Measurement of arginine kinase activity in hemolymph ...



Title Measurement of arginine kinase activity in hemolymph of American lobsters
Author(s) Barbara S. Horney, A. L. MacKenzie, Richard J. Cawthorn, R. J. MacMillan, C. Morris
Journal Journal of Aquatic Animal Health
Date 2001
Volume 13
Issue 4
Start page 328
End page 333
Abstract The concentration of arginine kinase (AK) in the hemolymph of lobsters may aid in the identification of muscle diseases in these animals in the same way that creatine kinase activity in blood plasma does in mammalian species. Both manual and automated analysis methods were developed for measuring AK levels in the hemolymph of the American lobster Homarus americanus. A commercial reagent that is used to measure creatine kinase in human blood serum was modified by substituting phospho-L-arginine for creatine phosphate as the substrate to allow measurement of the AK enzyme. Stability of the enzyme in lobster hemolymph plasma was less than 3 h at room temperature (22 degree C), 6 h when refrigerated (2-5 degree C), and 24 h when frozen (-20 degree C). Comparisons were made between the manual and automated protocols using three lobster hemolymph plasma samples. The average recovery in the latter relative to the former was 103%. The automated assay was linear up to 1,940 U/L. Precision studies were conducted using the automated procedure with low and high AK plasma samples. Coefficients of variation were less than 2% for the within-run (N = 15) and less than 3.5% for the between-run assays (N = 7). A nonparametric description (range, 9-3,322 U/L; median, 111 U/L; 25th-75th percentiles, 71-304 U/L) was applied to the data from the automated method for AK concentrations in lobster plasma using 95 healthy lobsters obtained during the 1995 spring season in Lobster Fishing Area 26A, Prince Edward Island. No significant differences in AK levels were detected between male and female lobsters. Identifiable increases in hemolymph AK activity were detected in 5 out of 10 lobsters 44 d after they were experimentally infected with the ciliate parasite Anophryoides haemophila. Our applications will facilitate consistent analysis of AK levels in the hemolymph of the American lobster.
DOI 10.1577/1548-8667(2001)013<0328:MOAKAI>2.0.CO;2
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