This study was performed to investigate polymerase chain reaction-based detection of bacterial DNA in middle ear fluid and assess the correlation between the PCR-positive rate with several factors associated with middle ear effusion. The purpose was to gain a further understanding of bacterial infection as a major cause of otitis media with effusion. Of the 278 specimens of middle ear fluid, 39 (14%) tested positive by ordinary culture. The overall detection rate of bacterial DNA using the PCR method was 36.7% for middle ear effusion, and bacterial DNA detection rates of Hemophilus influenzae, Streptococcus pneumoniae, Moraxella catarrhalis in the middle ear effusion were 29.1%, 4.7% and 10.8%, respectively. The bacterial DNA detection rate was higher in ears with a history of acute otitis media than those without the history. High detection rates were observed in patients younger than 48 months who have had a higher tendency to present with acute otitis media. We concluded that PCR is a more sensitive method for the detection of bacteria in middle ear effusion than ordinary culture, and acute otitis media is a major contributor to the pathogenesis of otitis media with effusion.
Child ; Child, Preschool ; Chronic Disease ; DNA, Bacterial/analysis ; Haemophilus Infections/*diagnosis ; Haemophilus influenzae/genetics/*isolation & purification ; Humans ; Infant ; Moraxella (Branhamella) catarrhalis/genetics/isolation & purification ; Moraxellaceae Infections/diagnosis ; Otitis Media with Effusion/*diagnosis/*microbiology ; Polymerase Chain Reaction ; Research Support, Non-U.S. Gov't ; Streptococcal Infections/diagnosis ; Streptococcus pneumoniae/genetics/isolation & purification

We had three cases of Moraxella osloensis meningitis. The species identification was impossible by conventional and commercial phenotypic tests. However, we could identify the species using the 16S rRNA gene sequencing. Determination of clinical significance was difficult in one patient. All three patients recovered by appropriate antimicrobial therapy.
Adolescent ; Aged, 80 and over ; Anti-Bacterial Agents/therapeutic use ; *Bacterial Typing Techniques ; Child, Preschool ; Female ; Humans ; Male ; Meningitis, Bacterial/drug therapy/*microbiology ; Moraxella/*pathogenicity ; Moraxellaceae Infections/drug therapy/*microbiology ; RNA, Bacterial/analysis ; RNA, Ribosomal, 16S/analysis

Members of the genus Acinetobacter are recognized as newer pathogens of the nosocomial infection with an increasing frequency in recent years. Strains that belonged to A. calcoaceticus-A. baumannii complex (genomic species 1, 2, 3, and 13TU) were major groups associated with nosocomial infection. Phenotypic identification was unreliable and laborious method to classify Acinetobacter strains into 19 genomic species. Rapid and reliable identification of clinical isolates is essential to diagnosis and epidemiology of Acinetobacter. We investigated the suitability of amplified ribosomal DNA restriction analysis (ARDRA) to identify genomic species of 131 Acinetobacter isolates. The 16S rRNA genes (ribosomal DNA) were enzymatically amplified and the amplified PCR products were restricted independently with the enzymes, AluI, CfoI, and MboI. Genomic species of Acinetobacter was classified by the combinations of restriction patterns. The analysis was showed that restriction profiles were characteristic for each genomic species. One hundred fourteen isolates were identified as A. baumannii, twelve were identified as genomic species 13TU, and one was identified as genomic species 3. Four isolates were found to be unknown organisms. All of the isolates which were identified to A. baumannii by phenotypic tests were completely discriminated into A. baumannii and genomic species 13TU by ARDRA. This study demonstrates that ARDRA is a rapid and simple techniques for the identification of Acinetobacter species according to the genomic species.
Acinetobacter baumannii* ; Acinetobacter calcoaceticus* ; Acinetobacter* ; Cross Infection ; Diagnosis ; DNA, Ribosomal* ; Epidemiology ; Genes, rRNA ; Polymerase Chain Reaction

Acinetobacter species encounters frequently with clinical specimens and now accounts for a substantial proportion of endemic nosocomial infections in Korea. Recent trends indicate that the antimicrobial resistant strains of Acinetobacter species are increasing. Sixty-one strains were isolated from specimens of patients suspected of nosocomial infections during 1991 to 1996. At present, phenotypic identification of Acinetobacter using biochemical test may not be reliable and resulted in the difficulty to clarify the source of infections and epidemiological study of hospital-acquired infections. Aware of the importance of rational taxonomic proposal for these isolates, correct species identification of these organisms by molecular typing method was carried out. A total of fifty-four strains of A. calcoaceticus-A. baumannii complex species which were identified to genospecies 2 and 13 by biochemical characteristics was subjected to identify by ribotyping using restriction endonuclease EcoRI, ClaI, and SalI. Of fifty-four strains, twenty-five strains were identified as A. baumannii (genospecies 2) and twenty-one strains as genospecies 13, and six strains changed to genospecies 3, and the rest two strains were confirmed as A. haemolyticus (genospecies 4). This result suggests that the ribotyping may be of value for identification of genospecies and epidemiological information of Acinetobacter strains.
Acinetobacter baumannii* ; Acinetobacter calcoaceticus* ; Acinetobacter* ; Cross Infection ; DNA Restriction Enzymes ; Humans ; Korea ; Molecular Typing ; Ribotyping*

No abstract available.
Acinetobacter calcoaceticus* ; Acinetobacter* ; Pneumonia*

No abstract available.
Aged ; Ampicillin/therapeutic use ; Anti-Bacterial Agents/therapeutic use ; Antimetabolites, Antineoplastic/therapeutic use ; Asian Continental Ancestry Group ; Cytarabine/therapeutic use ; Drug Therapy, Combination ; Humans ; Idarubicin/therapeutic use ; Leukemia, Myeloid, Acute/complications/*diagnosis/drug therapy ; Male ; Moraxella/genetics/*isolation & purification ; Moraxellaceae Infections/*diagnosis/drug therapy/microbiology ; RNA, Ribosomal, 16S/chemistry/genetics ; Republic of Korea ; Respiratory Tract Infections/diagnosis/microbiology ; Sequence Analysis, RNA ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Sulbactam/therapeutic use

OBJECTIVEGas chromatography (GC) was used to investigate the cellular fatty acid (CFA) composition of 141 Acinetobacter baumannii and 32 A. calcoaceticus isolates from different locations in China and to find chemical markers to differentiate these two closely related bacteria.

METHODSWhole cell fatty acid methyl esters (FAMEs) were obtained by saponification, methylation, and extraction for GC analysis, followed by a standardized Microbial Identification System (MIS) analysis.

RESULTSAll A. baumannii and A. calcoaceticus strains contained some major fatty acids, namely, 18:1 ω9c, 16:0, Sum In Feature 3, 12:0, 17:1ω8c, 3-OH-12:0, 17:0, Sum In Feature 2, 2-OH-12:0, and 18:0 compounds. Although most of the total CFAs are similar between A. baumannii and A. calcoaceticus strains, the ratios of two pairs of CFAs, i.e., Sum In Feature 3/18:1 ω9c versus 16:0/18:1 ω9c and Sum In Feature 3/18:1 ω9c versus unknown 12.484/18:1 ω9c fatty acids, could differentiate these two closely related bacteria. A. baumannii could be easily classified into two subgroups by plotting some ratios such as Sum In Feature 3/16:0 versus 17:0 and Sum In Feature 3/2-OH-12:0 versus 17:0 fatty acids.

CONCLUSIONThe ratios of some CFAs could be used as chemical markers to distinguish A. baumannii from A. calcoaceticus.

Acinetobacter baumannii ; classification ; cytology ; metabolism ; Acinetobacter calcoaceticus ; classification ; cytology ; metabolism ; Biomarkers ; metabolism ; Fatty Acids ; metabolism ; Species Specificity

BACKGROUND: Animal skin provides an ideal medium for the propagation of microorganisms and it is used like raw material in the tannery and footware industry. The aim of this study was to evaluate and identify the microbial load in oropharyngeal mucosa of tannery employees. METHODS: The health risk was estimated based on the identification of microorganisms found in the oropharyngeal mucosa samples. The study was conducted in a tanners group and a control group. Samples were taken from oropharyngeal mucosa and inoculated on plates with selective medium. In the samples, bacteria were identified by 16S ribosomal DNA analysis and the yeasts through a presumptive method. In addition, the sensitivity of these microorganisms to antibiotics/antifungals was evaluated. RESULTS: The identified bacteria belonged to the families Enterobacteriaceae, Pseudomonadaceae, Neisseriaceae, Alcaligenaceae, Moraxellaceae, and Xanthomonadaceae, of which some species are considered as pathogenic or opportunistic microorganisms; these bacteria were not present in the control group. Forty-two percent of bacteria identified in the tanners group are correlated with respiratory diseases. Yeasts were also identified, including the following species: Candida glabrata, Candida tropicalis, Candida albicans, and Candida krusei. Regarding the sensitivity test of bacteria identified in the tanners group, 90% showed sensitivity to piperacillin/tazobactam, 87% showed sensitivity to ticarcillin/clavulanic acid, 74% showed sensitivity to ampicillin/sulbactam, and 58% showed sensitivity to amoxicillin/clavulanic acid. CONCLUSION: Several of the bacteria and yeast identified in the oropharyngeal mucosa of tanners have been correlated with infections in humans and have already been reported as airborne microorganisms in this working environment, representing a health risk for workers.
Alcaligenaceae ; Animals ; Bacteria ; Candida ; Candida albicans ; Candida glabrata ; Candida tropicalis ; DNA, Ribosomal ; Enterobacteriaceae ; Humans ; Moraxellaceae ; Mucous Membrane* ; Neisseriaceae ; Pseudomonadaceae ; Skin ; Xanthomonadaceae ; Yeasts

BACKGROUND: Cefditoren is an oral cephalosporin with excellent activity against Haemophilus influenzae, Streptococcus pneumoniae, and Moraxella catarrhalis, which are the predominant bacterial causes of community-acquired respiratory tract infections. The current study attempted to determine the antibacterial activity of cefditoren against the major clinical isolates. METHODS: According to the NCCLS recommendations, antibacterial activities of cefditoren were measured against total 504 major clinical isolates. MICs were determined by the agar dilution method, a series of doubling dilutions from 128 to 0.03 microgram/mL, on E. coli, K. pneumoniae, E. cloacae, C. freundii, S. marcescens, P. mirabilis, and Staphylococcus spp. In case of H. influenzae, S. pneumoniae, and M. catarrhalis, broth microdilution method, a series of doubling dilutions from 16 to 0.015 microgram/mL, was performed. RESULTS: Cefditoren had variable activity against Enterobacteriaceae. MIC cumulative curves showed that cefditoren had low MIC distributions against E. coli and P. mirabilis, and MIC90 were 8 and 0.5 microgram/mL, respectively. Against K. pneumoniae, E. cloacae, C. freundii, and S. marcescens, cefditoren's MIC90 values ranged from 32 to >128 microgram/mL. For clinical isolates of methicillin-susceptible S. aureus and methicillin-susceptible S. epidermidis, cefditoren had MIC90 values of 1 microgram/mL and 0.5 microgram/mL, respectively. Cefditoren had MIC90 values of 1 microgram/mL for penicillin-susceptible and penicillin-not-susceptible strains of S. pneumoniae. Cefditoren had MIC90 values of 0.03 microgram/mL and 0.5microgram/mL against H. influenzae and M. catarrhalis, respectively. CONCLUSION: Cefditoren had excellent activity against S. pneumoniae, H. influenzae, and M. catarrhalis. Cefditoren had variable activity against Enterobacteriaceae. The results of this study confirm the excellent activity of cefditoren against the major respiratory tract pathogens and suggest that cefditoren could be a good antibiotic for empiric oral treatment of community-acquired respiratory tract infections.
Agar ; Cloaca ; Enterobacteriaceae ; Haemophilus influenzae ; Influenza, Human ; Mirabilis ; Moraxella (Branhamella) catarrhalis ; Pneumonia ; Respiratory System ; Respiratory Tract Infections ; Staphylococcus ; Streptococcus pneumoniae

BACKGROUND: Cefditoren is an oral cephalosporin with excellent activity against Haemophilus influenzae, Streptococcus pneumoniae, and Moraxella catarrhalis, which are the predominant bacterial causes of community-acquired respiratory tract infections. The current study attempted to determine the antibacterial activity of cefditoren against the major clinical isolates. METHODS: According to the NCCLS recommendations, antibacterial activities of cefditoren were measured against total 504 major clinical isolates. MICs were determined by the agar dilution method, a series of doubling dilutions from 128 to 0.03 microgram/mL, on E. coli, K. pneumoniae, E. cloacae, C. freundii, S. marcescens, P. mirabilis, and Staphylococcus spp. In case of H. influenzae, S. pneumoniae, and M. catarrhalis, broth microdilution method, a series of doubling dilutions from 16 to 0.015 microgram/mL, was performed. RESULTS: Cefditoren had variable activity against Enterobacteriaceae. MIC cumulative curves showed that cefditoren had low MIC distributions against E. coli and P. mirabilis, and MIC90 were 8 and 0.5 microgram/mL, respectively. Against K. pneumoniae, E. cloacae, C. freundii, and S. marcescens, cefditoren's MIC90 values ranged from 32 to >128 microgram/mL. For clinical isolates of methicillin-susceptible S. aureus and methicillin-susceptible S. epidermidis, cefditoren had MIC90 values of 1 microgram/mL and 0.5 microgram/mL, respectively. Cefditoren had MIC90 values of 1 microgram/mL for penicillin-susceptible and penicillin-not-susceptible strains of S. pneumoniae. Cefditoren had MIC90 values of 0.03 microgram/mL and 0.5microgram/mL against H. influenzae and M. catarrhalis, respectively. CONCLUSION: Cefditoren had excellent activity against S. pneumoniae, H. influenzae, and M. catarrhalis. Cefditoren had variable activity against Enterobacteriaceae. The results of this study confirm the excellent activity of cefditoren against the major respiratory tract pathogens and suggest that cefditoren could be a good antibiotic for empiric oral treatment of community-acquired respiratory tract infections.
Agar ; Cloaca ; Enterobacteriaceae ; Haemophilus influenzae ; Influenza, Human ; Mirabilis ; Moraxella (Branhamella) catarrhalis ; Pneumonia ; Respiratory System ; Respiratory Tract Infections ; Staphylococcus ; Streptococcus pneumoniae