B, colistin sulphate and tigecycline were only drugs with 100% susceptibility. The study showed high prevalence of gram negative isolates producing MBL (39.2%; n = 20/51) and ESBL (17.6%; n = 9/51). Carbapenem resistance due to carriage of MBL was alarmingly high among the isolates of Klebsiella pneumoniae, 73.9% (17/23). ESBL production was most common among the strains of E. coli, 42.1% (8/19). Conclusion: Early UTI is common among renal transplant recipient. It is further complicated by increased prevalence of multidrug resistance gram negative bacteria harboring ESBLs and MBLs. Proper infection control guidelines to fit our context, better adherence to policy and further study is urgently required for the better management and prevention of UTI with drug resistant bacteria. Characterization of integrons and antibiotic resistance to E. coli from cattle, sheep and goats in Iran Y. Tahamtan 1 *, T. Mostafaee 2 . 1 Microbiology Department, Razi Vaccine and Serum Research Institute, Shiraz Branch, Agriculture Research, Education and Extension Organization (AREEO), Shiraz, Iran, 2 Animal Science, Faculty of Agriculture Shahid Bahonar University of Kerman, Iran Background: E. coli infection is a major health problem especially among animal and children in developing countries. Moreover, E. coli is becoming resistant to commonly used antimicrobials in most parts of the world. Molecular characterization of class 1 integrons and prevalence of antibiotic resistance to E. coli from cattle, sheep and goats is reported. Methods: Swab stool specimens were collected from 250 cattle, sheep and goats from different parts of Fars province in Iran. Isolation and characterization were performed according to standard methodology including biochemical test for identification of E. coli and PCR for detection of the integron gene in isolates. Results: 173 E. coli strains were isolated, all of which were resistant to at least one antibiotic. 100% were resistant to amoxicillin, penicillin, tetracycline, ampicillin and co-trimoxazole, 92% to erythromycin and neomycin, 88% to gentamicin and 84% to amikacin. More than 50% were susceptible to sulfamethoxazole, enorfloxacin and cefalexin. Prevalence of integrons in the E. coli isolates was 34% and lack of antibiotic susceptibility to antibiotics was associated with its presence in bacteria. Discussion: E. coli may select a specific gene cassette through antibiotic selective pressure resulting in class 1 integron differences. Horizontal transfer through conjugative plasmids could be responsible for dissemination for class 1 integrons. Keywords: class 1 integrons, antibiotic resistance, cattle, sheep, goats, E. coli P1-GN06 Horizontal gene transfer of antibiotic resistance genes among Acinetobacter baumannii U. Leungtongkam 1 , R. Thummeepak 1 , K. Tasanapak 1 , S. Sitthisak 1 *. 1 Department of Microbiology and Parasitology, Faculty of Medical Science, Naresuan University, Phitsanulok, Thailand Conjugation is the horizontal gene transfer (HGT) that provides the most important mechanism to accelerate the spread of antibiotic resistance genes in Gram negative bacteria. Our study aims to determine gene transfer of extensively drug-resistant A. baumannii (XDR-AB) to A. baumannii isolated from environment using conjugation. Antibiotic susceptibilities were determined. Carbapenemase resistance genes (bla OXA-23 and bla NDM-1 ), aminoglycoside resistance gene (aphA6) and tetracycline resistance gene (tetB) were detected in all isolates. All drug susceptible Acinetobacter spp. recipients were selected for sodium azide resistance induction. Two isolates (NU013R, NU015R) which were resistant to sodium azide were used as recipient. Conjugation assays were performed to investigate the transfer of antibiotic resistance genes of 15 strains of XDR-AB to two sodium azide resistance isolates. Transconjugants were recovered on Mueller-Hinton agar plates containing sodium azide plus ticarcillin; plus tetracycline and plus kanamycin. Conjugation experiments demonstrated that resistance to ticarcillin and kanamycin could be transferred from only four XDR-AB donors to two sodium azide resistant A. baumannii. Conjugation frequencies (CF) for ticarcillin resistance varied from 2.0-3.7 × 10 −5 and CF of kanamycin resistance range from 2.3 × 10 −5 to 5.7 × 10 −7 . No transconjugant was detected on LBA with tetracycline. We found donor AB364 can transfer the bla OXA-23 gene to recipients, NU013R and NU015R, however the bla NDM-1 gene was negative in these transconjugants. The donors AB140, AB352 and AB405 can transfer kanamycin resistance gene (aphA6) to recipients, NU013R and NU015R. The REP-PCR profiles of donors, recipients and transconjugants were performed. The banding patterns of transconjugants were similar to the recipients. This work underlines how efficient the spread of the antibiotic resistance gene using conjugation could be among A. baumannii. Fecal carriage of carbapenem resistance Enterobacteriaceae among inpatients in a university hospital in Iran Objectives: Fecal colonization by carbapenem-resistant Enterobacteriaceae (CRE) could serve as a reservoir for transmission of these pathogens to clinical settings, which subsequently increases clinical infections. The aim of this study was to evaluate the prevalence and risk factors associated with CRE fecal colonization among inpatients. Material and Methods: Rectal swabs from 50 patients in a university hospital were collected. CRE screening was performed by using selective media. Carbapenemase production was detected by phenotypic tests. PCR assays were used to detect carbapenemases genes. Clonal relatedness was investigated by pulsed-field gel electrophoresis (PFGE). Results: The prevalence of fecal colonization was 56% (28/50). Overall, 41 CRE isolates were identified, of which 38 were carbapenemase-producers. Eleven patients (39.3%) were co-colonized with CRE isolates. ICU hospitalization, prior antibiotic therapy, and mechanical ventilation were significant risk factors. The bla OXA-48 was the most frequent carbapenemases followed by bla NDM-1 andbla NDM-7 enzyme. Nine carpapenemase producing Enterobacteriaceae (CPE) isolates co-harbored bla NDM-1 and bla . Also, six CPE isolates co-harboredbla NDM-7 and bla OXA-48 . We did not detect bla KPC , bla GES , bla IMP and bla VIM . PFGE analysis showed that E. coli clones were diverse, while K. pneumoniae categorize in 3 clusters. Cluster I was the major clone carrying bla OXA-48 and bla CTXM-15 genes. Conclusions: Our study as the first investigation in Iran showed CRE not only had high prevalence in fecal carriages, but also harbored varied antimicrobial resistance elements. Background: In Republic of Korea, carbapenemase producing enterobacteriaciae (CPE) infected patients were increased every year. Critical problem could be emerged in Korean public health system. From June 2015 to June 2016, two hospitals in Republic of Korea suffered from the outbreak of CPE. Those two hospitals were included in national surveillance whereby Korea Centers for Disease Control & prevention (KCDC) were completely controlled occurring of patients of CPE by intervention and control of transmission. Herein, we described medical and epidemiological characteristics of CPE patients in two hospitals and explained how control nosocomial transmission of CPE. Method: We reviewed the epidemiologic investigation reports for all cases about CPE outbreak in each hospital (hospital A: 36 patients, hospital B: 22 patients). We also reviewed medical records to collect data about the clinical course. The isolates of CPE were analyzed by National research Institute of Health in Korea for evaluation of carbapenemase. Based on the microbiologic results, we confirmed the outbreak of CPE. Pulsed-field gel electrophoresis (PFGE) was performed to clarify the epidemiologic correlation. Result: In hospital A, total 36 patients were involved in CPE (Klebsiella pneumoniae, KPC type carbapenemase) outbreak which occurred from surgical intensive care unit (SICU). 29 patients were transmitted in SICU, 5 patients were in medical intensive care unit (MICU), and 2 patients were exposed in general ward. We applied intensive CPE control strategies include active surveillance, patients isolation, emphasis of contact precaution to medical staffs, and preemptive isolation. About 11 months later, CPE outbreak in hospital A was terminated. In hospital B, 23 patients were in CPE (Klebsiella pneumoniae, NDM-1 type carbapenemase) outbreak which was originated from neuro-surgical intensive care unit (NSICU). Index case was occurred in SICU but he was admitted in neurosurgery (NS) department, because of cross-spread by doctors in NS, transmission of CPE in NSICU were generated. We closed NSICU for prevention occurrence of new CPE exposure. After intervention for control CPE outbreak, the outbreak was terminated within 1 month. Conclusion: We experienced CPE outbreak which occurred based on hospital setting. We successfully controlled that situation by strictly applying strategies of infection control. Characterization of copper resistance determinants among Acinetobacter baumannii clinical isolates R. Thummeepak 1 , A. Thanwisai 1 , N. Chantratita 2 , S. Sitthisak 1 . 1 Department of Microbiology and Parasitology, Faculty of Medical Science, Naresuan University, Phitsanulok, Thailand, 2 Department of Microbiology and Immunology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand Acinetobacter baumannii is a Gram-negative pathogenic bacterium which increases in prevalence of multiple-drug resistance strains have been documented worldwide. The copper resistance is one of the important mechanisms required for bacterial survival in environments and pathogenesis. This work aimed to characterize copper susceptibility, prevalence of copper associated genes and their expression profiles in A. baumannii clinical isolates. The copper associated genes were detected by the PCR assay in 102 clinical isolates of A. baumannii obtained from 3 hospitals in Thailand. The copper susceptibility testing was performed by the agar dilution method. The association of gene patterns and copper resistance was analyzed by using Fisher's exact test. The representative isolate was selected to study the effect of copper on the expressions of copper-related genes by using RT-qPCR. The prevalence of cueR and copR genes among tested strains was 100% (102/102) and 18.63% (19/102) respectively. The copper susceptibility study revealed that strains with high MIC copper (10 mM) were found in 24.51% (25 isolates), while 75.49% (77 isolates) of tested strains have MIC copper lower than 10 mM. We found that all 19 cueR + /copR + strains (100%) were resistant to copper (MIC = 10 mM) while only 6 of 83 (7.23%) of cueR + /copR − strains were copper resistant strains (P < 0.01). We also designed additional primers to detect structural genes of both cue and cop operons in a representative strain, AB 1521. We found that AB 1521 contains two structural genes (genes encoding for copper efflux pump and multi-copper oxidase) of cue operon, while cop operon contains six structural genes (genes encoding for CopB, CopA, CopC, copD, copper efflux pump and hypothetical protein). The results from RT-qPCR showed that all genes of cue operon were up-regulated at the micro-and millimolar levels of copper, while all genes of cop operon were over-expressed only at the millimolar levels of copper. This suggested that cue operon was intrinsic resistance system found in all analyzed strains and the cop operon was associated with copper resistance at the higher concentration of copper. The future study need to conduct in order to confirm the gene functions involved in copper resistance and pathogenesis. Keywords: Acinetobacter baumannii, copper susceptibility, copperassociated genes, RT-qPCR P1-GN10 Concentration gradient carbapenem exposure alters the plasmid copy number within nosocomial isolates of Escherichia coli harboring bla OXA-48 D. Paul 1 *, D. Dhar 2 , A. Bhattacharjee 1 . 1 Department of Microbiology, Assam University, 2 Silchar Medical College and Hospital, Silchar, India Background: The spread of bla OXA-48 carbapenemase underscores a potential threat and a grave concern for infection management and therapeutic options. However, it is not known how bacteria harboring this resistant gene will respond when carbapenem therapy is initiated to a patient. This study investigates the change in plasmid copy number of bla OXA-48 under exposure of different concentration of carbapenems. Methods: Clinical isolates of E. coli harboring bla OXA-48 within IncFrepB type plasmid were grown under exposure of imipenem, meropenem and ertapenem with concentrations of 0.5, 1, 2 and 4 μg/ml each. Plasmids were extracted and quantitative real time PCR was carried out to estimate the relative copy number of bla OXA-48 and fold change was normalized against housekeeping gene rpsel. Stability of bla OXA-48 was determined by serial passage without antibiotic pressure and curing of plasmid was done using different concentrations of SDS, acridine orange and ethidium bromide. Results: The copy number of bla OXA-48 was found to consistently higher as the concentration of ertapenem and meropenem was increased whereas in case of imipenem it was vice versa. Plasmids encoding bla OXA-48 were found to be highly stable and could be successfully eliminated after single treatment with SDS & is considered as the most efficient curing agent. The cured mutants became susceptible to carbapenems, quinolones and aminoglycosides. Conclusion: This study could establish that carbapenems effectively interfere with plasmid copy number, conjugative transfer, propagation and maintenance within a host, which in turn, help in bacterial adaptability in clinical settings and also significant for future infectious disease risk assessment and minimizing the selective pressure within the hospital environment. morbidity and mortality. The mechanisms of antibiotic resistance evolved in bacteria mainly include β-lactamase and efflux pump activities. Therefore, this study aimed to assess the β-lactamaseand efflux pump-mediated antibiotic resistance in laboratory-induced and clinically isolated antibiotic resistant strains of Salmonella Typhimurium. The antibiotic susceptibilities of S. Typhimurium ATCC 19585 (WT-ST), ciprofloxacin-induced antibiotic-resistant S. Typhimurium ATCC 19585 (CI-ST), and clinically-acquired antibiotic-resistant S. Typhimurium ATCC 19585 (CA-ST) were determined in the absence and presence of β-lactamase (clavulanate, sulbactam, tazobactam, and BLI-489) and efflux pump (CCCP and PAβN) inhibitors. The β-lactamase and efflux pump activity of WT-ST, CI-ST, and CA-ST were measured by nitrocefin hydrolysis and ethidium bromide (EtBr) accumulation assays, respectively. The susceptibilities of WT-ST, CI-ST, and CA-ST to most classes of antibiotics such as β-lactam, fluoroquinolone, aminoglycoside, and macrolide were considerably increased in the presence of βlactamase and efflux pump inhibitors. The β-lactamase activities were 11, 13, and 26 μmol/min/ml, respectively, for WT-ST, CI-ST, and CA-ST, which were increased to 13, 24, and 63 μmol/min/ml when exposed to ceftriaxone. The EtBr residues remained high in CI-ST (86%) and CA-ST (90%) when treated with PAβN. This study would provide useful information for better understating of the development of antibiotic resistance in association with β-lactamase and efflux pump activities and designing new antibiotic chemotherapy in combination with inhibitors. analysis. All ESBL-positive isolates and represented conjugants were subject to detailed characterization by whole-genome sequencing. Results: E. hormaechei and E. kobei were predominant among our isolates. MLST distinguished 46 STs with a polyclonal structure. ST591 was the most prevalent clone and detected in north China. Inter-and intra-regional clonal dissemination was identified by analysis of core-genome SNPs. Twenty-two isolates (35.5%) were ESBL positive, of which half were E. kobei. ESBL producers (n = 15) were exclusively classified as E. hormaechei and E. kobei, and bla CTX-M-3 was the most prevalent genotype (n = 10) detected within four genetic contexts. IncHI-type plasmids were prevalent in ESBLpositive isolates (13/22), especially in ESBL producers (12/15). All IncHI2-type plasmids carried ESBL genes and shared a similar backbone with a plasmid recovered from Salmonella enterica in Canada. Conclusions: E. kobei and E. hormaechei are the predominant reservoir of ESBL genes, and IncHI2-type plasmids largely contributes to the dissemination of ESBL genes in the community in China. The species distribution and species-dependent ESBL mechanism identified in our study suggests the necessity of tailored surveillance on the bacterium in the future. In vitro activity of tigecycline and comparators against multi drug resistant (MDR) Enterobacteriaceae in Africa the Middle East (AFME), and Asia-Pacific (AP) Conclusions: K. pneumoniae is a common pathogen that can express resistance to a broad spectrum of antimicrobials. The most active drugs (based on %S) were tigecycline, amikacin, and meropenem. Higher phenotypic-positive ESBL rates were observed among K. pneumoniae in AFME compared to AP region. Because of the propensity of K. pneumoniae to develop resistance and as resistance continues to spread, continued monitoring of this important pathogen through global and regional surveillance is warranted. isolates were identified and susceptibility tested locally using broth microdilution methodologies according to CLSI guidelines. CLSI breakpoint criteria were applied to define susceptibility rates. FDA breakpoints were used for tigecycline. Results: Susceptibility to agents by organism are provided in the following table. Conclusions: Tigecycline, meropenem, and amikacin were the only agents that consistently provided >90% susceptibility against the enteric species. However, given the propensity of susceptibility trends among these organisms to change over time, continued surveillance to monitor changes among key gram-negative bacilli from these countries is warranted. In patients with KP BSI, neutropenia, multiple organ dysfunction, respiratory failure, CnSKP infection, high APACHE II score, and tigecycline therapy were independently associated with higher mortality risk. Among patients whose APACHE II score was <15, higher mortality rates were observed in patients treated with tigecycline than in those treated with other antibiotics (45.3% versus 7.7%; P < 0.001). Conclusions: A significant increase in the incidence of CnSKP BSIs was observed during the study period, with a higher mortality rate seen in these patients. Exposure to carbapenems and severe illness were independent risk factors for the development of CnSKP BSIs, and tigecycline therapy resulted in a significant increase in mortality. Complete genome sequence and genomic characterization of two clinical Escherichia coli strains co-producing MCR-1 and NDM-1 X. Methods: Both isolates were subjected to EDTA-and phenylboronic acid (PBA)-carbapenem combined disk tests and characterized by phenotypic and genotypic methods. Results: Both isolates were confirmed to be class A IMI-1 carbapenemase producers by the presence of the synergy between imipenem and PBA followed by PCR and nucleotide sequencing. The bla IMI-1 was chromosomally encoded. Both of them gave an indistinguishable ERIC-PCR pattern. Neither extended-spectrum β-lactamase (ESBL) nor plasmid-mediated quinolone resistance (PMQR) was found in any isolate. They lacked OmpF but contained an identical OmpC variant. Both isolates remained susceptible to the third generation cephalosporins but were resistant to carbapenems with higher level resistance to imipenem (MIC of 16 mg/L) than those of ertapenem and meropenem. In addition, they were resistant to colistin (4 and 32 mg/L). Both patients were improved after antimicrobial therapy with either coamoxiclav or meropenem. Conclusion: Few reports of the IMI-1 enzyme may be due to its poor hydrolysis activity to the third-generation cephalosporins, suggesting the awareness of the unusual β-lactam resistance profile of the IMI producers. No statistically significant trends over time were found in any country; however, both Thailand and Vietnam showed a greater rate of ESBLs than other countries. Analysis of susceptibility patterns across countries was performed using data from the last 3 years combined to increase sample sizes. Susceptibility in Southeast Asia overall was low for most cephalosporins (<62%) and fluoroquinolones (<42%). Susceptibility was especially reduced in Vietnam (<40% and 22%, respectively) and Thailand (<55% and 32%, respectively), corresponding with the high ESBL+ rates in these countries. Of the studied agents, only amikacin, ertapenem, and imipenem demonstrated susceptibility >90% in each country. Conclusion: ESBL+ E. coli rates were high in Southeast Asia but overall relatively stable. Rates varied by country, with especially high levels seen in Vietnam and Thailand. As expected considering the high ESBL+ rates and co-resistance often found in ESBL+ isolates, susceptibility was low for cephalosporins and fluoroquinolones, limiting therapeutic options for UTI in Southeast Asia. Backgrounds: Opportunity for safe and successful treatment of carbapenem-resistant acinetobacter bacteremia (CRAB) bacteremia with appropriate antibiotic(s) (AA) is very limited in the real world. However, survivors without an AA does exist. We aim to review on the clinical features of these patients in order to characterize potential favorable prognostic factors for survival without an AA. Methods: All patients aged ≥18 years with CRAB bacteremia were retrospectively identified between Apr 2012 and Mar 2015 at five hospitals located in South Korea. Use of the AA was defined as administration of antimicrobial agent, to which the causative pathogen was susceptible, for more than 48 hours, within 5 days of the onset of bacteremia. Case (survivor) was defined as a patient who survived more than 28 days without use of any AA, while control (death) was defined as a patient who died within 5 days of blood culture without use of any AA. . Antimicrobial susceptibility was tested by the agar dilution method according to the CLSI guidelines. The following antimicrobials were tested: piperacillin, piperacillintazobactam, cefoxitin, cefotetan, imipenem, meropenem, clindamycin, moxifloxacin, chloramphenicol, and metronidazole. An inoculum of 10 5 CFU was applied with a Steers replicator and the plates were incubated in an anaerobic chamber for 48 h at 37°C. B. fragilis ATCC 25285 and B. thetaiotaomicron ATCC 29741 were used as controls. Results: Most of all isolates tested were susceptible to piperacillintazobactam, imipenem, and meropenem, as their resistance rates to these three antimicrobials were lower than 5%. Resistance rates to imipenem and piperacillin-tazobactam were 1% and 4%, respectively, for non-fragilis Bacteroides spp. organisms. However, much higher resistance rates were observed for piperacillin (38-69%), cefotetan (12-97%), and clindamycin (38-79%) for B. fragilis group, non-fragilis Bacteroides spp. and Parabacteroides spp. Overall, Prevotella, and Fusobacterium isolates are more susceptible than B. fragilis group organisms. Other anaerobic GNB were susceptible to most antibiotics tested. Conclusions: Piperacillin-tazobactam, cefoxitin, and carbapenems are highly active β-lactam agents against most of GNB. Continuous monitoring is necessary to detect the antimicrobial resistance pattern changes. Background: Despite the discovery of radiation energy hundred years ago, no significant research has been performed demonstrating its effect on bacteria. Here we reversed the carbapenem resistance in MDR Klebsiella pneumoniae using radaition. Methods: Overnight cultured bacteria were treated with 35 MeV proton particle energy with 20 Gy dosage. Replica plating was performed to screen the susceptible mutants. OMP profiles were determined and genomic DNA was sequenced using Ion Torrent PGM and PacBio-SMRT sequencing. Genomic and SNP analysis were performed using Geneious pro 8.1. Results: Among 100,000 colonies screened through replica plating, seven carbapenem susceptible K. pneumoniae mutants were obtained. The MIC for meropenem and ertapenem were changed from 4 and 8 µg/ml to 0.25 and 0.5 µg/ml, respectively. WGS analysis revealed that both the wild-type and mutant strains were found to have bla SHV-11 , bla TEM-B , bla OXA-4 beta-lactamase genes. However, there was an excision of 9 Kb nucleotide fragment from the plasmid associated with class 1 integron encoding bla CMY-10 gene in one of the susceptible mutants. Interestingly, neither inverted (IR) or direct repeats (DR) were detected, presumably due to the two transposase genes located at the 5′ end which did not leave any IR/DR marks. In addition, there were SNP's in two novel efflux pumps in all the mutants. OmpK35 and OmpK36 were found to be absent in both wild type and the mutants. Conclusion: The loss of function of two novel efflux pumps along with the deletion of 9 Kb nucleotide fragment, induced by the SOS response due to radiation, might have reversed the carbapenem resistance. CRISPR-Cas9 mediated knockout studies are in progress to understand the conclusive role of these targets. aureus (VISA) have been associated with treatment failure and the hVISA could not be detected by a simple disc diffusion method. Currently, there is no specific genetic marker for hVISA detection. Several genetic alterations were reported to be associated with the low-level vancomycin resistance in S. aureus. The aim of this study was to compare the isaA and rplN genes expression of VISA and hVISA with those of vancomycin-susceptible S. aureus (VSSA) isolates. Methods: A total of 75 MRSA isolates were classified as 32 VSSA, 28 hVISA and 15 VISA by population analysis profile with area under the curve (PAP-AUC). The expression levels of isaA and rplN genes in all isolates were determined by relative quantification real time RT-PCR and using 16s rRNA as the reference gene. The minimum inhibitory concentrations to vancomycin were determined by an agar dilution method. Results: The ranges of PAP-AUC for VSSA, hVISA and VISA groups were 0.12-0.87, 0.9-1.27, 1.35-3.91 with vancomycin MICs of 1-2, 1-3, 4->16 µg/mL, respectively. The VISA isolates showed significant higher relative expression of isaA than those of hVISA and VSSA (5.3 and 7.4 folds, respectively, p < 0.001), whereas rplN mean relative expression of hVISA was 2.2 folds higher than that of VSSA ( p = 0.037). Conclusion: This preliminary study reflects the potential role of the isaA and rplN expression as tool for the differentiation of hVISA and VISA from VSSA. Nocardia species are ubiquitous bacteria which can cause severe disseminated infection in humans. As their antibiogram is largely species-specific, susceptibility testing is not routinely performed for Nocardia species and sulphonamides remain the cornerstone of treatment. However with increasing sulphonamide resistance reported and geographical variability both in species distribution and in the resistance rate, there is a need to understand the local epidemiology and to detect any emerging resistance in the institution. This study aims to survey the distribution and antimicrobial susceptibilities of clinical Nocardia species received in a large tertiary hospital in Australia. Antimicrobial susceptibility testing was performed on 270 clinical Nocardia isolates received from January 2011 to December 2016 using the Sensititre™ RAPMYCOI microdilution panel (TREK Diagnostic Systems Ltd, Thermo Scientific). These were identified by partial 16S rRNA and/or secA1 sequencing and the panel of antibiotics tested were amikacin, amoxicillin-clavunalate, cefepime, cefoxitin, ceftriaxone, ciprofloxacin, moxifloxacin, clarithromycin, trimethoprim/sulfamethoxazole, doxycycline, minocycline, imipenem, linezolid, tobramycin and tigecycline. The three most frequently isolated Nocardia species were Nocardia nova complex (n = 80; 29.6%), Nocardia cyriacigeorgica complex (n = 61; 22.6%) and Nocardia brasiliensis (n = 52; 19.3%). Other species isolated included: Nocardia farcinica (n = 38; 14.1%), Nocardia psuedobrasiliensis (n = 8; 3.0%), Nocardia beijingensis (n = 6; 2.2%), Nocardia abscessus complex (n = 4; 1.5%), Nocardia transvalensis complex (n = 4; 1.5%) and other Nocardia species (n = 17; 6.3%) Of the tested strains, 25 (9.3%) isolates displayed resistance to trimethoprim/sulfamethoxazole (MIC 90 , 2/38 µg/mL); 40% of resistant strains were N. farcinica followed by N. pseudobrasiliensis (24%) and N. nova complex (16%) respectively. More than half of the isolates were resistant to imipenem (59.3%; MIC 90 , >64 µg/mL). Most isolates were susceptible to linezolid and amikacin with MIC 90 values of 4 µg/mL and <1 µg/mL respectively. Majority of resistance to trimethoprim/sulfamethoxazole is detected in Nocardia farcinica. Linezolid and amikacin remain as active agents and are good options in the treatment of Nocardia infection in this institution. Inducible clindamycin and methicillin resistant Staphylococcus aureus in a tertiary care hospital, Kathmandu, Nepal R.P. Adhikari* ,1 , S. Shrestha 1 , A. Barakoti 1 , R. Amatya 1 . 1 Department of Microbiology, Nepal Medical College and Teaching Hospital, Jorpati, Kathmandu, Nepal Background: Staphylococcus aureus, a frequent cause of infections continues to be an important nosocomial pathogen. Infections by it especially methicillin resistant ones are often difficult to manage due to its resistance to multiple antibiotics. Macrolide-lincosamide streptogramin B family of antibiotics is commonly used to treat such infections as an alternative to vancomycin. Method: A descriptive cross sectional study was conducted over the period of one and half year (November 2013-April 2015) in the microbiology laboratory of Nepal Medical College and Teaching Hospital, Kathmandu, Nepal to find the incidence of different phenotypes of MLS B resistance among S. aureus from clinical samples and their association with methicillin resistance. The study was done in 270 non-repeated isolates of S. aureus from clinical specimens ( pus, blood, urine, sputum and body fluids) from both gender and all age groups of patients attending NMCTH. Identification of S. aureus was first done using colony morphology on 5% sheep blood agar. Cream to golden yellow colonies with or without haemolysis were further identified using Gram stain, catalase test and coagulase test by standard microbiological techniques. Antibiotic susceptibility were studied by modified Kirby Bauer's disc diffusion method on Mueller Hinton Agar plates (12 cm diameter) using ampicillin (10 μg), cotrimoxazole (1.25/ 23.75 μg), ciprofloxacin (5 μg), vancomycin (30 μg), cephalexin (30 ug) and gentamycin (10 ug) discs. Cefoxitin (30 ug) for the detection of methicillin resistance and erythromycin (15 μg), clindamycin (2 μg) discs (Hi-media-India) at 15 mm apart were also used on same plate for the detection of inducible clindamycin resistance as per CLSI guidelines. Isolates with cefoxitin zone size ≥22 mm were considered methicillin susceptible and those with ≤21 mm were considered methicillin resistant. Isolates resistant to erythromycin and having a clindamycin zone ≥21 mm with a D-shaped zone were regarded as positive for inducible resistance ( i MLS B phenotype). Isolates resistant to erythromycin and susceptible to clindamycin were considered negative for D-test (MS phenotype) and those resistant to both erythromycin and clindamycin were regarded as constitutive resistance phenotypes ( c MLS B ). Isolates susceptible to both erythromycin and clindamycin were regarded as susceptible strains. Result: A total of 16,789 specimens (urine 7,970, blood 4,905, sputum 1,591, pus 1,504 and body fluids 819) were processed from both in-patients and out-patients. Of 270 isolates of Staphylococcus aureus (150 from male patients and 120 from female patients), 147 (54.4%), 60 (22.2%), 38 (14.0%), 20 (7.4%) and 5 (1.85%) were obtained from pus, blood, sputum, urine and body fluids respectively. Among the processed samples the highest positivity rate was found in pus sample (9.7.0%) followed by sputum (2.4%), blood (1.2%), body fluids (0.6%) and urine (0.2%). Among the antibiotics tested all the isolates were susceptible only to vancomycin. Gentamycin was still found to have better action as compared with other antibiotics. However, most of the isolates were resistant to commonly used antibiotics. Higher percentage of resistance to erythromycin and clindamycin among MRSA indicates that there is wide use of erythromycin and clindamycin for the treatment of staphylococcal infections in our set up. As this resistant patterns can be decreased by reduction in the use of macrolides, this study emphasizes the need to do likewise in our set up to reserve it as an alternative drug of choice for treating infection caused by MRSA. Low level of susceptibility of erythromycin resistant MRSA towards clindamycin means there is a least chance of clinical efficacy of clindamycin while treating erythromycin resistant MRSA infections as an alternative to vancomycin. These findings further emphasize the need of Dtest to be performed routinely in our set up to avoid clinical failure while using clindamycin as an alternative to anti-MRSA antibiotics like vancomycin and linezolid. Keywords: Staphylococcus aureus, MRSA, Inducible clindamycin resistance, Nepal Introduction: The major community-associated MRSA (CA-MRSA) strain in Korea, ST72-SCCmecVI, has been spreading in both community and healthcare setting, raising serious public health concerns. In this investigation, potential genotype-specific susceptibility profiles to host defense cationic antimicrobial peptides (HD-CAPs) were assessed using ST72-SCCmecIV and ST5-SCCmecII MRSA strains. Methods: We used 11 ST72-SCCmec IV and 9 ST5-SCCmec II MRSA clinical isolates, and assessed: (i) antibiotic resistance profiles; (ii) susceptibility to HD-CAPs (LL-37 and polymyxin B); (iii) relative net cell surface charge using FITC-labeled poly-L-lysine binding assay; (iv) transcriptional profiles of the genes involved in surface charge regulation in S. aureus (mprF and dltABCD); (v) spa type and superantigenic toxin type. Results: ST5-SCCmecII strains exhibited higher level of antibiotic resistance, especially multidrug resistance than ST72-SCCmecIV CA-MRSA strains. The ST5-SCCmecII strains also displayed higher resistance to LL-37 and PMB killing than the CA-MRSA strains, paralleling with higher level of surface positive charge linked to enhanced transcription of mprF and dltABCD among the ST5-SCCmecII strains. Conclusions: CA-MRSA ST72-SCCmecIV has been associated with lower mortality compared with ST5-SCCmecII MRSA strains, suggesting the CA-MRSA strains to be less virulent. In the current investigation, our data suggested that the lower pathogenicity of ST72-SCCmecIV MRSA compared with ST5-SCCmecII MRSA is associated with higher susceptibility to HD-CAPs, one of the critical elements in host innate immune system. Keywords: Methicillin-resistant Staphylococcus aureus (MRSA), Host defense cationic antimicrobial peptides (HD-CAPs), Genotype P1-GP05 Retrospective analysis of Staphylococcus lugdunensis susceptibility P. Rao 1 *, S. Mendis 1 , J. Chia 1 . 1 Tan Tock Seng Hospital, Singapore Background: Staphylococcus lugdunensis is known to be more susceptible compared to other coagulase negative staphylococci. Penicillin resistance rates of 12-40% have been reported and are due to beta-lactamases. Oxacillin resistance rates are less than 10%. We attempted to observe the temporal susceptibility trends for S. lugdunensis. Methods: In this retrospective analysis, we looked at antibiotic susceptibility data for S. lugdunensis from 2014 to 2016 at a tertiary care centre in Singapore. Susceptibility was routinely performed using either Vitek II or Kirby Bauer and interpreted using CLSI criteria (CLSI, M100). Results: A total of 440 isolates of S. lugdunensis were identified. Majority were from sites other than blood and urine (83%). Penicillin resistance was seen in 51% of isolates, and oxacillin resistance in 7.83%. Most isolates were susceptible to fluroquinolones, doxycycline and trimethoprim-sulfamethoxazole. Resistance to erythromycin (10%) and clindamycin (7.2%) was noted. Conclusion: In comparison to a study conducted in Singapore in 2005, which showed penicillin-resistance rates of 27%, there seems to be increasing penicillin-resistance among S. lugdunensis isolates. However, oxacillin and other antibiotics have remained largely susceptible. Preliminary study of RNA polymerase β subunit (rpoB) gene mutationsin vancomycin-susceptible and non-susceptible Staphylococcus aureus isolates from a Thai university hospital S. Wongthong 1,3 *, P. Tippayawat 2,3 , A. Chanawong 2,3 , R. Tavichakorntrakool 2,3 , C. Wilailuckana 2,3 , A. Lulitanond 2,3 . . Some VISA and hVISA isolates were reported to have rpoB mutation, which may play a major role in the development of vancomycin resistance in S. aureus. This gene encodes for a β-subunit of RNA polymerase, which is active in catalysis reaction. Some mutations in rpoB conferred transcriptional activity of the whole RNA polymerase. The most common mutation of rpoB in VISA was a substitution of H481Y. Therefore, we aimed to examine the rpoB mutations in MRSA isolates from a Thai university hospital. Methods: Ten MRSA isolates were classified for their vancomycin resistant phenotypes by using population analysis profile with area under the curve (PAP-AUC) method. The nucleotide sequences of rpoB from all isolates were determined and compared with that of the wild type isolate. Results: The 10 MRSA isolates included 3 VSSA, 2 hVISA and 5 VISA with their PAP-AUC ranges of 0.85-0.86, 0.92-1.05 and 1.46-1.91 respectively. Several isolates contained rpoB mutations. Two isolates of each VSSA, hVISA and VISA had amino acid substitution at H481N and/or S529L, similar to those previously reported in the MRSA isolates from several countries including Thailand. In addition, silent mutation was found in a VSSA, 2 hVISA, and 3 VISA isolates. Mutation of rpoB was not found in one VSSA isolate. Conclusion: Mutations in rpoB gene may be related to vancomycin resistance in S. aureus and should be further studied. Background: Vancomycin is mainly used for the treatment of patients with Staphylococcus aureus infections, and the decreased susceptibility to vancomycin is becoming increasingly common. Therefore, the finding of novel markers for the development of diagnostic tools for vancomycin-intermediate S. aureus (VISA) will be important. We applied a genome-wide association approach for discrimination between VISA and vancomycin-susceptible S. aureus (VSSA) using 79 whole-genome sequences (24 clinical isolates and 55 publicly available genome sequences). Methods: Vancomycin susceptibility was determined using agar dilution, broth microdilution, and E-test. Strains with MIC of >2 μg/ml in any of the three methods were defined as VISA. Genomic DNA was isolated from an overnight culture using the Wizard Genomic DNA Purification Kit and genome sequencing was performed using either the Illumina Miseq or PacBio RS II suquencer. We applied a quality-control to clean up raw reads using PRINSEQ version 0.20.3 and single-nucleotide polymorphism (SNP) calling performed using the samtools and bcftool. Functional annotations of the SNPs were carried out using an in-house developed perl script. We performed an association study using the PLINK software package and PPFS (Predict Phenotypes from SNPs) package add-on to kSNP3.0 and confirmed the SNPs associated with VISA phenotype using ClustalW multiple amino acid sequence alignment. Results: We obtained 46 SNPs which were associated with VISA phenotype. Among them, 27 SNPs showed substitution of nonsynonymous amino acid and 10 SNPs showed no amino acid change. We selected 5 candidates associated with reduced vancomycin susceptibility among 27 SNPs, based on the location and their function. Analysis using ClustalW multiple sequence alignment indicated that two novel SNPs were possible to distinguish VSSA and VISA, including iron transport SA_RS08075 (I63T) and KtrB (L212M). Conclusion: We found two novel markers to discriminate the VISA and VSSA, but additional study will be required to identify functional differences related to antibiotic resistance that may be resulted from these SNPs. Background: Small colony variant (SCV) is a slow-growing bacterial subpopulation, generally showing lower antimicrobial susceptibility than their wild-type (WT) strains. Here, we report SCV strains of vancomycin-resistant Enterococcus faecium (VREF), which has been very rarely reported previously. Methods: Two VREF isolates (2014-VREF-113 and 2014-VREF-234) showing SCV phenotype were isolated from rectal swabs of two hospitalized patients. MLST and PFGE were performed for molecular characterization of SCV and WT strains. We compared growth curve, stability of phenotype, antimicrobial susceptibility, and biofilm forming ability between SCV and WT strains. Results: In both VREF strains, MLST and PFGE patterns confirmed that SCV and WT strains were identical. Phenotype switching of 2014-VREF-234 was very stable even after 40 passages, while SCV phenotype of 2014-VREF-113 switched to the WT after 11 passages. There was no significant difference in antimicrobial susceptibility between SCV and WT strains of 2014-VREF-113 and 2014-VREF-234. However, biofilm formation of SCV strain was lower than that of WT in 2014-VREF-234. Conclusion: Two SCV rectal isolates showed the same antimicrobial susceptibility as WT strains. One SCV strain was very stable in multiple passages. Biofilm forming ability was not high in SCV strains and even one SCV showed lower ability than WT strain. All MRSA isolates were characterized by multilocus sequence typing (MLST). SCCmec typing and spa typing were performed for all isolates belonging to ST72. Results: Seven different STs were identified: ST5 (66.3%), ST72 (19.6%), ST239 (8.3%), ST254 (3.3%), ST1 (1.3%), ST30 (0.6%) and ST89 (1.3%). Before 2005, the most frequent ST was ST5 (61.6%), followed by ST72 (17.5%) and ST239 (11.6%). Between 2006 and 2008, ST5, ST72 and ST239 accounted for 43.3%, 30% and 23.3%, respectively. After 2011, ST5 and ST72 accounted for 55% and 43.3%, but ST239 accounted for only 1.7%. Among the ST72 MRSA isolates, the most predominant spa type was t324 (57%), followed by t148 (24%) and t664 (19%). Conclusion: Our results show that ST 72 MRSA has become increasingly isolated from clinical specimens of hospitalized patients in Korea, whereas ST239 MRSA has become decreased in prevalence over the past 1.5 decades. Identification of Helicobacter pylori in water sources in Makassar D. Nurul*, T. Uleng, I. Chaidir, I. Utami. University of Hasanuddin, Makassar, Indonesia Introduction: Helicobacter pylori (H. pylori) is a gram-negative, microaerophilic and spiral shaped bacterium that colonizes the human gastric mucosa. It is known that approximately 50% of the human population have been infected by H. pylori. The colonization of this microorganism is a major risk factor for peptic ulcer disease, gastric adenocarcinoma and mucose-associated lymphoid tissue (MALT). The prevalence of H. pylori reached more than 80% in developing countries andup to 22.1% in Indonesia, of which most infected ethnicities in Indonesia are Papua, Batak and Bugis. Previous studies have identified the relationshipbetween the prevalence of H. pylori infections with water sources in the community. Methods: This study is an observational analytic study with a cross sectional study design and purposive sampling technique. In this study, samples were taken from water sources in 14 slum districts in Makassar to find the presence of H. pylori and examined using Polymerase Chain Reaction (PCR). The synthesized DNA fragments used as positive control were based on the sequence of glmM gene represented a 294 bp band on electrophoresis gel, whereas distilled water were used as negative control. Results: Among 50 water samples taken in 14 slum districts in Makassar, there were no H. pylori detected byPolymerase Chain Reaction (PCR). Conclusion: We found that there were no contamination of H. pylori in water sources in slum areas in Makassar. Keywords: Helicobacter pylori, Polymerase Chain Reaction, water source P1-HI02 Surveillance study of Japanese encephalitis: an emerging infectious disease in India R. Kant 1 *, A. Yadav 2 . 1 University of Delhi, New Delhi, India, 2 Indian Council of Medical Research, New Delhi, India Japanese encephalitis, formerly known as Japanese B encephalitis is a disease caused by the mosquito-borne Japanese encephalitis virus (JEV). The (JEV) itself is a virus from the family Flaviviridae, which is a part of the Japanese encephalitis serocomplex of 9 genetically related viruses, some which are particularly severe in horses, and four known to infect humans including West Nile virus. The mosquitoes Culex tritaeniorhynchus and Culex vishnui are amongst the most important vectors of this disease. This disease is most prevalent in Southeast Asia, South Asia and East Asia. Japanese encephalitis viral activity has been widespread in India. The first evidence of presence of the virus dates back to 1952. First case was reported in 1955. Outbreaks have been reported from different parts of the country. During recent past (1998) (1999) (2000) (2001) (2002) (2003) (2004) , 15 states and Union Territories have reported Japanese encephalitis incidence. Annual incidence ranged between 1,714 and 6,594 and deaths between 367 and 1665. In this study, our group has tried to study the epidemiology of this particular viral disease and the data was collected and retrieved from the public databases as well as the local public hospitals across the infected regions. The epidemiological link between Methicillin-resistant Staphylococcus aureus (MRSA) colonised patients and the hospital environment H.M. Wong*, T.Y. Ng, Y.Y. Chua. Sengkang Health, Alexandra Hospital, Singapore Background: MRSA colonisation with subsequent infection contributes significantly towards patients' morbidity. Hospital environment is a potential route of transmission. Our hospital monitors MRSA acquisition rate by screening patients upon admission and prior to discharge. We noticed a rise in the MRSA acquisition rate in the hospital in February 2016. Method: We screened all patients in the hospital to assess the extent of the situation. Majority of them were from a male ward with 9 out of 17 patients acquiring MRSA in February 2016 during their stay. To establish how the patients acquire MRSA, we performed environmental sampling. We analyzed the positive isolates from patients and environment with Pulsed-field Gel Electrophoresis (PFGE). Results: The same strain of MRSA was found on 3 patients, 2 curtains and 1 day room chair; another 3 patients and 2 curtains' MRSA were genetically similar from this strain ( Figure 1 ). We emptied the ward for terminal cleaning and stepped up our daily targeted cleaning of high-touch surfaces. Curtains were all changed. Although the number of MRSA colonised patients had decreased in March 2016, there were still 6 patients colonised with MRSA in the ward despite the measures taken. We then took more cultures from the environment. We found that air-cooler filters in the same ward had identical MRSA strains from the first cluster in February 2016 ( Figure 2 ). After thorough cleaning of these filters and the environment again, only 2 patients (at ward's baseline) were found to be colonised in subsequent months. Conclusion: We confirmed the epidemiology link between MRSA colonised patients and hospital environment during a pseudooutbreak using PFGE. In the presence of stringent infection control measures, meticulous cleaning of high-touch surfaces in common areas is crucial in preventing the nosocomial transmission of MRSA via the hospital environment. Knowledge relating to use of antibiotics at a University in Sri Lanka N.P. Senanayake*, V. Navaratne, A. Balasuriya. Faculty of Medicine, General Sir John Kotelawala Defence University, Sri Lanka Introduction: Inappropriate use of antibiotics is a worldwide phenomenon occurring especially in the developing worldand it will lead to development of resistance to antibiotics. Objective: The aim of this study was to evaluate the knowledge relating to use of antibiotics among the pre-clinical and clinical students at the Faculty of Medicine, General Sir John Kotelawala Defence University, Sri Lanka. Methodology: A self-administered, pre-tested validated questionnaire from previous publications was used for data collection. The questionnaire based on the knowledge relating to use of antibiotics wasdistributed among all the medical students and the results were analyzed by comparing the knowledge of pre-clinical and clinical studentsat theFaculty of Medicine, General Sir John KotelawalaDefence University. The percentages were calculated and compared using z test for proportions. A p value of <0.05 was considered as statistically significant. Results: The age range of 159 participants was 17 to 26 years and 61.64% were males. There were 101 (63.52%) pre-clinical and 58 (36.48%) clinical students. The percentages of pre-clinical students who knew that the antibiotics should be used only for suspected bacterial infections, antibiotics should be started on doctor's prescription, importance of correct duration of antibiotic use and the importance of correct dosage interval of antibiotic use were 93.06%, 95.04%, 93.06% and 95.04% respectively. The percentage of clinical students who knew that the antibiotics should be used only for suspected bacterial infections, antibiotics should be started on doctor's prescription, importance of correct duration of antibiotic use and the importance of correct dosage interval of antibiotic use were 100%, 100%, 98.27% and 100% respectively. Significance (p) values of difference between two groups were 0.04, 0.08, 0.148 and 0.08 respectively. Of the 101 pre-clinical students 86 (85.14%) knew that the frequent and inappropriate antibiotic use could lead to development of antibiotic resistance. All the clinical students (100%) knew that frequent and inappropriate use of antibiotics would lead to development of antibiotic resistance (χ2 = 9.51, df = 1, p = 0.002). Conclusion: The knowledge relating to use of antibiotics was better among the clinical students in comparison to pre-clinical students. Background: Control and prevention of Methicillin-resistant Staphylococcus aureus (MRSA) transmission in healthcare remains a significant challenge worldwide. One of the main emphasis is on promoting hand hygiene practices among healthcare workers. However, the role of patients' contacts in nosocomial transmission of MRSA is unclear. The aim of this study is to evaluate contact Methods: This was a prospective observational study of 35 MRSApositive patients and 15 MRSA-negative patients admitted to our institution from March to December 2016. All episodes of patientto-patient contact, patient-to-environment contact and hand hygiene practices were observed and recorded. Associations between key exposure variables with contact episodes and hand hygiene practices were analyzed. Results: In all, there were 7 patient-to-patient contact episodes and 3442 environmental contact episodes observed in 289 observation hours. Median contact episodes per subject in patient zone and common healthcare zone were 49 (20-86) and 1.5 (0-6) respectively. Common point of contacts were bedrails, bedside table and toilet door handles. There were no statistical significant differences in contact patterns between MRSA-positive and MRSA-negative patients. Among patients who were functionally independent or assisted, 57% of MRSA-positive patients had >5 contact episodes in healthcare zone compared to 36% of MRSA-negative patients. Of these, only 38% of MRSA-positive patients performed hand washing compared to 64% among MRSA-negative patients. Conclusion: Direct patient contact and shared environment between patients and infrequent hand hygiene practices among MRSA patients could play a role in nosocomial transmission of MRSA. Introduction: Antimicrobial resistance (AMR) is an emerging problem in developed and developing countries. In the Philippines, MDH has been actively joining the global effort to fight AMR since 2013. In this study, we aimed to assess the initial impact of ASP on the prevalence of MDROs and antibiotic consumption measured as defined daily dose. Methodology: Review of surveillance data and antibiotic usage was done from 2011 to 2016. All specimens positive for K. pneumoniae (KPC), E. coli & K. pneumoniae (ESBL), and Methicillin-resistant S. aureus (MRSA) were examined. Prevalence rate was computed for the MDROs per year. Age, sex and length of hospital stay were gathered. Utilization of Cefepime, Ertapenem, Meropenem, Imipenem, Vancomycin, Pip-Tazo, Levofloxacin, and Ciprofloxacin was computed as DDD/1000 patient-days. Difference between the antibiotic consumption of before and after program implementation was evaluated using T-test. Spearman's correlation was used to test the relationship between resistance and antibiotic use. Results: The mean age of patients was 62. Most of the patients were male (58%). Patients in ICU have a longer length of stay than the patients in the 5A Station. Among the isolated MDROs, 39.71% (135 of 340 isolates) was K. pneumoniae (ESBL+). Pip-Tazo (25.04 DDD/ 1000 patient-days) and Meropenem (13.86 DDD/1000 patientdays) have the highest consumption among the monitored and restricted antibiotics, respectively. There is a strong correlation between the % resistance of E. coli and use of Cefepime (r = 0.7902, P = 0.0022). Conclusion: Increased utilization of Cefepime is correlated with the resistance in E. coli against this agent. Since the ASP was recently implemented, the true impact of ASP may not be evident as of the time of evaluation. Moreover, although subjects with latent TB infection (LTBI) are asymptomatic and are not infectious, they may eventually develop active disease. Thus, the screening and treatment of LTBI is important for a fundamental tool of TB control programs for HCWs. The objective of this study was to assess the prevalence and epidemiologic characteristics with LTBI among HCWs at a hospital in Seoul, South Korea. Methods: This cross-sectional study of HCWs was conducted between 2016 and 2017. Participants were administered a questionnaire on demographics. We excluded HCWs with history of TB or LTBI treatment in this study. The QuantiFERON ® TB-Gold (QFT) assay was performed for the screening of 740 HCWs who worked at Konkuk University Medical Center in Korea. Epidemiologic risk factors with LTBI were identified by logistic regression analysis. Results: The prevalence of positive QFT was 10.9% (81/740). There was no significant association between risk factors for sex, working in the medical domain and employment type and positive QFT. However, working in medical domain tended to be more positive QFT than working in surgical domain (7.6% vs. 3.4%, P = 0.075, OR 1.6, 95% CI 0.952-2.570). The higher age (≥40 years old) was significantly associated with positive QFT (OR 3.7, 95% CI 2.682-8.401, P < 0.001). All QFT positive HCWs were referred for chest Xray and examination by pulmonology specialists. None were found to have active TB, and were thus diagnosed with LTBI. Conclusions: Our findings suggest that the prevalence of LTBI in HCWs may be relatively high. The risk of LTBI increases with longer working experience. As a result, active serial screening for LTBI is needed for HCWs. Introduction: Improper practices and lack of knowledge by food handlers are contributing factors for the spread of foodborne outbreaks. This study aimed to explore the knowledge, attitude/ practices of food/hand hygiene and observation of hand washing practices among food handlers at the General Sir John Kotelawala Defence University (KDU) and the KDU guest house. Methodology: A descriptive cross sectional study was done among 103 food handlers who are involved in food production, possessing, distribution at the General Sir John Kotelawala Defence University and the KDU guest house. Information gathering method using a pretested validated questionnaire and direct observations were used for the data collection. An association was tested between hand/food hygiene knowledge/attitude and category of service (chefs, cooks, kitchen assistants, waiters and others including food distributing staff ), level of education (completed their primary education up to grade 5, completed GCE O/L and completed their GCE A/L) and duration of service (worked more than 5 years, for 1 to 5 years and for less than 1 year). Results: Of the 103 food handlers, 86.9% had excellent knowledge on food hygiene. Category of service and level of education of food handlers were related with knowledge /attitude of food hygiene ( p < 0.05). However, there was no significant difference between the knowledge of food hygiene with the duration of service of the employees ( p > 0.05). Knowledge and attitude of hand hygiene was excellent in all the food handlers (>86%) except the knowledge of 8 steps of hand washing techniques ( p > 0.05). Through direct observations, it was observed that food handlers' hand washing practices significantly related with the place of work (KDU and KDU guest house) and category of service (p < 0.05). Conclusions: Most of the food handlers at the General Sir John Kotelawala Defence University have good knowledge, attitude and practices regarding food and hand hygiene. Further educational and training programs will be necessary to improve their knowledge attitude and practices of food and hand hygiene. Background: Active surveillance culture of carbapenem-resistant Enterobacteriaceae (ASC-CRE) is recommended in high-risk settings. We aimed to investigate the effect of AST in patients admitted to an intensive care units (ICU) and the risk factors for CRE acquisition. Methods: We prospectively conducted ASC-CRE of rectal swabs for all patients admitted to ICU during a 5.5-month at a tertiary hospital in South Korea. The ASC-CRE was done at admission and was re-tested weekly. ASC-CRE was performed by CDC methods. A PCR assay was performed to detect 5 carbapenemase genes (NDM, KPC, VIM, IMP, OXA). Results: A total of 294 patients were admitted during the study period. CRE acquisition rate was 4.4% (13/294); 46.2% (6/13) were carbapenemase-producing Enterobacteriaceae (CPE). No invasive infection was found. The most common species were Klebsiella pneumoniae (76.9%, 10/13) and 5 KPC and 1 NDM genes were detected. In CRE-positive patients, in-hospital-mortality were higher (P = 0.002). Multivariate analyses showed prior exposure to healthcare facilities (outpatient visits, admission or surgery in the preceding 1 year) (aOR 13.9; 95% CI 1.6-117.1), co-colonization of multidrug-resistant organisms (MDROs) (aOR 4.9; 95% CI, 1.3-17.7) and the length of admission to CRE detection ≥10 days (5.6; 95% CI 1.5-20.4) were independently associated with CRE acquisition. Conclusion: Using ASC-CRE, CRE acquisition rate was 4.4%. Prior exposure to healthcare facilities, co-colonization of MDROs and the length of admission to CRE detection ≥10 days were risk factors for CRE acquisition. Background: The national surveillance system has been proved to reduce device-associated infection (DAI) through several studies and many countries have already applied it. Therefore, we investigated to time trend of DAI rates and factors associated with changing trend for 10 years through the Korean National Healthcare-associated Infections Surveillance System (KONIS). Methods: We investigated DAI rates from 2006 through 2015 in 190 KONIS participating intensive care units (ICUs) in 107 hospitals. DAI rates were calculated as numbers of infections per 1,000 devicedays. Data associated with ICU characteristics were collected and mixed linear regression analysis was used for statistical analysis. Results: From 2006 to 2015, rate of ventilator associated pneumonia (VAP) significantly decreased from 3.48 in 2006 to 1.00 in 2015 ( per 1,000 ventilator-days, F = 27.62, p < 0.0001). Also, rates of catheter associated urinary tract infection (CAUTI) and C-line associated blood stream infection (CLBSI) significantly decreased from 1.85 to 0.88 ( per 1,000 catheter-days, F = 10.14, p < 0.0001) and from 3.40 to 2.20 (per 1,000 catheter-days, F = 14.17, p < 0.0001). In subgroup analysis, rates of VAP, CAUTI and CLBSI were significantly decreased regardless of organizational and institutional characteristics of ICUs. Conclusion: In Korea, all of the DAIs have shown a significant reduction in the last 10 years, that might be associated with participating surveillance in the KONIS. We will need to encourage participation of more hospitals. Keywords: Device-associated infections, year-wise trend, surveillance, intensive care units P1-HI12 Antibiograms in South Indian Hospitals: myth or reality? Dr. D. Sureshkumar 1 *, Dr. K. Divya 2 , Dr. S. Soujanya 3 . 1 Apollo Hospitals, Chennai, 2 SIMS Hospitals, Vadapalani, Chennai, 3 Objectives: Periodic analysis and reporting of antimicrobial susceptibilities (antibiogram) have effectively improved antibiotic prescribing practices in the hospital. International societies recommend institutional antibiograms are helping in better implementation of antibiotic stewardship activities in the hospital. Little is known about how Indian hospitals compelling the antibiograms. The aim of the study was to study describe the key features of hospital antibiogram in South Indian Hospitals. Methods: Using cross-sectional research design, purposive sampling of accredited hospitals across South India was conducted using a structured questionnaire prepared based on latest CLSI antibiogram guidelines. Content validity of survey questionnaire was carried out by infectious disease physician & postdoctoral fellows. Survey questionnaire completed by consenting respondents (infection control team member or microbiology laboratory personnel) through either face-to-face interaction or through telephonic interview were taken for analysis. Data were compiled using descriptive statistics. Results: 18 of the 25 contacted respondents completed the survey. The other parameters surveyed were shown below in Table 1 . contributory. Beginning in July through September 2016, an increase in SSI's in Obstetrics & Gynaecology (OG) was noted. Our aims were to evaluate SSI cases for potential causes and clustering, measure peri-operative antimicrobial appropriateness, develop interventions to reduce SSI, and improve appropriate perioperative antimicrobial use. Methods: Data from SSI cases were collected and described. Subsequently, employing Plan-Do-Study-Act (PDSA) methodology, results were discussed and used to develop interventions together with stakeholders, and structural changes were suggested or implemented. Repeated reviews and changes were performed, and measurable processes were tracked in addition to SSI trends. Results: We confirmed an increase in SSI from baseline of 0.7% to 1.8%. There was no temporal or spatial clustering, but 33% were obese, 97% received peri-operative antibiotics within 60 min of incision, 76% received appropriate type/ dose of antibiotics, 55% received chlorhexidine-alcohol as surgical skin antisepsis, while 75% of obstetric cases occurred >14 days post-operatively, suggesting multiple possible causes. Working with multiple stakeholders, a bundle of 7 interventions were developed as best practices for implementation. We also rectified a 'downtime' form used by junior doctors who were prescribing peri-operative antibiotics erroneously. By end March 2017, total SSI rates had decreased to 0.4% from 1.8% (see Figure 1 ). All SSI patients received timely peri-operative antibiotics, 86% received chlorhexidine-alcohol as surgical skin antisepsis, though only 57% of SSI cases received appropriate type/ dose of peri-operative antibiotics. Conclusion: Continuous engagement of all key stakeholders are critical in implementing change by ensuring buy-in at all times. Improvements in outcomes often require both process as well as structural improvements. For some outcomes, a bundle of measures may be more effective than any intervention on its own. Current epidemiology revealed that there was diversity in distribution of fungal isolates from one centre to another, as it may influence the clinical practice in the management of mycoses. This study was undertaken to describe the distribution of fungal isolates in the new clinical training centre, Malaysia. Ninety-three fungal isolates from 92 specimens were detected on both saboraud dextrose agar and brain heart infusion media from January 2014 to March 2017. Identification of all isolates was performed by assessment of its colonial morphology, microscopy examination and VITEK-2. Majority of the fungi were isolated from nail (n = 32; 34.7%) followed by skin scrapping, blood, bronchioalveolar lavage and tissue biopsy respectively [31 (33.7%), 13 (14.1%), 13 (14.1%) and 3(3.26%)]. Commonly isolated fungi were consists of non-albicans candida followed by Candida albicans, Fusarium spp., Trichophyton spp., Aspergillus spp., Trichosporon spp., and Cladosporium spp. [19; (20.4%), 16 (17.2%), 15 (16.1%), 10 (10.8%), 9 (9.67%), 6 (6.45%), and 5 (5.37%)]. Sporothrix schenkii, Rhizopus spp. and Penicillium spp. contributed 2.15% of each of the isolated fungi while, Verruconis gallopava, Cryptococcus laurentii, Bipolaris spp., Curvularia spp., Acremonium spp., Chaetomium spp., and Microsporum spp. added only 1.07% to the distribution. Candida albicans remains the most common isolated fungi followed by Fusarium spp. and Trichophyton spp. All three isolates were mainly isolated from blood, skin scrapping, and nail. Emphasis on the need to have continuous surveillance regarding distribution of fungal isolates need to be carried out, as these will provide a resourceful information on the commencement of empiric antifungal therapy. The aim of this study was to produce fluconazoleloaded liposomal nanoparticles, to analyze their physicochemical properties and to compare their antifungal effects with the free fluconazole drug in vitro against the fluconazole susceptible and resistant Candida species isolated from patients. Six common candida species including C. albicans, C. parapsilosis, C. tropicalis, C. glabrata, C. krusei and C. guilliermondii were tested. The Liposomal nanoparticles were prepared using thinlayer hydration method and soybean lecithin, cholesterol, and fluconazole at a ratio of 10:1:1. The nanoparticles were analyzedin terms of size, poly dispersity index, zeta potential, morphology, entrapment efficiency of drug and the amount of drug released. To investigate the antifungal effects of liposomal nanoparticles and compare them with the free form of fluconazole, we used Broth Microdilution as described in CLSI M27-A3. The results were analyzed using Student's T-test and indicated the greater antifungal effects of the liposomal nanoparticles containing fluconazole than the normal form of the drug. It was shown that MIC of Fluconazole was put in the range of sensitive species after exposure with the Fluconazole Nanoliposomal in most Fluconazoleresistant Candida species except for the krusei species. Objectives: Candida bloodstream infections (BSIs) are associated with significant morbidity and mortality in patients with hematological malignancies. However, the issue of whether central venous catheters (CVCs) should be removed remains controversial, especially in neutropenic patients. Methods: This was a prospective observational study of adult patients with hematological malignancies developing Candida BSIs who had a CVC in place from 2011 to 2014. CVC removal was preferred in all non-neutropenic patients and in neutropenic patients when the source was presumed to be the CVC or who failed to show Candida clearance from the bloodstream despite appropriate antifungal therapy. Results: Of 40 patients with Candida BSIs, 24 (60%) were neutropenic and 16 (40%) non-neutropenic. Overall, the CVC was removed in 27 (68%) patients at any time points. The CVC was less frequently removed in neutropenic patients than in non-neutropenic patients (54% vs. 88%; P = 0.040). There was no difference in the mortality rates at 6 weeks following CVC removal or CVC retention (33% vs. 54%; P = 0.305). Independent predictors for the 6-week mortality were refractory neutropenia and Pitt bacteremia score. There was no difference of demographic and clinical characteristics between the CVC removal and retention groups. The 6-week mortality in neutropenic patients with Candida BSIs did not differ between the CVC removal and retention groups (54% vs. 55%; P = 1.000). (74) oral Candida isolates collected from T2DM patients in Misrata, Libya were characterised using the VITEK 2 Compact system. Results: Prevalent species included C. albicans, C. glabrata, C. dubliniensis, C. krusei, C. tropicalis, C. sake, C. kefyr, C. guilliermondii, C. parapsilopsis, C. membranifaciens and C. magnoliae. Drug susceptibility showed an emerging resistance across representatives of all species for which breakpoints were available, with the exception of C. parapsilopsis. Although there are no established interpretative breakpoints for these species, three C. sake isolates and the C. membranifaciens isolate also had high MIC values for fluconazole. The tested isolates were found to be largely susceptible to caspofungin and micafungin. All C. albicans isolates were susceptible to the echinocandins, amphotericin B and 5-flucytosine. Resistance to more than one drug class was seen in C. dubliniensis, C. glabrata and C. krusei isolates. Conclusion: Although the susceptibility results for the echinocandins were encouraging, resistance against the azoles was apparent and should not be ignored. This was especially so in the case of fluconazole, which is often the only locally available antifungal drug for the treatment of disseminated candidiasis. Currently, the live vaccine Toxovax is produced from attenuated variety of Toxoplasma gondii. The aim of this study was to evaluate the immunogenicity of live variety of T. gondii which produced through serial passages in cell culture. Twenty-four Balb/C mice were randomly divided into three groups. Group I was the negative control and received cell culture media groups II as positive control and III were inoculated with, acute, and Razi strains of T. gondii, respectively. All mice were assessed daily and potential mortality was recorded. Twenty-eight days later blood sample were collected from all groups for ELISA and then all mice were challenged with injections of acute strain of T. gondii. The results showed that in the control group mortality was started from day 8 and up to14 days after challenge all mice of this group died. The positive control group like the first group 8 days after challenge mortality was started and until the day 18 all mice died. Interesting finding is that the mortality rate was 50% in the third group. These results suggest that the Razi variety of T. gondii caused partially immunity in mice and this variety doesn't cause disease and mortality in mice. The aim of this study was to establish attenuated Neospora caninum (NC-1) tachyzoites through serial passage by J774 cell line. The virulence of attenuated tachyzoites for Balb/c mice were also evaluated. The NC-1 isolates were passage 90 times in J774 cell line to produce high passage or attenuated strain. A total of 35 female BALB/c mice with average age of 6-8 weeks were randomly separated into seven groups. First group as a control was inoculated subcutaneously with cell culture media, the second, third and fourth groups received low passage or acute NC-1 strain with different doses (10 × 10 6 , 5 × 10 6 , 20 × 10 6 ) of tachyzoites. The other three groups inoculated with high passage of NC-1 tachyzoites with similar doses. The mortality rate and pathological changes in all groups were noted daily. The cellular and humoral immunity were assessed by Skin test and ELISA, respectively. The results of the present study showed that mice of the groups that received high passage strain of Nc1 not only survived, but did not show pathological changes of neosporosis. In addition cellular and humoral immune responses to N. caninum were significantly higher in mice that received high passage tachyzoite of NC-1. Background: About 15% of the population in Egypt have chronic hepatitis C and over 90% out of them have HCV genotype 4. The aim of the present study was early predication of the efficacy of HCV treatment with sofosbuvir (sovaldi) and compare between the antiviral efficacy of dual ( plus ribavirin (RBV)) and triple ( plus pegylated interferon (PEG-IFN) and RBV) sovaldi combination therapy. Methods: Blood samples of 100 patients chronically infected with HCV genotype 4 from different Egyptian healthy centers were used for DNA extraction and single nucleotide polymorphism (SNP) genotyping. SNP genotyping for interferon lambda-4 (IFN-L4) gene at ss469415590 was performed by taqMan probe assay. RT-PCR for HCV-RNA was carried out at the start and the end of treatment and twelve and twenty four weeks after the end of the treatment. Hepatitis B virus (HBV) infection is a global concern with an estimated 3.5% of the world population infected. For Malaysia, the national seroprevalence rate of 5% categorises it as a HBV intermediate endemicity country. However, this rate does not represent data from its smallest ethnic minority, indigenous people or Orang Asli. Hence, the objective of this study is to determine the HBV seroprevalence rate among Orang Asli in Malaysia. This is a cross sectional study involving the Negrito, the smallest group out of the three main Orang Asli tribes. This study was conducted in five differently-located Negrito settlements. One hundred and fifty participants were recruited and consented to interview, physical examinations and blood investigation. Sera were tested for Hepatitis B surface antigen (HbsAg) using the Elecsys® HBsAg II assay. Demographic data, biological as well as biochemical parameters were eventually analysed using the SPSS version 20 (SPSS, Chicago, IL, USA). There were 13 participants tested positive with HBsAg. This gives an overall HBV prevalence rate of 14.0% (95% CI: 7.0-21.0%). Male to female ratio was almost 2:1, with eight males and five females infected. Majority (77%) of those infected were more than 30 years old. All of them were born to mothers who were not screened for HBV antenatally. Almost half of them (n = 6) had had past history of tattooing and body piercing. In conclusion, HBV prevalence rate among Orang Asli is almost thrice than that of Malaysia national rate, comparable with studies conducted on indigenous population elsewhere. The possible routes of transmission in this population is either through vertical transmission, as well as practices such as tattooing or body piercing. Background: Dengue case in Malaysia is very common and still on the rise. Due to the right climate variability of high temperature and copious rainfalls, addition to human activity towards urbanization, Malaysia has been classified as endemic country for dengue. The monsoon seasons fall between November to March and from June to September each year. Any patients with acute febrile illness with typical or atypical presentation will be diagnosed as suspected dengue until proven otherwise. Our aim of the study is to determine the correlation between the occurrence of suspected dengue cases with the change of climate in an urban area in West Malaysia. Methodology: A retrospective data collected from 2014 to 2016 at the UiTM Sungai Buloh Clinical Training Centre for all 90 patients with a clinical history of suspected dengue fever were analyzed. The number of suspected dengue fever for each year is tabulated and matched with the distribution of rainfall in West Malaysia. Results: There were peaks of suspected dengue cases in January and July in 2014 and 2015 respectively which coincides with the two monsoon seasons. However, in 2016, the suspected dengue cases peak in March but did not increased during the second monsoon season for that year (Figure 1 ). In 2014 and 2016, there was a period with no suspected dengue cases from February to May and from September to December respectively. This is a period where there is minimum rainfall and drier climate. Conclusion: The finding showed that the peak distribution of suspected dengue cases presented in our centre correlates well with the monsoon season. However, with a period of absence of suspected dengue cases raised a question whether Malaysia is a true endemic country for dengue. Background: Nontypeable Haemophilus influenzae (NTHi) is a significant human respiratory pathogen and a common cause of otitis media, sinusitis, bronchitis and chronic obstructive pulmonary disease. NTHi is capable of forming biofilms, which may be one of the important factors contributing to chronic diseases and accelerating antibiotic resistance. Resistance of bacteria within the biofilms to conventional antimicrobials currently is a major obstacle to effective medical treatment on a global scale. Discovery and development of new agents to effective treatment of NTHi biofilms is therefore necessary and in an urgent need. Methods: In this study, various antimicrobial peptides were synthesized and their anti-biofilm activities against a strong biofilm-forming strain of NTHi were investigated by the microtiter plate biofilm assay. Results: Upon the treatment with the test peptides, modifications on NTHi biofilm formation were observed, but these differed in the degrees. Among the peptides tested, GC2 peptide exhibited the most potent anti-biofilm activity. Its mean minimum biofilm inhibitory concentration (MBIC50) appeared to be 8 μg/mL. Our results indicate that the GC2 peptide has powerful inhibitory activity against NTHi biofilm and warrant further investigations for application of this peptide as a novel therapeutic agent to control NTHi biofilm-associated infections. In vitro pharmacokinetics/pharmacodynamics of the antibacterial moxalactam against extended-spectrum β-lactamase-producing enterobacteriaceae C. Background: The limited choice of drugs for the treatment of infections caused by extended-spectrum β-lactamase (ESBL) producers is a major clinical problem. Objective and Methods: The objective of this study was to evaluate the pharmacodynamics of moxalactam (MOX), cefotaxime (CTX) and cefoperazone/sulbactam (CFZ/SBT) against ESBL-producing E. coli and K. pneumoniae, and non-ESBL-producing E. coli, using an in vitro pharmacokinetics/pharmacodynamics (PK/PD) simulation model. Results: Against non-ESBL-producing E. coli, all dosing regimens exhibited rapid bactericidal efficacy and the regimens achieved 100% of %T > MIC, resulting in a reduction in the bacterial count of >3 log 10 cfu/mL at 24 h. When tested against ESBL-producing E. coli and K. pneumoniae, CTX and CFZ/SBT displayed only weak bactericidal effects, and subsequent regrowth was evident. By contrast, MOX continued to achieve a reduction in the bacterial count of >3 log 10 cfu/mL at 4 h, and MOX (2 g iv q8h) eventually achieved a >3 log 10 cfu/mL reduction at 24 h. And the requirement for %T > MIC for ESBL producers should be higher than for non-ESBL-producing isolates. Conclusion: MOX could serve as an alternative bactericidal agent against ESBL producers. In vitro pharmacokinetics/pharmacodynamics evaluation of fosfomycin combined with amikacin or colistin against KPC2producing Klebsiella pneumoniae W. Yu 1,2 , P. Shen 1 , Z. Objectives: The emergence of carbapenem-resistant Enterobacteriaceae, especially Klebsiella pneumoniae, has become a major concern in clinic settings. Combination therapy is gaining momentum to counter the secondary resistance and potential suboptimal efficacy of monotherapy. The aim of this study was to evaluate the bactericidal effect of fosfomycin (FM), amikacin (AMK) or colistin (COL) alone and combinations against KPC2-producing K. pneumoniae using dynamic model by simulating human pharmacokinetics in vitro. Methods: The Pharmacokinetics Auto Simulation System 400 system was employed to simulate different dosing regimens of FM, AMK and COL alone and combination. Bacterial growth recovery time (RT) and the area between the control growth and antibacterial killing curves (IE) were used as unbiased and comprehensive means for determining the antimicrobial effect. Results: We observed that COL alone was much pronounced than FM or AMK against KPC-Kp. IE of FM (8 g every 8 hours) plus AMK (15 mg/kg once-daily) and FM (8 g every 8 hours) plus COL (75,000 IU/kg every 12 hours) were higher (>170 and >200 LogCFU/mL·h −1 , respectively) than that of monotherapies against sensitive strains. Of note, the rate of resistance was lower when using the combination of FM (8 g every 8 hours) plus COL (75,000 IU/kg every 12 hours) than using COL (75,000 IU/kg every 12 hours) alone. Conclusions: The combination of FM (8 g every 8 hours) plus AMK (15 mg/kg once-daily) and FM (8 g Backgrounds: Although carbapenems remain effective at treating serious multidrug-resistant P. aeruginosa infections, carbapenemresistant P. aeruginosa (CRPA) has now emerged and is disseminating worldwide. Ceftolozane/tazobactam (C/T) demonstrates activity against many multidrug-resistant isolates. We evaluated the activity of C/T and compared its activity to that of ceftazidime/ avibactam (C/A) using a well-characterized collection of CRPA isolates. Methods: Fifty-seven CRPA isolates from a previous study were included. All isolates had been previously evaluated for the presence of bla IMP , bla VIM , bla SPM , bla GIM , bla SIM , and bla KPC by PCR. Expression of oprD, ampC, and several efflux pump genes had also been investigated by qRT-PCR. Antimicrobial susceptibility to C/T as well as C/A was tested by broth microdilution following Clinical and Laboratory Standards Institute (CLSI) guidelines. The CLSI susceptibility breakpoint for C/T of ≤4/ 4 μg/mL and the FDA susceptibility breakpoint for C/A of ≤8/4 μg/ mL were applied. Results: Fifteen isolates, including 13 isolates with bla IMP-6 and 2 with bla VIM-2 showed MICs ≥ 128/4 mg/L for both C/T and C/A. Among 42 carbapenemase non-producing CRPA isolates, overall susceptibility to C/T was 92%, compared to 73.8%, 42.9%, 31.0%, 23.8%, and 21.4% for C/A, ceftazidime, piperacillin/tazobactam, meropenem, and cefepime, respectively. A total of 39 (92.8%) carbapenemase non-producing CRPA isolates showed oprD loss, whilst 29 (69.0%) isolates overexpressed one or more genes of the efflux systems or ampC. Interestingly, C/T MIC values for all mutant strains except three isolates were low. MIC values for C/T ranged from 1 to 32 µg/mL (median, 4 µg/mL) for isolates overexpressing AmpC, whereas isolates with basal AmpC expression levels had MIC values ranging from 1 to 16 µg/mL and a slightly lower median MIC value of 2 µg/mL. Conclusions: C/T was active against a large percentage of noncarbapenemase producing CRPA. Aside from isolates with acquired carbapenemases, no apparent correlation of C/T MIC values to specific mutation-driven resistance mechanisms among CRPA was elucidated. Methods: For covering the gastric mucous layer, two groups of ten rats were administered to either smectite or S-GM. To evaluate anti-H. pylori efficacy, mice were divided into 8 groups, and H. pylori eradication was assessed by Campylobacter-like organism (CLO) test and H. pylori PCR of the gastric mucosa, and H. pylori antigen and PCR of mouse feces. The levels of proinflammatory cytokines were examined. Results: S-GM was retained in the gastric mucosal layer with a >60% distribution ratio for up to 1 h, and the S-GM-based triple regimen decreased bacterial burden in vivo, compared to that in untreated mice or mice treated with other regimens. The therapeutic reactions in the CLO test and H. pylori PCR from gastric mucosa were 70%, 60%, 80%, 50%, 60%, and 60% in Groups III-VIII, respectively. Those of H. pylori PCR in feces of mice were 90% and 100% in Group III and Group V, respectively. S-GM triple therapy also reduced the levels of proinflammatory cytokines. Conclusion: These results suggest that S-GM is a promising and effective therapeutic agent for the treatment of H. pylori infection. Antimicrobial activity of Pseudomonas sp. strain S3.1 against a broad spectrum pathogenic bacteria and fungi S. Magdalena 1 *, Shirley 1 . 1 Faculty of Biotechnology, Atma Jaya Catholic University of Indonesia, Jalan Jenderal Sudirman, Jakarta 12930, Indonesia The use of untreated water for drinking and other activities, like aquaculture, have been associated with infections in humans. The aim of this study was to examine the potential activity of Pseudomonas sp. Strain 3.1 against pathogenic microorganisms and to determine the physiological and biochemical characteristics of the antimicrobial compound. The agar-well diffusion method was used to detect antimicrobial activity. The strain S3.1 of Pseudomonas sp. showed a broad inhibition spectrum with activity against various pathogenic bacteria and fungi. Supernatants showing inhibitory activity were obtained when S3.1 was grown in a range of pH from 7 to 9 and temperatures of 28°C to 40°C. Physico-chemical characterization of the antimicrobial activity showed that it was stable between 4°C and 37°C and its activity was preserved at a wide range of pH. The antagonistic activity observed against the indicator strain possess due to protease and amylase produced by Strain S3.1, suggesting that further studies should be addressed to use Pseudomonas sp. Strain 3.1 as a novel antimicrobial to treat an infectious disease. Multidrug-resistant phenotypes and ESBLs-producing Enterobacteriaceae isolated from animal origins: High alert from Veterinary Hospital! P. Kongsanan 1 *, S. Weerakhun 2 , T. Pradidruengchan 3 , P. Jarupach 3 . The antimicrobial resistance of bacterial pathogens of human and animal origin become a main problem of global public health concern. The use of antimicrobial agents in animal hospital increases a selective pressure and a risk of spread of antibiotic resistant bacteria to human in a community. We determined an occurrence of resistant phenotype from routine bacterial culture in a private veterinary hospital in Bangkok between November 2015 to May 2017. A total of 96 Gram negative bacterial isolates from various types of infections in different species of animals were tested for antimicrobial susceptibility testing. The antibiogram showed that ampicillin (78%) was the most resistant antibiotic followed by norfloxacin (72%), enrofloxacin (63%) and marbofloxacin (50%). The multidrug-resistant (MDR) phenotypes were shown in all isolates of Acinetobacter baummanii, Escherichia coli (62%), Proteus mirabilis (36%), Klebsiella pneumoniae (55%) and Pseudomonas aeruginosa (2%), respectively. Among Enterobacteriaceae isolates, 36% P. mirabilis, 27% K. pneumoniae and 9% E. coli isolates exhibited ESBLs-producing phenotype. The frequency of multidrug-resistant phenotypes and ESBLs-producing phenotype is alarming. Our studies showed the animals in private veterinary hospital as an important reservoir of MDR bacteria. Consequently, the continuous monitoring of antimicrobial susceptibility patterns in veterinary hospitals is required. Furthermore, major dissemination of these resistant bacteria in a community may be unavoidable, if there is no preventive measure in place. This information will be useful to establish an antimicrobial control policy in animal practice to minimize the emergence of MDR bacteria. Background: Respiratory illness, one of the most significant diseases, influences the health and the industry of pigs. Pasteurella multocida and streptococcus suis are the secondary invaders to cause swine respiratory diseases. The aim of this study were to investigate the recent prevalence of P. multocida and S. suis and characterize their antimicrobial susceptibility isolated from lungs of pigs at the slaughterhouses in korea. Methods: A total of 271 lung samples with pathologic lesion were obtained in two slaughterhouses from December 2015 to July 2016. Lung samples were aseptically plated on blood agar and incubated at 37°C in CO 2 incubator for 24-48 h. Presumptive isolates of P. multocida and S. suis were confirmed by PCR. Minimum inhibitory concentrations (MICs) of 40 P. multocida and 56 S. suis against 20 antimicrobial agents were determined according to the CLSI guidelines. Results: A total of 20.7% of P. multocida and 14.8% of S. suis were isolated from lung of pig. In p. multocida, all isolates were susceptible to neomycin, ampicillin, penicillin, spectinomycin, whereas resistant to trimethoprim-sulfamethoxazole, florfenicol, oxytetracycline. Tylosin, sulfadimethoxine and chlortetracycline (97.5%) were the second, followed by tetracycline (95%) and tilmicosin (90.0%). In S. suis, 100% of the isolates showed resistance to tilmicosin. Oxytetracycline (98.2%) was second, followed by enrofloxacin and florfenicol (96.4%), danofloxacin and tetracycline (94.6%), tulathromycin A (89.3%), chlotetracycline and tylosin (87.5%) and erythromycin (85.7%). Conclusion: In this study, the recent prevalence of P. multocida and S. suis were detected. Overall, there were differences in MICs and resistance patterns according to the antimicrobials. The use of antimicrobials in food animals should be monitored regularly to minimize the emergence of resistant bacteria. A case of multiple Abcesses due to Melioidosis: first case reported from North Sumatera, Indonesia A.P. Pasaribu 1 *, S. Pasaribu 1 Melioidosis is an emerging zoonosis which caused by a highly invasive and drug resistant bacterium named Burkholderia pseudomallei. It is found in soil and endemic to Southeast Asia and the Pacific region. B. Pseudomallei exhibits resistance to diverse antibiotics but remain sensitive to some third generation cephalosporins, carbapenems and some fluoroquinolones. In Indonesia, the cases were underreported due to the un-awareness since it can mimic many other diseases. Few cases from Indonesia were reported in Dutch literature and as exported cases in other western countries. A case of 13-year-old boy that was referred to pediatric ward after being treated in surgery ward for 1 week due to recurrent abscesses on his back which was thought as spondylitis tuberculosis. Tuberculosis work-up was negative. The boy used to play football in the mud and swim in the river. The lesions were started as small pustules and then grown in size within 1 month. The abscesses were drainage and treated with a course of levofloxacin unfortunately the lesions persisted. Subfebrile fever was found for 2 months, dry cough in 1 month. He developed pale and decrease body weight in 1 month. Liver and spleen enlargement were found. Pus culture yielded Burkholderia pseudomallei. The patient was treated with ceftazidime and sulfamethoxazole/trimethoprim for 2 weeks and continued with oral sulfamethoxazole/trimethoprim for a period of 3 months. The condition was improved significantly and he has been followed up regularly. It has been reported the first case of Melioidosis in North Sumatera, Indonesia. The patient was treated with ceftazidime and sulfamethoxazole/trimethoprim for 3 months and the symptoms very well improved. Use of plasma exchange followed by convalescent plasma therapy in a critically ill patient with severe fever and thrombocytopenia syndrome-associated encephalopathy S. A 4 year-old boy was presented with fever and cough in July 2016. This patient had been diagnosed with Langerhans cell histiocytosis when he was 20 days old, and had been treated with vinblastine and methotrexate. His blood tests revealed that he had pancytopenia. Blood cultures were positive for Nontyphoidal Salmonella (NTS) serogroup D. All NTS isolates detected in blood (n = 12) and stool (n = 5) specimens, during the 7 episodes of fever or diarrhea observed until May 2017, were identified by the Seoul Research Institute of Public Health and Environment as Salmonella enterica serovar Enteritidis. Initially, the Salmonella infection was treated mainly with ceftriaxone and resolved. The first Salmonella Enteritidis isolate in stool was detected six months after the first detection (January 2017) in blood. The first isolate in July 2016 from blood was susceptible to ceftazidime, cefotaxime, and cotrimoxazole, intermediate to levofloxacin, but resistant to ampicillin. The isolates from blood remained susceptible to 3rd generation cephalosporins, Salmonella isolates started to be detected in stool specimens. The antibiogram of the isolates from blood and those from stool were not identical. The stool isolates were resistant to all drugs tested including the 3rd generation cephalosporin with the exception to levofloxacin. All stool isolates showed synergism using amoxicillin-clavulanate disks and were PCR positive for bla CTX-M , Sequence analysis showed that it is bla CTX-M-79 . The cefotaxime resistance was transferable by conjugation. PFGE of XbaI-digested chromosomal DNA showed all blood isolates have an identical pattern. All four stool isolates had different patterns with 1-3 band difference. In conclusion, repeated isolations of Salmonella Enteritidis with identical PFGE pattern from blood in the immunocompromised patient indicate difficulty in eradication of intracellular Salmonella. As to the Salmonella Enteritidis stool isolates, only 1-3 band difference was noted, suggesting their clonality. The stool isolate with ESBL could have acquired bla CTX-M-79 carrying a plasmid from the intestinal flora. Invasive pulmonary aspergillosis following severe community acquired adenovirus pneumonia H.M. Wong 1 *, Y.Y. Chua 1 . 1 Singapore General Hospital, Singapore Background: Adenovirus is a common pathogen of upper respiratory tract infection and community acquired pneumonia (CAP). Most infections are often self-limiting but severe illnesses do occur in immunocompromised hosts. Rare cases of severe adenovirus pneumonia in immunocompetent adults have been reported with a general poor outcome. Invasive pulmonary aspergillosis (IPA) following adenovirus pneumonia has rarely been described, unlike IPA occurring after influenza infection which is an increasingly recognized entity. Of late, critically ill patients requiring extracorporeal membrane oxygenation (ECMO) is also deemed to be an emerging risk factor for IPA too. Method: Herein we describe a patient with IPA following severe CAP due to adenovirus who required ECMO. Results: A 58-year-old male with diabetes mellitus was admitted for a CAP whose symptoms started 1 week ago. 2 days after admission, it progressed rapidly to acute respiratory distress syndrome (ARDS) while on CAP treatment. He needed inotropic support, mechanical ventilation with ECMO. Polymerase chain reaction testing done on endotracheal aspirate sample was positive for adenovirus. Along with broad spectrum antibiotics, he received 12 days of hydrocortisone and 2 cycles of weekly intravenous cidofovir for severe adenovirus pneumonia related ARDS. ECMO was weaned off 18 days later, but he remained ventilator and inotropic dependent 3 weeks into stopping ECMO. Coupled with the increasing inflammatory markers, a computer topography of the chest was done. It revealed an "air crescent" sign suggestive of IPA which was confirmed on the fungal culture of the pleural fluid on the ipsilateral side. His clinical status continued to deteriorate despite aspergillosis treatment and demised eventually. Conclusion: IPA should be considered in a non-immunocompromised host in the setting of severe adenovirus pneumonia requiring ECMO support which may be potential risk factors. Reports about Trichosporon fungemia among immunocompetent hosts without malignancy or hematologic disease are extremely rare. Reported was an unexpected fatal case of Trichosporon asahii fungemia due to multiple soft tissue abscesses occurring in immunocompetent patients without any known immunocompromising conditions. A 72-year-old man was admitted to hospital due to painful swelling and febrile sensation on both legs lasting for a week. A CT on lower extremities showed numerous tiny fluid collections and diffuse subcutaneous fat infiltration on both whole legs. During the empirical IV unasyn treatment, the clinical course did not improve. Trichosporon asahii was isolated from 3/3 blood and multiple wound cultures, and IV fluconazole started. Laboratory examinations for detecting hidden autoimmune diseases or hematologic diseases were extensively checked, but any Figure: (abstract P2-CC02) Changes in viral RNA load, IP-10, IFN-γ and IgG IFA titer timing of therapies. CSF, cerebrospinal fluid; SFTSV, severe fever with thrombocytopenia syndrome virus significant diagnosis was not defined. In spite of orthopedic management and IV antifungal therapy, his condition had not improved and he died due to uncontrolled severe septic shock on 23 rd hospital day. Because Trichosporon species isolates from blood cultures of immunocompetent patients has been considered as contamination, it has been usually considered as meaningless finding by many physicians. However, Trichosporon species can cause a severe fatal infection in the patients without any immunocompromising conditions like this case, great caution might be taken in some clinical situations. A 7-year old boy was referred to pediatrics outpatient clinic with mild fever for one week, abdominal pain, and ultrasonography (US) finding of hepatic subcapsular abscess. Past medical history revealed that he had acute appendicitis two years ago and perihepatic abscess one year ago. US-guided aspiration and culture of the previous liver abscess grew Entercoccus avium and Bacteroides fragilis. Initial laboratory results at our hospital were following; WBC 9,300 ×10 3 /μL (neutrophil 61%, lymphocyte 21%), hemoglobin 11.3 g/dL, platelet 486 ×10 3 /μL, CRP 4.56 mg/dL (0∼0.3), total bilirubin 0.3 mg/dL, AST 22 U/L, and ALT 7 U/L. Abdominal CT showed a 4cm-sized thick walled, internal septated abscess on the right posterior perihepatic space without hepatic involvement which was thought to be associated with dropped appendicolith. On hospital day 3, US-guided aspiration and percutaneous abscess drainage was done. Bacteroides fragilis was cultured from the pus and Fusobacterium nucleatum, subscepcies vincetii was also identified via bacterial rDNA identification. The patient was treated with IV Piperacillin/tazobactam for 15 days, metronidazole for 7 days, and PO Amoxicillin/clavulanate for the remaining treatment period of 5 weeks. Abdominal US finding before discharge showed collapsed abscess cavity with 1 × 0.5 cm sized residual fluid collection and small appendicolith. Hereby, we report a case of a patient with two episodes of perihepatic abscess that may have been associated with secondary infection of retained appendicolith after laparascopic appendectomy. Case: A 71-year-old woman presented the emergency department with fever for 7 days and bilateral flank pain for 2 days. The laboratory results and abdominopelvic computed tomography finding were compatible with APN (Fig. 1a) . Without prior exposure to antibiotics, ciprofloxacin (400 mg intravenously twice a day) was started and gram-negative bacilli were isolated in the initial blood and urine culture on the next day. The subsequent blood culture showed GNB again despite 3-day ciprofloxacin therapy. After the change in antibiotics to ertapenem (1 g intravenously once a day), the patient's clinical course started to improve. ESBL-producing E. coli isolates were identified in all three consecutive blood samples. PFGE patterns, serotypes, and sequence types showed the three isolates were derived from the identical strain (Table 1 ). The MICs of ciprofloxacin by the broth microdilution method increased from 0.25 to 0.5 mg/L, but remained within the susceptible range. All isolates were resistant to all cephalosporins except ceftazidime while susceptible to carbapenems. The isolates produced CTX-M-14 type ESBL belonging to ST69 clonal group. The failure was attributed to GyrA mutation encoding Ser83Leu within quinolone resistancedetermining regions. Our case suggests that ciprofloxacin should be used cautiously in the treatment of serious infections caused by ESBLproducing E. coli, even in APN, due to ciprofloxacin-susceptible isolates. Six months earlier, he was diagnosed with HIV-1 infection and treated with antiviral agents because of a CD4 T-cell count of 72/ mm 3 . An interferon gamma assay was positive at the time of HIV-1 diagnosis, and a chest X-ray revealed no active lesions. Therefore, he was treated for latent tuberculosis with isoniazid, simultaneously with antiretroviral therapy. However, 1 month after treatment initiation, pulmonary tuberculosis with multiple tuberculous lymph nodes in the supraclavicular and mediastinal areas was detected and confirmed by growth of Mycobacterium tuberculosis cultures from both sputum and lymph node biopsy specimens. As a result, he was treated with isoniazid, rifabutin, ethambutol, and pyrazinamide. After 4 months of treatment, a visible mass was detected in the right supraclavicular area, and his CD4 T-cell count had increased to 521/mm 3 The prevalence of carbapenem-resistant Enterobacteriaceae (CRE) is increasing and treatment options for CRE infections are limited. We describe a case of severe CRE infection successfully treated with fosfomycin monotherapy. Case Description: A 63-year-old Singaporean man with poorly controlled diabetes mellitus (HbA1c 8.1%) was admitted on 12.12.16 to a Saudi Arabian hospital with symptoms of diarrhoea complicated by acute renal failure requiring hemodialysis support. He then flew back to Singapore on the 22.12.16 and was found unresponsive during flight, requiring admission to the intensive care unit. He spiked a fever on the 15 th day of admission (6.1.17), for which blood cultures grew CRE Klebsiella pneumoniae (see Table 1 ). He was treated with combination of intravenous (IV) meropenem and polymyxin B and then IV tigecycline and polymyxin B. After completion of 2 weeks of antibiotics, he developed fever again. Urine and blood cultures both grew Klebsiella pneumoniae. The isolate was found to express both NDM and OXA type carbapenemases. He continued to have persistent fever despite combination therapy with meropenem and polymyxin B and blood cultures were repeated. Subsequent resistance pattern revealed polymyxin resistance for which MCR-1 was negative. He was started on IV cotrimoxazole, but was changed to IV fosfomycin 6 g Q6H in view of worsening renal function. An US of the abdomen showed a left kidney complex cystic lesion. Radiologically-guided drainage of the abscess was done and cultures grew the same isolate. He was successfully treated with 4 weeks of IV fosfomycin. Repeat imaging at end of therapy showed resolution and he was discharged well. Discussion: CRE infections are hard to treat with limited options and optimal treatment is not clearly defined. Here we report on the successful treatment of a CRE renal abscess with drainage and prolonged course of IV fosfomycin. Dengue virus is causative agent of Dengue fever that transmits by Aedes mosquito. There was progressively increasing the geographic distribution since the past several decades especially in tropical regions. This virus infection is self-limited and some of patient is life-threatening illness. However, this viral infection still lack of specific treatment, and the vaccine also not fully effective prevention depend on serotypes of virus. Hence, the secondary infection of this virus is important and must be special considered for vaccination in endemic area. In this study, we investigated the distribution of Dengue virus in Southern part of Thailand in epidemic season by collected the 242 serum samples from Dengue patients and performed the detection by nested RT-PCR, to assess the serotype and find out their relationship with clinical presentations. The results presented 12% presented negative results for RNA detection. The 64% of serum sample were positive for Dengue virus serotype 2, and follow by serotype 3, 1 and 4 as 24%, 8% and 4% respectively. Moreover, the patient with dengue shock syndrome was found in cooperatives with Dengue serotype 2% and 0.44% of patient confirmed with secondary infection with present the clinically as warning sign. We anticipate that results will be provided the information about the serotype circulation and clinical correlation of Dengue virus for a wider starting point discussion and consideration to improve the precise diagnosis and also a preventing strategies planning. Background: Acinetobacter baumannii has emerged as a significant opportunistic pathogen that is involved in severe nosocomial infections. There are also some reports that this bacterium is associated with premature contractions and chorioamnionitis during pregnancy. However, reports associating A. baumannii in causing preterm premature rupture of membrane (PPROM) is limited. In this study, we compared the incidence of A. baumannii in pregnant women with PPROM and those who delivered at term and also, to determine subsequent transmission of the bacterium to their babies. Materials and Methods: Samples consisted of high vaginal swabs from 108 women with PPROM and 110 women with normal pregnancy. For babies (n = 111), swabs were collected from their ears and axillae within six hours of delivery. All the swabs were cultured for A. baumannii on Acinetobacter selective media (CHROMagar) and the suspected colonies were then identified by VITEK ® 2, using ID-GNB card. Results: A. baumannii was isolated from eight (7.54%) mothers with PPROM and eight (7.24%) mothers with normal pregnancy. No significance difference ( p > 0.05) between these two groups were observed. Amongst the PPROM mothers who were A. baumannii colonizers, 50% of them transmitted the bacteria to their babies compared to 25% of those with normal pregnancy. Conclusion: The incidence of A. baumannii colonization in pregnant women is low, regardless whether they have PPROM or not. This suggests that colonization of A. baumannii in pregnant women has no significant role in leading to PPROM and mothers who have PPROM have a high possibility of transmitting the bacterium to their babies. Background: Haemaphysalis longicornis act as the major vector of SFTS virus (SFTSV). Tick bites are the most important risk factor for human infection, transmission of SFTSV from fatal SFTS patient to healthcare workers was possible by blood and respiratory secretion in nosocomial area. Herein, we studied about possible the indolent transmission of SFTSV in patient's asymptomatic family member without tick bite history. Method: We introduced a surveillance of the family members of the SFTS patients from 2016 to June 2017. An epidemiological investigation and blood samples were collected from 9 family members within two weeks after index patient" SFTS diagnosis. Real-time polymerase chain reaction for molecular analysis was using for diagnosis tool. Results: Ten SFTS patients were confirmed in this study period and all were improved. Sixteen family members of 10 SFTS patients were investigated and sampled. Among the total 9 family cluster, two family cluster (4 family members) had positive results of SFTSV. They were no symptom with SFTS and did not have a history of contact with the blood and body fluid to the index patient. However, they had been staying in the same location within prehospital period. We examined to prove the transmission mode in family members for SFTS virus identification using molecular method from loose stool since 2017, SFTSV was negative from four stool specimens in 2017 SFTS patients. Conclusion: To the our knowledge, this is the first report that the results of the study highlighted a potential secondary infection without tick bite, blood contact, or bloody secretion in the SFTSV endemic area. Although, it is difficult to determine the scope of surveillance due to the SFTS transmission mode remain unclear and evidence for the possibility of droplet transmission remain limited till now, we suggest that standard and droplet precautions are required for family member and direct contact for febrile patients in epidemic period. Hydatid cyst alive protoscolices inhibit growth of melanoma tumor on animal model Background: Acute pyelonephritis (APN) has been the leading cause of community-acquired bacterial infection. Material and Methods: Using the claim database of the Health Insurance Review and Assessment Service, the hospitalized patients aged ≥15 with N10 (acute tubulo-interstitial nephritis) and/or N12 (tubulo-interstitial nephritis, not specified as acute or chronic) in ICD-10 codes as primary discharge diagnosis were selected from 2010 to 2014. Results: In total, 208,652 events were included. Geographic analyses revealed a high relative risk (RR) of APN incidence in most areas of Gyeongsangnam, Jeollanam, and Jeollabuk provinces (12.4/10,000 populations, RR 1.41, P < 0.001) and Gangwon province (13.7/10,000 populations, RR 1.44, P < 0.001) (average: 9.6/ 10,000 populations). The average medical costs for treatment of APN was 1,316,000 won and higher in metropolitan areas; Seoul (1,458,000 won, RR 1.13, P < 0.001) and Busan (1, 441, 703 won, RR 1.27 , P < 0.001). Average hospital days was longer in most areas of Jeollanam, Jeollabuk, and Chungcheongnam provinces (9.8 days, RR 1.11, P < 0.001) and Busan and surrounding areas (9.9 days, RR 1.12, P < 0.001) (average: 9.0 days). Higher amount of antibiotic was prescribed for the treatment of APN in Daejeon and surrounding areas (14.0 DDD, RR 1.24, P < 0.001), western areas in Jeollabuk province (12.2 DDD, RR 1.09, P < 0.001) and northern areas in Gyeonggi and Gangwon provinces (11.9 DDD, RR 1.05, P < 0.001) (average: 11.3 DDD). Conclusion: Regional differences in epidemiology of APN were noticed, and the antibiotic usage for treatment of APN was also different by regions in Korea. Methods: Genotyping was performed using the multilocus sequence typing (MLST) scheme. The contigs were queried in the Center for Genomic Epidemiology web-based method for MLST (https://cge.cbs.dtu.dk/services/MLST/) to obtain both allelic profiles and STs. New alleles and allelic profiles were submitted to the online MLST database curator (https://pubmlst.org/cdiphtheriae/). Genetic relatedness between isolates was inferred using the goeBURST algorithm found at http://www.phyloviz.net/. Results: A total of 18 different STs were identified; 12 of the isolates have not been previously described. Three new alleles were also identified in dnaK (2 new alleles) and rpoB (1 new allele) locus. The most common STs were ST453 (n = 7, 22.6%), ST141 (n = 5, 16.1%), ST451 (n = 3, 9.7%) and ST248 (n = 2, 6.5%). The eBURST analysis did not reveal any clonal complexes (CCs) with some clones being single-locus variants of another (ST456-ST462 and ST461-ST460-ST459) and 13 singletons. Comparisons between local STs and reference STs available from the MLST website showed that none of the local isolates cluster within 11 major eBURST groups identified in C. diphtheriae. The MLST results showed heterogeneity among the toxigenic C. diphtheriae isolates in Malaysia. These isolates were also unique as none was identified in the major eBURST groups. (13) Chronically cathererized asymptomatic patient with an unrelated reason for admission e.g. hypertension, constipation 9 (3) Other clear source of fever (immunocompetent and haemodynamically stable patient) 45 (14) Elderly patient with non-specific symptoms/signs, e.g. dizziness, breathlessness 24 (8) Other site of infection without fever, e.g. sacral sore, leg ulcer 22 (7) Objectives: Aeromonas species is an increasing pathogen of bacteremia, especially in the setting of healthcare-associated infection. We investigated the risk factors for mortality in patients with Aeromonas bacteremia. Aeromonas bacteremia between January 2006 and December 2015 at a 2,000-bed tertiary care center in Seoul, Korea, were retrospectively reviewed. The analysis was performed to identify factors associated with 28-day mortality. Results: A total of 138 patients with Aeromonas bacteremia were enrolled for this study. Most common pathogen was A. hydrophila (n = 58, 42%), followed by A. caviae (n = 50, 36.2%) and A. veronii biovar sorbia (n = 19, 13.8%). 93 (67.4%) patients were male and the median age was 64.5 years. The most common source of Aeromonas bacteremia was biliary tract in 74 (53.6%), followed by peritoneum in 10 (7.2%), skin and soft tissue in 5 (3.6%), and liver abscess in 4 (2.9). Primary bacteremia with unknown origin was in 39 (28.3%). The overall all-cause 28-day mortality rate was 29%. On univariate analysis, skin and soft tissue infection (P = 0.025), peritonitis (P = 0.035), healthcare-associated infection (P = 0.029), Charlson comorbidity score (P < 0.001) and the Sequential Organ Failure Assessment (SOFA) score (P < 0.001) were significantly related to the mortality. Results: A total of 140 patients with PM were enrolled. The most common underlying disease was diabetes mellitus (47 cases, 33.6%) and the most common predisposing condition was trauma (21 cases, 15.0%). A total of 111 organisms were isolated from 99 (70.7%) patients, and polymicrobial infections were found in 15.2% (15 of 99). Staphylococci (43 cases) were the most common pathogen, followed by streptococci (23 cases). Enteric gramnegative organisms were identified in 32 patients. Methicillinresistant Staphylococcus aureus (MRSA) was identified from 2.9% (4 of 140) patients (community-acquired cases, 2.0% [2 of 102] vs. health-care-associated cases, 5.3% [2 of 38] ; P = 0.30). The inhospital mortality rate was 8.6%. Old age, intensive care unit admission, and septic shock were significantly associated with inhospital mortality. Conclusions: Underlying conditions and severity were associated with in-hospital mortality. MRSA seems to be an uncommon causative organism of PM in Korea. were enrolled, and clinical symptoms were confirmed. The gene for 56-kDa type-specific antigen (TSA56) was amplified by nested PCR, and the nucleotide sequences of the PCR products were determined. The nucleotide sequences of the TSA56 obtained from scrub typhus patients and retrieved from the GenBank database were analyzed phylogenetically. Results: In PCR results, 247 PCR-positive samples were obtained and 218 samples (88.3%) showed Boryong genotype by DNA sequence analyses. Thirteen samples (5.3%) showed Taguchi genotype; other Korean types such as Je-cheon (5), Yeo-joo (1) and Pa-joo related strain (1) were also identified. Japanese types such as Karp (4), Kawasaki (2), Ikeda (2) and Nishino (1) were also found. The most cases were found in the group of age 70-79 (29.6%). Conclusion: In our results, the most prevalent type of O. tsutsugamushi was Boryong in Korea, 2016; however, unusual type originated from Korean and Japanese strains were also found. Consecutive and broad-ranged national survey for prevalence is needed for the control and prevention of scrub typhus in other endemic region of Korea. Background: Group A streptococcus (GAS) is a common pathogen in pediatric patients and often causes acute pharyngotonsilitis and skin and soft tissue infection. In addition, bacteremia can also occur. Method: This is a single-center, retrospective study. From January 2000 to December 2016, pediatric patients ≤18 years old with GAS bacteremia were studied. Results: Among 249 patients with positive GAS culture results, 19 patients had GAS bacteremia. Eleven (58%) patients were male and median age was 7.4 years (range 0.3-17.4 years). Fourteen (74%) patients had chronic underlying diseases. Five (26%) were immunocompromised patients. Eight (42%) patients had lymphatic or vascular malformation and seven of them had inflammatory signs of the lesion. Three (16%) patients had acute lymphoblastic leukemia and presented with only fever without sore throat or other signs of skin and soft tissue infection. Two (10%) patients had underlying chronic kidney disease. Three (16%) patients developed pneumonia and two of them received ventilator care; two patients were previously healthy children and one patient had chronic bedridden condition with microcephaly on home-ventilator care. The 30-day mortality was 6% (1/19). All GAS isolates were sensitive to penicillin. Fifteen were sensitive to both erythromycin and clindamycin (79%). Conclusion: This study showed various clinical presentations of invasive GAS infections. Although uncommon, GAS should be considered as a potential pathogen that can cause bacteremia and serious clinical course. Background: Porcine respiratory disease complex (PRDC) greatly affects the health and production of pigs and causes significant economic losses in the swine farms. The diseases are commonly associated with bacterial pathogens, and identification of the bacterial pathogens is getting a topic of great interest. In this study, PCR and culture methods were compared to detect five bacterial pathogens mostly associated with PRDC worldwide [Pasteurella multocida (PM), Mycoplasma hyopneumoniae (MH), Streptococcus suis (SS), Actinobacillus pleuropneumoniae (APP), and Haemophilus parasuis (HP)] in lung tissue with pathological lesion. In case of culture method, Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) was used to identify the bacterial species following colony isolation by bacterial culture. Methods: A total of 271 lung samples with pathologic lesion were obtained in two slaughterhouses from December 2015 to July 2016. The tissue was homogenized in 15 mL PBS using a stomacher and genomic DNA was extracted from 1 mL of the homogenized solution after spin down. PCRs were performed with each specific set of primers for a target pathogens using the extracted DNA samples. The homogenized samples were also used to isolate bacteria by streaking onto 10% sheep blood agars followed by overnight culture at 37°C. Each suspected colony of PM, MH, SS, APP, and HP was subjected to MALDI-TOF assay to identify the bacterial species. Results: Of 271 tissues samples processed, PCR detected all the five pathogens with the highest detection rate of MH (n = 141, 52%) followed by SS (14, 5.1%), HP (5, 1.8%), APP (5, 1.8%), and PM (4, 1.4%), respectively. In contrast, culture method detected only SS (29, 10.7%), PM (29, 10.7%), and APP (1, 0.3%). None of the samples were positive for MH and HP by culture method. All the culture positive samples for the pathogens were negative by PCR except 4 samples of SS positive. When combined the results of PCR and culture methods, the overall prevalence of PRDC associated bacterial pathogens in pathological lung tissues was highest for MH (141, 52%) followed by PM (33, 12.2%), SS (29, 10.7%), APP (6, 2.2%), and HP (5, 1.8%), respectively. Conclusions: PRDC is polymicrobial disease caused by infection of various combinations of primary and secondary bacterial pathogens. It is important to investigate the relevant significance of bacterial pathogens in PRDC to control the disease. MH was the most prevalent bacterial pathogen in this study, indicating MH is the most significant bacterial pathogen of PRDC in Korea. PM and SS were the second and third prevalent bacterial pathogens, but APP and HP were rarely associated with PRDC. The two detection methods used in the current study showed clearly different sensitivities to each pathogen. MH and HP were detected only by PCR but most PM and SS were identified by culture method followed by MALDI-TOF assay. Moreover, PCR and culture methods detected each different portion of positive samples, respectively, in this study. Therefore, use of combinational method of PCR and culture might be useful to increase the sensitivity for the detection of PRDC associated bacterial pathogens. The obligate intracellular bacterium Anaplasma phagocytophilum causes human granulocytic anaplasmosis (HGA). This bacterium is a tick-borne pathogen mainly transmitted by Ixodes ticks that infect domestic mammals and humans. Molecular characterization of the MSP2/P44 protein of A. phagocytophilum may determine not only if the bacterium is capable of invading hosts but also whether it generates antigenic variation for the purpose of escaping the host immune response, resulting in various pathologic injuries and serious clinical outcomes. In Korea, HGA patients usually present acute fever, muscle pain and headache. In this study, we confirmed seropositivity and/or seroconversion against A. phagocytophilum infection using indirect immuno-fluorescent antibody assay (IFA) from 4 patients with acute fever. We also amplified, cloned and sequenced the part of A. phagocytophilum msp2/p44 genes detected from the HGA patients and then compared them with the msp2/p44 sequences available in GenBank. The comparative data showed that the 4 msp2/p44 sequences obtained from Korean patients had a high similarity (99.17∼97.92%) with one another whereas were significantly different with sequences detected from humans in USA (NC007797, DQ519522 and JQ599137) and China (KC128828 and KC430393) . This result provides a clue to understand the distinct genetic characterization of A. phagocytophilum originated from Korean patients, which might be relevant to clinical outcomes might have suggest helpful information on them. Further study is needed to understand a mechanism on escaping the host immune response through molecular characterization of the MSP2/P44 protein. Results: Pathogens were identified in 22 (56%) of the 39 blood, liver aspirate, and/or vitreous humor samples. Klebsiella pneumoniae was the most frequent organism (12/39, 33%). Candida was next common (8.3%, 3/39). The most common combined infection was liver abscess (16/39, 41%). And, renal abscess or acute pyelonephritis was in 30.8% (12/39). Two died due to this fulminant infection. Final ocular outcomes were 35.9% (14/39) in NLP, 15.4% (6/39) LP, 15.4% (6/39) HM, 7.7% (3/39) FC, and 25.6% (10/39). K. pneumoniae is a poor prognosis factor for ocular outcome (NLP and LP vs. HM, CF, and 20/200 or better) in the univariate analysis ( p = 0.02). Other factors did not reach to the statistical difference. Tree decision analysis also revealed K. pneumoniae as a node to divide outcome ( p = 0.017). Conclusions: K. pneumoniae is the most frequent pathogen of EE and additionally, seems a poor prognostic factor of ocular outcome. Considering the rapid progression and poor prognosis of EE, early diagnosis and immediate management are vital, especially if K. pneumoniae is identified or suspected. Keywords: endogenous endophthalmitis, Klebsiella pneumoniae, ocular outcome P2-CE22 Non-HACEK gram negative bacillus endocarditis from a developing country Dr R. Madhumitha 1 *, Dr V. Ramasubramanian 1 , Dr P. Senthur Nambi 1 , Dr R. Gopalakrishnan 1 , Dr I. Sathyamurthy 1 . Background: The predominant causative pathogens for bacterial endocarditis in nearly 80% of cases continues to be Gram positive bacteria. The emergence of newer risk factors such as prolonged hospitalization, device insertion and escalating antimicrobial resistance has changed the epidemiology of the disease in the developing world. The incidence of Gram-negative endocarditis in the developed world ranges from 1.3% to 10%, mostly with HACEK (Haemophilus species, Actinobacillus actinomycetemcomitans, Cardiobacterium hominis, Eikenella corrodens and Kingella species) organisms. Objective: This study was conducted at a 600-bed tertiary care hospital in South India to review the changing epidemiology and risk factors of non-HACEK Gram-negative bacillus endocarditis. Method: Patient records from January 2010 to December2015 (6 years) with Infective endocarditis (IE) as per modified Duke criteria were analysed retrospectively. Results: A total of 120 cases of definitive IE were analysed (103 native vale and 17 prosthetic valve). In the 71 (59.1%) patients with positive cultures, 60 (85%) were gram positive pathogen. Streptococcus sp were the commonest bacteria isolated in 15.8%. Gram negative bacteria were seen in 11(9.1%)-9 native valve and 2 prosthetic valve. Escherichia coli (7 patients [63%]) was the commonest pathogen, 80% were Extended spectrum betalactamase producers. In majority of patients urinary tract was the most common portal of entry (9/11) and nosocomial infections, particularly dialysis related central venous catheter infections. Septic embolic complications were seen in 6 (54%). Conclusion: Non -HACEK gram-negative bacilli are relatively rare causes of endocarditis. Epidemiology and risk factors for developing gram-negative endocarditis including multidrug resistant pathogens are emerging. Health care contact is a primary risk factor for acquisition of non-HACEK endocarditis. Pacific region. The aim of present study is to evaluate microbiological difference of urinary tract infection by hypervirulent phenotype compared to liver abscess origin. Methods: The study was performed on 68 K. pneumoniae isolates from patients with community acquired urinary tract infection (n = 34) and liver abscess (n = 34) who were diagnosed at Keimyung university dongsan medical center from January 2014 through July 2016. The hypermucoviscous phenotype was confirmed with string test. Capsular serotypes, rmpA, magA, kfu, allS and iutA were identified using specific primers by polymerase chain reaction. We compared antimicrobial susceptibility and virulence factors according to origin. Results: Hypermucoviscous phenotype and serotype K/K2 were less in urinary tract infection than liver abscess. Among virulence factors, rmpA, magA, allS and iutA were less frequently observed in urinary tract infection. In the subgroup analysis of hypervirulent K. pneumoniae, serotype K1/K2, rmpA, magA and iutA were less observed in urinary tract infection. identified and scores assigned as follows: 1 point to temperature ≥38.5°C, neutrophils ≥10 × 10 9 /L, neutrophils ≥79%, LS of 1, CRP ≥50 to <150 mg/L and Urea ≥8.6 mmol/L; 2 points to neutrophilmonocyte (NM) ratio ≥50 to <100, LS of 2, CRP ≥150 mg/L and PCT ≥0.25 ng/mL; 3 points to NM ratio ≥100 and LS of 3. Sum score of 0-2 has a sensitivity, specificity, negative predictive value and positive predictive value of 97%, 22%, 98% and 12% respectively. P value for goodness-of-fit test for the model is 0.18; area under ROC curve is 80.8% (±0.6%) with optimism of 0.2% (±0.6%). Conclusion: Predictive scores for bacteremia could be used to stratify the risk of bacteremia and safely reduce blood culture taking on admission. Background: Rickettsia and Orientia species are gram negative, strictly intracellular bacilli that multiply within the cytosol endothelial cells. Scrub typhus is a mite-borne disease caused by Orientia tsutsugamushi infection. It occurs frequently during autumn of Korea and the incidence of the disease showed tendency to increase with climate change and outdoor activity. Rickettsia typhi, a typhus group rickettsia, is the etiologic agent of murine or endemic typhus. R. typhi is transmitted to human primarily by the rat flea, Xenopsylla cheopis, although lice and mites are also potential vectors. In Korea, about 8,000 cases of Scrub typhus have been reported since the first case reported in the 1950s. However, Murine typhus has been reported about 30 cases. Through surveillance study, we obtained basic data of Scrub typhus and Murine typhus for prevention and control in highrisk population. Materials and Methods: This study was performed for national park workers considered as a high-risk population of Rickettsia disease. The eligible participants were 763 (36.9%) out of 2,067 forestry workers. The sera were collected and then the antibodies specific to O. tsutsugamushi and R. typhi were titrated using indirect immunofluorescence antibody assay (IFA). 1:256 in IgG or 1:16 in IgM was considered as a single cut-off titer for seroprevalence. Results: The seroprevalence rate was indicated as higher titer than the single cut-off and seroreactivity rate was indicated any titration to the single cut-off. Scrub typhus was showed 4.8% (37/763) and 10.0% (76/763), respectively. And Murine typhus was showed 3.0% (23/763) and 8.9% (68/763) respectively. Moreover, seroprevalence of Scrub typhus and Murine typhus was 0.5% (4/763). Conclusions: This is the first serological study of murine typhus among forestry workers considered as high-risk population. The high exposure to R. typhi amongst forestry workers suggests that flea-borne spotted fever is an important cause of undifferentiated fever conditions that may not be adequately recognized in Korea. Therefore, continuous surveillance needs to understand the status and to improve control and prevention of these diseases in highrisk population. We explored the potential effect of gender on clinical outcomes in Staphylococcus aureus bacteremia (SAB HIV prevalence in Malaysia remains high above 5% especially among the high risk populations such as person who inject drug (PWID), female sex worker (FSW), transgender (TG) and men who have sex with men (MSM). HIV infection can be asymptomatic for a significant period of time and this can lead to delayed diagnosis especially among the low risk population. As asymptomatic people are capable of transmitting the infetion, these asymptomatic, low risk population is vital in playing a role to curb the uprising of HIV infection. This study aim to provide preliminary data on HIV prevalence among perceived low risk patient in a specialist center in Malaysia. A retrospective study was done where data of 6,963 patients that were screened for HIV antibody/antigen by chemiluminescence imunoassay (CLIA) during the year 2013 until 2016 were examined. These patients undergo HIV screening as a routine either before cardiac procedure or as part of antenatal checkup. There were no high risk behaviour recorded. Those reactive by CLIA were then confirmed by particle agglutination test. There were 22 (0.3%) new cases detected during the screning. Ninety six percent of them were male. Majority of them (90%) in the age range of 35-60 year. This study showed that screening detects small but significant percentage of new HIV cases among perceived low risk patients and this provides an oppoturnity for early diagnosis and management. The varied rates of immune recovery of HIV patients on HAART by age category and their implications N. Background: Age is a known negative predictor for immune recovery following highly active antiretroviral therapy (HAART). The non-linear association has not been well-characterised. Methods: We collected longitudinal clinical data (1985-2014) from a HIV specialist clinic in Hong Kong. We used CD4, CD8 and CD4/ CD8 ratio to examine the immune recovery of Chinese patients who had been on HAART for ≥4 years. Three age categories at HAART initiation were defined: young adults (18-49), middle-aged (50-64) and elderly (≥65). Late HAART initiation was defined as pre-HAART CD4 ≤100/μL or AIDS diagnosis. Generalized estimating equations were used to analyse the association of age category with the temporal change of clinical immune outcome markers from month −2 to 60. Results: A total of 913 patients (87% male) were analysed. At HAART initiation, 81% were young adults, 14% middle-aged and 5% elderly. The baseline median CD4, CD8 and CD4/CD8 ratio were 128/μL (IQR = 36-204/μL), 704/μL (IQR = 447-1,038/μL) and 0.15 (IQR = 0.07-0.25) respectively. Middle-aged were more likely be in late HAART initiation than young adults (OR = 1.55), but were similar to elderly (OR = 1.38). Performance of CD4 recovery worsened along age categories with the best in young adults (5.26/μL/month), and a reverse for CD8 decline with the largest drop among elderly (−4.34/μL/month) but smallest in young adults (−0.5/μL/month). A U-shaped relationship between age category and the rate of CD4/CD8 ratio recovery was noted with the middleaged having the slowest recovery. Conclusions: While CD4 recovery of middle-aged might occur faster than the elderly, their CD4/CD8 recovery was slower. Immune recovery of middle-aged may improve if HAART could start early so as to shorten the interval from diagnosis to treatment initiation. patients with diagnosis "HIV infection" or "AIDS" were reviewed at a university hospital located at Seoul, Republic of Korea. Acute HIV infection was defined as a patient who had initial screening HIV-1, 2 antibody ELISA negative with subsequent seroconversion within 6 months with any clinical symptom. Results: All 23 patients diagnosed with Acute HIV infection were male and the median age at the time of diagnosis was 33 . The median value of Log 10 HIV viral load by RNA RT-PCT at the time of diagnosis was 5.6 and the median CD4 count was 505/μL. Patients with acute HIV infection most commonly had fever (100%) and (mean duration 8.5 days), followed by gastrointestinal symptoms (83%, nausea and vomiting 52%, diarrhea 44%) and tonsillar enlargement (48%). Leukopenia (WBC count <4,000/μL) and thrombocytopenia (Platelet count < 150,000/μL) were present 74% and 70%. Methods: Reviewed retrospectively were the medical records of 388 HIV-infected patients, who had been treated with HAART in a single tertiary hospital since 1 January, 2012. Hyperlactatemia was defined as serum lactate >2.0 mmol/L. Factors were analyzed through comparison between the group showing increment of serum LA level (group 1) and the control group that showed normal or decreased serum LA level (group 2). Results: Among 189 cases were included, 70 cases (group 1) showed increase of LA during HAART (range 2.0-5.1 mmol/L). There was no difference in the distribution of HAART regimen between two groups ( p = 0.338), and according to the NRTI used in HAART between group 1 and group 2 ( p = 0.446). There was no difference in underlying diseases such as co-infections, metabolic diseases and psychiatric diseases between two groups. However, initial serum albumin (4.7 g/dL vs. 4.6 g/dL in group 1 vs. group 2, p = 0.020) and hemoglobin level (14.8 g/dL vs. 14.3 g/dL in group 1 vs. group 2, p = 0.012) showed a statistically significant difference. Increment of CD4+ T cell count ( Background: Advanced human immunodeficiency virus (HIV) infection can have copious opportunistic infections and HIV related cancers, the relative risk is much higher compared with the general population. Diffuse large B-cell lymphoma (DLBCL) is one of those diseases, it can involve both lymph nodes and extranodal site, but lung involvement is very unusual. Case report: We report a case of pulmonary DLBCL combined with multiple infections in a 43-year-old HIV positive man, presenting with bizarre cystic radiologic findings, whose main symptoms were general weakness and 10 kg weight loss for two months with chronic cough. He was diagnosed with advanced HIV infection, with a CD4 count of 6. The chest radiologic findings identified multifocal septated cystic lesions with thin wall in both upper lungs adjacent to normal parenchyme. The lesions were likely to be in varying states of cystic and necrotic change. As the chest radiologic findings were atypical and clinical signs were unclear, video assisted thoracostomy (VATS) biopsy was conducted for the sake of prompt diagnosis. Its histopathologic finding demonstrated both DLBCL with positive EBV infection and pneumocystis jirovercii (PCP) infection. Mycobacterium avium was also isolated in the AFB culture test. Although chemotherapy could not be performed because of his poor general condition, antiretroviral therapy was started successfully after a few weeks treatment of anti-mycobacterial antibiotics and PCP. His general condition became better and his infectious diseases are under control now. Conclusion: This is the case of pulmonary DLBCL with multiple infections which showed atypical chest radiologic patterns in an advanced HIV patient. Early VATS biopsy was also helpful to have early diagnosis. Background: In May 2015, the 68th World Health Assembly adopted the Global Action Plan on Antimicrobial Resistance (AMR), which reflected the global consensus that AMR threatens human health. Accordingly, the WHO has developed the Global Antimicrobial Resistance Surveillance System (GLASS) to promote standardized AMR surveillance globally. The goal of GLASS is to enable standardized, comparable and validated data on AMR to be collected, analyzed and shared with countries. Since May 2016, the Korea Centers for Disease Control and Prevention has established the AMR surveillance system for general hospitals in six large cities and provinces in Korea, which is compatible with GLASS of WHO and named it Kor-GLASS. From the defined specimens (blood, urine, feces, urethral and cervical swabs), the priority pathogens are isolated, and the antimicrobial susceptibility of all the priority pathogens is determined. Methods: A total of 11 pathogens were tested, including Escherichia coli, Klebsiella pneumoniae, Acinetobacter spp., Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium, Streptococcus pneunoniae, Salmonella spp., Shigella spp., Neisseria gonorrhoeae (8 GLASS-recommended pathogens and 3 pathogens designated as statutory infectious diseases). Antibiotic susceptibility test was performed by disk diffusion and E-test. Both broth microdilution and agar dilution method were used according to the species and antibiotics. The results were confirmed according to Clinical and Laboratory Standards Institute (CLSI) or The European Committee on Antimicrobial Susceptibility Testing (EUCAST). Results: Total 11,093 strains were collected from May 2016 to April 2017 (1 year). Shigella spp. and N. gonorrhoeae were not collected. The resistant rates of E. coli and K. pneumoniae to cefotaxime were 32.1% and 34.5%, respectively. P. aeruginosa and A. baumannii showed 12.6% and 72% resistant to carbapenem antibiotics, respectively. The rates of MRSA in S. aureus isolated form blood was 54%. In Enterococci isolated from blood, the resistance rates to ampicillin were 0.6% and 90.3% for E. faecalis and E. faecium, respectively. S. aureus, E. coli, A. baumannii and E. faecium had more than 50% multidrug resistance. Conclusion: Kor-GLASS may produce meaningful data at a global level to enable analysis of the occurrence and trends of AMR, and establish prevention and control programmes against AMR. In Background and Objectives: Leishmaniasis is a major health problem global and affects millions of people especially in developing countries. Science, there is no immunoprophylaxis (vaccination) accessible for Leishmania infections and commercial drugs are unsatisfactory. Therefore, the objective of the present survey was to state the antileishmanial activity of two herbal medicine (Satureja khuzestanica leaf and Heracleum persicum fruit) extracts were evaluated against Leishmania major and Leishmania infantum using colorimetric MTT (2-(4,5-dimethyl-2-thiazolyl)-3,5-diphenyl-2H-tetrazolium bromide) assay and compared to the Glucantime as a reference. Materials and Methods: The leaves extracts of selected plants were obtained by maceration. The in vitro assays were carried out on Leishmania major and Leishmania infantum using colorimetric MTT assay in comparison with Glucantime. The concentration-response curves tested extracts and glucantime solutions were designed and IC 50 values were located. Results: Anti-Leishmina effects of Satureja khuzestanica and Heracleum persicum on L. major and L. infantum promastigote were revealed with 50% inhibitory concentration (IC50) values of 4.8 and 7.5 mg mL −1 for Satureja khuzestanica, 29.3 and 14.7 mg mL −1 for Heracleum persicum. In the comparison with the standard drug, glucantime, which had IC50 value of 40.2 mg mL −1 for L. major and 18.5 mg mL −1 for L. infantum promastigote after 72 hours incubation, respectively. Conclusions: These results revealed that compounds from Satureja khuzestanica and Heracleum persicum have anti-leishmania properties that necessary to survey the effects of these extracts on leishmania genus in animal models in future. Keywords: antileishmanial activity, Leishmania major, Leishmania infantum, glucantime, promastigote, MTT assay P2-OT03 Identification of pathogenic bacteria on cosmetic testing in public shops in North Jakarta, Indonesia S. Aisyah 1 *, G. Cosmetic testing is one of the essential activities that costumers do before deciding to purchase the beauty product. However, cosmetics samples are used by many people and predominantly have direct contact to open air in which it has the risk of being contaminated by wide range of microorganism. This study aimed to detect the presence of pathogenic bacteria on liquid and solid lipstick in public shops in North Jakarta, Indonesia. Six samples for each lipstick types were randomly selected and cultured in the selective and non-selective medium. Biochemical test were conducted on colonies with different morphology. Of all the samples that were tested, Shigella was found in the 83.3% and 50% in the liquid and solid form of lipstick respectively. On the other hand, E. coli was found in the 66.7% in the liquid lipstick only. Such contamination might come from the users which amenable to the personal hygiene practice and constitute potential hazard to the GI tract of the tester when it accidentally ingested. Further study regarding the ability of the pathogenic bacteria that are exist in the cosmetic sample to transmit disease among the tester is needed. P2-OT04 Implementation of inter-sectorial "One Health" antimicrobial resistance (AMR) containment programme in Malaysia Background: The incidence of AMR is increasing around the world and is affecting all levels of society, thus making AMR a global health threat. AMR is a complex epidemiological problem due to the fact that various causal factors are intertwined. Methods: The Ministry of Health (MOH) Malaysia has taken steps to combat AMR by initiating measures within MOH as well as intersectorally through the One Health approach. In 2016, the MOH steered the initiatives of One Health approach to combat AMR involving various sectors from both human and animal health. This started with the drafting of the Malaysia Action Plan on AMR, focusing on four priority areas which consist of education and awareness; surveillance and research; infection prevention and control; and appropriate use of antimicrobials. At the same time, the National AMR Committee (NARC) was established as a coordinating body to govern the containment activities and its implementation. Under the NARC framework, there are four technical working groups responsible for implementing the planned strategies. Results: Malaysia has successfully established an inter-sectorial One Health AMR framework which governs containment activities as set in the Malaysia Action Plan on AMR. This steering committee is supported by four technical working groups responsible for executing the strategies under their respective priority areas. One of the main outcomes is the implementation of the Integrated One Health AMR Surveillance. Conclusion: AMR is becoming an increasingly urgent challenge and a public health threat for Malaysia. Inter-sectorial collaboration is needed to share evidence on the trends of AMR and the use of antimicrobials in all the related sectors. Awareness and education, infection prevention and control and the antimicrobial stewardship programme will be the driving force to contain AMR. The success of these measures can only be achieved through One Health approach. Background: Cancer is the main cause of death in the developed countries. There are some scientific evidences indicating that parasitic infections induce antitumor activity against certain types of cancers. Hydatid cyst is the larval stage of Echinococcus granulosus, that causes hydatidosis in human and livestock. In this study, the effects of alive hydatid cyst protoscolices on Melanoma tumor in C57/black mice was investigated. Materials and Methods: In this investigation two groups of mice were challenged with B16F10 Melanoma cells. After 2 weeks one group was injected with 500 alive protoscolices as case group and the next group left intact as control group. Tumor size in the case and control groups was measured and the data were analyzed using SPSS software and one-way Anova test. Result: our results demonstrated that alive protoscolices of hydatid cyst inhibit melanoma tumor growth in the case group and difference between mean size of tumor in case and control group was statistically significant. Conclusion: The results of this study showed that injection of alive protoscolices of hydatid cyst had significant effect on melanoma tumor growth. Backgrounds: Deep-seawater (DSW) contains several minerals beneficial for human bodies. The effect of DSW has been known to improve specific immune functions. The beneficial effects of DSW suggest that it could be used for development of new health functional foods. The purpose of this study was to evaluate microbiological food safety of meat products processed with DSW. Materials and Methods: Concentrated DSW (CDSW) containing 5.6% salt was used. Bacterial killing activity was measured against E. coli, Salmonella typhimurium DT104, and S. enteritis, respectively, by incubating the bacteria in CDSW or 0.85% saline (control) at 4°C, and measuring viable bacteria after 24 and 72 h. Two different pork sausages were produced using traditional curing solution or CDSW. Bacterial contamination in the sausages was measured at different time points. Result: The bacterial survival in CDSW and control saline was assessed by measuring colony forming unit (CFU) at different time points. The survival rates of three tested bacteria were more rapidly decreased in CDSW than in control solution. In the bacterial contamination study, no bacteria was detected in either of traditional or CDSW sausages at day 0 and 10. Conclusion: DSW showed some degree of bacterial killing activity, and pork sausages made with DSW showed no bacterial contamination and kept sterile condition until 10 day post production. The contamination study is now in progress to extend the time points being measured. The result indicates DSW could be used for an alternative curing solution in processing meat products. Background: Sitafloxacin exhibited a broad antibacterial spectrum and excellent antimicrobial activity against common bacterial pathogens causing different infections. Sitafloxacin has also exhibited excellent clinical efficacy in patients who had failed to respond to other antibacterial therapy. However, sitafloxacin has not been available in Vietnam yet, so that the local information of the in-vitro spectrum of this antibiotic to the most common bacterial pathogens isolated from Vietnam is very necessary to help the doctor having more choice to against the existing situation of antibiotic resistance in Viet Nam. Main aims of the study: Using the microdilution method to determine ratio of sensitivity and the MIC 90 of Sitafloxacin and other common used antibiotics against the most common bacterial pathogens isolated from the clinical samples collected from patients with acute lower respiratory tract infections and with acute urinary tract infections. Objects and methods: The objectives of the study are the bacterial pathogens isolated from sputum including S. pneumoniae, H. influenzae, M. catarrhalis, S. aureus, K. pneumoniae, P. aeruginosa, and A. baumannii; and from the urine including E. coli, P. mirabilis, E. faecalis, E. faecium, A. baumannii, P. aeruginosa, and K. pneumoniae. For each pathogen, at least 30 isolates are collected. The isolates are collected from different hospitals in Ho Chi Minh City and in Ha Noi for one year (2015). The determination of the MIC of the antibiotics against the bacterial pathogens were carried out by the broth microdilution method following CLSI 2015. To carry-out the MIC, the plates with 96 wells containing the dry antibiotics with serial concentration custom fabricated and supplied by Eiken Chemical Co., Ltd. (Dry Plate Eiken, Tokyo, Japan) were used. Results and discussion: From 1/2015 to 12/2015, 682 isolates were collected from 4 hospitals in Ho Chi Minh and 1 hospital in Ha Noi including 457 isolates from lower respiratory tract infection and 225 isolates from urinary tract. The received results demonstrated that 100% of H. influenzae were sensitive to sitafloxacin, equivalent to amoxicillin/clavulanic acid, cefotaxime, ceftriaxone, and imipenem, however sitafloxacin gave the very low MIC 90 , only 0.12 µg/ ml, the lowest versus other antibiotics. Against S. pneumoniae, 100% of the isolates were sensitive to sitafloxacin and the MIC 90 was 0.06 µg/ml, the lowest versus other antibiotics. Sitafloxacin was among the antibiotics with very high sensitivity ratio to M. catarrhalis (96.2%) and gave the lowest MIC 90 0.12 µg/ml. Against the cocci Gram [+] like MRSA, MSSA, E. faecalis and E. faecium, sitafloxacin gave the very high sensitivity ratio, equivalent to vancomycin (MSSA) or only after vancomycin (MRSA, E. faecalis, E. faecium), however in most of the cases, higher than the sensitivity ratio of the other antibiotics. Against A. baumannii, sitafloxacin gave the sensitivity ratio 97.6%, over all other antibiotics that its sensitivity ratio was not over 25%. Against P. aeruginosa, sitafloxacin gave the same results, the sensitivity ratio (80.7%) were highest compare to other antibiotics. Against ESBL [+] E. coli, sitafloxacin gave the sensitivity ratio 95.1%, only after meropenem (100%), and over all other antibiotics. Against ESBL [+] K. pneumoniae, although the sensitivity ratio could reach only 46.9%, however this ratio was only lower than meropenem (59.4%) but higher that other antibiotics (most gave the sensitivity ratio not over than 25%). Against E. coli and K. pneumoniae ESBL [-] , as well as Proteus spp., sitafloxacin was among the antibiotics with the highest sensitivity ratio. Conclusions: This is the first trial to find-out the in-vitro activity of Sitafloxacin against the different bacterial pathogens isolated from clinical samples in Viet Nam. The results collected from the study demonstrated that Sitafloxacin exhibited excellent in-vitro antibacterial activity against both Gram-positive and Gram-negative bacteria that are mostly encountered in the clinical fields of infections. This result can confirm that Sitafloxacin can be considered as one of the most powerful antibiotics that doctor can use to treat various infections caused by antibiotic resistant bacterial pathogens. This retrospective study aimed to evaluate the levels of coagulation factors and presence of disseminated intravascular coagulation (DIC) in patients with scrub typhus. We included patients confirmed to have scrub typhus at the Chosun University Hospital between September 2004 and December 2009. The DIC scores were evaluated in 365 patients and 36 healthy controls. The median concentrations of fibrinogen, D-dimer, and fibrin/ fibrinogen degradation products (FDP) were compared between patients and healthy controls ( p < 0.001 for all tests). Patients with scrub typhus had longer prothrombin time and lower platelet counts than the controls. Major bleeding was observed in 18/365 patients with scrub typhus. Fifty-one (14.0%) patients presented with severe complications of scrub typhus. Overt DIC and thrombocytopenia (<100,000 platelets/mm 3 ) were observed more frequently in patients with bleeding and severe illness. Furthermore, median platelet counts were low in both groups. Approximately 2.7% (n = 10) and 16.4% (n = 60) patients with scrub typhus had overt DIC, as defined by the International Society on Thrombosis and Hemostasis DIC score (DIC1) and the DIC-scoring template with a fibrinogen/C-reactive protein-ratio (DIC2), respectively. Three (16.7%) and 10 (55.6%) patients with bleeding had overt DIC, as defined by the DIC1 and DIC2, respectively. Seven (13.7%) and 26 (51%) patients with severe illness had overt DIC, as defined by DIC1 and DIC2, respectively. In conclusion, activation of the coagulation system is an important feature of scrub typhus and is correlated with severe disease, including bleeding. This is the first study to report a relationship between DIC and scrub typhus. Background: Quality manuscript editing is necessary for publication in international medical journals, especially for non-native English-speakers. External editing services (EES) are popular, but authors are often unsatisfied with the cost, timeliness, and quality of the services. As a result, our institution launched an in-house editing service (IHES) that works closely with authors to more personally meet their needs. Methods: We designed a survey to monitor author satisfaction and compared preliminary data from one survey item to corresponding results for EES used by our institution in the past year. Authors rated our IHES using a 5-point Likert scale to answer the following questions: (1) Was the request finished in a timely manner? (2) Did the editor fully comprehend the manuscript? (3) Did the corrections correspond with your intentions? (4) How many additional corrections did you make? (5) Was 1:1 consulting helpful? (6) How would you rate your overall satisfaction with this service? (7) Was this your first time using this team's service? (8) Was this your first time using any editing service? (9) How would you rate your relative satisfactory rate compared to other services? (10) Would you use this service again? (11) Would you recommend this service to your peers? Results: The overall satisfactory rate for our IHES was 4.72±0.50 (mean±SD). For question #2, our IHES scored 4.80±0.54, which was significantly higher than corresponding results from another survey evaluating 6 EES (mean, 3.98±SD; range of means, 3.74-4.60; p < 0.001). Conclusion: Authors were highly satisfied with our newlyestablished IHES, and they deemed our editors as having fuller comprehension of their manuscripts than did the EES editors. Background: In Singapore, there are currently two routes to pursue a career as a Medical Technologist (MT) via Bachelor Degree in Biomedical Science or its equivalent from two local universities or Science-related Diploma/Special Diploma from any the five Polytechnics. Academic training covers the core disciplines in laboratory medicine (e.g. Haematology, Clinical Chemistry, Medical Microbiology, Transfusion Medicine and Cyto-Histopathology). Students passed theoretical examinations and completed practical placement in a working laboratory are deemed competent as MT. This report summarises the academic training of the Microbiology MT. Method: Typically Microbiology discipline involves many manual processes from culture plating to recognising, identifying potential pathogen and their susceptibility to antibiotic. Today bench Microbiology MT needs to acquire skills and knowledge on automation; walk-away sample plating and automated identification and susceptibility testing system. Learn newer mass spectrometry technique that allows bacteria identification with reduction in turnaround time (TAT) to minutes. Acquire molecular technique to improve TAT, sensitivity and specificity. Develop medical informatics and process algorithm for management, process control, decision making on scientific analysis, expert reporting of antibiotics, antibiotic stewardship support and accurate reflex testing. Emphasis in quality management and quality control plan are essential. Continuous skills and knowledge improvement to maintain competency and currency in Microbiology is crucial. Result: Current academic programs need to provide the necessary professional development for relevancy of dynamic changes in today's laboratory services. It has been long-debated whether multiple sputum specimens, three to be exact, are absolutely required for pulmonary tuberculosis (TB) diagnosis. Studies from high TB burden country have shown that it can be diagnosed effectively using only two specimens. However, researchers in low TB burden countries have presented contradictory findings. This study was set out to evaluate the optimum number of sputa required to diagnose TB by acid fast bacilli (AFB) smear microscopy, in a moderate TB burden country such as Malaysia. We retrospectively reviewed acid-fast bacillus (AFB) smear Kinyoun stain microscopy result of three consecutive sputum specimens from 31 culture-confirmed TB patients. This cohort of patients includes all inpatients and outpatients presented from 2011 to 2015. HIV patients and foreigners originally from high, or low, TB burden countries were excluded. The data of 31 AFB positive patients were eventually analysed. Twenty-six of those (83%) were successfully diagnosed based on their first sputum. Twenty-nine patients (94%) were AFB positive based either on first sputum only, second sputum only or both. This meant that collecting two consecutive specimens improves AFB detection by about 10%. Finally, two patients (6%) required third sputum examination to eventually clinch their diagnosis. Overall, the third sputum did not appear to have significant additional diagnostic value for AFB detection by microscopy. We concluded that examining only two sputum specimens is sufficient for an effective diagnosis in a moderate TB burden country. It also lessen the financial implication to both patient and healthcare system, to the latter by decreasing laboratory workload and shortening the period of mandatory TB isolation for smear-negative patient. Background: Rapid differentiation of the Mycobacterium tuberculosis complex (MTBC) and Nontuberculous mycobacteria (NTM) is important for early and effective treatment. MPT64 is a MTBC specific antigen secreted during bacterial growth, which has been shown to be an excellent target for differentiating MTBC from NTM. In this study, we investigated the frequency of MPT64 Ag negative and IS6110 PCR positive MTBC strains isolated from clinical specimens in Korea. Methods: January 2014 to December 2016, AFB cultures were performed on clinical specimens using 2% Ogawa medium and BACTEC MGIT 960 System (BD Diagnostics, USA). AFB staining and MPT64 Ag assay were performed on cultured strains. For MPT64 Ag negative strains, IS6110 PCR was performed. The first strain of the multiple isolates from the same patient was included in this study. Results: Of the 5,749 AFB culture positive specimens, MPT64 positive strains were 3,586. Of the 2,163 MPT64 negative strains, 129 (6.0%) were IS6110 PCR positive strains. Of the 3,669 cases reported in MTBC, 92 cases (2.5%) was MPT64 negative strains and among the 46 cases reported in mixed growth (MTBC and NTM), 37 cases (80.4%) was MPT64 negative. Conclusions: The prevalence of MPT64 Ag negative and IS6110 PCR positive MTBC strains isolated from the clinical samples in the last 3 years in Korea were 3.5% (129/3,715) . Of MTBC strains mixed growth with NTM, 80.4% (37/46) was MPT64 negative. The false negativity may be attributed to the lower concentration of MPT64 protein and could also be explained by the deletion and mutation in the mpt64 gene. In this study, mpt64 gene PCR and sequencing was not performed. A negative MPT64 assay cannot exclude the diagnosis of tuberculosis. For MPT64 negative isolates, conventional or molecular tests are required to confirm the presence of MTBC. Neonatal splenic and hepatic tuberculosis: case report and review of the literature Z. Yang 1 . 1 Beijing Children Hospital, China (P.R.C) Background: Congenital tuberculosis is rare, even where tuberculosis (TB) is endemic. A 13-day-old girl presented with a 6-day history of fever, originally the infant abdominal ultrasound showed multiple nodes in liver and spleen after 3weeks antibiotic therapy, including using linezolid in venous for 2 weeks the nodes decreased but did not disminished then the linezolid was changed to orallyadministrated sequential therapy, but fever recurrence and neurological symptoms appeared after discharging for 1week. Her mother was asymptomatic, and her chest radiograph didn't show any evidence of TB infection like bilateral miliary nodules. PPD test was negative. But finally diagnosed with a TB infection by a positive T-SPOT. Congenital TB was strongly suspected because of the symptoms, signs and maternal TB history, and was confirmed by T-SPOT. Timely administration of standard anti-TB therapy combined linezolid resulted in a good outcome of the patient. The case highlights the importance of differential diagnoses of splenic and hepatic tuberculosis and linezolid for neonatal TB may facilitates treatment and the prognosis,but cannot be single used. Case Presentation: We reported a complicated case of congenital TB. A 13-day-old girl presented to the Department of Pediatrics" Emergency Room, Beijing Children's Hospital (BCH), Linkou with fever of 6 days" duration. She was the second child of a 22-year-old woman and had been born in local hospital by Caesarean section at 37 weeks gestation weighing 2,600 g. Her temperature was 37.2°C and Apgar scores were 8 and 9 at 1 and 5 min, respectively. On admission, she weighed 2,380 g and her temperature was 38.8°C, pulse 166 beats/minute, blood pressure 82/64 mm Hg and respiratory rate 38 breaths/minute. Chest wall retraction and coarse breath sounds were present. The remainder of the physical examination was normal. There was no cyanosis, lymphadenopathy or hepatosplenomegaly. Previously, the infant's mother had been healthy with no history of TB contact. Conclusion: Through this case, we aimed to emphasize the importance of including Congenital tuberculosis in the differential diagnosis when a young infant admit with fever no reason and abdominal ultrasound show disseminated hepatic and Splenic nodal, and linezolid can be useful in treatment of Congenital tuberculosis. but it can not be single used and its course is on controversy. makes early recognition crucial and life-saving. Results of TB patients seen at Faculty of Medicine UiTM in between January 2011 to December 2016 were retrospectively analyzed. All culture and sensitivity test results performed using BD BACTEC MGIT 960 SIRE system was included. Culture results without comprehensive susceptibility pattern were excluded. First line drugs included for the data were streptomycin (STR), isoniazid (INH), rifampicin (RIF) and ethambutol (ETH). A total of sixty-seven positive results for MTB culture and sensitivity were obtained. Sixty-two isolates (92.5%) were sensitive to all four antimycobacterials tested. Four isolates (6%) demonstrated mono-resistant susceptibility pattern; two were RIF-monoresistant, one were INHmonoresistant and the other one, ETH-monoresistant. One isolate (1.5%) were found to be resistant against two drugs, isoniazid and streptomycin. Over 90% of Mycobacterium tuberculosis isolated in this centre was sensitive to four first line drugs tested. Although five isolates were resistant to either one or two antimycobacterials, none of those were resistant to both isoniazid and rifampicin. This meant that MDR-TB was not present in this particular cohort of patients. Still, efforts should be made to ensure that these drugsusceptible patients do not develop MDR-TB in future. Comparison of results is difficult while standardisation of results is a challenge. Methods: We examined the performance of 3 different loci sets for genotyping 52 MTB isolates collected over a 2-year period from a large regional hospital in Hong Kong, an intermediate TB-burden area. Results were compared for lineage classification into Beijing (40) and non-Beijing(12) genotypes, and for epidemiological correlations. Results: Both MIRU-15 and 24 resolved to identify clearly distributed clusters. MIRU-12 presented incorrect relationship between Beijing and non-Beijing genotypes, and mixed up different lineages in the same cluster. MIRU-24 misinterpreted 1 non-Beijing cluster within 2 Beijing clusters. Epidemiological linkage within MIRU-15 and MIRU-24 clusters confirmed similar linkages between isolates. With MIRU-12, however, a few isolates showed incorrect linkages. Overall, MIRU-15 displayed more accurate differentiation when compared with MIRU-12 and 24. Conclusion: Our results showed that excessive or insufficient number of loci may reduce resolution of epidemiological links and clustering. In our experience, the 15-loci set for MIRU-VNTR outperforms the other two sets if used for routine genotyping. Although MIRU-15 is sufficient for simple clustering analysis, lineage classification and epidemiological linkages would need to be considered. Multiple methods and increase in number of highly variable loci may be indicated in order to achieve optimal specificity. Clostridium difficile is a gram-positive, spore forming anaerobic human pathogen. C. difficile infection (CDI) is a worldwide health concern with disease symptoms ranging from diarrhea, pseudomembranous colitis, and in some cases toxic megacolon, sepsis or death. CDI usually occur as a result of intestinal dysbiosis and exposure to C. difficile spores. Current treatment strategies rely on antibiotic usage which can often result in high recurrence rates and might increase the risk of developing antibiotic resistant strains. Previous researches have demonstrated that many medium chain fatty acids (MCFAs) can inhibit the growth of multiple human bacterial pathogens. Consequently, we hypothesized that these MCFAs might also present a potential inhibitory effect against C. difficile. Here we showed that lauric acid have potent antimicrobial activities against multiple toxigenic C. difficile isolates while presenting low cytotoxicity to human colon epithelial cells. The mechanisms of inhibition are due partly to the generation of the reactive oxygen species (ROS) and cell membrane damage. Biofilm formation and pre-formed biofilm were also significantly reduced by the addition of lauric acid in a dose-dependent manner. In murine infection model, lauric acid pre-treatment lessened CDI symptoms and reduced proinflammatory cytokine production. Taken together, we have demonstrated the naturally occurring MCFA lauric acid as a novel C. difficile inhibitor which could be used as an alternative treatment for CDI. Fluoroquinolones (FQs) are potent and broad-spectrum antibiotics commonly used to treat severe or resistant bacterial infections. However, FQ resistance to gram-positive and -negative bacteria has emerged easily and increased globally. The mechanism for FQ resistance is known to be multifactorial, including one or more mutations in their target genes encoding DNA gyrase and topoisomerase IV. Since the therapeutic choice against FQ-resistant bacterial infections becomes limited, it would be necessary to develop novel antibiotic alternatives to minimize or inhibit FQ resistant bacteria. In this study, we examined possible bactericidal effect of antisense peptide-peptide nucleic acids (P-PNAs) which were able to block the expression of DNA gyrase and/or topoisomerase IV in FQ-resistant Escherichia coli (FRE). The results demonstrated that a set of antisense P-PNAs, ASP-gyrA1 and ASP-parC1, targeting the translational initiation sites of DNA gyrase or topoisomerase IV genes showed the strongest inhibitory effects on the growth of FRE isolates. In addition, ASP-gyrA3 and ASP-parC2, which bind to the FRE-specific coding sequence within the gyrA and parC genes, respectively, possessed the selective bactericidal effects against FRE isolates. These results imply potential of those antisense P-PNAs as antibiotic alternatives against FQ-resistance bacteria. The resistance to fluoroquinolones is a main subject for the use of antibiotic substances in veterinary medicine where the overuse and misuse of antibiotics is a major cause of resistance. Optimizing the dose and dose intervals are essential for achieving clinical cures and minimizing the emergence of fluoroquinolones resistance. The purpose of this study was to evaluate pharmacokinetic/pharmacodynamic (PK/PD) parameters for the establishment of a reliable dose of enrofloxacin in chicken against Salmonella enteritidis. Male and female broiler chickens of 3-4 weeks old were assigned for this study. Enrofloxacin with a dose of 10 mg/kg of body weight were administered through per oral (PO) and intra-venous (IV) routes. Blood samples were collected at different time intervals and serum concentrations of the drug were determined. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of enrofloxacin in different strains of S. enteritidis were determined. The pharmacokinetic data were then integrated with pharmacodynamics data to determine the surrogate markers of antibacterial activity. The elimination half-life (T 1/2 ) of enrofloxacinwere 12.84 ± 1.4 and 25.84 ± 1.4 hr after IV and PO administrations respectively. The peak concentration (C max ), the time when the C max reached (T max ) and the area under the concentration-time curve (AUC) were 6.74 ± 0.03 µg/kg, 0.22 ± 0.1 hr, 21.13 ± 0.9 µg · hr/ mL for IV and 3.82 ± 0.59 µg/kg, 0.65 ± 0.12 hr, 20.84 ± 5.0 µg · hr/ mL, respectively. The bioavailability (F) of enrofloxacin after PO administration was 98.6 ± 8.9%. The MIC 50 of this drug against different strains of S. enteritidis were found to 0.50 µg/mL. When this pharmacodynamic data was integrated with pharmacokinetic data, the 24 hr area under the concentration-time curve (AUC 0-24 )/ MIC 50 were 42.26 ± 0.3 and 41.68 ± 0.1, and the C max /MIC 50 were 13.48 ± 0.7 and 7.64 ± 0.2 respectively for IV and PO administrations. It is concluded that enrofloxacin dosage of 50 mg/kg of body weight by IV and PO administration with 24 hr dosing interval will provide effective treatment for the infection of chicken by S. enteritidis. This data indicate that dosage of enrofloxacin upswing 5 times than the conventional recommended dosage. Introduction: Respiratory and gastrointestinal diseases are important causes of morbidity and mortality in livestock, leads to economic losses. Continuous monitoring of antimicrobial susceptibility of veterinary pathogens is needed because abuse and misuse of antimicrobials cause the emergence of antimicrobial resistance. In this study, isolates from cattle and pigs were tested for their antimicrobial susceptibilities. Materials and Methods: From 2011 to 2016, 1,459 respiratory and gastrointestinal isolates were collected from cattle and pigs, nationwide. In cattle, Trueperella pyogenes (n = 7), Pasteurella multocida (n = 11), Mannheimia haemolytica (n = 9), Clostridium difficile (n = 35), C. perfringnes (n = 116), and Escherichia coli (n = 176) were collected. Isolates originated from pigs as follows; T. pyogenes (n = 50), P. multocida (n = 149), Actinobacillus pleuropneumoniae (n = 61), Haemophilus parasuis (n = 15), Streptococcus suis (n = 177), Salmonella Typhimurium (n = 63), C. difficile (n = 170), C. perfringens (n = 214), and E. coli (n = 206). Antimicrobial susceptibility testing was performed using the disc diffusion method. Results: Overall antimicrobial resistance patterns were similar between cattle and pigs isolates. Most of the isolates showed relatively high resistance to majority of antimicrobials. E. coli and Salmonella exhibited low resistance to ceftiofur (3.5-12.6% ) and colistin (0-3.6%). Clostridium sp. was susceptible to amoxicillin (0-5.1%), florfenicol (4.1-9.4%) and doxycycline (1.2-10.8% ). Respiratory pathogens have relatively low resistance to ceftiofur (0-6.7%), cephalexin (1.3-13%) , amoxicillin (0-5.4%) and doxycyclin (0-18%). Conclusions: This study was conducted to investigate the antimicrobial resistance of bacterial pathogens in South Korea. These result may be provide the recent knowledge to establish strategies for treatment against respiratory and gastrointestinal disease in cattle and pigs. Abstract: The use of lactic acid bacteria as probiotics in food is on the increase in view of their ability to positively influence health and healthy living. In this report, we mined some of the bioinformatics of the strain, Streptococcus thermophilus KLDS 3.1003. 16s RNA sequencing was also carried out to determine its closest neighbor strains. S. thermophilus KLDS 3.1003 has a 1,899,956-bp genome with an exopolysaccharide production is 358 mg/L. Its closest neighbours include S. thermophilus LMD-9, LMG18311 and CNRZ1066. In depth amino acid coding gene analysis shows that S. thermophilus KLDS 3.1003 can synthesize all twenty (20) essential amino acids required for proper body functioning as well as animal feed formulations. These details are important in not only understanding the molecular basis behind its potential probiotic properties but also give new insights as to its potential future applications in the dairy and allied industries. Background: The components of our diet have been shown to have strong correlation with the spread of a number of diseases. In recent years and especially with the rapid development of 'omic' molecular tools, a large number of beneficial bacterial strains have been sequenced, thus increasing our understanding of the basis for their current and potential health benefits. As the second most important lactic acid bacteria in the world, several strains of Streptococcus thermophilus have been shown to have a number of in vitro and in vivo probiotic properties. We have recently shown that S. thermophilus KLDS 3.1003 possesses a total of 23 EPS-coding genes but also has high antimicrobial activity against pathogenic S. aureus, E. coli and G. vaginalis. Here, we report more data on the bioinformatics of this strain which may be linked to its potential probiotic properties. Methodology: S. thermophilus KLDS 3.1003 strain was obtained from traditional yoghurt starter culture in Inner Mongolia, China. The whole genome sequencing of Streptococcus thermophilus KLDS 3.1003 was performed using Pacbio RSII (20 K library) and Illumina Hiseq 4000 (500bp PCR-free library) strategies respectively. Then, 402 M Hiseq and 556 M Pacbio clean data were generated using a refined data filter. PacBio reads were assembled using the protocol in SMRT Analysis v2.3.0 Pipe: RS_HGAP_Assembly3. In addition, a phylogenetic tree was constructed to show the evolutionary status of S. thermophilus KLDS 3.1003 based on 16s rRNA analysis. EPS quantification of this strain was determined as earlier. Results and Discussion: In this report, we show that S. thermophilus KLDS 3.1003 displayed a similarity of ≥99% to Streptococcus stains deposited in NCBI based on 16s rRNA analysis. A distinctive branch of KLDS 3.1003 with Streptococcus thermophilus strains in the phylogenetic tree (see Figure 1 ) and S. thermophilus ATCC 19258 was found to be the closest evolutionary relative of KLDS 3.1003, thus resulting in the strain being recognized as S. thermophilus ssp. Other close neighbours include via MG-RAST analysis include S. thermophilus LMG18311 and S. thermophilus CNRZ1066. These strains have been studied extensively in a previous investigation. The genome of S. thermophilus KLDS 3.1003 consists of a 1,899,956bp chromosome with a G + C content of 38.90% (see Figure 2 ). Here, we have also analyzed its tRNA status and our result shows that S. thermophilus can synthesize all twenty (20) essential amino acids needed for proper body functioning in both adults and children. The EPS production of this stains was shown to be approximately 358 mg/L. EPS has been shown to have probiotic properties in recent reports. Further studies into the characterization of its EPS as well as its optimization are required to further explore the probiotic potentials of this strain. The amino acid profile of this strain confirms that it has the capacity to secrete all twenty (20) essential amino acids which are necessary for proper body functions in both children and adults. More amino acid-coding genes were found to be associated with the production of leucine, lysine, methionine, alanine and valine. The presence of lysine and methionine are key in animal feed formulation. This strain thus has potentials in the food industry as a functional food ingredient and starter culture. Further studies on the conditions for the optimization of its amino acid production are recommended. The presence of the Cl. difficile toxin gene (tcdA, tcdB and binary toxin) was also detected by PCR. Results: From the 81 fecal samples, ETEC were isolated from two suckling and eleven weaning pigs. The prevalence of LT/STb, STa/STb and STb in the thirteen E. coli isolates from diarrheic samples was 46.2% (n = 6), 30.8% (n = 4) and 23.1% (n = 2), respectively. Only one E. coli isolate from non-diarrheic sample was observed LT and STb toxin. Cl. difficile isolates (n = 9, 31.0%) from diarrheic samples were observed tcdA, tcdB and binary toxin. Conclusions: This study revealed that LT/STb toxin genes were commonly present in ETEC isolated from weaning pigs with diarrhea. And then C. difficile isolates from diarrheic fecal samples of suckling pigs were observed tcdA, tcdB, and binary toxin. Therefore, these results suggest that Cl. difficile and ETEC producing tcdA, tcdB, binary toxin and LT/STb toxins are important factor of diarrhea in piglets in Korea. Background: This study aimed to compare the PCR methods with serum agglutination test (SAT) in the diagnosis of brucellosis in patients before and six months after treatment. Methods: Peripheral blood specimens from 50 patients with brucellosis (case group), and 30 subjects without brucellosis (control group) were selected and entered into the study. The diagnosis of brucellosis was established using SAT ≥1:160 and 2mercaptoethnol (2-ME) ≥1:80 with clinical signs and symptoms compatible with brucellosis. Before treatment, SAT, 2-ME and PCR were done for each case and 6 months after completion of therapy. Subjects in the control group were assessed by the same tests at the initial visit. Results: In the case group, fifty patients (36 males, 14 females) with the mean age 43.6 ± 14.5 years were evaluated. The mean age of the control group was 40.6 ± 14 years. Among the 50 patients whose nested PCR assays were initially positive, 43 (86%) were negative six months after treatment. Relapse occurred in 5 (10%) patients within six months after treatment and all were PCR positive. None of the patients in the control group was PCR positive. The results show that PCR seems to be highly sensitive and specific and therefore a useful method for both the initial diagnosis and detection of relapse or chronic brucellosis. Background: The plasmid-borne mobile colistin resistance gene mcr-1 was initially identified in China, and was subsequently reported worldwide. Recently, mcr-1-harboring plasmids were identified in 11 Escherichia coli isolates from livestock in Korea. We investigated the characteristics of colistin-resistant E. coli with mcr-1-harboring plasmid Methods: Determination of growth rate and competition assay were performed to compare the bacterial fitness between mcr-1 plasmid harboring colistin-resistant E. coli (EC006, EC019, and EC111) and non-plasmid mediated colistin-resistant E. coli (E015R, E139R, and E154R) strains. Biofilm-forming capacity was assessed by microtiter plate method. For non-plasmid mediated colistinresistant strains, mutations and expression levels of the basRS, phoPQ, and eptA genes were determined. Lipid A structure was analyzed by matrix-assisted laser desorption/ionization time-offlight (MALDI-TOF) mass spectrometry. Results: Competition with E. coli MG1655 strain revealed no difference in competitive fitness between plasmid mediated and non-plasmid mediated colistin-resistant E. coli strains. Biofilm assay also showed similar levels of biofilm formation in all colistinsusceptible and -resistant strains tested. Three non-plasmid mediated colistin-resistant strains had amino acid alterations in BasS and their expression levels of eptA were significantly increased compared to those of colistin-susceptible strains. Lipid A structure in all mcr-1 plasmid harboring colistin-resistant strains was modified through the addition of phosphoethanolamine as in a non-plasmid mediated colistin-resistant strain E015R. Conclusion: Our data indicate that plasmid-borne mcr-1 does not impair fitness in E. coli. Lipid A modification by Mcr-1 was also the main colistin resistance mechanism, along with EptA and the spread of this transferable colistin-resistant determinant may accelerate the increase of colistin resistance. Background: Persister cell, which stays in dormant form when antibiotics exist, makes it difficult to remove pathogens completely from patients, which results in treatment failure. We characterized persister formation in Acinetobacter baumannii, one of the major nosocomial pathogens. Materials and Methods: In vitro antimicrobial susceptibility testing was performed with 20 clinical isolates of A. baumannii. Persister assay was performed with four antibiotics; colistin, amikacin, imipenem, and ciprofloxacin. To characterize properties of persister formation fromA. baumannii, persister assay was performed with various ways. Serial persister assay was also performed; two random persister colonies which were survived from first persister assay, were re-inoculated into fresh LB broth each and incubated overnight. Second persister assay was performed for the overnight cultures, with the same protocol. Results: For four antibiotics, there was no correlation between persister rates and specific strains. That is, an isolate exhibiting a high persister cell formation rate for one antibiotic did not necessarily show a high rate of persister production for other antibiotics. The persister formation rate for the same antibiotic was almost constant when the persister assay was repeated three times. Serial persister assay for re-grown persister cell in antibiotic-free media showed that the persister formation rate was the same as the initial one. Conclusions: Our data suggest that persister cells are formed by different mechanisms for each antibiotic. The persister rate against antibiotics is an intrinsic property of each strain. Carbapenem has been used for treatment against bacterial infections as the most reliable last-line drugs. However, with the abuse of carbapenem, carbapenemase-producing Enterobacteriaceae (CPE) as well as Carbapenem-resistant Enterobacteriaceae (CRE) has emerged in worldwide. In Korea, all laboratories should submit suspected CRE isolates to KCDC for confirmation of CRE and carbapenemase-production. In this study, we confirmed and analyzed characteristics of CRE isolates like species identification, carbapenem resistance patterns and carbapenemase gene type. Antimicrobial susceptibility was examined by broth microdilution and disk diffusion methods using CLSI guideline. The carbapenemase genes (IMP, OXA-48, VIM, NDM, KPC and GES) were investigated by PCR and analyzed by sequence alignment method. A total of 1,810 CRE suspected isolates were collected (between January and June 2017) and 1,793 (99%) isolates were Enterobacteriaceae and 17 (1%) were non-Enterobacteriaceae. The ratios of CRE, CIE and CSE were 87%, 7% and 6%, respectively. Among CRE isolates (1,562 isolates), Klebsiella pneumonia, Escherichia coli and Enterobacter spp. were the most prevalent types of CRE isolates, with the occurrence in 65%, 14% and 14%. In addition, 986 isolates showed CPE positive (63%) and harbored bla KPC−2 and bla NDM−1 gene 703 (73.1%) and 142 (14.4%), respectively. Most of CPE had one carbapenemase gene but 38 (3.85%) isolates harbored 2 genes at the same time and predominant type was bla NDM−1 and bla OXA−181 type (21 isolates, 55%). In conclusion, June 3, 2017 mandatory submission system of suspected CRE was started in Korea, so continuous monitoring of CRE occurrence and characteristics change will be investigated and reported. Background: Fluoroquinolone is one of the most commonly used antibiotics for the treatment of infections caused by Streptococcus pneumoniae. However, the rates of fluoroquinolone resistance are increasing according to the frequent use. We designed this study to verify the current fluoroquinolone resistance rates and risk factors for community-onset pneumococcal pneumonia. Methods: A retrospective case-control study was conducted in a tertiary referral hospital. Study population was composed of patients admitted for pneumococcal pneumonia between January 2011 and May 2017. The case group included community-onset pneumonia caused by levofloxacin-nonsusceptible S. pneumoniae. The control group consisted of the 2 patients who were admitted around the same time as each case among the patients with levofloxacin-susceptible S. pneumoniae. Results: A total of 198 pneumococcal pneumonia were identified during the study period. 25 levofloxacin-resistant S. pneumoniae and 3 levofloxacin-intermediated S. pneumoniae were included as a case group. The nonsusceptible rate to levofloxacin was 14.1%. Multivariate analysis showed that recent hospitalization (OR 5.40, 95% CI 1.35-21.58, P = 0.017), underlying bronchopulmonary disease (OR 6.03, 95% CI 1.47-24.81, P = 0.013), cerebrovascular disease (OR 8.44, 95% CI 1.37-52.11, P = 0.022), and prior antibiotics use within 3 months (OR 6.90, 95% CI 1.89-25.22, P = 0.003) were associated with levofloxacin non-susceptibility. Conclusion: The independent risk factors for levofloxacin nonsusceptible pneumococcal pneumonia were recent hospitalization, bronchopulmonary disease, cerebrovascular disease, and prior antibiotics use within 3 months. More careful choices of empirical antibiotics are needed among these groups. The correlations between fraction inhibitory concentration index and mutant selection index in an antimicrobial combination X. Xu, L. Xu, G. Yuan*, J. Li. College of Bioscience and Bioengineering, Jiangxi Agricultural University, Nanchang, China Abstract: Antimicrobial resistance seriously threatened human health, and combination therapy was generally considered to be an effective strategy to prevent resistance. As its general benefits have been difficult to conclusively demonstrate, the validity of synergistic combination as we mentioned was probably a key. According to the hypotheses of mutant prevention concentration (MPC) and mutant selection window (MSW), combination in which mutant selection index (SI, the ratio of MPC in combination to MIC 99 alone) of each agent was less than or equal to one would be efficacious to prevent antimicrobial resistance. Are there some correlations between SI and fractional inhibitory concentration index (FICI) used to evaluate the synergy effects of antimicrobial agents? Using checkerboard assay and agar plates with linear concentration decrease, our researches on mutant SIs of five antimicrobial agents alone and their FICIs and SIs in three combinations including twenty-one different proportions against methicillin-resistant Staphylococcus aureus (MRSA) (Figure 1 ) indicated that: (1) The smaller the FICIs of two agents in a combination were, the more probable both SIs were less than or equal to one, and the wider the proportional range of two agents closing each other's MSW was; (2) Only the minimum FICI was less than or equal to 0.50, some combinations of two agents closing each other's MSW could be sought out from their different proportions; (3) Different proportions of two agents in a combination presented different SIs for each one, and their synergistic validity in vivo would be accordingly dicounted due to their different pharmacokinetics. So, synergistic antimicrobial agents with simliar pharmacokinetics for combinations must be favorable to prevent resistance, and their maximum FICIs were perfectly less than 0.50. periods, with the remarkable increase of ST166, which mainly due to increase of serotype 11A in Korea. In addition, ST558 which consisted of serotype 35B was also found frequently in 2012-2016, which was not found in [2008] [2009] . Conclusion: This study showed the persistently high prevalence of macrolide resistance and increasing number of pneumococci carrying both erm(B) and mef(A) in Korea. Given the current epidemiology of macrolide resistance, an empirical use of macrolides alone may not be an appropriate choice for the treatment of pneumococcal infection in adult patients in Korea. Therefore, continued surveillance of pneumococcal epidemiology of macrolide resistance would be necessary. According to the clinical and laboratory standards institute (CLSI) guideline, agar and broth dilution method were used to detect vancomycin resistant staphylococcus aureus. However, recent advances in automated devices have led to increased use of automated instrument in hospitals. In this study we compared the results of vancomycin susceptibility test between agar, broth dilution method and vitek2 instrument using vancomycin MIC 3∼4 ug/ml Staphylococcus aureus strains by E-test method. A total of 27 strains were used in this experiment which were collected in 2016 year through vancomycin resistant Staphylococcus aureus surveillance system in Korea. The accordance between agar dilution and broth dilution was 85% (4 strains were different from 27 strains). But resistant susceptibility was not affected. And the accordance between agar, broth dilution method and vitek2 instrument was 74%, respectively. The results of vitek2 instrument showed 2 times higher MIC (4 strains) and 4 times higher MIC results (1 strain) than CLSI method. These results changed vancomycin susceptible (MIC 2) to intermediate (MIC 4, MIC 8) susceptibility. Until now, the detail accordance analysis was not accomplished against vitek2 vancomycin susceptibility using large number strains of various vancomycin MIC susceptibility. But this study showed vitek2 results need additional broth dilution experiment for vancomycin resistant susceptibility confirmation. The BTB prevalence at animal and herd level were; 0.62% and 0.77% in beef cattle, 0.28% and 0.91% in dairy cattle, respectively. Overall, prevalence was 0.47% and 0.78% at animal and herd level, respectively. Five risk factors were offered to invariable logistic regression model. Farms with higher numbers of cattle, and those with one more positive animal, were 2.6 and 1.9 times more at risk for establishment of BTB, respectively, compared with the farm only one BTB outbreak. Conclusion: To our understanding this is the first report that has encompassed dairy and beef farms in South Korea. Furthermore, results of risk factor analysis will be useful as entry points for future informed decision making towards BTB control and eradication program in the country. Conclusion: Our data showed the LTBI prevalence among HCWs working in our hospital were 40%, and was increasing tendency by age. Physicians were less likely to accept treatment than other HCWs, and this finding suggests effort to enhance LTBI treatment acceptance among physicians will be needed. Background: Bovine tuberculosis (bTB) remains an important disease in many countries of the world, causing significant economic losses and proving difficult to control. The detection method of residual Mycobacterium bovis (M. bovis) in the environment is needed to control bTB on herd level. Nested PCR amplification was applied in this study to ensure the specificity of detection, eliminate any false-positive results, and increase the amplification signal, providing the method with the highest sensitivity of environmental sample. In this study, we attempted to establish a method for detection of M. bovis in environmental sample using nested-PCR method and to detect residual M. bovis in the cattle herds where was carried disinfection. Materials and Methods: 60 Environmental samples (51 fecal samples, 9 bottles water) from (10 herds) herds where have a history of bTB outbreak were used for the detection of residual Mycobacterium bovis. Amplicon libraries of IS 6110 target gene fragment was amplified from M. bovis DNA by using the two types of primer sets. For the first step, PCR amplification of the 716 bp length for IS 6110 gene was performed and each sample was purified to remove the others. The purified library amplicons were re-amplified by second PCR primer set (203 bp) to increase detection sensitivity. After each PCR amplification round, the size of the PCR product was verified on a 1.5% agarose gel. Result: The results of detection of residual M. bovis on environmental sample, 48 fecal samples DNA were shown positive band by nested PCR method and 8 samples of water samples were detected as positive. The results of the detection of M. bovis using Nested PCR method were found to be higher than that of the commonly used PCR method. The sensitivity and specificity of two methods based on the polymerase chain reaction (PCR) for detection of M. bovis were compared. In the standard PCR method, the sensitivity of detection of M. bovis was very low due to contaminants and other microorganisms in environmental samples. However, the sensitivity of this nested-PCR for M. bovis was high compared to previous one, which is useful as a detection method in environmental samples. Due to resistance to outside environment, bTB is often reported as recurrent outbreak in same herd. Therefore, it is important to carry out more thorough hygiene management with test and slaughter of reactors. This nested-PCR will be used as a useful tool for screening Mycobacterium bovis in environmental samples as one of the methods to control bTB on herd level. (26 cases) were the most common sites of TB presentation. CNS involvement was the third most common presentation (25 cases), followed by TB lymphadenopathy (21 cases), pleurisy (21 cases), pericardial involvement (11 cases), skin involvement (2 patients), and others (5 cases). Of the 163 patients, 114 (69.9%) had positive QuantiFERON-TB Gold assay result, and 2 (1.2%) had indeterminate result. The percentage of positive QuantiFERON-TB Gold assay results was highest in the proven TB group (41/47, 87.2%), followed as 69.1% (38/55) in probable TB group, and then 57.4% (35/61) in possible TB group (p = 0.007). In the 47 proven cases, 5 cases (10.6%) had negative QuantiFERON-TB incubation at 37°C for an hour, anti-mouse-antibody (1:5000, SIGMA A 6154) was added. Lastly, TMB (BD Biosciences) was added to each well (100 μl/well). Then, 50 μl of the Stop solution (2N H2SO4) was added to each well. Finally, the optical density was measured at 450 nm. ELISA: The purpose of this ELISA was to observe the binding of mRID fused RBD to the MERS-COV infected rabbit and human serum. MERS-infected cell lysate (MERS-CoV/KOR/KNIH/ 002_05_2015), mRID fused RBD and HR2 were loaded onto 96well nunc immunoplate (Thermo Fisher Scientific) for O/N coating at 4°C. At the end of each procedure, there were washed with 200 ul of PBST three times. The blocking buffer (1% BSA in PBST) was treated with 200 μl/well. After allowing two hours of reaction time at RT, MERS-COV infected rabbit sera (starting from 10 ug to 0.25 ug) and the Patient serum (1:100 diluted) were placed into the wells (100 ul/well). After two hours of incubation at RT, diluted 2nd antibody (1:3,000, Goat Anti-human IgG Antibody, KPL) was added, measuring 100 ul per well. After another hour of incubation at RT, TMB (BD Biosciences) was added at 50 μl/well were added. Further reaction was stopped by adding 50 μl of STOP solution (2N H 2 SO₄) into the well. Finally, the optical density was measured at 450nm. Target protein design, cloning, expression of target protein using RID as a fusion partner: To express MERS-CoV antigen in E. coli, 70 amino acid RID amongst the LysRS was used as the fusion partner ( Fig. 2A) . Depending on the species of immunization, hRID and mRID were used for cloning to minimize any induced immunity for the fusion partner. The following scheme is a simple illustration of a RNA-mediated folding model that produces only the folded target proteins by cleaving fusion partners using the TEV protease (Fig. 1A) . HR2 was selected based on the antigenicity plot prediction. From the previous study, the RBD that plays a key role in the binding of dpp4 receptor was selected as the target protein (Fig. 2B) . mRID (mouse-derived RID) was used as a fusion partner for expression. Its solubility improved as the temperature decreased when compared to the solubility of the direct form (Fig. 3C) . Folding of the RID-fused protein and the role of RNA in stability maintenance: According to former studies, many people believed that most protein folding was carried out by molecular chaperones. However, recent research showed that RNA can function as chaperones to aid in protein folding and stability. Thus, we carried out cell lysis to verify the role of RNA in RID-fused proteins in the folding and stability maintenance. RNA was removed in-vitro, while another was incubated as the control at 37°C. Agarose gel verified that RNA was removed only when treated with RNase A (Fig. 3A ). In addition, the SDS-PAGE analysis using Western blot and Coomassie staining confirmed that both mRID fused protein showed a decrease in their solubility when RNA was removed (Fig. 3B) . RID fused RBD protein bound to hDPP4 receptor: We verified the binding between the RBD protein produced in the E. coli system and the hDPP4 receptor using the co-immunoprecipitation and ELISA. First, the binding between RBD protein and hDPP4 was verified using co-immunoprecipitation by protein A magnetic beads with dpp4-specific antibody. Then, we performed Western blotting (Fig. 4A) . Also, we used hdpp4 protein as the coating antigen to observe that binding of the RBD protein is concentration-dependent. Then, antihis antibody was used in the ELISA assay for further verification (Fig. 4B) . Such findings concluded that successful folding of E. coli derived RBD protein result in binding to the DPP4 receptor. Sero-diagnosis based on the RBD and HR2 antigen from MERS-COV infected serum: We predicted that the mRID fused RBD and HR2 protein are suitable for sero-diagnosis. Thus, RBD was used as the coating antigen, the rabbit sera immunized with inactivated MERS-COV was serially diluted first for the ELISA assay. We identified binding of the RBD and MERS-COV immunized rabbit sera (Fig. 5A ). Furthermore, ELISA was performed using the sera (1:100 diluted) of the four patients infected with MERS-COV. For the RBD coating antigen, the binding sensitivity for the three MERS-COV infected patient sera is higher than the MERS-COV infected cell lysate (Fig. 5B ). However, for the 1:100 diluted normal person sera, almost none was detected by the RID-fused RBD and HR2. These results demonstrate that mRID-fused RBD and HR2 can serve as a powerful antigen for rapid and accurate diagnosis and mass production. Preceding research papers about MERS-CoV introduced the method of producing spike protein from insect cell using target epitopes. However, the drawbacks of such preexisting method include high cost and low production output. Therefore, it is certainly necessary to mass-produce the appropriate diagnostic antigens at a low cost and time efficient manner. In conclusion, for the first time, we employed Chaperna (RNAmediated chaperone) to produce potential sero-diagnostic antigens such as RBD and HR2 from E. coli to diagnose patients and animals infected with MERS-CoV. Using ELISA, it was verified that the antigens bind to the patient's serum with a higher affinity than to the MERS-CoV infected cell lysate. Also, we were able to establish a high accurate diagnostic technique using an additional HR2 antigen to double-check the verified diagnosis. This platform will become extensively applicable not only to MERS-CoV, but also to the outbreak of various pandemic viruses to allow quick diagnosis. We continue our endeavor for improving and applying the RNAdependent folding system for diagnostic and prophylactic applications. Ongoing research is dedicated to harnessing Chaperna for the assembly of vaccine antigens into nanoparticles for gaining high neutralizing capacities. Severe fever with thrombocytopenia syndrome associated with hemophagocytic lymphohistiocytosis: review of 7 cases with bone marrow study S. Park 1 , J. are few studies about electrocardiographic (ECG) changes and myocardial dysfunction. Methods: We studied 34 patients who were diagnosed SFTS in five hospitals in Daegu, Korea from January 2013 to November 2016. We reviewed the medical record retrospectively. Result: Abnormal ECG changes were observed in 20 patients (58%) during hospitalization. Among the 20 patients, arrhythmic changes were 32% (11/34), ischemic changes were 26% (9/34): atrial fibrilliation 6, atrial flutter 4, paroxysmal supraventricular tachycardia 1, T wave inversion 9. Normal sinus rhythm also changed during hospitalization in 8 patients who were taken next ECG (34.7%, 8/23). The patients who had abnormal ECG were older than normal ECG group, also had pulmonary edema more frequently. ECG abnormalities occurred more frequently in the death group. (72.7% vs. 52.5%, p = 0.295). Conclusions: This study showed that high proportion of SFTS patients had ECG changes (58%). The cardiac involvement of SFTSV was not clear. But, lots of patients were having abnormal ECG and more frequently involvement in fatal cases. Additional studies are needed about cardiac involvement of SFTSV. Since the outbreaks of Hendra virus and Nipah virus in Asia, several Henipaviruses detection assays based on real-time reverse transcription polymerase chain reaction (rRT-PCR) have been developed. In this study, analytical validation of these assays was performed to evaluate their sensitivity and reliability for the preparedness of Henipavirus outbreak. The assays targeting Hendra virus matrix gene (M) and Nipah virus nucleoprotein (N) gene were evaluated for sensitivity by calculation of limits of detection (LODs), estimated among over 13 replicates with 0.01-100 copies/reaction. The assays targeting both Hendra virus and Nipah virus phosphaprotein gene (P) were also evaluated. The performances of these assay systems were comparable to each other when tested against in vitro transcribed RNA samples. Our results demonstrate that these assays are reliable tools for the rapid diagnosis, clinical surveillance, and epidemiological study of suspected Henipavirus cases. Formerly these samples were tested for influenza viruses by RT-PCR. Full length of hemagglutinin gene of the isolates was sequenced using Sanger method. Also 118 samples including previous 29 isolates, full genome segments were obtained on a MiSeq platform (Illumina). Phylogenetic tree was generated using MEGA v.6 and correlation plots were constructed between each amino acid substitution. Phylogenetic tree analysis revealed that the isolated Ainfluenza viruses belonged to the subclades of 3C.2a. A correlation plot using 34 amino acid substitutions in HA from our isolates revealed that the R142K and R142G substitutions were positively and negatively correlated with the D160N substitution, respectively, suggesting that the R142K and R142G were obvious characteristics of viruses in the new subgroup. Also full genome segments from MiSeq platform were revealed various correlation result. Isolated Viruses period in 2016-2017 season, more than 30% of sample had 142 substitution mutation, which is one of the key characteristics of 3C.3a clade represented by the A/Switzerland/9715293/2013 virus, the vaccine strain in the previous season. Mortality was assessed at 7 days from the start of antibiotic use. Patients discharged or transferred to other medical institutions within Mortality was assessed at 28 days from the start of antibiotic use. Patients discharged or transferred to other medical institutions within 28 days were excluded from the analysis because they could not be assessed for survival. Patients who were discharged within 28 days, but after the antibiotic therapy for CRAB was terminated and the condition improved Depending on the combination of antibiotics used, they may belong to more than one classification Methods: Experiments were performed with two S. epidermidis strains, RP62A and IDRL-8883. RFP-resistant mutants were selected in vitro by serial passaging with progressively increasing 2-fold concentrations of RFP. Both IDRL-8883-s (susceptible to RFP) and 8883-r (resistant to RFP) were recovered from a patient with a prosthetic joint infection. RP62A 3Bs, 3Br, 1Ws, and 1Wr were recovered from bone (3Bs and 3Br) or a foreign body (1Ws and 1Wr) of animals identically infected with RP62A and treated with RFP monotherapy. A total of 16 colonies with a RP62A background and a total of 63 colonies with anIDRL-8883 background were analyzed for rpoB mutations. The relative fitness of the RFP susceptible and the isogenic RFP resistant strains were determined using a paired competition assay. Results: A resistant strain was rapidly selected in a single-step fashion with RP62A. In contrast, resistance to RFP was observed in incremental steps with IDRL-8883. All mutations were detected in cluster I of rpoB. The five following amino acid substitutions were detected in vitro: Asp-471→Asn, Asp-471→Gly, Asp-471→Val, Ser-486→Tyr, and His-481→Tyr. The three following amino acid substitutions were detected in vivo: His-481→Tyr, Ser-486→Phe, and Gln-468→Lys. in vitro competition assays revealed that all RFPresistant mutants other than Ser-486→Phe and Ser-486→Tyr had a relative fitness of less than 1.0. Prolonged exposure to RFP selected for RFP-resistant strains (Ser-486→Phe and Ser-486→Tyr)with no fitness defect. Conclusions: Several point mutations concentrated in cluster I of rpoB emerged over the course of exposure to RIF; some, but not all of these mutations were associated with fitness defects. P2-LB37 Antimicrobial resistance of Streptococcus pneumoniae isolates from adult patients with invasive pneumococcal disease or pneumonia The primary specimen sources were sputum (73.7% and 54 Macrolide resistance was persistently high (62.8% and 68.6% in 2008-2009 and in 2012-2016, respectively) with the highest in China (89.1% and 90.8, respectively) and Korea (74.2% and 79.7%, respectively). Multidrug resistance (MDR) was observed in 49.5% and 52.9% of isolates in Siriraj Hospital, Thailand, 5 Peking University People's Hospital, China, 6 Research Institute for Tropical Medicine, Philippines, 7 Institute for Medical Research, Malaysia, 8 Ghangi General Hospital Non-PCV serotypes has markedly increased from 35 54) for VRE. MDRO acquisition included increasing age and current use of antibiotics. One patient died of gastrointestinal bleeding and none developed infection from colonization at a median of 3.5 months of follow up (IQR: 2.4-4.5). Conclusions: The prevalence and incidence for MDRO colonization in the general ward is higher than expected. This may impact on future strategies to successfully prevent and control MDRO. P2-LB40 Presence and characterization of mcr-1 gene and carbapenemase gene in sewage water of five tertiary-care hospitals in Beijing Longyang Jin 1 † , Ruobing Wang 1 † , Xiaojuan Wang 1 , Qi Wang 1 , Yawei Zhang 1 Results: Nine MCRPE and 12 CPE isolates were obtained. All mcr-1 −75 kb, and 30−90 kb, respectively. Conclusions: To the best of our knowledge, this is the first report of mcr-1-positive E. coli and bla NDM-1 -carrying E. cloacae and C The rank order of occurrence and antifungal period (from October Results: The Candida species isolated from all clinical specimens (340 isolates) included Candida albicans (161, 47.4%), Candida glabrata (58, 17.1%), Candida tropicalis (57, 16.8%), Candida parapsilosis (46, 13.5%), Candida krusei (6, 1.8%), Candida guilliermondii (3, 0.9%), Candida auris (2, 0.6%), Candida lusitaniae (2, 0.6%), and other 5 isolates. In comparison, the most frequently recovered Candida species from blood and other sterile fluids (241 isolates) was C. albicans (102, 42.3%), C. tropicalis (54, 22.4%), followed by C. glabrata (48, 19.9%), and C. parapsilosis (27, 11fluconazole (one C. albicans, one C. glabrata, one C. tropicalis, three C. parapsilosis, and six C. krusei isolates), 0.7% for voriconazole (one C. tropicalis and one C. krusei isolates), 0% for micafungin, and 0.3% for caspofungin (one C. glabrata isolate), respectively. All isolates were WT to amphotericin B. Among 12 isolates of non-common species, four (two C. auris, one C. guilliermondii, and one Candida pelliculosa isolates) showed a fluconazole MIC of ≥64 µg/ml. All these isolates with azole or caspofungin resistance were recovered from sterile sites except two C. auris isolates. No significant differences in MIC 50 and MIC 90 were detected between the blood Candida isolates and those collected at other sites for each of the five antifungal agents. Conclusions: The current study shows that overall resistance rate of Candida species is still low in Korea. Continuous national surveillance programs should be needed to identify changes in the antifungal susceptibility patterns of Candida isolates with caution to increasing C. glabrata having the potential for resistance to antifungal agents. Keywords: Candida, Distribution, Antifungal susceptibility, Multicenter study P2-LB44 Introduction of one health approach to antimicrobial Acknowledgements: This study was supported by the Acknowledgements: This study was supported by the Korea Acknowledgements: This work was supported by grants from the National Natural Science Foundation of China (No. 81460529, 81660578 and 81260476). Acknowledgements: THis study was supported by the Korea Centers for Disease . Keywords: Staphylococcus aureus, vancomycin resistance, VRSA, VISA tRNA-Leu-CAG tRNA-Leu-AAG tRNA-Leu-TAG tRNA-Leu-TAA tRNA-Leu-CAA tRNA-Leu-TAG tRNA-Leu-TAA tRNA-Leu-TAG This project is expected to be extended with the participation of related ministries with building an interministerial frame work for AMR, and the strategic investment in R&D such as development of diagnostic tools and control technologies for AMR will be increased. One Health approach to AMR may produce meaningful data to enable analysis of the occurrence and transmission of AMR, and establish prevention and control programs against AMR. Background: Bovine colostrum is the first milk produced by cows during the initial days after giving birth. Immunoglobulin G (IgG) is the most predominant immunoglobulin in colostrum of cows. This study investigated prevalence of IgG in bovine colostrum and colostrum products.Methods: A total of 15 colostrum samples were collected from healthy bovine (n = 5) at the first (within 3h), second (12h), third (24h) milking postpartum. A total of 12 colostrum replacement products were collected from domestic farms. The concentrations of IgG in the all samples were measured by commercial bovine IgG ELISA kits following the procedures recommended by the manufacture.Results: The IgG concentrations of the colostrum by milking time ranged from 32.9-68.5 mg/ml, 7.6-53.3 mg/ml and 3.8-25.7 mg/ ml, respectively. The IgG of colostrum replacement products reveled that dry colostrum (62.4 mg/ml), frozen colostrums (38.7, 35.8, 10.8 mg/ml), immune booster ( powder; 11. 2, 9.8, 9.2 Gold assay. Among the 47 patients with gastrointestinal involvement, QuantiFERON-TB Gold assay was positive in 39 (83%), 16 (61.5%) were positive in skeletal TB, and 100% in with disseminated extrapulmonary TB. The sensitivity in the proven and probable group was 79%, specificity was 42.6%, positive predictive value was 69.2% and negative predictive value was 44.6%. Conclusion: QuantiFERON-TB Gold assay was not useful diagnostic method for extrapulmonary tuberculosis in BCG vaccinated intermediate-TB endemic area. Despite of negative QuantiFERON-TB Gold assay result, physicians should not ignore the possibility of tuberculosis in patients with suspected TB clinically. The potential of an effective anti-tubercular candidate agent of next-generation, and Mtb inhibitory effect W.-H. Choi*. Department of Biomedical Science, and Medical Zoology, Kyung Hee University School of Medicine, Seoul, South KoreaBackground: Tuberculosis (TB) is a major infectious disease that causes the highest human mortality, particularly in Africa, around the world. This study was carried out to evaluate anti-Mtb effect of a compound (named WK2015001) that causes inhibitory activity against the growth and proliferation of Mycobacterium tuberculosis (Mtb).Methods: The anti-Mtb activity of WK2015001 was determined using different anti-Mtb indicator methods such as resazurin microtiter assay (REMA), Ogawa slant medium assay, and MGIT 960 system assay. The Mtb was incubated with various concentrations (0.5-16 µg/mL) of the compound and anti-Mtb first-line drugs for 6 days in the REMA, and for 4 weeks in MGIT 960 system assay.Results: WK2015001 indicated the anti-Mtb effect by strongly inhibiting the growth and proliferation of Mtb in a dose-dependent manner in the REMA, Ogawa slant medium assay and the MGIT 960 system assay. In particular, WK2015001consistently induced anti-Mtb activity by effectively inhibiting the growth and proliferation of Mtb in MGIT 960 system for 4 weeks with a single treatment, and its minimum inhibitory concentration was measured as 5 µg/mL. Conclusions: These results demonstrate that WK2015001 not only have the selective anti-Mtb effect/properties, but also induces effectively anti-Mtb activity by strongly inhibiting the growth and proliferation of Mtb through blocking signaling pathways of Mtb. Therefore, this study suggests the potential that the compound can be used as effective anti-Mtb candidate agents of next-generation for developing a new anti-tuberculosis drug against the global challenge of TB. When it comes to E. coli system, cell growth is fast, the medium price is affordable and expression levels are high. However, the absence of refolding by insolubility of produced protein makes folding impossible, a critical defection of such system. But we recently developed chaperna such as RID that enhance protein solubility and folding. We expressed RBD as target antigen instead of spike protein. Also, HR2 (heptad repeats) was expressed for target antigen and showed diagnostic effectiveness. These data demonstrate that using this RNA-dependent protein folding technology, diagnostic platform could be established through production of recombinant antigen of even a virus with a high casualty rate and pandemic occurrence. Expression vector construction, expression and purification of recombinant proteins: PCR was performed for the RBD and HR2protein using the S sequences (GenBank accession no. AFS88936.1) of MERS-COV. All plasmids which generate pGE-mRID were transformed into BL21pLysS Competent Cells. 50 ml of cultured cells were added to 500 ml of LB medium to induce protein expression at 16°C. RID-fused RBD and HR2 proteins were purified using the Histrap HP column (GE healthcare). The imidazole concentration gradient was increased for purification using the wash buffer and the elution buffer. Experiment to verify the folding and stability of RNA through in vitro elimination: To verify the RNA mediated folding and stability, 15 ml of cultured cells were harvested. Cell lysates were obtained using 500 ul of B-PER II (Thermo scientific, 78248). The cell lysate was centrifuged at 12,000 rpm for 10 minutes. The obtained soluble fraction was divided in half in which 250 μg/ml was treated with RNase A (iNtRON Biotechnology, 27062) and the negative control without RNase A was incubated at 37°C for 15 minutes. After the removal of RNA, a soluble fraction (SS) and a pellet fraction (SP) were collected through centrifugation at 12,000 rpm for 15 minutes. For the western blot, Penta His Antibody (1:5000, QIAGEN, 34660) was used and anti-mouse IgG (1:20000, Sigma, A4416). Hdpp4 binding assay: The binding between the MERS RBD protein and hadpp4 protein was observed. Each of the 96-well immunoplate (Thermo Fisher Scientific) with 5 ug/ml hdpp4 protein (abcam, ab79138) was coated at 4°C (100 μl/well). At the end of each procedure, they were washed in PBST three times. The blocking buffer (1% BSA in PBST) of 150 μl per well was stored at RT for an hour. After the ½ dilution of mRID fused RBD, 100 ul of each sample was added. After incubation at 37°C for 2 hours, anti-his antibody (1:1000, QIAGEN, 34660) was added. Then, after