Comparison of diagnostic tests for the detection of bovine brucellosis in the natural cases of abortion

Document Type : Full paper (Original article)

Authors

1 Department of Animal Disease Research Centre, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana-141004, Punjab, India

2 Department of Veterinary Pathology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana-141004, Punjab, India

3 Department of Veterinary and Animal Husbandry Extension, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana-141004, Punjab, India

Abstract

Rapid and precise diagnosis in natural field cases of bovine abortion caused by Brucella abortus warrants the use of the most sensitive and reliable diagnostic methods. In the present study, bacterial isolation, serology, gross, histopathology, immunohistochemistry and polymerase chain reaction technique(s) were applied. Sero-prevalence studies showed the rate of 28.86% positive cases using the competitive ELISA. Histopathological changes were mainly seen in the placenta, fetal lungs, kidney, liver and spleen. Immunohistochemical (IHC) staining of Brucella spp. was evident as brown, finely granular intracytoplasmic staining in trophoblasts of placental sections and in section(s) of liver, lung, kidney and spleen. Twenty-eight out of the 103 samples (17 from stomach contents, 3 from placental cotyledons, 2 from vaginal discharges and 6 from pooled fetal tissues) produced 193 bp amplicon specific for Brucella genus. Moreover, the species-specific primers amplified a 498 bp amplicon which corresponded to B. abortus. Comparison of diagnostic tests revealed PCR and IHC provide a reliable test for the diagnosis of bovine brucellosis in aborted fetal tissue and placental cotyledons whereas serology is most important for detection of Brucella positive animals in a herd.

Keywords

References

Agrawal, R; Kumar, M and Singh, JL (2007). Sero-prevalence of brucellosis in Uttranchal. Indian Vet. J., 84: 204-205.
Alton, GG; Jones, LM and Pietz, DE (1975). Laboratory techniques in brucellosis. Monograph Series. World Health Organization, Geneva, Switzerland.
Anderson, ML (2007). Infectious causes of bovine abortion during mid to late gestation. Theriogenology. 68: 474-486.
Aulakh, HK; Patil, PK; Sharma, S; Kumar, H; Mahajan, V and Sandhu, KS (2008). A study on the epidemiology of bovine brucellosis in Punjab (India) using Milk-ELISA. Acta Vet. Brno., 77: 393-399.
Bricker, BJ (2002). PCR as a diagnostic tool for brucellosis. Vet. Microbiol., 90: 435-446.
Bricker, BJ and Halling, SM (1994). Differentiation of Brucella abortus bv. 1, 2 and 4, Brucella melitensis, Brucella ovis and Brucella suisbv 1 by PCR. J. Clin. Microbiol., 32: 2660-2666.
Cameron, A (1999). Survey Toolbox for Livestock Diseases -A Practical Manual and Software Package for Active Surveillance in Developing Countries. Australian Centre for International Agricultural Research Monograph. P: 330.
Chand, P and Sharma, AK (2004). Situation of brucellosis in bovines at organized cattle farms belonging to three different states. J. Immunol. Immunopathol., 6: 11-15.
Cortez, A; Scareelli, E; Soares, RM; Heinemann, MB; Sakamoto, SM; Genovez, ME; Ferreira, F and Richtzenhain, LJ (2001). Detection of Brucella DNA from aborted bovine fetuses by polymerase chain reaction. Aust. Vet. J., 79: 500-501.
Gall, D and Nielsen, K (2004). Serological diagnosis of bovine brucellosis: a review of test performance and cost comparison. Rev. Sci. Tech. Off. Int. Epiz., 23: 989-1002.
Geresu, MA and Kassa, GM (2016). A review on diagnostic methods of brucellosis. J. Veterinar. Sci. Techno., 7: 323.
Ilhan, Z; Aksakal, A; Ekin, IH; Gulhan, T; Solmaz, H and Erdenlig, S (2008). 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.
Keid, LB; Soares, RM; Vasconcellos, SA; Chiebao, DP; Megid, J; Salgado, VR and Richtzenhain, LJ (2007). A polymerase chain reaction for the detection of Brucella canis in semen of naturally infected dogs. Theriogenology. 67: 1203-1210.
Leal-Klevezas, DS; Martínez-Vázquez, IO; García-Cantu, J; López-Merino, A and Martínez-Soriano, JP (1995). Single-step PCR for detection of Brucella spp. from blood and milk of infected animals. J. Clin. Microbiol., 33: 3087-3090.
Leuenberger, R; Boujon, P; Thür, B; Miserez, R; Garin-Bastuji, B; Rüfenacht, J and Stärk, KD (2007). Prevalence of classical swine fever, Aujeszky’s disease and brucellosis in a population of wild boar in Switzerland. Vet. Rec., 160: 362-368.
López, A; Hitos, F; Perez, A and Navarro-Fierro, RR (1984). Lung lesions in bovine fetuses aborted by Brucella abortus. Can. J. Comp. Med., 48: 275-277.
Luna, LG (1968). Manual of histologic staining methods of the Armed Forces Institute of Pathology. 3rd Edn., New York, McGraw-Hill. PP: 32-39.
Meador, VP and Deyoe, BL (1989). Intracellular localization of Brucella abortus in bovine placenta. Vet. Pathol., 26: 513-515.
Meador, VP; Deyoe, BL and Cheville, NF (1989). Pathogenesis of Brucella abortus infection of the mammary gland and supramammary lymph node of the goat. Vet. Pathol., 26: 357-368.
Meador, VP; Tabatabai, LB; Hagemoser, WA and Deyoe, BL (1986). Identification of Brucella abortus in formalin fixed, paraffin embedded tissues of cows, goats, and mice with an avidin biotin peroxidase complex immuno-enzymatic staining technique. Am. J. Vet. Res., 47: 2147-2150.
Muñoz, PM; Marín, CM; Monreal, D; González, D; Garin-Bastuji, B; Díaz, R; Mainar-Jaime, RC; Moriyón, I and Blasco, JM (2005). Efficacy of several serological tests and antigens for diagnosis of bovine brucellosis in the presence of false positive serological results due to Yersinia enterocolitica O: 9. Clin. Diagn. Lab. Immunol., 12: 141-151.
Nielsen, K (2002). Diagnosis of brucellosis by serology. Vet. Microbiol., 90: 447-459.
Paweska, JT; Potts, AD; Harris, HJ; Smith, SJ; Viljoen, GJ; Dungu, B; Brett, OL; Bubb, M and Prozesky, L (2002). Validation of an indirect enzyme-linked immunosorbent assay for the detection of antibody against Brucella abortus in cattle sera using an automated ELISA workstation. Onderstepoort. J. Vet. Res., 69: 61-77.
Pérez, J; Quezada, M; Lopez, J; Casquet, O; Sierra, MA and Martin De Las Mulas, J (1998). Immuno-histochemical detection of Brucella abortus antigens in tissues from aborted bovine fetuses using commercially available polyclonal antibody. J. Vet. Diagn. Invest., 10: 17-21.
Poester, FP; Samartino, LE and Lage, AP (2005). Diagnóstico da brucelose bovina. Cad. Téc. Vet. Zootec., 47: 13-29.
Rahman, MS; Faruk, MO; Her, M; Kim, JY; Kang, SI and Jung, SC (2011). Prevalence of brucellosis in ruminants in Bangladesh. Vet. Med. Czech., 56: 379-385.
Rahman, MA; Islam, MS; Alam, MGS and Shamsuddin, M (1997). Seroprevalence of Brucellosis in the buffalo of a selected area in Bangladesh. Buff. J., 2: 209-214.
Richtzenhain, LJ; Cortez, A; Heinemann, MB; Soares, RM; Sakamoto, SM; Vasconcellos, SA; Higa, ZM; Scarcelli, E and Genovez, ME (2002). A multiplex PCR for the detection of Brucella spp. and Leptospira spp. DNA from aborted bovine fetuses. Vet. Microbiol., 87: 139-147.
Samartino, LE and Enright, FM (1993). Pathogenesis of abortion of bovine brucellosis. Comp. Immunol. Microbiol. Infect. Dis., 16: 95-101.
Sandhu, KS; Filia, G; Sharma, DR; Dhand, NK; Singh, J and Saini, SS (2001). Prevalence of brucellosis among dairy animals of Punjab. Indian J. Comp. Microbiol. Immunol. Infect. Dis., 22: 160-161.
Sharma, S; Alka; Mahajan, V; Verma, S; Kaur, K; Meenakshi and Kumar, H (2007). Screening of dairy farms of Punjab (India) for brucellosis and para-tuberculosis. Indian Vet. J., 84: 315-316.
Singh, G; Sharma, DR; Sandhu, KS and Dhand, NK (2002). Economic losses occurring due to bovine abortions in Punjab. In: 10th International Congress of Asian-Australasian Association of Animal Production Societies. 23-27 September. Ashoka Hotel, New Delhi. Indian Association of Animal Production and World Buffalo Trust, New Delhi, India.
Surucuoglu, S; Sibel, El; Ural, S; Gazi, H; Kurutepe, S; Taskiran, P and Yurtsever, SG (2009). Evaluation of real-time PCR method for rapid diagnosis of brucellosis with different clinical manifestations. Pol. J. Microbiol., 58: 15-19.
Zamri-Saad, M and Kamarudin, MI (2016). Control of animal brucellosis: the Malaysian experience. Asian Pac. J. Trop. Med., 9: 1136-1140.

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