Vol. 53 - Alaska Resources Library and Information Services

Battelle's Pacific Northwest Division in Richland, Washington.
After multiplication of the soft tissue dry weights by the caloric
values, summing the caloric intakes for all items gave the daily ration in
calories per g crab wet weight per day. The percentage of this total
caloric intake that each item contributed was then calculated.
IMMUNOASSAY
Because the grinding of the crab's food by the gastric mill (Warner,
1977) renders prey unidentifiable, visual examination of gut contents
suffers bias. To identify prey entirely missed by conventional analysis,
immunological examination of gut contents modified from Feller et al
(1979) was performed. Briefly, probable macro and meiofaunal prey items
were collected as described above. On shipboard these organisms were
isolated in seawater filtered to 45 microns. After up to 7 days to clear
their stomachs of foreign protein the isolated organisms were flash
frozen. In the laboratory, whole organism extracts were prepared by
grinding frozen prey in a chilled buffer solution. Protein concentrations
in the extracts were determined spectrophotometrically against albumin
standards. Following standard protocols (Kenny, 1971) extracts of known
protein concentration were injected into rabbits to produce antisera of
varying specificity.
To determine self and cross reactions and reactions between the
antisera and gut contents, standard Ouchterlony (double immuno diffusion
precipitin) tests were performed. In this test the antigen and the
antisera were allowed to diffuse towards each other through a supporting
medium of 0.5 percent agarose gel on glass microscope slides for 48 hours.
When a soluble antigen (prey organism extract or stomach contents) reacted
with its specific antibody (antisera to prey organisms), a precipitate was
formed where they met in optimal proportions. The opaque precipitin lines
were then stained with Coomassie Brilliant Blue R and counted. Based on
the number of precipitin reaction lines encountered, a matrix table of
self and cross reactions was developed similar to Feller's.
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