Investigating the status of Brucella melitensis contamination and comparing different molecular and non-molecular diagnosis methods in one of Kennel dogs in suburb of Tehran- Iran

Document Type : Research Paper

Authors

1 Associate Professor, Department of Theriogenology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

2 DVM Graduated, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

3 Professor, Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

4 Expert, Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

10.22055/ivj.2023.387783.2563

Abstract

    Brucellosis caused by Brucella melitensis is one of the important health and economic diseases in Iran is very important to pay attention to it as a zoonotic disease. The aim of the study was to investigate B. melitensis infection in a kennel dog and also to compare different laboratory methods to diagnose Brucella melitensis infection in dogs. In this study, the whole blood samples (for PCR and culture), vaginal swabs samples (for PCR tests) and serum samples (for Rose Bengal, Wright and 2 mercaptoethanol tests) were obtained from 14 bitches at a kennel, in which a bitch had been aborted due to Brucella melitensis infection. Also, the whole blood sample and semen sample were obtained from the only male dog of this kennel for PCR test. Finally, the data obtained from these tests were analyzed by calculating the sensitivity and specificity as well as the Kappa coefficient. Bacteriological culture results of all blood samples were negative. The PCR results of seven blood samples (%50) and eight vaginal swab samples (%57.1) obtained from 14 bitches were positive. By using PCR primers, B. melitensis were identified in all positive samples. In the serological test, out of 14 sera obtained from bitches for the Rose Bengal screening test, eight positive samples were observed, but only one positive sample was observed in Wright and 2 mercaptoethanol tests. The results showed that the sensitivity and specificity of PCR test in vaginal swab samples was 100% and there is an excellent performance (kappa coefficient = 1) between the results of the Rose Bengal test and PCR of vaginal swab samples in the diagnosis of Brucella melitensis in dogs. In dogs suspected of brucellosis, at first it is recommended to take a sample of the animal's blood to perform the Rose Bengal test, and if the result is positive, to confirm the diagnosis and to determine the strain of Brucella, then a vaginal swab sample should be taken to perform the PCR test.

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Main Subjects

References

Al-Garadi, M. A., Khairani-Bejo, S., Zunita, Z., Omar, A. R. (2011). Detection of Brucella melitensis in blood samples collected from goats. Journal of Animal and Veterinary Advances, 10(11), 1437-1444. doi: 10.3923/javaa.2011.1437.1444
Al Jindan, R. (2021). Scenario of pathogenesis and socioeconomic burden of human brucellosis in Saudi Arabia. Saudi Journal of Biological Sciences, 28(1), 272-279. doi: 10.1016/j.sjbs.2020.09.059
Alamian, S., & Dadar, M. (2020). Brucella melitensis infection in dog: a critical issue in the control of brucellosis in ruminant farms. Comparative Immunology, Microbiology and Infectious Diseases, 73, 101554. doi: 10.1016/j.cimid.2020.101554
Alikhani, M. Y., Hashemi, S. H., Naseri, Z., Farajnia, S., Peeri, D. H. (2013) Diagnosis of human brucellosis by blood culture (BACTEC) and PCR method via whole blood and serum. Jundishapur Journal of Microbiology, 6(3), 248-251. doi: 10.5812/jjm.5073
Amin, A. S., Hamdy, M. E., & Ibrahim, A. K. (2001). Detection of Brucella melitensis in semen using the polymerase chain reaction assay. Veterinary microbiology, 83(1), 37-44. doi: 10.1016/s0378-1135(01)00401-1
Beehan D. P. (2017) Brucellosis. In S. J, Ettinger, E. C, Feldman, E, Cote (Eds.). Textbook of Veterinary Internal Medicine (8th ed., pp. 2290-2294). Elsevier Saunders. Philadelphia, USA.
Dadar, M., Alamian, S., Behrozikhah, A. M., Yazdani, F., Kalantari, A., Etemadi, A., & Whatmore, A. M. (2019). Molecular identification of Brucella species and biovars associated with animal and human infection in Iran. Veterinary research forum, 10(4), 315-321. doi: 10.30466/VRF.2018.89680.2171
Davidson A. P. (2019) The practice of theriogenology. In R. W, Nelson, C. G, Couto (Eds.). Small animal internal medicine (6th ed., pp. 935-952). Elsevier. Amsterdam.
Dehkordi, F. S., Khamesipour, F., & Momeni, M. (2014). Brucella abortus and Brucella melitensis in Iranian bovine and buffalo semen samples: The first clinical trial on seasonal, Senile and geographical distribution using culture, conventional and real-time polymerase chain reaction assays. Kafkas Universitesi Veteriner Fakultesi Dergisi, 20(6), 821-828. doi: 10.9775/kvfd.2014.10827
Esmaeili, H., Mahdavi, A. S, Hamedi, M. (2021). Efficacy of Rev-1 Vaccine Against Brucella melitensis Infection in Dog. Iranian Journal of Veterinary Medicine, 15(4), 387-394. doi: 10.22059/IJVM.2021.309899.1005126
Esmaeili, H. (2014). Brucellosis in Islamic republic of Iran. Journal of medical bacteriology, 3(3-4), 47-57.
Gall, D., & Nielsen, K. (2004). Serological diagnosis of bovine brucellosis: a review of test performance and cost comparison. Revue scientifique et technique, 23(3), 989-1002. doi: 10.20506/rst.23.3.1545
Gharekhani, J., & Sazmand, A. (2019). Detection of Brucella antibodies in dogs from rural regions of Hamedan, Iran. Avicenna Journal of Clinical Microbiology and Infection, 6(4), 122-126. doi: 10.34172/ajcmi.2019.22
Ghorbani, A., Rabbani Khorasgani, M., Zarkesh-Esfahani, H., Sharifiyazdi, H., Kashani, A. D., & Emami, H. (2013). Comparison of serology, culture, and PCR for detection of brucellosis in slaughtered camels in Iran. Comparative Clinical Pathology, 22, 913-917. doi: 10.1007/s00580-012-1499-1
Golshani, M., & Buozari, S. (2017). A review of brucellosis in Iran: epidemiology, risk factors, diagnosis, control, and prevention. Iranian biomedical journal, 21(6), 349. doi: 10.18869/acadpub.ibj.21.6.349
Gupta, V. K., Verma, D. K., Rout, P. K., Singh, S. V., & Vihan, V. S. (2006). Polymerase chain reaction (PCR) for detection of Brucella melitensis in goat milk. Small Ruminant Research, 65(1-2), 79-84. doi: 10.1016/j.smallrumres.2005.05.024
Hajia, M. (2018). The Challenges in Diagnosis of Brucellosis Serological Tests and Available Approaches. Iranian Journal of Medical Microbiology, 12(1), 1-5.
Hollett, R. B. (2006). Canine brucellosis: outbreaks and compliance. Theriogenology, 66(3), 575-587. doi: 10.1016/j.theriogenology.2006.04.011
Ilhan, Z., Boynukara, B., Solmaz, H., Ekin, I. H., Aksakal, A., & Gulhan, T. (2008). Detection of Brucella melitensis DNA in the milk of sheep after abortion by PCR assay. Archivos de medicina veterinaria, 40(2), 141-146. doi: 10.4067/S0301-732X2008000200005
Jamil, T., Melzer, F., Khan, I., Iqbal, M., Saqib, M., Hammad Hussain, M., ... & Neubauer, H. (2019). Serological and molecular investigation of Brucella species in dogs in Pakistan. Pathogens, 8(4), 294. doi: 10.3390/pathogens8040294
Kaushik, P., Singh, D. K., Tiwari, A. K., & Kataria, R. S. (2006). Rapid detection of Brucella species in cattle semen by PCR. Journal of Applied Animal Research, 30(1), 25-28. doi: 10.1080/09712119.2006.9706818
Keid, L. B., Soares, R. M., Vasconcellos, S. A., Chiebao, D. P., Salgado, V. R., Megid, J., & Richtzenhain, L. J. (2007a). A polymerase chain reaction for detection of Brucella canis in vaginal swabs of naturally infected bitches. Theriogenology, 68(9), 1260-1270. doi: 10.1016/j.theriogenology.2007.08.021
Keid, L. B., Soares, R. M., Vasconcellos, S. A., Chiebao, D. P., Megid, J., Salgado, V. R., & Richtzenhain, L. J. (2007b). A polymerase chain reaction for the detection of Brucella canis in semen of naturally infected dogs. Theriogenology, 67(7), 1203-1210. doi: 10.1016/j.theriogenology.2007.01.003
Keid, L. B., Soares, R. M., Vasconcellos, S. A., Megid, J., Salgado, V. R., & Richtzenhain, L. J. (2009). Comparison of agar gel immunodiffusion test, rapid slide agglutination test, microbiological culture and PCR for the diagnosis of canine brucellosis. Research in veterinary science, 86(1), 22-26. doi: 10.1016/j.rvsc.2008.05.012
Kim, S., Lee, D. S., Suzuki, H., & Watarai, M. (2006). Detection of Brucella canis and Leptospira interrogans in canine semen by multiplex nested PCR. Journal of Veterinary Medical Science, 68(6), 615-618. doi: 10.1292/jvms.68.615
Lingam, G. S., Kumar, A. V., Singh, S., Kumar, M. S., & Krishnaiah, N. (2020). Sero occurence of brucellosis in dogs of Telangana state. Journal of Pharmaceutical Innovation, 9, 452-455.
Lopate, C. (2017) Vaginoscopy and vaginal cytology in Dogs. In S. J, Ettinger, E. C, Feldman, E, Cote (Eds.). Textbook of Veterinary Internal Medicine (8th ed., pp. 1263-1269). Elsevier Saunders. Philadelphia, USA.
McKinnon, A. O., Squires, E. L., Vaala, W.E., Varner, D. D. (2011). Equine Reproduction (2nd ed.). Wiley-Blackwell. New Jersey, USA.
Megid, J., Mathias, L. A., & Robles, C. A. (2010). Clinical manifestations of brucellosis in domestic animals and humans. The Open Veterinary Science Journal, 4, 119-126. doi: 10.2174/1874318801004010119
Mol, J. P., Guedes, A. C., Eckstein, C., Quintal, A. P., Souza, T. D., Mathias, L. A., ... & Santos, R. L. (2020). Diagnosis of canine brucellosis: comparison of various serologic tests and PCR. Journal of Veterinary Diagnostic Investigation, 32(1), 77-86. doi: 10.1177/1040638719891083
Quinn, P. J., Markey, B. K., Leonard, F. C., Hartigan, P., Fanning, S., & Fitzpatrick, E. (2011). Veterinary microbiology and microbial disease (2nd ed.). Wiley-Blackwell. New Jersey, USA.
Rezaei-Sadaghiani, R., Zowghi, E., Marhemati-Khamene, B., Mahpeikar, H. A. (1996). Brucella melitensis infection in sheep-dogs in Iran. Arch Razi Inst, 46(47), 1-7. doi: 10.22092/ARI.1996.109149
Sadeghi, H., Ashrafi Tamai, I., Vodjgani, M., Gharagozlou, F., Zahraei Salehi, T. (2022). Isolation and Identification of Brucella melitensis Biovar 1 using Bacteriological, Serological, and Molecular Tools from Saanen Goats (Capra aegagrus hircus) in Alborz, Iranian Journal of Veterinary Research, 77(2), 107-115. doi: 10.22059/jvr.2021.318798.3133
Sam, I. C., Karunakaran, R., Kamarulzaman, A., Ponnampalavanar, S., Omar, S. S., Ng, K. P., ... & AbuBakar, S. (2012). A large exposure to Brucella melitensis in a diagnostic laboratory. Journal of Hospital Infection, 80(4), 321-325. doi: 10.1016/j.jhin.2011.12.004
Santos, R. L., Souza, T. D., Mol, J. P., Eckstein, C., & Paíxão, T. A. (2021). Canine brucellosis: an update. Frontiers in Veterinary Science, 8, 594291. doi: 10.3389/fvets.2021.594291
Sharma, D., Bhardwaj, B., Chouhan, H., Chandel, B., Joseph, B., & Barolia, S. (2018). Isolation and molecular characterization of Brucella melitensis from abortion storms in sheep with its zoonotic implications. International Journal of Current Microbiology and Applied Sciences, 7(10), 1348-1356. doi: 10.20546/ijcmas.2018.710.150
Talebkhan-Garoussi, M., Mehrzad, J., Baniassadi, A., Khoshnegah, J. (2018). Seroprevalence of brucellosis in different kinds of feline population in north-east of Iran. Comp Clin Pathol, 27, 1155–1160. doi: 10.1007/s00580-018-2714-5
Talebkhan Garroussi, M. Firoozi, S., Nowrozian. I. (1997). The serological survey of Brucella abortus and melitensis in shepherd dogs around Mashhad farms. Journal of Faculty of Veterinary Medicine. University of Tehran. (Abstract in English). 51, 3&4: 63-72
Volkweis, F. S., Cavalcanti, L. C., Blume, H., Júnior, H. L. S., Lazzari, A., & Mulinari, F. (2018). Detection of Brucella canis in blood, urine and seminal fluid of a naturally infected dog by PCR. Brazilian Journal of Veterinary Medicine, 40(1), e002718-e002718. doi: 10.29374/2527-2179.bjvm002718
Wallach, J. C., Giambartolomei, G. H., Baldi, P. C., & Fossati, C. A. (2004). Human infection with M-strain of Brucella canis. Emerging infectious diseases, 10(1), 146. doi: 10.3201/eid1001.020622
Wanke, M. M. (2004). Canine brucellosis. Animal reproduction science, 82, 195-207. doi: 10.1016/j.anireprosci.2004.05.005
Zhu, X., Zhao, Z., Ma, S., Guo, Z., Wang, M., Li, Z., & Liu, Z. (2020). Brucella melitensis, a latent “travel bacterium,” continual spread and expansion from Northern to Southern China and its relationship to worldwide lineages. Emerging Microbes & Infections, 9(1), 1618-1627. doi: 10.1080/22221751.2020.1788995

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