Article history: Received: 23-06-2017, Accepted: 11-09-2017, Published online: 06-10-2017
Corresponding author: Niranjan Kumar
E-mail: firstname.lastname@example.orgCitation: Kumar N, Varghese A, Solanki JB (2017) Seroprevalence of Fasciola gigantica infection in bovines using cysteine proteinase dot enzyme-linked immunosorbent assay, Veterinary World, 10(10): 1189-1193.
Aim: The objective of the present study was to know the seroprevalence status of Fasciola gigantica infection in cattle and buffaloes using cysteine proteinase (CP) antigen in dot enzyme-linked immunosorbent assay (ELISA) format under field conditions.
Materials and Methods: As per the standard protocol, the sera were collected from the blood of 112 cattle and 38 buffaloes of coastal areas of Navsari district, South Gujarat, India. The indirect ELISA was performed on the strip of nitrocellulose paper blotted with 1 μl of CP antigen, to detect F. gigantica seropositive animals.
Results: The native CP of F. gigantica revealed a single visible band on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. There was no any noted cross-reaction between the selected antigen and sera of Gastrothylax crumenifer-infected animals in ELISA. Out of 150 screened bovines, the sera of 47 (31.33%) were found to be reactive in dot-ELISA, with a prevalence rate of 31.25% and 31.58% in cattle and buffaloes, respectively. The seropositive bovines with heavy, moderate, and light level of infection were 44.68%, 34.04%, and 21.28%, respectively (p<0.05 between heavy and light; p>0.05 between moderate and heavy or light). The share of F. gigantica seropositive and negative animals was 31% and 69%, respectively. The optical density at 450 nm of pooled sera of seropositive bovines with heavy, moderate, and light reactivity in plate-ELISA was significantly higher with field or reference negative sera.
Conclusion: The CP-based dot-ELISA can be useful for field veterinarians for quick and timely isolation of the animals requiring urgent flukicide therapy.
Keywords: bovines, cysteine proteinase, dot enzyme-linked immunosorbent assay, Fasciola gigantica.
1. Gupta, S.C., Ghosh, S., Raina, O.K., Joseph, D., Preeti, R., Singh, B.P., Mishra, A.K., Chandra, D. and Samanta, S. (2008) Status and prevalence of fasciolosis in cattle and buffaloes in different agro-climatic zones of Uttar Pradesh. J. Vet. Parasitol., 22: 59-63.
2. Pandya, S.S., Hasnani, J.J., Patel, P.V., Chauhan, V.D., Hirani, N.D., Shukla, R. and Dhamsaniya, H.B. (2015) Study on prevalence of fasciolosis in buffaloes at Anand and Ahmedabad districts, Gujarat, India. Vet. World, 8(7): 870-874. [Crossref] [PubMed] [PMC]
3. Elelu, N., Ambali, A., Coles, G.C. and Eisler, M.C. (2016) Cross-sectional study of Fasciola gigantica and other trematode infections of cattle in Edu local government area, Kwara state, North-Central Nigeria. Parasit. Vectors, 9: 470. [Crossref] [PubMed] [PMC]
5. WAAVP Congress. (2005) Fasciola hepatica: Suppression of Host Immune Responses and Susceptibility of Other Diseases, Annual Convention of the World Association for the Advancement of Veterinary Parasitology. p21-22.
7. Jean-Richard, V., Crump, L., Abicho, A.A., Nare, N.B., Greter, H., Hattendorf, J., Schelling, E. and Zinsstag, J. (2014) Prevalence of Fasciola gigantica infection in slaughtered animals in south-eastern Lake Chad area in relation to husbandry practices and seasonal water levels. BMC Vet. Res., 10: 81. [Crossref]
9. Gupta, S.C. and Yadav, S.C. (1992) Sexual maturity of Fasciola gigantica in experimentally infected rabbits, goats and buffaloes. Indian J. Parasitol., 16: 133-134.
10. Rehman, T., Khan, M.N., Abbas, R.Z., Babar, W., Sikandar, A. and Zaman, M.A. (2016) Serological and coprological analyses for the diagnosis of Fasciola gigantica infections in bovine hosts from Sargodha, Pakistan. J. Helminthol., 90(4): 494-502. [Crossref] [PubMed]
11. Valero, M.A., Bargues, M.D., Khoubbane, M., Artigas, P., Quesada, C., Berinde, L., Ubeira, F.M., Mezo, M., Hernandez, J.L., Agramunt, V.H. and Mas-Coma, S. (2016) Higher physiopathogenicity by Fasciola gigantica than by the genetically close F. hepatica: Experimental long-term follow-up of biochemical markers. Trans R. Soc. Trop. Med. Hyg., 110(1): 55-66. [Crossref] [PubMed]
12. Varghese, A., Raina, O.K., Nagar, G., Garg, R., Banerjee, P.S., Maharana, B.R. and Kollannur, J.D. (2012) Development of cathepsin-L cysteine proteinase based dot-enzyme-linked immunosorbent assay for the diagnosis of Fasciola gigantica infection in buffaloes. Vet. Parasitol., 183: 382-385. [Crossref]
13. Yokananth, S., Ghosh, S., Gupta, S.C., Suresh, M.G. and Saravanan, D. (2005) Characterization of specific and cross-reacting antigens of Fasciola gigantica by immunoblotting. Parasitol. Res., 97: 41-48. [Crossref] [PubMed]
15. Kumar, N., Ghosh, S. and Gupta, S.C. (2008) Early detection of Fasciola gigantica infection in buffaloes by enzyme-linked immunosorbent assay and dot enzyme-linked immunosorbent assay. Parasitol. Res., 103: 141-150. [Crossref]
16. Yadav, S.C., Saina, M., Raina, O.K., Nambi, P.A., Jadav, K. and Sriveny, D. (2005) Fasciola gigantica cathepsin-L cysteine proteinase in the detection of early experimental fasciolosis in ruminants. Parasitol. Res., 97: 527-534. [Crossref]
17. Sriveny, D., Raina, O.K., Yadav, S.C., Chandra, D., Jayraw, A.K., Singh, M., Velusamy, R. and Singh, B.P. (2006) Cathepsin-L cysteine proteinase in the diagnosis of bovine Fasciola gigantica infection. Vet. Parasitol., 135: 25-31. [Crossref] [PubMed]
18. Avcioglu, H., Guven, E., Balkaya, I., Kaynar, O. and Hayirli, A. (2014) Evaluation of coprological and serological techniques for diagnosis of bovine fasciolosis. Isr. J. Vet. Med., 69(4): 203-210.
19. Yadav, P. and Singh, R. (2011) A review on anthelmintic drugs and their future scope. Int. J. Pharm. Pharm. Sci., 3(3): 17-21.
20. Raina, O.K., Yadav, S.C., Sriveny, D. and Gupta, S.C. (2006) Immuno-diagnosis of bubaline fasciolosis with Fasciola gigantica cathepsin-L and recombinant cathepsin L 1-D protease. Acta Trop., 98: 145-151. [Crossref] [PubMed]
21. Ouchterlony, O. (1964) Gel diffusion technique. In: Ackroyd, J.F., editor. Immunological Methods. Blackwell Scientific Publication, Oxford. p55-78.
23. Pappas, M.G., Hajkowski, R. and Hockmeyer, W.T. (1983) Dot-enzyme linked immunosorbent assay (dot-ELISA): A micro technique for the rapid diagnosis of visceral leishmaniasis. J. Immunol. Methods, 64: 205-214. [Crossref]
24. Maisonnave, J. (1999) Standardization of dot immuno-peroxidase assay for field diagnosis of Fasciola hepatica infected cattle. Vet. Parasitol., 85: 259-268. [Crossref]
25. Cornelissen, J.B.W., Gassenbeek, C.P.H., Borgsteede, F.H.M., Holland, W.G., Harmsen, M.M. and Bocrsma, W.J.A. (2001) Early immune-diagnosis of fasciolosis in ruminants using recombinant Fasciola hepatica cathepsin-L-like protease. Int. J. Parasitol., 31: 728-737. [Crossref]
26. Fagbemi, B.O. and Guobadia, E.E. (1995) Immuno-diagnosis of fasciolosis in ruminants using a 28 kDa cysteine protease of Fasciola gigantica adult worms. Vet. Parasitol., 57: 309-318. [Crossref]
27. Damwesh, S.D. and Ardo, M.B. (2013) Detection of Fasciola gigantica antibodies using pourquier ELISA kit. Sokoto J. Vet. Sci., 11(2): 43-48. [Crossref]
28. Chamuah, J.K., Jacob, S.S., Sakhrie, A. and Borkotoky, D. (2014) Serological prevalence of Fasciola gigantica in mithun (Bos frontalis). Int. J. Livest. Res., 4(5): 15-20. [Crossref]
29. Sabapara, G.P., Desai, P.M., Kharadi, V.B., Saiyed, L.H. and Singh, R.R. (2010) Housing and feeding management practices of dairy animals in the tribal area of South Gujarat. Indian J. Anim. Sci., 80: 1022-1027.
30. Palmer, D.G., Lyon, J., Palmer, M.A. and Forshaw, D. (2014) Evaluation of a copro-antigen ELISA to detect Fasciola hepatica infection in sheep, cattle and horses. Aust. Vet. J., 92(9): 357-361. [Crossref] [PubMed]31. Abdolahi-Khabisi, S. and Sarkari, B. (2016) Detection of Fasciola hepatica and Fasciola gigantica common and uncommon antigens, using rabbit hyper immune serum raised against their excretory-secretory and somatic antigens. J. Parasit. Dis., 40(4): 1552-1557. [Crossref] [PubMed]