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Evaluation of clonal relatedness of extended-spectrum beta-lactamase-producing Proteus mirabilis isolates by quantitative antibiogram and RAPD typing.

OBJECTIVE: To delineate, using two different typing systems, the clonal relatedness of 40 isolates of extended-spectrum beta-lactamase (ESbetaIa)-producing Proteus mirabilis obtained over a period of 7 years in six hospitals in the Paris area and two in Pas-de-Calais. METHODS: Random amplified polymorphic DNA (RAPD) polymerase chain reaction typing was applied by using three random primers on the ESbetaIa-producing P. mirabilis isolates and on isogenic Escherichia coli strains with or without plasmids encoding the representative resistance pattern transferred from P. mirabilis. Quantitative antibiogram typing, which was also applied to the P. mirabilis isolates, was used to define the euclidean distance between these strains. RESULTS: After having demonstrated that P. mirabilis plasmids did not influence chromosomal DNA amplification, we could classify the ESbetaIa-producing P. mirabilis isolates into 12 groups based on RAPD fingerprints. The same isolates were classified into 19 groups by quantitative antibiogram typing. Despite this difference in group numbers, general concordance between the typing systems was observed. This allowed us to show that the greater number of isolates in some hospitals belonged to a single strain and that single isolates obtained in different hospitals generally represented unique strains. CONCLUSIONS: A small number of ESbetaIa-producing P. mirabilis strains was isolated during 7 years in the eight medical centers studied, and the number of different strains identified suggested that inter-hospital transfer had not occurred.

Journal Article↗

Molecular analysis by pulsed-field gel electrophoresis and antibiogram of Streptococcus pneumoniae serotype 6B isolates from selected areas within the United States.

Fifty-eight clinical isolates of Streptococcus pneumoniae serotype 6B, including 16 from Alaska, 14 from Arizona, 11 from Washington, and 17 from seven additional states, were analyzed. The antibiograms of these isolates were assigned to 10 antibiotic profiles based on their susceptibilities to penicillin, erythromycin, tetracycline, and trimethoprim-sulfamethoxazole. Thirty-two (55%) of these isolates were penicillin nonsusceptible, while 21 (36%) were intermediate or resistant to three or more antibiotics. The restriction endonucleases ApaI and SmaI were used to digest intact chromosomes, and the fragments were resolved by pulsed-field gel electrophoresis (PFGE). The ApaI and SmaI PFGE patterns were combined, and 13 of the 16 Alaskan isolates showed indistinguishable PFGE patterns. One other isolate exhibited highly related ApaI and SmaI PFGE patterns, differing by only one band after restriction with ApaI. Among the 14 isolates from Arizona, 1 was indistinguishable from the predominant ApaI and SmaI PFGE patterns seen in the Alaskan isolates; 5 others were highly related (+/-1 band after cutting with either enzyme) to the Alaskan isolates, suggesting a common ancestral origin. Of the remaining eight isolates, six additional ApaI plus SmaI PFGE patterns were observed. The 28 isolates from the various contiguous states had 22 ApaI plus SmaI PFGE patterns. No correlations were found between specific PFGE patterns, antibiograms, dates of isolation, or geography. The serotype 6B isolates across the contiguous United States were genetically diverse, while the 6B isolates from Alaska appeared to be much less diverse.

Alaska↗

Extensive hospital-wide spread of a multidrug-resistant enterobacter cloacae clone, with late detection due to a variable antibiogram and frequent patient transfer.

A hospital-wide increase in the number of patients with aminoglycoside-resistant Enterobacter cloacae (AREC) isolated from clinical cultures was detected in December 2002 using a classical surveillance system (CSS). CSS refers to a strategy based on the recognition of an increased incidence of a species with a particular antibiogram at certain wards in a limited period. Since clonal spread was suspected, hospital records were reviewed for E. cloacae culture-positive patients. Based upon genotyping of 139 clinical E. cloacae isolates from 80 patients, it was concluded that 53 patients had had clinical cultures with a single AREC clone since April 2001. Determinants for unnoticed spread were investigated retrospectively, as was the possibility that a computer-assisted surveillance method would have detected this outbreak at an earlier stage. Determinants associated with late detection of clonal spread were the following: (i) the absence of a hospital-wide increase in incidence of E. cloacae cases for 1.5 years, (ii) the long time interval between cases, (iii) the hospital-wide occurrence of new cases, due to a high number of patient transfers between wards, (iv) the large variety of clinical sites, and (v) the high variability of antibiograms (n = 33). Retrospective application of a recently described computer-assisted surveillance method as well as an "in-house"-developed algorithm resulted in earlier detection of the outbreak of 6 and 12 months, respectively. These findings suggest that computerized tools for surveillance may recognize resistance trends that are too complex to be detected by manual review and indicate the need for prospective evaluation of such algorithms.

Algorithms↗

Identification of multiresistant Staphylococcus epidermidis in neonates of a secondary care hospital using pulsed field gel electrophoresis and quantitative antibiogram typing.

AIMS: To determine the diversity of types of Staphylococcus epidermidis in a neonatal care unit of a secondary care hospital in the Netherlands. METHODS: In a prospective study, specimens from nose, ear, axilla, umbilicus, and groin were taken from patients twice a week during a period of up to two weeks. All isolates were typed by both pulsed field gel electrophoresis (PFGE) and antibiogram analysis. RESULTS: Fifty three S epidermidis isolates from 15 of 24 patients were obtained in one to four surveys. Fourteen isolates from six patients had a common PFGE pattern and were of one multiresistant antibiogram type. The remaining 39 isolates were allocated to 24 sporadic PFGE types and were more susceptible to antibiotics. Colonisation with the multiresistant strain correlated with a long period of stay and with the use of specific antibiotics. The multiresistant isolates were related closely to isolates of S epidermidis found in a recent study in a teaching hospital in the vicinity of the secondary care hospital. CONCLUSION: Repeated sampling and the use of two typing methods allowed the identification of two closely related multiresistant S epidermidis strains in two hospitals in the same area.

Bacterial Typing Techniques↗

Using hospital antibiogram data to assess regional pneumococcal resistance to antibiotics.

Antimicrobial resistance to penicillin and macrolides in Streptococcus pneumoniae has increased in the United States over the past decade. Considerable geographic variation in susceptibility necessitates regional resistance tracking. Traditional active surveillance is labor intensive and costly. We collected antibiogram reports from North Carolina hospitals and assessed pneumococcal susceptibility to multiple agents from 1996 through 2000. Susceptibility in North Carolina was consistently lower than the national average. Aggregating antibiogram data is a feasible and timely method of monitoring regional susceptibility patterns and may also prove beneficial in measuring the effects of interventions to decrease antimicrobial resistance.

Anti-Bacterial Agents↗

[Bacterial infections in cancer patients: what should we treat them with when the result of antibiogram is not yet known?].

Cancer patients are prone to bacterial infections, which frequently on account of severity require prompt administration of antibiotics--even before the result of antibiogram is known. 500 antibiograms obtained from cultures taken from 307 cancer patients treated in General Hospital no. 2, Bielsko-Biala during 1991-1995 were analyzed. Marked diversity in isolated bacterial strains was found, the dominant pathogens being Gram positive species (mainly Staphylococci and Streptococci, which constituted 46.2% of all isolated strains). The most frequent Gram negative species were Escherichia coli, Proteus spp., Klebsiella spp. and Enterobacter spp. Analysis of antibiotic efficiency suggests that cancer patients in case of infection should be treated empirically as follows: 1. Moderate infection--monotherapy with broad spectrum antibiotic, covering also Gram positive bacteria--such as ciprofloxacin . 2. Severe infection--combination therapy consisting of two antibiotics: one with activity against Gram positive bacteria such as cloxacillin, erythromycin, cefazolin, cephalexin, clindamycin--the other with activity against Gram negative bacteria, such as aminoglycoside (e.g., netilmicin, amikacin) or third generation cephalosporine (e.g., cefotaxime, ceftriaxone).

Anti-Bacterial Agents↗

Incidence of post operative wound infection and their antibiogram in a teaching and referral hospital.

A total of 406 post-operative clean wounds were studied for the presence of sepsis and antibiogram of organisms were established. The over all post-operative sepsis rate was 13% (clinical) and 12% (bacteriological). Staphylococcus aureus (32%) and Pseudomonas species (21%) were the commonest organisms recovered and Netilmycin, Cephaloridine and Norfloxacin were the most effective antibiotics against both Gram positive and negative infections. This study reflects the change in pattern of infecting bacterial flora in case of post operative wound infections and its antibiogram.

Adult↗

Antibiogram of Klebsiella pneumoniae isolates from Buea, Cameroon.

OBJECTIVE: To determine the antibiotic susceptibility of K. pneumoniae isolates from Buea, Cameroon. DESIGN: A prospective study of K. pneumoniae isolates from clinical samples of nosocomial origin. SETTING: A laboratory based investigative study at the Biotechnology Centres of the Universities of Buea and Yaounde 1, Cameroon, and three Buea based hospitals. K. pneumoniae isolates were obtained from sputum, wound swabs and urine and screened for their antibiogram using standard procedures. RESULTS: Results on the antibiogram showed seven distinct antibiotypes distinguished by different susceptibilities to aminoglycosides (Spectinomycin and Gentamicin), Chloramphenicol and Augmentin. All the isolates shared multi-resistance to Amoxicillin and Trimethoprim. However, the isolates showed marked susceptibilities to Norfloxacin (90.01%), Cefuroxime (95.45%) and Ciprofloxacin (86.36%). CONCLUSION: The study has revealed that K. pneumoniae isolates in the environment of Buea, Cameroon are multi-drug resistant. This finding is of clinical and epidemiological significance.

Cameroon↗

Antibiogram and beta-lactamase production of Staphylococcus aureus isolates from different human clinical specimens in Edo State, Nigeria.

The antibiogram and Beta-lactamase of 73 isolates of Staphylococcus aureus from 235 different human clinical specimens were determined using standard procedures. These various clinical specimens were got from teaching hospitals and some private hospitals in Edo State. The results of the antibiogram showed 100% susceptibiity to Vancomycin, 78.1% to Gentamicin, 71.3% to Chloramphenicol, 69.8% to Erythromycin and 61.6% to Cloxacillin. The results of the beta-lactamase detection showed that 84.1% of the isolates were penicillinase positive, which probably accounted for the 100% resistance obtained for both Ampicillin and Penicillin. This thus suggests that clinicians should enlighten patients on the consequences of indiscriminate use of Penicillin and other antimicrobial agents.

Anti-Bacterial Agents↗

Characterisation, biotyping, antibiogram and klebocin typing of Klebsiella with special reference to Klebsiella oxytoca.

400 strains of Klebsiellae identified by culture characteristics and biochemical reactions were subjected to biotyping, antibiogram and klebocin typing. Based on indole production, pectin and gelatin liquefaction 16.0% of all the isolates were Klebsiella oxytoca. Maximum sensitivity was shown to Amikacin (72%) and maximum resistance to Ampicillin (87.5%). Klebocin typability was 73.5%. So by combining biotyping, antibiogram and Klebocin typing, Klebsiella could be differentiated better than based on any single marker.

Anti-Bacterial Agents↗

Isolation, phage typing and antibiogram of Salmonella from man and animals in northeastern India.

BACKGROUND AND OBJECTIVE: Salmonella is an important zoonotic pathogen and its prevalence in the animals acts as a continuous threat to man. The present study was carried out to report the isolation along with the serotypes, phage types and antibiogram pattern of Salmonella among man, livestock and poultry in the northeastern India. METHODS: A total of 654 samples from diarrhoeic livestock and humans were processed for the isolation of Salmonella. All the isolates were subjected to antibiogram studies against 15 antimicrobials. Representative isolates of S. Typhimurium and S. Enteritidis were phage typed. RESULTS: Ninety five isolates of Salmonella enterica belonging to 5 serotypes- S. Typhimurium, S. Enteritidis, S. Gallinarum, S. Paratyphi B and S. Bareilly were obtained with an overall prevalence rate of 14.40 per cent. S. Typhimurium isolates were distributed among four phages- DT003, DT004, DT096 and DT193 and all the S. Enteritidis isolates belonged to a single phage type, PT13a/7. Interspecies sharing of the phages was observed. Norfloxacin, enrofloxacin, gentamycin and ciprofloxacin were most effective, whereas, doxycycline, ampicillin, amoxycillin and tetracycline were relatively less effective. INTERPRETATION AND CONCLUSION: Our findings showed that three of the five serovars as well as some of the phage types of these serovars were shared by animals and humans indicating the zoonotic potential of the organism. Thus, it is imperative that salmonellosis control measures adopted for humans should give adequate importance to its control in the animals particularly their products.

Animals↗

A statistical method to evaluate the reliability and reproducibility of a standardised technique for the conduct of antibiograms on two species of Enterobacteriaceae.

Antibiograms are only as reliable as is permitted by the quality of the specimen, the reliability or reproducibility of the method used and the ability of the clinician to interpret the results. Non-standardised methods may give erroneous results. Some of these procedural effects were investigated by means of statistical analysis of variance to determine the effect on the reliability of the test. To ensure reliable and reproducible test results with antibiograms, it is imperative that the type of sensitivity test medium as well as the density of the inoculum of the test organisms be standardised.

Analysis of Variance↗

[Salmonella montevideo from a hospital outbreak with different antibiograms determined by a plasmid].

From May to June 1984 an outbreak of Salmonella montevideo occurred in the neonatal ward of a district's hospital. 26 babies were involved and mild forms of disease were observed in the most cases. The infants were hospitalized from different reasons and treated chemotherapeutically prior to the nosocomial infection. The Salmonella montevideo strains isolated from the faeces of the infants showed five different antibiograms. In all strains the multiple drug resistance was determined by a single plasmid belonging to the incompatibility group C and with a molecular weight of 110 Md or 100 Md in the threefold resistant strains. These plasmids were found also in the faecal flora (E. coli and Klebsiella) of all patients. It is suggested that a transfer of the plasmids from E. coli or Klebsiella to Salmonella montevideo occurred under in vivo condition, since distinct patients excreted susceptible as well as resistant Salmonellae. This example shows the necessity to include antibiogram and plasmid analysis as epidemiological markers.

Cross Infection↗

Comparative study of the biotype, hemolysin-producing capability and antibiogram of the aquatic and the clinical strains of Aeromonas hydrophila.

One hundred and seventy-two Aeromonas hydrophila strains were isolated from aquatic hosts including fish, turtle, frogs, shellfish and water itself. The aquatic organisms were compared with 45 clinical strains with regard to biotype, hemolysin, and antibiogram. The carrier rate of A. hydrophila in the aquatic environment was 61.7%. Of the aquatic strains 51% were hemolysin producers, equivalent to a capability for producing enterotoxin. The hemolysin-producing capability of aquatic strains was not significantly different from that of the clinical strains. The main code numbers or biotypes of aquatic strains were 3265 (29.4%), 3065 (17.5%), 7065 (9.6%), 7265, 3225 (each 8.5%). The main code number or biotypes of clinical strains were 3265 (36.1%), 3065 (9.8%), 7261 (6.6%), 2265, 3261 and 6241 (each 4.9%). Analysis of the antibiotic susceptibility pattern of both aquatic and clinical A. hydrophila strains using the microbroth dilution method showed that there is no significant difference in antibiotic susceptibility. Most aquatic and clinical strains were highly susceptible to tobramycin, gentamicin, amikacin, cefuroxime, piperacillin, cefotaxime and cefoperazone. Most strains of aquatic A. hydrophila were highly susceptible to cefamandole and chloramphenicol; however, clinical strains were only moderately susceptible to the same drugs. Most aquatic and clinical strains were highly resistant to ampicillin, and moderately resistant to sulbenicillin. Clinical strains were moderately resistant to cephapirin; however, aquatic strains were moderately susceptible to cephapirin. Based on the above findings, we may considered aquatic environment be, at least partially, the source of clinical strains. Furthermore, biotypes of aquatic A. hydrophila do not correspond to antibiogram or hemolysin-producing patterns.

Aeromonas↗

[Nature and antibiotic resistance of bacteria encountered in an intensive care unit. Computerized study based on 422 antibiograms].

A computer analysis of 422 sheets of data from antibiograms performed on 146 patients in a resuscitation unit of multipurpose type, was performed. The study comprises the investigation of the nature of the isolated bacteria and the resistance of the organisms to antibiotics alone or in combinations. The results obtained show on the one hand, that there is a development over time of resistance to antibiotics and a change in the nature of the bacteria, and on the other, that a precise knowledge of the bacteriological data concerning a given department enables the intiation of treatment before having the antibiogram results with an increased chance of efficacy. The importance of such a study is controversial.

Anti-Bacterial Agents↗

[Computer management of antibiogram data (author's transl)].

Numerous models of computer management of antibiogram data are reviewed with particular regard to their clinical significance. A system presently used in the institute of the A. is taken into particular consideration. The most characteristic aspect of this system is the possibility of retaining via computer memory all the data obtained without having to reveal the information to the attending physician. This model allows the establishment and maintenance of a complete and constant exchange of information with the clinician who can utilize a large part of the data obtained from the antibiogram.

Computers↗

[Evaluation of a new system for simultaneous identification and antibiograms of enterobacteria from urine].

A new rotor for the ABAC system has been investigated that allows the main enterobacteria of urinary source to be identified (this is performed manually) and simultaneously the relative antibiogram (this is automatically carried out). The precision of this identification has been shown using control strains previously identified by three miniaturized kits: API 20 E, MICRO-ID and ENTEROPLATE. The accuracy of the ten identification tests present in the new "Identibiogramma" rotor has also been shown. Finally, the correspondence has been shown between the antibiogram performed with the automatized method and that with the KirbyBauer method. The data obtained evidence the validity of the new rotor and of the ABAC system.

Anti-Bacterial Agents↗

[Considerations to antibacterial chemo-therapy antecedent to the isolation of the causative agent and evaluation of the antibiogram (author's transl)].

The principles are discussed, which should serve as a basis for therapeutical decisions antecedent to the isolation of the causative agent and evaluation of the antibiogram. These concern the degree of urgency for chemotherapy, the frequency of the association of bacterial species with defined clinical pictures, the actual distribution of antibiotic resistance, the situation of the patient's immune defence system, and side effects of the drug. Possibilities of a "calculated" therapy in urinary tract infections, bronchitis, enteritis, septicaemia, and post-operative wound infection are explained. Considerations about the frequency of causative agents in certain bacterial infections and figures about the actual resistance distribution within bacteria species commonly connected with the disease in quesiton form the baseline for the choice of a chemotherapeutic until the antibiogram is performed.

Anti-Bacterial Agents↗