Prevalence of Brucella Melitensis and Brucella Abortus in Raw Milk and Dairy Product by Real Time PCR Technique

Year 2016, , 7 - 11, 01.01.2016

Yaran Majid Najafi Somayeh Shoaei Parisa Ataei Behrooz Fadaei Nobari Reza Ramazanpour Javad Farsi Mehdi Ahmadi Ahwaz Nasrin Khorasani Marzieh

https://doi.org/10.5455/umj.20151004054235

Abstract

Introduction: Brucellosis is a public health problem with a high incidence. The main causes of acquiring brucellosis are through the consumption of unpasteurized milk and dairy products. In this study we described a real-time PCR assay for more sensitive, rapid detection and differentiation of Brucella.abortus and Brucella.melitensis in raw milk and dairy products in Isfahan province. Materials and Methods: A Taq Man analysis and single step PCR BruAb2_0168BMEII0466 genes has performed in total of 132 unpasteurized milk samples and 65 dairy products. Samples were selected from various markets and animal farms located in different cities of Isfahan province. The duration period of sample collection was nearly two months May to June 2012 . Results: By real-time PCR 4 of 132 milk specimens 3% were positive for B. melitensis. Also one oil sample between 65 different dairy products 1.5% was positive for B. melitensis. None of the specimens were positive for B. abortus. We detected direct and incomplete correlation between brucella infected milk and different cities r=0.024, p value >0.05 . Conclusion: Results of present study indicated a relative high presence of this pathogen in raw milk. Real-time PCR is technically more simple, accurate, and rapid than current standard methods for identification of Brucella species .One of the most effective routes to control the disease includes pasteurization or boiling of milk and other dairy products for human consumption.

Keywords

Brucellosis, real time PCR, milk

References

• Baddour M M, Alkhalifa DH. Evaluation of 3 PCR techniques for detection of Brucella DNA in peripheral human blood. Can J Microbiol, 2008; 54(5):352-7.
• Seleem MN, Boyle SM. Brucellosis: a reemerging zoonosis. Vet Microbiol, 2010,27;140(3-4):392-8.
• Doosti A, Moshkelani S. The First Prevalence Report of Direct Identification and Differentiation of B.abortus and B. melitensis using Real Time PCR in House Mouse of Iran. WASET 2011,50:84-7
• Rijpens NP, Jannes G, Van Asbroeck M, Rossau R, Herman LM. Direct detection of Brucella spp. in raw milk by PCR and reverse hybridization with 16S-23S rRNA spacer probes. Appl Environ Microbiol. 1996; 62(5):1683-8.
• Young, EJ. Human Brucellosis. Rev Infect Dis1983;5:821-42.
• Arimi SM, Koroti E, Kang'ethe EK, Omore AO, McDermott JJ. Risk of infection with Brucella abortus and Escherichia coli O157:H7 associated with marketing of unpasteurized milk in Kenya. Acta Trop. 2005; 96(1):1-8.
• Minas M, Minas A, Gourgulianis K. Epidemiological and clinical aspects of human brucellosis in Central Greece. Jpn J Infect Dis. 2007;60(6):362-6.
• Doosti A, Ghasemi Dehkordi P. Application of REAL-TIME PCR for identification and differentiation of Brucella abortus and Brucella melitensisin cattle. Bulgar J Vet Med, 2011; (2): 109−115.
• Akbarmehr J. The prevalence of Brucella abortus and Brucella melitensis in local cheese produced in Sarab city, Iranand its public health implication. African J MR, 5(12), 1503, 18, 2011
• Pappas G, Papadimitriou P, Akritidis N, Christou L, Tsianos EV. The new global map of human brucellosis. Lancet Infect Dis, 2006; 6(2):91-9.
• Rabbani M, SharifiL. Ecological study of brucellosis in humans and animals in Khoy, a mountainous District of the IR of Iran. IJM, 2009;1(4):14-17.
• Refai M. Incidence and control of brucellosis in the Near East region. Vet Microbiol, 2002; 90(1-4):81-110.
• Maadi H, Moharamnejad M, Haghi M.Prevalence of Brucellosis in Cattle in Urmia, Iran. Iran Pak Vet J, 2010; (1): 81-82.
• Bertu WJ, Dapar M, Gusi AM, N gulukunS, Leo S. Prevalence of brucella antibodies in marketed milk in Jos and environs. Afr J Food Sci, 2010; 4(2): 62-64.
• Corbel MJ. Brucellosis: an overview. Emerg Infect Dis 1997; :213-21.
• Blasco JM. Control and eradication strategies for brucella melitensis infection in sheep and goats. Prilozi. 2010; (1):145-65.
• Leal-Klevezas DS, Martínez-Vázquez IO, López-Merino A, Martínez-Soriano JP. Single-step PCR for detection of Brucella spp. from blood and milk of infected animals. J Clin Microbiol. 1995; 33(12):3087-90.
• Hinić V, Brodard I, Thomann A, Cvetnić Z, Makaya PV, Frey J, Abril C. Novel identification and differentiation of Brucella melitensis, B. abortus, B. suis, B. ovis, B. canis, and B. neotomae suitable for both conventional and real-time PCR systems. J Microbiol Methods. 2008; 75(2):375-8.
• Shareef JM, 2006. A Review of serological investigations of brucellosis among farm animals and humans in Northern provinces of Iraq (1974-2004). J Vet Med B, 53: 38-40.
• Sofian M, Aghakhani A, Velayati AA, Banifazel M, Eslamifar A, Ramezani A. Risk factors for human brucellosis in Iran. Int J Infec Dis. 2008,12:157-161.