{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,20]],"date-time":"2026-09-20T03:27:01Z","timestamp":1789874821950,"version":"4.0.1"},"reference-count":35,"publisher":"Cambridge University Press (CUP)","issue":"3","license":[{"start":{"date-parts":[[2001,11,28]],"date-time":"2001-11-28T00:00:00Z","timestamp":1006905600000},"content-version":"unspecified","delay-in-days":272,"URL":"https:\/\/www.cambridge.org\/core\/terms"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Parasitology"],"published-print":{"date-parts":[[2001,3]]},"abstract":"<jats:p>This paper describes the<jats:italic>in vitro<\/jats:italic>cultivation of the 3rd-larval stage (L3) of<jats:italic>Anisakis simplex<\/jats:italic>to adulthood in a much simpler and easier to prepare medium than those described to date. The adult males obtained are between 3.8 and 6.5\u00a0cm long and the females between 4.5 and 8.0\u00a0cm. Some individually cultivated females laid eggs which had an average size of 44.4\u00d750.5\u00a0\u03bcm. The culture conditions were as follows: medium RPMI-1640\u00a0supplemented with 20% heat-inactivated fetal bovine serum and 1% commercial pepsin, at pH\u00a04.0 and a temperature of 37\u00a0\u00b0C, and in air atmosphere with 5% CO<jats:sub>2<\/jats:sub>. The pepsin was found to be the key to the success of the culture. The average survival of the worms in the culture increased from 50 to 88 days, due to the fact that the survival of the adults practically doubled (increasing by 1.9 times). Furthermore, the number of worms that completed the 4th moulting (M4) increased by 4.2 times, from 22.9 to 95.6%. This culture medium may facilitate, due to its simplicity, the study of anisakids, or at least of<jats:italic>A. simplex,<\/jats:italic>constituting another step towards achieving a complete<jats:italic>in vitro<\/jats:italic>life-cycle for these parasites.<\/jats:p>","DOI":"10.1017\/s0031182001008423","type":"journal-article","created":{"date-parts":[[2002,10,30]],"date-time":"2002-10-30T15:59:07Z","timestamp":1035993547000},"page":"285-291","source":"Crossref","is-referenced-by-count":56,"title":["<i>In vitro<\/i>cultivation of<i>Anisakis simplex:<\/i>pepsin increases survival and moulting from fourth larval to adult stage"],"prefix":"10.1017","volume":"123","author":[{"given":"L.","family":"IGLESIAS","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A.","family":"VALERO","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R.","family":"BEN\u00cdTEZ","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"F.\u00a0J.","family":"ADROHER","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"56","published-online":{"date-parts":[[2001,11,28]]},"reference":[{"key":"S0031182001008423_ref027","doi-asserted-by":"publisher","DOI":"10.1016\/0169-4758(90)90176-5"},{"key":"S0031182001008423_ref024","doi-asserted-by":"publisher","DOI":"10.2307\/3284267"},{"key":"S0031182001008423_ref035","doi-asserted-by":"publisher","DOI":"10.1080\/00364827.1999.10807355"},{"key":"S0031182001008423_ref018","doi-asserted-by":"publisher","DOI":"10.1016\/0020-7519(86)90032-9"},{"key":"S0031182001008423_ref017","unstructured":"MIKKELSEN, B. (1998). Summer diet of grey seals Halichoerus grypus in the Faroe Islands. Thesis. Department of Aquatic Resources and Environmental Biology. Norwegian College of Fishery Science, University of Troms\u00f8, Norway."},{"key":"S0031182001008423_ref019","doi-asserted-by":"crossref","unstructured":"OLSEN, M.\u00a0A. , AAGNES, T.\u00a0H. & MATHIESEN, S.\u00a0D. (1994a). Digestion of herring by indigenous bacteria in the minke whale forestomach. Applied and Environmental Microbiology 60, 4445\u20134455.","DOI":"10.1128\/AEM.60.12.4445-4455.1994"},{"key":"S0031182001008423_ref007","unstructured":"IGLESIAS, L. , VALERO, A. & ADROHER, F.\u00a0J. (1997). Some factors which influence the in vitro maintenance of Anisakis simplex (Nematoda). Folia Parasitologica 44, 297\u2013301."},{"key":"S0031182001008423_ref013","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.47.30181"},{"key":"S0031182001008423_ref026","unstructured":"RUIZ-VALERO, J. , VALERO, A. , ADROHER, F.\u00a0J. & ORTEGA, J.\u00a0E. (1992). Presencia de asc\u00e1ridos en peces comerciales de frecuente consumo en Granada. In In Memorian al Profesor Doctor D.\u00a0F. de P. Mart\u00ednez G\u00f3mez (ed. HERN\u00c1NDEZ, S .), pp. 335\u2013349. C\u00f3rdoba, Spain, Universidad de C\u00f3rdoba."},{"key":"S0031182001008423_ref009","doi-asserted-by":"publisher","DOI":"10.1038\/227680a0"},{"key":"S0031182001008423_ref030","unstructured":"SOMMERVILLE, R.\u00a0I. & DAVEY, K.\u00a0G. (1976). Stimuli for cuticle formation and ecdysis in vitro of the infective larva of Anisakis sp. (Nematoda: Ascaridoidea). International Journal for Parasitology 6, 433\u2013439."},{"key":"S0031182001008423_ref033","doi-asserted-by":"crossref","unstructured":"VAN BANNING, P. (1971). Some notes on a successful rearing of the herring-worm, Anisakis marina L. (Nematoda: Heterocheilidae). Journal du Conseil International de l'Exploration de la Mer 34, 84\u201388.","DOI":"10.1093\/icesjms\/34.1.84"},{"key":"S0031182001008423_ref010","doi-asserted-by":"publisher","DOI":"10.1139\/f89-141"},{"key":"S0031182001008423_ref008","unstructured":"KIKUCHI, Y. , UEDA, T. , YOSHIKI, T. , AIZAWA, M. & ISHIKURA, H. (1967). Experimental studies of the immunopathology of the intestinal anisakiasis. Igakuno Ayumi 62, 731\u2013736 (in Japanese)."},{"key":"S0031182001008423_ref004","unstructured":"DZIEKO\u0143SKA-RYNKO, J. , ROKICKI, J. & JAB\u0142ONOWSKI, Z. (1997). The impact of animal and plant proteases on Nematoda 3rd stage Anisakis simplex larvae. Oceanological Studies 1, 103\u2013107."},{"key":"S0031182001008423_ref023","unstructured":"PODOLSKA, M. , PIUSI\u0143SKI, W. & ROKICKI, J. (1997). 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