Characteristics of Neisseria meningitidis strains isolated from patients with symptoms of meningococcal meningitis in Poland in 1995-2000

Phenotype and genotype identification of 179 Neisseria meningitidis strains isolated from cerebrospinal fluid or blood of patients with meningococcal infection, hospitalized in Poland, was performed. This is the first analysis of that type conducted in Poland. We analyzed strains collected in 1995-2000 from laboratories located all over the country. Phenotype Neisseria meningitidis B:22:P1.14 was the predominant among analyzed invasive strains in Poland. Type 22 is characteristic to most of the strains isolated in our country. No strain from analyzed group belonged to known epidemic clusters. One penicillin resistant strain (MIC = 2 mg/l) and about 27% strains with decreased susceptibility to penicillin (0.1 = < MIC < 1.0 mg/l) were present among 166 N. meningitidis tested. All strains were susceptible to ciprofloxacin and rifampicin.     

Drug resistance of Enterococcus species isolated from the urogenital system

Despite low virulence of enterococci, they have become important nosocomial pathogens. This has been correlated with the increased use of broad-spectrum antibiotics, particularly cephalosporins. Many strains of enterococci exhibit multiple drug resistance; the most important being high-level resistance (HLR) to penicillin (MIC > 100 mg/l) and gentamicin (MIC > 500 mg/l and 2000 mg/l) and/or streptomycin (MIC > 2000 mg/l). The investigation was performed on 92 strains, isolated from genito-urinary tract and recognised as Enterococcus sp. All strains were obtained from several microbiological laboratories of Gdańsk, Gdynia and Tczew. On biochemical reaction profiles species of enterococci were identified as: E. faecalis (72.8%), E. faecalis varians (9.8%), E. durans (7.6%) and E. faecium (9.8%). The minimal inhibitory concentration (MICs) of penicillin, ampicillin, azlocillin, imipenem, gentamicin, amicacin, ciprofioxacin and vancomycin were determined by the agar dilution method. None of these 92 enterococcal strains was vancomycin resistant. 22.2% of E. faecium and 7.5% of E. faecalis showed high-level resistance to penicillin. None of these strains were produced beta-lactamase. High-level resistance to streptomycin and gentamicin was detected. Both--high-level resistance to streptomycin and gentamicin--were found in 6% E. faecalis; 11.1% E. faecalis varians and 22.2% E. faecium.     

Long-term antibiotic susceptibility pattern of strains of Neisseria gonorrhoeae isolated in Czechoslovakia in the years 1957-1982

Over the past 25 years a total of 7492 strains of Neisseria gonorrhoeae have been isolated in Czechoslovakia, mainly in Prague (64%). All these strains have been tested for susceptibility to the following antibiotics: penicillin G, ampicillin, tetracycline, spectinomycin, erythromycin, doxycycline, kanamycin, rifampin, chloramphenicol, gentamicin, cephalothin, cephaloridine, lincomycin and clindamycin. In addition, seven derivatives of newer antibiotics of penicillin and cephalosporin series were tested in 1981. The study showed that in 1957 the MIC of 0.03 units of penicillin per ml was effective against 95% of strains, but in 1981 only 37% of isolates were sensitive to this concentration. The first gonococcal strains with the MIC value of 4.0 units/ml to penicillin were detected in 1981. This tendency towards decreased gonococcal susceptibility to benzylpenicillin is alarming. Over the last eight years there have been described sporadic isolations of strains relatively resistant to tetracycline (MIC = 8.0 mg/l). The susceptibility to spectinomycin has been tested in over 4000 gonococcal strains, since 1967. The test showed that this antibiotic remained highly effective against the gonococcal infection with over 95% of gonococci with the MIC value of 16.0 mg/l. No fully spectinomycin resistant strains have been found. Penicillin G as well as spectinomycin and cefotaxim are still considered the antibiotics of the first choice in the treatment of gonorrhoea. The alternative antibiotics may include cefuroxim, chloramphenicol and, in cases of sensitive strains, tetracyclines.     

Characteristics of the MRSA clone with reduced susceptibility to vancomycin

The MIC of vancomycin was determined for all S. aureus strains isolated during 1997 in one hospital. MIC values for most isolates were in the range of 0.5-2 mg/l. In 18 strains, MIC was = 6 mg/L. All these strains were MRSA. Recently described VISA strains possessed MIC values for vancomycin equal or higher than 8 mg/l and such strains were not detected in the investigated group. Although strains with MIC = 6 mg/l are not VISA, but they are candidate for reduced vancomycin susceptibility, e.g. during therapy in compromised patients. Analysis of DNA of these strains by pulsed-field gel electrophoresis (PFGE) revealed that 15 of them shared a significant similarity, allowing to place them in the same group. The comparison data of phage patterns as well as antibiotic resistance patterns strongly suggest that all these strains were derivatives of a single clone.     

In vitro activity of cefditoren against a special collection of clinical isolates of Streptococcus pneumoniae from Hungary

Cefditoren is the active form of cefditoren pivoxil, a new, broad-spectrum oral cephalosporin with strong in vitro activity against penicillin-susceptible and resistant Streptococcus pneumoniae. In this study, the minimum inhibitory concentrations (MICs) of cefditoren were determined for a special selection of S. pneumoniae isolates known to be susceptible, moderately susceptible or fully resistant to penicillin; these isolates originated from the lower respiratory tract of adults with pneumonia or the upper respiratory tract of children with or without symptoms of infection. Some of this latter group of isolates exhibited extremely high MICs to penicillin (> or = 32 mg/l), whereas the MICs of cefditoren did not exceed 2 mg/l. The MIC50 and MIC90 of cefditoren proved to be 0.25 and 1.0 mg/l, respectively, with a range of MICs < or = 0.015-2.0 mg/l for all the tested S. pneumoniae isolates. Its good activity suggests that cefditoren is expected to be a potent drug in infections caused by penicillin-resistant and multidrug-resistant S. pneumoniae.     

Antimicrobial susceptibility, β-lactamase and enterotoxin production in <Emphasis Type="Italic">Bacillus cereus</Emphasis> isolates from clinical and food samples

The antimicrobial susceptibility of 30 clinical and 30 food Bacillus cereus isolates was determined. All isolates were susceptible to streptomycin, ciprofloxacin and gentamicin, 90 % of them to clindamycin and vancomycin, and 67 % to erythromycin. All isolates were resistant to amoxicillin with clavulanic acid, ampicillin, cefotaxime, ciprofloxacin, cloxacillin, cefotaxime with clavulanic acid and penicillin. The MIC values (determined by E-tests) were 48–256 mg/L for ampicillin, 0.19–1.5 mg/L for gentamicin, 0.125–1.0 mg/L for clindamycin, 0.047–4.0 mg/L for erythromycin and 1.5–16 mg/L for vancomycin. The MICs 4.6–18.75 g/L were observed for penicillin using the microdilution method. The presence of metallo-β-lactamases was detected by E-test for 100 % of strains. Nonhemolytic diarrheal enterotoxin (NHE) was produced by 98.3 % of strains, while 31.7 % of them produced hemolytic diarrheal enterotoxin (HBL). Clinical isolates produced 10 % more HBL than food isolates. The psychrotrophic strains isolated from food samples produced NHE at 6.5 °C in 73 % of cases.     

Detection and prevalence of pneumococci with increased resistance to penicillin.

Susceptibility to penicillin was determined for 6000 strains of pneumococci isolated during 1974--76 from patients in Alberta and the adjacent region of the Northwest Territories. Strains were considered to be relatively resistant if the minimum inhibitory concentration (MIC) of penicillin was 0.16 microgram (0.26 U)/mL or more, which is eight or more times greater than the MIC for fully susceptible strains. Resistance was detected in 143 strains (2.4%) isolated from 122 patients and belonging to four capsular types. The MIC of the most resistant strains was 0.32 microgram (0.53 U/mL. Penicillin-resistant strains were highly resistant to oxacillin, the MIC being at least 30 times greater than that for penicillin-susceptible strains. Pneumococci resistant to penicillin may readily be detected by the narrowness or absence of a zone of inhibition around a 1-microgram oxacillin disc in susceptibility tests on blood agar. The degree of resistance reported here is relative and does not necessarily preclude successful treatment with full therapeutic doses of penicillin G, but penicillin preparations that give low blood concentrations may not be suitable for treating infections caused by these strains.     

Susceptibility of Neisseria meningitidis Strains from the Civilian Population to Sulfadiazine, Penicillin, and Rifampin

The minimal inhibitory concentration (MIC) values of sulfadiazine, penicillin, and rifampin for meningococcal strains isolated from civilians during 1970 were compared. The strains were isolated from various sources and geographical areas and represented several serogroups. The ranges of MIC values were as follows: 0.05 to 20 mg/100 ml for sulfadiazine, 0.01 to 0.4 mug/ml for penicillin, and 0.01 to 0.8 mug/ml for rifampin. There was no significant relationship between MIC values of sensitive or resistant sulfadiazine strains and the MIC values to the other two antimicrobial agents. Comparisons of sulfadiazine MIC values with inhibition zones around sulfathiazole discs showed excellent correlation, provided the strains were separated into sensitive and resistant groups on the basis of growth at 1 mg/100 ml. Regression curves for penicillin and rifampin sensitivity showed homologous sensitive populations with the strains studied.     

In vitro antibiotic susceptibility of Bacteroides fragilis strains isolated from excised appendix of patients with phlegmonous or gangrenous appendicitis

Out of 34 studied after-appendectomy tissues of adult and child patients 86 different strains of anaerobes were isolated. The antibiotic susceptibility of 30 isolated B. fragilis strains was tested using E tests. All studied strains were sensitive to imipenem, clindamycin and penicillin/tazobactam. Sensitivity to penicillin and cefoxitin was variable among these strains. One strain resistant to metronidazole (MIC--256 mg/L) and 3 strains with increased MIC to metronidazole were detected. Most of isolated strains were beta-lactamase producers.     

Susceptibility to macrolide antibiotics of Bordetella pertussis and Bordetella parapertussis strains isolated from whooping cough patients in 1968 and in 1997-99]

The activity of erythromycin, azithromycin, clarithromycin, dirithromycin, oleandomycin, roxithromycin, spiramycin and josamycin against 21 and 34 B. pertussis strains and against 6 and 8 B. parapertussis strains isolated respectively in the years 1968 and 1997-99 was examined. The antibiotic agar dilution method was used. The minimum concentration of macrolides which inhibited growth of B. pertussis and B. parapertussis was calculated for 50% (MIC50) and 90% (MIC90) of isolates. The susceptibility to macrolides of B. pertussis and B. parapertussis strains isolated in the years 1968 and 1997-99 did not differ significantly. The MIC90 values of erythromycin were the same for B. pertussis (MIC90 = 0.125 mg/l) and B. parapertussis strains (MIC90 = 0.25 mg/l) recovered in 1968 as for those recovered in the years 1997-99. The most active antibiotic against all strains was azithromycin (MIC90 = 0.06 mg/l). The least active antibiotics were oleandomycin (MIC90 = 2-4 mg/l) and spiramycin (MIC90 = 8 mg/l). The study showed that erythromycin remains the antibiotic of choice for treatment of whooping cough and in case of emergence of B. pertussis and/or B. parapertussis strains erythromycin resistant, can be replaced by azithromycin.     

Growth and quaternary alkaloid production in differentiating and non-differentiating strains of Ruta graveolens

Cultures of Ruta graveolens have been grown and maintained on Murashige and Skoog's medium supplemented with α-naphthalene acetic acid (1 mg/l.) and kinetin (0.1 mg/l.) or 2,4-dichlorophenoxyacetic acid (1 mg/l.), and glucose (30 g/l.). The production of dihydrofuroquinoline alkaloids has been investigated in these cultures. The present study showed that: (a) cultures of Ruta produced both differentiating and non-differentiating strains; (b) quaternary alkaloids (platydesminium, ribalinium and rutalinium) were present in all the strains and maximum contents were detected at 3-4 weeks growth; (c) high platydesminium and rutalinium content was associated with differentiation and callus formation, respectively; (d) changed concentrations of quaternary alkaloids were recorded in all the five strains grown on media supplemented with tryptophan and precursors (anthranilic acid, 5-methylanthranilic acid and 2,4-dihydroxyquinoline).     

Prevalence and chemosusceptibility of Neisseria meningitidis and Haemophilus influenzae in a population of central Italy

The purpose of this study was to determine the prevalence of nasopharyngeal carriers of Neisseria meningitidis and Haemophilus influenzae, and to evaluate their chemoresistance. N. meningitidis was more frequently isolated in adolescents (10-15 years). All of meningococci were susceptible to ceftriaxone and rifampin, only one isolate showed reduced susceptibility to chloramphenicol and four strains showed reduced penicillin susceptibility. The results show that these drugs are still effective for prophylaxis and treatment in our area. All strains of H. influenzae were susceptible to penicillin, ceftriaxone, chloramphenicol, rifampin, azithromicin and gentamicin. 6 nontypeable strains were resistant to sulfamethoxazole-trimethoprim, 7 strains of type a and c-f, and 3 non-typeable strains showed reduced susceptibility to tetracycline. In contrast with the current trend in the world, in our area the susceptibility of H. influenzae to betalactams was 100%, therefore these antibiotics are still the drugs of choice for treatment of invasive diseases.     

Antibiotic sensitivities of Neisseria gonorrhoeae in the Toronto area

The antibiotic sensitivity pattern of 3872 isolates of N. gonorrhoeae tested in Toronto from 1969 to 1973 is reviewed. An increase in resistance to both penicillin and tetracycline was noted up to 1971, but no further increase has occurred since then. Ninety-seven percent of 135 patients with “sensitive” strains (inhibited by 0.3 U/ml of penicillin and/or 0.5 μg/ml of tetracycline) were cured by either 8 g of tetracycline or 5,000,000 U of penicillin, whereas only 59% of 58 patients with “resistant” strains (requiring 1.0 U/ml of penicillin and/or 2.0 μg/ml of tetracycline for inhibition) were cured by the same dosage. Spectinomycin appears to be an acceptable alternative therapy. Maximum doses of the chosen drug are recommended in the hope of retarding further spread of more resistant organisms.     

Production and purification of statins from Aspergillus terreus strains

Lovastatin, mevastatin, pravastatin and monacolin J were produced using Aspergillus terreus strains. Mevastatin (170 mg/l) was obtained at 14 days from the A1 strain, lovastatin (256 mg/l) at 21 days from the A2 strain and pravastatin (270-300 mg/l) at 14 days from both the A1 and A2 strains grown on defatted soybean flour. Similar yields of monacolin J (5-10 mg/l) were detected for both strains. Fermentation carried out by adding glycerol to A1 7-d old cultures gave 244 mg lovastatin/l at 14 days employing whole soybean flour. A new extraction procedure was applied to an A2 19-d old culture on the mycelium and the culture filtrate separately. Recovery yield showed that 83% lovastatin was associated with the mycelium and 17% was free in the culture filtrate. © Rapid Science Ltd. 1998     

Antibiotics and the Oral Streptococci of Man

The effects of 3 antibiotics, phenoxymethylpenicillin, cephalexin and clindamycin on the normal oral streptococcal flora in the region of the gingival crevice were investigated because these organisms are able to cause subacute bacterial endocarditis. Secretion of these antibiotics into the oral cavity was also examined. Penicillin and clindamycin exerted marked effects on the normal oral streptococci, whereas cephalexin did not cause any obvious change in the total flora. Following penicillin therapy, streptococci resistant to 1.5 μg/ml penicillin were observed and these organisms could be detected at least 8 weeks after the last dose of the antibiotic. They probably arose by selection from the mixed flora. Following cephalexin therapy, a much lower proportion of streptococci resistant to 15 μg/ml was found. The proportion of resistant strains fluctuated appreciably, however, probably due to their transient nature. Streptococci resistant to 1 μg/ml clindamycin were not observed in 10 out of 11 treated subjects.
Penicillin and clindamycin could be detected in the pooled saliva and gingival fluid after administering single doses of 500 mg and 300 mg, respectively. The peak levels were obtained between half and 1 h. The concentration of penicillin dropped rapidly within 3 h but clindamycin could be detected at significant levels for at least 6 h. No cephalexin could be detected in the pooled saliva or gingival fluid after a 500 mg dose. The implications of these findings in the prevention of subacute bacterial endocarditis are discussed.     

Evaluation of Argentinean white rot fungi for their ability to produce lignin-modifying enzymes and decolorize industrial dyes

The decolorizing capacity of 26 white rot fungi from Argentina was investigated. Extracellular production of ligninolytic enzymes by mycelium growing on solid malt extract/glucose medium supplemented with different dyes (Malachite Green, Azure B, Poly R-478, Anthraquinone Blue, Congo Red and Xylidine), dye decolorization and the relationship between these two processes were studied. Only ten strains decolorized all the dyes, all ten strains produced laccase, lignin peroxidase and manganese peroxidase on solid medium. However, six of the strains could not decolorize any of the dyes; all six strains tested negative for lignin peroxidase, and produced less than 0.05 U/g agar of manganese peroxidase. Comparing the isolates with the well-known dye-degrader Phanerochaete chrysosporium, a new fungus was identified: Coriolus versicolor f. antarcticus, potentially a candidate for use in biodecoloration processes. Eighteen day-old cultures of this fungus were able to decolorize in an hour 28%, 30%, 43%, 88% and 98% of Xylidine (24 mg/l), Poly R-478 (75 mg/l), Remazol Brilliant Blue R (9 mg/l), Malachite Green (6 mg/l) and Indigo Carmine (23 mg/l), respectively. Laccase activity was 0.13 U/ml, but neither lignin peroxidase nor manganese peroxidase were detected in the extracellular fluids for that day of incubation.     

Bacteriostatic or bactericidal effect of linezolid against multiresistant Streptococcus pneumoniae

The nasopharynx plays a critical role as the reservoir of Streptococcus pneumoniae, including drug-resistant strains particularly in children attending day care centers. A total 58 nasopharyngeal, multiresistant isolates of S. pneumoniae collected from healthy pre-school children were susceptible to linezolid (MIC = 0.25-1 mg/l), irrespective of serotype and drug resistance pattern. The majority of them (about 94%) were sensitive to the bactericidal effect of linezolid with MBCs = 0.5-4 mg/l. One isolate was killed at 8 mg/l of linezolid, while two at higher concentrations of this antibiotic with MBCs 16 or 32 mg/l, suggesting tolerance of linezolid. BOX-PCR fingerprinting data imply that two linezolid-tolerant strains belonged to distinct clones. Linezolid tolerance was confirmed by monitoring the viability of these isolates during exposure to 4 or 20 mg/l of this antibiotic. The linezolid-tolerant strains were sensitive to the bactericidal effect of vancomycin. Linezolid tolerance in clinical isolates of pneumococci may represent a potential therapeutic risk, especially in infections in which bactericidal activity of drug is critical for eradication of bacteria.     

Efficacy and safety of azithromycin prophylaxis of respiratory tract infections in military community]

The efficacy and safety of azithromycin prophylaxis of community-acquired pneumonia (CAP) in young adults in a military training centre of the Ministry of Defence of the Russian Federation located in the Central European Region of Russia were studied. Two prophylactic regimens with azithromycin vs. the control were evaluated: azithromycin, 500 mg/w for 8 weeks (R1), azithromycin, 1500 mg once upon the enrolment (R2) and no drugs (R3). Nasopharyngeal carriage of Streptococcuspneumoniae and its susceptibility to antibacterials were estimated thrice: before the exposure, after the exposure within the 9th week and after the exposure within the 20th week. The MLS(B) phenotype was suspected when the isolates were resistant to erythromycin and clindamycin. During the observation period of 22 weeks CAP was diagnosed in 20.2% of 678 subjects in group R3, 8.6% of 508 subjects in group R1 (Risk Ratio =0.4, 95% Cl = 0.3-0.6) and 10.3% of 507 subjects in group R2 (Risk Ratio = 0.5, 95% Cl = 0.4-0.7). The S.pneumoniae carriage rate at visit 0 was 34-35%, within the 9th week it was 75, 66 and 50% (p<0.05) in groups R1, R2 and R3 respectively, and within the 20th week it was 69, 57 and 36% in the same groups (p<0.05). At visit 0 no macrolide resistance was detected in any of the 40 isolates tested. The background level of intermediate penicillin resistance was revealed in 0-14% of the isolates. Dramatic growth of macrolide resistance was observed within the 9th week in group R1 (95.7%, 44 resistant strains, Azithro+Clinda resistance in 37% of them) and in group R2 (89.5%, 34 resistant strains, Azithro+Clinda resistance in 11.9% of them). By the 20th week the resistance rate decreased up to 40 % (16 resistant strains, Azithro+Clinda resistance in 10% of them) in group R1 and up to 22.6% (7 resistant strains, Azithro+Clinda resistance in 5.4% of them) in group R2. As for penicillin resistance, no unfavourable shifts were detected. The study demonstrated the effectiveness of the azithromycin prophylaxis of CAP in healthy young men at high transient risk of the disease, as well as the possible risk for selection of resistant endemic pathogens.     

Effects of penicillin G on morphology and certain physiological parameters of Lactobacillus acidophilus ATCC 4356

Evidence shows that probiotic bacteria can undergo substantial structural and morphological changes in response to environmental stresses, including antibiotics. Therefore, this study investigated the effects of penicillin G (0.015, 0.03, and 0.06 mg/l) on the morphology and adhesion of Lactobacillus acidophilus ATCC 4356, including the colony morphotype, biofilm production, hydrophobicity, H?O? formation, S-layer structure, and slpA gene expression. Whereas only smooth colonies grew in the presence of penicillin, rough and smooth colony types were observed in the control group. L. acidophilus ATCC 4356 was found to be hydrophobic under normal conditions, yet its hydrophobicity decreased in the presence of the antibiotic. No biofilm was produced by the bacterium, despite testing a variety of different culture conditions; however, treatment with penicillin G (0.015-0.06 mg/l) significantly decreased its production of H?O? formation and altered the S-layer protein structure and slpA gene expression. The S-protein expression decreased with 0.015 mg/l penicillin G, yet increased with 0.03 and 0.06 mg/l penicillin G. In addition, the slpA gene expression decreased in the presence of 0.015 mg/l of the antibiotic. In conclusion, penicillin G was able to alter the S-layer protein production, slpA gene expression, and certain physicochemical properties of Lactobacillus acidophilus ATCC 4356.     

Novel gyrase mutations and characterization of ciprofloxacin-resistant clinical strains of Enterococcus faecalis isolated in Poland

The activity of ciprofloxacin, sparfloxacin and moxifloxacin was determined for 205 Enterococcus faecalis isolates from patients of five hospitals (Warsaw, Poland; collected from 2000 to 2002). Ciprofloxacin resistant and intermediate isolates were numerous (53.7%). Among them, highly resistant (MIC > or = 16 mg/l) isolates predominated (98%). Isolates resistant to ciprofloxacin were also resistant to sparfloxacin and moxifloxacin. The parC and gyrA QRDRs (quinolone-resistance-determining region) of 11 isolates with ciprofloxacin MICs from 1 to 256 mg/l were analysed by DNA sequencing. In ParC one kind of amino acid substitution (of Ser-85 to Ile) in 9 E. faecalis strains with MICs from 16 to 256 mg/l was observed. In GyrA Ser-84 was changed to one of four different amino acids: Arg, Ile, Cys or Tyr, however no association between the amino acid type and MIC value was found. The last two substitutions have not been reported to date for E. faecalis. Moreover, our results may suggest that mutations within parC and gyrA are associated with development of a high-level of ciprofloxacin resistance.