Early Detection of Bacillus anthracis From Saliva in Anticipation of a Bioterrorism Attack

Tigor Rona Airlangga Harya Bima, Rahmat Setya Adji, Elza Ibrahim Auerkari


Objective:To assess potential for early detection of oral infection by B. anthracisspores for preparedness of a bioterrorism attack. Material and Methods:The laboratory study used saliva with a range of initial anthrax concentrations, to compare detection by direct observation from conventional blood agar culture and by anthrax-specific PCR after a shorter culture in BHI broth. Three types of saliva were collected: stimulated saliva, unstimulated/whole saliva, and unstimulated/whole saliva with antibiotic treatment (for negative control). Using bivariate Kruskal-Wallis and Mann-Whitney tests for statistical analysis for factors that could affecting anthrax detection, significant differences between the test groups was assumed at p<0.05. Results:From unstimulated whole saliva heat shock treated at 62.50C, B. anthracisgrowth was detected with both methods. PCR detection from a BHI broth culture could shorten the time to diagnosis in comparison to conventional culture in blood agar. Conclusion:Saliva can provide useful samples for diagnosis of oropharyngeal anthrax. In comparison to conventional culture on blood agar, shorter-term culture in BHI broth provides potential for earlier detection and diagnosis.


Bioterrorism; Biological Warfare; Anthrax; Saliva

Full Text:



Pal M, Tsegaye M, Girzaw F, Bedada H, Godishala V, Kandi V. An overview on biological weapons and bioterrorism. Am J Biomed Res 2017; 5(2):24-34. https://doi.org/10.12691/ajbr-5-2-2

Goel AK. Anthrax: A disease of biowarfare and public health importance. World J Clin Cases 2015; 3(1):20-33. https://doi.org/10.12998/wjcc.v3.i1.20

Doganay M, Demiraslan H. Human anthrax as a re-emerging disease. Recent Pat Antiinfect Drug Discov 2015; 10(1):10-29. https://doi.org/10.2174/1574891X10666150408162354

National Center for Emerging and Zoonotic Infectious Diseases (NCEZID). Centers for Disease Control and Prevention. Guide to Understanding Anthrax. Available at: https://www.cdc.gov/anthrax/pdf/evergreen-pdfs/anthrax-evergreen-content-english.pdf. [Accessed on October 14, 2018].

World Health Organization. Anthrax in Humans and Animals. 4th. ed. Geneva: World Health Organization, 2008. 208p.

Friend M, Hurley JW, Nol P, Wesenberg K. Biowarfare, Bioterrorism, and Animal Diseases as Bioweapons. In: Friend M, Hurley JW, Nol P, Wesenberg K. Disease Emergency and Resurgence: The Wildlife-Human Connection. Virginia: US Geological Survey, 2006. Chapter 6. p. 234-265.

Washington State Department of Health. Anthrax: Reporting and Surveillance Guidelines. Available at: https://www.doh.wa.gov/Portals/1/Documents/5100/420-045-Guideline-Anthrax.pdf. [Accessed on October 14, 2018].

Bienek DR, Chang CK, Nieva CD, Slowey PD. Detection of protective antigen-spesific antibodies in saliva of anthrax-vaccinated individual. Int Assoc Dent Res 2004. Abstract #1831.

Bienek DR, Chang CK, Cohen ME. Detection of anti-protective antigen salivary IgG antibodies in recipients of the US licensed anthrax vaccine. Vaccine 2007; 25(32):5978-84. https://doi.org/10.1016/j.vaccine.2007.05.043

Pfaffe T, Cooper-White J, Beyerlein P, Kostner K, Punyadeera C. Diagnostic potential of saliva: Current state and future applications. Clin Chem 2011; 57(5):675-87. https://doi.org/10.1373/clinchem.2010.153767

Yunanto A, Chandra MS, Widjajanto E, Widodo MA. Quantity, quality, and neutrophils phagocytosis in saliva and blood of newborn babies with sepsis risk. J Kedokteran Brawijaya 2012; 27(2):89-95.

Gomar-Vercher S, Simon-Soro A, Montiel-Company JM, Almerich-Sila JM, Mira A. Stimulated and unstimulated saliva samples have significantly different bacterial profiles. PLoS One 2018; 13(6):e0198021. https://doi.org/10.1371/journal.pone.0198021

Belstrøm D, Holmstrup P, Bardow A, Kokaras A, Fiehn N, Paster BJ. Comparative analysis of bacterial profiles in unstimulated and stimulated saliva samples. J Oral Microbiol 2016; 8. https://doi.org/10.3402/jom.v8.30112

Belstrøm D, Fiehn NE, Nielsen CH, Klepac-Ceraj V, Paster BJ, Twetman S, et al. Differentiation of salivary bacterial profiles of subjects with periodontitis and dental caries. J Oral Microbiol 2015; 7:27429. https://doi.org/10.3402/jom.v7.27429