Chapter 31 : Antimicrobial Susceptibility Testing of Bacterial Agents of Bioterrorism: Strategies and Considerations

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, species, , , , and are bacterial species that can produce deadly infections; collectively they have caused epic plagues and human misery for centuries . This chapter explores the progress made over the last three years to establish both standardized susceptibility testing methods and interpretive criteria for ,, , , , and species. Although a standardized testing method has been published by CLSI for , two issues remain controversial. The first is whether a 0.5 McFarland inoculum contains an adequate number of cells for testing, since colony counts performed from a standardized inoculum are often below 5 × 105CFU/ml, which is the standard inoculum size for broth microdiution assays. The second issue focuses on the role of the chromosomally encoded β-lactamases produced by in mediating penicillin resistance, and whether the β-lactamases are constitutively produced or inducible in most anthrax strains. Antimicrobial susceptibility testing methods established by Clinical and Laboratory Standards Institute (CLSI) for MIC testing of follow those used for other ; however, in studies conducted by Centers for Disease Control and Prevention (CDC), adequate growth of some isolates was not achieved in Mueller-Hinton broth in 24 hrs. Antimicrobial susceptibility testing methods used for studies vary widely in the literature. Antimicrobial susceptibility testing of bacterial agents of bioterrorism provides critical data for guiding prophylaxis or treatment of patients exposed to or infected with pathogens.

Citation: Tenover F. 2005. Antimicrobial Susceptibility Testing of Bacterial Agents of Bioterrorism: Strategies and Considerations, p 414-422. In White D, Alekshun M, McDermott P (ed), Frontiers in Antimicrobial Resistance. ASM Press, Washington, DC. doi: 10.1128/9781555817572.ch31

Key Concept Ranking

Antimicrobial Susceptibility Testing
Disk Diffusion Test
Gram-Negative Bacilli
Francisella tularensis
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1. Athamna, A.,, M. Athamna,, N. Abu-Rashed,, B. Madlej,, D. J. Bast,, and E. Rubenstein. 2004. Selection of Bacillus anthracis isolates resistant to antibiotics J. Antimicrob. Chemother. 54:424428.
2. Baker, C. N.,, D. G. Hollis,, and C. Thornsberry. 1985. Antimicrobial susceptibility testing of Francisella tularensis with a modified Mueller-Hinton broth. J. Clin. Microbiol. 22:212215.
3. Bosch, J.,, J. Linares,, M. J. Lopez de Goicoechea,, J. Ariza,, M. C. Cisnal,, and R. Martin. 1986. In vitro activity of ciprofloxacin, ceftriaxone, and five other antimicrobial agents against 95 strains of Brucella melitensis. J. Antimicrob. Chemother. 17: 459461.
4. Bodur, H.,, N. Balaban,, S. Aksaray,, V. Yetener,, E. Akinci,, A. Colpan,, and A. Erbay. 2003. Biotypes and antimicrobial susceptibilities of Brucella isolates. Scand. J. Infect. Dis. 35:337338.
5. Boulanger, L. L.,, P. Ettestad,, J. D. Fogarty,, D. T. Dennis,, D. Romig,, and G. Mertz. 2004. Gentamicin and tetracyclines for the treatment of human plague: review of 75 cases in New Mexico, 1985-1999. Clin. Infect. Dis. 38:663669.
6. Brown, S. D.,, K. Krisher,, and M. M. Traczewski. 2004. Broth microdilution susceptibility testing of Francisella tularensis: quality control limits for nine antimicrobial agents and three standard quality control strains. J. Clin. Microbiol. 42:58775880.
7. Centers for Disease Control and Prevention. 2001. Update: Investigation of bioterrorism-related anthrax and interim guidelines for exposure management and antimicrobial therapy, October 2001. Morb. Mortal. Wkly. Rep. 50:909918.
8. Chen, Y.,, J. Succi,, F. C. Tenover,, and T. Koehler. 2003. β-lactamase genes of the penicillin-susceptible Bacillus anthracis Sterne strain. J. Bacteriol. 185:823830.
9. Choe, C. H.,, S. S. Bouhaouala,, I. Brook,, T. B. Elliot,, and J. Knudson. 2000. In vitro development of resistance to ofloxacin and doxycycline in Bacillus anthracis Sterne. Antimicrob. Agents Chemother. 44:1766.
10. Clinical and Laboratory Standards Institute. 2005. Performance Standards for Antimicrobial Susceptibility Testing. Fifteenth Informational Supplement M100-S15. Clinical and Laboratory Standards Institute, Wayne, Pa.
11. Coker, P. R.,, K. L. Smith,, and M. E. Hugh-Jones. 2002. Antimicrobial susceptibilities of diverse Bacillus anthracis isolates. Antimicrob. Agents Chemother. 46:38433845.
12. Dennis, D. T.,, and J. M. Hughes. 1997. Multidrug resistance in plague. N. Engl. J. Med. 337:702704.
13. Dennis, D. T.,, T. V. Inglesby,, D. A. Henderson,, J. G. Bartlett,, M. A. Ascher,, E. Eitzen,, A. D. Fine,, A. M. Friedlander,, J. Hauer,, M. Layton,, S. R. Lillibridge,, J. E. McDade,, M. T. Osterholm,, T. O’Toole,, G. Parker,, T. M. Perl,, P. K. Russell,, K. Tonat, for the Working Group on Civilian Biodefense. 2001. Tularemia as a biological weapon. JAMA. 285:27632773.
14. Dixon, T. C.,, M. Meselson,, J. Guillemin,, and P. C. Hanna. 1999. Anthrax. N. Engl. J. Med. 341:815826.
15. Doganay, M.,, and N. Aydin. 1991. Antimicrobial susceptibility of Bacillus anthracis. Scand. J. Infect. Dis. 23:333335.
16. Drago, L.,, E. De Vecchi,, A. Lombardi,, L. Nicola,, M. Valli,, and M. R. Gismondo. 2002. Bactericidal activity of levofloxacin, gatifloxacin, penicillin, meropenem, and rokitamycin against Bacillus anthracis clinical isolates. J. Antimicrob. Chemother. 50: 10591063.
17. Eickhoff, T. C.,, J. V. Bennett,, P. S. Hayes,, and J. Feeley. 1970. Pseudomonas pseudomallei susceptibility to chemotherapeutic agents. J. Infect. Dis. 121:95102.
18. Ellis, J.,, P. C. F. Oyston,, M. Green,, and R. W. Titball. 2002. Tularemia. Clin. Microbiol. Rev. 15:631646.
19. Evans, A. S.,, and P. S. Brachman (ed.). 1998. Bacterial Infections of Humans: Epidemiology and Control. Plenum Publishing Corporation, New York, N.Y.
20. Friedlander, A. M.,, S. L. Welkos,, M. L. M. Pitt,, J. W. Ezzell,, P. L. Worsham,, K. J. Rose,, B. E. Ivins,, J. R. Lowe,, G. B. Howe,, P. Mikesell,, and W. B. Lawrence. 1993. Postexposure prophylaxis against experimental inhalation anthrax. J. Infect. Dis. 167: 12391242.
21. Galimand, M.,, A. Guiyoule,, G. Gerbaud,, B. Rasoamanana,, S. Chanteau,, E. Carniel,, and P. Courvalin. 1997. Multidrug resistance in Yersinia pestis mediated by a transmissible plasmid. N. Engl. J. Med. 337:677680.
22. Garcia del Blanco, N.,, C. B. Gutiérrez Martin,, V. A. de la Puenta Redondo,, and E. F. Rodríguez Ferri. 2004. In vitro susceptibility of field isolates of Francisella tularensis subsp. holarctica recovered in Spain to several antimicrobial agents. Res. Vet. Sci. 76:195198.
23. Garcia-Rodriguez, J. A.,, J. L. Monoz Bellido,, M. J. Fresnadillo,, and I. Trujillano. 1993. In vitro activities of new macrolides and rifapentine against Brucella spp. Antimicrob. Agents Chemother. 37:911913.
24. Gilbert, D. N.,, R. C. Moellering, Jr.,, G. M. Eliopoulos,, and M. A Sande. 2004. The Sandford Guide to Antimicrobial Therapy—2004. Antimicrobial Therapy, Inc., Hyde Park, Vt.
25. Godfrey, A. J.,, S. Wong,, D. A. B. Dance,, W. Chaowagul,, and L. E. Bryan. 1991. Pseudomonas pseudomallei resistance to β-lactam antibiotics due to alterations in the chromosomally encoded β-lactamase. Antimicrob. Agents Chemother. 35:16351640.
26. Gotuzzo, E.,, and C. Carrillo,. 2004. Brucella, p. 17171724. In S. L. Gorbach,, J. G. Bartlett,, and N. R. Blacklow (ed.), Infectious Diseases, 3rd ed. Lippincott Williams, & Wilkins, Philadelphia, Pa.
27. Guiyoule, A.,, B. Rasoamanana,, C. Buchrieser,, P. Michel,, S. Chanteau,, and E. Carniel. 1997. Recent emergence of new variants of Yersinia pestis in Madagascar. J. Clin. Microbiol. 35: 28262833.
28. Heine, H. S.,, M. J. Englan,, D. M. Wang,, and W. R. Byrne. 2001. In vitro antibiotic susceptibilities of Burkholderia mallei (causative agent of glanders) determined by broth microdilution and Etest. Antimicrob. Agents Chemother. 45:21192121.
29. Ikäheimo, I.,, H. Syrjälä,, J. Karhukorpi,, R. Schildt,, and M. Koskela. 2000. In vitro antibiotic susceptibility of Francisella tularensis isolated from humans and animals. J. Antimicrob. Chemother. 46:287290.
30. Inglesby, T. V.,, D. T. Dennis,, D. A. Henderson,, J. G. Bartlett,, M. S. Ascher,, E. Eitzen,, A. D. Fine,, A. M. Friedlander,, J. Hauer,, J. F. Koerner,, M. Layton,, J. McDade,, M. T. Osterholm,, T. O’Toole,, G. Parker,, T. M. Perl,, P. K. Russell,, M. Schoch-Spana,, and K. Tonat. 2000. Plague as a biological weapon: medical and public health management. JAMA. 283:22812290.
31. Inglesby, T. V.,, D. A. Henderson,, J. G. Bartlett,, M. S. Ascher,, E. Eitzen,, A. M. Friedlander,, J. Hauer,, J. McDade,, M. T. Osterholm,, T. O’Toole,, G. Parker,, T. M. Perl,, P. K. Russell,, and K. Tonat. 1999. Anthrax as a biological weapon: medical and public health management. JAMA. 281:17351745.
32. Ivans, B. E.,, S. I. Welkos,, G. B. Knudson,, and D. J. LeBlanc. 1988. Transposon Tn916 mutagenesis in Bacillus anthracis. Infect. Immun. 56:176181.
33. Jenny, A. W.,, G. Lum,, D. A. Fisher,, and B. J. Currie. 2001. Antibiotic susceptibility of Burkholderia pseudomallei from tropical northern Australia and implications for therapy of melioidosis. Int. J. Antimicrob. Agents 17:109113.
34. Jernigan, J. A.,, D. S. Stephens,, D. A. Ashford,, C. Omenaca,, M. S. Topiel,, M. Galbraith,, M. Tapper,, T. L. Fisk,, S. Zaki,, T. Popovic,, R. F. Meyer,, C. P. Quinn,, S. A. Harper,, S. K. Fridkin,, J. J. Sejvar,, C. W. Shepard,, M. McConnell,, J. Guarner,, W. J. Shieh,, J. M. Malecki,, J. L. Gerberding,, J. M. Hughes,, B. A. Perkins, and the Anthrax Bioterrorism Investigation Team. 2001. Bioterrorism-related inhalational anthrax: the first 10 cases reported in the United States. Emerg. Infect. Dis. 7:933944.
35.Johansson A., L. Beglund, L. Gothefors, A. Sjostedt, and A. Tarnvik. 2000. Ciprofloxacin for treatment of tularemia in children. Pediatr. Infect. Dis. J. 19:449453.
36. Kenny, D. J.,, P. Russell,, D. Rogers,, S. M. Eley,, and R. W. Titball. 1999. In vitro susceptibilities of Burkholderia mallei in comparison to those of other pathogenic Burkholderia spp. Antimicrob. Agents Chemother. 43:27732775.
37. Khan, M.Y.,, M. Dizon,, and F. W. Kiel. 1989. Comparative in vitro activities of ofloxacin, difloxacin, ciprofloxacin, and other selected antimicrobial agents against Brucella melitensis. Antimicrob. Agents Chemother. 33:14091410.
38.Kim, H-S., E-C. Choi, and B-K. Kim. 1993. A macrolidelincosamide- streptogramin B resistance determinant from Bacillus anthracis 590: cloning and expression of ermJ. J. Gen. Microbiol. 139:601607.
39. Kocagoz, S.,, M. Akova,, B. Altun,, D. Gur,, and G. Hascelik. 2001. In vitro activities of new quinolones against Brucella melitensis isolated in a tertiary care hospital in Turkey. Clin. Microbiol. Infect. 8:240242.
40. Klietman, W. F.,, and K. L. Ruoff. 2001. Bioterrorism: implications for the clinical microbiologist. Clin. Microbiol. Rev. 14: 364381.
41. Lalitha, M. K.,, and M. K. Thomas. 1997. Penicillin resistance in Bacillus anthracis. Lancet 349:1522.
42. Landinez, R.,, J. Linares,, E. Loza,, J. Martinez-Beltran,, and F. Baquero. 1992. In vitro activity of azithromycin and tetracycline against 358 clinical isolates of Brucella melitensis. Eur. J. Clin. Microbiol. 11:265267.
43. Lightfoot, N. F.,, R. J. D. Scott,, and P. C. B. Turnbull. 1990. Antimicrobial susceptibility of Bacillus anthracis. Salisbury Med. Bull. 68(Suppl.):9598.
44. Limaye, A. P.,, and C. J. Hooper. 1999. Treatment of tularemia with fluoroquinolones: two cases and a review. Clin. Infect. Dis. 29:922924.
45. Lopez-Merino, A.,, A. Contreras-Rodriguez,, R. Migranas-Ortiz,, R. Orrantia-Gradin,, G. M. Hernandez-Oliva,, A. T. Gutierrez- Rubio,, and O. Cardenosa. 2004. Susceptibility of Mexican brucella isolates to moxifloxacin, ciprofloxacin and other antimicrobials used in the treatment of human brucellosis. Scand. J. Infect. Dis. 36:636638.
46. Materon, I. C.,, A. M. Queenan,, T. M. Koehler,, K. Bush,, and T. Palzkill. 2003. Biochemical characterization of β-lactamase Bla1 and Bla2 from Bacillus anthracis. Antimicrob. Agents Chemother. 47:20402042.
47. Mohammed, M. J.,, C. K. Marston,, T. Popovic,, R. S. Weyant,, and F. C. Tenover. 2002. Antimicrobial susceptibility testing of Bacillus anthracis: comparison of results obtained by using the National Committee for Clinical Laboratory Standards broth microdilution reference and Etest agar gradient diffusion methods. J. Clin. Microbiol. 40:19021907.
48. Moore, R. A.,, D. DeShazer,, S. Reckseidler,, A. Wissman,, and D. E. Woods. 1999. Efflux-mediated aminoglycoside and macrolide resistance in Burkholderia pseudomallei. Antimicrob. Agents Chemother. 43:465470.
49. Meyerhoff, A.,, R. Albrecht,, J. M. Meyer,, P. Dionne,, K. Higgins,, and D. Murphy. 2004. US Food and Drug Administration approval of ciprofloxacin hydrochloride for management of postexposure inhalational anthrax. Clin. Infect. Dis. 39:303308.
50. National Committee for Clinical Laboratory Standards. 2003. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically. 6th ed., vol. 20, no. 2. Approved Standard M7-A6. National Committee for Clinical Laboratory Standards, Wayne, Pa.
51. National Committee for Clinical Laboratory Standards. 2001. Performance Standards for Antimicrobial Susceptibility Testing. Eleventh Informational Supplement M100-S11. National Committee for Clinical Laboratory Standards, Wayne, Pa.
52. National Committee for Clinical Laboratory Standards. 2004. Performance Standards for Antimicrobial Susceptibility Testing. Fourteenth Informational Supplement M100-S14. National Committee for Clinical Laboratory Standards, Wayne, Pa.
53. Pappas, G.,, S. Seitaridis,, N. Akritidis,, and E. Tsianos. 2004. Treatment of brucella spondylitis: lessons from an impossible meta-analysis and initial report of efficacy of a fluoroquinolone- containing regimen. Int. J. Antimicrob. Agents. 24:502507.
54. Patra G.,, J. Vaissaire,, M. Weber-Levy,, C. Le Doujet,, and M. Mock. 1998. Molecular characterization of Bacillus strains involved in outbreaks of anthrax in France in 1997. J. Clin. Microbiol. 36:34123414.
55. Perez-Castrillon, J. L.,, P. Bachiller-Luque,, M. Martin-Luquero,, F. J. Mena-Martin,, and V. Herreros. 2001. Tularemia epidemic in Northwestern Spain; clinical description and therapeutic response. Clin. Infect. Dis. 33:573576.
56. Piliouras, P.,, G. C. Ulett,, C. Ashhurst-Smith,, R. G. Hirst,, and R. E. Norton. 2002. A comparison of antibiotic susceptibility testing methods for cotrimoxazole with Burkholderia pseudomallei. Int. J. Antimicrob. Agents. 19:427429.
57. Price, L. B.,, A. Volger,, T. Pearson,, and P. Keim. 2003. In vitro selection and characterization of Bacillus anthracis mutants with high-level resistance to ciprofloxacin. Antimicrob. Agents Chemother. 47:23622365.
58. Rubinstein, E.,, R. Lang,, B. Shasha,, B. Hagar,, L. Diamanstein,, G. Joseph,, M. Anderson,, and K. Harrison. 1991. In vitro susceptibility testing of Brucella melitensis to antibiotics. Antimicrob. Agents Chemother. 35:19251927.
59. Ruhfel, R. E.,, N. J. Robillard,, and C. B. Thorne. 1984. Interspecies transduction of plasmids among Bacillus anthracis, B. cereus, and B. thuringiensis. J. Bacteriol. 157:708711.
60. Smith, M. D.,, D. X. Vinh,, T. T. Nguyen,, J. Wain,, D. Thung,, and N. J. White. 1995. In vitro antimicrobial susceptibilities of strains of Yersinia pestis. Antimicrob. Agents Chemother. 39:21532154.
61. Sookpranee, T.,, M. Sookpranee,, M. A. Mellencamp,, and L. C. Preheim. 1991. Pseudomonas pseudomallei, a common pathogen in Thailand that is resistant to the bactericidal effects of many antibiotics. Antimicrob. Agents Chemother. 35:484489.
62. Thibault, F. M.,, E. Hernandez,, D. R. Vidal,, M. Girardet,, and J. D. Cavallo. 2004. Antibiotic susceptibility of 65 isolates of Burkholderia pseudomallei and Burkholderia mallei to 35 antimicrobial agents. J. Antimicrob. Chemother. 54:11341138.
63. Trujillano-Martin, I.,, E. Garcia-Sanchez,, M. J. Fresnadillo,, J. E. Garcia-Sanchez,, and J. A. Garcia-Rodriguez. 1999. In vitro activities of five new antimicrobial agents against Brucella melitensis. Int. J. Antimicrob. Agents 12:185186.
64. Turnbull, P. C. B.,, N. M. Sirianni,, C. I. LeBron,, M. N. Samaan,, F. N. Sutton,, A. E. Reyes,, and L. F. Peruski. 2004. MICs of selected antibiotics for Bacillus anthracis, Bacillus cereus, Bacillus thuringiensis, and Bacillus mycoides from a range of clinical and environmental sources as determined by Etest. J. Clin. Microbiol. 42:36263634.
65.U. S. Department of Health and Human Services. 1999. Biosafety in microbiology and biomedical laboratories. J. Y. Richmond, and R. W. McKinney (ed.), U.S. Government Printing Office, Washington, D.C.
66. White, N. 2003. Meliodosis. Lancet. 361:17151722.
67. Working Party of the British Society for Antimicrobial Chemotherapy. 1988. Breakpoints in in-vitro antibiotic sensitivity testing. J. Antimicrob. Chemother. 21:701710.
68. Yamamoto, T.,, P. Naigowit,, S. Dejsirilert,, D. Chiewsilp,, E. Kondo,, T. Yokota,, and K. Kanai. 1990. In vitro susceptibilities of Pseudomonas pseudomallei to 27 antimicrobial agents. Antimicrob. Agents Chemother. 34:20272029.


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Table 1

Testing methods and antimicrobial agents to test and report for bioterrorism agents defined by CLSI/NCCLS

Reprinted from reference 10 with permission.

Citation: Tenover F. 2005. Antimicrobial Susceptibility Testing of Bacterial Agents of Bioterrorism: Strategies and Considerations, p 414-422. In White D, Alekshun M, McDermott P (ed), Frontiers in Antimicrobial Resistance. ASM Press, Washington, DC. doi: 10.1128/9781555817572.ch31

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