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

Journal of Great Lakes Research

By:
, , , ,
DOI: 10.1016/j.jglr.2010.07.005

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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.

Additional Publication Details

Publication type:
Article
Publication Subtype:
Journal Article
Title:
Comparison of thiaminase activity in fish using the radiometric and 4-nitrothiophenol colorimetric methods
Series title:
Journal of Great Lakes Research
DOI:
10.1016/j.jglr.2010.07.005
Volume
36
Issue:
4
Year Published:
2010
Language:
English
Larger Work Type:
Article
Larger Work Subtype:
Journal Article
First page:
641
Last page:
645
Number of Pages:
5