BACKGROUND: At antimicrobial susceptibility testing (AST), clinical isolates may appear susceptible sometimes to the antibiotics that are clinically ineffective or due to technical errors in the testing. So an interpretive reading of AST should be done, but most hospitals do not perform it routinely. Here, we developed and evaluated a computerized expert system to interpret AST of Enterobacteriaceae to beta-lactam antibiotics. METHODS: We made a rule-based expert system according to the natural resistance of the members of Enterobacteriaceae and the common phenotypes of resistance mechanisms for Enterobacteriacae. Antimicrobial suceptibility testings were performed using the disk diffusion method with 12 beta-lactam antibiotics for a total of 1, 016 clinical isolates. Then we compared the raw and expert results of AST. RESULTS: An overall discrepancy rate due to natural resistance was 5.9%; 10.4% for Klebsiella spp. and Citrobacter diversus, 15.0% for Enterobacter spp., 2.6% for Serratia marcescens, 31.6% for Morganella morganii and Providencia stuartti. Accoriding to acquired antimicrobial resistant mechanisms, overall resistant discrepancy was 21.8%; 18.8% for Escherichia coli, Proteus mirabilis, Salmonella and Shigella spp., 25.9% for Citrobacter diversus and Klebsiella spp., 21.6% for Citrobacter freundii and Enterobacter spp., 45.6% for Morganella morganii, 10.0% for Proteus vulgaris, 12.2% for Serratia spp.. CONCLUSIONS: These results indicate that the application of the expert system for interpretation of antimicrobial susceptibility test may provide more reliable data for the treating physician. Additional information should be applied on the software for new resistant mechanisms or some misinterpretive readings.
Anti-Bacterial Agents ; beta-Lactams* ; Citrobacter freundii ; Citrobacter koseri ; Diffusion ; Enterobacter ; Enterobacteriaceae* ; Escherichia coli ; Expert Systems* ; Immunity, Innate ; Klebsiella ; Morganella morganii ; Phenotype ; Proteus mirabilis ; Proteus vulgaris ; Providencia ; Reading ; Salmonella ; Serratia ; Serratia marcescens ; Shigella

BACKGROUND: Cefroxadine is an oral first-generation cephalosporin, which has been used for several years. But, the susceptibility data of cefroxadine were rarely reported in Korea. The current study attempted to determine the antibacterial activity of cefroxadine against the major clinical isolates. METHODS: According to the NCCLS recommendations, antibacterial activities of cefroxadine were measured against total 500 major clinical isolates. MICs were determined by the agar dilution method, a series of doubling dilutions from 128 to 0.03 /mL, on Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Citrobacter freundii, Serratia marcescens, Proteus mirabilis, and Staphylococcus spp. In case of Haemophilus influenzae, Streptococcus pneumoniae, and Moraxella catarrhalis, broth microdilution method, a series of doubling dilutions from 16 to 0.015 /mL, was performed. RESULTS: Cefroxadine had variable activity against Enterobacteriaceae. MIC cumulative curves showed that cefroxadine had relatively low MIC distributions against E. coli, K. pneumoniae and P. mirabilis, showing MIC50 were 4, 4, and 8 /mL, respectively. Against E. cloacae, C. freundii, and S. marcescens, cefroxadine 's MIC50 values ranged from 128 to >128 /mL. For clinical isolates of methicillin-susceptible Staphylococcus aureus and methicillin-susceptible Staphylococcus epidermidis, cefroxadine had MIC90 values were 4 /mL and 8 /mL, respectively. Cefroxadine had MIC50 values of 1 /mL and >16 /mL for penicillin-susceptible and penicillin-not-susceptible strains of S. pneumoniae, respectively. Cefroxadine had MIC50 values of 8 /mL and 4 /mL against H. influenzae and M. catarrhalis, respectively. CONCLUSION: Cefroxadine had good activity against gram-positive bacteria, except penicillin-resistant S. pneumoniae, and showed moderate antimicrobial activity against M. catarrhalis, E. coli, P. mirabilis, and K. pneumonaie. Cefroxadine had variable activity against Enterobacteriaceae other than the above-mentioned species.
Agar ; Citrobacter freundii ; Cloaca ; Enterobacter cloacae ; Enterobacteriaceae ; Escherichia coli ; Gram-Positive Bacteria ; Haemophilus influenzae ; Influenza, Human ; Klebsiella pneumoniae ; Korea ; Mirabilis ; Moraxella (Branhamella) catarrhalis ; Pneumonia ; Proteus mirabilis ; Serratia marcescens ; Staphylococcus ; Staphylococcus aureus ; Staphylococcus epidermidis ; Streptococcus pneumoniae

Recently, there has been a growing demand for natural preservatives because of increased consumer interest in health. In this study, we produced Lactobacillus rhamnosus cell-free supernatant (LCFS) and evaluated and compared its antimicrobial activity with existing natural preservatives against pathogenic microorganisms and in chicken breast meat contaminated with Escherichia coli and Staphylococcus aureus. Lactobacillus rhamnosus cell-free supernatant possessed 30 units of lysozyme activity and contained 18,835 mg/L of lactic acid, 2,051 mg/L of citric acid and 5,060 mg/L of acetic acid. Additionally, LCFS inhibited the growth of fourteen pathogenic bacteria, S. aureus, Bacillus cereus, Listeria monocytogenes, Vibrio parahaemolyticus, Listeria innocua, S. epidermidis, L. ivanovii, E. coli, Pseudomonas aeruginosa, Shigella sonnei, Shi. flexneri, Proteus vulgaris, Pseudomonas fluorescens, and Klebsiella pneumoniae. The antibacterial activity of LCFS was stronger than that of egg white lysozyme (EWL), Durafresh (DF) and grapefruit seed extract (GSE). Additionally, LCFS maintained its antimicrobial activity after heat treatment at 50℃~95℃ and at pH values of 3~9. Moreover, LCFS inhibited the growth of E. coli and S. aureus in chicken breast meat. In conclusion, it is expected that LCFS, which contains both lysozyme and three organic acids, will be useful as a good natural preservative in the food industry.
Acetic Acid ; Bacillus cereus ; Bacteria ; Breast ; Chickens ; Citric Acid ; Citrus paradisi ; Egg White ; Escherichia coli ; Food Industry ; Hot Temperature ; Hydrogen-Ion Concentration ; Klebsiella pneumoniae ; Lactic Acid ; Lactobacillus rhamnosus ; Lactobacillus ; Listeria ; Listeria monocytogenes ; Meat ; Muramidase ; Proteus vulgaris ; Pseudomonas aeruginosa ; Pseudomonas fluorescens ; Shigella sonnei ; Staphylococcus aureus ; Vibrio parahaemolyticus

Vibrio vulnificus, a halophilic vibrio is an estuarine gram-negative bacteria that is associated with severe and frequently fatal wound infections and life-threatening septicemia. Bacteriocins are defined as antibacterial substance produced by various species of bacteria which are usually active against closely related organisms. Bacteriocins have found widespread application in epidemiological studies as specific markers of bacteria. It was proposed by Ha et al. (1990. J. Korean. Soc. Microbiol. 25: 586.) to give the bacteriocins produced by V. vulnificus the name "vulnificins". In the present study, a total of 72 strains of V. vulnificus isolated from patients and oysters were subjected to screen potential producers and indicators of vulnificin, applying ultraviolet induction method. Sensitivity of several strains of Serratia marcesans, Pseudomonas aeruginosa, Shigella flexneri, Salmonella typhi and Yersinia enterocolitica to vulnificins were also examined out. All the tested strains of V. vulnificus produced vulnificins active against indicator strains with various different inhibitory patterns. The spectrum of vulnificin activity and sensitive spectrum of indicator strains were considerably broad. Interestingly, almost all strains of S. marcescens, P. aeruginosa, Salmonella sp., Shigella sp. and Y. enterocolitica tested were sensitive to 1-7 vulnificin(s). Taken together, the present study demonstrated that all of the isolates of V. vulnificus produced vulnificins and that 8 good vulnificin producers and 10 good indicators were detected. These strains can be employed efficiently for establishing vulnificin typing scheme of V. vulnificus and for the detection of bacteriocinogeny and sensitivity in V. vulnificus. Biological role of vulnificin remains to be further elucidated.
Bacteria ; Bacteriocins ; Gram-Negative Bacteria ; Humans ; Ostreidae ; Pseudomonas aeruginosa ; Salmonella ; Salmonella typhi ; Sepsis ; Serratia ; Shigella ; Shigella flexneri ; Vibrio vulnificus* ; Vibrio* ; Wound Infection ; Yersinia enterocolitica

BACKGROUND: Recent clinical isolates of bacteria often produce various beta-lactamases, for example, extended spectrum beta-lactamase (ESBL) by some species of Enterobacteriaceae, TEM beta-lactamase by Haemophilus influenzae, and penicillinase by methicillin-susceptible Staphylococcus aureus (MSSA). Cefminox, a commonly used cephamycin, is stable to various beta-lactamases, but its activity against recent clinical isolates has not been evaluated. The aim of this study was to determine the activities of cefminox against recent clinical isolates of Enterobacteriaceae, H. influenzae, and MSSA. METHODS: The organisms, isolated from Severance Hospital patients during 1997 to 1998, were kept frozen until the test. Antimicrobial susceptibility was determined by the NCCLS agar dilution method. RESULTS: All 30 isolates of Escherichia coli and 90% of 30 Klebsiella pneumoniae isolates were susceptible to cefminox, cefotetan and amikacin. All of the Enterobacter cloacae and Serratia marcescens, and 86% of Citrobacter freundii isolates were susceptible to amikacin. All of the 15 isolates each of Proteus mirabilis, P. vulgaris and Morganella morganii were susceptible to cefminox, cefotetan, cefotaxime, ceftazidime, and aztreonam, while all of the Providencia spp. were susceptible to ceftazidime and aztreonam. All of the 29 H. influenzae isolates were susceptible to cefminox, cefotaxime, and levofloxacin, while all MSSA isolates were susceptible to cefoxitin and cotrimoxazole. CONCLUSION: Cefminox is more active than the other beta-lactams against Enterobacteriaceae including, ESBL- producing E. coli, and K. pneumoniae.
Agar ; Amikacin ; Aztreonam ; Bacteria ; beta-Lactamases ; beta-Lactams ; Cefotaxime ; Cefotetan ; Cefoxitin ; Ceftazidime ; Citrobacter freundii ; Enterobacter cloacae ; Enterobacteriaceae* ; Escherichia coli ; Haemophilus influenzae* ; Haemophilus* ; Humans ; Influenza, Human ; Klebsiella pneumoniae ; Levofloxacin ; Morganella morganii ; Penicillinase ; Pneumonia ; Proteus mirabilis ; Providencia ; Serratia marcescens ; Staphylococcus aureus* ; Staphylococcus* ; Trimethoprim, Sulfamethoxazole Drug Combination

BACKGROUND: In 2010, the Clinical and Laboratory Standards Institute (CLSI) revised breakpoints for cephalosporins and carbapenems and indicated that extended-spectrum beta-lactamase (ESBL) testing is no longer necessary for Enterobacteriaceae. We compared the results of the CLSI 2010 and the European Committee on Antimicrobial Susceptibility Testing (EUCAST) MIC breakpoints for Enterobacteriaceae producing ESBL and/or plasmid-mediated AmpC beta-lactamase (PABL). METHODS: A total of 94 well-characterized clinical isolates of Escherichia coli, Klebsiella oxytoca, Klebsiella pneumoniae, Proteus mirabilis, Salmonella spp., Shigella spp., Citrobacter freundii, Enterobacter aerogenes, Enterobacter cloacae, and Serratia marcescens were analyzed. Of them, 57 were ESBL producers, 24 were PABL producers, and 13 were ESBL plus PABL co-producers. Broth microdilution MIC tests were performed for cefotaxime, ceftazidime, aztreonam, cefepime, and imipenem. RESULTS: Among the 94 isolates containing ESBL and/or PABL, the number of isolates that were susceptible to cefotaxime, ceftazidime, aztreonam, cefepime, and imipenem according to the CLSI 2010 vs. the EUCAST breakpoints were 4 (4.3%) vs. 4 (4.3%); 26 (27.7%) vs. 8 (8.5%); 37 (39.4%) vs. 14 (14.9%); 71 (75.5%) vs. 31 (33.0%); and 76 (80.9%) vs. 90 (95.7%), respectively. Of the 18 isolates that were not susceptible to imipenem according to the CLSI 2010 breakpoints, 13 isolates (72.2%) were P. mirabilis. CONCLUSION: The CLSI 2010 MIC breakpoints without tests to detect ESBL and/or PABL for Enterobacteriaceae could be unreliable. Thus, special tests for ESBLs and AmpC beta-lactamases are required to detect the resistance mechanisms involved.
Aztreonam ; Bacterial Proteins ; beta-Lactamases ; beta-Lactams ; Carbapenems ; Cefotaxime ; Ceftazidime ; Cephalosporins ; Citrobacter freundii ; Enterobacter aerogenes ; Enterobacter cloacae ; Enterobacteriaceae ; Escherichia coli ; Imipenem ; Klebsiella oxytoca ; Klebsiella pneumoniae ; Proteus mirabilis ; Salmonella ; Serratia marcescens ; Shigella

To use in the identification of Legionella pneumophila isolates, one Legionella genus-specific, an two L. pneumophila species-specific monoclonal antibodies (MAbs) were produced. Reactivities of the MAbs with Legionella species and non-Legionella strains were tested by immunoblot. MAb 7D8-F1, MAb 7D8-A9, and MAb 1G12-C11 reacted only with protein. MAb 7D8-F1 recognized a genus-specific epitope in the protein ranged from 28 to 34 kDa in the tested 40 species with 61 serogroups. MAb 7D8-A9 and MAb 1G12-C11 reacted strongly with 49 kDa and 70 kDa protein from 18 L. pneumophila serogroups but did not react with 39 non-L. pneumophila species, respectively. In addtion, 17 non-Legionella strains, including Coxiella burnetii, Klebsiella pneumoniae, Mycoplasma pneumoniae, Pseudomonas areoginosa, Francisella tularensis, and Staphylococcus pneumoniae did not show cross-reacivities with the three MAbs. The MAbs reacted with 28 environmemtal isolates and three clinical isolates previously identified as L. pneumophila. When the immunoblot patterns were divided into four types (type I~IV) by using MAb 7D8-F1, all the 31 isolates belonged to the type II. These results indicate that the MAbs were highly specific to Legionella and can be used for the identification of L. pneumophila isolates on the genus and species levels.
Antibodies, Monoclonal* ; Coxiella burnetii ; Francisella tularensis ; Klebsiella pneumoniae ; Legionella pneumophila ; Legionella* ; Mycoplasma pneumoniae ; Pneumonia ; Pneumonia, Mycoplasma ; Pseudomonas ; Staphylococcus

BACKGROUND: There have been few studies about the kinds of species causing surgical site infections and their resistance pattern in Korea. An increase of extended-spectrum beta-lactamase (ESBL) producing strains is a worldwide problem. However, there is not enough data on the prevalence of ESBL-producing strains in Korea and the true extent of this problem seems to be under-recognized. METHODS: Minimal inhibitory concentrations of gram-negative bacilli isolated from surgical site infections were tested using the standard agar dilution method according to the National Committee for Clinical Laboratory Standards. To identify and characterize beta-lactamases, we performed conjugation test, isoelectric focusing, Southern hybridization, and polymerase chain reaction. RESULTS: A total of 54 strains of gram-negative enteric bacilli were identified:two strains of Acinetobacter spp., one of Citrobacter freundii, nine of Enterobacter cloacae, one of Enterobacter sakazakii, one of Escherichia coli, two of Klebsiella pneumoniae, one of Morganella morganii, one of Proteus vulgaris, 23 of Pseudomonas aeruginosa, four of Xanthomonas maltophila, and nine of Serratia marcescens. Three strains produced ESBL. CONCLUSION: Various species of gram-negative organisms isolated from surgical site infections showed complex antibiograms to various beta-lactams, even to the new generation of antibiotics. A large proportion of these strains showed conjugally transferable, plasmid-mediated, beta-lactam resistance. Some strains were ESBL-producing. This evidence suggests that there has been a molecular evolution of beta-lactamase genes to a great extent in Korea, possibly due to indiscriminate use of antibiotics.
Acinetobacter ; Agar ; Anti-Bacterial Agents ; beta-Lactam Resistance* ; beta-Lactamases ; beta-Lactams ; Citrobacter freundii ; Cronobacter sakazakii ; Drug Resistance ; Enterobacter cloacae ; Escherichia coli ; Evolution, Molecular ; Isoelectric Focusing ; Klebsiella pneumoniae ; Korea ; Microbial Sensitivity Tests ; Morganella morganii ; Polymerase Chain Reaction ; Prevalence ; Proteus vulgaris ; Pseudomonas aeruginosa ; Serratia marcescens ; Xanthomonas

BACKGROUND: There have been few studies about the kinds of species causing surgical site infections and their resistance pattern in Korea. An increase of extended-spectrum beta-lactamase (ESBL) producing strains is a worldwide problem. However, there is not enough data on the prevalence of ESBL-producing strains in Korea and the true extent of this problem seems to be under-recognized. METHODS: Minimal inhibitory concentrations of gram-negative bacilli isolated from surgical site infections were tested using the standard agar dilution method according to the National Committee for Clinical Laboratory Standards. To identify and characterize beta-lactamases, we performed conjugation test, isoelectric focusing, Southern hybridization, and polymerase chain reaction. RESULTS: A total of 54 strains of gram-negative enteric bacilli were identified:two strains of Acinetobacter spp., one of Citrobacter freundii, nine of Enterobacter cloacae, one of Enterobacter sakazakii, one of Escherichia coli, two of Klebsiella pneumoniae, one of Morganella morganii, one of Proteus vulgaris, 23 of Pseudomonas aeruginosa, four of Xanthomonas maltophila, and nine of Serratia marcescens. Three strains produced ESBL. CONCLUSION: Various species of gram-negative organisms isolated from surgical site infections showed complex antibiograms to various beta-lactams, even to the new generation of antibiotics. A large proportion of these strains showed conjugally transferable, plasmid-mediated, beta-lactam resistance. Some strains were ESBL-producing. This evidence suggests that there has been a molecular evolution of beta-lactamase genes to a great extent in Korea, possibly due to indiscriminate use of antibiotics.
Acinetobacter ; Agar ; Anti-Bacterial Agents ; beta-Lactam Resistance* ; beta-Lactamases ; beta-Lactams ; Citrobacter freundii ; Cronobacter sakazakii ; Drug Resistance ; Enterobacter cloacae ; Escherichia coli ; Evolution, Molecular ; Isoelectric Focusing ; Klebsiella pneumoniae ; Korea ; Microbial Sensitivity Tests ; Morganella morganii ; Polymerase Chain Reaction ; Prevalence ; Proteus vulgaris ; Pseudomonas aeruginosa ; Serratia marcescens ; Xanthomonas

BACKGROUND: We compared the performance of the modified Hodge test (MHT), Triton Hodge test (THT), Carba NP test (CNPt), simplified Carba NP test (CNPt-direct), blue-Carba NP test (BCT), and carbapenem inactivation method (CIM) for rapid and accurate carbapenemase detection. METHODS: The methods were evaluated by using 256 gram-negative isolates, including 197 Enterobacteriaceae (79 Enterobacter spp., 74 Klebsiella spp., 33 Escherichia coli, 10 Citrobacter spp., and 1 Serratia marcescens), 51 Acinetobacter baumannii, and 8 Pseudomonas aeruginosa strains. The collection included 117 non-carbapenemase, 18 Klebsiella pneumoniae carbapenemases (KPC) producers, 46 New Delhi metallo-β-lactamases (NDM) producers, 11 imipenemases (IMP) producers, and 51 oxacillinases (OXA) producers, and 13 strains harboring two different carbapenemase genes. RESULTS: The specificity of the THT (91.5%) was significantly lower than other methods, each of which had 100% specificity (P<0.003). This can be attributed to the false detection of Ampler class C β-lactamases (AmpC) carriers. The CNPt-direct and CIM yielded the highest sensitivities (P<0.003), which were comparable (92.8% vs 93.5%, P>0.999). Because of improved detection of NDM carriers, THT showed significantly higher sensitivity than the MHT (84.9% vs 75.5%, P<0.001). However, poor performances in detecting OXA still influenced the sensitivities of the CNPt (66.2%) and BCT (82.0%), as well as the MHT and THT. CONCLUSIONS: CNPt-direct and CIM demonstrated the best performance for the efficient detection of carbapenemase among the six evaluated methods. Except the MHT and THT, the detection of carbapenemase-producing Enterobacteriaceae by all the other methods was acceptable, when the OXA-type carbapenemase was not prevalent.
Acinetobacter baumannii ; Citrobacter ; Enterobacter ; Enterobacteriaceae ; Escherichia coli ; Gram-Negative Bacteria* ; Klebsiella ; Klebsiella pneumoniae ; Methods* ; Neptune ; Pseudomonas aeruginosa ; Sensitivity and Specificity ; Serratia