Diagnostic potential of 36-55 kDa somatic antigens of Fasciola gigantica for bovine fasciolosis
DOI:
https://doi.org/10.56825/bufbu.2022.4113170Keywords:
Bubalus bualis, buffaloes, fascioliasis, ruminants, antigens, western blotting, ELISA, SDS-PAGE, PakistanAbstract
Fascioliasis causes huge economic losses in livestock industry by reducing the growth, fertility rate, meat and milk yield. In the present study, somatic antigens isolated from Fasciola gigantica were assessed for the early detection of infection by developing antibody detection enzyme immunoassay. The somatic antigens extracted from adult F. gigantica worms was separated on SDS-PAGE and immunogenicity was determined by Western blot (WB). The results showed polypeptides bands between 15 to 95 kDa, and most prominent bands were 15, 28, 36, 38, 55, 72 and 95 kDa polypeptides. The WB revealed a single polypeptide of size between 36 to 55 kDa as antigenic. The sensitivity and specificity of ELISA test established for 36 to 55 kDa somatic antigens was 95.45% (95% CI: 77.16% to 99.88%) and 87.1% (95% CI: 70.17% to 96.37%), respectively. Kappa value revealed that the strength of agreement is considered to be very ‘good’. In house established ELISA was implemented in the field and 12.1% (31/256) large ruminants were found positive, while 4.68% (12/256) with postmortem examination. The fasciolosis did not show significant (P>0.05) association with host type, breed and age groups, while significant (P<0.01) association with host sexes. The result confirmed that in-house established ELISA test had good value for serodiagnosis of fasciolosis in cattle and buffaloes for large scale epidemiological studies.
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References
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Anuracpreeda, P., J. Poljaroen, C. Chotwiwatthanakun, Y. Tinikul and P. Sobhon. 2013. Antigenic components, isolation and partial characterization of excretion-secretion fraction of Paramphistomum cervi. Exp. Parasitol., 133(3): 327-333. DOI: 10.1016/j.exppara.2012.12.006
Arias, M., G.V. Hillyer, R. Sánchez-Andrade, J.L. Suárez, J. Pedreira, C. Lomba, P. Díaz, P. Morrondo, P. Díez-Baños and A. Paz-Silva. 2006. A 2.9 kDa Fasciola hepatica-recombinant protein-based ELISA test for the detection of current-ovine fasciolosis trickle infected. Vet. Parasitol., 137(1-2): 67-73. DOI: 10.1016/j.vetpar.2005.12.015
Arora, R., N.K. Singh, P.D. Juyal and S. Ghosh. 2010. Immunoaffinity chromatographic analysis for purification of specific diagnostic antigens of Paramphistomum epiclitum. Journal of Parasitic Diseases, 34(1): 57-61. DOI: 10.1007/s12639-010-0011-6
Awad, W.S., A.K. Ibrahim and F.A. Salib. 2009. Using indirect ELISA to assess different antigens for the serodiagnosis of Fasciola gigantica infection in cattle, sheep and donkeys. Res. Vet. Sci., 86(3): 466-471. DOI: 10.1016/j.rvsc.2008.08.009
Azam, M., M.M. Siddiqui and G. Habib. 2002. Prevalence of parasitic infection in buffalo calves in Khadagzai, district Dir. Pak. Vet. J., 22(2): 87-90. Available on: http://www.pvj.com.pk/pdf-files/22_2/87-90.pdf
Bhutto, B., M.S. Phullan, R. Rind and A.H. Soomro. 2002. Prevalence of gastro-intestinal helminths in buffalo calves. Journal of Biological Sciences, 2(1): 43-45. DOI: 10.3923/jbs.2002.43.45
Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 72(1-2): 248-254. DOI: 10.1016/0003-2697(76)90527-3
De Almeida, M.A., M.B. Ferreira, S. Planchart, A. Terashima, V. Maco, L. Marcos, E. Gotuzzo, E. Sánchez, C. Náquira, J.V. Scorza and R.N. Incani. 2007. Preliminary antigenic characterisation of an adult worm vomit preparation of Fasciola hepatica by infected human sera. Rev. Inst. Med. Trop. Sao Paulo, 49: 31-35. DOI: 10.1590/s0036-46652007000100006
Ferre, I., L.M. Ortega-Mora and F.A. Rojo-Vázquez. 1997. Serum and bile antibody responses (IgG and IgA) during subclinical Fasciola hepatica infection in sheep. Vet. Parasitol., 68(3): 261-267. DOI: 10.1016/s0304-4017(96)01070-9
Gadahi, J.A., M.J. Arshed, Q. Ali, S.B. Javaid and S.I. Shah. 2009. Prevalence of gastrointestinal parasites of sheep and goat in and around Rawalpindi and Islamabad, Pakistan. Vet. World, 2(2): 51-53. Available on: http://citeseerx.ist.psu.edu/viewdoc/download?doi=10.1.1.302.1093&rep=rep1&type=pdf
Gönenç, B., H.O. Sarimehmetoğlu, M. Kara and F. Kircali. 2004. Comparison of crude and excretory/secretory antigens for the diagnosis of Fasciola hepatica in sheep by western blotting. Turk. J. Vet. Anim. Sci., 28: 943-949. Available on: https://journals.tubitak.gov.tr/veterinary/issues/vet-04-28-5/vet-28-5-24-0306-14.pdf
Guobadia, E.E. and B.O. Fagbemi. 1997. The isolation of Fasciola gigantica-specific antigens and their use in the serodiagnosis of fasciolosis in sheep by the detection of circulating antigens. Vet. Parasitol., 68(3): 269-282. DOI: 10.1016/S0304-4017(96)01065-5
Gupta, S.C., S. Ghosh, D.A.V.I.D. Joseph and B.P. Singh. 2003. Diagnosis of experimental Fasciola gigantica infection in cattle by affinity purified antigen. Indian J. Anim. Sci., 7: 963-966.
Hongbao, M. and Sh. Kuan-Jiunn. 2006. Western blotting method. J. Am. Sci., 4: 23-27.
Iqbal, M.U., M.S. Sajid, A. Hussain and M.K. Khan. 2007. Prevalence of helminth infections in dairy animals of Nestle milk collection areas of Punjab (Pakistan). Ital. J. Anim. Sci., 6: 935-938. DOI: 10.4081/ijas.2007.s2.935
Kakar, M.N., M.I. Masood, K.H. Janbaz, M.I. Qadir, I. Masood and J.K. Kakarsulemankhel. 2011. Prevalence of fascioliasis in cows and buffaloes in Quetta, Pakistan. 2: 974-978. Available on: https://pharmacologyonline.silae.it/files/newsletter/2011/vol2/080.qadiir.pdf
Khedri, J., M.H. Radfar, H. Borji and M. Mirzaei. 2015. Prevalence and intensity of Paramphistomum spp. in cattle from south-eastern Iran. Iran. J. Parasitol., 10(2): 268-272.
Kumar, N., S. Ghosh and S.C. Gupta. 2008. Detection of Fasciola gigantica infection in buffaloes by enzyme-linked immunosorbent assay. Parasitol. Res., 104(1): 155-161. DOI: 10.1007/s00436-008-1174-2
Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. International Journal of Science and Nature, 227(5259): 680-685. DOI: 10.1038/227680a0
Mas-Coma, S., M.A. Valero and M.D. Bargues. 2014. Fascioliasis. Adv. Exp. Med. Biol., 766: 77-114.
Mas-Coma, S., M.A. Valero and M.D. Bargues. 2009. Chapter 2. Fasciola, lymnaeids and human fascioliasis, with a global overview on disease transmission, epidemiology, evolutionary genetics, molecular epidemiology and control. Adv. Parasitol., 69: 41-146. DOI: 10.1016/S0065-308X(09)69002-3
Meshgi, B., A. Eslami and F. Hematzadeh. 2008. Determination of somatic and excretory-secretory antigens of Fasciola hepatica and Fasciola gigantica using SDS-PAGE. Iran. J. Vet. Res., 9(1): 77-80. DOI: 10.22099/IJVR.2008.526
Mezo, M., M. Gonza´ lez-Warleta and F.M. Ubeira. 2007. The use of MM3 monoclonal antibodies for the early immunodiagnosis of ovine fascioliasis. J. Parasitol., 93(1): 65-72. DOI: 10.1645/GE-925R.1
Morales, A. and A.M. Espino. 2012. Evaluation and characterization of Fasciola hepatica tegument protein extract for serodiagnosis of human fascioliasis. Clin. Vaccine Immunol., 19(11): 1870-1878. DOI: 10.1128/CVI.00487-12
Muiño, L., M.J. Perteguer, T. Gárate, V. Martínez-Sernández, A. Beltrán, F. Romarís, M. Mezo, M. González-Warleta and F.M. Ubeira. 2011. Molecular and immunological characterization of Fasciola antigens recognized by the MM3 monoclonal antibody. Mol. Biochem. Parasit., 179(2): 80-90. DOI: 10.1016/j.molbiopara.2011.06.003
Phiri, A.M., I.K. Phiri and J. Monrad. 2006. Prevalence of amphistomiasis and its association with Fasciola gigantica infections in Zambian cattle from communal grazing areas. J. Helminthol., 80(1): 65-68. DOI: 10.1079/JOH2005313
Raza, M.A., Z. Iqbal, A. Jabbar and M. Yaseen. 2007. Point prevalence of gastrointestinal helminthiasis in ruminants in southern Punjab, Pakistan. J. Helminthol., 81(3): 323-328. DOI: 10.1017/S0022149X07818554
Rehman, T., M.N. Khan, R.Z. Abbas, W. Babar, A. Sikandar and M.A. Zaman. 2016. Serological and coprological analyses for the diagnosis of Fasciola gigantica infections in bovine hosts from Sargodha, Pakistan. J. Helminthol., 90(4): 494-502. DOI: 10.1017/S0022149X15000711
Salimi-Bejestani, M.R., J.W. McGarry, S. Felstead, P. Ortiz, A. Akca and D.J. Williams. 2005. Development of an antibody-detection ELISA for Fasciola hepatica and its evaluation against a commercially available test. Res. Vet. Sci., 78(2): 177-181. DOI: 10.1016/j.rvsc.2004.08.005
Spithill, T.W. 1999. Pathology, pathophysiology and clinical aspects. p. 185-224. In Dalton, J. (ed.) Fasciolosis. CAB International, London, UK.
Sriveny, D., O.K. Raina, S.C. Yadav, D. Chandra, A.K. Jayraw, M. Singh, R. Velusamy and B.P. Singh. 2006. Cathepsin L cysteine proteinase in the diagnosis of bovine Fasciola gigantica infection. Vet. Parasitol., 135(1): 25-31. DOI: 10.1016/j.vetpar.2005.10.016
Torres, G.B., A.T. Iwashita, C.M. Vargas, L.V. Luján, H.A. Bianchi and R.T. Casanova. 2004. Fasciolosis humana y compromiso gastrointestinal: Estudio de 277 pacientes en el Hospital Nacional Cayetano Heredia. Revista de Gastroenterología del Perú, 24(2): 1970-2002.
Velusamy, R., B.P. Singh, R.L. Sharma and D. Chandra. 2004. Detection of circulating 54kDa antigen in sera of bovine calves experimentally infected with F. gigantica. Vet. Parasitol., 119(2-3): 187-195. DOI: 10.1016/j.vetpar.2003.09.023
Anderson, N., T.T. Luong, N.G. Vo, K.L. Bui, P.M. Smooker and T.W. Spithill. 1999. The sensitivity and specificity of two methods for detecting Fasciola infections in cattle. Vet. Parasitol., 83(1): 15-24. DOI: 10.1016/s0304-4017(99)00026-6
Anuracpreeda, P., J. Poljaroen, C. Chotwiwatthanakun, Y. Tinikul and P. Sobhon. 2013. Antigenic components, isolation and partial characterization of excretion-secretion fraction of Paramphistomum cervi. Exp. Parasitol., 133(3): 327-333. DOI: 10.1016/j.exppara.2012.12.006
Arias, M., G.V. Hillyer, R. Sánchez-Andrade, J.L. Suárez, J. Pedreira, C. Lomba, P. Díaz, P. Morrondo, P. Díez-Baños and A. Paz-Silva. 2006. A 2.9 kDa Fasciola hepatica-recombinant protein-based ELISA test for the detection of current-ovine fasciolosis trickle infected. Vet. Parasitol., 137(1-2): 67-73. DOI: 10.1016/j.vetpar.2005.12.015
Arora, R., N.K. Singh, P.D. Juyal and S. Ghosh. 2010. Immunoaffinity chromatographic analysis for purification of specific diagnostic antigens of Paramphistomum epiclitum. Journal of Parasitic Diseases, 34(1): 57-61. DOI: 10.1007/s12639-010-0011-6
Awad, W.S., A.K. Ibrahim and F.A. Salib. 2009. Using indirect ELISA to assess different antigens for the serodiagnosis of Fasciola gigantica infection in cattle, sheep and donkeys. Res. Vet. Sci., 86(3): 466-471. DOI: 10.1016/j.rvsc.2008.08.009
Azam, M., M.M. Siddiqui and G. Habib. 2002. Prevalence of parasitic infection in buffalo calves in Khadagzai, district Dir. Pak. Vet. J., 22(2): 87-90. Available on: http://www.pvj.com.pk/pdf-files/22_2/87-90.pdf
Bhutto, B., M.S. Phullan, R. Rind and A.H. Soomro. 2002. Prevalence of gastro-intestinal helminths in buffalo calves. Journal of Biological Sciences, 2(1): 43-45. DOI: 10.3923/jbs.2002.43.45
Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 72(1-2): 248-254. DOI: 10.1016/0003-2697(76)90527-3
De Almeida, M.A., M.B. Ferreira, S. Planchart, A. Terashima, V. Maco, L. Marcos, E. Gotuzzo, E. Sánchez, C. Náquira, J.V. Scorza and R.N. Incani. 2007. Preliminary antigenic characterisation of an adult worm vomit preparation of Fasciola hepatica by infected human sera. Rev. Inst. Med. Trop. Sao Paulo, 49: 31-35. DOI: 10.1590/s0036-46652007000100006
Ferre, I., L.M. Ortega-Mora and F.A. Rojo-Vázquez. 1997. Serum and bile antibody responses (IgG and IgA) during subclinical Fasciola hepatica infection in sheep. Vet. Parasitol., 68(3): 261-267. DOI: 10.1016/s0304-4017(96)01070-9
Gadahi, J.A., M.J. Arshed, Q. Ali, S.B. Javaid and S.I. Shah. 2009. Prevalence of gastrointestinal parasites of sheep and goat in and around Rawalpindi and Islamabad, Pakistan. Vet. World, 2(2): 51-53. Available on: http://citeseerx.ist.psu.edu/viewdoc/download?doi=10.1.1.302.1093&rep=rep1&type=pdf
Gönenç, B., H.O. Sarimehmetoğlu, M. Kara and F. Kircali. 2004. Comparison of crude and excretory/secretory antigens for the diagnosis of Fasciola hepatica in sheep by western blotting. Turk. J. Vet. Anim. Sci., 28: 943-949. Available on: https://journals.tubitak.gov.tr/veterinary/issues/vet-04-28-5/vet-28-5-24-0306-14.pdf
Guobadia, E.E. and B.O. Fagbemi. 1997. The isolation of Fasciola gigantica-specific antigens and their use in the serodiagnosis of fasciolosis in sheep by the detection of circulating antigens. Vet. Parasitol., 68(3): 269-282. DOI: 10.1016/S0304-4017(96)01065-5
Gupta, S.C., S. Ghosh, D.A.V.I.D. Joseph and B.P. Singh. 2003. Diagnosis of experimental Fasciola gigantica infection in cattle by affinity purified antigen. Indian J. Anim. Sci., 7: 963-966.
Hongbao, M. and Sh. Kuan-Jiunn. 2006. Western blotting method. J. Am. Sci., 4: 23-27.
Iqbal, M.U., M.S. Sajid, A. Hussain and M.K. Khan. 2007. Prevalence of helminth infections in dairy animals of Nestle milk collection areas of Punjab (Pakistan). Ital. J. Anim. Sci., 6: 935-938. DOI: 10.4081/ijas.2007.s2.935
Kakar, M.N., M.I. Masood, K.H. Janbaz, M.I. Qadir, I. Masood and J.K. Kakarsulemankhel. 2011. Prevalence of fascioliasis in cows and buffaloes in Quetta, Pakistan. 2: 974-978. Available on: https://pharmacologyonline.silae.it/files/newsletter/2011/vol2/080.qadiir.pdf
Khedri, J., M.H. Radfar, H. Borji and M. Mirzaei. 2015. Prevalence and intensity of Paramphistomum spp. in cattle from south-eastern Iran. Iran. J. Parasitol., 10(2): 268-272.
Kumar, N., S. Ghosh and S.C. Gupta. 2008. Detection of Fasciola gigantica infection in buffaloes by enzyme-linked immunosorbent assay. Parasitol. Res., 104(1): 155-161. DOI: 10.1007/s00436-008-1174-2
Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. International Journal of Science and Nature, 227(5259): 680-685. DOI: 10.1038/227680a0
Mas-Coma, S., M.A. Valero and M.D. Bargues. 2014. Fascioliasis. Adv. Exp. Med. Biol., 766: 77-114.
Mas-Coma, S., M.A. Valero and M.D. Bargues. 2009. Chapter 2. Fasciola, lymnaeids and human fascioliasis, with a global overview on disease transmission, epidemiology, evolutionary genetics, molecular epidemiology and control. Adv. Parasitol., 69: 41-146. DOI: 10.1016/S0065-308X(09)69002-3
Meshgi, B., A. Eslami and F. Hematzadeh. 2008. Determination of somatic and excretory-secretory antigens of Fasciola hepatica and Fasciola gigantica using SDS-PAGE. Iran. J. Vet. Res., 9(1): 77-80. DOI: 10.22099/IJVR.2008.526
Mezo, M., M. Gonza´ lez-Warleta and F.M. Ubeira. 2007. The use of MM3 monoclonal antibodies for the early immunodiagnosis of ovine fascioliasis. J. Parasitol., 93(1): 65-72. DOI: 10.1645/GE-925R.1
Morales, A. and A.M. Espino. 2012. Evaluation and characterization of Fasciola hepatica tegument protein extract for serodiagnosis of human fascioliasis. Clin. Vaccine Immunol., 19(11): 1870-1878. DOI: 10.1128/CVI.00487-12
Muiño, L., M.J. Perteguer, T. Gárate, V. Martínez-Sernández, A. Beltrán, F. Romarís, M. Mezo, M. González-Warleta and F.M. Ubeira. 2011. Molecular and immunological characterization of Fasciola antigens recognized by the MM3 monoclonal antibody. Mol. Biochem. Parasit., 179(2): 80-90. DOI: 10.1016/j.molbiopara.2011.06.003
Phiri, A.M., I.K. Phiri and J. Monrad. 2006. Prevalence of amphistomiasis and its association with Fasciola gigantica infections in Zambian cattle from communal grazing areas. J. Helminthol., 80(1): 65-68. DOI: 10.1079/JOH2005313
Raza, M.A., Z. Iqbal, A. Jabbar and M. Yaseen. 2007. Point prevalence of gastrointestinal helminthiasis in ruminants in southern Punjab, Pakistan. J. Helminthol., 81(3): 323-328. DOI: 10.1017/S0022149X07818554
Rehman, T., M.N. Khan, R.Z. Abbas, W. Babar, A. Sikandar and M.A. Zaman. 2016. Serological and coprological analyses for the diagnosis of Fasciola gigantica infections in bovine hosts from Sargodha, Pakistan. J. Helminthol., 90(4): 494-502. DOI: 10.1017/S0022149X15000711
Salimi-Bejestani, M.R., J.W. McGarry, S. Felstead, P. Ortiz, A. Akca and D.J. Williams. 2005. Development of an antibody-detection ELISA for Fasciola hepatica and its evaluation against a commercially available test. Res. Vet. Sci., 78(2): 177-181. DOI: 10.1016/j.rvsc.2004.08.005
Spithill, T.W. 1999. Pathology, pathophysiology and clinical aspects. p. 185-224. In Dalton, J. (ed.) Fasciolosis. CAB International, London, UK.
Sriveny, D., O.K. Raina, S.C. Yadav, D. Chandra, A.K. Jayraw, M. Singh, R. Velusamy and B.P. Singh. 2006. Cathepsin L cysteine proteinase in the diagnosis of bovine Fasciola gigantica infection. Vet. Parasitol., 135(1): 25-31. DOI: 10.1016/j.vetpar.2005.10.016
Torres, G.B., A.T. Iwashita, C.M. Vargas, L.V. Luján, H.A. Bianchi and R.T. Casanova. 2004. Fasciolosis humana y compromiso gastrointestinal: Estudio de 277 pacientes en el Hospital Nacional Cayetano Heredia. Revista de Gastroenterología del Perú, 24(2): 1970-2002.
Velusamy, R., B.P. Singh, R.L. Sharma and D. Chandra. 2004. Detection of circulating 54kDa antigen in sera of bovine calves experimentally infected with F. gigantica. Vet. Parasitol., 119(2-3): 187-195. DOI: 10.1016/j.vetpar.2003.09.023
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Published
2022-03-25
How to Cite
Afshan, K., Sajid, M., Komal, M., Hassan, H. S. Z. ul, Narjis, G., & Firasat, S. (2022). Diagnostic potential of 36-55 kDa somatic antigens of Fasciola gigantica for bovine fasciolosis. Buffalo Bulletin, 41(1), 49–61. https://doi.org/10.56825/bufbu.2022.4113170
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