Indian Journal of Animal Research

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Indian Journal of Animal Research, volume 52 issue 4 (april 2018) : 591-598

Immunodiagnostic potency of homologous antigens for natural Haemonchus contortus infection in small ruminants in plate and paper enzyme linked immunosorbent assay

Bhupamani Das, Niranjan Kumar, Mehul M. Jadav, Jayesh B. Solanki
1<p>Department of Parasitology, Vanbandhu College of Veterinary Science and Animal Husbandry,&nbsp;Navsari Agricultural University, Navsari-396 450, Gujarat, India.</p>
Cite article:- Das Bhupamani, Kumar Niranjan, Jadav M. Mehul, Solanki B. Jayesh (2016). Immunodiagnostic potency of homologous antigens for natural Haemonchus contortus infection in small ruminants in plate and paperenzyme linked immunosorbent assay . Indian Journal of Animal Research. 52(4): 591-598. doi: 10.18805/ijar.v0i0f.3789.

The objective of this study was to characterize Haemonchus contortus antigens, and to standardize and evaluate indirect plate and dot-ELISA using homologous antigens in the small ruminants. Electrophoretic separation of somatic antigen in reducing condition on 15% polyacrylamide gel resolved into 16 proteins of molecular weight ranging from 14-100 kDa. Two step ethanolic extraction of the supernatant of in-vitro culture of H. contortus yielded excretory-secretory (E-S) antigen/ cathepsin L cysteine proteinase of molecular weight 28 kDa. The animals (Goats=103; Sheep=91) were broadly kept into post-mortem (PM) and faecal examined groups and further sub-grouped based on mono or multiply helminths infection. At many occasion, the somatic antigen found to cross reacts with other helminths parasites thus minimizing the specificity of the selected tests and antigens. There was no any direct correlation between the parasites load and ELISA reactivity pattern. The prevalence rate of haemonchosis was 55.7 (34/61) in goats/ 47.6 (40/84) % in sheep as per PM examination while it was 45.63 (47/103) in goats/ 41.76% (38/91) in sheep and 36.89 (38/103) in goats/ 35.16% (32/91) in sheep using E-S antigen based plate and dot-ELISA, respectively. With E-S antigen, the overall % sensitivity, specificity, positive and negative predictive values of plate-ELISA was 89.74 (goats)/ 80.95 (sheep), 81.25 (goats)/ 91.84 (sheep), 74.47 (goats)/ 89.47 (sheep), 92.86 (goats)/ 84.91 (sheep), respectively while for dot-ELISA it was 66.67 (goats)/ 61.9 (sheep), 81.25 (goats)/ 87.76 (sheep), 68.42 (goats)/ 81.25 (sheep), 80 (goats)/ 72.88 (sheep), respectively, so the tests and E-S antigen can be recommended for the detection haemonchosis in the small ruminants.  

  1. Abramatov, M.B.; Amirov, O.O.; Kuchboev, A.E.; Khalilov, I.M.; Abdurakhmanov. I.Y. (2013). Morphological and molecular characterization of Haemonchus contortus and H. placei (Nematoda: Trichostrongylidae) from Uzbekistan by sequences of the second internal transcribed spacer of ribosomal DNA. Sci. Parasitol. 14: 115-120. 

  2. Allaie, I.M.; Prasad, A. and Nasir, A. (2010). Identification of immunodominant polypeptides of infective larva of Haemonchus contortus. Indian J. Ani. Sci. 80: 506-508.

  3. Arunkumar, S. and Sangaran, A. (2009). An immunological analysis on sharing of antigens amongst gastro-intestinal nematodes using inhibition ELISA. Indian J. Sci. Technol. 2: 33-35.

  4. Arunkumar, S.; Basith, S.A. and Gomathinayagam, S. (2011). Serum antibody responses of madras red sheep to excretory antigen of Haemonchus contortus. Int. J. Agro Vet. Med. Scie. 5: 523-529.

  5. Arunkumar, S.; Basith, S.A. and Gomathinayagam, S. (2012). A comparative analysis on serum antibody levels of sheep immunized with crude and thiol-purified excretory/secretory antigen of Haemonchus contortus. Vet. World 5: 279-284.

  6. Coles, G.C. and Rubano, D. (1988). Antigenicity of a proteolytic enzyme of Fasciola hepatica. J. Helminthol. 62: 257-    260.

  7. Dixit, A.K.; Yadav, S.C. and Sharma, R.L. (2002). The 28 kDa Fasciola gigantica cysteine proteinase in the diagnosis of prepatent ovine fasciolosis. Vet. Parasitol. 109: 233-247.

  8. El Hassan, E.M. and El Bahr, S.M. (2012). Antigenic and immunogenic components of Haemonchus longistipes identified by western immunobloting. Am. J. Biochem. Biotechnol. 8: 164-170.

  9. Fagbemi, B.O. and Guobadia, E.E. (1995). Immunodiagnosis of fasciolosis in ruminnats using a 28-kDa cysteine protease of Fasciola gigantica adult worms. Vet. Parasitol. 57: 309-318.

  10. Go´mez-Mun˜oz, M. T.; Domnýguez, I.A.; Gomez-Iglesias, L.A.; Fernandez-Perez, F.; Me´ndez, S.; de la Fuente, C.; Alunda, J.M. and Cuquerella, M. (2000). Serodiagnosis of haemonchosis with a somatic antigen (Hc26) in several breeds of sheep. J. Vet. Dia. Inv. 12: 354-360. 

  11. Gowda, A.K.J. (2016a). Sero-prevalence of Haemonchus contortus infection in sheep by Indirect ELISA using somatic antigen. J. Parasit. Dis. 40: 464-468.

  12. Gowda, A.K.J. (2016b). Immunodiagnosis of Haemonchus contortus infection in sheep by counter immuno-electrophoresis using somatic antigen. J. Parasit. Dis. doi: 10.1007/s1263901405272.

  13. Gupta S.K. and Singla L.D. (2012) Diagnostic trends in parasitic diseases of animals. In: Veterinary Diagnostics: Current Trends. Gupta RP, Garg SR, Nehra V and Lather D (Eds), Satish Serial Publishing House, Delhi, pp 81-112.

  14. Jas, R.; Ghosh, J.D. and Das, K. (2010). Polyclonal antibody based coproantigen detection immunoassay for diagnosis of Oesophagostomum columbianum infection in goats. Vet. Parasitol. 170: 262-267.

  15. Kaur, K.; Kapur, J.; Parmar, A. and Sood, M.L. (2002). Identification of immunodominant antigens of adult Haemonchus contortus (Nematoda: Trichostrongylidae). J. Vet. Med. B 49: 260-262.

  16. Kumar, N.; Ghosh, S. and Gupta, S.C. (2008a). Detection of Fasciola gigantica infection in buffaloes by enzyme-linked immunosorbent assay. Parasitol. Res. 104: 155-161. 

  17. Kumar, N., Ghosh, S. and Gupta, S.C. (2008b). Early detection of Fasciola gigantica infection in buffaloes by enzyme-    linked immunosorbent assay and dot enzyme-linked immunosorbent assay. Parasitol. Res. 103: 141-150. 

  18. Laemmli, U.K. (1970). Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227: 630-    684.

  19. Lone, B.A.; Chishti, M.Z.; Ahmad, F.; Tak, H. and Hassan, J. (2012) Immunodiagnosis of Haemonchus contortus infection in sheep by indirect enzyme linked immunosorbent assay (ELISA). Iran. J. Vet. Res. 13: 49-53.

  20. Mir, R.A.; Chishti, M.Z.; Zargar, M.A.; Hidayatullah T. and Ganie, S.A. (2008). Excretory-secretory antigens are better than crude antigens for serodiagnosis of Haemonchus contortus. Asian J. Sci. Res. 1: 171-175. 

  21. Miranda-Miranda, E.; Murillo-Sánchez, M.H. and Cossío-Bayúgar, R. (2007). Use of the Baculovirus heterologous gene system expression on the generation of a hybrid recombinant antigen useful in the immunodiagnostics of ruminant hemonchosis. Bioquimia 32: 83-90.

  22. 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 leishmaniosis. J. Immunol. Methods 64: 205-214.

  23. Prasad, A.; Nasir, A. and Singh, N. (2007). Immunodominant polypeptides in somatic antigen of adult Haemonchus contortus by western blotting and immunoprecipitation. Indian J. Ani. Sci. 77: 533-537.

  24. Prasad, A.; Nasir, A. and Singh, N. (2008). Detection of anti-Haemonchus contortus antibodies in sheep by dot-ELISA with immunoaffinity purified fraction of E-S antigen during prepatency. Indian J. Exp. Biol. 46: 94-99. 

  25. Rathore, D.K.; Suchitra, S.; Saini, M.; Singh, B.P. and Joshi, P. (2006). Identification of a 66 kDa Haemonchus contortus excretory/secretory antigen that inhibits host monocytes. Vet. Parasitol. 138: 291-300.

  26. Sackett, D.; Holmes, P.H.; Abbott, K.; Jephcott, S. and Barber, M. (2006). Assessing the economic cost of endemic disease on the profitability of Australian beef cattle and sheep producers, AHW.087 Report, Meat and Livestock Australia, North Sydney, 119.

  27. Singla, L.D. (2011). Synthetic peptides as immunodiagnostic agents of parasitic diseases. In: Compendium of Winter School on Advanced Molecular Biology Tools Used in Animal Disease Diagnosis and Development of New Generation Vaccines held from October 03-23, 2011 at GADVASU, Ludhiana, pp 273-283.

  28. 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.

  29. Tysker, M.; Ploeger, H.W.; Martin, R.J.; Schallig, H.D. and Chappel, L.H. (2000). Value of present diagnostic methods for gastrointestinal nematode infections in ruminants. Symposia of the British Society for Parasitology, Volume 37. Vet. Parasitol. - recent developments in immunology, epidemiology and control. 120: supplement.


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