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N J Legakis

Publications and source records attributed to N J Legakis.

At least 91 records · Page 5Linked to original sources
PubMed1996

Variable antifungal susceptibility of wild-type Candida albicans phenotypes from neutropenic hosts.

The aim of the present study was to investigate the relationship between phenotypes of Candida albicans strains isolated from clinical specimens and susceptibility of the strains to two antifungal agents, itraconazole and fluconazole. Oropharyngeal, gastrointestinal tract, and urogenital tract specimens were collected from 131 neutropenic patients with Candida infection who had received no previous prophylactic treatment. The most frequent species isolated was Candida albicans, followed by Candida glabrata, Candida tropicalis, Candida krusei, and Candida parapsilosis. Each of the 44 Candida albicans strains recovered was found to express one of four phenotypes: smooth, irregular, fuzzy or stipple. Mean minimum inhibitory concentrations (MICs) of itraconazole and fluconazole as determined by the microdilution method and the E-test were consistently higher for Candida albicans strains expressing the stipple phenotype. The mean MICs for the four phenotypes of the Candida albicans strains ranged between 0.35 microgram/ml and 2.41 micrograms/ml for itraconazole and 2.78 micrograms/ml for fluconazole. Antifungal susceptibility of the stipple phenotype requires careful appraisal, especially in patients clinically unresponsive to azole chemotherapy or in cases of life-threatening, deep-seated Candida infections.

Analysis of Variance↗
PubMed1996

Epidemiology of community-acquired Pseudomonas aeruginosa infections in children.

The epidemiology of community-acquired Pseudomonas aeruginosa infections in children during a one-year period (January through December 1993) was evaluated. A total of 6,859 clinical samples, each one representing a separate individual with suspected infection, were cultured. Pseudomonas aeruginosa was isolated from 218 children with various infections occurring in the following order of frequency: chronic suppurative otitis media, 76.3%; appendicitis/peritonitis, 10.3%; osteomyelitis, 8.9%; skin or soft tissue infection, 6.3%; acute conjunctivitis, 3.0%; and urinary tract infection, 0.1%. A variety of O serogroups were identified: O1 (15.2%), O6 (14.7%), O11 (12.4%), O10 (11.5%), O3 (10.6%), O5 (5.1%), and O9 (4.6%). Other serogroups and nontypable strains were recovered at a frequency of 11.2% and 14.7%, respectively. Nontypable strains predominated in chronic otitis media (18.9%), while serogroups O1 (18.3%), O6 (17.5%), and O11 (17.5%) were recovered most frequently among the typable isolates. Susceptibility of Pseudomonas aeruginosa to antipseudomonadal agents was extremely high. The rate of susceptibility to ceftazidime was 99.6%, to azlocillin 98.6%, to piperacillin 98.2%, to aztreonam 97.3%, to gentamicin and netilmicin 97.7%, and to ciprofloxacin 99.1%. All isolates were susceptible to tobramycin, imipenem, and amikacin. The results might suggest that community-acquired Pseudomonas aeruginosa infections in children can be treated successfully with any antipseudomonadal antibiotic.

Age Factors↗
PubMed1996

Risk factors for nosocomial infections caused by gram-negative bacilli. The Hellenic Antibiotic Resistance Study Group.

Two hundred and ninety-nine Gram-negative hospital-acquired infections from 257 patients, consecutively identified during one month (November 1992) in five hospitals in the greater Athens area, were divided into four groups on the basis of the bacterium isolated: Group 1 (Escherichia coli group) included infections owing to E. coli, Group 2 (Proteus group) consisted of infections owing to Proteus spp. and Providencia spp., Group 3 (Kiebsiella/Enterobacter group) involved infections owing to Kiebsiella spp., Enterobacter spp., Citrobacter spp. and Serratia spp. Infections owing to Pseudomonas spp. and other non-fermenters were allocated into Group 4 (non-fermenters group). The four groups were studied in relation to risk factors including the duration of hospitalization, type of ward, underlying disease, history of operation, medical procedures/devices and antimicrobial therapy. A stepwise multiple logistic regression technique (SPSS Inc) was used to analyse the data, and the three groups (the Proteus group, the Klebsiella/Enterobacter group and the non-fermenters group) were analysed separately against the E. coli group. Infections with the Kiebsiella/Enterobacter group were associated with: (a) length of hospital stay before the infection, (b) treatment with newer antibiotics, and (c) hospitalization in an intensivecare unit (ICU). Infections with non-fermenters were associated with: (a) length of hospital stay before infection, (b) a urinary catheter, (c) type of disease (chronic infection being negatively associated), (d) treatment with newer antibiotics and (e) hospitalization in an ICU. Proteus group infections were associated with (a) length of hospital stay before infection, (b) treatment with newer antibiotics and (c) operation during present hospitalization (negative association). Interestingly, no specific hospitals were identified as risk factors. Identification of patients at risk for acquiring an infection owing to a nosocomial pathogen is vital in the development of a preventive strategy for hospital-acquired infections.

Catheterization↗
PubMed1996

Susceptibility testing of Haemophilus influenzae--an international collaborative study in quality assessment.

In order to compare the prevalence of antibiotic resistance in different geographical areas, it is necessary to ensure that agreement is achieved between laboratories on the assignment of strains to 'susceptible' and 'resistant' categories. An international quality assessment study, involving 15 laboratories in eight countries, was performed to investigate the standard of performance of the susceptibility testing of Haemophilus influenzae. One hundred and fifty strains of H. influenzae were distributed from the London Hospital Medical College (LHMC) to all laboratories who were asked to test the susceptibility of the strains to ampicillin, chloramphenicol, tetracycline, trimethoprim, cephalosporins and ciprofloxacin. Laboratories were also asked to provide the details of methodology to test the susceptibility. Significant discrepancy between the LHMC and the participating laboratories appeared in the detection of resistance to ampicillin (especially beta-lactamase-negative strains resistant to ampicillin) as well as the assignment of susceptibility and resistance to chloramphenicol, tetracycline and trimethoprim. Often these reflected the use of inappropriate breakpoints which led to erroneous assignment of susceptibility. Other variations including disc content, medium and supplement, inoculum as well as failure to measure zone sizes properly also led to some repeating anomalies.

Ampicillin↗
PubMed1996

Streptococcus agalactiae: a vaginal pathogen?

The significance of Streptococcus agalactiae as an aetiological agent in vaginitis was evaluated. A total of 6226 samples from women who presented with vaginal symptoms was examined. The presence of >10 leucocytes/high-power field (h.p.f.) was taken to be the criterion of active infection. S. agalactiae was isolated from 10.1% of these samples. The isolation rates of other common pathogens such as Candida spp., Gardnerella vaginalis and Trichomonas spp. were 54.1%, 27.2% and 4.2%, respectively, in the same group of patients. In contrast, the isolation rates of these micro-organisms in the group of patients who had no infection (<10 leucocytes/h.p.f.) were 4.2%, 38.3%, 33% and 0.5%, respectively. In the majority of samples from which S. agalactiae was isolated, it was the sole pathogen isolated (83%) and its presence was associated with an inflammatory response in 80% of patients. Furthermore, the relative risk of vaginal infection with S. agalactiae (2.38) in patients with purulent vaginal discharge was greater than that of Candida spp. infection (1.41) and lower than that of Trichomonas spp. infection (8.32). These data suggest that S. agalactiae in symptomatic women with microscopic evidence of inflammation should be considered a causative agent of vaginitis.

Adolescent↗
PubMed1995

Klebsiella pneumoniae infections in Greek hospitals. Dissemination of plasmids encoding an SHV-5 type beta-lactamase.

A total of 160 Klebsiella pneumoniae clinical strains consecutively isolated in 14 Greek hospitals in a three-month period was examined. Application of capsular typing using 72 monovalent antisera combined with phage-typing using a set of 15 Klebsiella-specific phages showed the absence of epidemic strains. However, 41% of the isolates examined displayed high level resistance to ceftazidime and aztreonam and, in most of the cases, to more than one aminoglycoside as well as to other antibacterial drugs. Nearly all of these multi-resistant strains were epidemiologically distinct on the basis of their capsular serotype and phage reactivity. After examination of 14 distinct strains, it was found that in nine cases, the resistance characters were readily transferred to Escherichia coli recipients through large self-transmissible plasmids (15-100 MDa). Six of the nine plasmids had equal molecular weight (60 MDa) and displayed similar fragment profiles upon digestion with restriction endonuclease EcoRI. Isoelectric focusing and hydrolytic studies showed that the prominent beta-lactamase produced by the transconjugants harbouring the 60 MDa plasmids and the respective K. pneumoniae parent strains, was an extended-spectrum beta-lactamase of the SHV-5 type. It appears that among K. pneumoniae strains isolated in Greek hospitals a significant resistance rate to both newer beta-lactams and amino-glycosides has been established through the acquisition of promiscuous multi-resistant plasmids which share a high degree of similarity.

Aztreonam↗
PubMed1995

Prevalent mechanisms of resistance among common enterobacterial isolates in Greek hospitals.

The recent data concerning antibiotic resistance of the enterobacteria isolated in Greek hospitals are reviewed. A variety of mechanisms of resistance, clustered in most of the cases, was observed. Epidemics of plasmids were responsible for dissemination of third-generation cephalosporins, aminoglycosides, and trimethoprim resistance among Klebsiella pneumoniae and, to a lesser extent, Escherichia coli isolates. Stable depression of the expression of chromosomal cephalosporinase is the main cause of resistance to third-generation cephalosporins observed at high frequencies in Enterobacter spp. strains.

Drug Resistance, Microbial↗
PubMed1995

Pseudomonas aeruginosa folliculitis due to non-O:11 serogroups: acquisition through use of contaminated synthetic sponges.

We describe the features of 13 cases of Pseudomonas aeruginosa folliculitis (eight of which occurred sporadically and five of which resulted from two family outbreaks) that developed subsequent to the use of commercial synthetic sponges for bathing. On the basis of O serogrouping and pyocin typing and subtyping, the strains recovered from the skin lesions were found to be identical to those isolated from household shower water and sponges. P. aeruginosa folliculitis is commonly caused by the serogroup O:11; The serogroups described in this study are rare causative agents of this type of skin infection. We believe that this is the first report of P. aeruginosa folliculitis due to serogroups O:3 and O:16.

Adult↗
PubMed1994

Alterations to the oropharyngeal and nasopharyngeal microbial flora of children after tonsillectomy and adenoidectomy.

The oropharyngeal and nasopharyngeal microbial flora of 40 children with chronically hypertrophied and infected tonsils or adenoids, was studied. Twenty of the children were treated by tonsillectomy and 20 by adenoidectomy. Swabs were taken from the surgical field pre-operatively and from the oropharynx or the nasopharynx 10 and 30 days-post-operatively. The swabs were cultured on suitable culture media and identification as well as quantitative estimation of the isolated bacteria, was carried out. Most of the potentially pathogenic bacteria exhibited beta-lactamase production. Interestingly, after surgical treatment, the respective microorganisms were quantitatively reduced and it was observed that their isolation rate was also lower. Furthermore, bacteria considered as normal inhabitants regularly showed an increase in their frequency of isolation despite the fact that a quantitative decrease was in some instances observed. These findings suggest that tonsillectomy and adenoidectomy result in an alteration to the abnormal oropharyngeal and nasopharyngeal microbial flora producing an almost normal one. It appears that, in cases of the failure of antimicrobial treatment, surgical procedures should be seriously considered.

Adenoidectomy↗
PubMed1994

Antibiotic resistance of clinical isolates of Streptococcus pneumoniae in Greece.

The antibiotic susceptibilities of 1,002 Streptococcus pneumoniae clinical isolates from patients with community-acquired pneumonia were determined over an 18-month period. Resistance rates were 14% for penicillin, 20% for erythromycin, 26% for tetracycline, and 1% for chloramphenicol. Resistance to non-beta-lactam antibiotics was associated with penicillin resistance at statistically levels.

Chloramphenicol↗
PubMed1994

Molecular epidemiology of ampicillin-resistant clinical isolates of Salmonella enteritidis.

During the last 6 years in Greece, there has been a significant increase in the number of ampicillin-resistant Salmonella clinical isolates reported. In this study 23 ampicillin-resistant Salmonella strains, consecutively isolated from patients with epidemiologically unrelated cases of food poisoning, were investigated. By serotyping and phage typing, 21 of these strains were identified as Salmonella enteritidis phage type 6a, 1 was identified as Salmonella typhimurium, and 1 was identified as Salmonella saintpaul. By plasmid pattern analysis, the 21 S. enteritidis strains were further differentiated into five groups. Group I consisted of 5 strains (carrying two plasmids of ca. 38 and 34 MDa), group II consisted of 10 strains (three plasmids of ca. 38, 34, and 2.5 MDa), group III consisted of 3 strains (four plasmids of ca. 38, 34, 15, and 2.5 MDa), group IV consisted of 1 strain (five plasmids of ca. 100, 38, 34, 24, and 15 MDa), and group V consisted of 2 strains (three plasmids of ca. 100, 38, and 24 MDa). Ampicillin resistance was easily transferred to Escherichia coli and was associated with the transfer of the 34-MDa plasmid, classified in the N incompatibility group for all strains of groups I to IV, and with the transfer of the 100-MDa plasmid for the group V strains. EcoRI restriction endonuclease digestions showed an extensive homology among the 34-MDa conjugative R plasmids. Hybridizations of the EcoRI restriction fragments of the 34-MDa plasmids with a TEM-type probe revealed the locus of the beta-lactamase gene to be situated on a ca. 6.6-MDa fragment, common in all plasmids. These results indicate that ampicillin resistance in Greece is due to the spread of a limited number of clones of S. enteritidis phage type 6A, carrying related 34-MDa R plasmids. Work is in progress to obtain a better understanding of ampicillin resistance in S. enteritidis.

Ampicillin Resistance↗
PubMed1993

Prevalence of the type I and type II DHFR genes in trimethoprim-resistant urinary isolates of Escherichia coli from Greece.

Trimethoprim resistance in 64 Escherichia coli urinary isolates from five hospitals in Greece was studied. Of the 40 isolates exhibiting transferable high-level resistance (MIC > 1024 mg/L), 21 hybridized with a specific probe for dihydrofolate reductase (DHFR) I, 13 with a probe for DHFR II, and one with a probe for DHFR V. Eleven isolates hybridized with a probe for transposon Tn7. Among the 17 isolates with non-transferable high-level resistance, seven hybridized with the probe for DHFR I, three with the probe for DHFR II, and eight were Tn7-positive. None of the seven isolates with low-level resistance (MIC 4-1024 mg/L) reacted with the probes used. Of the 28 isolates positive for DHFR I, 12 (43%) failed to hybridize with the Tn7 probe. Conversely, three isolates hybridized with the Tn7 probe, but not with the probe for DHFR I. Colony hybridization experiments showed that all but three transconjugants reacted similarly to their respective parent strains. The plasmids coding for trimethoprim-resistant DHFRs were found to differ on the basis of restriction enzyme analysis. These findings suggest that trimethoprim resistance among E. coli urinary isolates in Greece is mediated predominantly by heterogeneous transferable plasmids encoding either DHFR I or DHFR II. The dissociation between DHFR I and Tn7, together with the high incidence of trimethoprim-resistant isolates which did not hybridize with the probes for the common DHFR I or II types, indicates the continued evolution of trimethoprim resistance determinants.

Bacteriuria↗
PubMed1992

Diversity of resistance phenotypes and plasmid analysis in multi-resistant 0:12 Pseudomonas aeruginosa.

Antibiotic resistance phenotypes, plasmid content and ability of conjugal transfer of antibiotic resistance genes of 35 multi-resistant Pseudomonas aeruginosa strains were examined. The strains were isolated in 12 Greek hospitals and the majority of them (80%) belonged to serotype 0:12. The isolates were distributed to a variety of different antibiotic resistance phenotypes. Plasmid analysis showed that 10 isolates harboured plasmids ranging in size from 20 to 100 Mda. Among these strains, four carried plasmids of 100 Mda, two strains had 60 Mda plasmid each while in three strains the plasmids detected were 65, 25 and 20 Mda, respectively. One strain harboured two plasmids of 100 and 60 Mda. All strains containing plasmids belonged to 0:12 serotype, except the one harbouring the 25 Mda plasmid, which belonged to serotype 0:6. Using a P. aeruginosa recipient resistant to rifampicin and ciprofloxacin, conjugal transfer was achieved in two occasions. These plasmids, 100 Mda in size, encoded high-level resistance to both gentamicin and tobramycin whereas resistance to other drugs was not transferable. Interestingly, all 100 Mda plasmids, including the self-transferable ones, were found to share a certain degree of homology as judged by restriction analysis. It is suggested that both resistance phenotypes and analysis of plasmid content might be useful in subdividing 0:12 multi-resistant P. aeruginosa.

Drug Resistance, Microbial↗
PubMed1992

Diversity of aminoglycoside resistance in Enterobacter cloacae in Greece.

Ninety Enterobacter cloacae strains isolated from 12 Greek hospitals were examined in terms of epidemiological types and resistance mechanisms. Using O serotyping 69% of the strains were assigned to a specific serotype and overall 16 different serotypes were identified. The combination of serotyping, phagetyping and biotyping efficiently discriminated most of the strains, indicating that single epidemic strains were not prevalent, although serotypes 3, 7, and group II predominated. Eight representative strains, all resistant to gentamicin, tobramycin, amikacin and netilmicin, were further examined for transferability and mechanisms of resistance. Aminoglycoside resistance was found to be transferable in most strains, and 13 R plasmids of 40-120 MDa molecular weight were detected. The enzymes detected consisted of three enzymes active against gentamicin [ANT(2h'), AAC(3)-I and AAC(3)-V]; three active against tobramycin [ANT(2"), AAC(3)-V and AAC(6')-I]; two active against netilmicin [AAC(3)-V and AAC(6')-I]; and one active against amikacin [AAC(6')-I]. APH(3') and ANT (3"), which modify neomycin and streptomycin plus spectinomycin respectively, were also found. Overall up to five aminoglycoside modifying enzymes were detected on the same R plasmid, AAC(6')-I plus ANT(2") being the most prevalent. The high incidence of multiresistance in Enterobacter cloacae and the fact that resistance is due to enzymatic inactivation of the antibiotics, indicate that in Greece this species might act as a gene pool for the spread of resistance to other bacteria of clinical relevance.

Aminoglycosides↗
PubMed1992

Prevalence of a plasmid-mediated type II dihydrofolate reductase gene among trimethoprim-resistant urinary pathogens in Greek hospitals.

The genetic basis of trimethoprim resistance was examined in 24 Klebsiella pneumoniae, 27 Enterobacter cloacae, five Enterobacter aerogenes and nine Serratia marcescens urinary isolates from five hospitals in Greece. Analysis of the 65 isolates by serotyping and phage-typing identified 53 distinct strains. Thirty-eight isolates (15 K. pneumoniae, 19 E. cloacae, two E. aerogenes and two S. marcescens) hybridized with a probe specific for a gene encoding type II dihydrofolate reductase (DHFR). Three of the K. pneumoniae and four of the E. cloacae isolates which reacted with this probe also hybridized with probes specific for type I DHFR and transposon Tn7. Two E. cloacae isolates hybridized only with the probe for type I DHFR, while a further three isolates hybridized only with the type I DHFR and Tn7 probes. None of the isolates hybridized with a probe for type V DHFR. The plasmids in transconjugants derived from 40 isolates were analysed by digestion with restriction enzymes and Southern blotting. Eighteen (45%) of the donors (12 K. pneumoniae and 6 E. cloacae) produced transconjugants containing plasmids of about 95 kb in size, while transconjugants from the other donors had plasmids in the range 100-185 kb. Of the 18 transconjugants containing a 95 kb plasmid, 15 had similar restriction endonuclease digest patterns, although they varied in terms of the range of antimicrobial resistances which they encoded. When EcoRI digests of these 15 plasmids were hybridized with the type II DHFR probe, a 23 kb common band reacted with the probe.(ABSTRACT TRUNCATED AT 250 WORDS)

Cross Infection↗
PubMed1992

High prevalence of stably derepressed class-I beta-lactamase expression in multiresistant clinical isolates of Enterobacter cloacae from Greek hospitals.

Susceptibilities to cefotaxime (Ctx) and ceftazidime (Caz) were examined for 90 recent clinical isolates of Enterobacter cloacae from Greek hospitals. Most (68%) of the isolates were resistant to both drugs, and all were resistant to cefoxitin. beta-Lactamase activities against cephaloridine in crude extracts from Ctx-Caz-resistant isolates were high, irrespective of whether or not the cells were grown with cefoxitin as an inducer of the chromosomal beta-lactamase, indicating stable derepression of the gene for the enzyme. On the other hand, double disk antagonism tests showed that all the Ctx-Caz-sensitive isolates possessed inducible expression of this beta-lactamase. Iso-electric focusing revealed the presence of five forms of the chromosomal beta-lactamase, randomly distributed amongst the Ctx-Caz-resistant and -sensitive isolates. Plasmid-mediated beta-lactamases of TEM and PSE types also were found in many isolates. These data indicate that the extremely high prevalence of Ctx-Caz-resistant E. cloacae isolates in Greek hospitals is attributed to the dissemination of mutants which constitutively overproduce the class-I chromosomal beta-lactamase. Over 90% of these isolates exhibited cross-resistance to aminoglycosides, suggesting the accumulation of unrelated antibiotic resistance mechanisms.

Cefotaxime↗
PubMed1992

Exposure effects of netilmicin on Enterobacter cloacae clinical isolates.

The effects of netilmicin exposure on two aminoglycoside (AMG)-resistant and two AMG-susceptible Enterobacter cloacae isolates were studied. It was found that 1 h incubation of bacterial suspensions with netilmicin at 16 mg/l reduces the amounts of lipopolysaccharides present on the bacterial surface. Also, netilmicin pretreatment increased both the susceptibility to normal human serum and the adhesion to human epithelial cells of all four E. cloacae strains examined.

Bacterial Adhesion↗