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

Molecular and Cellular Proteomics

Volumn 8, Issue 8, 2009, Pages 1891-1907

  Robinson, Mark W ^a   Menon, Ranjeeta ^b   Donnelly, Sheila M ^a   Dalton, John P ^a   Ranganathan, Shoba ^b  

^a UNIVERSITY OF TECHNOLOGY SYDNEY   (Australia)

^b MACQUARIE UNIVERSITY   (Australia)

Author keywords

[No Author keywords available]

Indexed keywords

ANTIOXIDANT; ASPARAGINYL ENDOPEPTIDASE CYSTEINE PROTEASE; CARBOXYPEPTIDASE; CATHEPSIN B; CATHEPSIN L; CYSTEINE PROTEINASE; PROTEASE FHCL1; PROTEASE FHCL2; PROTEASE FHCL3; PROTEASE FHCL5; PROTEINASE; SECRETORY PROTEIN; SERINE PROTEINASE; TRYPSIN LIKE SERINE PROTEASE; UNCLASSIFIED DRUG;

ANIMAL EXPERIMENT; ANIMAL TISSUE; ARTICLE; AUTOMATION; BLOOD; CONTROLLED STUDY; EXPRESSED SEQUENCE TAG; FASCIOLA HEPATICA; FEEDING; FEMALE; HELMINTH; HOST; HOST PATHOGEN INTERACTION; INTESTINE WALL; LARVA; LIVER; MACROMOLECULE; MAMMAL; MOUSE; NONHUMAN; NUCLEOTIDE SEQUENCE; PATHOGENESIS; PRIORITY JOURNAL; PROTEIN ANALYSIS; PROTEIN EXPRESSION; PROTEIN FUNCTION; PROTEIN SECRETION; PROTEOMICS; SIGNAL TRANSDUCTION; TISSUE; TRANSCRIPTOMICS; TREMATODE;

ANIMALS; ELECTROPHORESIS, POLYACRYLAMIDE GEL; EXPRESSED SEQUENCE TAGS; FASCIOLA HEPATICA; FASCIOLIASIS; GENE EXPRESSION PROFILING; HELMINTH PROTEINS; HUMANS; MASS SPECTROMETRY; PROTEOMICS; VIRULENCE;

FASCIOLA; FASCIOLA HEPATICA; MAMMALIA; VERMES;

EID: 68649087906     PISSN: 15359476     EISSN: None     Source Type: Journal    
DOI: 10.1074/mcp.M900045-MCP200     Document Type: Article

References (83)

1. Garcia, H. H., Moro, P. L., and Schantz, P. M. (2007) Zoonotic helminth infections of humans: echinococcosis, cysticercosis and fascioliasis. Curr. Opin. Infect. Dis. 20, 489-494
2. Andrews, S. J. (1999) The life-cycle of Fasciola hepatica, in Fasciolosis (Dalton, J. P., ed) pp. 1-29, CAB International, Oxford
3. Mas-Coma, S., Bargues, M. D., and Esteban, J. G. (1999) Human fasciolosis, in Fasciolosis (Dalton, J. P., ed) pp. 411-434, CAB International, Oxford
4. Fairweather, I., Threadgold, L. T., and Hanna, R. E. (1999) Development of Fasciola hepatica in the mammalian host, in Fasciolosis (Dalton, J. P., ed) pp. 47-111, CAB International, Oxford
5. Hewitson, J. P., Harcus, Y. M., Curwen, R. S., Dowle, A. A., Atmadja, A. K., Ashton, P. D., Wilson, A., and Maizels, R. M. (2008) The secretome of the filarial parasite, Brugia malayi: proteomic profile of adult excretory-secretory products. Mol. Biochem. Parasitol. 160, 8-21
6. Robinson, M. W., and Connolly, B. (2005) Proteomic analysis of the excretory-secretory proteins of the Trichinella spiralis L1 larva, a nematode parasite of skeletal muscle. Proteomics 5, 4525-4532
7. Robinson, M. W, Greig, R., Beattie, K. A., Lamont, D. J., and Connolly, B. (2007) Comparative analysis of the excretory-secretory proteome of the muscle larva of Trichinella pseudospiralis and Trichinella spiralis. Int. J. Parasitol. 37, 139-148
8. Robinson, M. W., Tort, J. F., Lowther, J., Donnelly, S. M., Wong, E., Xu, W., Stack, C. M., Padula, M., Herbert, B., and Dalton, J. P. (2008) Proteomics and phylogenetic analysis of the cathepsin L protease family of the helminth pathogen, Fasciola hepatica: expansion of a repertoire of virulence-associated factors. Mol. Cell. Proteomics 7, 1111-1123
9. Knudsen, G. M., Medzihradszky, K. F., Lim, K. C., Hansell, E., and McKerrow, J. H. (2005) Proteomic analysis of Schistosoma mansoni cercarial secretions. Mol. Cell. Proteomics 4, 1862-1875
10. Ojopi, E. P., Oliveira, P. S., Nunes, D. N., Paquola, A., DeMarco, R., Gregório, S. P., Aires, K. A., Menck, C. F., Leite, L. C., Verjovski-Almeida, S., and Dias-Neto, E. (2007) A quantitative view of the transcriptome of Schistosoma mansoni adult-worms using SAGE. BMC Genomics 8, 186 (Pubitemid 47189795)
11. Oliveira, G. (2007) The Schistosoma mansoni transcriptome: an update. Exp. Parasitol. 117, 229-235
12. Laha, T., Pinlaor, P., Mulvenna, J., Sripa, B., Sripa, M., Smout, M. J., Gasser, R. B., Brindley, P. J., and Loukas, A. (2007) Gene discovery for the carcinogenic human liver fluke, Opisthorchis viverrini. BMC Genomics 8, 189
13. Nagaraj, S. H., Gasser, R. B., and Ranganathan, S. (2008) Needles in the EST haystack: large-scale identification and analysis of excretory-secretory (ES) proteins in parasitic nematodes using expressed sequence tags (ESTs). PLoS Negl. Trop. Dis. 2, e301
14. Nagaraj, S. H., Deshpande, N., Gasser, R. B., and Ranganathan, S. (2007) ESTExplorer: an expressed sequence tag (EST) assembly and annotation platform. Nucleic Acids Res. 35, W143-147
15. Nagaraj, S. H., Gasser, R. B., Nisbet, A. J., and Ranganathan, S. (2008) In silico analysis of expressed sequence tags from Trichostrongylus vitrinus (Nematoda): comparison of the automated ESTExplorer workflow platform with conventional database searches. BMC Bioinformatics 9, Suppl. 1, S10
16. Ranganathan, S., Nagaraj, S. H., Hu, M., Strube, C., Schnieder, T., and Gasser, R. B. (2007) A transcriptomic analysis of the adult stage of the bovine lungworm, Dictyocaulus viviparus. BMC Genomics 8, 311
17. Datu, B. J., Gasser, R. B., Nagaraj, S. H., Ong, E. K., O'Donoghue, P., McInnes, R., Ranganathan, S., and Loukas, A. (2008) Transcriptional changes in the hookworm, Ancylostoma caninum, during the transition from a free-living to a parasitic larva. PLoS Negl. Trop. Dis. 2, e130
18. Altschul, S. F., Madden, T. L., Schärfer, A. A., Zhang, J., Zhang, Z., Miller, W., and Lipman, D. J. (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 25, 3389-3402 (Pubitemid 27359211)
19. Zdobnov, E. M, and Apweiler, R. (2001) InterProScan: an integration platform for the signature-recognition methods in InterPro. Bioinformatics 17, 847-848
20. Jefferies, J. R., Campbell, A. M., van Rossum, A. J., Barrett, J., and Brophy, P. M. (2001) Proteomic analysis of Fasciola hepatica excretory-secretory products. Proteomics 1, 1128-1132 (Pubitemid 33696478)
21. Morphew, R. M., Wright, H. A., LaCourse, E. J., Woods, D. J., and Brophy, P. M. (2007) Comparative proteomics of excretory-secretory proteins released by the liver fluke Fasciola hepatica in sheep host bile and during in vitro culture ex host. Mol. Cell. Proteomics 6, 963-972
22. Ishihama, Y., Oda, Y., Tabata, T., Sato, T., Nagasu, T., Rappsilber, J., and Mann, M. (2005) Exponentially modified protein abundance index (emPAI) for estimation of absolute protein amount in proteomics by the number of sequenced peptides per protein. Mol. Cell. Proteomics 4, 1265-1272
23. El-Sayed, N. M., Bartholomeu, D., Ivens, A., Johnston, D. A., and LoVerde, P. T. (2004) Advances in schistosome genomics. Trends Parasitol. 20, 154-157
24. Mitreva, M., Zarlenga, D. S., McCarter, J. P., and Jasmer, D. P. (2007) Parasitic nematodes: from genomes to control. Vet. Parasitol. 148, 31-42
25. Stein, L. D., Bao, Z., Blasiar, D., Blumenthal, T., Brent, M. R., Chen, N., Chinwalla, A., Clarke, L., Clee, C., Coghlan, A., Coulson, A., D'Eustachio, P., Fitch, D. H., Fulton, L. A., Fulton, R. E., Griffiths-Jones, S., Harris, T. W., Hillier, L. W., Kamath, R., Kuwabara, P. E., Mardis, E. R., Marra, M. A., Miner, T. L., Minx, P., Mullikin, J. C., Plumb, R. W., Rogers, J., Schein, J. E., Sohrmann, M., Spieth, J., Stajich, J. E., Wei, C., Willey, D., Wilson, R. K., Durbin, R., and Waterston, R. H. (2003) The genome sequence of Caenorhabditis briggsae: a platform for comparative genomics. PLoS Biol. 1, E45 (Pubitemid 39112161)
26. Chen, Y., Zhang, Y., Yin, Y., Gao, G., Li, S., Jiang, Y., Gu, X., and Luo, J. (2005) SPD - a web-based secreted protein database. Nucleic Acids Res. 33, D169-173 (Pubitemid 40207853)
27. Choo, K. H., Tan, T. W., and Ranganathan, S. (2005) SPdb - a signal peptide database. BMC Bioinformatics 6, 249
28. Cancela, M., Acosta, D., Rinaldi, G., Silva, E., Durán, R., Roche, L., Zaha, A., Carmona, C., and Tort, J. F. (2008) A distinctive repertoire of cathepsins is expressed by juvenile invasive Fasciola hepatica. Biochimie 90, 1461-1475
29. Grams, R., Adisakwattana, P., Ritthisunthorn, N., Eursitthichai, V., Vichasri-Grams, S., and Viyanant, V. (2006) The saposin-like proteins 1, 2, and 3 of Fasciola gigantica. Mol. Biochem. Parasitol. 148, 133-143
30. Espino, A. M., and Hillyer, G. V. (2003) Molecular cloning of a member of the Fasciola hepatica saposin-like protein family. J. Parasitol. 89, 545-552 (Pubitemid 36828526)
31. Robinson, M. W., Colhoun, L. M., Fairweather, I., Brennan, G. P., and Waite, J. H. (2001) Development of the vitellaria of the liver fluke, Fasciola hepatica in the rat host. Parasitology 123, 509-518
32. Salazar-Calderón, M., Martín-Alonso, J. M., Castro, A. M., and Parra, F. (2003) Cloning, heterologous expression in Escherichia coli and characterization of a protein disulfide isomerase from Fasciola hepatica. Mol. Biochem. Parasitol. 126, 15-23
33. Bork, P., and Beckmann, G. (1993) The CUB domain. A widespread module in developmentally regulated proteins. J. Mol. Biol. 231, 539-545
34. Tkalcevic, J., Ashman, K., and Meeusen, E. (1995) Fasciola hepatica: rapid identification of newly excysted juvenile proteins. Biochem. Biophys. Res. Commun. 213, 169-174
35. Irving, J. A., Spithill, T. W., Pike, R. N., Whisstock, J. C., and Smooker, P. M. (2003) The evolution of enzyme specificity in Fasciola spp. J. Mol. Evol. 57, 1-15
36. Lowther, J., Robinson, M. W., Donnelly, S. M., Xu, W., Stack, C. M., Matthews, J. M., and Dalton, J. P. (2009) The importance of pH in regulating the function of Fasciola hepatica cathepsin L1 cysteine protease. PLoS. Negl. Trop. Dis. 3, e369
37. Dalton, J. P., Caffrey, C. R., Sajid, M., Stack, C., Donnelly, S., Loukas, A., Don, T., McKerrow, J., Halton, D. W., and Brindley, P. J. (2006) Proteases in trematode biology, in Parasitic Flatworms: Molecular Biology, Biochemistry, Immunology and Physiology (Maule, A. G., and Marks, N. J., eds) pp. 348-368, CAB International, Oxford
38. Stack, C. M., Caffrey, C. R., Donnelly, S. M., Seshaadri, A., Lowther, J., Tort, J. F., Collins, P. R., Robinson, M. W., Xu, W., McKerrow, J. H., Craik, C. S., Geiger, S. R., Marion, R., Brinen, L. S., and Dalton, J. P. (2008) Structural and functional relationships in the virulence-associated cathepsin L proteases of the parasitic liver fluke, Fasciola hepatica. J. Biol. Chem. 283, 9896-9908
39. Don, T. A., Oksov, Y., Lustigman, S., and Loukas, A. (2007) Saposin-like proteins from the intestine of the blood-feeding hookworm, Ancylostoma caninum. Parasitology 134, 427-436
40. Law, R. H., Smooker, P. M., Irving, J. A., Piedrafita, D., Ponting, R., Kennedy, N. J., Whisstock, J. C., Pike, R. N., and Spithill, T. W. (2003) Cloning and expression of the major secreted cathepsin B-like protein from juvenile Fasciola hepatica and analysis of immunogenicity following liver fluke infection. Infect. Immun. 71, 6921-6932
41. Adisakwattana, P., Viyanant, V., Chaicumpa, W., Vichasri-Grams, S., Hofmann, A., Korge, G., Sobhon, P., and Grams, R. (2007) Comparative molecular analysis of two asparaginyl endopeptidases and encoding genes from Fasciola gigantica. Mol. Biochem. Parasitol. 156, 102-116
42. Turk, D., and Guncar, G. (2003) Lysosomal cysteine proteases (cathepsins): promising drug targets. Acta Crystallogr. D Biol. Crystallogr. 59, 203-213
43. Chen, J. M., Dando, P. M., Stevens, R. A., Fortunato, M., and Barrett, A. J. (1998) Cloning and expression of mouse legumain, a lysosomal endopeptidase. Biochem. J. 335, 111-117 (Pubitemid 28477056)
44. Michel, A., Ghoneim, H., Resto, M., Klinkert, M. Q., and Kunz, W. (1995) Sequence, characterization and localization of a cysteine proteinase cathepsin L in Schistosoma mansoni. Mol. Biochem. Parasitol. 73, 7-18
45. Meemon, K., Grams, R., Vichasri-Grams, S., Hofmann, A., Korge, G., Viyanant, V., Upatham, E. S., Habe, S., and Sobhon, P. (2004) Molecular cloning and analysis of stage and tissue-specific expression of cathepsin B encoding genes from Fasciola gigantica. Mol. Biochem. Parasitol. 136, 1-10
46. Gourbal, B. E., Guillou, F., Mitta, G., Sibille, P., Thèron, A., Pointier, J. P., and Coustau, C. (2008) Excretory-secretory products of larval Fasciola hepatica investigated using a two-dimensional proteomic approach. Mol. Biochem. Parasitol. 161, 63-66
47. Salazar-Calderón, M., Martin-Alonso, J. M., Ruiz de Eguino, A. D., and Parra, F. (2001) Heterologous expression and functional characterization of thioredoxin from Fasciola hepatica. Parasitol Res. 87, 390-395 (Pubitemid 32391441)
48. Sekiya, M., Mulcahy, G., Irwin, J. A., Stack, C. M., Donnelly, S. M., Xu, W., Collins, P., and Dalton, J. P. (2006) Biochemical characterisation of the recombinant peroxiredoxin (FhePrx) of the liver fluke, Fasciola hepatica. FEBS Lett. 580, 5016-5022
49. Emanuelsson, O., Brunak, S., von Heijne, G., and Nielsen, H. (2007) Locating proteins in the cell using TargetP, SignalP, and related tools. Nat. Protoc. 2, 953-971
50. Dalton, J. P., Skelly, P., and Halton, D. W. (2004) Role of the tegument and gut in nutrient uptake by parasitic platyhelminths. Can. J. Zool. 82, 211-232
51. Keller, M., Rüegg, A., Werner, S., and Beer, H. D. (2008) Active caspase-1 is a regulator of unconventional protein secretion. Cell 132, 818-831 (Pubitemid 351312798)
52. MacKenzie, A., Wilson, H. L., Kiss-Toth, E., Dower, S. K., North, R. A., and Surprenant, A. (2001) Rapid secretion of interieukin-1beta by microvesicle shedding. Immunity 15, 825-835 (Pubitemid 34008509)
53. Andrei, C., Margiocco, P., Poggi, A., Lotti, L. V., Torrisi, M. R., and Rubartelli, A. (2004) Phospholipases C and A2 control lysosome-mediated IL-1 beta secretion: implications for inflammatory processes. Proc. Natl. Acad. Sci. U.S.A. 101, 9745-9750
54. Mambula, S. S., Stevenson, M. A., Ogawa, K., and Calderwood, S. K. (2007) Mechanisms for Hsp70 secretion: crossing membranes without a leader. Methods 43, 168-175 (Pubitemid 47499072)
55. Kumkate, S., Chunchob, S., and Janvilisri, T. (2008) Expression of ATP-binding cassette multidrug transporters in the giant liver fluke Fasciola gigantica and their possible involvement in the transport of bile salts and anthelmintics. Mol. Cell. Biochem. 317, 77-84
56. Harmsen, M. M., Cornelissen, J. B., Buijs, H. E., Boersma, W. J., Jeurissen, S. H., and van Milligen, F. J. (2004) Identification of a novel Fasciola hepatica cathepsin L protease containing protective epitopes within the propeptide. Int. J. Parasitol. 34, 675-682
57. Beckham, S. A., Law, R. H., Smooker, P. M., Quinsey, N. S., Caffrey, C. R., McKerrow, J. H., Pike, R. N., and Spithill, T. W. (2006) Production and processing of a recombinant Fasciola hepatica cathepsin B-like enzyme (FhcatB1) reveals potential processing mechanisms in the parasite. Biol. Chem. 387, 1053-1061
58. Coulombe, R., Grochulski, P., Sivaraman, J., Ménard, R., Mort, J. S., and Cygler, M. (1996) Structure of human procathepsin L reveals the molecular basis of inhibition by the prosegment. EMBO J. 15, 5492-5503 (Pubitemid 26385609)
59. Dalton, J. P., Brindley, P. J., Donnelly, S., and Robinson, M. W. (2009) The enigmatic asparaginyl endopeptidase of helminth parasites. Trends Parasitol. 25, 59-61
60. Dalton, J. P., and Brindley, P. J. (1996) Schistosome asparaginyl endopeptidase Sm 32 in hemoglobin digestion. Parasitol. Today 12, 125
61. Robinson, M. W., Dalton, J. P., and Donnelly, S. (2008) Helminth pathogen cathepsin proteases: it's a family affair. Trends Biochem. Sci. 33, 601-608
62. Sajid, M., McKerrow, J. H., Hansell, E., Mathieu, M. A., Lucas, K. D., Hsieh, I., Greenbaum, D., Bogyo, M., Salter, J. P., Lim, K. C., Franklin, C., Kim, J. H., and Caffrey, C. R. (2003) Functional expression and characterization of Schistosoma mansoni cathepsin B and its trans-activation by an endogenous asparaginyl endopeptidase. Mol. Biochem. Parasitol. 131, 65-75
63. Chen, J. M., Fortunato, M., and Barrett, A. J. (2000) Activation of human prolegumain by cleavage at a C-terminal asparagine residue. Biochem. J. 352, 327-334 (Pubitemid 32011426)
64. Morita, Y., Araki, H., Sugimoto, T., Takeuchi, K., Yamane, T., Maeda, T., Yamamoto, Y., Nishi, K., Asano, M., Shirahama-Noda, K., Nishimura, M., Uzu, T., Hara-Nishimura, I., Koya, D., Kashiwagi, A., and Ohkubo, I. (2007) Legumain/asparaginyl endopeptidase controls extracellular matrix remodeling through the degradation of fibronectin in mouse renal proximal tubular cells. FESS Lett. 581, 1417-1424 (Pubitemid 46452617)
65. Behm, C. A., and Sangster, N. C. (1999) Pathology, pathophysiology and clinical aspects, in Fasciolosis (Dalton, J. P., ed) pp. 185-224, CAB International, Oxford
66. Geldhof, P., and Knox, D. (2008) The intestinal contortin structure in Haemonchus contortus: an immobilised anticoagulant? Int. J. Parasitol. 38, 1579-1588
67. McGonigle, L., Mousley, A., Marks, N. J., Brennan, G. P., Dalton, J. P., Spithill, T. W., Day, T. A., and Maule, A. G. (2008) The silencing of cysteine proteases in Fasciola hepatica newly excysted juveniles using RNA interference reduces gut penetration. Int. J. Parasitol. 38, 149-155
68. Kasny, M., Dalton, J. P., Mikes, L., and Horák, P. (2007) Comparison of cysteine and serine peptidase activities in Trichobilharzia regenti and Schistosoma mansoni cercariae. Parasitology 134, 1599-1609 (Pubitemid 350194872)
69. Ikeda, T. (2003) Involvement of cysteine proteinases in excystment of Paragonimus ohirai metacercariae induced by sodium cholate and A23187. J. Helminthol. 77, 21-26
70. Chung, Y. B., Kim, T. S., and Yang, H. J. (2005) Early cysteine protease activity in excretory bladder triggers metacercaria excystment of Paragonimus westermani. J. Parasitol. 91, 953-954
71. Na, B. K., Kim, S. H., Lee, E. G., Kim, T. S., Bae, Y. A., Kang, I., Yu, J. R., Sohn, W. M., Cho, S. Y., and Kong, Y. (2006) Critical roles for excretory-secretory cysteine proteases during tissue invasion of Paragonimus westermani newly excysted metacercariae. Cell. Microbiol. 8, 1034-1046
72. Smooker, P. M., Whisstock, J. C., Irving, J. A., Siyaguna, S., Spithill, T. W., and Pike, R. N. (2000) A single amino acid substitution affects substrate specificity in cysteine proteinases from Fasciola hepatica. Protein Sci. 9, 2567-2572 (Pubitemid 32105739)
73. Cervi, L., Rossi, G., and Masih, D. T. (1999) Potential role for excretory-secretory forms of glutathione-S-transferase (GST) in Fasciola hepatica. Parasitology 119, 627-633
74. Donnelly, S., O'Neill, S. M., Sekiya, M., Mulcahy, G., and Dalton, J. P. (2005) Thioredoxin peroxidase secreted by Fasciola hepatica induces the alternative activation of macrophages. Infect. Immun. 73, 166-173
75. Donnelly, S., Stack, C. M., O'Neill, S. M., Sayed, A. A., Williams, D. L., and Dalton, J. P. (2008) Helminth 2-Cys peroxiredoxin drives Th2 responses through a mechanism involving alternatively activated macrophages. FASEB J. 22, 4022-4032
76. McGonigle, S., Curley, G. P., and Dalton, J. P. (1997) Cloning of peroxiredoxin, a novel antioxidant enzyme, from the helminth parasite Fasciola hepatica. Parasitology 115, 101-104 (Pubitemid 27337233)
77. Tielens, G. M. (1999) Metabolism, in Fasciolosis (Dalton, J. P., ed) pp. 277-306, CAB International, Oxford
78. Collins, P. R., Stack, C. M., O'Neill, S. M., Doyle, S., Ryan, T., Brennan, G. P., Mousley, A., Stewart, M., Maule, A. G., Dalton, J. P., and Donnelly, S. (2004) Cathepsin L1, the major protease involved in liver fluke (Fasciola hepatica) virulence: propeptide cleavage sites and autoactivation of the zymogen secreted from gastrodermal cells. J. Biol. Chem. 279, 17038-17046
79. Robinson, M. W., Trudgett, A., Hoey, E. M., and Fairweather, I. (2002) Triclabendazole-resistant Fasciola hepatica: β-tubulin and response to in vitro treatment with triclabendazole. Parasitology 124, 325-338 (Pubitemid 34250745)
80. Meaney, M., Allister, J., McKinstry, B., McLaughlin, K., Brennan, G. P., Forbes, A. B., and Fairweather, I. (2007) Fasciola hepatica: ultrastructural effects of a combination of triclabendazole and clorsulon against mature fluke. Parasitol. Res. 100, 1091-1104
81. Meaney, M., Haughey, S., Brennan, G. P., and Fairweather, I. (2005) Ultrastructural observations on oral ingestion and trans-tegumental uptake of clorsulon by the liver fluke, Fasciola hepatica. Parasitol. Res. 95, 201-212 (Pubitemid 40379273)
82. McManus, D. P., and Dalton, J. P. (2006) Vaccines against the zoonotic trematodes Schistosoma japonicum, Fasciola hepatica and Fasciola gigantica. Parasitology 133, S43-61
83. Alcala-Canto, Y., Ibarra-Velarde, F., Sumano-Lopez, H., Gracia-Mora, J., and Alberti-Navarro, A. (2007) Effect of a cysteine protease inhibitor on Fasciola hepatica (liver fluke) fecundity, egg viability, parasite burden, and size in experimentally infected sheep. Parasitol. Res. 100, 461-465