Azole-resistant Candida blankii as a newly recognized cause of bloodstream infection.

Candida blankii is a newly recognized human pathogen. Here we describe a case of bloodstream infection in a preterm neonate. The yeast was repeatedly isolated from blood, and its identity was confirmed by PCR sequencing of rDNA. Additionally, C. blankii DNA was detected directly in a blood sample. The isolates initially developed pink colonies on CHROMagar Candida which later turned into dark metallic blue similar to Candida tropicalis. Inaccurate identification by the VITEK 2 yeast identification system as Stephanoascus ciferrii and intrinsic resistance to fluconazole (MIC 12-16 µg/mL) underscore the need for its accurate identification for appropriate therapeutic management.

Persistent Candida conglobata bloodstream infection in a preterm neonate successfully treated by combination therapy with amphotericin B and caspofungin.

Fungemia due to uncommon/rare Candida species is an emerging problem of global clinical significance. Here, we describe a case of Candida conglobata bloodstream infection in a preterm neonate. The diagnosis was established by repeated isolation of C. conglobata in blood cultures and by detection of rDNA of the fungus in serum samples. The identity of the isolate as C. conglobata was confirmed by sequencing of ITS region and D1/D2 domains of rDNA. Despite initial treatment with a liposomal amphotericin B (AmBisome) for 7 days, the blood culture remained positive. The neonate was successfully treated by combination therapy with caspofungin for 25 days. To the best of our knowledge, this is the first proven report unequivocally proving the etiologic role of C. conglobata in bloodstream infection.

High prevalence of toxic shock syndrome toxin-producing epidemic methicillin-resistant Staphylococcus aureus 15 (EMRSA-15) strains in Kuwait hospitals.

This study characterized EMRSA-15 isolates obtained from patients in Kuwait hospitals for their genotypic relatedness, antibiotic resistance and carriage of virulence genes using pulsed-field gel electrophoresis (PFGE), coagulase serotyping, SCCmec subtyping, spa typing, multilocus sequence typing and DNA microarray. The isolates were resistant to trimethoprim (75.6%), ciprofloxacin (29.7%), erythromycin and clindamycin (24.3%), tetracycline (19.0%), and gentamicin and kanamycin (21.6%). All 37 isolates belonged to sequence type (ST) 22, coagulase type XI, three PFGE types and eight subtypes, ten spa types including t223 (51.3%), t852 (13.5%), t032 (8.1%), t790 (8.1%), t3107 (5.4%) and one each of t309, t2251, t3935, t5708 and t5983. Twenty-six isolates (70.2%) carried SCCmec IVa, eight isolates carried SCCmec IV and three isolates carried SCCmec IVh. All isolates carried agr1, cap5 and egc gene cluster (seg, sei, selm, seln, selo, and selu). tst (toxic shock syndrome toxin) was detected in 23 isolates. Eight isolates (21.6%) were positive for Panton-Valentine leukocidin (PVL). Genotypic analysis revealed that 62.1% of the isolates comprising ST22-IVa-t223 (51.3%) and ST22-IVa-t309/t2251/t3935/t5708 (10.8%) were CC22-[tst1(+)] UK EMRSA-15/Middle Eastern variant, 21.6% were CC22-PVL(+) EMRSA-15 variant and 16.2% were CC22-UK EMRSA-15/Barnim clone. These results show that the tst1 positive-ST22-IVa-t223 (Middle Eastern variant) and the CC22-PVL(+) EMRSA-15 variant were the dominant EMRSA-15 variants in Kuwait hospitals.

First isolation of Candida metapsilosis in Kuwait, an emerging global opportunistic pathogen.

Invasive infections due to uncommon and rare yeast species are increasing worldwide in prevalence and are associated with high mortality rates. Here, we describe the first isolation and characterization of Candida metapsilosis cultured from the blood sample of a 10-year-old Saudi girl, who suffered from a neurodegenerative disorder, in Kuwait. The yeast isolate was identified by sequencing of ITS region and D1/D2 domains of rDNA. The report extends the geographic distribution of C. metapsilosis to the Middle East and highlights the emerging role of uncommon yeast species causing infections in susceptible hosts.

Three distinct clones of carbapenem-resistant Acinetobacter baumannii with high diversity of carbapenemases isolated from patients in two hospitals in Kuwait.

OBJECTIVES: This study was undertaken to investigate the clonal relatedness of multidrug-resistant (MDR) Acinetobacter baumannii isolates collected from patients in two teaching hospitals in Kuwait. MATERIALS AND METHODS: Clinically significant consecutive isolates of A. baumannii obtained from patients in the Mubarak (36) and Adan (58) hospitals over a period of 6 months were studied. These isolates were identified using molecular methods, and their antimicrobial susceptibility was determined by the Etest method. The mechanism of resistance to carbapenem was investigated by PCR, and pulsed-field gel electrophoresis (PFGE) was used to determine the clonal relatedness of MDR isolates. RESULTS: Of the 94 isolates investigated, 80 (85.1%) were multidrug resistant (MDR). The A. baumannii PFGE clone A and subclone A1 were the most prevalent in patients infected with MDR isolates. Fifty-five (94.8%) and 15 (41.7%) of the MDR isolates from the Adan and Mubarak hospitals, respectively, belonged to PFGE clone A; isolates in this group showed higher resistance rates to antibiotics than isolates form other groups. Of the 94 isolates, 40 (42.6%) were resistant to either imipenem or meropenem or to both (CRAB). Most CRAB isolates (29/40 or 72.5%) carried bla genes, which code for MBL (VIM-2 and IMP-1) enzymes. Two isolates harbored bla(OXA-23). CONCLUSION: Three distinct clones of CRAB were isolated, providing evidence of a high diversity of carbapenemases among our geographically related isolates.

The prevalence of Chlamydia trachomatis and Neisseria gonorrhoeae infections among men with urethritis in Kuwait.

PURPOSE: Chlamydial non-gonococcal urethritis and gonorrhoea are the most common sexually transmitted bacterial infections worldwide. Data on these infections are scanty in the Islamic world, especially Kuwait. The objective of this study was to establish the prevalence of Chlamydia trachomatis and Neisseria gonorrhoeae among men with symptomatic urethritis in Kuwait. METHODS: Men with urethral discharge seen and managed at eight governmental hospitals were recruited into the study. A pair of urethral swab and first-voided urine sample were taken from the patients and sent immediately to the laboratory where they were processed using strand displacement nucleic acid amplification kits (SDA; ProbeTec, Becton Dickinson); one pair per patient was studied. RESULTS: A total of 426 symptomatic men were studied, out of whom 155 (36.4%) were infected by either C. trachomatis or N. gonorrhoeae, or both. The overall prevalence rates of C. trachomatis and N. gonorrhoeae were 12.4% and 23.9%, respectively. There was no significant difference in chlamydial and gonococcal prevalence between Kuwaiti men and non-Kuwaitis (P>0.05). Infection rates were much lower in married men than unmarried men. Men in the age range of 21-35 years were more vulnerable to both infections. CONCLUSION: The findings show that N. gonorrhoeae and, to a lesser extent, C. trachomatis are common in men with urethritis in Kuwait. Appropriate preventive strategies that conform to Islamic rules and values should be of highest priority of the policymakers.

An outbreak of CTX-M-15-producing Klebsiella pneumoniae isolates in an intensive care unit of a teaching hospital in Kuwait.

OBJECTIVE: This study reports an outbreak of Klebsiella pneumoniae infections in 14 patients during a 2-month period (August-September, 2008) in the intensive care unit (ICU) of a teaching hospital in Kuwait. MATERIALS AND METHODS: The clinical sources were blood (9), urine (3) and respiratory secretions (2) identified by the automated VITEK-2 ID System. Susceptibility testing was performed by the E-test method. Extended-spectrum ß-lactamase (ESBL) production was assessed using the ESBL E-test and confirmed by PCR. Carriage of bla genes was determined by PCR and sequence analysis. The transferability of resistance phenotypes was demonstrated by conjugation experiments and clonal relatedness was determined by PFGE. RESULTS: The isolates were susceptible to imipenem, meropenem, and tigecycline and produced ESBL. All isolates yielded an amplicon of 499 bp with universal consensus primers (MA primers). DNA sequence analysis showed that they all harboured bla CTX-M-15 and bla TEM-1 genes. The environmental isolate obtained from a suction machine was also CTX-M-15/TEM-1 producer. The resistance phenotypes were transferrable to the Escherichia coli J53 r strain. PFGE, revealed two clones, A and B, related with a Dice coefficient of >94.1%. A mortality rate of 21.4% was recorded. CONCLUSION: The outbreak was contained by robust and aggressive infection control measures. This study highlights the first outbreak of CTX-M-15-producing K. pneumoniae associated with high mortality in an adult medical ICU in Kuwait.

Emergence of tigecycline and colistin resistance in Acinetobacter species isolated from patients in Kuwait hospitals.

The development of resistance is a compelling reason for reviewing administration of antibiotics. Recently, most Acinetobacter infections are caused by multidrug-resistant (MDR) strains which have necessitated the use of tigecycline or colistin. This study was undertaken to determine the susceptibility of Acinetobacter spp. to these and other drugs. A total of 250 Acinetobacter isolates were collected from the 8 government hospitals over a period of 6 months. Susceptibility to 18 antibiotics, including tigecycline and colistin, was investigated by determining their minimum inhibitory concentrations using E test. Of the 250 isolates, 13.6% and 12% were resistant to tigecycline and colistin. A total of 25.2% and 37.2% were resistant to imipenem and meropenem, respectively. Of the 250 isolates 88.4% were MDR. This relatively high prevalence of tigecycline and colistin-resistant isolates indicates an emerging therapeutic problem which may severely compromise the treatment of MDR Acinetobacter spp. infections in Kuwait.

Sequence analysis of bla(CTX-M) genes carried by clinically significant Escherichia coli isolates in Kuwait hospitals.

OBJECTIVE: To investigate the extent, distribution and sequence analysis of bla(CTX-M) genes carried by Escherichia coli isolated from patients admitted to all government hospitals in Kuwait. METHODS: Extended-spectrum ß-lactamase (ESBL)-producing E. coli isolates were collected from the 8 major hospitals in Kuwait. CTX-M ESBLs were analyzed by PCR and sequenced. Clonality of the positive isolates was determined for genetic relatedness using pulsed-field gel electrophoresis (PFGE) with XbaI digestion of the genomic DNA. RESULTS: Of the 136 ESBL-positive isolates, 106 (77.9%) harbored bla(CTX-M) genes. Among these, bla(CTX-M-15) was the most frequent with a prevalence rate of 84.1%, followed by bla(CTX-M-14) (6.8%), bla(CTX-M-14b) (5.7%) and bla(TOHO-1) (3.4%). Ninety-three (87.7%) were isolated from Kuwaiti (35.9%), Egyptian (31.1%) and Indian (20.8%) nationals; the majority of isolates positive for bla(CTX-M-15) were mainly from these 3 nationalities. PFGE analysis did not demonstrate any clustering of positive isolates in any particular hospital. CONCLUSION: This study confirms an explosive emergence of CTX-M-15 ß-lactamase among E. coli isolates in Kuwait and shows that the strains were clonally heterogeneous with no evidence of inter- or intra-hospital spread. Thus Kuwait may represent an important source of CTX-M-15-positive E. coli.

National surveillance of antimicrobial susceptibility of CTX-M-positive and -negative clinical isolates of Escherichia coli from Kuwait government hospitals.

Antibiotic resistance in Escherichia coli is becoming a complex therapeutic problem. Surveillance programs are valuable tools and offer important information on bacterial resistance trends. This study was undertaken to determine the susceptibility of clinically significant isolates of E. coli obtained from patients admitted to 8 Kuwait government hospitals and to examine how this was influenced by the production of CTX-M extended-spectrum beta-lactamases (ESBLs). The susceptibility of 876 consecutive clinically significant strains of E. coli to 13 antibiotics was determined by Etest. ESBL production was assessed by ESBL-Etest method and the presence of CTX-M beta-lactamases was confirmed by PCR technique. Of the 876 isolates, 604 (69%) were highly non-susceptible to ampicillin with MIC(90 )of >256 microg/ml. Resistance to the 3(rd)-generation cephalosporins ranged from 7.5% in the Maternity hospital to 29% in the Ibn Sina hospital; ciprofloxacin resistance rates ranged from 14% and 40%, respectively. Carbapenems and amikacin demonstrated excellent activity. The minimum inhibitory concentrations (MIC(90)) of cefotaxime, ceftazidime, cefepime and ciprofloxacin were >256, 64, >256 and >32 microg/ml, respectively for CTX-M-positive isolates versus 0.5, 1, 025 and 0.125 microg/ml for CTX-M-negative strains. Frequencies of CTX-M-positive isolates within the cefotaxime MIC ranges of 1-2, 3-8, 9-16 and >16 microg/ml were 0, 4, 15 and 81%, respectively. In conclusion, the susceptibility of E. coli to the 3(rd )generation cephalosporins and ciprofloxacin was influenced by the presence of CTX-M ESBL and a high proportion of the CTX-M-producing isolates were in the susceptibility ranges of cefotaxime.

Emergence of resistance to amphotericin B and triazoles in Candida glabrata vaginal isolates in a case of recurrent vaginitis.

Emergence of resistance to triazoles and amphotericin B in Candida glabrata vaginal isolates is documented by Etest. During the 18-month follow-up of a case of vaginitis, 14 consecutive isolates of C. glabrata were examined. The isolates exhibited development of in vitro resistance beginning with itraconazole (>32 microg/ml), followed by fluconazole (>256 microg/ml), amphotericin B (>32 microg/ml), and voriconazole (>32 microg/ml). The DNA sequence analyses and finger printing of the isolates strongly suggest that our patient remained colonized with a single strain. The report underscores the propensity of C. glabrata to acquire resistance during antifungal therapy and the importance of susceptibility testing in the management of infections caused by this species.

Genetic lineages of community-associated methicillin-resistant Staphylococcus aureus in Kuwait hospitals.

Twenty-six community-associated methicillin-resistant Staphylococcus aureus (CAMSRA) isolates were characterized by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) and screened for accessory gene regulator (agr), capsular polysaccharide (cap), and Panton-Valentine leucocidin (PVL) genes. They exhibited five PFGE patterns (types A to E). The majority were PFGE type A (12 isolates) or type B (8 isolates). MLST showed that PFGE type A isolates belonged to sequence type 80 (ST80), while the PFGE type B isolates were ST30. The ST80 and ST30 clones contained agr allotype 3, cap type 8, and PVL. The results showed that two internationally recognized CAMRSA clones are dominant in Kuwait hospitals.

Surveillance of antibacterial resistance in Staphylococcus aureus isolated in Kuwaiti hospitals.

OBJECTIVE: To investigate the prevalence of antibiotic resistance among Staphylococcus aureus isolated in Kuwaiti hospitals. MATERIALS AND METHODS: S. aureus were isolated and identified following standard microbiological methods. Antibacterial susceptibility test was performed by disk diffusion and the measurement of minimum inhibitory concentration with E-test strips. RESULTS: A total of 1,846 S. aureus isolates were analyzed from 13 hospitals between 1 March and 30 October 2005. They were isolated from 1,765 (95.6%) inpatients and 81 (4.4%) outpatients. Methicillin resistance was detected in 588 (32.0%) of the isolates. The methicillin-resistant S. aureus (MRSA) consisted of 461 (78%) multiresistant and 127 (22%) nonmultiresistant isolates. The nonmultiresistant MRSA consisted of epidemic MRSA-15 and community-associated MRSA. The community-associated MRSA was detected in all hospitals with MRSA, indicating its establishment in Kuwaiti hospitals. The proportion of isolates resistant to gentamicin, kanamycin, erythromycin, tetracycline, ciprofloxacin, fusidic acid and trimethoprim was higher among MRSA than methicillin-susceptible S. aureus (MSSA) isolates. Twenty-four and 22% of MRSA and MSSA isolates, respectively, expressed reduced susceptibility to vancomycin (minimum inhibitory concentration = 3-4 mg/l). CONCLUSION: The study revealed the presence of methicillin resistance in 32% of S. aureus isolated in Kuwaiti hospitals and revealed an increase in the number of MRSA and MSSA with reduced susceptibility to vancomycin.

Characterisation of non-multiresistant methicillin-resistant Staphylococcus aureus (including EMRSA-15) in Kuwait Hospitals.

This study characterised non-multiresistant methicillin-resistant Staphylococcus aureus (nmMRSA) isolates from Kuwait hospitals to ascertain whether they were community-acquired MRSA (CA-MRSA). Forty-two nmMRSA isolates obtained between July 2001 and October 2003 were analysed by staphylococcal cassette chromosome mec (SCCmec) typing, bacteriophage typing, production of Panton-Valentine leukocidin (PVL), urease and staphylococcal enterotoxins A, B, C and D, TSST-1, and by pulsed-field gel electrophoresis (PFGE). Forty-one isolates were SCCmec type IV, and one isolate was SCCmec type III. The isolates belonged to six PFGE patterns, with two types, A and D, distributed in six and four hospitals, respectively. Most (n = 26; 61.9%) isolates produced urease. These isolates were mainly from wound and skin infections, showed low-level methicillin resistance (MIC 8-48 mg/L), and nine carried genes for PVL. These characteristics, together with their carriage of the type-IV SCCmec, identified the isolates as CA-MRSA. Ten of the 16 urease-negative isolates produced staphylococal enterotoxin C; 12 reacted weakly with phage 75, and were resistant to clindamycin and/or erythromycin, which are characteristics of EMRSA-15. Thus, this study identified the co-existence of two types of nmMRSA, i.e., CA-MRSA and EMRSA-15, in Kuwait hospitals.

Antibacterial resistance and molecular typing of methicillin-resistant Staphylococcus aureus in a Kuwaiti general hospital.

OBJECTIVE: To investigate antibiotic resistance and genetic relatedness of methicillin-resistant Staphylococcus aureus (MRSA) isolated in a general hospital in Kuwait over a period from 1996 to 1998 and 2001. MATERIAL AND METHODS: The isolates were characterized by antibacterial susceptibility testing, coagulase serotyping, coagulase gene polymorphism (coag-RFLP) and pulsed-field gel electrophoresis (PFGE). RESULTS: The MRSA isolates were highly resistant to gentamicin, kanamycin, ciprofloxacin, tetracycline, fusidic acid and mupirocin. The prevalence of gentamicin, kanamycin, streptomycin, tetracycline and erythromycin resistance remained high (80-96%) throughout the study period, but the prevalence of resistance to ciprofloxacin, fusidic acid and mupirocin steadily increased. The already high mupirocin resistance level increased from 12.5 in 1996, to 85.7% in 2001, and the fusidic acid resistance varied between 70.8 and 85.7%. In contrast, chloramphenicol and trimethoprim resistance declined from 25 and 29% in 1996 to 4.7 and 14.2% in 2001, respectively. The majority (91.5%) of the isolates were coagulase serotype 4. AluI restriction endonuclease analysis of amplified coagulase gene generated four coag-RFLP patterns: 92% of them were coag-RFLP type 1, while types 2, 3 and 4 were 3.5, 4.6 and 1.1% respectively. PFGE differentiated them into seven pulsotypes (PFGE types 1-7). The PFGE type 1 pulsotype constituted 90.2% of the isolates. Isolates with the type A coag-RFLP also had the type1 PFGE pulsotypes. CONCLUSION: The concordant results of PFGE and coag-RFLP demonstrated the presence of a persistent MRSA clone in the hospital during the study period.

Sunflower seed husk agar: a new medium for the differentiation of Candida dubliniensis from Candida albicans.

A sunflower (Helianthus annuus) seed husk agar medium has been developed and evaluated for differentiation of Candida dubliniensis from Candida albicans on the basis of colony morphology and chlamydospore production. All C. dubliniensis isolates (n=40) produced rough colonies with hyphal fringes and abundant chlamydospores whereas 101 of 105 (96.2%) C. albicans isolates produced smooth colonies with no evidence of chlamydospore production. Since this medium is free from oil droplets, chlamydospores can be examined with greater clarity by Dalmau plate technique. This medium provides a simple and cost-effective tool for the presumptive differentiation of C. dubliniensis from C. albicans and is particularly suited for clinical microbiology laboratories where biochemical or molecular methods for the differentiation of these two species are not available.

Antibiotic susceptibility profile of group B streptococcus (Streptococcus agalactiae) at the Maternity Hospital, Kuwait.

OBJECTIVES: This study was designed to determine the antibiotic susceptibility profile of clinical isolates of group B streptococcus(GBS, Streptococcus agalactiae) and to use the information for formulating appropriate intrapartum antibiotic policy for GBS carriage in pregnancy. MATERIALS AND METHODS: A total of 1,166 clinical isolates (single isolates) of GBS obtained from vaginal/rectal samples of pregnant mothers seen at the Maternity Hospital, Kuwait were studied over a period of 43 months between May 1998 and December 2001. The isolates were identified by standard methods and their susceptibility to penicillin, ampicillin, cephalothin, clindamycin and erythromycin was determined by disk diffusion technique, minimal inhibitory concentration (MIC) using the Vitek automated sensitivity card system and E-test methods. RESULTS: All the GBS isolates were fully susceptible to penicillin, ampicillin and cephalothin. Only 0.7 and 1.7% were resistant to erythromycin and clindamycin, respectively. Disk diffusion results interpreted by the standard interpretative criteria recommended by the National Committee on Clinical Laboratory Standards correlated well with Vitek results as well as the E-test for penicillin. The MIC of penicillin against all isolates ranged between 0.016 and 0.064 microg/ml. For the 6 months of 1998 and throughout 1999, the percentages of isolates susceptible at MICs of 0.016, 0.023, 0.032, 0.047 and 0.064 microg/ml were 6.5, 9.9, 31, 38.8 and 12%, respectively. The trend was similar in the subsequent years except that the percentage of isolates susceptible at MIC of 0.064 microg/ml increased to 26.6% in 2000, but went down to 4.4% in 2001. CONCLUSION: The trend in susceptibility of GBS to a variety of often used antibiotics for therapy and prophylaxis remained unchanged over nearly a 4-year period. The apparent increase in the number of isolates susceptible at higher MIC values of penicillin (0.047 and 0.064 microg/ml) in 2000 appears to be a bleb that cannot be explained by any event in the hospital for that year. Our data, based on susceptibility profiles, supports the use of penicillin or ampicillin for intrapartum chemoprophylaxis to prevent early-onset neonatal GBS infections.

In vitro activity of linezolid and other antibiotics against Gram-positive bacteria from the major teaching hospitals in Kuwait.

Multidrug-resistant Gram-positive bacteria are an increasingly pressing problem in the clinic. Consequently, linezolid, a recently introduced oxazolidinone with Gram-positive activity, was tested in comparison with 10 other antibiotics against 8103 clinically significant Gram-positive cocci by Etest, disk diffusion and Vitek methods. Linezolid demonstrated excellent activities against all isolates. Vancomycin and teicoplanin demonstrated equally excellent activity against almost all isolates. The methicillin-resistant Staphylococcus aureus (MRSA) strains were all susceptible to the glycopeptides and linezolid, but resistant to erythromycin (96%), fusidic acid (91.5%), gentamicin (84%) and clindamycin (73%). Forty one and 39% of the Streptococcus pneumoniae isolates were resistant to penicillin (MIC >0.5 microg/ml) and erythromycin (MIC >1 microg/ml), respectively. S. agalactiae susceptibility was 9% and 10% resistant to clindamycin and erythromycin, respectively. In conclusion, all the Gram-positive isolates tested were susceptible to linezolid. With its oral bioavailability profiles, it obviously holds great promise. Our data should be a useful addition to the literature from the Middle East.

Characterization of high-level aminoglycoside-resistant enterococci in Kuwait hospitals.

This study investigated the distribution of genes for aminoglycoside-modifying enzymes (AME) and the genetic relatedness of high-level aminoglycoside-resistant enterococci isolated in Kuwait hospitals. A total of 117 enterococci, consisting of 109 Enterococcus faecalis, seven Enterococcus faecium, and one Enterococcus casseliflavus were studied. The MICs of gentamicin, kanamycin, amikacin, tobramycin, and streptomycin were determined by agar dilution and the genes encoding the AAC(6')- APH(2"), ANT(4'), APH(3'), APH (2")-Ib, APH (2")-Ic, APH (2")-Id, and ANT(6) enzymes were amplified by PCR. They were typed by pulsed-field gel electrophoresis (PFGE). Filter mating was used to transfer gentamicin resistance determinants. They were all resistant to kanamycin (MIC 2000 mg/L). Fifty-five isolates were resistant to gentamicin (MIC 500 mg/L), 72 were resistant to tobramycin (MIC 64 mg/L), 115 were resistant to amikacin (MIC 64 mg/L), and 97 were resistant to streptomycin (MIC 1000 mg/L). The aac(6')-Ie-aph(2")-Ia was detected in all isolates with gentamicin MIC 500 mg/L and in 15 isolates with gentamicin MIC 256 mg/L. The aph(3')-IIIa gene was detected in 101 isolates, whereas the ant(6')-Ia gene was detected in 85 of the 97 streptomycin-resistant isolates with MIC 1000 mg/L. The aac(6')-Ii gene was detected only in the seven E. faecium isolates. None of them contained ant(4')-Ia, aph(2")-Ib, aph(2")-Ic and aph(2")-Id. PFGE revealed heterogeneous patterns with no dominant clone. The results demonstrated that AME are common in aminoglycoside-resistant enterococci isolated in Kuwait. However, the absence of a dominant clone suggests that they acquired high-level aminoglycoside independently.

A chromosomal location of the mupA gene in Staphylococcus aureus expressing high-level mupirocin resistance.

OBJECTIVES: To investigate the genetic location of the mupA gene in high-level mupirocin-resistant Staphylococcus aureus isolates. MATERIALS AND METHODS: Antibiotic resistance was detected by disc diffusion. The Etest was used to determine mupirocin MIC. The presence of mupA was detected by PCR using specific primers. Curing, transfer experiments, pulsed-field gel electrophoresis (PFGE) and DNA hybridization were used to study the genetic location of mupA. RESULTS: The isolates had mupirocin MICs > 1024 mg/L and were resistant to methicillin, gentamicin, kanamycin, streptomycin, erythromycin, tetracycline, ciprofloxacin, cadmium acetate, propamidine isethionate and ethidium bromide. They carried two plasmids of approximately 26 and 2.8 kb. Curing and transfer experiments demonstrated that the 26 kb plasmid encoded resistance to cadmium acetate, propamidine isethionate and ethidium bromide. Loss of mupirocin resistance corresponded to the loss of a 40 kb DNA fragment from a 175 kb SmaI chromosomal fragment. The mupA gene was detected only in the genomic DNA of the mupirocin-resistant strains and in their derivatives cured of the 26 kb plasmid. A labelled mupA probe hybridized to the 175 kb SmaI fragment only in the mupirocin-resistant isolates. CONCLUSION: The absence of mupA on any of the plasmids and its detection only in the chromosomal DNA of the parents and in their derivatives cured of the 26 kb plasmid strongly supports a chromosomal location for mupA in these isolates.