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Synergistic activity of aminoglycoside-beta-lactam combinations against Pseudomonas aeruginosa with an unusual aminoglycoside antibiogram.

The bactericidal activity of aminoglycosides alone and in combination with various beta-lactams was studied by the time-kill technique against ten Pseudomonas aeruginosa isolates with an unusual antibiogram (amikacin-resistant, gentamicin-resistant, tobramycin-susceptible [ArGrTs]). Previous studies have indicated that ArGrTs isolates are moderately resistant to all aminoglycosides and many are multiply resistant to beta-lactams. Aminoglycoside-beta-lactam combinations showed infrequent synergistic (16%) or enhanced killing (12%) against the ArGrTs isolates. Synergistic activity, when present, was more likely to occur with tobramycin and amikacin than with gentamicin, even though these differences were not statistically significant. Antibiotic resistance patterns were not predictive of synergy or enhanced killing. Systemic infections produced by ArGrTs isolates that are multiply resistant to the beta-lactams may not respond to combination therapy with an aminoglycoside and beta-lactam. Alternative treatment with polymyxin B or a quinolone may be required for these infections.

Aminoglycosides↗

Staphylococci of the normal human skin flora. Variety of biotypes and antibiograms without direct correlations.

352 strains of Staphylococci of the normal human skin flora were sampled from one volunteer by single scrabbing in a ca. 3 cm2 measuring area. They were biotyped by the scheme of Pelzer et al.(1973)--a modified Baird-Parker-Scheme (1963)--and the resistance to antibiotics was investigated by the method of Bauer et al. (1966). All the nine biotypes of Staphylococci were found in variable quantities. It seems problematic to call one biotype as the main type. Morphologically identical colonies of Staphylococci from the indigenous flora of the human skin were not identical in their biotypes as previously described by Pelzer (1976). Only the investigation of all Staphylococci colonies from the culture plate can evaluate all biotypes of Staphylococci of the normal human skin flora, and can give the right quantitative correlation. Staphylococci were found to be sensitive and resistant up to four antibiotics, and one biotype did not show one type of antibiogram.

Adult↗

Multivariate analysis of antibiograms for typing Pseudomonas aeruginosa.

A method for typing Pseudomonas aeruginosa using antibiotic susceptibility patterns is presented, which allows recognition of clusters of the same strain among clinical isolates from different patients, thus indicating whether cross infection has occurred. An index of similarity (the euclidean or the oblique distance), which includes all the differences of disk zone sizes among isolates, is computed and then elaborated by a clustering algorithm that successively groups all the isolates in larger clusters. The results of clustering are presented as dendrograms, whose terminal branches are pruned down to a level below which differences are casual; isolates that still appear on a common branch are considered identical. The reliability of this technique for detecting nosocomial cross infections was assessed by comparing its results with that of serotyping and pyocin typing. Only 2 of 31 (6.4%) clusters detected by multivariate analysis were not confirmed, while 4 of 33 (12.1%) clusters were recognized by serotyping and pyocin typing, but not by multivariate analysis. In at least two instances the differences in susceptibility patterns were due to cytoplasmic R factors. The routine use of antibiogram data for typing purposes should be considered an essential part of nosocomial infection control.

Anti-Bacterial Agents↗

Development of antibiotic resistance by staphylococcus aureus in a single patient. Confirmation by phage typing, antibiograms, and plasmid analysis.

A patient is described who had colonization and infection with strains of Staphylococcus aureus increasingly resistant to multiple antibiotics while receiving several courses of broad-spectrum antibiotics. S. aureus strains with different phage types, antibiograms, and plasmid profiles were isolated from different sites at the same time. After the patient was transferred to an intermediate-care service where he received only supportive care and no further antibiotics, culture samples from areas previously colonized or infected (sputum, anterior nares, perineal region, rectum) showed no S. aureus. This patient graphically demonstrates that antibiotics may create an environment allowing the overgrowth of resistant organisms and that withdrawal of this selection pressure may allow more sensitive organisms to recolonize.

Adult↗

[Interpretive reading of the antibiogram of enterobacteria].

Many of the resistance mechanisms of enterobacteria to antimicrobial agents are well understood; nevertheless several aspects remain unsolved, particularly with regard to prediction of clinical response. The resistance pattern observed in the antibiogram of a specific organism should be the sum of the natural resistance pattern, characteristic of the species, plus the acquired resistance. In enterobacteria the principal mechanism of resistance to beta lactams and aminoglycosides is enzyme production, Each enzyme recognizes one or more specific beta lactam or aminoglycoside, as a substrate. This translates as a specific resistance phenotype that allows one to infer the enzyme(s) implicated. Enzyme resistance is not, however, the only mechanism of resistance to these agents; often the pattern observed is multifactorial. Resistance to quinolones is due to point and sequence mutations which may be selected by initially active fluoroquinolones and cause a stepwise increase of resistance.

Acetyltransferases↗

[Interpretative reading of the non-fermenting gram-negative bacilli antibiogram].

Among non-fermenting Gram-negative rods, the most clinically important species are Pseudomonas aeruginosa, Acinetobacter baumannii, and Stenotrophomonas maltophilia, which are frequently multiresistant. P. aeruginosa resistance to beta-lactams depends on the production of chromosomal and plasmid-mediated beta-lactamases, altered permeability (loss of OprD porin is related to carbapenem-resistance) and active efflux pumps, particularly MexAB-OprM. In aminoglycoside-resistant strains the main mechanism of resistance is the production of inactivating enzymes; the efflux pump MexXY-OprM is also involved. Quinolone-resistance in P. aeruginosa is related to changes in topoisomerases, altered permeability and efflux pumps. The mechanisms of resistance of A. baumannii have not been well characterized, which makes interpretative reading of the antibiogram in this organism difficult. Resistance to beta-lactams is associated with the production of beta-lactamases and altered penicillin-binding proteins. Resistance to aminoglycosides has been related to modifying enzymes and resistance to quinolones to altered targets. S. maltophilia is resistant to carbapenems and other beta-lactams because of the production of two beta-lactamases (L-1 and L-2). Aminoglycoside-modifying enzymes have also been described in this species. In contrast to what is observed in other organisms, S. maltophilia resistance to quinolones has been mainly related to active efflux, rather than to target alterations.

Acinetobacter↗

Phage susceptibility, enterotoxigenicity and antibiograms of Staphylococcus aureus strains isolated from human wounds and diarrhoea.

The phage types, enterotoxigenicity and antibiograms of Staphylococcus aureus strains isolated from human diarrhoea and skin wounds in Nigeria were determined. Of 194 strains tested, 140 (72.2%) were typable using a combination of phages in the International Phage Set (IPS) for human strains of S. aureus and the bovine phage set. IPS phages lysed 118 (60.8%) strains while 96 (49.5%) were susceptible to bovine phages. S. aureus strains from adult diarrhoea were significantly (P less than or equal to 0.01; x2) more sensitive to bovine phages (52.8%) than to IPS human phages (16.7%). Strains isolated from wounds were however significantly (P less than or equal to 0.001; x2) more susceptible to IPS human phages (72.9%) than to bovine phages (41.5%). Phage group III strains were predominant amongst diarrhoeal isolates while wound strains were most susceptible to group II phages. Phage 119, a bovine phage, lysed all 19 strains of diarrhoeal origin sensitive to group M phages but all wound strains were resistant. Overall, 132 (68.0%) strains were enterotoxigenic producing staphylococcal enterotoxins A (SEA), B (SEB), C (SEC) or a combination of these. A majority of diarrhoeal strains elaborated SEC while SEB production was predominant amongst wound strains. Of the eight antimicrobial agents used, S. aureus strains were most resistant to penicillin (71.6%) and least to sulfamethoxazole/trimethoprim (1.0%). Occurrence of resistance to one or more antibiotics was higher amongst wound strains (97.5%) than amongst strains isolated from diarrhoea (52.6%). It was concluded that bovine phage 119 may be a useful epidemiologic marker for S. aureus strains of bovine origin associated with human diarrhoea in Nigeria. It is however difficult to ascribe any aetiological significance to these strains because other enteropathogens not assayed for may have been present.

Adult↗

Antibiogram pattern and seasonality of Salmonella serotypes in a North Indian tertiary care hospital.

The antibiogram pattern and seasonal distribution of Salmonella serotypes were analysed retrospectively over a 6-year period from January 1999 to December 2004. Blood cultures received in the Bacteriology Laboratory were processed by standard procedures and the Salmonella spp. isolates were identified with specific antisera and standard biochemical tests. Antimicrobial susceptibility testing was carried out by a standard disc diffusion method and the minimum inhibitory concentration (MIC) of ciprofloxacin for 332 representative Salmonella isolates was determined by E test. Salmonella Typhi (75.7%) was the predominant serotype among 830 Salmonella spp. isolated during the study period followed by S. Paratyphi A (23.8%). The maximum number of enteric fever cases occurred during April-June (dry season) followed by July-September (monsoon season). There was a decrease in multidrug-resistant (MDR) S. Typhi, but MDR S. Paratyphi A isolates increased. There was also a dramatic increase in nalidixic acid-resistant isolates. All isolates were susceptible to third-generation cephalosporins and ciprofloxacin except one S. Typhi strain which demonstrated high-level ciprofloxacin resistance with a MIC of 16 mug/ml. A knowledge of the seasonal distribution and antibiotic resistance pattern of Salmonella in a particular geographical region is helpful in the delineation of appropriate control measures required for prevention of enteric fever.

Adult↗

Prevalence and antibiogram of Campylobacter jejuni in domestic animals in rural Ghana.

A total of 134 samples of rectal and cloacal swabs taken from apparently healthy domestic animals that were in 43 of 76 homes located in rural Ghana were examined for Campylobacter jejuni. C. jejuni was isolated from 32.8% (44/134) of the animals. The highest isolation rate of 43.6% was from local domestic fowls, followed by goats (33.3%) and sheep (23%). This organism was not detected in the pigs, cats, and ducks. All 44 strains of this bacterium produced hydrogen sulphide and hydrolyzed hippurate; antibiogram on 24 strains showed resistance to cephalothin, cephalexin, and sulfamethoxazole/trimethoprim. C. jejuni biotype 2 is the prevailing strain in animals in the area studied.

Animals↗

Microbial isolates and antibiogram from endocervical swabs of patients with pelvic inflammatory disease.

The study was aimed at determining the microorganisms isolated commonly from endocervical swab cultures of patients with suspected pelvic inflammatory disease and their in vivo antibiotic sensitivity patterns. All records of microbial culture and antibiotic sensitivity test results of suspected cases of pelvic inflammatory disease at the Federal Medical Centre Gombe from January to June 2001 were retrieved and analysed using the SPSS statistical package. Four hundred and thirteen endocervical swabs were cultured, with bacteria and/or fungal pathogens isolated in 312 (75.5%) cases. There were seven microbial organisms isolated with Staphylococcus aureus as the most commonly isolated organism in 45% (142/312) of cases. Mixed growth occurred in 46 cases (11.1% of cases), mostly bacterio-fungal 34/46 (73.9%). Only seven antibiotic disks were used in testing for sensitivity of the isolated bacteria. Cephalexin and gentamicin were the only antibiotics with a >50% sensitivity rate of 88% and 56.9%, respectively. Co-trimoxazole, penicillin and tetracycline were the most frequently resistant antibiotics with 98.4%, 97.6% and 90.8% resistance rates, respectively. It is clear that the facilities both for isolating organisms and testing for their antibiotic sensitivity pattern are limited in developing countries, despite the well-known complications associated with pelvic inflammatory disease. This would make empirical antibiotic chemotherapy common, thus confounding further the problem of management. This may be overcome by establishing well-manned and equipped regional centres that could advise other centres of trends in pattern of isolates and antibiograms.

Bacterial Infections↗

Antimicrobial susceptibility patterns (antibiograms) as an aid in differentiating Citrobacter species.

The hydrogen sulfide-negative Citrobacter group represents a taxonomic problem. Various investigators have proposed such designations as Padlewskia, Levinea, atypical Enterobacter cloacae, H2S-negative variants of Citrobacter, Citrobacter koseri and Citrobacter diversus. This problem has been investigated with emphasis on antibiograms as a means of discrimination. Clinical isolates fitting the designation Citrobacter were studied and, using the criteria of Ewing and Davis, separated into two groups: C. diversus (40 strains) or C. freundii (25 strains). Susceptibilities to ampicillin, carbenicillin, cefazolin, cephaloridine and cephalothin were determined by the agar-dilution method. C. diversus strains were resistant to 8 mug/ml ampicillin (97.5%) and 32 mug/ml carbenicillin (87.5%), and were susceptible to 8 mug/ml cephalosporins (greater than or equal to 90%). C. freundii strains were moderately susceptible to 8 mug/ml ampicillin (25%) and susceptible to 8 mug/ml carbenicillin (92%), and were resistant to 8 mug/ml cephalosporin (greater than or equal 92%). Using these data one can separate C. diversus from C. freundii with 90% accuracy.

Ampicillin↗

Antibiogram-resistogram typing scheme for methicillin-resistant Staphylococcus aureus.

An antibiogram-resistogram (AR) typing scheme that can simply and rapidly differentiate methicillin-resistant Staphylococcus aureus (MRSA) isolates has been devised. Susceptibility to antibiotics and chemicals was determined by disk diffusion testing. Three disk diffusion methods and three control S. aureus strains were evaluated. A modified Stokes' technique in which S. aureus ATCC 25923 replaced S. aureus NCTC 6571 as the control organism was chosen. Susceptibility patterns against 18 antibiotics and four chemicals were used to determine AR types. AR subtypes were determined with reference to knowledge of the local MRSA population so that plasmid loss would not result in misclassification. AR typing was compared with phage typing and plasmid profiling and found to be more discriminatory than either of these typing methods. Representative isolates of the most frequently occurring AR patterns were further characterised by investigating enterotoxin production, MICs of gentamicin and amikacin, and restriction endonuclease analysis of plasma DNA, to determine whether apparently different strains could have the same AR pattern and to devise confirmatory tests for any such similar patterns. One pattern was shared by two strains but isolates could be differentiated by susceptibility to minocycline. This typing scheme can be used in the diagnostic laboratory and results may be obtained within 24 h.

Anti-Bacterial Agents↗

Evaluation of an antibiogram-resistogram typing scheme for methicillin-resistant Staphylococcus aureus.

Between Dec. 1992 and Aug. 1993, the MRSA population in the Federated Dublin Voluntary Hospitals and St James's Hospital group was studied with an antibiogram-resistogram (AR) typing scheme in which AR patterns were determined by testing susceptibility to 22 antibiotics and chemicals by a modified Stokes' disk diffusion technique. The typing scheme divided this MRSA population into 31 AR types but 90% of isolates belonged to seven types. Isolates belonging to the most frequently occurring types (AR types 13 and 14) differed only in their reaction to lincomycin (or clindamycin) and could not be distinguished by phage typing, plasmid profiling or restriction endonuclease analysis. The AR typing scheme showed that the incidence of different AR types varied in different hospitals and changed during the study period. This typing method differentiated a strain of MRSA responsible for a nosocomial outbreak in an intensive care unit from other MRSA isolated in the unit, and has distinguished imported strains from local ones. In one hospital, AR typing showed that, although a major outbreak occurred with one AR type, there was also a series of smaller outbreaks with other AR types. The technique can be performed in the diagnostic laboratory and results were available within 24 h.

Anti-Bacterial Agents↗

Salmonella typhimurium DT 193: differentiation of an epidemic phage type by antibiogram, plasmid profile, plasmid fingerprint and salmonella plasmid virulence (spv) gene probe.

Of over 2000 isolates of Salmonella typhimurium DT 193 from humans examined in the 2 year period 1991-92, 93% were antibiotic-resistant with the most common R-types being ASSuT (38%) and T (29%). Fourteen plasmid profiles were identified in DT 193 R-type ASSuT with the majority of isolates being characterized by a single plasmid of 80 MDa (pDEP 34) which in addition to coding for ASSuT, also hybridized with a spv gene probe prepared from the 50 MDa Salm. dublin serovar-specific plasmid. On the basis of restriction fragment length polymorphisms, two variant lines of pDEP 34-like plasmids were identified and a third line which had lost the genes coding for resistance to ampicillin, streptomycin and sulphonamides, was recognized. Although 18 plasmid profile types were identified in DT 193 R-type T, all isolates carried a high mol. wt plasmid which coded for tetracycline resistance only. Further discrimination was achieved on the basis of hybridization of tetracycline resistance plasmids with the spv gene probe and restriction enzyme fingerprinting. These results demonstrate that Salm. typhimurium DT 193 can be rapidly subdivided by antibiogram and that further subdivision can be achieved on the basis of plasmid profile, plasmid fingerprint and hybridization with a spv gene probe.

Animals↗

Pseudomonas aeruginosa isolates recovered from clinical and environmental samples in Buea, Cameroon: current status on biotyping and antibiogram.

OBJECTIVE: The study was aimed at determining the prevalent biotypes of Pseudomonas aeruginosa in the environment of Buea and the susceptibility of isolates to antibiotics. METHODS: One hundred and fifty clinical specimens (urine, wound and sputum) collected from patients attending various health institutions in Buea, and 50 environmental swabs from furniture, appliances and surroundings of these institutions were screened for P. aeruginosa using standard microbiological and biochemical methods. Antimicrobial susceptibility of the isolates was determined by the disc diffusion assay. RESULTS: Fifty-one (25.5%) of the 200 specimens were positive for the organism with urine (30%) being the most common source of isolation. Biochemical characterization grouped the isolates into eight biotypes with biotypes B III (25.59%), B II (23.53%) and B I (21.57%) being the most prevalent. Antimicrobial susceptibility results of isolates revealed 13 antibiotype patterns based on resistance to the antimicrobial agents investigated. The resistance pattern, cefotaxime, gentamicin and tetracycline (CTX(R) GEN(R) TET(R)) was the most common (21.6%) amongst the isolates. Over 40% of the isolates exhibited multi-drug resistance to five or more antibiotics, especially environmental isolates. However, there was a significant difference (P < 0.05) in the susceptibility of isolates to ciprofloxacin (98%), amikacin (90.2%) and netilmicin (80.4%) compared with other drugs used in the study. CONCLUSION: A combination of biotyping and antibiogram, which are relatively cheap and routinely available methods in our environment, could be useful for clinical and epidemiological studies particularly in laboratories in the developing world with limited resources.

Aged↗

Aminoglycoside resistance among Pseudomonas aeruginosa isolates with an unusual disk diffusion antibiogram.

In recent years, a number of clinical microbiology laboratories have isolated Pseudomonas aeruginosa with the unusual aminoglycoside disk diffusion result of resistance to both amikacin and gentamicin but susceptibility to tobramycin (ArGrTs). A total of 39 isolates of P. aeruginosa reported to have this resistance pattern were retested by the standard National Committee for Clinical Laboratory Standards disk diffusion procedure; 30 strains (77%) were confirmed to be ArGrTs. These 30 isolates were further examined for susceptibility to those aminoglycosides by agar dilution and broth micro- and macrodilution methods. Only 27, 27, and 23% of the isolates appeared to be ArGrTs by agar, broth microdilution, and broth macrodilution testing, respectively. Most of the remaining isolates were resistant to all three aminoglycosides when tested by broth dilution and resistant only to gentamicin when tested by agar dilution. The percentages of strains resistant to any particular aminoglycoside by agar dilution, broth microdilution, and broth macrodilution, respectively, were 43, 80, and 70 for amikacin, 97, 93, and 100 for gentamicin, 100, 100, and 100 for netilmicin, 30, 87, and 93 for sisomicin, and 13, 57, and 50 for tobramycin. These results indicate that strains showing the unusual aminoglycoside antibiogram are less susceptible to aminoglycosides in general and should probably be considered borderline resistant to all aminoglycosides. The efficacy of aminoglycosides in the treatment of infections produced by these strains is unknown.

Amikacin↗