Examination of cross-reactivity among Trypanosoma evansi, Theileria annulata and  Babesia bigemina by Te-mAB-LAT and PCR

Reema Reema; S S Chaudhri; A Singh

doi:10.56093/ijans.v82i8.22996

Authors

Reema Reema Lala Lajpat Rai University of Veterinary and Animal Sciences, Uchani, Karnal, Haryana 132 001 India
S S Chaudhri Lala Lajpat Rai University of Veterinary and Animal Sciences, Uchani, Karnal, Haryana 132 001 India
A Singh Lala Lajpat Rai University of Veterinary and Animal Sciences, Uchani, Karnal, Haryana 132 001 India

https://doi.org/10.56093/ijans.v82i8.22996

Keywords:

Babesia bigemina, Cross-reactivity, Theileria annulata, Trypanosoma evansi, Monoclonal antibody-based antigen detection test, Polymerase chain reaction

Abstract

Blood and serum samples from 55 crossbred cows detected positive for haemoprotozoan infections (T. evansi-11, T. annulata- 23 and B. bigemina- 21) on examination of stained blood smears were collected and stored for T. evansi antigen detection monoclonal antibody- based latex agglutination test (Te-mAb-LAT) and polymerase chain reaction (PCR). Trypanosoma evansi mAb-LAT revealed circulating antigens in serum samples of 18 animals, including 11 confirmed positive for T. evansi, 3 confirmed positive for T. annulata and 4 confirmed positive for B. bigemina. The DNA fragments of the size 227 bp, 175 bp and 721 bp were amplified by PCR assays using specific primers for evansi, B. bigemina and T. annulata, respectively. The animals confirmed positive by stained smear examination for specific haemoprotozoa (T. evansi/ B. bigemina/ T. annulata) were also positive with specific PCR assay (T. evansi, Te- PCR/ B. bigemina, Bb-PCR/ T. annulata, Ta-PCR). The Te-PCR was observed free from any cross-reactivity with annulata/ B. bigemina whereas Ta-PCR revealed 5 of 21 B. bigemina infected samples positive for T. annulata and Bb-PCR detected 1 out of 11 T.evansi infected samples and 12 of 23 T. annulata infected samples positive for bigemina. It indicated that some of the animals had mixed infections of T. annulata and B. bigemina which is a common occurrence due to mixed infection of their vectors feeding on animals. Secondly, it was possible to have missed detection of low level of infection on microscopic examination of the stained blood smears.

The detection of circulating antigens in 7 blood samples (3 of T. annulata and 4 of B. bigemina infected animals) with Te-mAb-LAT, without their detection by Te-PCR was possibly due to previous exposure of these animals with evansi and subsequent treatment with trypanocidal drug. It is inferred that Te-mAb-LAT, being simple to perform, rapid, convenient and cost-effective could be quite suitable for diagnosis of trypanosomosis at field level and PCR being sensitive and specific has its utility at the laboratory level.

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References

D’Oliveira C, Weide MV, Habela M A, Jacquiet P and Jongejan F. 1995. Detection of Theileria annulata in blood samples of carrier cattle by PCR. Journal of Clinical Microbiology 33: 2665–69.

Figueroa J V, Chieves L P, Johnson G S and Buening G M. 1992. Detection of Babesia bigemina-infected carriers by polymerase chain reaction amplification. Journal of Clinical Microbiology 30: 2576–582.

García-Sanmartín J, Nagore D, García-Pérez A L, Juste R and Hurtado. 2006. Molecular diagnosis of Theileria and Babesia species infecting cattle in Northern Spain using reverse line blot macroarrays. BMC Veterinary Research 2: 16.

Ica A, Vatansever Z, Yildirim A, Duzlu O and Inci A. 2007. Detection of Theileria and Babesia species in ticks collected from cattle. Veterinary Parasitology 148: 156–60.

Kashiwazaki Y and Thammasart S. 1998. Effect of anti- immunoglobulin antibodies produced in cattle infected with Trypanosoma evansi on antigen detection ELISA. International Journal of Parasitology 28: 1353–60.

Lohr K F, Pholpark S, Siriwan P, Leesirikul N, Srikitjakarn L and Staak C. 1986. Trypanosoma evansi infection in buffaloes in north-east Thailand. 11. Abortions. Tropical Animal Health and Production 18: 103–08.

Mugittu K N, Silayo R S, Majiwa P A O, Kimbita E K, Mutayoba B M and Miscue R. 2001. Application of PCR and DNA probes in the characterization of trypanosomes in the blood of cattle in farms in Morogoro, Tanzania. Veterinary Parasitology 94: 177–89.

Pathak K M L and Singh N. 2005. Animal trypanosomosis. Intas Polivet 6: 194–99.

Rayulu V C, Singh A and Chaudhri S. S. 2007. Monoclonal antibody based immunoassays for detection of circulating antigens of Trypanosoma evansi in buffaloes. Italian Journal of Animal Sciences 6(8): 907–10.

Rayulu V C, Chaudhri S S and Singh A. 2009. Evaluation of parasitological and monoclonal antibody based assays in detection of Trypanosoma evansi infection in animals. Indian Journal of Animal Sciences 63(9): 978–81.

Sambrook J and Russell D W. 2001. Molecular Cloning: A Laboratory Manual. 3rd edn. Vol. 1 Cold Spring Harbour Lab Press. New York, USA.

Shyma, K.P. 2009. ‘Comparison of monoclonal antibody based latex agglutination test with PCR for diagnosis of Trypanosoma evansi in domestic animals.’ M V Sc. thesis, CCS Haryana Agricultural University, Hisar, India.

Singh A and Chaudhri S S. 2002. Comparison of efficiency of parasitological methods with Ag-ELISA in Trypanosoma evansi infected crossbred calves. Indian Journal of Animal Sciences 72 (1): 117–19.

Singh H, Mishra A K, Rao J R and Tewari A K. 2007. A PCR assay for detection of Babesia bigemina infection using clotted blood in bovines. Journal of Applied Animal Research 32: 201–02.

Thammasart S, Kanitpun R, Saithasao M and Kashiwazaki Y. 2001. Preliminary studies by ELISA on the antigen and antibody with Trypanosoma evansi in cattle. Tropical Animal Health and Production 33: 189–99.

Wernery U, Zachariah R, Mumford J A and Luckins T. 2001. Preliminary evaluation of diagnostic tests using horses experimentally infected with Trypanosoma evansi. Veterinary Journal 161: 287–300.

Woo P T K and Rogers D. 1974. A statistical study on the sensitivity of the haematocrit centrifugation technique in the detection of trypanosomes in blood. Transactions of Royal Society of Tropical Medicine and Hygiene 68: 319–26.

Wuyts N, Chodesajjawatee, N and Panyim S. 1994. A simplified and highly sensitive detection of Trypanosoma evansi by DNA amplification. Southeast Asian Journal of Tropical Medicine and Public Health 25: 266–71.