Skip to main content
Top
Published in: Journal of Parasitic Diseases 4/2017

01-12-2017 | Original Article

Efficacy of glutathione-S-transferase purified antigen of the gastro-intestinal nematode Haemonchus contortus in diagnosis of sheep haemonchosis

Authors: Omnia M. Kandil, Irene S. Gamil, Seham H. M. Hendawy, Fatma Medhat, Ola H. El-Habit

Published in: Journal of Parasitic Diseases | Issue 4/2017

Login to get access

Abstract

Haemonchus contortus is one of the most pathogenic and economically important parasites of sheep. Different H. contortus antigens; crude somatic antigen (CSA), excretory/secretory antigen (ESA), crude larval antigen (CLA), glutathione-S-transferase antigen (GST) and recombinant protein (rhcp 26/23) were prepared and characterized using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot. The antigens were immunologically evaluated through indirect enzyme linked immunosorbent assay (ELISA) for diagnosis of haemonchosis in experimentally and naturally infected sheep. Analysis of the resultant bands of SDS-PAGE demonstrated that 13, 6, 11, 2 and 1 protein bands from CSA, ESA, CLA, GST and rhcp 26/23, respectively and analysis of the resultant bands of western blot showed that 13, 6, 4 and 1 reactive bands detected from CSA, ESA, CLA and GST, respectively. The results of ELISA of different antigens revealed that sero-prevelance of CSA, ESA, CLA, GST and rhcp 26/23 were 78.51, 82.34, 85.319, 45.319 and 90.8% respectively, sensitivity were 100, 90, 100, 96.66 and 90%, respectively and specificity were 0, 70, 10, 70 and 6.66%, respectively with diagnostic potency were 50, 80, 55, 83.33 and 48.33%, respectively. Statistical analysis using Chi square test found that GST is the best one that can be used. The cross reactivity of GST antigen, crude Fasciola antigen and crude Moniezia antigen tested versus their homologous hyper immune sera at different dilutions using ELISA. The current study reported that GST antigen could be considered as a promising antigen for diagnosis of haemonchosis.
Literature
go back to reference Anderson DV, Dixon SC, Graham RB, Smith WD, Tucker EM (1989) Ovine monoclonal antibody to Ostertagia circumcincta. Biochem Soc Trans 17:736CrossRef Anderson DV, Dixon SC, Graham RB, Smith WD, Tucker EM (1989) Ovine monoclonal antibody to Ostertagia circumcincta. Biochem Soc Trans 17:736CrossRef
go back to reference Angulo-Cubillán FJ, García-Coiradas L, Cuquerella M, Fuente CDL, Alunda YJM (2007) Haemonchus contortus sheep relationship: a review. Rev Cient 11:577–587 Angulo-Cubillán FJ, García-Coiradas L, Cuquerella M, Fuente CDL, Alunda YJM (2007) Haemonchus contortus sheep relationship: a review. Rev Cient 11:577–587
go back to reference Cuquerella M, Gomez-Munoz MT, Carrera L, de la Fuente C (1994) Cross antigenicity among ovine trichostrongyloidea, preliminary report. Vet Parasitol 53:243–251CrossRefPubMed Cuquerella M, Gomez-Munoz MT, Carrera L, de la Fuente C (1994) Cross antigenicity among ovine trichostrongyloidea, preliminary report. Vet Parasitol 53:243–251CrossRefPubMed
go back to reference Fagbemi BO, Obarisiagbon IO, Mbuh JV (1995) Detection of circulating antigen in sera of Fasciola gigantica infected cattle with antibodies reactive with a Fasciola specific 88 kDa antigen. Vet Parasitol 58:235–246CrossRefPubMed Fagbemi BO, Obarisiagbon IO, Mbuh JV (1995) Detection of circulating antigen in sera of Fasciola gigantica infected cattle with antibodies reactive with a Fasciola specific 88 kDa antigen. Vet Parasitol 58:235–246CrossRefPubMed
go back to reference Fawzi EM, González-Sánchez ME, Corral MJ, Cuquerella M, Alunda JM (2015) Vaccination of lambs against Haemonchus contortus infection with a somatic protein (Hc23) from adult helminthes. Int J Parasitol 44:429–436CrossRef Fawzi EM, González-Sánchez ME, Corral MJ, Cuquerella M, Alunda JM (2015) Vaccination of lambs against Haemonchus contortus infection with a somatic protein (Hc23) from adult helminthes. Int J Parasitol 44:429–436CrossRef
go back to reference García-Coiradas L, Angulo-Cubillán F, Valladares B, Martínez E, de la Fuente C, Alunda JM, Cuquerella M (2010) Immunization against lamb haemonchosis with a recombinant somatic antigen of Haemonchus contortus (rHcp26/23). Vet Med Int 2010:852146. doi:10.4061/2010/852146 CrossRefPubMedPubMedCentral García-Coiradas L, Angulo-Cubillán F, Valladares B, Martínez E, de la Fuente C, Alunda JM, Cuquerella M (2010) Immunization against lamb haemonchosis with a recombinant somatic antigen of Haemonchus contortus (rHcp26/23). Vet Med Int 2010:852146. doi:10.​4061/​2010/​852146 CrossRefPubMedPubMedCentral
go back to reference Gill HS (1991) Genetic control of acquired resistance to haemonchosis in Merino lambs. Parasite Immunol 13:617–628CrossRefPubMed Gill HS (1991) Genetic control of acquired resistance to haemonchosis in Merino lambs. Parasite Immunol 13:617–628CrossRefPubMed
go back to reference Habig WH, Jakoby WB (1981) Assay for differentiation of glutathione s-transferases. Methods Enzymol 77:398–405CrossRefPubMed Habig WH, Jakoby WB (1981) Assay for differentiation of glutathione s-transferases. Methods Enzymol 77:398–405CrossRefPubMed
go back to reference Hudson L, Hay F (1989) Pract Immunol, 3rd edn. Blackwell, Oxford Hudson L, Hay F (1989) Pract Immunol, 3rd edn. Blackwell, Oxford
go back to reference Johnson DA, Behnke JM, Coles GC (2004) Copro-antigen capture EL1SA for the detection of Teladorsagia (Ostertagia) circumcincta in sheep: improvement of specificity by heat treatment. Parasitol 129:115–126CrossRef Johnson DA, Behnke JM, Coles GC (2004) Copro-antigen capture EL1SA for the detection of Teladorsagia (Ostertagia) circumcincta in sheep: improvement of specificity by heat treatment. Parasitol 129:115–126CrossRef
go back to reference Kandil OM, Eid NA, Elakabawy LM, Abdelrahman KA, Helal MA (2015) Immunodiagnostic potency of different Haemonchus contortus antigens for diagnosis of experimentally and naturally haemonchosis in Egyptian sheep. APG 6:238–247 Kandil OM, Eid NA, Elakabawy LM, Abdelrahman KA, Helal MA (2015) Immunodiagnostic potency of different Haemonchus contortus antigens for diagnosis of experimentally and naturally haemonchosis in Egyptian sheep. APG 6:238–247
go back to reference Kandil OM, Hendawy SHM, El Namaky AH, Gabrashanska MP, Nanev VN (2016) Evaluation of different Haemonchus contortus antigens for diagnosis of sheep haemonchosis by ELISA and their cross reactivity with other helminthes. J Parasit Dis. doi:10.1007/s12639-016-0865-3 Kandil OM, Hendawy SHM, El Namaky AH, Gabrashanska MP, Nanev VN (2016) Evaluation of different Haemonchus contortus antigens for diagnosis of sheep haemonchosis by ELISA and their cross reactivity with other helminthes. J Parasit Dis. doi:10.​1007/​s12639-016-0865-3
go back to reference Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefPubMed Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefPubMed
go back to reference Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275PubMed Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275PubMed
go back to reference MAFF (Ministry of Agriculture, Fisheries and Food) (1986) Manual of veterinary parasitological laboratory techniques, 3rd edn. HSMO Publications, London, UK, 160 pp MAFF (Ministry of Agriculture, Fisheries and Food) (1986) Manual of veterinary parasitological laboratory techniques, 3rd edn. HSMO Publications, London, UK, 160 pp
go back to reference Maizels RM, Blaxter ML, Robertson BD, Selkirk ME (1991) Parasite antigens, parasite genes: a laboratory manual for molecular parasitology, 1st edn. Cambridge University Press, London Maizels RM, Blaxter ML, Robertson BD, Selkirk ME (1991) Parasite antigens, parasite genes: a laboratory manual for molecular parasitology, 1st edn. Cambridge University Press, London
go back to reference McKerrow JH, Brindley P, Brown M, Gam AA, Staunton C, Neva FA (1990) Strongyloidesstercoralis: identification of a protease that facilitates penetration of skin by the infective larvae. Exp Parasitol 70:134–143CrossRefPubMed McKerrow JH, Brindley P, Brown M, Gam AA, Staunton C, Neva FA (1990) Strongyloidesstercoralis: identification of a protease that facilitates penetration of skin by the infective larvae. Exp Parasitol 70:134–143CrossRefPubMed
go back to reference Molina JM, Ruiz A, Rodríguez-Ponce E, Gutiérrez AC, González J, Hernández S (1999) Cross reactive antigens of Haemonchus contortus adult worms in Teladorsagia circumcincta infected goats. Vet Res 30:393–399PubMed Molina JM, Ruiz A, Rodríguez-Ponce E, Gutiérrez AC, González J, Hernández S (1999) Cross reactive antigens of Haemonchus contortus adult worms in Teladorsagia circumcincta infected goats. Vet Res 30:393–399PubMed
go back to reference Pour LM, Farahnak A, Rad MM, Golmohamadi T, Eshraghian M (2014) Activity assay of glutathione s-transferase (GSTs) enzyme as a diagnostic biomarker for liver hydatid cyst in vitro. Iran J Public Health 43:994–999 Pour LM, Farahnak A, Rad MM, Golmohamadi T, Eshraghian M (2014) Activity assay of glutathione s-transferase (GSTs) enzyme as a diagnostic biomarker for liver hydatid cyst in vitro. Iran J Public Health 43:994–999
go back to reference Prasad A, Nasir A, Singh N (2008) Detection of anti-Haemonchus contortus antibodies in sheep by dot-ELISA with immune-affinity purified fraction of ES antigen during prepatency. Indian J Exp Biol 46:94–99PubMed Prasad A, Nasir A, Singh N (2008) Detection of anti-Haemonchus contortus antibodies in sheep by dot-ELISA with immune-affinity purified fraction of ES antigen during prepatency. Indian J Exp Biol 46:94–99PubMed
go back to reference Qamar MF, Maqbool A (2012) Biochemical studies and serodiagnosis of haemonchosis in sheep and goats. J Anim Plant Sci 22:32–38 Qamar MF, Maqbool A (2012) Biochemical studies and serodiagnosis of haemonchosis in sheep and goats. J Anim Plant Sci 22:32–38
go back to reference Richer JK, Sakanari JA, Frank GR, Grieve RB (1992) Dirofilaria immitis: proteases produced by third- and fourth-stage larvae. Exp Parasitol 75:213–222CrossRefPubMed Richer JK, Sakanari JA, Frank GR, Grieve RB (1992) Dirofilaria immitis: proteases produced by third- and fourth-stage larvae. Exp Parasitol 75:213–222CrossRefPubMed
go back to reference Salama DB (2017) Immunization of sheep against Haemonchus contortus worms. M.V.Sc. Thesis, Faculty of Veterinary Medicine, Beni-Suef University, Egypt Salama DB (2017) Immunization of sheep against Haemonchus contortus worms. M.V.Sc. Thesis, Faculty of Veterinary Medicine, Beni-Suef University, Egypt
go back to reference Schallig HDFH, Hornok S, Cornelissen JBWJ (1995a) Comparison of two enzyme immunoassays for the detection of Haemonchus contortus infections in sheep. Vet Parasitol 57:329–338CrossRefPubMed Schallig HDFH, Hornok S, Cornelissen JBWJ (1995a) Comparison of two enzyme immunoassays for the detection of Haemonchus contortus infections in sheep. Vet Parasitol 57:329–338CrossRefPubMed
go back to reference Schallig HD, van Leeuwen MA, Hendrikx WM (1995b) Isotype-specific serum antibody responses of sheep to Haemonchus contortus antigens. Vet Parasitol 56:149–162CrossRefPubMed Schallig HD, van Leeuwen MA, Hendrikx WM (1995b) Isotype-specific serum antibody responses of sheep to Haemonchus contortus antigens. Vet Parasitol 56:149–162CrossRefPubMed
go back to reference Tariq KA, Chishti MZ, Fayaz A, Shawl AS (2008) Epidemiology of gastrointestinal nematodes of sheep managed under traditional husbandry system in Kashmir valley. Vet Parasitol 158:138–143CrossRefPubMed Tariq KA, Chishti MZ, Fayaz A, Shawl AS (2008) Epidemiology of gastrointestinal nematodes of sheep managed under traditional husbandry system in Kashmir valley. Vet Parasitol 158:138–143CrossRefPubMed
go back to reference Thrusfield M (1997) Veterinary epidemiology, 2nd edn. Blackwell Science Ltd., Oxford, pp 134–135 Thrusfield M (1997) Veterinary epidemiology, 2nd edn. Blackwell Science Ltd., Oxford, pp 134–135
go back to reference Towbin H, Staeheline T, Gordon J (1979) Electrophoretic transfer of protein from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci 76:4350–4354CrossRefPubMedPubMedCentral Towbin H, Staeheline T, Gordon J (1979) Electrophoretic transfer of protein from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci 76:4350–4354CrossRefPubMedPubMedCentral
go back to reference Wijffels GL, Sexton JL, Salvatore L, Pettitt JM, Humphris DC, Panaccio M, Spithill TW (1992) Primary sequence heterogeneity and tissue expression of glutathione S-transferases of Fasciola hepatica. Exp Parasitol 74:87–99CrossRefPubMed Wijffels GL, Sexton JL, Salvatore L, Pettitt JM, Humphris DC, Panaccio M, Spithill TW (1992) Primary sequence heterogeneity and tissue expression of glutathione S-transferases of Fasciola hepatica. Exp Parasitol 74:87–99CrossRefPubMed
Metadata
Title
Efficacy of glutathione-S-transferase purified antigen of the gastro-intestinal nematode Haemonchus contortus in diagnosis of sheep haemonchosis
Authors
Omnia M. Kandil
Irene S. Gamil
Seham H. M. Hendawy
Fatma Medhat
Ola H. El-Habit
Publication date
01-12-2017
Publisher
Springer India
Published in
Journal of Parasitic Diseases / Issue 4/2017
Print ISSN: 0971-7196
Electronic ISSN: 0975-0703
DOI
https://doi.org/10.1007/s12639-017-0920-8

Other articles of this Issue 4/2017

Journal of Parasitic Diseases 4/2017 Go to the issue
Live Webinar | 27-06-2024 | 18:00 (CEST)

Keynote webinar | Spotlight on medication adherence

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

WHO estimates that half of all patients worldwide are non-adherent to their prescribed medication. The consequences of poor adherence can be catastrophic, on both the individual and population level.

Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.

Prof. Kevin Dolgin
Prof. Florian Limbourg
Prof. Anoop Chauhan
Developed by: Springer Medicine
Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Watch this official video from ACC.24. Dr. Biykem Bozkurt discusses last year's major advances in heart failure and cardiomyopathies.