Activity of selected hydrolases in excretion-secretion products and homogenates from L3 and L4 larvae of Anisakis simplex [Nematoda: Anisakidae] parasitising herring

• Autorzy: Dziekonska-Rynko J , Rokicki J. , Jablonowski Z.
• Języki publikacji: EN

Abstrakty

EN

Background. Proteolytic enzymes may serve multiple functions: they may inhibit the host′s blood clotting, protect the parasite from the host ′s immune response, facilitate parasite′s migration within a tissue by decomposing the tissue barrier, enhance the hatching and moulting of larvae, and play an important role in their feeding. Learning their identity leads to a better understanding of a host-parasite system. The objective of this study was to check, using biochemical methods, if, in addition to proteases, ES products and extracts of 3rd and 4th larval stages of Anisakis simplex (Rudolphi, 1809) contain other hydrolases. Materials and Methods. Stage-3 larvae (L 3 ) of A.simplex were removed from Baltic herring, Clupea harengus membras Linnaeus, 1761 caught in the Baltic Sea. Stage-4 larvae (L 4) were obtained from an L 3 stage culture kept in Eagle ′s medium. The solutions containing ES products were collected and dialysed at 4°C against distilled water for 24 h. Larval extracts were obtained by homogenising the larvae in a physiological saline (0.9 % NaCl) solution in a glass homogeniser. The homogenate was centrifuged for 10 min at 3000 G. The supernatant was used in enzyme activity assays. Enzymatic activity of ES products and homogenates of L 3 and L 4 larvae of A. simplex was determined with the API ZYM test. Results. The excretion-secretion product of L 3 and L 4 larvae of A. simplex revealed activities of 10-and 11 hydrolases, respectively. Activity of esterase, esterase lipase, valine arylamidase, and N-acetyl-ß-glucosaminidase in the L 4 larvae extracts was higher than the activity of a corresponding enzyme assayed in the L 3 extracts. Only in the case of acid phosphatase, its activity in L 3 ES products was twice that of the activity found in ES products of L 4 larvae. Enzymes such as trypsin, chymotrypsin, and ß-glucosidase were not detected in extracts from L 3 larvae. Conclusion. Activities of most hydrolases in the L 4 extracts were higher than the activities of corresponding enzymes assayed in the L 3 extracts. The high activity of these enzymes found in L 4 larval extracts could be related to a different feeding mechanism of stage-4 larvae.

Słowa kluczowe

EN

parasite; Baltic herring; Clupea harengus membras; herring; Nematoda; larva; Anisakidae; Anisakis simplex; excretion-secretion product

• Wydawca: -
• Czasopismo: Acta Ichthyologica et Piscatoria
• Rocznik: 2003
• Tom: 33
• Numer: 2
• Opis fizyczny: p.125-135,ref.

Twórcy

autor

Dziekonska-Rynko J

• University of Warmia and Mazury in Olsztyn, Olsztyn, Poland

autor

Rokicki J.

autor

Jablonowski Z.

Bibliografia

• Arme C., 1966. Histochemical and biochemical studies on some enzymes of Ligula intestinalis (Cestoda: Pseudophyllidea). Journal of Parasitology 52: 63–68.
• Auriault C., Ouassi M.A., Torpier G., Eisen H., Capron A., 1981. Proteolytic cleavage of IgG bound to the Fc receptor of Schistosoma mansoni schistosomula.Parasite Immunology 3: 33–44.
• van den Bossche H., Borgers M., 1973. Subcellular distribution of digestive enzymes in Ascaris suum intestine. International Journal for Parasitology 3: 59–65.
• Bradford M.M., 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding.Analytical Biochemistry 72: 248–254.
• Buczek A., Madoń T., 1998. The role salivary glands in tick feeding (Acari: Ixodida).Wiadomości Parazytologiczne 44: 653–662.
• Dziekońska-Rynko J., Rokicki J., Jabłonowski Z., Białowąs K., 2001. Influence of the pH of cultivation medium on survival and development of stage III larvae of Anisakis simplex. Wiadomości Parazytologiczne 47: 317–322.
• Grabda J., 1976. Studies on the life cycle and morphogenesis of Anisakis simplex (Rudolphi, 1809) (Nematoda: Anisakidae) cultured in vitro. Acta Ichthyologica et Piscatoria 6: 119–141.
• Hinck L.W., Ivey M.H., 1976. Proteinase activity in Ascaris suum eggs, hatching fluid and excretions-secretions. Journal of Parasitology 62: 771–774.
• Hotez P.J., Cappello M., Hawdon J., Beckers C., Sakanari J., 1994.Hyaluronidases of the gastrointestinal invasive nematodes Ancylostoma caninum and Anisakis simplex: possible functions in the pathogenesis of human zoonoses.Journal of Infectious Diseases 170: 918–926.
• Kennedy M.W., Tierney J., Ye P., McMonagle F.A., McIntosh A., McLaughlin D.,Smith J.W., 1988. The secreted and somatic antigens of the third stage larva of Anisakis simplex, and antigenic relationship with Ascaris suum, Ascaris lumbricoides,and Toxocara canis. Molecular and Biochemical Parasitology 31: 35–46.
• Kenyon F., Knox D., 2002. The proteinases of Psoroptes ovis, the sheep scab mite-their diversity and substrate specificity. Veterinary Parasitology 105: 317–325.
• Kinsella J.L., Lichtenfels J.R., Ryan M.F., 2002. A preliminary analysis of proteolytic activity of excretory-secretory products from Cyathostominea.Veterinary Parasitology 107: 73–83.
• Knox D.P., Kennedy M.W., 1988. Proteinases released by the parasitic larval stages of Ascaris suum, and their inhibition by antibody. Molecular and Biochemical Parasitology 28: 207–216.
• Kotomski G., Wędrychowicz H., 2001. Characterization of the proteinases released by infective larvae of Uncinaria stenocephala during in vitro exsheathment and incubation. Helminthologia 38: 57–61.
• Lee D.L., 1962. The histochemical localization of leucine aminopeptidase in Ascaris lumbricoides. Parasitology 52: 533–538.
• Lee D.L., 1996. Why do some nematode parasites of the alimentary tract secrete acetylcholinesterase? International Journal for Parasitology 26: 499–508.
• Lee D.L., Foster N., 1995. Gastrointestinal nematodes and host gut motility. Helminthologia 31: 107–110.
• Łopieńska E., Żółtowska K., Rokicki J., 2001. Porównanie własności a-amylazy z larw trzeciego i czwartego stadium Anisakis simplex. [The comparison of properties of a-amylase from the third and fourth stage of Anisakis simplex larvae.]Wiadomości Parazytologiczne 47: 323–327. (In Polish.)
• Maki J., Yanagisawa T., 1980. Acid phosphatase activity demonstrated in the nematodes, Dirofilaria immitis and Angiostrongylus cantoniensis with special reference to the characters and distribution. Parasitology 80: 23–38.
• Matthews B., 1982. Behaviour and enzyme release by Anisakis sp. larvae (Nematoda: Ascaridida). Journal of Helminthology 56: 177–183.
• Matthews B., 1984. The source, release and specificity of proteolytic enzyme activity produced Anisakis simplex larvae (Nematoda: Ascaridida) in vitro. Journal of Helminthology 58: 175–185.
• McKerrow J.H., 1989. Parasite Proteases. Experimental Parasitology 68: 111–115.
• Moczoń T., 1999. A metalloproteinase secreted by the infective larvae of Strongyloides papillosus (Nematoda). Acta Parasitologica 44: 193–198.
• Moczoń T., Wranicz M., 1999. Trichinella spiralis: proteinases in the larvae. Parasitology Research 85: 47–58.
• Morris S.R., Sakanari J. A., 1994. Characterization of the serine protease and serie protease inhibitor from the tissue-penetrating nematode Anisakis simplex. Journal of Biological Chemistry 269: 27650–27656.
• Niemczuk W., 1993. Lokalizacja i aktywność wybranych enzym? w tkankach tasiemc? Bothriocephalus acheilognathi i Khawia sinensis pasożytujących u karpi. [Localization and activity of several enzymes in tissues of Bothriocephalus acheilognathi and Khawia sinensis tapeworms parasitizing carp.]Wiadomości Parazytologiczne 39: 189–197. (In Polish.)
• Nisbet A.J., Billingsley P.F., 1999. Hydrolytic enzymes of Psoroptes cuniculi (Delafond). Insect Biochemistry and Molecular Biology 29: 25–32.
• Nisbet A.J., Billingsley P.F. 2000. Acomparative survey of the hydrolytic enzymes of ectoparasitic and free-living mites. International Journal for Parasitology 30: 19–27.
• Nisbet A.J., Billingsley P.F., 2002. Characterisation of aminopeptidase activity In scab mites (Psoroptes spp.). Insect Biochemistry and Molecular Biology 32:1123–1131.
• Opperman C.H., Chang S., 1992. Nematode acetylcholinesterases: molecular forms and their potential role in nematode behavior. Parasitology Today 8: 406–411.
• Perteguer M.J., Raposo R., Cuellar C., 1996. In vitro study on the effect of larval excretory/secretory products and crude extracts from Anisakis simplex on blond coagulation. International Journal for Parasitology 26: 105–108.
• Rhoads M.L., Fetterer R.H., 1998. Purification and characterisation of a secreted aminopeptidase from adult Ascaris suum. International Journal for Parasitology 28: 1681–1690.
• Rhoads M.L., Fetterer R.H., Urban J.F. Jr., 1997. Secretion of an aminopeptidase during transition of third- to fourth-stage larvae of Ascaris suum. Journal of Parasitology 83: 780–784.
• Rhoads M.L., Fetterer R.H., Urban J.F. Jr., 1998. Effect of protease class-specific inhibitors on in vitro development of third- to fourth-stage larvae of Ascaris suum.Journal of Parasitology 84: 686–690.
• Rhodes M.B., Marsh C.L., Ferguson D.L., 1966. Studies in helminth enzymology.V. An aminopeptidase of Ascaris suum which hydrolyzes L-leucyl-b--naphthylamide. Experimental Parasitology 19: 42–51.
• Rhodes M.B., Marsh C.L., Ferguson D.L., 1969a. Studies in helminth enzymology.VI. Aminopeptidases from uterine extracts of Ascaris suum. Experimental Parasitology 26: 140–149.
• Rhodes M.B., Marsh C.L., Ferguson D.L., 1969b. Ascaris suum: Purification and characterization of an intestinal aminopeptidase. Experimental Parasitology 26: 150–155.
• Riga E., Perry R.N., Barrett J., Johnston M.R.L., 1995. Biochemical analyses on single amphidial glands, excretory-secretory gland cells, pharyngeal glands, and their secretions from the avian nematode Syngamus trachea. International Journal for Parasitology 25: 1151–1158.
• Rogers W.P., 1958. Physiology of the hatching of eggs of Ascaris lumbricoides.Nature 181: 1410–1411.
• Rogers W.P., 1963. Physiology of infection with nematodes: some effects of the host stimulus on infective stages. Annals of New York Academy of Sciences 113:208–216.
• Rogers W.P., 1982. Enzymes in the exsheathing fluid of nematodes and their biological significance. International Journal for Parasitology 12: 495–502.
• Ruitenberg E.J., Loendersloot H.J., 1971a. Histochemical properties of the excretory organ of Anisakis sp. larva. Journal of Parasitology 57: 1149–1150.
• Ruitenberg E.J., Loendersloot H.J., 1971b. Enzymhistochemische onderzoek van Anisakis sp. larven. Tidjschrift voor Diergeneeskunde 96: 247–260.
• Sajid M., McKerrow J.H., 2002. Cysteine proteases of parasitic organisms.Molecular and Biochemical Parasitology 120: 1–21.
• Sakanari J.A., McKerrow J.H., 1990. Identification of the secreted neutral proteases from Anisakis simplex. Journal of Parasitology 76: 625–630.
• Skotarczak B., 1987. Badanie histochemiczne jaj Ascaris suum w czasie embriogenezy. I. Fosfatazy. [Histochemical studies on eggs of Ascaris suum during embryogenesis. I. Phosphatases.] WiadomoEci Parazytologiczne 33: 681–686.(In Polish.)
• Sommerville R.I., Davey K.G., 1976. Stimuli for cuticle formation and ecdysis In vitro of the infective larvae of Anisakis sp. (Nematoda: Ascaridoidea).International Journal for Parasitology 6: 433–439.
• Xu Y., Dresden M.H., 1986. Leucine aminopeptidase and hatching of Schistosoma mansoni eggs. Journal of Parasitology 72: 507–511.
• Young A.R., Mancuso N., Bowles V.M., 1999. Biochemical aspects of egg hatch In endo- and ectoparasites: potential for rational drug design. International Journal for Parasitology 29: 861–867.
• Żółtowska K., Łopieńska E., Rokicki J., 2000. Content of carbohydrates and activity of sugar metabolism enzymes present in stage III larvae of Anisakis simplex. Acta Parasitologica 45: 186.