A multicenter study of the antimicrobial susceptibility of Haemophilus influenzae, Streptococcus pneumoniae, and Moraxella catarrhalis isolated from patients with community-acquired lower respiratory tract infections in 1999 in Portugal

A nationwide multicenter study (including 25 laboratories) of the antimicrobial susceptibility of bacterial pathogens commonly associated with community-acquired lower respiratory tract infections (LRTI), with testing undertaken in a central laboratory, was conducted in Portugal in 1999. Antimicrobial resistance in Haemophilus influenzae has not increased in the last decade. Of the 498 isolates tested, 12.4% produced beta-lactamase and >95% were susceptible to all antimicrobials except ampicillin. In contrast, there was a rapid increase of resistance in Streptococcus pneumoniae. Of the 312 isolates tested, 24.7% exhibited decreased susceptibility to penicillin (13.5% showed low-level and 11.2% high-level resistance), 13.8% were resistant to erythromycin, clarithromycin and azithromycin, and 13.6% to cefuroxime and to tetracycline. Of the 38 Moraxella catarrhalis tested, 81.6% produced beta-lactamase. Resistance to penicillin, cefuroxime, erythromycin, clarithromycin, and azithromycin in S. pneumoniae and beta-lactamase production in H. influenzae were significantly higher in pediatric patients than in adults. Overall, amoxycillin/clavulanate was the most active antimicrobial agent in vitro against H. influenzae, S. pneumoniae, and M. catarrhalis isolated from patients with community-acquired LRTI in Portugal.     

Antimicrobial susceptibility to levofloxacin and other antibacterial agents among common respiratory pathogens—a Brazilian perspective from the GLOBAL Surveillance Initiative 2001–2002

The GLOBAL (Global Landscape On Bactericidal Activity of Levofloxacin) Surveillance programme monitored antimicrobial susceptibility patterns of the key respiratory tract pathogens Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis collected in Brazil during 1997-1998, 1999-2000 and 2001-2002. Penicillin and azithromycin resistance among S. pneumoniae strains increased from 1997-1998, reaching 7.9% and 9.5%, respectively, in 2001-2002. Although decreasing by 4.9% since the previous study, trimethoprim-sulphamethoxazole resistance remained high at 33.7%. Concurrent resistance to penicillin, azithromycin and trimethoprim-sulphamethoxazole was seen in 2.9% of the S. pneumoniae isolates collected. Levofloxacin remained extremely active against S. pneumoniae, with 0.3% resistance reported in 1997-1998 and 0% resistance in 1999-2000 and 2001-2002. beta-Lactamase production in H. influenzae was > 10% in all three studies, with correspondingly high rates of ampicillin resistance. Trimethoprim-sulphamethoxazole was the least active agent tested against H. influenzae, with resistance rates of > 40% recorded in all three studies. All H. influenzae isolates were susceptible to cefuroxime, ceftriaxone, azithromycin and levofloxacin. Of the M. catarrhalis isolates, 98.0% in 1997-1998, 98.0% in 1999-2000 and 81.8% in 2001-2002 were beta-lactamase-positive. The continued high prevalence of antimicrobial resistance in Brazil underscores the importance of current surveillance initiatives. Levofloxacin, a fluoroquinolone prescribed widely for respiratory tract infections, continued to show potent activity against key respiratory pathogens.     

Susceptibility of Enterobacteriaceae to β-lactam agents and fluoroquinolones: a 3-year survey in France

Objective   To assess trends in the susceptibility to β -lactam agents and to fluoroquinolones of clinically relevant Enterobacteriaceae isolated over a 3-year period in 14 French hospital laboratories.
Methods   During the second quarter of 1996, 1997 and 1998, 180 consecutive non-duplicate isolates of Enterobacteriaceae were collected in each center. Sixteen β -lactams and four quinolones were tested by the disk diffusion method. In addition, the double-disk synergy test was used to screen for the production of extended-spectrum β -lactamase (ESBL).
Results   Totals of 2507, 2312 and 2506 clinical isolates were obtained in each period, respectively. The distribution of Enterobacteriaceae species according to clinical specimens and wards was similar in each study period. No significant variation in the susceptibility rates to β -lactams and fluoroquinolones was observed, except in Klebsiella pneumoniae and Enterobacter aerogenes. The prevalence of ESBL-producing isolates decreased from 18% to 9% in the former, while it increased from 32% to 54% in the latter. At the same time, the susceptibility to ofloxacin and pefloxacin increased for K. pneumoniae ( P  < 0.003) and cephalosporinase-producing species ( P  < 0.05), except Enterobacter spp.
Conclusion   Over the 3-year study period β -lactams and fluoroquinolones remained highly active against Enterobacteriaceae clinical isolates, with the exception of E. aerogenes , probably as a result of the dissemination of multiresistant clones in French hospitals.     

High frequency of multiresistant respiratory tract pathogens at community level in South India

Objective: To describe the patterns of antibiotic susceptibility of outpatient strains of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis in the district of Pondicherry in South India.
Methods: The antibiotic susceptibilities of 94 S. pneumoniae , 97 H. influenzae and 104 M. catarrhalis strains, collected from outpatients with respiratory tract infections, were determined by disk diffusion and Etest.
Results: Resistance or reduced susceptibility to trimethoprim-sulfamethoxazole was found in 67% of S. pneumoniae , 53% of H. influenzae and 24% of M. catarrhalis strains. Thirty-seven per cent of S. pneumoniae and 39% of H. influenzae strains were resistant or showed reduced susceptibility to tetracycline. Reduced susceptibility to penicillin was found in 6% of S. pneumoniae strains. Overall, 10% of S. pneumoniae and 38% of H. influenzae strains showed reduced susceptibility to ≥3 antibiotics. Comparisons between the antibiotic susceptibility patterns of the Indian strains and a corresponding collection of strains from Sweden indicate that the susceptibility of the native susceptible population is independent of geographic origin.
Conclusions: The findings indicate high consumption of tetracycline and trimethoprim-sulfamethoxazole in the area, which emphasizes the need for surveillance of the pattern of antibiotic susceptibility among respiratory tract pathogens at community level in developing countries and for the implementation of local guidelines for rational use of antibiotics.     

Stability of viable-bacterium counts in liquid media used for preparation of inocula and subsequent impact on antimicrobial susceptibility test results.

We evaluated the consequences of prolonging the time between initial bacterial inoculum suspension preparation and susceptibility test inoculation. Extending the current National Committee for Clinical Laboratory Standards-recommended time of 15 min between suspension preparation and test inoculation should allow laboratories more flexibility to optimize efficiency from the standpoint of workflow. We assessed the length of time for which viable-bacterium counts remain stable in three liquid media at room temperature. Fifty isolates were examined in water, saline, and cation-supplemented Mueller-Hinton broth (CSMHB). Disk diffusion and microdilution MIC tests were performed on nine of these. Our results suggest that directly prepared inoculum suspensions of members of the family Enterobacteriaceae, Pseudomonas aeruginosa, staphylococci, and enterococci can be held for up to 6 h in water or saline prior to inoculation of disk diffusion and MIC tests without compromising test accuracy. The same organisms can be held for at least 1 h in CSMHB. Viridans group streptococci can be held for up to 6 h in saline and CSMHB and for up to 3 h in water. Similarly, Haemophilus influenzae and Streptococcus pneumoniae isolates may be held in CSMHB for up to 3 h. Because of an early decrease in viable-bacterium counts in water and saline with some H. influenzae and S. pneumoniae isolates, we recommend that National Committee for Clinical Laboratory Standards recommendations be followed for these species.     

Erythromycin susceptibility of viridans streptococci from the normal throat flora of patients treated with azithromycin or clarithromycin

Objective   To study the emergence of macrolide resistance in throat flora following treatment with clarithromycin or azithromycin.
Methods   Throat samples were collected before and after treatment and plated as a lawn on Columbia blood agar with an erythromycin E test strip. Minimum inhibitory concentrations (MICs) of erythromycin, clarithromycin and azithromycin were determined against isolates of distinct morphology with erythromycin E test MIC results equal to or greater than 2 mg/L. Polymerase chain reaction techniques were used to determine the genetic mechanisms of resistance.
Results   There were 749 resistant isolates of which 474 (63%) were streptococci. Only a quarter of the patients had no resistant streptococci before treatment started. There were increases in the numbers of resistant isolates and in the number of patients carrying a resistant flora during and after treatment. The most common genes identified were mef A/E in isolates with low-level resistance and erm A/M in isolates with high-level resistance.
Conclusions   There is a pool of streptococci carrying genes associated with macrolide resistance in the normal respiratory flora of generally healthy adults. Differences between the patients treated with clarithromycin and those treated with azithromycin were difficult to assess because of the large number of patients in each group with macrolide-resistant streptococci before treatment. Although there were some differences these were not statistically significant.     

Antimicrobial resistance in viridans group streptococci among patients with and without the diagnosis of cancer in the USA, Canada and Latin America

Objective   To investigate antimicrobial resistance in viridans group streptococci (VGS) among patients with and without the diagnosis of cancer in the USA, Canada and Latin America.
Methods   All bloodstream isolates of VGS collected from SENTRY centers in the Western Hemisphere between January 1997 and December 1999 were tested by reference broth microdilution methods (NCCLS). Results for isolates from patients with cancer were compared to those from other patient populations.
Results   Overall, 438 unique patient bloodstream isolates of VGS were collected during the study. Percentage susceptible/MIC₉₀ (mg/L) values for antimicrobials tested were as follows: penicillin, 66/1; erythromycin, 60/4; clindamycin, 92/0.12; cefepime, 86/1; trimethoprim–sulfamethoxazole, 80/2; ciprofloxacin, 44/> 2; gatifloxacin, 98/0.5; and vancomycin, 100/1. Of these isolates, 70 (16%) were confirmed to be from cancer patients. VGS isolates from cancer patients were less susceptible to most antimicrobials tested than were isolates from non-cancer patients. The greatest differences in susceptibility rates for cancer- versus non-cancer-associated VGS isolates were seen for ciprofloxacin (34% versus 46%, P  = 0.07) and trimethoprim–sulfamethoxazole (64% versus 83%, P  < 0.001), two agents which are often used for prophylaxis or as presumptive therapy in cancer patients.
Conclusions   Susceptibility rates for VGS isolates from cancer patients are lower than those for isolates from patients without a cancer diagnosis. These differences are greatest for agents that have seen widespread prophylactic and empirical use. Ongoing surveillance of VGS infections in this patient population is important and should help to guide therapy decisions.     

National surveillance programme on susceptibility patterns of respiratory pathogens in South Africa: moxifloxacin compared with eight other antimicrobial agents

AIMS: The susceptibility patterns of Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Klebsiella pneumoniae, and Streptococcus pyogenes isolated from specimens submitted to 12 private laboratories in South Africa were determined. METHODS: Minimum inhibitory concentration (MIC) determinations were performed on the isolates in the microbiology laboratory at Tygerberg Hospital according to the recommendations of the National Committee for Clinical Laboratory Standards (NCCLS). RESULTS: According to the NCCLS breakpoints, 24% of 729 S pneumoniae isolates were sensitive, 30% intermediate, and 46% resistant to penicillin. Rates of macrolide resistance were high, with 61% of the pneumococci being resistant to clarithromycin and azithromycin. Co-trimoxazole resistance was also high, with 28% of pneumococcal strains being sensitive, 21% intermediate, and 51% resistant. beta Lactamase was produced by 7% of 736 H influenzae isolates and 91% of 256 M catarrhalis isolates. The quinolones, moxifloxacin and levofloxacin, were universally active against all isolates tested, which included S pneumoniae, H influenzae, M catarrhalis, K pneumoniae, and S pyogenes. CONCLUSIONS: Haemophilus influenzae and S pneumoniae were the most commonly isolated organisms. Resistance to penicillin was one of the highest reported in the world (76%) in S pneumoniae, as was macrolide resistance in pneumonocci, although surprisingly, only 14% of S pyogenes were resistant. The quinolones, moxifloxacin and levofloxacin, were active against all organisms tested, including the penicillin and macrolide resistant strains and moxifloxacin was more active than levofloxacin against pneumococci.     

Use of the National Committee for Clinical Laboratory Standards guidelines for disk diffusion susceptibility testing in New York state laboratories

Accurate antimicrobial susceptibility testing is vital for patient care and surveillance of emerging antimicrobial resistance. The National Committee for Clinical Laboratory Standards (NCCLS) outlines generally agreed upon guidelines for reliable and reproducible results. In January 1997 we surveyed 320 laboratories participating in the New York State Clinical Evaluation Program for General Bacteriology proficiency testing. Our survey addressed compliance with NCCLS susceptibility testing guidelines for bacterial species designated a problem (Staphylococcus aureus and Enterococcus species) or fastidious (Streptococcus pneumoniae, Haemophilus influenzae, and Neisseria gonorrhoeae) organism. Specifically, we assessed compliance with guidelines for inoculum preparation, medium choice, number of disks per plate, and incubation conditions for disk diffusion tests. We also included length of incubation for S. aureus and Enterococcus species. We found overall compliance with the five characteristics listed above in 80 of 153 responding laboratories (50.6%) for S. aureus and 72 of 151 (47.7%) laboratories for Enterococcus species. The most common problem was an incubation time shortened to less than 24 h. Overall compliance with the first four characteristics was reported by 92 of 221 (41.6%) laboratories for S. pneumoniae, 49 of 163 (30.1%) laboratories for H. influenzae, and 11 of 77 (14.3%) laboratories for N. gonorrhoeae. Laboratories varied from NCCLS guidelines by placing an excess number of disks per plate. Laboratories also reported using alternative media for Enterococcus species, N. gonorrhoeae, and H. influenzae. This study demonstrates a need for education among clinical laboratories to increase compliance with NCCLS guidelines.     

The effect of carbon dioxide on susceptibility testing of azithromycin, clarithromycin and roxithromycin against clinical isolates of Streptococcus pneumoniae and Streptococcus pyogenes by broth microdilution and the Etest: Artemis Project—first-phase study

Objective: To evaluate the effect of carbon dioxide on the susceptibility testing, using broth microdilution and the Etest (AB Biodisk, Solna, Sweden), of azithromycin, clarithromycin and roxithromycin against Streptococcus pneumoniae and Streptococcus pyogenes .
Methods: Fresh clinical isolates collected from 36 hospital laboratories in 12 countries were evaluated using the Etest in the presence of carbon dioxide. The isolates were retested under ambient conditions (absence of carbon dioxide) using broth microdilution and/or the Etest.
Results: Carbon dioxide falsely elevated azithromycin, clarithromycin and roxithromycin MIC₉₀S for S. pneumoniae , determined by the Etest, approximately 12-fold. Also, the azithromycin MIC₉₀ for S. pyogenes was increased fourfold; the effect was less marked for clarithromycin and roxithromycin. When isolates were retested in the absence of carbon dioxide, using the Etest or microdilution, susceptibilities to azithroymycin were comparable to those to clarithromycin ( S. pneumoniae , 93.4% versus 91.3%; S. pyogenes , 96.4% versus 95.8%). Both organisms were less susceptible to roxithromycin ( S. pneumoniae , 71.3%; S. pyogenes , 85.7%). An internal standard control, consisting of 50 isolates each of S. pneumoniae, S. pyogenes and Haemophilus influenzae , confirmed that azithromycin susceptibility testing resulted in falsely elevated MICs.
Conclusions: Carbon dioxide falsely elevated azithromycin MICs for S. pneumoniae and S. pyogenes , with an apparent reduction in susceptibility. When the in vitro activity of azithromycin and other macrolides against S. pneumoniae and S. pyogenes is being evaluated, awareness of the pH effect is essential.     

Comparative in vitro activity of ertapenem against bacterial pathogens isolated from patients with lower respiratory tract infections

This study compared the in vitro activity of ertapenem, ceftriaxone, cefepime, ciprofloxacin and amoxicillin–clavulanate against 381 aerobic and facultative bacterial pathogens isolated from 320 patients with acute bacterial exacerbation of chronic bronchitis or community-acquired pneumonia. Streptococcus pneumoniae and Haemophilus influenzae accounted for 54.6% of the isolates. The ertapenem MIC was ≤2 mg/L for 98.4% of isolates and ≥8 mg/L for 1.0% (all methicillin-resistant Staphylococcus aureus ). Ertapenem had the most potent activity against Enterobacteriaceae, Moraxella catarrhalis , and methicillin-susceptible S. aureus , and its activity against H. influenzae and H. parainfluenzae , all strains of which were susceptible, was not altered by β -lactamase production. Only one S. pneumoniae strain, a penicillin-resistant isolate, was resistant to ertapenem. Ertapenem was highly active in vitro against pyogenic bacteria recovered from patients with community-acquired lower respiratory tract infections.     

Surveillance of linezolid resistance in Germany, 2001–2002

A surveillance study was performed throughout Germany from November 2001 to June 2002 to assess the prevalence of linezolid-resistant isolates among Gram-positive bacteria from routine susceptibility data and to compare the in-vitro activity of linezolid to that of other antibacterial agents. Each of 86 laboratories provided routine susceptibility data for 100 consecutive isolates. Most laboratories (c. 60%) used the disk diffusion test. Laboratories were also requested to send a representative sample of their isolates, as well as all isolates reported as intermediate or resistant to linezolid, to a reference laboratory for MIC determination. Susceptibility data for 8594 isolates were evaluated. Sites of infection were skin and soft tissue (29.9%), upper and lower respiratory tract (19.1%), foreign body or catheter (10.5%), or urinary tract (9.8%). Routine linezolid susceptibility data were reported for 6433 isolates. The prevalence of linezolid resistance, as reported to the clinician, was 0.4% in Staphylococcus aureus, 0.3% in Staphylococcus epidermidis, 2.9% in Enterococcus faecalis, 2.3% in Enterococcus faecium, 1.4% in Streptococcus pyogenes and 2.9% in Streptococcus agalactiae. Linezolid resistance was not detected in Streptococcus pneumoniae or in viridans group streptococci. Sixty-nine of 115 isolates reported as intermediate or resistant to linezolid were retested, but none was resistant to linezolid. Linezolid exhibited excellent in-vitro activity against representative isolates of the six most frequently encountered species (MIC90, 1-2 mg/L). The prevalence of resistance to linezolid was very low in Germany. Organisms reported as linezolid-resistant should be retested, either in the same laboratory with an alternative method or in a reference laboratory.     

P594 Intracellular killing of drug-resistant Staphylococcus aureus by different antibiotics

Objectives:  The aim of this study was to assess the carbapenem susceptibility of four nosocomial pathogens and to evaluate the reliability of the susceptibility results determined by E-test and disc diffusion (DD) methods.
Methods:  Escherichia coli ( n  = 73), Klebsiella pneumoniae ( n  = 60), Pseudomonas aeruginosa ( n  = 70) and Acinetobacter spp. ( n  = 70) isolated from nosocomial infections in 2002–2003 were included in the study. Thirty-five per cent of the strains were isolated from intensive care units. After determining antimicrobial susceptibility against imipenem and meropenem by DD (10  μ g; Oxoid, UK) and Etest (AB Biodisk, Solna, Sweden) methods, the results were categorised as susceptible (S), intermediate (I) and resistant (R) according to the NCCLS criteria. For statistical analyses, the intermediate group was included in the resistant category because of the low numbers of bacteria in the former group.
Results:  None of E. coli or K. pneumoniae strains were resistant to carbapenems, whereas, resistance reached up to 59.0% in Acinetobacter spp. and P. aeruginosa isolates. By either method, the pattern of the susceptibility of the four bacteria was not statistically significantly different for imipenem vs. meropenem. Total agreement of DD and E-test methods for susceptibility to imipenem was 95.7%, and 90.0% in Acinetobacter spp. and P. aeruginosa , respectively; and susceptibility to meropenem was 90.0% for both bacteria. However, the difference of the results obtained by either method was statistically significant for Acinetobacter spp.
Conclusion:  Study results suggest a high resistance rate for Acinetobacter spp. and P. aeruginosa strains against carbapenem antibiotics in our hospital. Further studies are needed to clarify whether E-test should be used to confirm meropenem resistance of Acinetobacter spp. determined by DD method.     

Validation and reproducibility assessment of tigecycline MIC determinations by Etest

A multicenter study was conducted to validate Etest tigecycline compared to the Clinical Laboratory Standards Institute reference broth microdilution and agar dilution methodologies. A large collection of gram-negative (n = 266) and gram-positive (n = 162) aerobic bacteria, a collection of anaerobes (n = 385), and selected collections of nonpneumococcal streptococci (n = 369), Streptococcus pneumoniae (n = 372), and Haemophilus influenzae (n = 372) were tested. Strains with reduced susceptibility to tigecycline were used with all test methods. The Etest showed excellent inter- and intralaboratory reproducibility for all organism groups tested regardless of the test methodology. The essential agreement values with the reference method (+/-1 dilution) were >99% for the collection of gram-negative and gram-positive aerobes; >98% for the S. pneumoniae, H. influenzae, and anaerobe collections; and 100% for the group of nonpneumococcal streptococci. These results validate the performance accuracy and utility of Etest tigecycline and verify the reproducibility of this convenient predefined gradient methodology for tigecycline susceptibility determination.     

Production of tumor necrosis factor-α and interleukin-6 in whole blood stimulated by live Gram-negative and Gram-positive bacteria

Objective: To investigate the production of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) induced by live Gram-negative and Gram-positive bacteria in whole blood in vitro.
Methods: In all, 49 different isolates were studied. Each of the 49 different isolates was incubated for 4 h with whole blood at a ratio of one monocyte per 1–5 bacteria. Plasma was then separated and frozen, and the concentrations of TNF-α and IL-6 were measured by enzyme immunoassays.
Results: There was a positive correlation between TNF-α and IL-6 values, r =0.9. Gram-negative bacteria induced higher levels of both TNF-α and IL-6 than Gram-positive bacteria. Group G streptococci (GGS) induced higher levels of TNF-α than Streptococcus pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis and group A streptococci (GAS). Klebsiella pneumoniae induced higher levels of TNF-α than Haemophilus influenzae, Escherichia coli and Neisseria meningitidis. GGS induced higher levels of IL-6 than Staphylococcus epidermidis, Staphylococcus aureus and GAS. When the relative amounts of cytokine induced by the strains were compared to serum concentrations measured on admission in patients with bacteremia caused by the same bacterial isolates there was no significant correlation.
Conclusion: Species- and strain-related differences in cytokine-inducing properties were found which may have significance in clinical infections.     

Changing antimicrobial susceptibility patterns among Streptococcus pneumoniae and Haemophilus influenzae from Brazil: Report from the SENTRY Antimicrobial Surveillance Program (1998-2004)

Although antimicrobial resistance rates among Streptococcus pneumoniae and Haemophilus influenzae have increased significantly in most countries in the last years, most studies from Brazil report relatively low resistance rates among these pathogens. In this study, we analyzed the susceptibility patterns of S. pneumoniae and H. influenzae from Brazil during a 7-year period. A total of 829 S. pneumoniae and 718 H. influenzae consecutively collected from 1998 to 2004, mainly from respiratory tract and bloodstream infections, were susceptibility tested by broth microdilution methods against >30 drugs and the results were analyzed by year. Overall, 77.8% of S. pneumoniae strains were considered susceptible (MIC, < or =0.06 microg/ml) to penicillin. Resistance to penicillin (MIC, > or =2 microg/ml) and ceftriaxone (MIC, > or =4 microg/ml) were detected in 7.5 and 0.5% of strains, respectively. The fluoroquinolones, levofloxacin (MIC90) 1 microg/ml) and gatifloxacin (MIC90, 0.5 microg/ml), were active against 99.8% of the isolates tested. Among the other non-beta-lactam drugs tested, the rank order of susceptibility rates was chloramphenicol (98.9%) > clindamycin (96.4%) > erythromycin (90.6%) > tetracycline (69.8%) > trimethoprim/sulfamethoxazole (36.7%). Resistance to penicillin has increased markedly among S. pneumoniae isolates over 7 years (from 2.9 to 11.0%). Additionally, resistance rates against erythromycin, clindamycin, and tetracycline decreased among pneumococcal strains during the same period. S. pneumoniae recovered from pediatric patients (< or =5 years) showed increased penicillin and trimethoprim/sulfametroxazole resistance rates compared to older populations. The rate of ampicillin resistance among H. influenzae was 14.0%, which also corresponds with the beta -lactamase production rate. All H. influenzae isolates were susceptible to amoxicillin/clavulanate (MIC90, 1 microg/ml), ceftriaxone (MIC90, < or =0.008 microg/ml), cefepime (MIC90, 0.12 microg/ml), ciprofloxacin (MIC90, < or = 0.12microg/ml), levofloxacin (MIC90, < or =0.5 microg/ml), and gatifloxacin (MIC90, < or =0.03 microg/ml). Resistance to the antimicrobials tested remained very stable among H. influenzae isolates during the 7-year study period. The continued emerging antimicrobial resistances found in these pathogens (mainly S. pneumoniae) highlight the need for alternative agents for the treatment of infections caused by these species.     

Epidemiology of penicillin resistance in Streptococcus pneumoniae isolates in Kayseri, Turkey

Objective   To determine the penicillin resistance and serotype distribution of Streptococcus pneumoniae strains and to identify clonal relationships of isolates resistant to penicillin by means of pulsed-field gel electrophoresis (PFGE).
Methods   In total, 193 S . pneumoniae strains were isolated from clinical specimens between November 1997 and January 2000. Susceptibility testing was carried out by E test, and serotyping by the Quellung reaction. Clonal relationship was analyzed by using PFGE with sma I endonuclease.
Results   Of the S. pneumoniae isolates, 23% were intermediately resistant to penicillin. There were no high-level resistant pneumococci. The majority of isolates intermediately resistant to penicillin were of serogroups/serotypes 19, 23, 14 and 1, in descending order of frequency. There were eight major clones in strains intermediately resistant to penicillin. It was seen that serogroups in the 23-valent polysaccharide vaccine, 7-valent, 9-valent, and 11-valent vaccine formulations caused 92%, 75%, 78% and 87% of pneumococcal diseases in our region, respectively.
Conclusion   Penicillin resistance in S. pneumoniae is relatively uncommon in Kayseri. All vaccine formulations can prevent the majority of pneumococcal diseases, and there is genetic heterogeneity in intermediately penicillin-resistant pneumococci in this region.     

Serotypes and clinical manifestations of group B streptococcal infections in western Sweden

Objectives   To study the serotype distributions of group B streptococci (GBS) isolated from blood and cerebrospinal fluid and from the genital tract of pregnant women and to investigate any possible relation between serotype, age and clinical manifestation.
Methods   Invasive strains were collected from 1988 to 1997 and genital strains from 1995 to 1996. Strains of GBS were serotyped with coagglutination. Clinical data were obtained from hospital notes.
Results   A total of 144 invasive strains, 78 from neonates and infants and 66 from adults, were serotyped. The most common isolates from neonates and infants were types III (62%), Ia (18%), and V (9%). The most common isolates from adults were types III (29%), Ib (23%), V (21%) and II (15%). A majority of the adults (94%) had an underlying medical condition. The most common serotypes of the 114 strains isolated from the genital tract of pregnant women were types III (32%), V (22%), Ia (13%), Ib (13%) and II (11%).
Conclusions   Serotype III was the single most frequent GBS isolate from infants and adults. Serotype V, which appeared first in 1992, was the third most frequent isolate. A vaccine containing five GBS capsular polysaccharides appears to be appropriate for the Swedish population.     

Quantitative antimicrobial susceptibility test for Streptococcus pneumoniae using inoculum supplemented with whole defibrinated sheep blood.

The National Committee for Clinical Laboratory Standards recommends the use of lysed horse blood-supplemented Mueller-Hinton broth for determining the quantitative antimicrobial susceptibility of Streptococcus pneumoniae. This procedure may be difficult for laboratories using previously prepared or commercial MIC systems. Therefore, a study was undertaken to determine whether previously prepared microdilution trays containing Mueller-Hinton broth without blood could be used for determining the antimicrobial susceptibility of S. pneumoniae by adding whole defibrinated sheep blood to the bacterial suspension used to inoculate the trays. The presence of alpha-hemolysis was used as an indicator of bacterial growth. One hundred isolates of S. pneumoniae selected to represent a distribution of susceptibility patterns were tested by the National Committee for Clinical Laboratory Standards method and the sheep blood-supplemented-inoculum method. Greater than 94% agreement between the two methods was achieved. The sheep-blood-supplemented-inoculum procedure was highly reproducible and easy to perform and provides an acceptable alternative for determining the MICs for S. pneumoniae for laboratories using previously prepared or commercial microdilution systems.     

Pneumococcal acute otitis media in children

Objectives   To evaluate the clinical and laboratory findings of Streptococcus pneumoniae acute otitis media in children during a 1 year period.
Methods   From October 1995 to September 1996, 113 children aged 2 months to 14 years (median 18 months), with S. pneumoniae acute otitis media were studied. Susceptibility testing was performed by the Kirby-Bauer method and the E- test, and serotyping by the Quellung reaction.
Results   E- test assays detected five isolates (4.4%) to be highly resistant to penicillin and 13 (11.5%) that had intermediate resistance. All isolates were found to be susceptible to vancomycin, rifampicin and cefotaxime. In total, 25 isolates (22.1%) were resistant to one or more drugs. Fifty per cent of the penicillin-resistant or intermediately resistant S. pneumoniae isolates were resistant to multiple drugs, whereas only 2.1% of the penicillin-susceptible isolates were resistant to multiple drugs. The predominating serogroups of the isolates with reduced susceptibility to penicillin were the 19 (61.1%), 9 (16.7%), 23 (11.1%), 6 (5.5%) and 14 (5.5%) whereas those of the susceptible isolates were the 19 (26.3%), 14 (13.7%), 3 (11.6%), 6 (11.6%), 9 (8.4%), 1 (5.3%) and 12 (5.3%).
Conclusions   Streptococcus pneumoniae isolates from children with acute otitis media were penicillin-insensitive in 15.9%. The multiresistant S. pneumoniae isolates belonged to serogroups: 19 (45.4%), 9 (27.3%), 6 (18.2%) and 23 (9.1%).