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From December 2002 to February 2003, 15 isolates of pandrug-resistant unidentified Acinetobacter species were recovered from seven patients treated on different wards or intensive care units. Both 16S-23S rRNA intergenic spacer PCR-restriction fragment length polymorphism profiles and sequence analysis of these isolates identified them as Acinetobacter baumannii . This pandrug-resistant A. baumannii strain with an unusual phenotype could persist in humans for long periods and was widely disseminated throughout the hospital.

We evaluated the disk susceptibility data of 671 nontyphoid Salmonella isolates collected from different parts of Taiwan from March 2001 to August 2001 and 1,261 nontyphoid Salmonella isolates from the National Taiwan University Hospital from 1996 to 2001. Overall, ciprofloxacn resistance was found in 2.7% (18/671) of all nontyphoid Salmonella isolates, in 1.4% (5/347) of Salmonella enterica serotype Typhimurium and in 7.5% (8/107) in S. enterica serotype Choleraesuis nationwide. MICs of six newer fluoroquinolones were determined for the following isolates: 37 isolates of ciprofloxacin-resistant (human) S. enterica Typhimurium (N = 26) and Choleraesuis (N = 11), 10 isolates of ciprofloxacin-susceptible (MIC 0.12 μg/mL). Sequence analysis of the gryA, gyrB, parC, parE, and acrR genes, ciprofloxacin accumulation; and genotypes generated by pulsed-field gel electrophoresis with three restriction enzymes (SpeI, XbaI, and BlnI) were performed. All 26 S. enterica Typhimurium isolates from humans and pigs belonged to genotype I. For S. enterica Choleraesuis isolates, 91% (10/11) of human isolates and 54% (7/13) of swine isolates belonged to genotype B. These two genotypes isolates from humans all exhibited a high-level of resistance to ciprofloxacin (MIC 16–64 μg/mL). They had two-base substitutions in the gyrA gene at codons 83 (Ser83Phe) and 87 (Asp87Gly or Asp87Asn) and in the parC gene at codon 80 (Ser80Arg, Ser80Ile, or Ser84Lys). Our investigation documented that not only did these two S. enterica isolates have a high prevalence of ciprofloxacin resistance nationwide but also that some closely related ciprofloxacin-resistant strains are disseminated from pigs to humans.

An increasing number of clinical isolations of rapidly growing mycobacteria (RGM) at the National Taiwan University Hospital were noted from 1992 to 2001. Broth microdilution MICs of 15 antimicrobial agents were determined for 200 clinical isolates of RGM, including the Mycobacterium fortuitum group (69 isolates), M. chelonae (39 isolates), and M. abscessus (92 isolates). Our results showed that the resistance rates of these isolates to the currently available agents were remarkably high. Amikacin was active against nearly all RGM isolates. Clarithromycin was usually active against M. abscessus (79% susceptibility) and the M. fortuitum group (65% susceptibility). The majority of M. fortuitum group isolates were susceptible to ciprofloxacin (62%) and imipenem (61%). The susceptibilities to other conventional anti-RGM agents of these isolates were poor but differed markedly by species. The newer fluoroquinolones (levofloxacin, moxifloxacin, and gatifloxacin) and meropenem showed better in vitro activities against the M. fortuitum group isolates than against the other two species of RGM. Linezolid had fairly good activity against these RGM isolates, particularly against M. chelonae isolates (82% susceptible). Telithromycin had poor activity against these RGM isolates (the MICs at which 50% of the isolates tested are inhibited [MIC 50 s] were 32 to 64 μg/ml, and the MIC 90 s were >64 μg/ml).

The rapid emergence (from 0% before 1998 to 6.5% in 2000) of pandrug-resistant Acinetobacter baumannii (PDRAB) was noted in a university hospital in Taiwan. To understand the epidemiology of these isolates, we studied 203 PDRAB isolates, taken from January 1999 to April 2000: 199 from 73 hospitalized patients treated at different clinical settings in the hospital and 4 from environmental sites in an intensive-care unit. Pulsed-field gel electrophoresis analysis and random amplified polymorphic DNA (RAPD) generated by arbitrarily primed polymerase chain reaction of these 203 isolates showed 10 closely related genotypes (10 clones). One (clone 5), belonging to pulsotype E and RAPD pattern 5, predominated (64 isolates, mostly from patients in intensive care). Increasing use of carbapenems and ciprofloxacin (selective pressure) as well as clonal dissemination might have contributed to the wide spread of PDRAB in this hospital.

A total of 394 nonduplicate isolates of Streptococcus pyogenes collected from 1979 to 1998 and 267 nonduplicate isolates of Streptococcus pneumoniae collected from October, 1998, to May, 1999, in Taiwan were evaluated. Among the 220 erythromycin-resistant (MIC, > or =1 microg/ml) S. pyogenes isolates, 35% had an M phenotype and 65% had an ML phenotype (inducible resistance [iML], 0.5%, and constitutive resistance [cML], 64.5%). Among the 243 erythromycin-resistant S. pneumoniae isolates, the majority (65.4%) had an ML phenotype (iML, 0.4%, and cML, 65%) and 34.6% had an M phenotype. A substantial upsurge in the incidence of M-phenotype erythromycin-resistant isolates was found with time for S. pyogenes (0% in 1979-1984 and 100% in 1997-1998), and an increasing incidence of M-phenotype among erythromycin-resistant S. pneumoniae was also noted (

To determine the distribution and antimicrobial drug resistance in bacterial pathogens causing nosocomial infections, surveillance data on nosocomial infections documented from 1981 to 1999 at National Taiwan University Hospital were analyzed. During this period, 35,580 bacterial pathogens causing nosocomial infections were identified. Candida species increased considerably, ranking first by 1999 in the incidence of pathogens causing all nosocomial infections, followed by Staphylococcus aureus and Pseudomonas aeruginosa. Candida species also increased in importance as bloodstream infection isolates, from 1.0% in 1981-1986 to 16.2% in 1999. The most frequent isolates from urinary tract infections were Candida species (23.6%), followed by Escherichia coli (18.6%) and P. aeruginosa (11.0%). P. aeruginosa remained the most frequent isolates for respiratory tract and surgical site infections in the past 13 years. A remarkable increase in incidence was found in methicillin-resistant S. aureus (from 4.3% in 1981-1986 to 58.9% in 1993-1998), cefotaxime-resistant E. coli (from 0% in 1981-1986 to 6.1% in 1993-1998), and cefotaxime-resistant Klebsiella pneumoniae (from 4.0% in 1981-1986 to 25.8% in 1993-1998). Etiologic shifts in nosocomial infections and an upsurge of antimicrobial resistance among these pathogens, particularly those isolated from intensive care units, are impressive and alarming.

The in vitro susceptibilities of 266 isolates of Streptococcus agalactiae determined by the agar dilution method showed that 6% of isolates were nonsusceptible to penicillin and 46% was resistant to erythromycin. Of the erythromycin-resistant isolates, 86.3% had the macrolide-lincosamide-streptogramin (MLS) resistance phenotype (constitutive MLS, 85.5%; inducible MLS, 0.8%) and 13.7% had the M phenotype.

To understand quinupristin-dalfopristin resistance among clinical isolates of gram-positive bacteria in Taiwan, where this agent is not yet available for clinical use, we evaluated 1,287 nonduplicate isolates recovered from January 1996 to December 1999 for in vitro susceptibility to quinupristin-dalfopristin and other newer antimicrobial agents. All methicillin-susceptible Staphylococcus aureus (MSSA) isolates were susceptible to quinupristin-dalfopristin. High rates of nonsusceptibility to quinupristin-dalfopristin (MICs, ≥2 μg/ml) were demonstrated for the following organisms: methicillin-resistant S. aureus (MRSA) (31%), coagulase-negative staphylococci (CoNS) (16%), Streptococcus pneumoniae (8%), viridans group streptococci (51%), vancomycin-susceptible enterococci (85%), vancomycin-resistant Enterococcus faecalis (100%), vancomycin-resistant Enterococcus faecium (66%), Leuconostoc spp. (100%), Lactobacillus spp. (50%), and Pediococcus spp. (87%). All isolates of MSSA, MRSA, S. pneumoniae , and viridans group streptococci were susceptible to vancomycin and teicoplanin. The rates of nonsusceptibility to vancomycin and teicoplanin were 5 and 7%, respectively, for CoNS, ranging from 12 and 18% for S. simulans to 0 and 0% for S. cohnii and S. auricularis . Moxifloxacin and trovafloxacin had good activities against these isolates except for ciprofloxacin-resistant vancomycin-resistant enterococci and methicillin-resistant staphylococci. In Taiwan, virginiamycin has been used in animal husbandry for more than 20 years, which may contribute to the high rates of quinupristin-dalfopristin resistance.

From January 1987 through December 1998, Penicillium marneffei infection (23 patients) or colonization (1 patient) was diagnosed in a total of 24 patients in Taiwan. Of these 24 patients, 16 (67%) had AIDS and 20 (83%) had disseminated P. marneffei infection. The majority (63%) of the infections were considered indigenous. The number of cases has increased markedly in recent years, with 17 of the 24 cases diagnosed from 1996 through 1998. Twenty preserved isolates of P. marneffei, recovered from 11 patients treated at National Taiwan University Hospital during the period of January 1996 through December 1998, were studied to determine the epidemiology of P. marneffei infections. Among the 20 isolates, a total of 8 strains (highly related isolates) were identified on the basis of tests for susceptibility to 5 antifungal agents, for chromosomal DNA restriction fragment-length polymorphism types, and for randomly amplified polymorphic DNA patterns. One of the strains (6 isolates) was isolated from 4 patients treated in 1997 and 1998. Strain spreading of P. marneffei may partially contribute to the increased number of infections caused by this organism in immunosuppressed patients in Taiwan.

Objectives:To describe the epidemiology of vancomycin-resistant enterococci (VRE) in a university hospital in Taipei, Taiwan.Design:Retrospective review over a 27-month period, from March 1996 to May 1998.Setting:A tertiary-care teaching hospital in Taiwan.Participants:Patients with VRE isolated from any body site.Methods:Patients were identified through hospital microbiology and infection control records. Patient charts were reviewed for clinical and epidemiology data, including age, gender, previous hospital admissions, underlying diseases, types of infection, and recent antibiotic use. VRE isolates were characterized by their typical biochemical reactions, cellular fatty acid profiles, and the presence ofvangenes. Antibiotypes using the E-test and randomly amplified polymorphic DNA (RAPD) patterns of these isolates were used to determine the clonality.Results:Twenty-five isolates of VRE recovered from 12 patients were identified. One patient with a perianal abscess had 12 isolates of VRE (4Enterococcus faecalis, 7Enterococcus faecium, and 1Enterococcus casseliflavus) recovered from perianal lesions. Among 3 patients who were hospitalized in the same room, 1 had a community-acquired cellulitis over the left leg caused byE faecalis, and the other 2 patients both had anal colonization with 2 isolates ofE faecalis. The other 8 patients had 1E faecalisisolate each from various clinical specimens. All isolates possessed vanA resistance phenotype andvanAgenes. Different antibiotypes and RAPD patterns of the isolates from different patients excluded the possibility of nosocomial spread at the hospital.Conclusions:Multiple species of VRE (E faecalis, E faecium, andE casseliflavus) and multiple clones ofE faeciumcould colonize or infect hospitalized patients. In addition, clones of VRE can persist long-term in patients' lower gastrointestinal tracts. These results extend our knowledge of the coexistence and the persistence of multiple species and multiple clones of VRE in hospitalized patients.

From September 1990 to October 1990, 15 patients who were admitted to four different departments of the National Taiwan University Hospital, including nine patients in the emergency department, three in the hematology/oncology ward, two in the surgical intensive care unit, and one in a pediatric ward, were found to have positive blood (14 patients) or pleural effusion (1 patient) cultures for Bacillus cereus . After extensive surveillance cultures, 19 additional isolates of B. cereus were recovered from 70% ethyl alcohol that had been used as a skin disinfectant (14 isolates from different locations in the hospital) and from 95% ethyl alcohol (5 isolates from five alcohol tanks in the pharmacy department), and 10 isolates were recovered from 95% ethyl alcohol from the factory which supplied the alcohol to the hospital. In addition to these 44 isolates of B. cereus , 12 epidemiologically unrelated B. cereus isolates, one Bacillus sphaericus isolate from a blood specimen from a patient seen in May 1990, and two B. sphaericus isolates from 95% alcohol in the liquor factory were also studied for their microbiological relatedness. Among these isolates, antibiotypes were determined by using the disk diffusion method and the E test, biotypes were created with the results of the Vitek Bacillus Biochemical Card test, and random amplified polymorphic DNA (RAPD) patterns were generated by arbitrarily primed PCR. Two clones of the 15 B. cereus isolates recovered from patients were identified (clone A from 2 patients and clone B from 13 patients), and all 29 isolates of B. cereus recovered from 70 or 95% ethyl alcohol in the hospital or in the factory belonged to clone B. The antibiotype and RAPD pattern of the B. sphaericus isolate from the patient were different from those of isolates from the factory. Our data show that the pseudoepidemic was caused by a clone (clone B) of B. cereus from contaminated 70% ethyl alcohol used in the hospital, which we successfully traced to preexisting contaminated 95% ethyl alcohol from the supplier, and by another clone (clone A) without an identifiable source.

Sixty-seven clinical isolates of Streptococcus pneumoniae (40 of serotype 23F, 19 of serotype 19F, and 8 of serotype 6B) with decreased susceptibilities to penicillin and erythromycin were characterized by antimicrobial susceptibility patterns; DNA restriction endonuclease cleavage profiles of the penicillin-binding protein genes pbp1a , pbp2b , and pbp2x ; random amplified polymorphic DNA (RAPD) patterns generated by arbitrarily primed PCR; and chromosomal macrorestriction profiles based on pulsed-field gel electrophoresis. A total of 22 clones (identical or closely related pulsotypes and identical RAPD patterns) were identified; 14 clones of 23F, 6 of 19F, and 2 of 6B. Three 23F clones (26 isolates) and one 19F clone (9 isolates) expressed high-level resistance to penicillin, cefotaxime, and erythromycin (MICs ≥ 256 μg/ml). These data strongly suggest that multiple high-level penicillin-, extended-spectrum cephalosporin-, and macrolide-resistant clones of S. pneumoniae have been disseminated in Taiwan.

From 7 to 24 March 1997, four patients developed Pseudomonas fluorescens bacteremia at the hospital; one on the oncology ward and the other three in the chemotherapy room. These patients all had underlying malignancies and had the Port-A-Cath (Smiths Industries Medical Systems, Deltec, Inc., St. Paul, Minn.) implants. Three patients had primary bacteremia, and one had Port-A-Cath-related infection. None of these patients had received a blood transfusion before the episodes of bacteremia. All patients recovered: two received antimicrobial agents with in vitro activity against the isolates, and the other two did not have any antibiotic treatment. A total of eight blood isolates were recovered from these patients during the febrile episodes that occurred several minutes after the infusion of chemotherapeutic agents via the Port-A-Cath. These isolates were initially identified as P. fluorescens or Pseudomonas putida (four), Burkholderia ( Ralstonia ) pickettii (three), and a non-glucose-fermenting gram-negative bacillus (one) by routine biochemical methods and the Vitek GNI card. These isolates were later identified as P. fluorescens on the basis of the characteristic cellular fatty acid chromatogram and the results of supplemental biochemical tests. The identification of identical antibiotypes by the E test and the random amplified polymorphic DNA patterns generated by arbitrarily primed PCR of the isolates showed that the outbreak was caused by a single clone of P. fluorescens . Surveillance cultures of the possibly contaminated infusion fluids and disinfectants, which were performed 7 days after recognition of the last infected patient, failed to isolate P. fluorescens . This report of a small outbreak caused by P. fluorescens suggests that timely, accurate identification of unusual nosocomial pathogens is crucial for early initiation of an epidemiological investigation and timely control of an outbreak.

This study evaluated the clinical and microbiological characteristics of 16 patients who were colonised or infected with 26 isolates of pandrug-resistant Pseudomonas aeruginosa (PDRPA; intermediately-resistant or resistant to all cephalosporins, piperacillin–tazobactam, aztreonam, carbapenems, ciprofloxacin and aminoglycosides) in a university hospital during 1999–2002. All the isolates had colistin MICs ≤4 mg/L, 19 (73%) isolates had bla VIM-3 , and 25 (96%) isolates had class I integrons ( int I). Time-kill studies for two PDRPA blood isolates demonstrated synergism for cefepime–amikacin after 24 h. Pulsed-field gel electrophoresis analysis of the isolates revealed a polyclonal nature (12 pulsotypes), although clonal dissemination of PDRPA isolates among these patients was also present.

Escherichia coli strains are among the major bacterial causes of diarrheal illness. At least 5 categories of diarrheagenic E. coli (DEC) are recognized, namely enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), enteroinvasive E. coli (EIEC), enteroaggregative E. coli (EAEC), and enterohemorrhagic E. coli (EHEC). Due to the need for costly and labor-intensive diagnostic procedures, identification of DEC is difficult at standard laboratories. Therefore, the epidemiology of DEC infections remains obscure in Taiwan. Recently, polymerase chain reaction (PCR) or dot blot has been used for genetic detection of DEC. In this study, we analyzed 150 E. coli isolates from diarrheal stools of children under 5 years old. The PCR tests detected 5 ETEC (3.3%), 6 EPEC (4%), 4 EIEC (2.7%), and 13 EAEC (8.7%) isolates. No EHEC was detected. Dot blot and sequence analysis were used to confirm the results of PCR. The cellular fatty acid (CFA) profiles from E. coli isolates were also analyzed. Comparison of CFA composition revealed minor variation in the percentage of each fatty acid detected among DEC isolates of ETEC, EPEC, EIEC and EAEC, but did not provide enough evidence for differentiating between categories of DEC by CFA profiles alone.

Among 55 preserved isolates collected in northern Taiwan from 1999 through 2003 ciprofloxacin resistance (minimum inhibitory concentration =1 mg/mL) was found in 1 (25%) of 4 isolates obtained in 1999–2000 and in 27 (93.1%) of 29 isolates obtained in 2003. Pulsed-field gel electrophoresis analysis indicated that several clones predominated among the ciprofloxacin-resistant isolates. (excerpt)

The association of Streptococcus bovis biotypes with types of clinical infection and underlying malignancies has rarely been reported in Taiwan. The aim of this study was to characterize the clinical features and microbiological characteristics of patients with S. bovis bacteremia.From January 1992 to December 2001, 62 patients with S. bovis bacteremia were treated at National Taiwan University Hospital. Their demographic characteristics, clinical features, results of imaging studies, pathological findings, and laboratory data were retrospectively analyzed. Antimicrobial susceptibilities were determined using the agar dilution method and biotypes were determined using the API 20 Strep system.The majority of cases (76%) occurred during the 1996-1997 and 1999-2000 periods. Thirty five patients were male, and the mean age of the 62 patients was 61 years. Underlying diseases included malignancies (40%), cardiac diseases (27%), diabetes mellitus (24%), and liver cirrhosis (21%). Fifty two percent (n = 32) of patients presented with primary bacteremia and 24% (15) with definite or possible infective endocarditis. Thirteen percent (8) presented with hepatobiliary infections (acute cholecystitis and biliary tract infection). Ten patients (16%) had polymicrobial bacteremia. All of the concomitant pathogen(s) were Gram-negative rods, among which Escherichia coli predominated. The mortality rate on day 30 of illness was 21%. High Acute Physiology and Chronic Health Evaluation (APACHE) II score on the day of positive blood culture was associated with high mortality. Among the 19 patients (31%) who underwent colonoscopy, 9 (47%) had colonic lesions (tubular adenomas or carcinomas). Of the 26 patients (41%) who underwent echocardiography, 14 (54%) had vegetation in the valves. Of the 47 S. bovis isolates examined for biotypes, 37 (79%) were biotype II (29 of biotype II/2 and 8 of biotype II/1) and 10 (21%) were biotype I. The majority of isolates causing primary bacteremia (92%), hepatobiliary infections (100%) and primary bacterial peritonitis (100%) were biotype II, while 67% of isolates associated with infective endocarditis were biotype I. All isolates were susceptible to penicillin.Infective endocarditis should be highly suspected in patients with bacteremia due to S. bovis biotype I. Investigations for intra-abdominal foci other than the colon should be undertaken in patients with bacteremia caused by S. bovis biotype II. Due to the increasing number of S. bovis bacteremia patients at the hospital and unknown origins of about 50% of bacteremia cases, the need for colonoscopy and echocardiography in each case and biotyping of each blood isolate should be emphasized.

From January 1993 to December 2002, 28 patients with nutritionally variant streptococci (NVS) infections were treated at a university hospital in Taiwan. Twelve (43%) of these patients had various underlying malignancies, and 7 (25%) had underlying valvular heart diseases. Nine patients (32%) had infective endocarditis, and 9 (32%) had primary bacteremia. The deaths of 7 patients (25%) were directly related to NVS infection. Among the 28 isolates recovered from these patients, 50% were not susceptible to penicillin, 33% were not susceptible to cefotaxime, and 93% were not susceptible to azithromycin.

The incidence of nosocomial Candida fungemia increased 36-fold from 1981 (0.8/10,000 discharges) to 2000 (28.8/10,000 discharges) at the National Taiwan University Hospital, a 2000-bed teaching hospital in northern Taiwan. To understand the current status of resistance to available antifungal agents among Candida species causing invasive infections, the in vitro susceptibilities of 222 isolates (collected from July, 1999-June, 2001) were determined. Among all of the Candida species tested, 6% and 7% were resistant to fluconazole and itraconazole, respectively. The MIC90 values of voriconazole and amphotericin B were 0.5 and 1 microg/ml, respectively, although some isolates of C. krusei (amphotericin B and voriconazole MIC,64 microg/ml) and C. tropicalis and C. glabrata (voriconazole MICs,64 microg/ml) were less susceptible to voriconazole or amphotericin B. About one-half of the C. glabrata isolates belonged to susceptible dose-dependent (SDD, 36%) or resistant (12%) categories for fluconazole and 96% belonged to SDD (56%) or resistant (40%) category for itraconazole. When compared with fluconazole susceptibility data of blood Candida isolates recovered from patients treated at the same hospital (NTUH) from two different time periods (January, 1994, to June, 1995, and January, 1997, to June, 1999 described in previous reports), the incidence of increased susceptibility of non-krusei Candida isolates to fluconazole was evident. This trend of increasing susceptibility for fluconazole did not correlate to the increasing use of this agent in the hospital. None of the random amplified polymorphic DNA patterns generated by arbitrarily primed PCR using four random oligonucleotide primers for 14 isolates, which exhibited fluconazole MICs ofor = 16 microg/ml, were identical, indicating an absence of clonal dissemination among these isolates in the hospital.

Infections caused by Gram-negative bacteria with resistance to extended-spectrum cephalosporins require the identification of effective alternative antimicrobial therapy. To determine the role of other pre-existing or currently available antimicrobial agents in treating infections caused by these multidrug-resistant pathogens, we evaluated the in vitro susceptibilities of these agents in 411 non-duplicate isolates of extended-spectrum cephalosporin-resistant Gram-negative bacteria recovered between January 1999 and December 1999 in a major teaching hospital in Taipei, Taiwan. These isolates included cefotaxime-resistant (MICsor = 2 mg/L) Escherichia coli (66 isolates) and Klebsiella pneumoniae (77 isolates); cefotaxime-resistant (MICsor = 64 mg/L) Enterobacter cloacae (59 isolates), Serratia marcescens (52 isolates) and Citrobacter freundii (52 isolates); and ceftazidime-resistant (MICsor = 64 mg/L) Pseudomonas aeruginosa (50 isolates) and Acinetobacter baumannii (55 isolates). Overall, carbapenems (imipenem and meropenem) had good activity against the cefotaxime-resistant Enterobacteriaceae tested (90% of isolates were susceptible). However, carbapenems had limited activity against the ceftazidime-resistant P. aeruginosa (only 4% of isolates were susceptible) and A. baumannii (51-56% of isolates were susceptible). Among the E. coli and K. pneumoniae isolates tested, 33.3% and 58.4%, respectively, exhibited extended-spectrum beta-lactamase phenotype, determined by the double disc method. Over 80% of cefotaxime-resistant E. cloacae and C. freundii were susceptible to cefepime, but this agent had limited activity against other bacteria tested. Susceptibilities of these isolates to ciprofloxacin varied, ranging from 25% for A. baumannii to 92% for E. cloacae. Newer fluoroquinolones (moxifloxacin and trovafloxacin) had equal or less activity against these organisms, except for A. baumannii for which their MIC(90)s (8-16 mg/L) were four- to 16-fold less than that of ciprofloxacin (MIC(90) 128 mg/L).

Susceptibilities to 13 antimicrobial agents were determined by measurement of MICs for 60 isolates of Streptococcus bovis from blood cultures. Thirty-eight isolates (63.3%) had high-level resistance to erythromycin (MICs, ≥128 μg/ml). Among the 38 erythromycin-resistant strains, 21 isolates (55%) had inducible resistance to macrolides-lincosamides-streptogramin B (iMLS isolates) and 17 (45%) had constitutive resistance to macrolides-lincosamides-streptogramin B (cMLS isolates). Tetracycline resistance was also found among all of the erythromycin-resistant strains. None of the strains displayed resistance to penicillin, chloramphenicol, or vancomycin. Detection of erythromycin resistance genes by PCR and sequencing indicated that all 17 cMLS isolates were positive for the ermB gene and that 7 of 21 iMLS isolates carried the ermB gene and the remaining 14 iMLS isolates carried the ermT gene. Sequence analysis of amplified partial ermB fragments (594 bp) from S. bovis isolates revealed a 99.8% nucleotide identity and a 100% amino acid homology compared with the sequences from gene banks. The sequences of amplified fragments with primers targeted for ermC were shown to be very similar to that of ermGT ( ermT ) from Lactobacillus reuteri (98.5% nucleotide identity). This is the first report to describe the detection of the ermT class of erythromycin resistance determinants in S. bovis . The high rate of inducible erythromycin resistance among S. bovis isolates in Taiwan was not reported before. The iMLS S. bovis isolates were shown to be heterogeneous by randomly amplified polymorphic DNA analysis. These results indicate that the prevalence of inducible erythromycin resistance in S. bovis in Taiwan is very high and that most of the resistant strains carry the ermT or the ermB gene.

From January 1996 to December 1997, 200 isolates of Streptococcus pneumoniae recovered from 200 patients treated at National Taiwan University Hospital were serotyped and their susceptibilities to 16 antimicrobial agents were determined by the agar dilution method. Sixty-one percent of the isolates were nonsusceptible to penicillin, exhibiting either intermediate resistance (28%) or high-level resistance (33%). About two-fifths of the isolates displayed intermediate or high-level resistance to cefotaxime, ceftriaxone, cefepime, imipenem, and meropenem. Extremely high proportions of the isolates were resistant to erythromycin (82%), clarithromycin (90%), and trimethoprim-sulfamethoxazole (TMP-SMZ) (87%). Among the isolates nonsusceptible to penicillin, 23.8% were resistant to imipenem; more than 60% displayed resistance to cefotaxime, ceftriaxone, cefepime, and carbapenems; 96.7% were resistant to erythromycin; and 100% were resistant to TMP-SMZ. All isolates were susceptible to rifampin and vancomycin. The MICs at which 50% and 90% of the isolates were inhibited were 0.12 and 1 μg/ml, respectively, for cefpirome, and 0.12 and 0.25 μg/ml, respectively, for moxifloxacin. Six serogroups or serotypes (23F, 19F, 6B, 14, 3, and 9) accounted for 77.5% of all isolates. Overall, 92.5% of the isolates were included in the serogroups or serotypes represented in the 23-valent pneumococcal vaccine. The incidence of macrolide and TMP-SMZ resistance for S. pneumoniae isolates in Taiwan in this study is among the highest in the world published to date.

From December 1997 to March 1998, 25 methicillin-resistant Staphylococcus aureus (MRSA) isolates exhibiting negative Staphylase (Oxoid Ltd., Basingstoke, England) reactions were identified from various clinical specimens from 13 patients in six intensive care units (ICUs) or in wards following a stay in an ICU at the National Taiwan University Hospital. The characteristics of these isolates have not been previously noted in other MRSA isolates from this hospital. Colonies of all these isolates were grown on Trypticase soy agar supplemented with 5% sheep blood and were nonhemolytic and unpigmented. Seven isolates, initially reported as Staphylococcus haemolyticus (5 isolates) and Staphylococcus epidermidis (2 isolates) by the routine identification scheme and with the Vitek GPI system (bioMerieux Vitek, Inc., Hazelwood, Mo.), were subsequently identified as S. aureus by positive tube coagulase tests, standard biochemical reactions, and characteristic cellular fatty acid chromatograms. The antibiotypes obtained by the E test, coagulase types, restriction fragment length polymorphism profiles of the staphylococcal coagulase gene, and random amplified polymorphic DNA patterns generated by arbitrarily primed PCR of the isolates disclosed that two major clones disseminated in the ICUs. Clone 1 (16 isolates) was resistant to clindamycin and was susceptible to trimethoprim-sulfamethoxazole (TMP-SMZ) and was coagulase type II. Clone 2 (eight isolates) was resistant to clindamycin and TMP-SMZ and was coagulase type IV. These two epidemic clones from ICUs are unique and underline the need for caution in identifying MRSA strains with colonial morphologies not of the typical type and with negative Staphylase reactions.