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PubMed1997

Molecular epidemiology of infections with Neisseria gonorrhoeae among visitors to a sexually transmitted diseases clinic.

OBJECTIVES: To identify determinants for plasmid-mediated resistance to penicillin (penicillinase-producing Neisseria gonorrhoeae [PPNG]) and tetracycline (tetracycline-resistant N. gonorrhoeae [TRNG]) among gonococci, to determine the distribution of bacterial characteristics, and to correlate these with antibiograms and patient characteristics. STUDY DESIGN: Gonococcal isolates from 131 patients attending a sexually transmitted diseases clinic in The Netherlands in 1994 were auxotyped and serotyped and antimicrobial susceptibility was tested. Information on patient characteristics was collected at the initial visit. RESULTS: The most prevalent serotype, IB-1 (26%), proved to be related to sexual contact with casual partners, especially commercial sex partners. In addition, IB-1 strains were associated with PPNG and displayed higher minimum inhibitory concentrations (MICs) for ceftriaxone, cefuroxime, and ciprofloxacin. Homosexual men were more often infected with nonrequiring, IB-2, and IB-6 strains than heterosexuals. These strains were very sensitive to ceftriaxone and ciprofloxacin. Overall, one strain showed decreased susceptibility to ciprofloxacin (MIC 0.5 microgram/ml), but no resistance to ceftriaxone, ciprofloxacin, or cefuroxime was observed. However, 31% of the isolates were TRNG, PPNG, or both. Determinants for these resistant strains among men were the use of antibiotics (odds ratio [OR] = 4.8, 90% confidence interval [CI] 1.3-19.1), Surinam or Morrocan origin (OR = 3.3, 90% CI 1.3-8.4), and homosexual contacts (OR = 0.1, 90% CI 0.03-0.4). CONCLUSIONS: Different types, with variable susceptibility, were associated with homosexual and commercial sexual behavior. PPNG and TRNG were more commonly isolated from antibiotic users, heterosexual individuals, and ethnic minorities. Continuous surveillance of susceptibility is needed to follow the spread of PPNG and TRNG and to detect resistance to the currently recommended agents in a timely fashion.

Adolescent↗
PubMed2002

Detection of a novel Tet M determinant in tetracycline-resistant Neisseria gonorrhoeae from Uruguay, 1996-1999.

BACKGROUND: Determination of the diversity within the tet(M) sequence from N gonorrhoeae is a useful epidemiologic tool for monitoring the movement or importation of strains within a geographic region. Only two distinct tet(M) genes in clinical gonococcal isolates have been described up to now: the Dutch and the American types. GOAL: The study involved surveillance of the tet(M) gene types in high-level-tetracycline-resistant gonococcal isolates from Uruguay during the period 1996 to 1999. STUDY DESIGN: Among 181 gonococcal isolates, those showing MICs >/=16 microg/ml to tetracycline were analyzed for detection and characterization of the tet(M) gene by a polymerase chain reaction (PCR) and further HpaII restriction fragment polymorphism methods, respectively. The plasmid content and antibiogram were determined. RESULTS: Twenty-two of 181 isolates (12%) exhibited high levels of resistance to tetracycline (MICs >/=16 microg/ml) and harbored a putative 25.2-Mda plasmid that contained the tet(M) gene. A high percentage of isolates (95%; 21/22) presented the Dutch type tet(M) gene. One isolate from 1999 revealed a new restriction pattern. Such a pattern had been previously noted in 1991. This new restriction pattern has not been described previously as occurring in isolates of N gonorrhoeae. The tet(M) amplimer sequence showed 100% identity with a previously described tet(M)-carrying plasmid from N meningitidis. CONCLUSION: A new HpaII restriction pattern of the tet(M) gene is present in low frequency. The tet(M) sequence was different from the gonococcal tet(M) sequences already known and not typable with the use of a differential PCR assay. Accordingly, with the genetic diversity already present within the tet(M) sequence of N gonorrhoeae isolates, we should be aware of the sensitivity of the PCR assays in use for tetracycline-resistant N gonorrhoeae detection.

Anti-Bacterial Agents↗
PubMed2003

Ethanol-lock technique in the treatment of bloodstream infections in pediatric oncology patients with broviac catheter.

PURPOSE: To assess the ethanol-lock technique as a means of treating central venous line infections. Bloodstream infections in patients with tunneled central venous catheters can lead to removal of the lines. METHODS: Twenty-eight children and adolescents aged 2 to 18 years, with different types of cancer, had Broviac catheters and presented with positive blood culture and clinical signs of infection between January 2000 and December 2001. The ethanol-lock technique was performed 24 times in 18 patients in addition to empiric (initially) and specific (after antibiogram) intravenous antibiotic treatment. In another 15 cases, 13 children were treated with systemic antibiotics alone. RESULTS: Sixty-seven percent of the patients treated with ethanol locks had no infectious relapse of any kind within 4 weeks of treatment or during subsequent aplasia, compared with 47% treated with systemic antibiotics alone. In one boy the catheter infection could not be cleared with systemic antibiotics alone, but after one course of ethanol locks no more blood culture-positive infectious episodes were observed. No severe clinical side effects of ethanol flush were observed. Mild symptoms that occurred were tiredness, headaches, dizziness, nausea, and light-headedness. CONCLUSIONS: The ethanol-lock technique appears to be a safe, well tolerated, and effective way to treat central venous line infections, even in small children. A prospective randomized study should be designed to compare antibiotic-lock, ethanol-lock technique, and systemic antibiotics alone in the treatment of device-associated bloodstream infection.

Adolescent↗
PubMed2004

Invasive pneumococcal infections among hospitalized children in Bamako, Mali.

BACKGROUND: Prevention of invasive pneumococcal disease (IPD) in children is a global public health priority, and determination of the most common serotypes is crucial for vaccine development and implementation. METHODS: We performed prospective surveillance for IPD in hospitalized children in Bamako, Mali. All febrile children and others suspected to have invasive bacterial disease had an admission blood culture and cultures of additional anatomic sites when indicated. Standard microbiologic methods were used to identify, serotype and determine antibiograms for pneumococcal isolates. RESULTS: Of 2,049 children enrolled, 106 (5%) had an IPD, including 47 cases of meningitis and 44 bacteremic pneumonias. The incidence was highest in infants (84/100,000/year). The overall IPD case fatality rate was 24%. Only 2 of 96 isolates were nonsusceptible to penicillin. The serotypes isolated were 5 (54%), 2 (14%), 7F (10%), 19F (8%), 6A/B (3%), 9V (3%), 1 (2%) and 14 (1%). CONCLUSIONS: IPD is common and frequently fatal among hospitalized children in Mali, but surprisingly little resistance has occurred. Notably, 91% of the serotypes causing IPD in Bamako children are found in the 11-valent pneumococcal conjugate vaccine.

Adolescent↗
PubMed2005

Burden of invasive disease caused by Haemophilus influenzae type b in Bamako, Mali: impetus for routine infant immunization with conjugate vaccine.

BACKGROUND: Population-based, bacteriologically confirmed disease burden data aid decision makers in African countries pondering whether to introduce Haemophilus influenzae type b (Hib) immunization for infants. METHODS: A bacteriology laboratory was established in Hopital Gabriel Toure, serving Bamako, Mali. Children age 0-15 years with fever > or =39 degrees C or syndromes compatible with invasive bacterial disease (meningitis, etc.) were eligible. From 2 to 5 mL of blood or relevant body fluid were inoculated into Bactec Ped Plus/F medium for automated culture; body fluids were also inoculated directly onto solid media. Hib was confirmed by standard microbiologic techniques and antibiograms generated by disk diffusion. RESULTS: From June 1, 2002 to May 31, 2004, 3592 (87.8%) of 4092 children admitted to Hopital Gabriel Toure with high fever or suspected invasive bacterial disease were cultured, including 1745 who were 0-11 months old, 1132 who were 1-4 years old and 715 who were 5-15 years old. Hib was isolated from 207 Bamako children, 81 from blood alone and 124 from cerebrospinal fluid (with or without positive blood culture). Of 207 cases 204 (98.5%) occurred in children younger than age 5 years (annual incidence, 45.2/10) and 159 (77%) in infants age 0-11 months (annual incidence, 158.4/10). Peak incidence (370.0 cases/10) and 12 of 21 Hib deaths occurred in 6- to 7-month-olds. Of the Hib isolates, 11.1% were resistant to ampicillin, 32% to chloramphenicol and 0.5% to ceftriaxone. CONCLUSIONS: The substantial burden of invasive Hib disease documented in Bamako has prompted the Malian government to introduce routine infant immunization with Hib conjugate.

Adolescent↗
PubMed2000

Contamination of expressed human breast milk with an epidemic multiresistant Staphylococcus aureus clone.

Nosocomial infections caused by methicillin-resistant Staphylococcus aureus (MRSA) are a major cause of outbreaks in intensive care units. Infants make up a sector of the population that presents a high risk for MRSA infections. Mother-to-infant transmission has been indicated as a possible cause of MRSA infections in neonates. The occurrence and characteristics of MRSA in samples of banked human milk were investigated by selective culture, antibiogram and pulsed-field gel electrophoresis. MRSA contamination was found in 11% of 500 samples of expressed, fresh-frozen milk from 500 different donors at five Brazilian milk banks. The great majority of the contaminated samples passed breast milk quality control criteria for dispensing as raw milk under Brazilian and American guidelines. Most of the MRSA isolates belonged to the Brazilian epidemic clone, which is reported to be widespread in several Brazilian states, in Argentina and in Portugal. It is concluded that expressed breast milk can be a reservoir of multiresistant S. aureus epidemic clones. Studies are necessary to assess the source of contamination and potential role of MRSA-contaminated milk in the transmission of MRSA to neonates.

Anti-Bacterial Agents↗
PubMed2001

Clonal origin of aminoglycoside-resistant Citrobacter freundii isolates in a Danish county.

During 1997, attention was drawn to an increased frequency of aminoglycoside-resistant Citrobacterfreundii in a Danish county, when a total of 24 resistant C. freundii isolates was detected. In this study, 15 such isolates were typed by pulsed-field gel electrophoresis, riboprinting and partial sequencing of the gene encoding translation initiation factor 2. Fourteen of the 15 isolates were identical, as evaluated by their antibiograms and by all these typing methods. This epidemic strain harboured the aminoglycoside resistance genes aac(3)-II and ant(3")-I, with the latter located in tandem with a dihydrofolate reductase gene in a class I integron. The source of the strain remains unresolved. Representative isolates were obtained from various specimens from hospitals and general practice throughout the county, with no evidence of patient-to-patient transmission.

Aminoglycosides↗
PubMed2002

Bacillus cereus cellulitis from contaminated heroin.

Concern exists over recent unexplained deaths among intravenous drug users. This report describes a patient with crepitant cellulitis who was admitted complaining of severe pain in the right forearm. Ultrasonography demonstrated gas in the tissues and he was referred for early surgical debridement of the arm. He was treated with intravenous benzyl penicillin, gentamicin and metronidazole and made a full recovery. Aspirate samples grew Bacillus cereus, morphologically similar to the isolate obtained from a sample of the patient's own heroin. Antibiogram and API 50CHB profiles were also similar. Further typing included 'H' flagellar serotyping, which found both blood and heroin strains to be non-typable, and amplified fragment polymorphism analysis, which showed that the strains were indistinguishable. Genotyping of two selected genes from B. cereus confirmed almost certain identity between the two strains. This case illustrates the potential virulence of B. cereus when inoculated into tissues, and to our knowledge, is the first report to demonstrate a conclusive microbiological link between contaminated heroin and serious sepsis in a drug user due to B. cereus.

Adult↗
PubMed2002

A molecular and phenotypic study of Vibrio cholerae in Iran.

Vibrio cholerae is again the subject of attention on account of the current increase in the world-wide incidence of cholera. In this study, 200 clinical isolates of V. cholerae serotypes O1 and non-O1, non-O139, were collected from different provinces in Iran. The isolates were subjected to biochemical analysis, antibiogram, PCR of toxin genes, plasmid profile, ribotyping and pulsed-field gel electrophoresis (PFGE). The analysis of plasmid content showed that 33-96% of V. cholerae isolated from different provinces carry a large plasmid. PCR analysis of V. cholerae O1 showed that the genes encoding cholera toxin (ctx), toxin co-regulated pilus (tcp), accessory cholera enterotoxin (ace) and zonula occludens toxin (zot) were present in 55-97% of isolates in different provinces. Restriction fragment length polymorphism (RFLP) of BglI-digested DNA probed with five oligonucleotides revealed three different ribotype patterns in isolates of V. cholerae O1. The ribotype pattern B21 of V. cholerae O1 El Tor was found to be the predominant pattern in the isolates studied. V. cholerae non-O1, non-O139 isolates showed a single ribotype pattern. PFGE analysis also showed 10 different patterns amongst the isolates, 9 of which were in V. cholerae O1. Overall, the analysis of polymorphism of ribotypes and PFGE patterns of the isolates showed that the provinces in Iran were affected by a limited number of clones of V. cholerae O1 and non-O1, non-O139 strains.

Cholera↗
PubMed1984

Chromosomal loci associated with antibiotic hypersensitivity in pulmonary isolates of Pseudomonas aeruginosa.

492a and 492c were two strains of Pseudomonas aeruginosa isolated from the sputum of a patient with cystic fibrosis. The strains were closely related but expressed different antibiograms. 492c was hypersensitive (10-100 times more sensitive than 492a) to the beta-lactam antibiotics carbenicillin, methicillin, flucloxacillin, mecillinam and cefuroxime and the non-beta-lactam, nalidixic acid. 492c also showed enhanced sensitivity (4-8 times more sensitive than 492a) to chloramphenicol, trimethoprim and novobiocin. 492a and PAO8 expressed similar levels of antibiotic resistance, except for trimethoprim, to which 492a was five times more sensitive than PAO8. Two genes associated with antibiotic hypersensitivity were mapped in the 30 min region of the chromosome, by means of R68.45-mediated plate matings between a Leu - mutant of 492c and PAO8, followed by transductional analysis using phage F116L. The first of these genes, blsA1, was closely linked to nalB, and in a PAO background, was associated with hypersensitivity to the beta-lactams and a moderate increase in sensitivity to chloramphenicol, trimethoprim, nalidixic acid and novobiocin. A further increase in sensitivity to the latter three antibiotics was associated with the second gene, tpsA1, which mapped between ser-3 and hisV. This gene could also be transferred to PAO from 492a, thus 492c could have arisen from 492a in vivo following a single chromosomal mutation at the blsA locus. Isolation of a blsA mutant of PAO969 provided further evidence for this theory.

Anti-Bacterial Agents↗
PubMed1988

Flagella, motility and invasive virulence of Pseudomonas aeruginosa.

The role of motility as a virulence factor in Pseudomonas aeruginosa burn wound sepsis was examined using mutants deficient in the Fla or Mot phenotype. Physiological profiles of parental strains and Fla- and Mot- mutants were similar with respect to antibiograms, O antigen types, growth rates, and proteolytic, exotoxin A and phospholipase activities, providing evidence for isogenicity. Lethality studies using a subcutaneous mouse burn model showed that three Fla- mutants and one Mot- mutant were much less virulent (10(2) to 10(5) times) than the parent wild-type. Topical challenges in the flame burn model showed that a Fla- mutant of strain M-2 was approximately tenfold less virulent. A reduction in virulence, although somewhat less than tenfold, was also observed in the scald burn model for M-2 Fla-, and Mot- strains. Tissue colonization experiments revealed a characteristic, rapidly systemic infection in burned mice challenged with wild-type organisms. Nonmotile mutants similarly proliferated in the burn wound, but the characteristic bacteraemia and systemic invasion were markedly absent. The infection remained localized in the skin wound and the mice survived. The pattern of infection by nonmotile mutants in the colonization studies was very similar to that obtained with Fla+ cells in burned animals passively treated with antiflagellar antibody. These results add substantial support to the concept of motility as a P. aeruginosa virulence factor in invasive infections.

Animals↗
PubMed1979

Relationship between antibiotic resistance, the production of "virulence factors", and virulence for experimental animals in Staphylococcus aureus.

Variants that had lost some of their antibiotic-resistance determinants were selected from a multiple-antibiotic-resistant strain of Staphylococcus aureus. When tested by subcutaneous injection into guinea-pigs, and measured as the number of cocci needed to produce a skin lesion of an arbitrarily chosen diameter, the virulence of strains fell progressively with loss of resistance determinants. When the staphylococci were injected intracutaneously into mice, however, the results were less easy to interpret, but loss of resistance appeared to be associated with a reduction of the slope of the dose-response line. There was no association between the antibiogram of the strains and their production of certain enzymes and haemolysins.

Animals↗
PubMed1980

Discrimination of urinary strains of Escherichia coli by five typing methods.

A series of 156 cultures of Escherichia coli isolated from sequential urines from 20 patients with urinary-tract infection was examined by biotyping, resistotyping, haemagglutinin typing, O serotyping and antibiogram typing. From 10 of the patients, we repeatedly isolated cultures of a single strain; from each of the other 10 patients, different strains were discriminated. All cultures were typable by biotyping and resistotyping, and those techniques should prove practicable for laboratories not able to perform complete serological analysis. Only a minority (43.6%) of the cultures was typed with a limited range of 24 commercial O sera. Maximum discrimination of strains was achieved by the combined use of several of these techniques.

Adolescent↗
PubMed1984

Mechanisms of beta-lactam resistance in British isolates of Pseudomonas aeruginosa.

The mechanisms of resistance to beta-lactam antibiotics in 191 isolates of Pseudomonas aeruginosa were examined. These represented the most resistant organisms of 1866 isolates collected during a national survey of antibiotic resistance in this species. One hundred and seventy-two isolates were selected because they were resistant to carbenicillin (MIC greater than 128 mg/L) and 19 because the MICs of cefotaxime were greater than the MICs of carbenicillin. Of the carbenicillin-resistant isolates, 35 produced plasmid-mediated beta-lactamases known to be active against carbenicillin and seven produced unusual beta-lactamases; in 131 strains, resistance could not be attributed to beta-lactamase production and was considered to be intrinsic. The unusual antibiogram in which the MIC of cefotaxime was greater than the MIC of carbenicillin was associated with overproduction of the chromosomally-determined Sabath and Abrahams ' beta-lactamase. Selection of strains with this last mechanism represents a novel resistance problem and one which may increase with increased use of newer antipseudomonal beta-lactams.

Anti-Bacterial Agents↗
PubMed1986

Dissociation of surface properties and "intrinsic" resistance to beta lactams in Pseudomonas aeruginosa.

Carbenicillin resistance in strains of Pseudomonas aeruginosa isolated in Britain is mediated more frequently by "intrinsic factors" than by beta-lactamase production. Intrinsically carbenicillin-resistant isolates almost invariably were more resistant to azlocillin, cefoperazone, cefotaxime, ceftazidime, chloramphenicol, tetracycline and nalidixic acid than were carbenicillin-susceptible strains. This cross-resistance to different classes of antimicrobials suggested an impermeability-based mechanism of resistance, perhaps involving the outer membrane. The structure and composition of the outer membrane of the pseudomonas cell also influences the O-serotype specificity and the susceptibility to many bacteriophages. We therefore examined these properties for possible relationships to antibiotic resistance. Carbenicillin-resistant (122) and -sensitive (127) P. aeruginosa strains from 24 hospitals were compared. Serotype O:1, O:3, O:6, O:10 and O:11 strains predominated in both groups. Non-typable and polyagglutinating strains were infrequent in both groups. With one possible exception, none of 18 bacteriophages showed a significant preference for carbenicillin-resistant or -sensitive strains. Variation between strains was observed in the electrophoretic profile of LPS and this could be related in part to serotype, but not to antibiotic resistance. Our results contrast with those of earlier small-scale studies which have claimed relationships between surface properties and antibiogram in P. aeruginosa, and suggest that interpretation of the minor changes in LPS sometimes observed in association with the development of antibiotic resistance in vitro requires caution.

Anti-Bacterial Agents↗
PubMed1988

Fingerprinting methicillin-resistant Staphylococcus aureus by the immunoblot technique.

A series of 133 isolates of methicillin-resistant Staphylococcus aureus was fingerprinted by the immunoblot technique. Extracts were prepared by lysostaphin degradation of overnight cultures and peptides were separated by SDS polyacrylamide gel electrophoresis. The peptides were transblotted on to nitrocellulose membranes and probed with (1) a hyperimmune rabbit serum raised against a methicillin-resistant S. aureus isolate, (2) a hyperimmune rabbit serum raised against an isolate of S. epidermidis, and (3) serum from a patient who had recovered from an infection with a methicillin-resistant S. aureus. This typing method confirmed the existence of an epidemic strain that accounted for 102 of the isolates. The remaining 31 isolates were grouped into a further seven types which correlated with the results of phage typing and antibiograms.

Animals↗
PubMed1991

Epidemiological typing of coagulase-negative staphylococci by pyrolysis mass spectrometry.

Pyrolysis mass spectrometry (PMS) was evaluated for the epidemiological typing of coagulase-negative staphylococci (CNS) in situations in which it was necessary to distinguish between repeated isolation of the same strain from a single patient (genuine infection) and coincidental isolation of unrelated strains (contamination). Thirteen CNS isolates were examined, consisting of five pairs, each pair isolated from a single patient, and three unrelated strains. PMS analysis gave results equivalent to a conventional typing system comprising antibiogram, biotype and plasmid profile analysis. Both methods facilitate differentiation between genuine infection with CNS and the isolation of contaminants. The speed, reproducibility, versatility and relatively low cost of PMS suggest that it may be a valuable new technique for the epidemiological typing of CNS in routine clinical settings.

Coagulase↗
PubMed1993

Mupirocin resistance in coagulase-negative staphylococci.

High-level mupirocin resistance in coagulase-negative staphylococci isolated from patients undergoing peritoneal dialysis was investigated by transfer of the resistance determinants, usually in the form of a plasmid, to Staphylococcus aureus strains, cleavage of the plasmid by restriction endonuclease and hybridisation with a probe comprising a 4.05 kb EcoRI fragment of a plasmid from a S. aureus strain. In most instances the mupirocin-resistant staphylococci isolated from each patient were different according to the species, antibiogram and plasmid profile data. The mupirocin resistance determinant was carried on various plasmids as judged by EcoRI restriction fragment length polymorphisms. All hybridised at about 4 kb with the S. aureus probe.

Animals↗