The purpose of this study is to investigate the correlation of cell surface hydrophobicity (CSH) and biofilm formation or adhesion in Candida albicans (C. albicans) and several pathogenic bacteria. All of C. albicans (n=82) and 7 bacterial species (Escherichia coli, n=25; Klebsiella pneumoniae, n=33; Morganella morganii, n=21; Proteus mirabilis, n=33; Proteus vulgaris, n=12; Pseudomonas aeruginosa, n=31; Staphylococcus aureus, n=31) were isolated clinically. CSH was quantified with microbial adhesion to hydrocarbons. Biofilm formation was determined by tetrazolium salt reduction assay. Adhesion assay was performed by counting colonies after culture the microbes adhered to HeLa cells. Although high CSH-expressing bacterial species showed greater adherence to HeLa cells and larger amounts of biofilm formation on polystyrene, the significant relationships within same species were not shown. In C. albicans, however, strong positive correlations were observed between CSH and biofilm formation (r =0.708; p < 0.05) or cell adhesion (r =0.509; p < 0.05). These results suggest that hydrophobic force of bacteria may play a minor role in adhesion and biofilm formation, but CSH of C. albicans may be an important factor for adherence on surface and biofilm forming process.
Bacteria ; Biofilms* ; Candida albicans* ; Candida* ; Cell Adhesion ; HeLa Cells ; Humans ; Hydrocarbons ; Hydrophobic and Hydrophilic Interactions* ; Klebsiella pneumoniae ; Morganella morganii ; Polystyrenes ; Proteus mirabilis ; Proteus vulgaris ; Pseudomonas aeruginosa ; Staphylococcus aureus

We evaluated the impact of revised Clinical and Laboratory Standards Institute (CLSI) breakpoints for broad-spectrum cephalosporins (BSCs) on the susceptibilities of 1,742 isolates of Enterobacter species, Serratia marcescens, Citrobacter freundii, and Morganella morganii. The 2011 CLSI criteria for cefotaxime and ceftazidime reduced the rates of susceptibility by 2.9% and 5.9%, respectively. The 2014 CLSI criteria for cefepime reduced the rate of susceptibility by 13.9%, and categorized 11.8% isolates as susceptible-dose dependent (SDD) for cefepime. Among 183 isolates with extended-spectrum ß-lactamase (ESBL) phenotype, implementation of the new criteria reduced the rates of susceptibility to cefotaxime, ceftazidime, and cefepime by 2.8%, 14.8%, and 53.6%, respectively. The proportion of ESBL phenotype among BSC-susceptible isolates was low (0.9% for cefotaxime, 3.0% for ceftazidime, and 3.3% for cefepime). In summary, implementation of new CLSI criteria led to little change in susceptibility to cefotaxime and ceftazidime but a substantial change in susceptibility to cefepime. The recognition of revised CLSI criteria for BSC and SDD will help clinicians to select the optimal antibiotic and dosing regimen.
Cefotaxime ; Ceftazidime ; Cephalosporins ; Citrobacter freundii ; Enterobacter ; Enterobacteriaceae* ; Morganella morganii ; Phenotype ; Serratia marcescens

Enantioselective hydrolysis of 2-carboxyethyl-3-cyano-5-methylhexanoic acid (CNDE) is the key step in chemoenzymatic synthesis of pregabalin. We purified an intracellular carboxyl esterase from Morganella morganii ZJB-09203, which exhibited high enantioselectivity and activity towards CNDE. The carboxyl esterase was purified to electrophoretic homogeneity by ammonium sulfate fraction precipitation, Phenyl Sepharose 6 FF hydrophobic interaction chromatography, anion exchange with DEAE Sephadex A-50 and Bio-Scale CHT column. The purified enzyme was a monomer with molecular mass of 68 kDa determined by SDS-PAGE and gel chromatography. Substrate specificity of the enzyme towards p-nitrophenyl esters suggested that the purified enzyme was an esterase. The optimal reaction pH for CNDE hydrolysis was 9.0, and optimal temperature was 45 degrees C. The esterase was stable between pH 7.0 and 9.0, and at 40 degrees C. The enzyme activity was enhanced by Ca2+, Cu2+ and Mn2+, whereas strongly inhibited by Co2+, Fe3+, Ni2+ and EDTA. Meanwhile, we investigated the kinetic parameters of the esterase towards p-nitrophenyl esters and effect of CNDE concentration on conversion. The present study reported the esterase capable of stereospecific hydrolysis of CNDE for the first time. Our research will provide foundations for industrial production of Pregabalin using the new biocatalyst.
Chromatography, Gel ; Electrophoresis, Polyacrylamide Gel ; Esterases ; metabolism ; Hydrogen-Ion Concentration ; Kinetics ; Molecular Weight ; Morganella morganii ; enzymology ; Substrate Specificity ; Temperature

PURPOSE: To report a case of spontaneous eye ball rupture without trauma in a 94-year-old patient. CASE SUMMARY: A 94-year-old female patient diagnosed with cataract in both eyes 20 years was referred to this ophthalmologic department for treatment consultation of a painful left eye with spontaneous bleeding. She has used anti-cataract eye drops and artificial tears three times a day for several years without consulting a doctor. Fifteen days prior to presentation, the patient suffered severe left eyeball pain and headache and was diagnosed with acute angle-closure glaucoma secondary to hypermature cataract. She underwnet eviceration after ocular examination and systemic evaluation. Surgical findings included a thin cornea at the inferior limbus and protruding intraocular tissues. Additionally, the eyeball was filled with a blood clot from a choroidal hemorrhage. Morganella morganii were grown in a bacterial swap culture, and a corneal biopsy revealed suppurative inflammation. CONCLUSIONS: In old age, a thin corneal limbus due to infection and complicated acute angle-closure glaucoma can cause massive suprachoroidal hemorrhage with spontaneous eyeball rupture.
Biopsy ; Cataract ; Choroid Hemorrhage ; Cornea ; Eye ; Female ; Glaucoma ; Glaucoma, Angle-Closure ; Headache ; Hemorrhage ; Humans ; Limbus Corneae ; Morganella morganii ; Ophthalmic Solutions ; Rupture

BACKGROUND: Among the inducible AmpC beta-lactamase-producing members of the family Enterobacteriaceae such as Enterobacter spp., Citrobacter spp., Serratia spp., and Morganella morganii (ECSM), the prevalence of ESBL-producing isolates are increasing. However, there have been only a limited number of studies that have investigated the prevalence for ESBL-production in blood isolates of these organisms. MATERIALS AND METHODS: We performed a prospective observational study to evaluate the prevalence for ESBL production among ECSM blood isolates. All consecutive blood isolates in the Samsung Medical Center were included from Oct 2006 to Mar 2008. Antimicrobial susceptibility test was performed by broth microdilution method. ESBLs were confirmed by double-disk synergy test and ESBL phenotypes were determined by PCR. RESULTS: The 124 isolates (94 Enterobacter spp., 18 Citrobacter spp., 8 Serratia spp. and 4 Morganella spp.) were investigated. Among 124 ESCM isolates, 30 isolates (24.2%) showed ESBL-producing activity. Derepressed or partially derepressed AmpC mutants and derepressed AmpC mutants with ESBL production accounted for 36.3% (45/124) and 16.9% (21/124), respectively. Of ESBL producers, the most prevalent ESBL was SHV-12 (5/24, 20.8%). CONCLUSIONS: The prevalence of ESBL-producing isolates is high in Enterobacter spp., Serratia marcescens and Citrobacter spp. clinical isolates. It suggested that routine screening test for ESBLs among Enterobacteriacae blood isolates with inducible AmpC beta-lactamase should be needed.
Bacterial Proteins ; beta-Lactamases ; Citrobacter ; Enterobacter ; Enterobacteriaceae ; Humans ; Mass Screening ; Morganella ; Morganella morganii ; Phenotype ; Polymerase Chain Reaction ; Prevalence ; Prospective Studies ; Serratia ; Serratia marcescens

BACKGROUND: Clinical isolates of AmpC beta-lactamase- producing Enterobacteriaceae were evaluated to determine the prevalence of CTX-M extended-spectrum beta-lactamases (ESBLs) and their genetic environments. METHODS: A total of 250 non-duplicate isolates of Eneterobacter aerogenes, E. cloacae, Citrobacter freundii, Serratia marcescens and Morganella morganii were collected at a Korean hospital. ESBL production was determined by double disk synergy test. For ESBL producers, bla genes were sequenced and blaCTX-M environment was characterized by PCR mapping and sequencing. RESULTS: Among the 250 isolates 29 (11.6%) produced ESBL, and 14 of the 29 isolates produced CTX-M ESBLs, including CTX-M-9 by 8 isolates, CTX-M-3 by 4 isolates, CTX-M-12 by 1 isolate, and CTX-M-14 by 1 isolate. ISEcp1 was present upstream of blaCTX-M-3, 12, and 14. Three of the four CTX- M-3 producers had the same genetic environment (pemK-ISEcp1-blaCTX-M-3-orf477-mucA). An IS903-like element was found downstream of blaCTX-M-14. ISCR1 was identified upstream of blaCTX-M-9 and ISCR1 and blaCTX-M-9 were located on sul1-type class 1 integron. The variable region between the 5'-CS and the first 3'-CS contained dfrA16 and aadA2. Its structure was similar to that of In60, but our isolates did not have IS3000 or second 3'-CS. CONCLUSION: CXT-M type ESBL was prevalent in AmpC beta-lactamase-producing Enterobacteriaceae, particularly E. cloacae. blaCTX-M genes were associated with ISEcp1 or ISCR1. This is the first report on the genetic environment of blaCTX-M in Korean isolates.
beta-Lactamases ; Citrobacter freundii ; Cloaca ; Enterobacteriaceae ; Integrons ; Korea ; Morganella morganii ; Polymerase Chain Reaction ; Prevalence ; Serratia marcescens

The purification and the characteristics of an enzyme from Morganella morganii J-8, which could produce d-pseudoephedrine from 1-phenyl-2-methylamine-acetone, were performed in this study. In this research, first, cells were disrupted by ultrasonic treatment at 4 degrees C. The carbonyl enantioselective reductase was purified with a combination of ammonium precipitation, Phenyl Superose hydrophobic chromatography, DEAE anion exchange, and native polyacrylamide gel electrophoresis. The molecular mass of the purified enzyme subunit was estimated to be 42.5kD on sodium dodecyl sulfate-polyacrylamide electrophoresis (SDS-PAGE). The native molecular mass of the enzyme that was analyzed by high-performance liquid chromatography was found out to be 84.1 kD, which indicated that the enzyme was a dimmer. The purified enzyme was analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and the result showed that the purified enzyme had high homology with leucine dehydrogenase.
Bacterial Proteins ; chemistry ; isolation & purification ; metabolism ; Biocatalysis ; drug effects ; Chromatography, High Pressure Liquid ; Electrophoresis, Polyacrylamide Gel ; Enzyme Stability ; drug effects ; Hydrogen-Ion Concentration ; Kinetics ; Leucine Dehydrogenase ; metabolism ; Metals, Heavy ; pharmacology ; Molecular Weight ; Morganella morganii ; enzymology ; metabolism ; Oxidoreductases ; chemistry ; isolation & purification ; metabolism ; Pseudoephedrine ; chemistry ; metabolism ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Stereoisomerism ; Temperature

Morganella morganii is a facultative gram-negative and anaerobic rod. It may be a cause of devastating infections in neonates and immunocompromised hosts. Some bacterial infections such as Clostridium and Vibrio are associated with hemolysis. However, massive hemolysis caused by M. morganii sepsis has not yet been reported. We observed a 59-yr-old man who had chemotherapy-induced neutropenia and was found to have massive hemolysis and metabolic acidosis due to sepsis. He died 6 hr after admission in spite of aggressive treatment. Two sets of blood cultures revealed the growth of M. morganii. We report here that M. morganii sepsis can cause fatal massive hemolysis leading to death.
Antineoplastic Agents/adverse effects ; Bacteremia/*complications ; Enterobacteriaceae Infections/*complications ; *Hemolysis ; Humans ; Male ; Middle Aged ; *Morganella morganii ; Neutropenia/complications

PURPOSE: Bacteremia in immunocompromised pediatric cancer patients can lead to high morbidity and mortality, if not treated early and properly. The incidence and antibiotic sensitivities to common pathogens of bacteremia in pediatric cancer patients are liable to change, according to region and time. We investigated the causative organisms and antibiotic sensitivities of bacteremia in pediatric cancer patients to assess the adequacy of empiric antimicrobial therapy. METHODS: From September 1995 to August 2003, we retrospectively evaluated 58 episodes in 39 pediatric cancer patients with bacteremia treated at the Pediatric Department of Yeungnam University Hospital. We investigated and analyzed the causative organisms and the antibiotic sensitivity test results by reviewing the records of the microbiologically proven positive blood culture results. RESULTS: The incidence of bacteremia in pediatric cancer patients in this study was 5.7 percent (58 episodes out of 1, 022 occasions of blood cultures). Gram-positive organisms were isolated more often than gram-negative organisms (63.8 percent vs 36.2 percent) in the following order: Staphylococcus epidermidis (37.9 percent), Staphylococcus aureus (17.3 percent), Escherichia coli (12 percent), Streptococcus (8.6 percent), Enterobacter (6.9 percent), Klesiella (6.9 percent), Serratia (3.5 percent), Acinetobacter (3.5 percent), Proteus (1.7 percent) and Morganella morganii (1.7 percent). In antibiotic sensitivity tests, only six of 37 isolates (16 percent) of gram positive bacteria were sensitive to penicillin and 15 of 37 isolates (40 percent) were sensitive to oxacillin. All except one Staphylococcus aureus were sensitive to vancomycin and all except one Staphylococcus epidermidis were sensitive to teicoplanin among 37 isolates of gram positive bacteria. In the case of gram negative bacteria, two of 21 isolates (10 percent) and four of 21 isolates (19 percent) were sensitive to cefotaxime and ceftazidime, respectively. Only six of 21 isolates (29 percent) were sensitive to aminoglycoside, but all 21 isolates (100 percent) were sensitive to imipenem. All seven isolates tested after the year 2000 were sensitive to meropenem. CONCLUSION: In conclusion, we should choose the proper antimicrobials in treating pediatric cancer patients with suspected bacteremia, reflecting the increasing episodes of gram positive bacteremia and polymicrobial resistance of gram positive and negative organisms.
Acinetobacter ; Bacteremia* ; Cefotaxime ; Ceftazidime ; Enterobacter ; Escherichia coli ; Gram-Negative Bacteria ; Gram-Positive Bacteria ; Humans ; Imipenem ; Incidence ; Morganella morganii ; Mortality ; Oxacillin ; Penicillins ; Proteus ; Retrospective Studies ; Serratia ; Staphylococcus aureus ; Staphylococcus epidermidis ; Streptococcus ; Teicoplanin ; Vancomycin

PURPOSE: The main objectives of this work were to characterize the mechanism of resistance to imipenem of Morganella morganii KU158 isolated in Busan, Korea and to analyze the structure of the integron, which carries the resistance gene that confers resistance to imipenem. MATERIALS AND METHODS: Antimicrobial susceptibilities of M. morganii KU158 were tested by using the disk diffusion method. The modified Hodge and EDTA-disk synergy tests were performed for the screening of metallo-beta-lactamase-producing isolates. blaIMP and blaVIM genes were detected using the polymerase chain reaction (PCR) amplification. To detect the presence of the integron, the PCR method was used. The PCR product was cloned through the use of primers, 5'CS-F and 3'CS-R, and it was used to determine the sequence of the integron through the dideoxy-mediated chain termination method. RESULTS: M. morganii KU158 was intermediately resistant to imipenem and showed a positive result for the modified Hodge and EDTA-disk synergy tests, which suggest the production of metallo-beta-lactamase, and also was positive in the PCR result for the detection of blaVIM gene. The genotype of the PCR product from the blaVIM gene was blaVIM-2. Sequencing of the 5,031 bp-cloned fragment revealed the structure of the class I integron, such as the 5'-CS element containing an Intl1 integrase gene with its own promoter region, the attI1 recombination site, and the 3'-CS element containing qacE1. The integron contained insert gene cassettes blaVIM-2, aac(6')-Ib, aadA1, "orfII", and "orfIII". The blaVIM-2 gene was located immediately downstream of the aac(6')-Ib gene. CONCLUSIONS: M. morganii KU158 acquired the resistance to imipenem through the production of metallo-beta-lactamase VIM-2. The gradual increase in the number of VIM-2-producing bacterial species may indicate the highly mobile nature of the blaVIM-2 cassette. The spread of blaVIM-2 could compromise the future usefulness of carbapenem in treating gram-negative bacilli infections.
beta-Lactamases ; Busan ; Clone Cells ; Diffusion ; Genotype ; Imipenem ; Integrases ; Integrons* ; Korea ; Mass Screening ; Morganella morganii* ; Morganella* ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; Recombination, Genetic