{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T00:22:50Z","timestamp":1789690970877,"version":"4.0.1"},"reference-count":89,"publisher":"SAGE Publications","issue":"6","license":[{"start":{"date-parts":[[2003,11,1]],"date-time":"2003-11-01T00:00:00Z","timestamp":1067644800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Altern Lab Anim"],"published-print":{"date-parts":[[2003,11]]},"abstract":"<jats:p>The UK shellfish industry has recently been affected by the statutory closure of several cockle beds, following the detection of samples causing rapid and severe reactions in the regulatory approved test for diarrhetic shellfish poisoning (DSP) toxins, the mouse bioassay (MBA). It is contended that these so-called atypical results are due to procedural artefacts of the MBA; so far, several studies have failed to identify their cause. This paper critically assesses the development, regulatory use and methodological deficiencies of the MBA. It also discusses how testing for DSP toxins could and should have been improved and made more humane by applying the Three Rs concept of Reduction, Refinement and Replacement, and by the proper validation of the test method used. It is concluded that the MBA should not have been developed for the routine screening of shellfish samples, as it has a substantially severe endpoint and is not used as part of a tiered-testing strategy with non-animal methods. Moreover, during the UK monitoring programme for DSP toxins, the assay has been used without an optimised and universal protocol, and apparently without due regard to the principles of basic scientific methodology. In view of this, the atypical results obtained for cockle samples cannot be relied on as evidence of a human health hazard. It is recommended that the use of the MBA should be discontinued as soon as possible, in favour of other methods, especially those involving non-animal techniques. In the short-term, these methods should be based on analytical chemical detection systems and the essential availability of the relevant pure toxin standards. The lack of any known toxins in samples should be taken as evidence of lack of contamination. The suitability of the existing non-animal methods needs to be assessed as a matter of urgency. It is crucial that all new methods should be properly validated, and that their acceptability for their stated purposes should be endorsed by recognised criteria and validation centres, before being recommended to, or required by, regulatory agencies. In this way, the possibility that scientifically unsuitable methods will once again be used for monitoring for the contamination of shellfish with toxins can be avoided. This gross misuse of laboratory animals and ill-judged application of science should never be allowed to occur again.<\/jats:p>","DOI":"10.1177\/026119290303100608","type":"journal-article","created":{"date-parts":[[2019,4,10]],"date-time":"2019-04-10T00:28:04Z","timestamp":1554856084000},"page":"595-610","source":"Crossref","is-referenced-by-count":13,"title":["The Mouse Bioassay for Diarrhetic Shellfish Poisoning: A Gross Misuse of Laboratory Animals and of Scientific Methodology"],"prefix":"10.1177","volume":"31","author":[{"given":"Robert D.","family":"Combes","sequence":"first","affiliation":[{"name":"FRAME, Russell & Burch House, 96\u201398 North Sherwood Street, Nottingham NG1 4EE, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"179","published-online":{"date-parts":[[2003,11,1]]},"reference":[{"key":"bibr1-026119290303100608","doi-asserted-by":"publisher","DOI":"10.2216\/i0031-8884-32-2-79.1"},{"key":"bibr2-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1016\/S0924-2244(01)00006-1"},{"key":"bibr3-026119290303100608","unstructured":"Anon. (2002). Report of the ICES-IOC Working Group on Harmful Algal Bloom Dynamics, Bermuda, March 2002, ICES CM 2002\/C:03, Ref. ACME, 60 pp. Copenhagen, Denmark: International Council for the Exploration of the Sea."},{"key":"bibr4-026119290303100608","first-page":"173","volume-title":"HACCP: Hazard Analysis and Critical Control Point Training Curriculum","author":"Ward D.","year":"1997"},{"key":"bibr5-026119290303100608","unstructured":"Van DolahF.M. (2003). Excerpt from Diversity of Marine and Freshwater Algal Toxins. Website http:\/\/www.chbr.noaa.gov\/CostalResearch\/FactSheets\/Dinophysis\/DinoDiarrheticShellfishToxin.pdf (Accessed 15.10.03)."},{"key":"bibr6-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1016\/S0160-4120(01)00058-7"},{"key":"bibr7-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1016\/0006-291X(89)92189-X"},{"key":"bibr8-026119290303100608","first-page":"335","volume":"7","author":"Eriksson J.E.","year":"1993","journal-title":"Advances in Protein Phosphatases"},{"key":"bibr9-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1038\/337078a0"},{"key":"bibr10-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1016\/S0041-0101(00)00137-9"},{"key":"bibr11-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1002\/jcp.1041520124"},{"key":"bibr12-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1016\/S0887-2333(01)00021-2"},{"key":"bibr13-026119290303100608","first-page":"1","volume":"268","author":"Anon.","year":"1991","journal-title":"Official Journal of the European Communities"},{"key":"bibr14-026119290303100608","unstructured":"Anon. (1992). The Food Safety (Live Bivalve Molluscs and Other Shellfish) Regulations 1992. S.I. No. 3164. London, UK: HMSO. Website http:\/\/www.hmso.gov.uk\/si\/si1992\/UKsi_19923164_en_1.htm (Accessed 12.12.03)."},{"key":"bibr15-026119290303100608","first-page":"62","volume":"75","author":"Anon.","year":"2002","journal-title":"Official Journal of the European Communities"},{"key":"bibr16-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1016\/S0041-0101(02)00177-0"},{"key":"bibr17-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1021\/bk-1984-0262.ch019"},{"key":"bibr18-026119290303100608","unstructured":"PrakashA., MedcofJ.C. & TennantA.D. (1971). Paralytic shellfish poisoning in eastern Canada.\n                      Bulletin 177, Fisheries Research Board of Canada."},{"key":"bibr19-026119290303100608","first-page":"547","volume":"69","author":"Park D.L.","year":"1986","journal-title":"Journal of the Association of Official Analytical Chemists"},{"key":"bibr20-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1007\/BF02523835"},{"key":"bibr21-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1002\/nt.2620020511"},{"key":"bibr22-026119290303100608","doi-asserted-by":"publisher","DOI":"10.2331\/suisan.50.1413"},{"key":"bibr23-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1002\/etc.5620100305"},{"key":"bibr24-026119290303100608","first-page":"384","volume":"72","author":"McCulloch A.W.","year":"1989","journal-title":"Journal of the Association of Official Analytical Chemists"},{"key":"bibr25-026119290303100608","unstructured":"Anon. (2002). Updating Report on the Atypical DSP Results in Cockles. Website http:\/\/www.foodstandards.gov.uk\/multimedia\/pdfs\/note021202rep.pdf (Accessed 20.9.03)."},{"key":"bibr26-026119290303100608","unstructured":"FSA (2003). Website http:\/\/www.food.gov.uk\/multimedia\/pdfs\/shellaudittable.pdf (Accessed 3.10.03)."},{"key":"bibr27-026119290303100608","unstructured":"Anon. (2002). Cockles pose credibility test for FSA. Environmental Health News, May 3. Website http:\/\/www.ehn-online.com\/cgi-bin\/news\/editorial5\/EpkpuFFASzCSNkkD.html (Accessed 20.8.03)."},{"key":"bibr28-026119290303100608","unstructured":"MakinH.L.J. (2003). An Audit of Methods and Procedures for Lipophilic DSP Toxin Analysis Used by Laboratories at CEFAS, FRS and DARD, which Undertake the Statutory Monitoring of Shellfish Toxins in the UK, 110 pp. Website http:\/\/www.food.gov.uk\/multimedia\/pdfs\/shellmakinreport.pdf (Accessed 3.11.03)."},{"key":"bibr29-026119290303100608","unstructured":"FSA (2003). Investigations to Assess Whether Diethyl Ether or Acetone Carry-over During the DSP Standard Operating Procedure is Responsible for the Atypical Response in Mice. Website http:\/\/www.food.gov.uk\/multimedia\/pdfs\/shelletherpaper.pdf (Accessed 3.11.03)."},{"key":"bibr30-026119290303100608","first-page":"838","volume":"23","author":"Balls M.","year":"1995","journal-title":"ATLA"},{"key":"bibr31-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1258\/002367798780600016"},{"key":"bibr32-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1258\/002367799780487823"},{"key":"bibr33-026119290303100608","first-page":"383","volume-title":"Toxic Dinoflagellates","author":"Hamano Y.","year":"1985"},{"key":"bibr34-026119290303100608","first-page":"437","volume-title":"Mycotoxins and Phycotoxins","author":"Edebo L.","year":"1988"},{"key":"bibr35-026119290303100608","unstructured":"DennisonN., ZuurG., PetrieJ., TurrifffJ., KinnearS. & BourkeD. (2002). Pilot investigation of the use of general anesthesia to refine the statutory mouse bioassay for paralytic shellfish poison. In Program and Abstracts, Fourth World Congress on Alternatives and Animal Use in the Life Sciences, New Orleans, LA, USA, p. 119. Alternatives Congress Trust. Website http:\/\/www.worldcongress.net\/abstract-book\/contents.htm (Accessed 4.12.03)."},{"key":"bibr36-026119290303100608","volume-title":"Humane Endpoints in Animal Experiments for Biomedical Research.","author":"Hendriksen C.F.M.","year":"1999"},{"key":"bibr37-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1093\/ilar.43.Suppl_1.S112"},{"key":"bibr38-026119290303100608","first-page":"421","volume":"26","author":"Bottrill K.","year":"1998","journal-title":"ATLA"},{"key":"bibr39-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1016\/0278-6915(90)90117-6"},{"key":"bibr40-026119290303100608","first-page":"283","volume":"26","author":"Festing M.F.W.","year":"1998","journal-title":"ATLA"},{"key":"bibr41-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1093\/ilar.43.4.191"},{"key":"bibr42-026119290303100608","first-page":"203","volume-title":"Guidelines for Mutagenicity Testing, Part II, Supplementary Tests, Mutagens in Body Fluids and Excreta, Nitrosation Products","author":"Combes R.D.","year":"1984"},{"key":"bibr43-026119290303100608","unstructured":"Anon. (2002). Report of a Mission carried out in the United Kingdom from 8 to 17 July 2002 Regarding the Implementation of Council Directive 91\/492\/EEC (Live Bivalve Molluscs) and Council Directive 91\/493\/EEC (Fishery Products). DG (SANCO)\/8614\/2002 \u2014 MR final, 19 pp. Brussels, Belgium: European Commission. Website http:\/\/europa.eu.int\/comm\/food\/fs\/inspections\/vi\/reports\/united_kingdom\/vi_rep_unik_8614\u20132002_en.pdf (Accessed 28.11.03)."},{"key":"bibr44-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1016\/S0041-0101(01)00192-1"},{"key":"bibr45-026119290303100608","first-page":"71","volume-title":"The Design of Animal Experiments \u2014 Reducing the Use of Animals in Research Through Better Experimental Design","author":"Festing M.F.W.","year":"2002"},{"key":"bibr46-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1093\/ilar.43.Suppl_1.S69"},{"key":"bibr47-026119290303100608","first-page":"365","volume":"30","author":"Combes R.","year":"2002","journal-title":"ATLA"},{"key":"bibr48-026119290303100608","first-page":"587","volume":"7","author":"Sullivan J.","year":"1988","journal-title":"Journal of Shellfish Research"},{"key":"bibr49-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1016\/0041-0101(92)90517-9"},{"key":"bibr50-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1016\/0041-0101(93)90205-W"},{"key":"bibr51-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1016\/0041-0101(93)90359-Q"},{"key":"bibr52-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1002\/nt.2620020407"},{"key":"bibr53-026119290303100608","unstructured":"JellettJ.F., SilvaM.A., LaycockM.V., BellandE.R., BishopP.C., QuilliamM.A. & CembellaA.D. (2001). MIST Alert\u2122 for PSP and ASP: rapid tests for detection of marine toxins in phytoplankton monitoring.\n                      Abstract Presented at Second International Conference on Harmful Algae Management and Mitigation\n                      , Qingdao, China, 12\u201316 November. Website http:\/\/vm.cfsan.fda.gov\/~frf\/hamm01ta.html (Accessed 4.12.03)."},{"key":"bibr54-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1016\/S0041-0101(02)00153-8"},{"key":"bibr55-026119290303100608","author":"Laycock M.V.","year":"2000","journal-title":"Proceedings of IX International Conference on Harmful Algae"},{"key":"bibr56-026119290303100608","first-page":"455","volume":"21","author":"Mackintosh F.H.","year":"2002","journal-title":"Journal of Shellfish Research"},{"key":"bibr57-026119290303100608","first-page":"632","volume":"85","author":"Mackintosh F.H.","year":"2002","journal-title":"Journal of the Association of Official Analytical Chemists"},{"key":"bibr58-026119290303100608","unstructured":"Anon. (2003). PSP field test kit close to FDA approval. Website http:\/\/www.pcsga.org\/LongLines\/archive\/News_Archive.asp?IssueNum=112 (Accessed 3.11.03)."},{"key":"bibr59-026119290303100608","unstructured":"Anon. (2003). Result of ISSC consideration.\n                      ISSC Committee Report of 4 August, 2003.\n                      Lab Methods Review Committee."},{"key":"bibr60-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1002\/1522-7189(199911\/12)7:6<393::AID-NT82>3.0.CO;2-C"},{"key":"bibr61-026119290303100608","first-page":"745","volume":"27","author":"Combes R.","year":"1999","journal-title":"ATLA"},{"key":"bibr62-026119290303100608","first-page":"1152","volume":"280","author":"Laidley C.W.","year":"1997","journal-title":"Journal of Pharmacology and Experimental Therapeutics"},{"key":"bibr63-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1016\/S0041-0101(99)00163-4"},{"key":"bibr64-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1016\/S0003-2670(02)00597-4"},{"key":"bibr65-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1042\/bj2840539"},{"key":"bibr66-026119290303100608","first-page":"270","volume":"4","author":"Lee E.Y.","year":"1999","journal-title":"Frontiers in Bioscience"},{"key":"bibr67-026119290303100608","doi-asserted-by":"publisher","DOI":"10.2741\/A461"},{"key":"bibr68-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1006\/bbrc.2000.2459"},{"key":"bibr69-026119290303100608","unstructured":"Anon. (2003). Foodborne Pathogenic Microorganisms and Natural Toxins Handbook. Rockville, MD, USA: FDA. Website http:\/\/www.sfsan.fda.gov\/~mow\/chap37.html (Accessed 3.11.03)."},{"key":"bibr70-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1289\/ehp.5760"},{"key":"bibr71-026119290303100608","first-page":"225","volume":"31","author":"Combes R.D.","year":"2003","journal-title":"ATLA"},{"key":"bibr72-026119290303100608","first-page":"305","volume":"83","author":"Ledoux M.","year":"2000","journal-title":"Journal of the Association of Official Analytical Chemists"},{"key":"bibr73-026119290303100608","first-page":"352","volume":"23","author":"Combes R.D.","year":"1995","journal-title":"ATLA"},{"key":"bibr74-026119290303100608","first-page":"489","volume":"31","author":"Combes R.D.","year":"2003","journal-title":"ATLA"},{"key":"bibr75-026119290303100608","volume-title":"Naturally cccurring toxicants in food.","author":"Anon.","year":"1995"},{"key":"bibr76-026119290303100608","first-page":"67","volume-title":"OECD Environment, Health & Safety Publications Series on Testing and Assessment","author":"OECD","year":"2003"},{"key":"bibr77-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1080\/02652039309374131"},{"key":"bibr78-026119290303100608","unstructured":"BriggsL.R., MilesC.O., GarthwaiteI., RossK.M., TowersN.R., SamdalI., AasenJ., TorgersenT., AuneT. & PetersenD. (2002). Immunoassays for the analysis of yessotoxins.\n                      Abstract presented at 4th International Conference on Molluscan Shellfish Safety\n                      , June 4\u20138, Santiago, Spain. Website http:\/\/www.atlanticocongresos.com\/moluscos\/ (Accessed 4.12.03)."},{"key":"bibr79-026119290303100608","unstructured":"O'NeillS., GallacherS. & RiddochI. (2004). Suitability of a saxitoxin ELISA kit for use as a semiquantitative pre-screen for PSP toxins in\n                      Mytilus edulis\n                      and\n                      Pecten maximus\n                      , from UK waters, in press."},{"key":"bibr80-026119290303100608","unstructured":"Integrin (2003). Website http:\/\/www.integrin.co.uk\/seasafety.html (Accessed 7.10.03)."},{"key":"bibr81-026119290303100608","unstructured":"R-Biopharm (2003). Website http:\/\/www.r-biopharm.com (Accessed 7.10.03)."},{"key":"bibr82-026119290303100608","unstructured":"Bioresearch Ireland (2003). Website http:\/\/www.marine.ie\/mrm\/section1\/sec1_page12.html (Accessed 23.8.03)."},{"key":"bibr83-026119290303100608","first-page":"327","volume-title":"Mycotoxins and Phycotoxins","author":"Lee J.S.","year":"1988"},{"key":"bibr84-026119290303100608","first-page":"555","volume":"78","author":"Quilliam M.A.","year":"1995","journal-title":"Journal of the Association of Official Analytical Chemists"},{"key":"bibr85-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1016\/S0041-0101(01)00098-8"},{"key":"bibr86-026119290303100608","doi-asserted-by":"publisher","DOI":"10.1016\/S0041-0101(01)00097-6"},{"key":"bibr87-026119290303100608","unstructured":"CowlesJ., ConwayB. & MussellC. (2003). The simultaneous determination of ASP and DSP shellfish toxins using LC\/MS\/MS.\n                      Abstract Presented at FECS Euro Food Chem XII Symposium, Strategies for Safe Food\n                      , Brugge, Belgium, 24\u201326 September 2003. Website http:\/\/allserv.rug.ac.be\/EFCXII\/ (Accessed 4.12.03)."},{"key":"bibr88-026119290303100608","first-page":"49","volume-title":"Molluscan Shellfish Safety","author":"Aarsen J.","year":"2003"},{"key":"bibr89-026119290303100608","first-page":"1336","volume":"79","author":"Honkanen R.E.","year":"1996","journal-title":"Journal of the Association of Official Analytical Chemists"}],"container-title":["Alternatives to Laboratory Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.sagepub.com\/doi\/pdf\/10.1177\/026119290303100608","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.sagepub.com\/doi\/pdf\/10.1177\/026119290303100608","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,28]],"date-time":"2026-04-28T12:40:53Z","timestamp":1777380053000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.sagepub.com\/doi\/10.1177\/026119290303100608"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2003,11]]},"references-count":89,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2003,11]]}},"alternative-id":["10.1177\/026119290303100608"],"URL":"https:\/\/doi.org\/10.1177\/026119290303100608","relation":{},"ISSN":["0261-1929","2632-3559"],"issn-type":[{"value":"0261-1929","type":"print"},{"value":"2632-3559","type":"electronic"}],"subject":[],"published":{"date-parts":[[2003,11]]}}}