ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, Apr. 1985, p. 646-647 Vol. 27, No. 4 0066-4804/85/040646-02$02.00/0 Copyright X 1985, American Society for Microbiology In Vitro Susceptibilities and 1-Lactamase Production of 53 Clinical Isolates of Branhamella catarrhalis SALVADOR ALVAREZ,',2* MARY JONES,2 SHIRLEY HOLTSCLAW-BERK,2 JUAN GUARDERAS,2 AND STEVEN L. J3ERK",2 Infectious Disease Section, Department of Medicine, Veterans Administration Medical Center,1 and the Quillen-Dishner College of Medicine,2 Johnson City, Tennessee 37684 Received 31 October 1984/Accepted 22 January 1985 We tested 53 clinical isolates of Branhamella catarrhalis recovered from patients with respiratory symptoms to determine the susceptibility of the isolates to 25 antimicrobial agents, including the newer I8-lactam antibiotics. Of the 53 strains, 46 (86.7%) were P-lactamase producers. All the strains were susceptible to the majority of the new penicillins and cephalosporins.- The combinations of amoxacillin-clavulanic acid and ticarcillin-clavulanic acid were also very active against the P-lactamase-prodiicing strains. Branhamella catarrhalis, formerly known as Neisseria the microbroth dilution technique with Mueller-Hinton broth catarrhalis, is generally considered a harmless oropharyn- supplemented with cations to a final concentration of 25 mg geal commensal bacterium. However, in the past few years of Mg2+ per liter and 50 mg of Ca2+ per liter (11). The it has been recognized as the etiologic agent of otitis media in preparation and inoculation of the microbroth plates was children (9) and has been isolated in pure culture from accomplished with the MIC 2000 system (Dynatech Labora- patients with acute bronchitis and pneumonia (2, 7, 10, 12, tories, Inc., Alexandria, Va.), using an inoculum of 105 16, 17), acute laryngitis (15), acute sinusitis (1), septicemia CFU/ml. The MIC was defined as the lowest concentration (3), meningitis (3, 13), and endocarditis (6, 14). of the drug which yielded no visible growth after 18 to 24 h P-Lactamase production by B. catarrhalis has occurred in of incubation at 35°C. 4 to 100% of isolates (5, 7-9, 16). Despite this fact, very little A total of 46 strains (86.7%) of B. catarrhalis produced information is available on the susceptibility of B. ca- 1-lactamase as determined by the chromogenic cephalospo- tarrhalis to antimicrobial agents, especially the newer 1B- rin disk. lactam antibiotics. The results of testing the susceptibility of the isolates to 25 We report here on the antimicrobial susceptibility and the drugs are summarized in Table 1. Non-,B-lactamase- incidence of 1-lactamase in 53 clinical isolates of B. ca- producing strains were inhibited by low amounts of penicil- tarrhalis from hospitalized patients with respiratory disease. lin G, but ,B-lactamase producers were uniformly resistant to Organisms. A total of 53 clinical isolates of B. catarrhalis penicillin G. Some strains with the ability to produce 13- were obtained from hospitalized patients with lower respira- lactamase were inhibited by low amounts of ampicillin. tory symptoms admitted to the Veterans Administration Repeated testing of these strains with a larger inoculum Medical Center, Johnson City, Tenn. The isolates were yielded similar results. obtained from blood, transtracheal aspirates, and expecto- Nearly all the strains of B. catarrhalis were resistant to rated sputum from patients with pneumonia or acute exac- penicillin G, methicillin, amoxicillin, and vancomycin. All 53 erbation of chronic bronchitis. The clinical isolates were strains were susceptible to ticarcillin, piperacillin, azlocillin, identified by conventional methods previously described (4): cefoxitin, cefuroxime, cefotiam, cefbuperazone, moxalac- gram-negative diplococci, oxidase production, growth on tam, cefoperazone, ceftazidime, and cefotaxime. On a weight 5% sheep blood agar incubated at 37°C under humidified basis, cefbuperazone, ceftazidime, moxalactam, piperacil- 10% C02, lack of pigmentation, failure to produce acid from lin, azlocillin, amoxicillin-clavulanic acid, and ticarcillin- glucose, maltose, and sucrose, and reduction of nitrate to clavulanic acid were the most active drugs among the nitrite. ,B-lactam antibiotics. P-Lactamase production. The production of ,B-lactamase Erythromycin, tetracycline, chloramphenicol, and trimeth- was determined in each strain by the use of the chromogenic oprim-sulfamethoxazole were uniformly active against the cephalosporin disk nitrocefin (Cefinase; BBL Microbiology B. catarrhalis isolates. Systems, Cockeysville, Md.). Several investigators have reported that clinically signifi- Antibiotics. Antibiotics were kindly provided by the man- cant isolates of B. catarrhalis produce ,B-lactamase (5, 7-9, ufacturers as reagent-grade powders and included penicillin 16). In our studying, 46 of 53 strains of B. catarrhalis G, ampicillin, methicillin, cephalothin, cefamandole, cefoxi- isolated from hospitalized patients with lower respiratory tin, cefuroxime, cefotiam, moxalactam, cefoperazone, cefo- symptoms were found to produce 1-lactamase. The 13- taxime, ceftazidime, cefbuperazone, ticarcillin, piperacillin, lactamase of B. catarrhalis is not plasmid mediated but azlocillin, erythromycin, chloramphenicol, tetracycline, van- rather of chromosomal origin and intracellular (5). comycin, trimethoprim-sulfamethoxazole, amoxicillin-potas- was the least active antibiotic tested. sium clavulanic acid, and ticarcillin-potassium clavulanic Penicillin Ampicillin, acid. cephalothin, and cefamandole had a similar susceptibility Antibiotic testing. MICs were determined pattern. ,B-lactamase production was not an absolute predic- susceptibility by tor of susceptibility because $ome of the 1-lactamase- producing strains were susceptible to ampicillin. The new * Corresponding author. penicillins and the second- and third-generation cephalospo- 646 NOTES 647 VOL. 27, 1985 CITED TABLE 1. Antibactrial activity of 25 antibiotics against 53 strains LITERATURE of B. catarrhalis MIC (,ug/ml) 1. Brorson, J. E., A. Axelsson, and S. E. Holms. 1976. Studies on with special Antibiotic Range 50% 90% Branhamella catarrhalis (Neisseria catarrhalis) reference to maxillary sinusitis. Scand. J. Infect. Dis. 8:151-155. 2.0 8.0 2. Diamond, L. A., and B. Lorber. 1984. Branhamella catarrhalis Penicillin G <0.125-16.0 associa- Ampicillin <0.125-4.0 0.25 1.0 pneumonia and immunoglobulin abnormalities: a new Amoxicillin <0.25-8.0 2.0 8.0 tion. Am. Rev. Respir. Dis. 129:876-878. 2.0 16.0 3. Doern, G. V., M. J. Miller, and R. E. Winn. 1981. Branhamella Methicillin <0.25-16.0 re- Ticarcillin <0.5-16.0 1.0 8.0 (Neisseria) catarrhalis systemic disease in humans. Case Piperacillin <0.125-0.25 <0.125 0.125 ports and review of the literature. Arch. Intern. Med. Aziocillin <0.125-0.25 <0.125 <0.125 141:1690-1692. <0.06 0.25 4. Doeen, G. V., and S. A. Morse. 1980. Branhamella (Neisseria) Amoxicillin plus <0.06-0.25 Mi- Clavulanic acid catarrhalis: criteria for laboratory identification. J. Clin. Ticarcillin plus <0.05-0.5 <0.5 <0.5 crobiol. 11:193-195. Clavulanic acid 5. Doern, G. V., K. G. Siebers, I. M. Hallick, and S. A. Morse. Clindamycin 0.125-2.0 1.0 1.0 1980. Antibiotic susceptibility of beta-lactamase-producing Erythromycin <0.06-0.25 0.125 0.25 strains of Branhamella (Neisseria) catarrhalis. Antimicrob. Vancomycin <0.5-64 16.0 32.0 Agents Chemother. 17:24-29. Cephalothin <0.5-8.0 4.0 4.0 6. Douer, D., Y. Danziger, and J. Pinkhas. 1977. Neisseria ca- 2.0 4.0 tarrhalis endocarditis. Ann. Intern. Med. 86:116. Cefamandole 0.25-4.0 Branha- Cefoxitin <0.06-0.25 0.125 0.125 7. Johnson, M. A., W. L. Drew, and M. Roberts. 1981. Cefuroxime 0.125-1.0 0.5 1.0 mella (Neisseria) catarrhalis-a lower respiratory tract patho- Cefotiam 0.125-1.0 1.0 1.0 gen? J. Clin. Microbiol. 13:1066-1069. Cefbuperazone <0.125-0.125 <0.125 <0.125 8. Kallings, I., S. Bengtsson, P. Christensen, S. E. Holm, L. Lind, Moxalactam <0.125-0.125 <0.125 <0.125 and M. Kalin. 1983. Antibiotic sensitivity of Haemophilus Cefoperazone <0.125-2.0 0.25 1.0 influenzae, Streptococcus pneumoniae, Streptococcus pyo- Cefotaxime <0.125-0.5 0.25 0.5 genes, and Branhamella catarrhalis isolated from upper respi- Ceftazidime <0.06-0.125 <0.06 <0.06 ratory tract infections in Sweden. Scand. J. Infect. Dis. Suppl. Trimethoprim- <0.25-4.75-0.25-4.75 <0.25-4.75 <0.25-4.75 39:100-105. sulfamnethoxazole 9. Kovatch, A. L., E. R. Wald, and R. H. Michaels. 1983. Beta- Tetracycline <0.06-0.25 0.125 0.125 lactamase-producing Branhamella catarrhalis causing otitis me- Chloramphenicol 0.25-0.50 0.50 0.50 dia in children. J. Pediatr. 102:261-264. 10. Louis, M. H., E. L. Gabay, G. E. Mathisen, and S. M. Finegold. 1983. Branhamella catarrhalis pneumonia. West. J. Med. 138:47-49. 11. National Committee for Clinical Laboratory Standards. 1983. M7-T. National Committee for Clinical Laboratory Standards, rins were the most active drugs in vitro against B. catarrh- tetracy- Villanova, Pa. alis. Other groups of drugs such as erythromycin, 12. Ninane, G., J. Joly, and M. Kraytman. 1978. Bronchopulmonary cline, and trimethoprim-sulfamethoxazole were also active. infection due to Branhamella catarrhalis: 11 cases assessed by The combination of clavulanic acid with ticarcillin or amoxi- transtracheal puncture. Br. Med. J. 1:276-278. cillin was very effective in vitro. 13. Pfister, I. E., M. V. Gallager, T. G. Potterfield, and D. W. Based on the results of the MIC determinations of 25 Brown. 1965. Neisseria catarrhalis bacteremia with meningitis. for 53 clinically significant isolates of B. ca- J. Am. Med. Assoc. 193:399-401. antibiotics 1976. Neisseria catarrhalis and the increase in the percentage of ,B-lactamase- 14. Pollock, A. A., and R. S. Holzman. tarrhalis Ann. Intern. Med. 85:206-207. strains, it appears best to avoid the use of either endocarditis. producing 15. Schalen, L., P. Christensen, C. Kamme, H. Mioerner, K. Pett- penicillin or ampicillin in the treatment of disease caused by of Branhamella broad- ersson, and C.