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Revised disk diffusion interpretive criteria for cefaclor, loracarbef, cefprozil and cefixime when testing Haemophilus influenzae on haemophilus test medium.

The aim of the current five-center collaborative study was to reassess the interpretive criteria for cefaclor, loracarbef, cefprozil and cefixime previously adopted or proposed by the National Committee for Clinical Laboratory Standards (NCCLS) for disk diffusion susceptibility tests with Haemophilus influenzae on Haemophilus Test Medium (HTM) agar. MICs and zones of inhibition were determined using NCCLS methods, HTM and two collections of strains of Haemophilus influenzae. One group of strains consisted of 118 stock organisms taken largely from various recent U.S. antibiotic resistance surveillance studies. The emphasis in this selected group of organisms was on strains that were beta-lactamase negative but ampicillin resistant (BLNAR) by some other mechanism. The second collection of test organisms consisted of 50 recent clinical isolates of Haemophilus influenzae obtained from each of the five participating study centers. This group was considered representative of the type of Haemophilus influenzae currently recovered from clinical sources in the USA. Frequency distribution assessment and error-rate bounded analysis of scattergram comparisons of MICs and zone sizes were used to develop the following zone diameters interpretive for disk diffusion susceptibility tests with Haemophilus influenzae on HTM agar: cefaclor, > or = 20 mm (susceptible, S) and < or = 16 mm (resistant, R); loracarbef, > or = 19 mm (S) and < or = 15 mm (R); and cefprozil, > 18 mm (S) and < 14 mm (R). The respective MIC correlates for all three antimicrobial agents were < or = 8 micrograms/ml (S) and 32 micrograms/ml (R).(ABSTRACT TRUNCATED AT 250 WORDS)

Cefaclor↗

Disk diffusion susceptibility testing of Haemophilus influenzae using haemophilus test medium.

A three-center collaborative study was performed with the aim of determining the reproducibility of disk diffusion and broth microdilution susceptibility tests of Haemophilus influenzae with 19 different antimicrobial agents using a newly described medium, Haemophilus Test Medium. The results of both methods were found to be highly reproducible. Using Haemophilus Test Medium, inhibitory zone diameters and MICs were then determined for 150 test strains of Haemophilus influenzae, selected with an emphasis on known mechanisms of resistance. Regression analysis of zone diameter-MIC comparisons was used to define zone diameter interpretive criteria and MIC correlates for Haemophilus Test Medium disk diffusion tests performed with Haemophilus spp. for the following antimicrobial agents: ampicillin, amoxicillin/clavulanate, ampicillin/sulbactam, cefaclor, cefuroxime, cefamandole, ceftriaxone, cefotaxime, ceftizoxime, ceftazidime, cefixime, trimethoprim/sulfamethoxazole, rifampin, chloramphenicol, tetracycline, imipenem, aztreonam and ciprofloxacin.

Anti-Bacterial Agents↗

Isolation and characterization of a newly identified Haemophilus species from cats: "Haemophilus felis".

A gram-negative coccobacillus was isolated from the lower respiratory tract of a cat with chronic obstructive pulmonary disease. The isolate required CO2 and V factor for growth and was initially identified as Haemophilus paraphrophilus on the basis of its nutritional requirements, colony morphology, and some biochemical tests. Because of the host specificity of Haemophilus species and discrepancies in catalase, oxidase, and hemolytic activities, additional testing was done. Extensive biochemical testing, G+C content, and DNA reassociation studies indicated that the organism was distinct from other Haemophilus species. Therefore, the organism was identified as a previously unrecognized Haemophilus species and was tentatively named "Haemophilus felis." Bacteria identical to the original isolate were isolated from the nasopharynxes of 6 of 28 apparently normal cats, indicating that H. felis or H. felis-like organisms may be common members of the feline upper respiratory tract flora.

Animals↗

Response to immunization with Haemophilus influenzae type b polysaccharide-pertussis vaccine and risk of Haemophilus meningitis in children with the Km(1) immunoglobulin allotype.

In experimental animals, immune responses to certain antigens are regulated by immunoglobulin allotype-linked genes. In an effort to detect such genes in humans, we examined the antibody responses of 74 healthy children with different Km(1) or Gm(23) allotypes to a Haemophilus influenzae type b vaccine (type b polysaccharide capsule-pertussis vaccine). The anticapsular antibody responses of black or white children with the Km(1) allotype were 4.6- to 9.5-fold higher than those of children who lacked this determinant (P less than 0.004). No significant differences were found in antibody response with respect to the Gm(23) allotype. The frequencies of Km(1) and Gm(23) also were examined in 170 patients with Haemophilus meningitis, 71 patients with epiglottitis, and 173 control children. Km(1) was detected less frequently in black patients with meningitis (38%) than in those with epiglottitis (81%, P less than 0.002) or in controls (66%, P less than 0.0007). The relative risk of meningitis thus was 3.2-fold lower among black children with the Km(1) allotype than in those who lacked this allotype (odds ratio = 0.3, 95% confidence interval 0.2 to 0.6). However, the risk of meningitis was not decreased in white children with the Km(1) allotype (odds ratio = 1.0). There were no significant differences in the frequency of Gm(23) among the patient groups and controls. The Km(1) allotype but not the Gm(23) thus defines a subpopulation of children of both races who are high responders to this vaccine, and black children but not white children with the Km(1) allotype are at decreased risk of developing Haemophilus meningitis. These data indicate that in blacks, genes associated with Km(1) may affect immune response to a prototype type b Haemophilus vaccine, and perhaps interact with another factor related to race to affect susceptibility to Haemophilus meningitis.

Antibodies, Bacterial↗

DNA-based subtypes and antimicrobial susceptibility profiles of Haemophilus influenzae and Haemophilus parainfluenzae isolated from different tonsillar sites of children undergoing tonsillectomy and/or adenoidectomy.

We did a comparative analysis between DNA-based subtypes and antimicrobial susceptibility profiles on Haemophilus influenzae and Haemophilus parainfluenzae, isolated from multiple tonsillar sites per individual from patients with chronic recurrent tonsillitis and/or tonsillar idiopathic hypertrophy and undergoing tonsillectomy and/or adenoidectomy. A total of eighty-eight Haemophilus isolates were obtained aseptically from the surface and core of tonsils and/or adenoids of 32 out of 60 patients and identified at the species level by the X and V factors and the API NH Kit. The H. influenzae and H. parainfluenzae isolates as well as ATCC strains were tested for antimicrobial susceptibility using a panel of antimicrobial agents. Random amplified polymorphic DNA (RAPD) was done on extracted DNA from all Haemophilus isolates and ATCC strains, using one 10 mer and one 18 mer primers to subtype the two species. Antimicrobial susceptibility testing data have shown a variation in generated susceptibility patterns to tested antimicrobial agents among H. influenzae and H. parainfluenzae isolates. This variation was demonstrated too among isolates obtained from different tonsillar sites (core and surface) in a single patient. RAPD analysis identified 58/88 (66%) different RAPD patterns. Variations in RAPD patterns among H. influenzae and H. parainfluenzae were also observed in isolates obtained from different tonsillar sites of the same individual. A correlation between RAPD patterns and antimicrobial susceptibility data, have shown: 1) the predominance of one strain (RAPD pattern) of either Haemophilus species among isolated organisms per patient, and exhibiting different antimicrobial susceptibility profiles or 2) the existence of multiple strains (RAPD patterns) of either Haemophilis species per patient, and showing either a single or multiple antimicrobial susceptibility profile(s). These observations question the validity of swab cultures obtained from a single tonsillar site per patient, for detection, identification and determination of antimicrobial profiles of the etiology of tonsillitis, since swab specimens taken from only one site may or may not reflect the etiology of infection.

Adenoidectomy↗

[Neonatal infections caused by Haemophilus influenzae and Haemophilus parainfluenzae].

Haemophilus influenzae is a frequent causal agent of serious infections in infants. Mostly these infections occur at the age of 3 months to 3 years. Reports about infections in the neonatal period were rare in the past and most of them were single cases. In the last years an increase of neonatal infections due to Haemophilus influenzae had been reported. In the period from June 1980 to January 1986 we saw 5 newborns with severe infections due to Haemophilus influenzae and one neonate infected by Haemophilus parainfluenzae. By discussion of our cases and by review of the literature we want to show important aspects of pathogenesis, clinical symptoms, diagnostics and therapy of neonatal infections due to bacteria of the genus Haemophilus.

Haemophilus Infections↗

Haemophilus endocarditis: report of 42 cases in adults and review. Haemophilus Endocarditis Study Group.

To define the clinical, microbiological, and therapeutic characteristics of haemophilus endocarditis, we reviewed the charts of 42 adults with haemophilus endocarditis (native valve disease, 37; prosthetic valve disease, five) who were followed up between 1983 and 1995 in France. The mean duration of symptoms before diagnosis was 34 days. The causative Haemophilus species were as follows: H. parainfluenzae (26 adults), H. aphrophilus (9), H. paraphrophilus (4), and H. influenzae (3). According to the Duke criteria, 38 cases of endocarditis were definitive and four were possible. Thirty-nine patients received combination antibacterial therapy and three received therapy with a beta-lactam agent alone (mean duration, 46 days). Arterial embolism occurred in 15 patients. Cardiac surgery was indicated for 18 patients; 16 of these surgeries were performed within 3 months. Two patients died of heart failure. In conclusion, haemophilus endocarditis is rare and is mainly due to H. parainfluenzae. Although surgery is often necessary, haemophilus endocarditis has a favorable prognosis.

Adolescent↗

Antimicrobial susceptibility testing of less commonly isolated Haemophilus species using Haemophilus test medium.

Haemophilus test medium (HTM) was developed recently for dilution and disk diffusion antimicrobial agent susceptibility testing of Haemophilus influenzae. The application of HTM to the testing of other, less frequently encountered Haemophilus species recovered from humans was evaluated in this study by using commercially prepared HTM (BBL Microbiology Systems, Cockeysville, Md.) in broth microdilution and agar disk diffusion susceptibility tests with 18 antimicrobial agents. A total of 93.3% of 90 isolates belonging to six Haemophilus species provided acceptable growth in HTM agar disk diffusion tests, while only 63.3% (57 of 90) provided acceptable growth in the broth microdilution tests. However, HTM agar dilution testing provided an alternative method for those strains (primarily H. haemolyticus) which failed to grow adequately in broth. Based on the latest National Committee for Clinical Laboratory Standards guidelines (standard M2-T4) for interpretation of HTM disk tests of H. influenzae, the overall very major, major, and minor errors for all 18 drugs and six species tested were 0.2, 0.7, and 3.4%, respectively. Thus, the use of HTM in agar or broth susceptibility tests can be recommended for testing the less commonly encountered Haemophilus species by using the same test conditions and interpretive guidelines developed for H. influenzae.

Culture Media↗

Comparison of in-house and commercially prepared haemophilus test media for disk diffusion testing of ampicillin against Haemophilus species.

The performance characteristics of two lots of Haemophilus Test Medium (HTM) prepared in-house at Mayo Clinic, one lot obtained from BBL (Cockeysville, Md.) and one lot obtained from Remel (Lenexa, Kans.), for disk diffusion susceptibility testing of ampicillin against 81 Haemophilus strains were assessed. Insufficient growth occurred in 2.5 to 13.6% of strains, depending on the HTM used, and was most frequent for beta-lactamase-negative, ampicillin-resistant strains when they were tested with one lot of HTM prepared in-house. Results for all 14 beta-lactamase-positive Haemophilus strains were in complete agreement for all lots of HTM when either 1990 or 1993 National Committee for Clinical Laboratory Standards (NCCLS) interpretive guidelines were followed. However, these beta-lactamase-positive Haemophilus strains could be as precisely, but more rapidly, identified by the cefinase disk method. Results for 56 beta-lactamase-negative, ampicillin-susceptible strains and 11 beta-lactamase-negative, ampicillin-resistant strains varied significantly among all HTM lots tested and as to whether 1990 or 1993 NCCLS interpretive guidelines were followed. We conclude that in our hands disk diffusion testing with HTM prepared in-house or obtained from two commercial sources and following either 1990 or 1993 NCCLS guidelines is an unacceptable method for determining the susceptibilities of Haemophilus strains to ampicillin.

Ampicillin↗

Genetic and phenotypic comparison of three new avian Haemophilus-like taxa and of Haemophilus paragallinarum Biberstein and White 1969 with other members of the family Pasteurellaceae Pohl 1981.

In the course of post-mortem bacteriological examinations, several previously unreported bacterial strains were isolated from budgerigars, pigeons, kestrels, and a goose. They have been separated into three distinct collectives according to their cultural, morphological, and biochemical characteristics. Since they require V factors, they were tentatively assigned to the genus Haemophilus Winslow et al. 1917. This preliminary classification was checked by determination of guanine + cytosine contents and genome sizes and by DNA:DNA hybridization tests among reference strains of the three new avian taxa and recognized species of the family Pasteurellaceae Pohl 1981. With the same methods, the genetic relationships of Haemophilus paragallinarum Biberstein and White 1969 within the family were determined. It could be shown that the three avian Haemophilus-like taxa have to be regarded as new species within the family Pasteurellaceae not affiliated with the recognized genera Actinobacillus, Haemophilus and Pasteurella. H. paragallinarum must be excluded from the genus Haemophilus because of its closer relationship to the actinobacilli. All strains investigated can be differentiated from each other and from recognized species of Pasteurellaceae using an appropriate set of biochemical tests.

Animals↗

EFFECT OF INTERSPECIFIC TRANSFORMATION ON LINKAGE RELATIONSHIPS OF MARKERS IN HAEMOPHILUS INFLUENZAE AND HAEMOPHILUS PARAINFLUENZAE.

Nickel, Lois (University of Pennsylvania, Philadelphia), and Sol H. Goodgal. Effect of interspecific transformation on linkage relationships of markers in Haemophilus influenzae and Haemophilus parainfluenzae. J. Bacteriol. 88:1538-1544. 1964.-Highly competent cells of Haemophilus parainfluenzae were obtained by a modification of the aerobic-"anaerobic" method of Goodgal and Herriott. The mutant markers for streptomycin and novobiocin resistance in H. parainfluenzae were shown to be unlinked. These same markers were transferred into H. influenzae in two transformation steps, and the deoxyribonucleic acid (DNA) was extracted. Transformation of recipient H. influenzae with DNA from the latter showed the markers to be linked, and transformation of recipient H. parainfluenzae with this DNA showed that the markers were not cotransferred. The linked streptomycin- and novobiocin-resistance mutations of H. influenzae were transformed into H. parainfluenzae in two stages, and the DNA was extracted. Transformation of recipient H. parainfluenzae with this DNA demonstrated the markers to be unlinked in H. parainfluenzae. When these markers, originally from H. influenzae and present as unlinked factors in H. parainfluenzae, are transferred back into H. influenzae, the markers showed their original linkage. The novobiocin- and erythromycin-resistance markers of H. influenzae were unlinked in H. influenzae, but were linked when transferred into H. parainfluenzae. The erythromycin-resistance marker of H. influenzae, when transferred into a novobiocin-resistant mutant of H. parainfluenzae, showed linkage with it. Evidence is presented to show that H. parainfluenzae streptomycin and novobiocin markers are homologous to streptomycin and novobiocin markers in H. influenzae.

DNA↗

Molecular characterization of chloramphenicol-resistant Haemophilus parainfluenzae and Haemophilus ducreyi.

We examined chloramphenicol-resistant Haemophilus parainfluenzae and Haemophilus ducreyi strains isolated in various parts of the world. The antibiotic resistance determinants were located on conjugative plasmids in H. ducreyi, but were chromosomally located in H. parainfluenzae. Both species produced chloramphenicol acetyltransferases (CATs) that were sensitive to 5,5'-dithiobis(2-nitrobenzoic acid) like the enteric type II and Haemophilus influenzae CAT enzymes, but differed from these enzymes in elution patterns and subunit molecular weight. Southern blot analysis showed the H. parainfluenzae and H. ducreyi CAT genes were molecularly related to the enteric type II class as well as the H. influenzae CAT. Heterogeneity of the physiochemical properties of the CATs was observed; however, the data suggested that all three Haemophilus spp. have a common ancestral source for the CATs.

Acetyltransferases↗

Introduction of transposon Tn916 DNA into Haemophilus influenzae and Haemophilus parainfluenzae.

Enterococcus (Streptococcus) faecalis transposon Tn916 was introduced into Haemophilus influenzae Rd and Haemophilus parainfluenzae by transformation and demonstrated to transpose efficiently. Haemophilus transformants resistant to tetracycline were observed at a frequency of approximately 3 x 10(2) to 5 x 10(3)/micrograms of either pAM120 (pGL101::Tn916) or pAM180 (pAM81::Tn916) plasmid DNAs, which are incapable of autonomous replication in this host. Restriction enzyme analysis and Southern blot hybridization revealed that (i) Tn916 integrates into many different sites in the H. influenzae and H. parainfluenzae genomes; (ii) only the 16.4-kilobase-pair Tn916 DNA integrates, and no vector DNA was detected; and (iii) the Tetr phenotype was stable in the absence of selective pressure. Second-generation Tn916 transformants occurred at the high frequency of chromosomal markers and retained their original chromosomal locations. Similar results were obtained with H. influenzae Rd BC200 rec-1 as the recipient strain, which suggests host rec functions are not required in Tn916 integrative transposition. Transposition with Tn916 is an important procedure for mutagenesis of Haemophilus species.

DNA Transposable Elements↗

Evaluation of two commercial kits and arbitrarily primed PCR for identification and differentiation of Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus, and Haemophilus paraphrophilus.

The closely related species Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus, and Haemophilus paraphrophilus are common findings in oral microbiota. The aims of this study were to evaluate the applicability of the Rapid NH and API ZYM kits and arbitrarily primed PCR (AP-PCR) in the identification and differentiation of the three species from each other. The material included 62 clinical isolates and three reference strains of A. actinomycetemcomitans representing the 5 serotypes and 18 AP-PCR genotypes. Haemophilus species included 12 clinical isolates and 11 reference strains of H. aphrophilus, H. paraphrophilus, and 5 other species. For the PCR amplification, the oligonucleotide 5'-CAGCACCCAC-3' was used as a primer. Contrary to the consistent performance of API ZYM, the Rapid NH system was able to identify only 10 of 65 (15%) A. actinomycetemcomitans isolates, whereas all Haemophilus species were correctly identified. The API ZYM test differentiated A. actinomycetemcomitans from H. aphrophilus and H. paraphrophilus by negative beta-galactosidase and alpha-glucosidase reactions and a positive esterase lipase reaction. However, the API ZYM test was unable to differentiate H. aphrophilus from H. paraphrophilus, it also could not differentiate A. actinomycetemcomitans serotypes from each other. Among the H. aphrophilus isolates three AP-PCR genotypes and among H. paraphrophilus isolates only one AP-PCR genotype, distinct from those of A. actinomycetemcomitans, were found. The Rapid NH test showed poor ability to identify clinical isolates of all A. actinomycetemcomitans serotypes. Moreover, AP-PCR genotyping proved to be a rapid method for the species differentiation of A. actinomycetemcomitans, H. aphrophilus, and H. paraphrophilus.

Actinobacillus Infections↗

In vitro activity of a new broad-spectrum, beta-lactamase-stable oral cephalosporin, cefixime, in comparison with other drugs, against Haemophilus influenzae, Haemophilus parainfluenzae, Moraxella catarrhalis and Streptococcus pneumoniae.

Cefixime, a new orally absorbed iminomethoxyaminothiazolyl cephalosporin, was tested against some microorganisms involved in upper and lower respiratory tract infections such as Haemophilus (influenzae and parainfluenzae), Moraxella catarrhalis and Streptococcus pneumoniae, isolated in the period from November 1990 to April 1991. Its activity was compared to nine other antimicrobial agents: erythromycin, trimethoprim-sulfamethoxazole, ampicillin, amoxicillin-clavulanate, cefaclor, ceftazidime, ceftriaxone, cefotaxime and ofloxacin. Cefixime inhibits 90% of S. pneumoniae, H. influenzae and H. parainfluenzae, both beta-lactamase producers (BLP) or not (NBLP) at concentrations of less than 0.25 mg/l. It inhibits 90% of M. catarrhalis (BLP and NBLP) at concentrations of less than 1 mg/l. In general, cefixime has a superior in vitro activity with respect to cefaclor and the other cephalosporins as well as erythromycin and amoxicillin (the last one in BLP strains). In the evaluation of the antibacterial activity of beta-lactam against Haemophilus and M. catarrhalis, the authors observed different indications in the guidelines for ampicillin. Cefixime is not destroyed by the plasmid-mediated beta-lactamase produced by Haemophilus sp. and M. catarrhalis (TEM and ROB in Haemophilus strains and BRO in M. catarrhalis). In view of its excellent in vitro activity against the commonly encountered respiratory tract pathogens, cefixime is indicated in the therapy of these infections.

Administration, Oral↗

Comparison of Wilkins-Chalgren medium supplemented with nicotinamide adenine dinucleotide and Haemophilus test medium for susceptibility testing of clinical Haemophilus isolates.

Haemophilus test medium (HTM) was compared with Wilkins-Chalgren agar (WCA; supplemented with 15 micrograms/ml nicotinamide adenine dinucleotide (NAD)) for antibiotic susceptibility testing of 74 clinical isolates of Haemophilus influenzae and Haemophilus parainfluenzae. The Bauer-Kirby agar disk diffusion method and the agar dilution procedure were the two tests employed. WCA + NAD and HTM yielded comparable results for the following antimicrobial drugs and drug combinations: ampicillin, amoxicillin plus clavulanic acid, ampicillin plus sulbactam, cefotaxime, cefuroxime, chloramphenicol, ciprofloxacin, and doxycycline. However, WCA + NAD slightly antagonized rifampin. Co-trimoxazole was significantly antagonized by WCA + NAD. WCA + NAD was much simpler with regard to in-house preparation, and WCA plates could be supplemented with NAD following pouring, solidification and storage of the plates simply by surface spreading the required amount of NAD with a glass spatula, thus adding considerably to laboratory flexibility. It is suggested that additional laboratories comparatively and critically examine WCA + NAD medium for antibiotic susceptibility testing of clinical Haemophilus isolates.

Culture Media↗

Invasive serotype a Haemophilus influenzae infections with a virulence genotype resembling Haemophilus influenzae type b: emerging pathogen in the vaccine era?

OBJECTIVE: Haemophilus influenzae type b causes severe disease in nonimmune infants and young children; other serotypes are uncommon pathogens and thought to have low virulence. Some have hypothesized that with the virtual elimination of H influenzae type b, other serotypes might acquire virulence traits and emerge as important pathogens of children. We describe the clinical, epidemiologic, and molecular biologic features of 5 cases of severe disease attributable to Haemophilus influenzae type a. METHODS: After observing 4 cases of invasive disease caused by H influenzae type a, we reviewed microbiology records at 3 reference laboratories that perform all serotyping in Utah and surveillance databases. Strains of H influenzae type a and control strains were examined by Southern blotting with the use of the cap probe pUO38 and by pulsed-field gel electrophoresis. The putative virulence mutation, the IS1016-bexA deletion, was detected by polymerase chain reaction amplification and sequencing. RESULTS: During a 10-month period, we observed 5 children with severe invasive disease caused by H influenzae type a. No isolates of H influenzae type a had been submitted to the reference laboratories between 1992 and 1998. The median age of patients was 12 months (range: 6-48 months). Four of 5 had meningitis and bacteremia; 1 had purpura fulminans. Three isolates, representing 1 of 2 pulsed-field gel electrophoresis patterns, contained the IS1016-bexA deletion and were associated with particularly severe disease. CONCLUSIONS: We describe an unusual cluster of severe disease caused by H influenzae type a that resembles the clinical and epidemiologic features of H influenzae type b disease. Our data support the hypothesis that the IS1016-bexA deletion may identify more virulent strains of H influenzae. Haemophilus influenzae, epidemiology, virulence, serotyping, pathogenicity.

Base Sequence↗

Escherichia coli ATCC 35218 as a quality control isolate for susceptibility testing of Haemophilus influenzae with haemophilus test medium.

Current National Committee for Clinical Laboratory Standards (NCCLS) susceptibility guidelines for quality control testing with Haemophilus influenzae do not include a beta-lactamase-producing strain that could detect the deterioration of the beta-lactamase inhibitor components of amoxicillin-clavulanic acid, ampicillin-sulbactam, and piperacillin-tazobactam. The objective of the study was to determine if comparable quality control results for Escherichia coli ATCC 35218, a beta-lactamase-producing strain, would be produced for the three beta-lactam-beta-lactamase inhibitor agents with Haemophilus test medium and Mueller-Hinton medium. The criteria used in this study to determine if Haemophilus test medium was acceptable for quality control testing of E. coli ATCC 35218 was that 100% of the results obtained with an antimicrobial agent-methodology combination needed to be within the acceptable NCCLS ranges established with Mueller-Hinton medium. The MIC testing results obtained by the broth microdilution and E-test methods with amoxicillin-clavulanic acid and piperacillin-tazobactam were all within the NCCLS ranges; however, the results obtained with ampicillin-sulbactam by both methods were not within the NCCLS ranges. Acceptable results were obtained by the disk diffusion methodology with ampicillin-sulbactam and piperacillin-tazobactam but not with amoxicillin-clavulanic acid. When performing susceptibility testing with H. influenzae with the beta-lactam-beta-lactamase inhibitors, in addition to quality control testing with H. influenzae ATCC 49247, testing of E. coli ATCC 35218 on Haemophilus test medium is an effective way to monitor the beta-lactamase inhibitors in some antimicrobial agent-methodology combinations.

Culture Media↗