Comparison of thiaminase activity in fish using the radiometric and 4-nitrothiophenol colorimetric methods

Honeyfield, D. C.; Hanes, J. W.; Brown, L.; Kraft, C. E.; Begley, T. P.

Abstract:

Thiaminase induced thiamine deficiency occurs in fish, humans, livestock and wild animals. A non-radioactive thiaminase assay was described in 2007, but a direct comparison with the radioactive 14C-thiamine method which has been in use for more than 30years has not been reported. The objective was to measure thiaminase activity in forage fish (alewife Alosa pseudoharengus, rainbow smelt Osmerus mordax, and slimy sculpin Cottus cognatus) consumed by predators that manifest thiamine deficiency using both methods. Modifications were made to the colorimetric assay to improve repeatability. Modification included a change in assay pH, enhanced sample clean-up, constant assay temperature (37??C), increase in the concentration of 4-nitrothiophenol (4NTP) and use of a spectrophotometer fitted with a 0.2cm cell. A strong relationship between the two assays was found for 51 alewife (R2=0.85), 36 smelt (R2=0.87) and 20 sculpin (R2=0.82). Thiaminase activity in the colorimetric assay was about 1000 times higher than activity measured by the radioactive method. Application of the assay to fish species from which no thiaminase activity has previously been reported resulted in no 4NTP thiaminase activity being found in bloater Coregonus hoyi, lake trout Salvelinus namaycusch, steelhead trout Oncorhynchus mykiss or Chinook salmon Oncorhynchus tshawytscha. In species previously reported to contain thiaminase, 4NTP thiaminase activity was measured in bacteria Paenibacillus thiaminolyticus, gizzard shad Dorosoma cepedianum, bracken fern Pteridium aquilinum, quagga mussel Dreissena bugensis and zebra mussels D. polymorpha. ?? 2010.

Genre:

Article

ProdID:

70037668

Citation Author:

Honeyfield, D. C.; Hanes, J. W.; Brown, L.; Kraft, C. E.; Begley, T. P.

Citation Contributing Office:

Citation Datum:

Citation Day:

Citation Edition:

Citation Editor:

Citation End Page:

645

Citation Issue:

Citation Keywords:

Citation Language:

English

Citation Larger Work Title:

Journal of Great Lakes Research

Citation LatN:

Citation LatS:

Citation LonE:

Citation LonW:

Citation Month:

Citation No Pagination:

Citation Number Of Pages:

Citation Online Only Flag:

Citation Phsyical Description:

Citation Projection:

Citation Public Comments:

Citation Publisher:

Citation Series:

Citation Series Code:

Citation Series Number:

Citation Search Results Text:

Comparison of thiaminase activity in fish using the radiometric and 4-nitrothiophenol colorimetric methods; 2010; Article; Journal; Journal of Great Lakes Research; Honeyfield, D. C.; Hanes, J. W.; Brown, L.; Kraft, C. E.; Begley, T. P.