Contingency Plan for Hawaiian Monk Seal Unusual Mortality Events

A-3
1. If endoparasites are observed, collect immediately. Rinse, relax, fix, and store as described in
the parasitology section below. Be sure parasites are assigned the same specimen number as the
scat from which it is derived, but give each specimen a unique specimen sub-number.
2. Collect entire scat (or what's remaining if sub-sample(s) for parasitology are taken) in whirlpaks
or ziplocs. Be sure to include legible specimen labels both inside (using waterproof labels) and
outside the bags. Take care to ensure that the inside labels are visible from the outside.
3. Once any parasitology sub-samples have been collected, add 20% liquid detergent, 80% sea
water solution to scat sample to begin the degreasing process and to kill some of the pathogens.
Add just enough of this solution to keep samples moist. Remove as much air as possible from bags
before storing them in buckets. Be sure the samples are stored double bagged and are kept upright
in buckets to prevent leaks.
B. SPEWS
Spews (vomitus) are collected for prey species determination and parasite studies. They may also be used
to detect ciguatera (a naturally occurring biotoxin found in dinoflagellates) in tissues of certain prey
species. Spews are often found near seal haul out tracks and may contain partially digested eels, fish,
lobster carapaces, octopus, or whole prey items. Note: seabirds also spew squid, flying fish, and other prey
species, so be sure to closely observe seal tracks in the area the spew is found and collected.
1. If endoparasites are observed in the spew, collect them immediately and preserve as described in
the parasitology section below. Be sure parasites are assigned the same specimen number as the
spew from which it was derived, but give each specimen a unique specimen sub-number.
2. Next, collect a representative sub-sample of spew that is about 300-400 gm (100 gm is
approximately hamburger sized) for ciguatera analysis. Once the representative sample is
collected, split this sub-sample up into ~50 gm units and place in whirlpaks. Assign this spew subsample
the same specimen number as the spew from which it was derived, but give only ONE
unique specimen SUB-number to the representative sample, even though it is being split into
multiple bags. In addition to recording specimen number and sub-number on inner and outer
labels, note bag number and total number of bags for one sub-sample on each individual bag (1 of
5, 2 of 5, etc.) Be sure to also record the total number of bags for one spew sub-sample in the
"notes" field of the specimen collection summary form. NOTE: the splitting of the sample is
necessary to store the samples in liquid nitrogen. BE SURE the split samples will fit easily into
and out of the liquid nitrogen dewar. It can be very difficult, if not impossible, to remove large
items from the dewars due to their internal design.
3. Once any parasites sub-samples and the ciguatera sub-sample have been collected, store
remaining spew DRY in a whirlpak or ziplock, double bagged. Clearly label the outside of the
bag. Be sure to also include a legible specimen label inside (using waterproof labels) the bag.
Arrange the inside label so that it is visible from the outside. No preservation is necessary. Be
sure that the specimen is packed so that it is stored upright in buckets for storage and
shipment.
C. PARASITES
Polyvinyl Alcohol (PVA) fixed fecal specimens: Collect a minimum of 10 and a maximum of 30 PVA
fixed scat samples from fresh scats from known animals. Collection efforts should be focused on samples
from juvenile animals and collection of repeat samples from the same individuals.