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A Method for Measuring Total Thiaminase Activity in Fish Tissues
Author(s) -
Zajicek James L.,
Tillitt Donald E.,
Honeyfield Dale C.,
Brown Scott B.,
Fitzsimons John D.
Publication year - 2005
Publication title -
journal of aquatic animal health
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.507
H-Index - 52
eISSN - 1548-8667
pISSN - 0899-7659
DOI - 10.1577/h03-083.1
Subject(s) - biology , forage , forage fish , fish <actinopterygii> , incubation , substrate (aquarium) , invertebrate , zoology , ecology , fishery , biochemistry
An accurate, quantitative, and rapid method for the measurement of thiaminase activity in fish samples is required to provide sufficient information to characterize the role of dietary thiaminase in the onset of thiamine deficiency in Great Lakes salmonines. A radiometric method that uses 14 C‐thiamine was optimized for substrate and co‐substrate (nicotinic acid) concentrations, incubation time, and sample dilution. Total thiaminase activity was successfully determined in extracts of selected Great Lakes fishes and invertebrates. Samples included whole‐body and selected tissues of forage fishes. Positive control material prepared from frozen alewives Alosa pseudoharengus collected in Lake Michigan enhanced the development and application of the method. The method allowed improved discrimination of thiaminolytic activity among forage fish species and their tissues. The temperature dependence of the thiaminase activity observed in crude extracts of Lake Michigan alewives followed a Q 10 = 2 relationship for the 1–37°C temperature range, which is consistent with the bacterial‐derived thiaminase I protein.