Veterinary Research Forum
Vet Res Forum

Molecular identification of Brucella species and biovars associated with animal and human infection in Iran

Document Type : Original Article

Authors

Maryam Dadar 1
Saeed Alamian 1
Ali Mohammad Behrozikhah 1
Freshteh Yazdani 1
Armin Kalantari 1
Afshar Etemadi 1
Adrian M. Whatmore 2

1 Department of Brucellosis, Razi Vaccine and Serum Research Institute (RVSRI); Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran

2 Animal and Plant Health Agency (APHA), Addlestone, Surrey, United Kingdom

https://doi.org/10.30466/vrf.2018.89680.2171
Abstract
Brucellosis is a costly contagious disease of human, domestic and wild animals. It is a serious health problem in Iran causing significant economic losses therefore, control approaches to prevent its spread are of great importance. In Iran, the species and biovars of virulent Brucella species are still under-reported due to the inadequate diagnostic protocols and insufficient laboratory facilities. The objective of this study was to characterize Brucella isolates obtained from passive animal and human surveillance in Iran from 2011 to 2018 in order to understand the current epidemiological situation of the disease. A total of 419 samples (milk, blood, cerebrospinal fluid, abomasum content and aborted fetus tissues) were collected from 65 cases/case series (human and animals) and examined bacteriologically. The initially identified Brucella isolates were further characterized using phenotypic and molecular approaches. All recovered isolates were either B. abortus or B. melitensis. The infection in sheep appeared to be exclusively associated with B. melitensis, but both B. abortus and B. melitensis were common in bovine samples. Samples from one sheep and one goat were confirmed to be infected by the B. melitensis vaccine strain Rev1. In spite of B. abortus burden in animals (14 cases in cattle and camel), brucellosis in human was predominantly associated with B. melitensis (15 cases). The results confirmed that B. melitensis biovar 1 and B. abortus biovar 3 remain the most prevalent biovars in Iran. This report builds a picture of the significance of different Brucella species in different hosts in Iran and provides applicable information for the healthcare professionals about the public health risks of brucellosis and relevant preventive strategies.

Keywords

Brucellosis
Bruce ladder
PCR
Brucella species and biovars

Subjects

  1.  

    1. Cutler S, Whatmore A, Commander N. Brucellosis–new aspects of an old disease. J Appl Microbiol 2005; 98(6):1270-1281.
    2. Kolo FB, Fasina FO, Ledwaba B, et al. Isolation of Brucella melitensis from cattle in South Africa. Vet Rec 2018;182(23):668-669.
    3. Seleem MN, Boyle SM, Sriranganathan N. Brucellosis: a re-emerging zoonosis. Vet Microbiol 2010; 140(3-4): 392-398.
    4. Ezama A, Gonzalez JP, Majalija S, et al. Assessing short evolution brucellosis in a highly brucella endemic cattle keeping population of Western Uganda: A complementary use of Rose Bengal test and IgM rapid diagnostic test. BMC public health 2018;18(1):315. doi: 10.1186/s12889-018-5228-9.
    5. Shoukat S, Wani H, Ali U, et al. Brucellosis: A current review update on zoonosis. J Immunol Immunopathol 2017;19(2):61-69.
    6. Zhong Z, Yu S, Wang X, et al. Human brucellosis in the People's Republic of China during 2005–2010. Int J Infect Dis 2013;17(5):e289-e292.
    7. Sofian M, Aghakhani A, Velayati AA, et al. Risk factors for human brucellosis in Iran: A case–control study. Int J Infect Dis 2008;12(2):157-161.
    8. Behzadnia S, Najafi N, Marzban K, et al. Evaluation of epidemiologic, clinical, and paraclinical features of children with brucellosis hospitalized in two teaching hospital related to Mazandaran University of Medical Sciences, Iran, during the years 2010-2016. Chron Dis J 2018;5(1):12-18.
    9. Mirnejad R, Jazi FM, Mostafaei S, et al. Epidemiology of brucellosis in Iran: A comprehensive systematic review and meta-analysis study. Microb Pathog 2017; 109: 239-247.
    10. Roushan MH, Mohrez M, Gangi SS, et al. Epidemio-logical features and clinical manifestations in 469 adult patients with brucellosis in Babol, Northern Iran. Epidemiol Infect 2004;132(6):1109-1114.
    11. Tahamtan Y, Namavari MM, Mohammadi G, et al. Prevalence of brucellosis in horse north-east of Iran. J Equine Vet Sci 2010;30(7):376-378.
    12. Esmaeili H. Brucellosis in Islamic republic of Iran. J Med Bacteriol 2015; 3(3-4):47-57.
    13. Farahani S, Shah-mohamadi S, Navidi I, et al. An investigation of the epidemiology of brucellosis in Arak City, Iran (2001-2010). J Arak Uni Med Sci 2012; 14(7): 49-54.
    14. Sofian M, Sheikholeslami M, Mahdaviani FA, et al. Low prevalence of brucella agglutinins in blood donors in central province of Iran. Iran J Microbiol 2013; 5(1):24-27.
    15. Eini P, Keramat F, Hasanzadehhoseinabadi M. Epidemiologic, clinical and laboratory findings of patients with brucellosis in Hamadan, west of Iran. J Res Health Sci 2012; 12(2):105-108.
    16. Kassiri H, Amani H, Lotfi M. Epidemiological, laboratory, diagnostic and public health aspects of human brucellosis in western Iran. Asian Pac J Trop Biomed 2013; 3(8):589-594.
    17. Beheshti S, Rezaian G, Azad F, et al. Seroprevalence of brucellosis and risk factors related to high risk occupational groups in Kazeroon, South of Iran. Int J Occup Environ Med 2010; 1(2): 62-68.
    18. Alavi SM, Rafiei A, Nikkhooi A. The effect of lifestyle on brucellosis among nomads in Khuzestan province of Iran. Pak J Med Sci 2007; 23(3):358-360.
    19. Bokaie S, Sharifi L, Alizadeh H. Epidemiological survey of brucellosis in human and animals in Birjand, east of Iran. J Anim Vet Adv 2008; 7(4):460-463.
    20. Dean AS, Crump L, Greter H, et al. Global burden of human brucellosis: A systematic review of disease frequency. PLoS Negl Trop Dis. 2012; 6(10):e1865. doi: 10.1371/journal.pntd.0001865.
    21. Zowghi E, Ebadi A, Yarahmadi M. Isolation and identification of Brucella organisms in Iran. IranJ Clin Infec Dis 2008; 3(4):185-188.
    22. Behroozikhah AM, Bagheri Nejad R, Amiri K, et al. Identification at biovar level of Brucella isolates causing abortion in small ruminants of Iran. J Pathog 2012; 357235. doi: 10.1155/2012/357235.
    23. Alton G, Jones L, Angus R, et al. Techniques for the Brucellosis laboratory. Paris, France: Institute National de la Recherdie Agrononique 1988; page number?
    24. Godfroid J, Garin-Bastuji B, Saegerman C, et al. Brucellosis in terrestrial wildlife. Rev Sci Tech 2013; 32(1): 27-42.
    25. Probert WS, Schrader KN, Khuong NY, et al. Real-time multiplex PCR assay for detection of Brucella spp., B. abortus, and B. melitensis. J Clin Microbiol 2004; 42(3):1290-1293.
    26. Song L, Li J, Hou S, et al. Establishment of loop-mediated isothermal amplification (LAMP) for rapid detection of Brucella spp. and application to milk and blood samples. J Microbiol Methods 2012; 90(3):
      292-297.
    27. Ewalt DR, Bricker BJ. Validation of the abbreviated Brucella AMOS PCR as a rapid screening method for differentiation of Brucella abortus field strain isolates and the vaccine strains, 19 and RB51. J Clin Microbiol 2000; 38(8):3085-3086.
    28. López-Goñi I, García-Yoldi D, Marin C, et al. Evaluation of a multiplex PCR assay (Bruce-ladder) for molecular typing of all Brucella species, including the vaccine strains. J Clin Microbiol 2008; 46(10):3484-3487.
    29. Bricker BJ, Halling SM. Differentiation of Brucella abortus bv. 1, 2, and 4, Brucella melitensis, Brucella ovis, and Brucella suis bv. 1 by PCR. J Clin Microbiol 1994; 32(11):2660-2666.
    30. El-Diasty M, Wareth G, Melzer F, et al. Isolation of Brucella abortus and Brucella melitensis from seronegative cows is a serious impediment in Brucellosis control. Vet Sci 2018; 5(1): E28. doi: 10.3390/vetsci5010028.
    31. Musallam I, Abo-Shehada M, Hegazy Y, et al. Systematic review of brucellosis in the Middle East: disease frequency in ruminants and humans and risk factors for human infection. Epidemiol Infect 2016; 144(4):671-685.
    32. El-Hady AM, Sayed-Ahmed M, Saleh ME, et al. Seroprevalence and molecular epidemiology of brucellosis in cattle in Egypt. J Adv Dairy Res 2016; 4(2):153 doi: 10.4172/2329-888X.1000153.
    33. Xiao P, Yang H, Di D, et al. Genotyping of human Brucella melitensis biovar 3 isolated from Shanxi Province in China by MLVA16 and HOOF. PLoS One. 2015; 10(1):e0115932. doi: 10.1371/journal.pone. 0115932.34.
    34. Dastjerdi MZ, Nobari RF, Ramazanpour J. Epidemiological features of human brucellosis in central Iran, 2006–2011. Public Health 2012;126 (12):1058-1062.
    35. Mathew C, Stokstad M, Johansen T, et al. First isolation, identification, phenotypic and genotypic characterization of Brucella abortus biovar 3 from dairy cattle in Tanzania. BMC Vet Res 2015; 11(1):156. doi: 10.1186/s12917-015-0476-8
    36. Whatmore AM, Koylass MS, Muchowski J, et al. Extended multilocus sequence analysis to describe the global population structure of the genus Brucella: Phylogeography and relationship to biovars. Front Microbiol 2016; 7:2049. doi: 10.3389/fmicb.2016.02049.
    37. Ashrafganjooyi S, Saedadeli N, Alamian S, et al. Isolation and biotyping of Brucella spp. from sheep and goats raw milk in southeastern Iran. Trop Biomed 2017; 34(3):507-511.
    38. Jiang H, Fan M, Chen J, et al. MLVA genotyping of Chinese human Brucella melitensis biovar 1, 2 and 3 isolates. BMC Microbiol 2011; 11(1):256. doi: 10.1186/ 1471-2180-11-256.
    39. Gameel S, Mohamed S, Mustafa A, et al. Prevalence of camel brucellosis in Libya. Trop Anim Health Prod 1993; 25(2):91-93.
    40. Banai M, Mayer I, Cohen A. Isolation, identification, and characterization in Israel of Brucella melitensis biovar 1 atypical strains susceptible to dyes and penicillin, indicating the evolution of a new variant. J Clin Microbiol 1990; 28(5):1057-1059.
    41. Muendo EN, Mbatha PM, Macharia J, et al. Infection of cattle in Kenya with Brucella abortus biovar 3 and Brucella melitensis biovar 1 genotypes. Trop Anim Health Prod 2012; 44(1):17-20.
    42. Refai M. Incidence and control of brucellosis in the Near East region. Vet Microbiol 2002; 90(1-4):81-110.
    43. Ali S, Ali Q, Melzer F, et al. Isolation and identification of bovine Brucella isolates from Pakistan by biochemical tests and PCR. Trop Anim Health Prod 2014; 46(1):73-78.
    44. Al-Khalaf S, El-Khaladi A. Brucellosis of camels in Kuwait. Comp Immunol Microbiol Infect Dis 1989; 12(1-2):1-4.
    45. Erdenlig S, Sen A. Isolation and biotyping of Brucella species from aborted sheep fetuses. Pendik Vet Mikrobiyol Derg 2000; 31(2):31-42.
    46. Musa MT, Eisa MZ, El Sanousi EM, et al. Brucellosis in camels (Camelus dromedarius) in Darfur, Western Sudan. J Comp Pathol 2008; 138(2-3):151-155.
    47. Moghney FRA. A preliminary study on brucellosis on camels at Behira province. Ass Univ Bull Environ Res 2004; 7(1): 39-43.
    48. Abbas B, Agab H. A review of camel brucellosis. Prev Vet Med 2002; 55(1):47-56.
    49. Zaki R. Brucella infection among ewes, camels and pigs in Egypt. J Comp Pathol Ther 1948; 58(2):145-151.
    50. Ducrotoy M, Bertu W, Matope G, et al. Brucellosis in Sub-Saharan Africa: Current challenges for management, diagnosis and control. Acta Trop 2017; 165:179-193.
Veterinary Research Forum
Volume 10, Issue 4
Autumn 2019
Pages 315-321

  • Receive Date 11 July 2018
  • Revise Date 08 September 2018
  • Accept Date 25 September 2018

Home

Guide for Authors

Submit Manuscript

Reviewers

Contact Us