2D electrophoretic profile of Fasciola hepatica intestine proteins

• Marco Cabrera-González Instituto Nacional de Innovación Agraria (INIA), Dirección de Desarrollo Tecnológico Agrario, Estación Experimental Baños del Inca. Baños del Inca, Cajamarca, Perú
• Diana Marlo-Javier Instituto Nacional de Innovación Agraria (INIA), Dirección de Desarrollo Tecnológico Agrario, Estación Experimental Baños del Inca. Baños del Inca, Cajamarca, Perú
• Carlos Quilcate-Pairazamán Instituto Nacional de Innovación Agraria (INIA), Dirección de Desarrollo Tecnológico Agrario. La Molina, Lima, Perú
• Héctor V. Vásquez Instituto Nacional de Innovación Agraria (INIA), Dirección de Desarrollo Tecnológico Agrario. La Molina, Lima, Perú
• Medali Cueva-Rodríguez Instituto Nacional de Innovación Agraria (INIA), Dirección de Desarrollo Tecnológico Agrario, Estación Experimental Baños del Inca. Baños del Inca, Cajamarca, Perú

DOI: https://doi.org/10.52973/rcfcv-e32103

Keywords: Bovine, fasciolosis, 2D electrophoresis, parasites

Abstract

Being fasciolosis an important parasitic infection in ruminants in many Countries and given the high prevalence in humans and animals in Cajamarca, Peru. It was proposed to carry out the study on the profile of the intestinal proteins of Fasciola hepatica with the objective of knowing the number of proteins and the pH range of secretion/excretion of the intestine of the parasite that it expresses by means of the 2D-bidimensional electrophoresis method. Adult samples of F. hepatica were collected from bovine livers in the Cajamarca Municipal Slaughterhouse. They were transferred to the Animal Health Biotechnology Laboratory of the Baños del Inca Agrarian Experimental Station, INIA – Cajamarca, for processing. The electrophoretic run allowed to separate 82 proteins with different molecular weights, focused at different isoelectric points in a pH range of 6.0 – 9.4. It is concluded that by analyzing the 2D gel of F. hepatica gut proteins, 84 protein spots with different molecular weights were identified, focused on different isoelectric points in a range of 6.0 – 9.4.

References

ADDY, F.; GYAN, K.; ARHIN, E.; WASSERMANN, M. Prevalence of bovine fasciolosis from the Bolgatanga abattoir. Scientific. African. 8:e00469. 2020. https://doi.org/htc6.

ANIMAL PLANT HEALTH AGENCY. APHA Parasitology Group. Annual Review of Literature and Horizon Scanning Report 2017. 2018. https://bit.ly/3wcUsY1.

BECERRO-RECIO, D.; GONZÁLEZ-MIGUEL, J.; UCERO, A.; SOTILLO, J.; MARTÍNEZ-MORENO, Á.; PÉREZ-ARÉVALO, J.; CWIKLINSKI, K.; DALTON, J.P.; SILES-LUCAS, M. Recognition Pattern of the Fasciola hepatica Excretome/Secretome during the Course of an Experimental Infection in Sheep by 2D Immunoproteomics. Pathog. 10: 1-11. 2021. https://doi.org/gkhgjq.

BEESLEY, N.J.; ATTREE, E.; VÁZQUEZ-PRIETO, S.; VILAS, R.; PANIAGUA, E.; UBEIRA, F.M.; JENSEN, O.; PRUZZO, C.; ÁLVAREZ, J.D.; MALANDRINI, J.B.; SOLANA, H.; HODGKINSON, J.E. Evidence of population structuring following population genetic analyses of Fasciola hepatica from Argentina. Internat. J. Parasitol. 51(6): 471-480. 2020. https://doi.org/htj6.

BENNETT, A.P.; ROBINSON, M.W. Trematode Proteomics: Recent Advances and Future Directions. Pathog. 10: 348. 2021. https://doi.org/htj7.

BROCKWELL, Y.M.; ELLIOTT, T.P.; ANDERSON, G.R.; STANTON, R.; SPITHILL, T.W.; SANGSTER, N.C. Confirmation of Fasciola hepatica resistant to triclabendazole in naturally infected Australian beef and dairy cattle. Internat. J. Parasitol. 4(1): 48-54. 2014. https://doi.org/htj8.

CALVANI, N.E.D.; ICHIKAWA-SEKI, M.; BUSH, R. D.; KHOUNSY, S.; ŠLAPETA, J. Which species is in the faeces at a time of global livestock movements: single nucleotide polymorphism genotyping assays for the differentiation of Fasciola spp. Internat. J. Parasitol. 50(2): 91-101. 2020. https://doi.org/htj9

CORRAL-RUIZ, G.M; SÁNCHEZ-TORRES, L.E. Fasciola hepatica-derived molecules as potential immunomodulators. Acta Trop. 210:e105548. 2020. https://doi.org/htkb.

CUELLO, M.; CABRERA, O.; RODRÍGUEZ, Y.; THURHEIM, G.; NORHEIM, G.; CABRERA, R.A.; ÁLVAREZ, M.; BLAIN, K.; NAESS, L.; ROSENQVIST, E.; GARCÍA, L. SDS-Page y analisis densitometrico para determinar la concentracion de lipopolisacaridos de Nesseria meningitidis serogrupos A, W135 y X. VacciMonitor. 22(3): 36-42. 2013. https://bit.ly/38Ign1F.

CWIKLINSKI, K.; ALLEN, K.; LACOURSE, J.; WILLIAMS, D.J.; PATERSON, S.; HODGKINSON, J.E. Characterisation of a novel panel of polymorphic microsatellite loci for the liver fluke, Fasciola hepatica ,using a next generation sequencing approach q. Infection. Infect. Genet. Evol. 32: 298-304. 2015. https://doi.org/f7dtfb.

CWIKLINSKI, K.; DALTON, J.P. Advances in Fasciola hepatica research using 'omics' technologies. Internat. J. Parasitol. 48(5): 321-331.2018. https://doi.org/gdg2wg.

DALTON, J.P.; HEFFERNAN, M.Thiol proteases released in vitro by Fasciola hepatica. Molec. Biochem. Parasitol. 35(2): 161-166. 1989. https://doi.org/c39bj7.

DALTON, J.P.; ROBINSON, M.W.; MULCAHY, G.; O’NEILL, S.M.; DONNELLY, S. Immunomodulatory molecules of Fasciola hepatica: Candidates for both vaccine and immunotherapeutic development. Vet. Parasitol. 195: 272-285. 2013. https://doi.org/hvxt.

DORRI, Y. Two-Dimensional Gel Electrophoresis: Vertical Isoelectric Focusing. Meth. Mol Biol. 1855: 291-302. 2019. https://doi.org/hvxw.

ESCALANTE, H.; DAVELOIS, K.; ORTIZ, P.; RODRÍGUEZ, H.; DIAZ, E.; JARA, C. Estandarización de la técnica de Western Blot para el diagnóstico de la fasciolosis humana utilizando antigenos de excreción-secreción de Fasciola hepatica. Rev. Peru Med. Exp. Salud Publ. 28: 454-461. 2011. https://doi.org/fd42s3.

GOURBAL, B.E.; GUILLOU, F.; MITTA, G.; SIBILLE, P.;THÈRON, A.; POINTIER, J.P.; COUSTAU, C. Excretory-secretory products of larval Fasciola hepatica investigated using a two-dimensional proteomic approach. Molec. Biochem. Parasitol. 161(1): 63-66. 2008. https://doi.org/dkjh6g.

GRAPHPAD. 2021. En línea. https://bit.ly/3wzrFOJ. 12-11-2021.

GELANALYZER 2010. Freeware 1D gel electrophoresis image analysis software. 2010. En línea. https://bit.ly/3sK3laL. 23-02-2022.

HAMILTON, C.M.; DOWLING, D.J.; LOSCHER, C.E.; MORPHEW, R.M.; BROPHY, P.M.; O’NEILL, S.M. The Fasciola hepatica Tegumental Antigen Suppresses Dendritic Cell Maturation and Function. Infect. Immunity. 77(6): 2488-2498. 2009. https://doi.org/fs4fxf.

HANNAN-KHAN, M.A.; ULLAH, R.; REHMAN, A.; REHMAN, L.; SHAREEF, A.; ABIDI, S. M. Immunolocalization and immunodetection of the excretory/secretory (ES) antigens of Fasciola gigantica. PLoS ONE. 12: 1-15. 2017. https://doi.org/gbzrnx.

ISAH, U.M. Studies on the prevalence of fascioliasis among ruminant animals in Northern Bauchi Sate, North-Eastern Nigeria. Parasite. Epidemiol. Control. 5: e00090. 2019. https://doi.org/hvxx.

ISSAQ, H.; VEENSTRA, T. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE): advances and perspectives. Biotech. 44: 697-700. 2008. https://doi.org/dwrfqc.

JEFFERIES, J.R.; CAMPBELL, A.M.; ROSSUM, A.J; BARRETT, J.; BROPHY, P.M. Proteomic analysis of Fasciola hepatica excretory-secretory products. Proteomics. 1:1128–1132. 2001. https://doi.org/b8bmrj.

KHAN, Y.A.; KHAN, M.H.; ABIDI, S.M. 2D-PAGE analysis of the soluble proteins of the tropical liver fluke, Fasciola gigantica and Biliary amphistome, Gigantocotyle explanatum, concurrently infecting Bubalus bubalis. J. Parasit. Dis. 40(3): 910-913. 2016. https://doi.org/hvxz.

KOWALCZYK, S.J.; CZOPOWICZ, M.; WEBER, C.N.; MÜLLER, E.; KABA, J. Accuracy of a diagnostic model based on serum biochemical parameters in detecting cows at an increased risk of chronic fascioliasis. Vet. Parasitol. 254: 15-20. 2018. https://doi.org/gdjnfq.

LEE, C.G.; ZIMMERMAN, G.L.; WEE, S.H. Fasciola hepatica: comparison of flukes from Korea and the United States by isoelectric focusing banding patterns of whole-body protein. Vet. Parasitol. 42: 311-316. 1992. https://doi.org/bt37rn.

MAS-COMA, S.; BARGUES, M.D.; VALERO, M.A. Human fascioliasis infection sources, their diversity, incidence factors, analytical methods and prevention measures. Parasitol. 145 (13): 1665-1699. 2018. https://doi.org/gdtz6p.

MIRANDA-MIRANDA, E.; COSSIO-BAYUGAR, R.; AGUILAR-DÍAZ, H.; NARVÁEZ-PADILLA, V.; SACHMAN-RUÍZ, B.; REYNAUD, E. Transcriptome assembly dataset of anthelmintic response in Fasciola hepatica. Data Brief. 35: 106808. 2021. https://doi.org/hvzb.

MOLINA-HERNÁNDEZ, V.; MULCAHY, G.; PÉREZ, J.; MARTÍNEZ-MORENO, Á.; DONNELLY, S.; O’NEILL, S.M.; DALTON, J.P.; CWIKLINSKI, K. Fasciola hepatica vaccine: We may not be there yet but we’re on the right road. Vet. Parasitol. 208 (1-2): 101-111. 2015. https://doi.org/f65jm6.

MORENO, A.; OUTERELO, R.; RUIZ, E.; AGUIRRE, J.; ALMODÓVAR, A.; ALONSO, J.; BENITO, J.; ARILLO, A.; CANO, J. Prácticas de Zoología. Estudio y diversidad de los Platelmintos, Nematodos, Nematomorfos y Acantocéfalos. RE. Serie Zoología. 4(2): 37-60. 2011. https://bit.ly/3yNxjyf.

MORPHEW, R.M.; WILKINSON, T.J.; MACKINTOSH, N.; JAHNDEL, V.; PATERSON, S.; MCVEIGH, P.; ABIDI, S.M.; SAIFULLAH, K.; RAMAN, M.; RAVIKUMAR, G.; LACOURSE, J., MAULE, A.; BROPHY, P.M. Exploring and Expanding the Fatty-Acid-Binding Protein Superfamily in Fasciola Species. Proteome Res. 15: 3308-3321. 2016. https://doi.org/hvzn.

MORPHEW, R.M.; WRIGHT, H.A.; LACOURSE, E.J.; WOODS, D.J.; BROPHY, P.M. Comparative proteomics of excretory-secretory proteins released by the liver fluke Fasciola hepatica in sheep host bile and during in vitro culture ex host. Molec. Cell. Proteomics. 6(6): 963-972. 2007. https://doi.org/d65cs9.

MUÑOZ, M.E.; PLACENCIA, M.; DEL POZO, J.A.; SEVILLA, C.; HUIZA, A. Diagnóstico serológico de la infección por Fasciola hepatica: una revisión sistemática. Rev. Gastroenterol. Perú. 40(2): 155-161. 2020. https://bit.ly/3G8rGwv.

PEREA, M.; DÍAZ, A.; MEDELLÍN, M.; BULLA, D. Fasciolosis: una enfermedad emergente. Pensamiento y Acción. 24: 55-66. 2018. https://bit.ly/38Fixio.

PÉREZ-CABALLERO, R.; SILES-LUCAS, M.; GONZÁLEZ-MIGUEL, J.; MARTÍNEZ-MORENO, F.J.; ESCAMILLA, A.; PÉREZ, J.; MARTÍNEZ-MORENO, A.; BUFFONI, L. Pathological, immunological and parasitological study of sheep vaccinated with the recombinant protein 14-3-3z and experimentally infected with Fasciola hepatica. Vet. Immunol. Immunopathol. 202: 115-121. 2018. https://doi.org/hvzs.

PINILLA, J.C.; FLOREZ, A.A.; ORLANDONI, G.; TOBÓN, J.C.; ORTÍZ, D. Current status of prevalence and risk factors associated with liver fluke Fasciola hepatica in cattle raised in different altitudinal regions of Colombia. Vet. Parasitol: Regional Studies and Reports. 22: 1-6. 2020. https://doi.org/hvzp.

RASOULI, A.; FARAHNAK, A.; ZALI, H.; REZAEIAN, M.; GOLESTANI, A.; MOLAEI, M.B. Protein detection of excretory-secretory products and somatic extracts from Fasciola hepatica and F. gigantica using two-dimensional electrophoresis. Iranian J. Parasitol. 14(3): 379-386. 2019. https://doi.org/hvzr.

ROBINSON, M.W.; MENON, R.; DONNELLY, S.M.; DALTON, J.P.; RANGANATHAN, S. An integrated transcriptomics and proteomics analysis of the secretome of the helminth pathogen Fasciola hepatica: Proteins associated with invasion and infection of the mammalian host. Molec. Cell. Proteomics 8(8): 1891-1907. 2009. https://doi.org/db7kvz.

RYAN, S.; SHIELS, J.; TAGGART, C.C.; DALTON, J.P.; WELDON, S. Fasciola hepatica-Derived Molecules as Regulators of the Host Immune Response. Frontiers Immunol. 11: 2182. 2020. https://doi.org/hvzv.

SMITH, A.M.; DOWD, A.J.; HEFFERNAN, M.; ROBERTSON, C.D.; DALTON, J.P. Fasciola hepatica: A secreted cathepsin L-like proteinase cleaves host immunoglobulin. Internat. J. Parasitol. 23(8): 977-983. 1993. https://doi.org/dzj8ff.

THOMAS, A.P. Memoirs: The Life History of the Liver-Fluke (Fasciola hepatica). J. Cell. Sci. 23(89): 99-133. 1883. https://doi.org/hvzw.

XU, J.; WU, L.; SUN, Y.; WEI, Y.; ZHENG, L.; ZHANG, J.; PANG, Z.; YANG, Y.; LU, Y. Proteomics and bioinformatics analysis of Fasciola hepatica somatic proteome in different growth phases. Parasitol. Res. 119(9): 2837-2850.2020. https://doi.org/hvzx.

YOUNG, N.D.; JEX, A.R.; CANTACESSI, C.; HALL, R.S.; CAMPBELL, B.E.; SPITHILL, T.W.; TANGKAWATTANA, S.; TANGKAWATTANA, P.; LAHA, T.; GASSER, R.B.A portrait of the transcriptome of the neglected trematode, Fasciola gigantica-biological and biotechnological implications. Trop. Dis. 5(2): e1004. 2011. https://doi.org/dj4ff5.