AN IMMUNOLOGICAL STUDY OF PREDATION ON HATCHERY-REARED, JUVENILE RED DRUM (SCIAENOPS-OCELLATUS) - PREPARATION AND ASSAYS OF A RED DRUM-SPECIFIC PROTEIN FOR PREDATOR-PREY EXPERIMENTS
Dr. Schultz et Me. Clarke, AN IMMUNOLOGICAL STUDY OF PREDATION ON HATCHERY-REARED, JUVENILE RED DRUM (SCIAENOPS-OCELLATUS) - PREPARATION AND ASSAYS OF A RED DRUM-SPECIFIC PROTEIN FOR PREDATOR-PREY EXPERIMENTS, Journal of experimental marine biology and ecology, 189(1-2), 1995, pp. 233-249
This immunological study is part of a program to test the feasibility
of artificial stock enhancement using hatchery-reared red drum (Sciaen
ops ocellatus) (L.) and specifically to determine the predation mortal
ity of newly released fish. Initially, a polyvalent antiserum was prod
uced in a goat by multiple injections of a soluble red drum extract. I
n comparative experiments with soluble extracts of 12 different fishes
and three different invertebrates found in the same environment, usin
g sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the go
at polyvalent antiserum for Western blots, an 80 kDa ''red drum specif
ic'' protein was selected that was absent from the other species, The
protein was present as two isoforms in fingerlings, but as one protein
in older fish, The protein was purified by ammonium sulfate fractiona
tion, and column chromatography with the Fast Protein Liquid Chromatog
raphy system. A second goat was immunized with the highly purified red
drum 80 kDa protein to obtain a specific antiserum for immunoassays.
Cross-reactivity in Western blots with pre-immune and immune goat sera
and unrelated fish proteins was removed from the sera by solid phase
adsorption. The adsorbed goat anti-80 kDa glycoprotein serum was used
in predator-prey studies to identify partially digested-visually unrec
ognizable red drum. In controlled laboratory feeding experiments, the
80 kDa protein was detected in Western blots of stomach contents after
1 and 2 h of digestion, but not after 4 h. Western blots after 2 h sh
owed immunoreactive breakdown-products from the 80 kDa protein in stom
ach contents. The methods with the red drum protein appear applicable
to any fish for specific identification in predator gut contents by im
munological procedures.