While we confirmed only one vre strain among our isolates, noteworthy in dubai reviewing the literature was that the Aga Khan University hospital clinical lab in Karachi reported seven vre among inpatients in 1997, none in outpatients, 12 and again none among both inpatients and outpatients. The intrinsic ruggedness of enterococci also confers an unusual ability to acquire resistance to such diverse groups of drugs as aminoglycosides, beta-lactams, macrolides, fluoroquinolones and tetracyclines. 2,8 Accordingly, ampicillin, ciprofloxacin and erythromycin, along with cotrimoxazole, doxycycline and meropenem performed poorly, as did gentamicin. Though not indicated, we included erythromycin and doxycycline profiles as a note in particular for dermatologists who empirically prescribe these drugs. Nitrofurantoin affected all but 3 of the 112 isolates, suggesting that it could be a drug of choice for treating enterococcal uti. Indeed, nitrofurantoin levels in urine are said to reach concentrations of 200-400 ug/mL, which are well above the mics of susceptible uropathogens, and in one report was said to be active on all 300 enterococcal isolates from a hospital, a third of which were resistant. 13 While nitrofurantoin is primarily a urinary antiseptic, chloramphenicol can be employed for treating diverse infected sites in the body; it influenced as many.5 of our strains in vitro including the vre isolate, and may well be noted as a possible choice for. Others have suggested daptomycin, 14 dalbavancin, 15 and ramoplanin 16 especially for vre strains, but these drugs are hardly available in pakistan, and, being expensive, often beyond reach of the unaffording patient.
It was also indifferent to teicoplanin (mic 32ug/ml a drug which was effective on all the other strains (MICs 2ug/mL). However, it was sensitive to chloramphenicol and nitrofurantoin in Etest mic evaluation, and in addition also to the standard fosfomycin disc which gave an excellent zone of inhibition ( 20mm) in the disc-diffusion assessment. Also, the vre isolate was unaffected by routine discs of doxycycline, meropenem and pipericillin-tazobactam which afforded poor inhibitory zones well below the required resistance cut-off diameters of 12mm, 13mm and 17mm respectively. Discussion illness caused by the genus Enterococcus (notably. Faecalis, which accounts for almost writing 80 of all infections) 1 include, among others, uti and bacteremia. In our study, the isolate was grown mainly from urine (69.6 followed by pus and blood. All the 112 isolates were challenged with 13 representative drugs irrespective of the site of infection to ascertain their general susceptibility profiles in our environment where "over-the-counter" drug vending and empirical prescribing abound. Vancomycin insusceptibility is an acquired resistance mediated by transposons or plasmids in bacteria, and these can develop in hospital wards that regularly use the drug and initiate serious infections. The single vre encountered in our study was grown from the urine of an elderly male following cardiac bypass surgery and catheterization, suggesting that the mechanism of genetic exchange was likely at play in his sickbay.
Results comparison of disc diffusion zone diameters: Results (Table-I) indicate that vancomycin and teicoplanin were significantly effective on all but one of the 112. The single vre strain, encountered in the urine of an elderly hospitalized male, was unaffected by the 30ug vancomycin disc and was also indifferent to teicoplanin. All the other isolates were sensitive to vancomycin, and "Intermediate" vancomycin resistance was also not seen. Nitrofurantoin and fosfomycin inhibited 109 and 102 of the isolates respectively, followed in effectiveness by chloramphenicol and pipericillin-tazobactam. Ampicillin, mero- penem and ciprofloxacin offered unsatisfactory inhibition zones. Dalbamycin, daptomycin, linezolid, pristinamycin, ramoplanin and quinopristin/dalfopristin discs and Etest Epsilometers were not available in the local market for evaluation. Mics of the vre isolate and related sensitivity profiles: Table-ii illustrates the Etest mic sensitivity profile of the single vre strain encountered. It showed "low level" resistance to vancomycin (08ug/mL) and was resistant to ampicillin, ciprofloxacin, erythromycin, gentamicin and cotrimoxazole.
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The isolates from urine (n78 pus (n18 blood (n12 hvs (n2) and ascitic fluid (n2) also grew in broth containing.5 nacl incubated at 35C for 3 days, fermented aesculin on Nutrient agar with 4 bile (Oxoid, uk and were arabinose negative, pyruvate and sorbitol. The patients (n112) included adults of both sexes aged 19-72, a male neonate and a 6 year-old boy. The following available antimicrobial agents (n9) were obtained as pdm epsilometers (ab biodisk, solna, sweden) for Etest mic estimation: ampicillin, chloramphenicol, ciprofloxacin, cotrimoxazole, erythromycin, gentamicin, nitrofurantoin, teicoplanin, and vancomycin. Isolated colonies of the single vre isolate from a 24hr plate were homogenized in suspension medium to achieve.5 McFarland turbidity and spread on mueller Hinton agar (Oxoid, uk). Etest strips were promptly positioned using a 6-strip template, the plates incubated at 35C for 24hrs, and the inhibition marketing intersection in the ellipses recorded.
Isolates with mic breakpoints of 16ug/mL for ampicillin and 08ug/mL for vancomycin were considered to be resistant; nitrofurantoin susceptibility was taken as 32 ug/mL. Faecalis atcc 29212 was used as the control strain. Sensitivity screening with commercial discs (Oxoid, uk) included the above nine antibiotics along with doxycycline (30ug fosfomycin (50ug meropenem (10ug) and pipericillintazobactam (110ug) were performed on mueller-Hinton agar (Oxoid, uk) using the standard disc diffusion technique. 10 However, for gentamicin, along with the usual 10ug disc, an additional 120ug disc was added to check for high-level aminoglycoside resistance, with the resultant zone diameter 06mm considered resistant, and 10mm sensitive. 11 Results of ampicillin tests (disc content: 10ug, with zones 16mm resistant and 17mm sensitive) also predicted the susceptibility to amoxicillin and pipericillin. For testing against vancomycin, a disc of 30mg was used and the zones of inhibition measured after an incubation time of 24hrs using transmitted light. 11 Presence of any growth within the zone indicated possible resistance, and mics of the implicated isolate were determined.
Introduction, enterococci comprise a significant portion of the normal intestinal flora, with some also being found on the skin, in oropharyngeal and vaginal secretions and in the perineal area. 1, nevertheless, they have become increasingly important nosocomial pathogens in recent years as a result of increased incidence of resistance to many antimicrobials. 2, enterococcal infections most commonly occur in the urinary tract, but the organism also crops up in bacteremia, wound infections, intra-abdominal abscesses, infective endocarditis and infrequently, meningitis. 3, the intrinsic or relative resistance of the isolates to aminoglycosides, cephalosporins and cotrimoxazole, and the ability to acquire resistance to erythromycin and tetracycline is well known. 4, ampicillin was the drug of choice for enterococcal uti, but resistance to this agent has been rapidly emerging. Also, the fluoroquinolones are said to be marginally effective on enterococci, clindamycin is generally considered to be inactive at clinically achievable levels, and the action of cotrimoxazole is compromised in vivo due to endogenous sources of folate.
2, the recent disturbing trait to emerge is indifference to vancomycin 5 necessitating the search for alternative drugs 6 and the exploitation of various synergistic combinations of selected antibiotics. While deep tissue and bloodstream infections have been traditionally treated with ampicillin-gentamicin, or with vancomycin-gentamicin, high level aminoglycoside resistance has encouraged alternative therapy. A beta-lactam with daptomycin has been employed, 7 as has quionopristin/dalfopristin. 8 But a review of 69 cases of vre found that none of the 42 different combinations of antimicrobials used were absolutely effective, 9 which illustrates the extent of the problem in handling relevant clinical infections. The final prescription, then, is prejudiced by the drug-sensitivity profile of local isolates, as well as the site of the enterococcal infection involved. Hence, our study screening 112 consecutive clinical isolates was designed to fathom the extent of multidrug-resistance of enterococci in our environment to easily available antibiotics, and thereby suggest possible options and limitations of drug prescription. Material and methods a total of 103 isolates identified as Enterococcus faecalis from specimens submitted for culture by city hospitals, and 09 referred by private physicians, were processed on sheep-blood agar (Merck, hamburg). Essas Lab which has its branches in key areas of Karachi metropolis, during a 2-year period ending September, 2005.
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Conclusion: Increasing drug resistance of enterococci warrants concern and the search for possible therapeutic options prejudiced by local patterns of resistance and the site of infection. Vancomycin, teicoplanin, nitrofurantoin and chloramphenicol are also significantly effective on our current isolates and fosfomycin, typically considered a urinary drug, but help encouraged by reports of its successful use in diverse sites, is discussed as a possible alternative. Key words: Enterococci, vre, treatment options, nitrofurantoin, fosfomycin. Pak j med Sci july - september 2006 Vol. Essas Laboratory and diagnostic Center, karachi - pakistan. C-87, Block-15 Gulistan-e-jouhar, karachi, pakistan. Email: received for Publication: December 28, 2005 revision Received: March 30, wallpaper 2006 revision Accepted: April 10, 2006.
enterococci to many antimicrobials poses a major therapeutic challenge; vancomycin-resistant enterococci (vre in particular, exhibit variation in resistance patterns, demanding tailored drug prescription. Hence we screened the in vitro sensitivity profiles of 112 local clinical isolates using 13 antibiotics for their possible therapy options biased by the site of infection. Setting: Specimens yielding enterococci referred by local hospitals (n103) and private physicians (n09) were processed during a two-year period ending September 2005. Essas Lab in Karachi, pakistan. Methods: Consecutive, enterococcus faecalis strains cultured from urine (n78 pus (n18 blood (n12 hvs (n2) and ascitic fluid (n2) were challenged in vitro with locally available antibiotics using the standard disc diffusion method; mics of one vre isolate were also performed. Results: Vancomycin (99.1) and teicoplanin (99.1 followed by nitrofurantoin (97.3 fosfomycin (91.0) and chloramphenicol (78.5 were more effective than pipericillin-tazobactam (72.3 ampicillin (54.4 meropenem (45.5) and ciprofloxacin (35.7). Gentamicin and cotrimoxazole offered ineffective zones of inhibition. Only a single vre strain, also teicoplanin-resistant, was encountered.
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