Genomic detection of Brucella spp in Seropositive cattle in charmahal va Bakhtiyari province, Iran

Authors

1 Department of Pathobiology, Faculty of Veterinary Medicine, University of Shahrekord, Shahrekord-Iran

2 Veterinary directorate general of Chaharmahal and Bakhtiyari, Shahrekord- Iran

3 Graduated from the Faculty of Veterinary Medicine, Shahrekord University, Shahrekord- Iran

4 Reasearch Institute of Zoonotic Diseases, Shahrekord University

Abstract

BACKGROUND: Brucellosis is one of the most common zoonosis in Middle East and Iran. OBJECTIVES: The purpose of this study was genomic detection of Brucella spp. in sero-positive dairy cattle. METHODS: We have collected 28,519 blood samples from cows during 2012-2013. Samples were screened by Slide and tube agglutination and 2-Mercaptoethanol tests. Samples with anti-Brucella antibodies titer ≥ 1:80 and ≥1:40 in tube agglutination and 2-ME tests were considered as positive respectively. Tissue samples include: lymph nodes, liver, testicle and kidney from 122 samples of slaughtered cows were collected. The Sero-positive samples were examined by a collection of specific primers for Brucella abortus, Brucella melitensis, vaccinal strains included RB51 and Rev1 using PCR tests. RESULTS: Results showed that 450 samples were positive in slide agglutination test and 447 samples had anti-brucella antibodies titer equal to or more than 1:80. So they were positive by tube agglutination test. Three hundred eighty nine samples were positive by 2- mercaptoethanol test. PCR test results showed that 46 samples (37.7%) out of 122 samples had a specific sequence of Brucella or otherwise they have an active infection with Brucella species, whereas 62.3% of samples were negative. The PCR results showed that 2 samples (4.35%) were infected by B. melitensis, 2 samples (4.35%) infected by Rev1 strain and 42 samples (91.3%) were infected by B. abortus. CONCLUSIONS: The results showed that, as we had expected, the majority of cows were infected by B. abortus. Animals who infected by B. melitensis and Rev1 strain may be a result of contact with sheep or goats. We couldn’t find Brucella genome in 76 samples (62.3%) of sero-positive cows. It may be caused by cross reaction of sera with Brucella species in tests or activation of immune system response and elimination of organism from internal organs.

Keywords

References

Al-Jam’a, AH., Elbashir, A.M., Al-Faris, S.S. (1993) Brucella pneumonia: A case report. Ann Saudi Med. 13: 74-77.
Budras, K.D.,  McCarthy Al Dahouk, S., Nockler, K., Scholz, H.C., Pfeffer, M., Neubauer, H., Tomaso, H. (2007) Evaluation of genus-specific and species-specific real- time PCR assays for the identification of Brucella spp. Clin Chem Lab Med. 45: 1464-1470.
Bogdanovich, T., Skurnik, M., Lubeclek, P.S., Ahrens, P., Hoorfar, J. (2004) Validated 5’ nuclease PCR assay for rapid identification of the genus Brucella. J Clin Microbiol. 42: 2261-2263.
Bounaadja, L., Albert, D., Chenais, B., Henault, S., Zygmunt, M., Poliak, S., Garin-Bastuji, B.  (2009) Real-time PCR for identification of Brucella spp: A comparative study of Is711, bcsp31 and per target genes. Vet Microbiol. 137: 156-164.
Cloeckaert, A., Grayon, M., Grepinet, O. (2000) An IS711 element downstream of the bp 26 gene is a specific of Brucella spp. isolated from marine mammals. Clin Diagn Lab Immunol. 7: 835-839.
Cloeckaert, A., Grayon, M., Grepinet, O. (2002)Identification of Brucella melitensis vaccine strain Rev.1 by PCR-RFLP based on a mutation in the rpsL gene. Vaccine. 20: 2546–2550.
Esmaili, H., Tajic, P., Ekhtiarzadeh, H., Bolorchi, M., Hamedi, M., Khalaj, M., Amiri, K. (2012) Evaluation of bovine brucellosis control and eradication program in Iran: An epidemiological survey. J Vet Res (In Persian). 67: 211-221.
Ghosian Moghaddam, M.H., Keyvani Amineh, H., Zahraei Salehi, T., Khazraei Nia, P., Fallah, N. (2008) A comparison of culture and PCR methods with wrights test for diagnosis of Brucellosis. J Shahed University (In Persian). 74: 51-58.
Gross, A., Bertholet, S., Mauel, J., Dornand, J. (2004) Impairment of Brucella growth in human macrophagic cells that produce nitric oxide. Microb Pathog. 36: 75-82.
Ghorbani, A., Rabbani Khorasgani, M., Zarkesh Esfahani, H., Sharifiyazdi, H., Dehghan Kashani, A., Emami, H. (2013) Comparison of serology, culture, and PCR for detection of brucellosis in slaughtered camels in Iran. Comp Clin Pathol. 22: 913- 917.
Ilhan, Z., Aksakal, A., Ekin, I.H., Gulhan, T., Solmaz, H., Erdenlig, S. (2007) Comparison of culture and PCR for the detection of Brucella melitensis in blood and lymphoid tissues of serologically positive and negative slaughtered sheep. Lett Appl Microbiol. 46: 301-6.
Leboffe, M.J., Pierce, B.E. (2012) A Photographic Atlas for the Microbiology Laboratory. (4th ed.) Morton Publishing Company. Englewood, Colorado. p. 138.
Lihan, Z., Aksakal, A., Ekin, IH., Guihan, A., Solmaz, H., Erdenlig, S. (2007) Comparison of culture and PCR for the detection of Brucella melitensis in blood and lymphoid tissues of serologically positive and negative slaughtered sheep. Lett Appl Microbiol. 46: 301-306.
López-Goñi, I., García-Yoldi, D., Marín, CM., de Miguel, MJ., Muñoz, PM., Blasco, JM., Jacques, I., Grayon, M., Cloeckaert, A., Ferreira, A.C., Cardoso, R., Correa de Sa, M.I., Walravens, K., Albert, D., Garin-Bastuji, B. (2008) Evaluation of a multiplex PCR assay (Bruce-ladder) for molecular typing of all Brucella species, including the vaccine strains. J Clin Microbiol. 46: 3484–3487.
Lübeck, PS., Skurnik, M., Ahrens, P., Hoorfar, J. (2003) A multiplex PCR detection assay for Yersinia enterocolitica serotype 0:9 and Brucella spp. based on the perosamine synthetase gene. Application to Brurella diagnostics. Adv Exp Med Biol. 529: 451-453.
Mayer-Scholl, A1., Draeger, A., Göllner, C., Scholz, HC., Nöckler, K. (2010) Advancement of a multiplex PCR for the differentiation of all currently described Brucella species. J Microbiol Meth. 80: 112–114.
Mostafavi, A., Asmand, M. (2012) Process of brucellosis (Malta fever) in Iran during 1993-2006. Iranian Journal of Epidemiology (In Persian). 8: 94-101.
Nielsen, K., Smith, P., Widdison, J., Gall, D., Kelly, L., Kelly, W., Nicoletti, P. (2004) Serological relationship between cattle exposed to Brucella abortus, Yersinia enterocolitica O:9 and Escherichia coli O157: H7. Vet Microbiol. 100: 25-30.
 Pishva, E., Salehi, M. (2008) First report of isolation of Brucella melitensis, vaccine strain Rev.1 as a source of cattle infection in Iran. Journal of Sciences, Islamic Republic of Iran. 19: 19-23.
Quinn, PJ., Carter, ME., Markey, B., Carter, GR. (2004) Clinical Veterinary Microbiology. (1st ed.) Morton Publishing Company. Philadelphia, USA.
Rossetti, CA., Galindo, CL., Garner, HR., Adams, LG. (2011) Transcriptional profile of the intracellular pathogen Brucella melitensis following HeLa cells infection. Microb Pathogen. 51: 338–344.
Schmoock, G., Ehrichtb, R., Melzera, F., Elschnera, M., Tomasoa, H., Neubauera, H., Al Dahouk, S. (2011) Development of a diagnostic multiplex polymerase chain reaction microarray assay to detect and differentiate Brucella spp. Diagn Microb Infect Dis. 71: 341–353.
Scholz, HC., Hubalek, Z., Sedlácek, I., Vergnaud, G., Tomaso, H., Al Dahouk, S., Melzer, F., Kampfer, P., Neubauer, H., Cloeckaert, A., Maquart, M., Zygmunt, M.S., Whatmore, A.M, Falsen, E., Bahn. P., Gollner, C., Pfeffer, M., Huber, B., Busse, H.J., Nockler, K. (2008) Brucella microti sp. nov., isolated from the common vole Microtus arvalis. Int J Syst Evol Microbiol. 58: 375-382.
Zeinalian Dastjerdi, M., Fadaei Nobari, R., Ramazanpour, J. (2012) Epidemiological features of human brucellosis in central Iran, 2006–2011. Public health. 126: 1058-1062.
Zeinalian, M., Shirzadi, MR., Kianpour, M. (2012) National guideline for Brucellosis control. In: Bovine Brucellosis Control. Ministry of health and medical education. Health deputy, Center of communicable disease control, Zoonoses office. (2nd ed) Razeh Nahan, Tehran, Iran. p. 40-44.  
Journal of Veterinary Research
Volume 70, Issue 4 - Serial Number 4
December 2015
Pages 395-401

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