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In Vitro Susceptibility of Nocardia Spp. to a New ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, Dec. 1989, p. 2140-2141 Vol. 33, No. 12 0066-4804/89/122140-02$02.00/0 Copyright © 1989, American Society for Microbiology In Vitro Susceptibility of Nocardia spp. to a New Fluoroquinolone, Tosufloxacin (T-3262) KATSUKIYO YAZAWA, YUZURU MIKAMI,* AND JUN UNO Department ofExperimental Chemotherapy, Research Center for Pathogenic Fungi and Microbial Toxicoses, Chiba University, 1-8-1, Inohana, Chiba 280, Japan Received 31 July 1989/Accepted 11 September 1989 The in vitro activity of a new fluoroquinolone, tosufloxacin (T-3262), against 111 pathogenic Nocardia strains was studied by an agar dilution method and compared with the activities of other fluoroquinolones. AU strains were susceptible to tosufloxacin, and the drug was 2 to 20 times more active than four other quinolones tested. Nocardia farcinica was the Nocardia species most susceptible to tosufloxacin. Nocardia spp. are generally regarded as opportunistic ology (8). A third species, N. nova, was proposed by pathogens, especially in immunocompromised hosts. The Tsukamura (11). Therefore, in this study, all clinical isolates reported frequency of infection with Nocardia spp. is in- and stocked cultures of the N. asteroides group were divided creasing in Japan and the United States (1, 10, 14). into three species, i.e., N. asteroides sensu stricto (17 Although sulfonamides remain the agents of first choice strains), N. farcinica (31 strains), and N. nova (15 strains). against Nocardia species, the effectiveness of minocycline The group was divided on the basis of such tests as raffinose and amikacin has been reported (3, 11). The potential and sodium citrate utilization, susceptibility to 5-fluoroura- usefulness of other antimicrobial agents such as herbekacin cil, and growth at 42°C (6, 8, 11). N. brasiliensis (33 strains), (14), imipenem, and combinations of cephems and ,B-lacta- N. otitidis-caviarum (15 strains), and type strains of each mase inhibitors has also been determined by in vitro suscep- species were also included in the present comparisons. tibility tests (5). Nocardia spp. grown in Sabouraud dextrose agar (Difco The fluoroquinolones are currently being evaluated in Laboratories, Detroit, Mich.) slants were inoculated into a clinical studies and appear to have a wide spectrum of 10-ml Erlenmeyer shake flask containing 5 ml of 3% glucose- activity against a variety of bacterial infections (13). The in enriched brain heart infusion broth (Difco) and eight 4-mm vitro activity of some fluoroquinolones against Nocardia glass beads intended to reduce aggregation of the nocardial spp. has recently been reported (2, 4). Tosufloxacin (T-3262; cells. The cultures were Fig. 1) is a fluoroquinolone recently developed in Japan. It is grown for 72 h on a rotary shaker characterized by having the 2,4-difluorophenyl and 3-amino- (5.8-cm stroke) at 27°C. They were then diluted with sterile 1-pyrrolidinyl groups in its molecule. It has been found to be saline to give 108 CFU. The plates were spotted with a clinically useful for various infections, such those of the multipoint inoculator A 400 (Denley Instruments Ltd., Sus- respiratory tract (7, 9). sex, England) that delivered 0.005 ml, which resulted in a Our study was designed to assess the in vitro activity of spot inoculum of approximately 2 x 105 CFU. The inocu- tosufloxacin against Nocardia spp. (including the N. aster- lated plates were incubated at 27°C for 7 days and then were oides group [N. asteroides sensu stricto, N. farcinica, and examined for growth to determine the MIC. N. nova], N. brasiliensis, and N. otitidis-caviarum) and The activities of the quinolones tested against five Nocar- compare it with those of four other fluoroquinolones. dia species are shown in Table 1. In general, all of them The quinolones used in this study were tosufloxacin showed activity against all Nocardia species. However, the (Toyama Chemical Co. Ltd., Kanazawa, Japan), norfloxacin MICs of each drug varied greatly depending on the species. (Torii Seiyaku, Tokyo, Japan), ofloxacin (Daiichi Seiyaku, N. farcinica was the most susceptible, followed by N. Tokyo, Japan), and ciprofloxacin (Bayer Yakuhin, Osaka, brasiliensis and N. otitidis-caviarum. N. asteroides sensu Japan). Enoxacin was chemically synthesized. Purified to- stricto and N. nova were resistant. sufloxacin was dissolved in 0.1 N NaOH and diluted with On a weight basis, tosufloxacin was the most active sterilized saline. Other drugs were dissolved in dimethyl quinolone, with an MIC for 90% of N. farcinica strains of sulfoxide and then diluted with saline. The MIC, defined as 0.57 ,ug/ml. Ciprofloxacin and ofloxacin were slightly less the active than tosufloxacin. Present data regarding the MICs of lowest drug concentration resulting in complete inhibi- ciprofloxacin, ofloxacin, and enoxacin are in general agree- tion of visible growth, was determined by an agar dilution ment with those of other investigators (2, 5); therefore, it method using Sensitivity-Disk agar (Eiken Chemical Co. was confirmed that tosufloxacin is the most active fluoroqui- Ltd, Tokyo, Japan) to give final concentrations ranging from nolone against N. asteroides reported to date. Tsukamura et 0.2 to 100 ,ug/ml for all quinolones. Drug-free plates were al. (12) reported the therapeutic effect of ofloxacin on included as a bacterial growth control. tuberculosis: when was to A total of 111 Nocardia strains were used. The species N. pulmonary the drug given 19 asteroides has patients with chronic cavitary lung tuberculosis in a single been considered heterogeneous in its physio- daily dose of 300 mg for 6 to 8 months, a decrease in the logical and biochemical character and was actually divided number of tubercle bacilli in the sputum was observed in into N. asteroides sensu stricto and N. farcinica in the almost all individuals, and negative conversion occurred in recently published Bergey's Manual of Systematic Bacteri- five people. To our knowledge, no clinical trials using fluoroquinolones against Nocardia infections have been re- * Corresponding author. ported. However, since the clinical features of Nocardia 2140 VOL. 33, 1989 NOTES 2141 also indicate that imipenem is active only against the N. asteroides group and not against N. brasiliensis or N. F4 otitidis-caviarum. This species-specific activity of imipenem may be due to the Nocardia species-specific inactivation N N N H3C ) S03H.H20 enzyme P-lactamase. Therefore, most of the problems re- ported to date of variable susceptibility of the N. asteroides group to different drugs appear to be resolved the division COOH by 0 of the N. asteroides group into three species. Since no enzymatic inactivation of fluoroquinolones has yet been FIG. 1. Structure of tosufloxacin (tosufloxacin tosilate). reported, these variabilities may be due to the difference in permeation ability of the drugs in each species. Studies on the ability of tosufioxacin to permeate various Nocardia infections, especially those caused by the N. asteroides species are now in progress in our laboratory with labeled group, are similar to those of Mycobacterium tuberculosis tosufloxacin. infection, tosufloxacin may serve as a clinically effective alternative agent for patients suffering from N. farcinica LITERATURE CITED infection who are unable to tolerate therapy with sulfon- 1. Beaman, B. L., J. Burnside, B. Edward, and W. Causey. 1976. Nocardial infections in the United States, 1972-1974. J. Infect. amides. Dis. 134:286-289. We recently reported that deoxystreptamine-containing 2. Berkey, P., D. Moore, and K. Rolston. 1988. In vitro suscepti- aminoglycoside antibiotics such as gentamicin, kanamycin, bilities of Nocardia species to newer antimicrobial agents. and tobramycin show variable activities depending on the Antimicrob. Agents Chemother. 32:1078-1079. species of Nocardia they are used against (14). We also 3. Cockerill, R. F., R. S. Edson, G. D. Roberts, and J. C. Waldorf. suggested that these variabilities are due largely to the 1984. Trimethoprim-sulfamethoxazole-resistant Nocardia aster- inactivation enzymes, such as aminoglycoside acetyltrans- oides causing multiple hepatic abscesses. Am. J. Med. 77: ferase [AAC(3)], in N. farcinica (14). Our preliminary data 558-560. 4. Gombert, M. E., T. M. Aulicino, L. DuBouchet, and L. R. Berkowitz. 1987. Susceptibility of Nocardia asteroides to new quinolones and P-lactams. Antimicrob. Agents Chemother. 31: TABLE 1. Antinocardial activity of tosufloxacin and other 2013-2014. quinolones against 111 strains of pathogenic Nocardia spp. 5. Gombert, M. E., T. M. Aulicino, L. DuBouchet, G. E. Silver- man, and W. M. Sheinbaum. 1986. Therapy of experimental Organism Drug MIC (Wg/ml)a cerebral nocardiosis with imipenem, amikacin, trimethoprim- (n) Range 70% 90%O sulfamethoxazole, and minocycline. Antimicrob. Agents Che- mother. 30:270-273. N. asteroides (17) Enoxacin 0.78->100 30.2 83.3 6. Goodfellow, M., and D. E. Minnikin. 1981. The genera of Norfloxacin 3.13->100 >100 >100 Nocardia and Rhodococcus, p. 2016-2027. In M. P. Starr, H. Ofloxacin 0.39->100 11.0 >100 Stolp, H. G. Trupper, A. Balows, and H. J. Schlegel (ed.), The Ciprofloxacin <0.2-100 12.1 56.7 prokaryotes, vol. 2. Springer-Verlag KG, Berlin. Tosufloxacin <0.2-50 7.5 16.4 7. Katahira, J., T. Kumada, K. Totsuka, and K. Shimizu. 1988. Basic and clinical studies on T-3262. Chemotherapy (Tokyo) N. farcinica (31) Enoxacin 1.56-12.5 5.7 9.7 36(Suppl. 9):418-423. (In Japanese.) Norfloxacin 3.13-25 13.3 21.1 8. Lechevalier, H. A. 1986. Nocardioforms, p. 1458-1506. In Ofloxacin 0.39-3.13 1.56 2.6 P. H. A. Sneath (ed.), Bergey's manual of systematic bacteri- Ciprofloxacin <0.2-3.13 0.96 1.4 ology, vol. 2. The Williams & Wilkins Co., Baltimore. Tosufloxacin <0.2-3.13 <0.39 0.57 9. Nasu, M., T. Yamazaki, H. Yamazaki, Y. Kuroda, Y. Goto, H. Shigeno, J. Goto, T. Tashiro, H. Miyako, S. Hiro-oka, K.
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