Veterinary Research Forum
Vet Res Forum

Isolation, serological and molecular methods in screening of Burkholderia mallei in East Azerbaijan province, Iran

Document Type : Original Article

Authors

Hassan Tizfahm Tikmehdash 1
Alireza Dehnad 2
Nader Mosavari 3
Behroz Naghili Hokmabadi 4
Sanaz Mahmazi 1

1 Department of Biology, Faculty of Basic Sciences, Islamic Azad University, Zanjan Branch, Zanjan, Iran

2 Department of Livestock Bacterial Diseases Research, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization, Karaj, Iran

3 Reference Laboratory of Bovine Tuberculosis, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization, Karaj, Iran

4 Infectious and Tropical Diseases Research Centre, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran

https://doi.org/10.30466/vrf.2024.2010651.4002
Abstract
Glanders caused by Burkholderia mallei is one of the most dangerous zoonotic diseases in solipeds. Clinical diagnosis of this disease in its early stages in horses, is difficult. This study investigated serological and molecular identification of B. mallei in East Azerbaijan province. In the third and fourth quarters of 2020, throughout 2021, and in the first and second quarters of 2022, the complement fixation test (CFT) was performed on 350 horses. The malleination was used to confirm the positive CFT cases. Blood samples were taken for culture and for preparing serums to perform the enzyme-linked immunosorbent assay (ELISA). Deep eye discharge, nostril, cutaneous ulcers and lymph fluid swabs were cultured, and polymerase chain reaction (PCR) was carried out. Eleven horses were CFT-positive. Based on the malleination on the 11 horses, six were affected by glanders, five were not affected (false positive), and one horse was CFT-negative despite exhibiting clinical signs. It was positive by malleination, ELISA and PCR. A total number of seven positive cases of glanders were diagnosed. The B. mallei could not be isolated, but the Burkholderia cepacia complex was isolated in one case. Except for three cases, the results of the CFT, mallein and ELISA tests were consistent. The amount of confidence interval was 95.00%. It is suggested that ELISA could be used as a complement to CFT in screening and, if positive results are observed in one of the tests, the entire herd must be examined more accurately using the mallein and western blot confirmatory tests.

Keywords

Burkholderia mallei
Complement fixation test
ELISA
Glanders
Malleination

Subjects

  1. Van Zandt KE, Greer MT, Gelhaus HC. Glanders: an overview of infection in humans. Orphanet J Rare Dis 2013; 8: 131.doi: 10.1186/1750-1172-8-131.
  2. Khaki P, Mosavari N, Khajeh-Nasiri S, et al. Glanders outbreak at Tehran zoo, Iran. Iran J Med Microbiol 2012; 4(1): 3-7.
  3. Nguyen HN, Smith ME, Hayoun MA. Glanders and melioidosis. StatPearls2023. Available at: https://www.ncbi.nlm.nih.gov/books/NBK448110/. Accessed 02 April 2024.
  4. Inglis TJJ, Merritt AJ. Burkholderia pseudomallei and Burkholderia mallei. In: Tang YW, Sussman M, Liu D, et al. (Eds). Molecular medical microbiology. 2nd Massachusetts, USA: Academic Press 2014; 769-791.
  5. LipsitzR, Garges S, Aurigemma R, et al. Workshop on treatment of and postexposure prophylaxis for Burkholderia pseudomallei and Burkholderia mallei infection, 2010. Emerg Infect Dis 2012; 18(12): e2.doi: 10.3201/eid1812.120638.
  6. Yazdansetad S, Mosavari N, Tadayon K, et al. Development of an immunoblotting assay for serodiagnosis of Burkholderia mallei infection: the whole-cell proteome-based paradigm. Iran J Microbiol 2019; 11(3): 232-238.
  7. Yazdansetad S, Mosavari N, Tadayon K, et al. Detection and molecular identification of Burkholderia mallei isolated from blood specimen of an infected horse with glanders [Persian]. Vet Res Biol Prod 2018; 123: 2-10.
  8. Estes DM, Dow SW, Schweizer HP, et al. Present and future therapeutic strategies for melioidosis and glanders. Expert Rev Anti Infect Ther 2010; 8(3): 325-338.
  9. Tikmehdash HT, Dehnad A, Mosavari N, et al. Glanders reemerging in few horses in East-Azerbaijan, Iran. J Zoonotic Dis 2022; 6(2): 60-68.
  10. World Organization for Animal Health. Glanders, Chapter 3.5.11. Paris, France: OIE Terrestrial Manual; 2018.
  11. Falcão MVD, Laroucau K, Vorimore F, et al. Molecular characterization of Burkholderia mallei strains isolated from horses in Brazil (2014-2017). Infect Genet Evol 2022; 99: 105250.doi: 10.1016/j.meegid.2022.105250.
  12. Abreu DC, Gomes AS, Tessler DK, et al. Systematic monitoring of glanders-infected horses by complement fixation test, bacterial isolation, and PCR. Vet Anim Sci 2020; 10: 100147.doi: 10.1016/j.vas.2020.100147.
  13. OIE Terrestrial Manual. Glanders, Chapter 2. 5. 11. Paris, France: World Organization for Animal
    Health; 2013.
  14. Ulrich RL, Ulrich MP, Schell MA, et al. Development of a polymerase chain reaction assay for the specific identification of Burkholderia mallei and differentiation from Burkholderia pseudomallei and other closely related Burkholderiaceae. Diagn Microbiol Infect Dis 2006; 55(1): 37-45.
  15. Nasiri M, Zarrin A, RoshankarRudsari S, et al. Glanders (Burkholderia mallei infection) in an Iranian man: a case report. IDCases 2023; 32: e01779.doi: 10.1016/ j.idcr.2023.e01779.
  16. Dashtipour S, Tadayon K, Yazdansetad S, et al. Genomic pattern analysis of Burkholderia mallei field isolates by pulsed-field gel electrophoresis (PFGE) discriminatory typing. Iran J Microbiol 2021; 13(5): 574-582.
  17. Shakibamehr N, Mosavari N, Harzandi N, et al. Designing of western blot technique for glanders diagnosing in Iran. J Equine Vet Sci 2021; 99: 103403.doi: 10.1016/j.jevs.2021.103403.
  18. Laroucau K, Colaneri C, Jaÿ M, et al. Interlaboratory ring trial to evaluate CFT proficiency of European laboratories for diagnosis of glanders in equids. Vet Rec 2016; 178(25): 632. doi: 10.1136/vr.103617.
  19. Elschner MC, Melzer F, Singha H, et al. Validation of a commercial glanders ELISA as an alternative to the CFT in international trade of equidae. Front Vet Sci 2021; 8: 628389. doi: 10.3389/fvets.2021.628389.
  20. Laroucau K, Aaziz R, Vorimore F, et al. A genetic variant of Burkholderia mallei detected in Kuwait: consequences for the PCR diagnosis of glanders. Transbound Emerg Dis 2021; 68(2): 960-963.
  21. Mosavari N, Dehnad A, Naghili B, et al. Serum investigation, isolation and molecular identification of pathogenic Burkholderia from solipeds and respiratory secretions of cystic fibrosis patients in East Azarbaijan province. Agricultural research, education and extension organization (AREEO), Razi vaccine and serum research institute. Research project with approved code 34-18-1851-017-000176.
  22. Singha H, Shanmugasundaram K, Tripathi BN, et al. Serological surveillance and clinical investigation of glanders among indigenous equines in India from 2015 to 2018. Transbound Emerg Dis 2020; 67(3): 1336-1348.
  23. DeShazer D, Waag DM, Fritz DL, et al. Identification of a Burkholderia mallei polysaccharide gene cluster by subtractive hybridization and demonstration that the encoded capsule is an essential virulence determinant. Microb Pathog 2001; 30(5): 253-269.
  24. Kumar S, Malik P, Verma SK, et al. Use of a recombinant Burkholderia intracellular motility A protein for immunodiagnosis of glanders. Clin Vaccine Immunol 2011; 18(9): 1456-1461.
  25. Waag DM, England MJ, DeShazer D. Humoral immune responses in a human case of glanders. Clin Vaccine Immunol 2012; 19(5):814-816.
Veterinary Research Forum
Volume 15, Issue 5
May 2024
Pages 231-236

  • Receive Date 02 September 2023
  • Revise Date 03 December 2023
  • Accept Date 08 January 2024

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