Background and Objectives: In December 2010 four, lions and one tiger died at the Tehran zoo. Out of all samples, Burkholderia mallei (causative agent of GLANDERS) was isolated just from ulcer sample of the tiger which was imported to Iran from Russia.Materials and Methods: One nasal swab from a tiger and fifteen blood samples with anticoagulant belonging to one tiger and fourteen lions (four dead lions and eleven live lions) were collected and were inoculated directly onto the selective media. The isolate was identified by morphological and biochemical and API BBL tests and PCR using specific primers (Bma-IS407-flip). The standard (Razi Type Culture Collection RTCC: 2375) and tiger isolates were inoculated into 2 guinea pigs. All residue solipeds and carnivores were checked by Malleination test and Complement Fixation (CF) Test respectively.Results: One isolate of B. mallei was isolated from tiger’s nasal swab. Both of B.mallei strains were recovered from inoculated animals. All of solipeds were negative by malleination test, however, 11 lions including 4 dead and 7 live lions out of 14 lions were positive in CF test for GLANDERS and all were put down by the authorities.Conclusion: Active surveillance of GLANDERS is essential for solipeds, especially it’s more important while being used to feed valuable carnivores like lions and tigers. Therefore, a reliable test like malleination must be carried out twice (first before transferring and one month after quarantine). Both test results should be negative for use for feeding.

Background: Death of one imported tiger from Russia in Tehran Eram zoological garden in winter of 2010 from GLANDERS opened the new series of discussion about this old zoonosis. After this incident, all the lions of the zoo were euthanized due to the probable contamination with GLANDERS.Materials and Methods: In present systematic review manuscript, several scientific data banks and websites including ISI, Scopus, Medline, Embase, OIE, CDC, WHO, SID and MAGIRAN had been searched. Also all the books, journals and available congress proceedings in libraries were searched about this issue.Results: As if many epidermis and also zoonotic and infectious diseases may be a biological threat, so having a good knowledge of these kinds of diseses is a necessity for all physicians, veterinarians and public health staffs especially whom they work in Armed Forces.Conclusion: Collaboration between Iran Veterinary Organization (IVO), Ministry of Health and Medical Education (MOHME) and Armed Forces is necessary for controlling the probable biological threat like GLANDERS.

GLANDERS is a zoonosis caused by Burkholderia mallei (B. mallei). GLANDERS has been re-emerging in recent years due to war in the Middle East, unauthorized transfer, the lack of formulated action plans, etc. The prevalence of GLANDERS in Iran and the risk of B. mallei transmission and infection is high, therefore requires the quick identify this disease in animals, particularly in horses. This study investigated GLANDERS re-emerging in horses in East Azerbaijan Province, Iran. From 22 September 2020 to 20 March 2021, six-month periodic tests such as the complement fixation test (CFT) were implemented by the Veterinary Administration of East Azerbaijan to detect GLANDERS in horses. In the case of positive CFT results, the mallein test was conducted. According to the test results, blood samples were taken to culture and prepare serum for the ELISA test. Deep swab samples were collected from nasal mucosa, lymph fluid, and blood. The CFT results indicated 12 horses were susceptible to GLANDERS, and three horses were diagnosed with GLANDERS based on the mallein confirmatory test results. B. mallei were not isolated in culturing the samples. Three cases were positive in the ELISA test which was consistent with the CFT and mallein test results. However, the molecular test results were negative. It is challenging to isolate B. mallei in the early stage of disease, and the negative molecular diagnostic test result may be misleading in GLANDERS diagnosis. In susceptible cases with a positive CF test result, GLANDERS can be diagnosed by skin mallein and ELISA tests.

Introduction and Objective: Burkholderia mallei, is the etiologic agent of the GLANDERS disease. Clinical and bacteriological diagnosis of GLANDERS is difficult in the early stages of the disease. Some methods such as Complement fixation test (CFT) due to false positive results and troublesome for veterinary authorities and cause financial losses to animal owners, the other one is Malleination test, which requires appropriate equipment and efficient laboratory personnel. Therefore, in order to quickly and accurately diagnose the disease, especially in areas that cannot be kept animals, new methods should be used to identify the disease. The Western blot is a serological diagnostic test and has been recommended by the World Organization for Animal Health (OIE). The aim of this study was to apply Western blot assay using purified lipopolysaccharide (LPS) containing antigen of Burkholderia mallei was designated. Material and Methods: In this study, a total of 75 sera were collected from different horse populations from several geographical areas of Iran. Specificity and sensitivity of the Complement fixation test, ELISA and a Western blot were compared for serodiagnosis of GLANDERS. ELISA test was based on B. mallei antigens and Western blot by using of a purified LPS-containing B. mallei-antigen. Results: The Western blot and ELISA were more specific than the Complement fixation test. ELISA based on B. mallei antigens had more sensitivity compared to Complement fixation test and Western blot. Finally, sensitivity and specificity were obtained for Complement fixation test (92. 31% and 98. 38%), Western blot (92. 31%, 100%) and (100% and 100%) respectively. Conclusion: Complement fixation test for GLANDERS is still the prescribed serological method for commercial purposes, which is used to confirm the health of animals. However, in order to maintain the biosafety of valuable and expensive horse colonies, it is important to implement a more accurate and intelligent disease control and eradication program. This enhances the laboratory diagnostic capabilities to better understand the cause of the disease and to use and optimize the diagnostic methods of GLANDERS in the country. Therefore, efforts to further improve and optimize ELISA and Western blotting should be continue.

In 27th of December 2010 a male Siberian tiger(Panthera tigris altaica) in Tehran zoo was visited and bilateral signs of acute systemic disease was noticed, epistaxis, inspiratory dispnea, polydipsia, coutaneus ulcers specially on tail, chest and limbs were recognized.After sampling and culture from nasal discharge an aerobic, gram negative, immotile and bipolar bacilli isolated.Clinical signs, characteristic of bacterial infection in addition to the history of feeding with donkey meat without performing Mallein test were our reason to become suspicious to GLANDERS that later was confirmed with PCR.Post mortem examination showed the multiple nasal mucosal nodules, mucosal erosions, cutaneous ulcers with fistula and sinus formation and yellowish discharge. The tiger was FIV positive too.

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).

Introduction GLANDERS is one of the oldest contagious and dangerous zoonotic diseases manifesting ulcerative granulomatous lesions on the skin and mucous membranes. Early methods possessing desirable sensitivity and specificity is important to diagnose the disease considering the just only one case report and preventing disease by identification and eradication. The present study was aimed to design and optimize Dot-blot ELISA system using specific antigens for simple and rapid detection of GLANDERS using equine sera samples. Materials & Methods Burkholderia mallei strains were cultured in nutrient broth supplemented with glycerol 4%. Whole cell antigens of bacterial strains were precipitated by trichloroacetic acid (TCA) followed by sonication method. The optimum concentrations of antigen-antibody were determined by checkerboard titration. The nitrocellulose membrane was coated by antigen, then stopped by skimmed milk as a blocker. The suspicious equine serum was added to the membrane following HRP-conjugated antibody, the signals were detected by TMB as a substrate. Results 4 out of 90 sera samples were positive by Dot-blot ELISA. The specificity and sensitivity of the test were evaluated 96. 62% and 100%, respectively. B. mallei culture and isolation from clinical specimens of the horses validated the test. Conclusion Our study showed that the Dot-blot ELISA is specific and sensitive in GLANDERS diagnosis and it seems to be practical and efficient test due to the rapid, easy interpretation without any special tools, cost-effective and also field-friendly.