Detection And Molecular Identification Of Burkholderia mallei Isolated From Blood Specimen Of An Infected Horse With Glanders

Document Type : Full Research Paper

Authors

1 Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Department of Tuberculin and Mallein, Razi Vaccine & Serum Research Institute, Agricultural Research, Education and Extension Organization, Karaj, Iran

3 Veterinary Aerobic Bacteria Vaccines Department, Razi Vaccine & Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran

Abstract

Burkholderia mallei is a causative agent of glanders, a zoonotic disease and one of the most dangerous and oldest contagious diseases in equidae. Iran has still been recognized as the major center for glanders. The present study was conducted to isolate B. mallei from blood specimen of an infected horse with glanders reported from Oshnavieh-West Azerbaijan in the past decade and molecular identification based on specific genes BimA, IS407-flip, and 23S rRNA.The blood sample of infected horse were cultured in biphasic medium containing nutrient broth and nutrient agar supplemented with glycerin and antibiotic. The bacterial isolate was identified by biochemical tests. The isolate was inoculated to male guinea pig intraperitoneally as a sensitive host for pathobiological studies. B. mallei isolate was verified by PCR and sequencing of BimA, IS407-flip, and 23S rRNA genes. B. mallei was isolated from blood sample of infected horse. The major sign of testicular swelling was seen in the male guinea pig after about 72 h IP inoculation with B. mallei and testicular colonization. The PCR amplification of BimA, IS407-flip, and 23S rRNA genes of B. mallei resulted in expected sizes of 989 bp, 250 bp, and 526 bp, respectively. Burkholderia pseudomallei and Pseudomonas aeruginosa were used as negative control.The report of glanders is alarming for healthcare organizations and need to monitor carefully. Due to the complication of diagnosis of glanders infectious agent, the identification of B. mallei using BimA, IS407-flip, and 23S rRNA is according to the diagnostic standards of World Organisation for Animal Health (OIE).

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References

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Veterinary Research & Biological Products
Volume 32, Issue 2
July 2019
Pages 2-10

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History

  • Receive Date: 29 July 2018
  • Revise Date: 06 September 2018
  • Accept Date: 25 September 2018

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