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A Non-Lethal Procedure for the Determination of Organic Contaminant Concentrations
in Lake Sturgeon (Acipenser fulvescens)
Introduction:
Lake sturgeon (Acipenser fulvescens) is an ancient species that thrived in the Laurentian Great
Lakes for thousands of years before humans colonized the region but once we began fishing
these waters, their habit of destroying fishing nets gave sturgeon nuisance species status among
fishermen and many of them began to kill sturgeon whenever and wherever possible.
Eventually, uses were found for various tissues especially the meat, roe, swim bladder, and hide.
Both factors resulted in tremendous fishing pressure that, along with a long reproductive cycle,
combined to produce a decline in the population to dangerously low levels. The species is now
listed as threatened and is protected in the Michigan waters of the Great Lakes under the
Michigan Endangered Species Act of 1994 and is listed as Special Concern, Threatened, or
Endangered in all of the eight Great Lakes states. Harvest is banned or severely restricted
throughout its range in the basin.
Protection under the Michigan Endangered Species Act has made it impossible to incorporate
sturgeon into routine contaminant monitoring programs that typically require the sacrifice of the
fish but which produce data critical to a better understanding of a species’ overall health.
Because of this protection, there is very little data published enumerating the contaminant levels
in sturgeon.
From several perspectives, knowledge of contaminant levels in lake sturgeon is vital. From an
ecosystem perspective, this long-lived species will act as a sink for many contaminants since it’s
armor and size afford it protection from predation. Tracing the movement of target chemicals
into sturgeon is therefore important. Most species of sturgeon are today prized for their meat and
roe. Lake sturgeon are no exception and there is a considerable demand for these food products.
Although the species is afforded protection in the United States waters of the Great Lakes,
Canada allows a quota each year to meet the public demand. It is therefore necessary to obtain
baseline data and establish monitoring programs to assess the risk of human. Lastly,
contaminant data is needed to assess the general health of the species, to help predict its potential
for recovery, and will be used in the management decision-making process for lake sturgeon
populations.
To this end, a non-lethal method has been developed to sample lake sturgeon tissue. This
method, once deployed, will provide invaluable contaminant data without harming the species’
population or violating the Endangered Species Act.
The procedure involves the use of a 0.5” (12.7 mm) i.d. steel tube, sharpened at one end, to take
a core sample through the fish. The core sample is taken at the midpoint of the dorsal fin on the
left side. A small incision makes it easier to start the sampling tube into the dorsal surface.
After this, the tube is driven straight down through the tail and ventral skin. No primary closure
is required. Wounds should heal without serious complication within approximately eight
weeks.
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The method is suitable for most organochlorine contaminant analysis and has applicability to
other species of sturgeon. Because the method employs a standardized location for sampling, it
may not be applicable to non-acipenserid species. Its use should yield 4-6 grams of tissue,
sufficient for most modern instrumentation to quantify contaminant levels.
Contaminant data presented here were derived from fish provided by the US Fish and Wildlife
Service and from field tests conducted in June of 2002. Specific tissues included muscle, gonad,
liver, and blood. The analytes included 80 PCB congeners including the most toxic planar
congeners, 21 pesticides, and Toxaphene.
Methods:
Several sturgeons were dissected in the lab to obtain tissue samples for comparative contaminant
analysis, to determine pertinent anatomy, and to test various methods and locations of sampling.
For this study, muscle, blood, liver, and gonads were considered for sampling purposes. We
looked for the best location to collect each of these tissues and considered the ramifications of
each. After determining the appropriate tissue, experiments were run to determine the tool and
method that would yield the most tissue while posing the least risk of mortality and sample
cross-contamination. Efforts quickly focused on a tunneling device of standard dimension to
ensure comparable samples. Samples of ten grams, standard for our GC/MS contaminants
protocols, were compared with one-gram samples to find out if lower mass samples would pose
detection limit problems.
After determining the tissue, location, and method of sampling, experiments on the effects of
direct application of a concentrated anesthesia were conducted. A 10% solution of clove oil in
ethanol was diluted to concentrations of 12,800 mg/L, 1,600 mg/L, and 800 mg/L. A spray
bottle set to a very fine mist was used to apply the three concentrations to the gills of 4 4-yearold lake trout. Efficacy of each concentration was determined based on the fish’s reaction to
pain, in this case a dissecting needle inserted into the flesh.
Field tests occurred on 11 June 2002 to determine if the sampling method was indeed non-lethal.
We used 4 sturgeon as experimental fish and two more for control fish. All fish for this study
were captured on 7 June 2002 or 10 June 2002 in Lake Huron near the headwaters of the St.
Clair River at Point Edward, Ontario. The holding facility was a cement enclosure measuring
approximately 2.4m x 2.4m x 1.2m deep. Water, drawn directly from the St. Clair River, was
circulated through the tank at approximately 50 liters/minute.
The procedure during the field test was as follows: One control fish was removed from the
holding tank, measured, weighed, tagged for identification (external Floy tag), and returned to
the holding area. The second control fish was treated in the same manner with the addition of
clove oil anesthesia at a concentration of 800 mg/L. These control fish provided a distinction
between any mortality caused by the sampling method or gill damage caused by anesthesia from
mortality due to water quality issues or communicable disease in the holding area. Next, I tested
the method, including anesthesia, on four experimental fish. Here again, the fish were tagged
externally, and lengths and weights were recorded prior to testing. The full details of the method
are given in Appendix A and will not be repeated here.
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For comparison, one extra fish was sampled using a method developed for smaller species by the
Ontario Ministry of Natural Resources (ONMNR). The ONMNR method was accomplished by
making a 1 cm incision in the thickest portion of the muscle, parallel to the midline. A second
incision was started at the posterior end of the first and extended ventrally approximately 1 cm.
Both incisions were also 1 cm deep. After the area was undercut, the tissue attached to the skin
flap was grasped and peeled away from the skin, which was then pressed back down into the
wound.
Also during the field tests, several fish were sacrificed and tissue sample were collected
including muscle, liver, and gonad. All of the samples taken on 11 June 2002 were put on ice
and returned to the lab where they were frozen until analysis. The samples in this study were
analyzed via GC/MS using Selected Ion Monitoring in Negative Chemical Ionization mode.
Specifically, GLSC uses an HP Model 5973 GC/MS. The column used was a capillary column
JW DB-5, 30m x 0.25mm i.d. x 0.25 m thickness. The following analytes were quantitated:
80 PCB congeners, Total PCB, Pentachlorobenzene, alpha-HCH (Hexachlorcyclohexane),
Hexachlorobenzene, PCPME (Pentachlorophenylmethylether), Lindane, Aldrin, Dachtal,
Octachlorostyrene, b-Heptachlor Epoxide, Oxychlordane, trans-Chlordane, cis-Chlordane, transNonachlor, cis-Nonachlor, Dieldrin, Endrin, pp’-DDD, pp’-DDE, pp’-DDT, Photo-Mirex,
Mirex, the hexa through deca-chlorinated Toxaphene homologues, and Total Toxaphene.
The test fish were examined on 18 June 2002 and, again on 27 June 2002 at which time they
were released into the headwaters of the St. Clair River.
Results:
Table 1 enumerates the analytical data from the chemical analyses. Total PCB is calculated as
the sum of the 80 congeners. Toxaphene is presented by chlorination level, Total Toxaphene
being the sum of the five levels. The individual peaks that belong to each chlorination level are
of no significance and have been omitted. All Table 1 results are given in units of ng/g, wet
weight. Values of ‘0’ can be taken to mean exactly 0.00. For values over 99.99, only the three
digits to the left of the decimal are significant. I have provided no data for the fish sampled
using the ONMNR method because it was done solely for wound comparison purposes.
With regard to the fish designations in Table 1: Lab Fish 1-3 were dead fish from which samples
were collected in the lab for comparative analyses. For Lab Fish 1 and 2, collected at Manistee
in Lake Michigan in 1999, each tissue (blood and liver) was run twice to demonstrate consistent
results on duplicate runs. For Lab Fish 3, collected in Lake St. Clair in 2001, a 10-gram sample
of muscle was compared to a 1-gram sample to demonstrate that the lower amount was sufficient
for accurate analysis. Samples were also taken from two different areas of the testes and liver of
Lab Fish 3 to determine homogeneity of the contaminant loads in those tissues. Samples were
collected from the tip and mid-portions of those organs. All three of these fish exceeded our
instrumentation’s capacity to accurately weigh them. Fork length and commercial length
measurements were not collected for lab fish 3.
Test Fish 1-4 were the four experimental fish sampled using the final method on 11 June 2002.
Sacrificed Fish 1-8 were the extra fish, sacrificed for commercial purposes on 11 June 2002,
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from which samples of various tissues were also taken. Muscle samples from the four test
subjects were collected using the standardized location called for in the method. It was not
possible to collect samples from a consistent anatomic location from the eight sacrificed fish.
There is no discrete sex chromosome in fish therefore, sex determinations could not be made for
the four test fish or the two control fish. Efforts are underway to determine the gender of those
samples via genetic marker and results will be forwarded upon completion.
A total of 15 fish were sampled for various tissues during this study. In some cases, an emerging
issue contaminant, Polybrominated Diphenyl Ethers (PBDEs) is mentioned as points of interest
although analyses of this chemical group were limited and data are not provided.
Table 2 consists of the physical data corresponding to each fish in Table 1. This table also
contains physical data for the two control fish used on 11 June 2002 although these fish were not
sampled for chemical analyses.
Results from the tests on the lakes Michigan and St. Clair lab fish and the Lake Huron test fish
yielded the following results:
Lake Michigan 1999: Chemical analyses demonstrated that significant amounts of chlorinated
hydrocarbons were accumulated in the blood and liver of the sturgeon collected near Manistee in
1999. In spite of the fact that the age, gender, and length of collected fishes were different, their
PCB concentrations in the liver were similar, differing no more than 10% from 850 ng/g to 950
ng/g. The margin in blood samples was somewhat greater, varying by 20% from 800 ng/g to 980
ng/g. Nevertheless, these concentrations are much lower than lake trout sampled at
approximately the same time and location where whole fish and liver had values of about 1,800
ng/g and 7,500 ng/g respectively.
It is important to mention that in contrast with PCB accumulation by other fish species in Lake
Michigan, the distribution of congeners in lake sturgeon is relatively more even. The highest
abundance was found in congeners #153 and #163, corresponding to lake trout from Lake
Michigan, but the next highest congeners were #118, #84, #66, and #71 which do not contribute
significantly to lake trout or smelt contaminant loads. There are insignificant amounts of the
toxic planar congeners. Congener #126 did not exceed 0.5 ng/g in sturgeon whereas it tends to
be 10 ng/g in lake trout from the same location.
In the pesticide category, there is great variability in the liver concentrations of DDE,
Toxaphene, Mirex, and Total Chlordanes, depending on age and gender however, the
concentrations did not vary greatly in the blood samples. The octa and nona-chlorinated
Toxaphene homologues showed a greater relative abundance in lake trout as compared to lake
trout, which typically have more of the hepta-chlorinated homologues.
Emerging issue contaminants, especially Polybrominated Diphenyl Ether (PBDE) #47 and #99
showed very small concentrations in Manistee sturgeon at 1 ng/g and 0.2 ng/g respectively as
compared to lake trout. In contrast to lake trout, where concentrations of the various categories
is very consistent, PBDE concentrations in sturgeon exceed the concentrations of some of the
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more global contaminants, specifically Chlordanes, HCHs, Hexachlorobenzene, and most
industrial organochlorines.
Lake St. Clair 2001: A different picture of contaminants was observed in 2001 Lake St. Clair
samples. PCB concentrations were much lower in these fish as compared to Lake Michigan fish.
The concentrations for liver, testes, and muscle were 400-600 ng/g, ~650 ng/g, and ~425 ng/g
respectively. Even the very fatty muscle tissue had only 3,700 ng/g. Unlike Lake Michigan
sturgeon, these fish looked very much like most other biota from that area with respect to the
congener patterns. Congeners #153, #163, #180, and #187 were the most abundant. We found
only traces of Toxaphene in this fish and moderate concentrations of most other pesticides
including DDE, trans-Chlordane, and Oxychlordane.
Lake Huron 2002: A relationship was seen in the 2002 Lake Huron fish. The test subjects were
smaller on average than the eight sacrificed fish. As expected, the contaminant loads were lower
in muscle tissue, especially with regard to total PCB. An exponential relationship is noted (R 2 =
0.597) between total length and Total PCB within this group of twelve fish (Fig. 6). Because the
four test subjects could not be sampled for gonad and liver tissue, resulting in a more narrow
range of lengths, these relationships were much weaker. As expected, when the four test fish are
removed from the muscle analysis, the relationship becomes linear and shows practically no
goodness-of-fit (R2 = 0.0276). The three lab fish were similar in size to the sacrificed fish, were
from a different site, and had been frozen prior to sampling, and therefore lend no further insight.
Discussion:
It was quickly determined that the only possibilities for tissue sampling were muscle, blood,
gonad, or liver. The latter three tissues all have at least one problem in common: There is no
good way to indicate that a given fish has been previously sampled upon subsequent capture.
Repeated sampling of the same fish would not yield viable information on the health of the
species and may in fact endanger the health of the individual. Even external tagging would not
alleviate this problem in instances where a collector captures a fish bearing another collector’s
tag. Contacting the previous collector and determining if the tag corresponds to a sampled fish is
impractical at best. I did examine each of the three tissues to determine if there were extenuating
circumstances that would make these tissues worthwhile. There were not. The external scar left
by the new method is a clear indication of previous sampling.
Here is a brief synopsis of each of the three tissues I didn’t use, and why, followed by supporting
arguments for using muscle.
Liver: Although the liver is very easy to locate, this method would be too traumatic.. Its largest
lobe is invariably found at the midline between the distal ends of the medial ray of the pectoral
fins, when the fins are in the normal position. The one variable I can’t account for in sampling
the liver is blood loss. In many fish species, the liver has very elongated lobes and I’m sure the
distal ends of each have a relatively low blood supply. Sturgeon have a more compact liver
increasing the chance that I will sever a major blood vessel and cause substantial bleeding. It
may not be possible to stem that kind of bleeding. In a related incident, Dr. James Boase of the
US Fish and Wildlife Service has indicated that he ‘nicked’ a liver in his efforts to implant a
sonic transmitter in a lake sturgeon. That transmitter stopped moving soon after the fish’s
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release, indicating either the death of the fish or the expulsion of the tag, the latter being less
likely. Peritonitis is of concern anytime the body cavity is violated since sufficiently antiseptic
conditions cannot be achieved in the field. Figure 1 demonstrates the location, size and shape of
the liver.
As you can see from the data presented in Table 1, the liver contaminant content is wildly
variable. That would leave us without reliable whole-body estimates or trend data. It is also
apparent from the data on Lab Fish 3 (Table 1) that the location of the sample will greatly affect
the contaminant results.
Blood: While this tissue is also easily obtained using standard phlebotomy equipment, blood
content is variable depending on many factors and it wouldn’t be reliable. Again, we would not
be able to relate this to a whole-body burden or develop trend data.
Gonad: This tissue is too hard to find and too dangerous to take. If the individual is not close to
spawning, the gonads will likely be very small and hard to find. There won’t be an anatomical
landmark available to locate the incision since the state of the gonads isn’t known beforehand.
Sturgeon are believed to spawn only once every 4 to 6 years making it impossible to gauge the
reproductive state of the population even if sampling is undertaken at the same time each year.
In any specimen other than a ripe female, the researcher would have to resort to taking
reproductive tissue instead of the gametes themselves and although this may not kill the fish, it
may irreparably hinder the fish’s fecundity and raise the issue demonstrated in Table 1 where the
data are variable depending on the location of the sample. Further, the reproductive tissues are
fundamentally different between male and female giving rise to questions of comparability.
Peritonitis is a very real danger here as it is with liver sampling.
Muscle: Table 1 enumerates data supporting the decision to use muscle samples. The data
indicate that most modern analytical instruments using the same methods used by GLSC will be
able to achieve valid results from as little as one gram of tissue. Most other tissue types yield
results far higher than the muscle tissue and many of the results are inconsistent from fish to fish
whereas the muscle tissues are not inconsistent when the fish are of similar size. Choosing
tissues with more constant concentrations will allow for easier monitoring of the species.
Because of the problems with the other tissues, and since the muscle is of commercial value,
collection of muscle tissue for the purposes of this study is indicated.
The standardized location (next to the dorsal fin on the left side) helps ensure comparable results
between fish while avoiding damage to vital structures. The size of the individual fish will
determine exactly how much tissue is harvested, but I believe that the range should be 4-6 grams.
It was not possible to weigh the samples in the field because of logistics and sample
contamination issues. The samples had been frozen for a couple weeks before weights could be
taken and they were visibly dehydrated. This accounts for the weights given in Table 1, which
ranged from 2 to 3 grams in the four experimental fish.
Several configurations and alloys were considered with regard to the sampling tube. Brass was
considered first because it is easy to sharpen and could be obtained in a thinner gauge. Brass
however, contains large amount of lead that poses a health risk to the fish. This metal will also
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contaminate the samples and lend uncertainty to the analyses. The use of steel will eliminate
these issues and also make it possible to test for the presence of heavy metals, should the
researcher so choose.
A tube was tested with a toothed edge, an edge beveled such that it looked like a large syringe,
and with a straight edge. Anything other than a straight edged tube caused extensive muscle
damage and reduced the amount of tissue harvested. A sampling tube 12” (35 cm) long should
be adequate for most specimens. The tube should typically have to pass through 7.5 cm of tissue
on larger fish such as in Figure 4.
It appears that the myomere structure affects how well tissue can be sampled using this method
in that a direction other than dorsal-to-ventral yields far less tissue. Figure 2 shows a posteriorto-anterior approach. This particular picture shows an attempt that was shorter than that of the
finalized method shown in figure 3 however, all attempts of this method (many of them almost
as long as the tube) yielded less than 5 grams of tissue. Many yielded less than 2 grams.
Sampling anterior to posterior necessitates keeping the sampling tube deep in the tissue to
prevent cutting off the blood supply to the skin, thereby causing necrosis, and it was difficult to
keep the tube from violating the peritoneal cavity on longer attempts. Again, organ damage and
infection are a concern. This approach would be far more difficult to recover from and would
cause too much damage because of the number of myomeres through which the tube would pass.
Comparison of ten-gram to one-gram samples showed no significant differences, which supports
the decision to use the less dangerous, though slightly less fruitful, direction.
If the sampling tube is very sharp, penetration of the skin is accomplished with relative ease by
applying pressure and twisting quickly; however, on occasion, the tube will slip from its starting
position without fully penetrating the skin. It was found to be much easier to start with an
incision of about a little more than one centimeter. The tube can be partially inserted into the
opening and then driven down through the muscle and ventral skin. Figure 3, a photo taken 16
days after sampling, demonstrates this problem. Test fish number one was not started with an
incision and the sampling tube slipped on the first attempt whereas the entry point on test fish
three was begun with an incision and is very clean. The dorsal wounds on experimental fishes 24 all looked similar to the wound depicted in figure 3. The ventral exit point on all test fish was
circular in shape with a very clean edge, as depicted in figure 3.
No primary closure of the wound is needed. In fact, leaving the wound open and allowing it to
heal secondarily, facilitates proper drainage and reduces the chances of infection. Several
methods of closure were considered including sutures, medical grade staples, industrial grade
staples, and veterinary glue. Each was found to be either too expensive, time consuming, or
difficult. All were found to be unnecessary for this procedure.
Attention should be paid to water quality before using this method. Elevated levels of bacteria
may preclude the use of the method due to the increased chance of infection. In general, if there
are no restrictions on swimming or fishing in affect because of bacteria levels, there should be no
risk to the fish.
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No remarkable differences were noted between the condition of the fish on 27 June 2002 and the
pre-test condition in terms of their behavior and overall health. Minor cellulitis and redness was
noted in all cases at the time of release; however, none of the fish appeared to have a serious
infection and the wounds appeared to be healing. Blood loss during the procedure was
approximately 10cc and was not considered significant. Hemorrhaging ceased within minutes of
returning the fish to the holding tank after the procedure and I noted none thereafter.
The sampling method and standardized location leaves a scar that should clearly indicate
previous sampling. Although the dorsal scar may resemble a lamprey wound, especially if an
incision is not made to start the sample, it is highly unlikely that the fish would also have a
lamprey wound on the ventrum located precisely below the dorsal scar. I believe that most
researchers will also employ an external tag and will collect a fin ray for age analysis when they
collect samples via the new method. This will serve to further identify previously sampled fish.
See figure 4 (included in Appendix A as fig 1) for location and size of sampling site.
The method is quick and less invasive than the gonad and liver techniques. It had very little
effect on the fish’s ability to swim in the holding tanks and should heal within several weeks. I
have noted fish captured in the wild that have very large scars from encounters with various
sharp objects, which do not appear to have had any long-term affects and this method is much
less traumatic than any of those wounds. The presence of lamprey wounds, both fresh and
previously healed, that don’t appear to harbor significant infection is further evidence that this is
the most appropriate method.
The results of the ONMNR sampling method were far less encouraging. Examination of the fish
on 27 June showed considerable inflammation of the surrounding tissue and a skin flap that had
shriveled, was partially necrotic, and was unlikely to heal over the exposed muscle (Fig. 5). I
don’t believe that the method would have yielded the consistent mass of tissue achieved by the
final methodology. In lab tests, I took a one gram sample using a pyramid-shaped incision
having dimensions of 18mm x 18mm x 15mm and 17mm deep. The Ministry’s method would
yield no more tissue. Both of these similar methods had to be done on the more anterior portion
of the fish where the muscle was thicker and subsequently, there are no landmarks to standardize
the method.
Sturgeon are placid by nature, allowing extensive handling and surgical procedures without
becoming highly agitated. With this in mind, it was considered doubtful that anesthesia would
be a necessity. Initially it was considered solely for humane reasons. During the tests on 11
June 2002 however, it was found that sedation serves a vital purpose. Most other procedures I
have witnessed were conducted on the abdomen of the fish. When they do become agitated,
most of the thrashing is at the tail. This new sampling procedure involves the more posterior
portion of the fish and can become difficult and even dangerous if the fish thrashes with the
sampling tube still in its tail. The first test fish was not fully anesthetized and began to struggle
when pressure was applied to the sampling tube. This caused the tube to penetrate at a slight
angle, reducing the amount of tissue sampled and causing increased bleeding and skin damage.
That experience indicates that there is indeed a genuine need for sedation of some kind.
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The method of sedation is an adaptation of the anesthesia procedure commonly used at GLSC
wherein a solution of 10% clove oil in Ethanol is prepared. This stock solution is added to a
water bath at varying concentrations depending on the species, size and number of fish. With the
proper concentration, anesthesia is achieved within one minute and lasts up to several minutes.
Due to the size of the fish, it is not practical to make a water bath for the purpose of anesthetizing
a sturgeon. The procedure also needs to be simple and inexpensive such that it would allow for
ease of use on a boat where much of the sampling will undoubtedly be done. This made
fabricating a pumping system impractical as well. Instead, a direct application was developed,
via a spray bottle, using a much more concentrated solution to compensate for the smaller
volume and shorter exposure time.
Clove oil is not yet legally approved as a fish anesthetic but its use is widespread. MS-222
(Tricaine Methanesulfonate, a.k.a. Triple-2) is the more commonly used anesthesia. Clove oil is
safer for human handling, in my experience has a shorter recovery period, and is therefore
preferred.
Three concentrations were tested in the lab on 4-year-old lake trout. The first concentration of
12,800 mg/L (80 ml of a 10% stock solution in a 650ml bottle) was applied by spraying the gills
twice. The fish hemorrhaged and died almost instantly. Fish two was treated with one spray per
gill of a 1,600 mg/L solution (10ml stock solution). After less than two minutes, hemorrhaging
was noted and the corneas had considerable damage from the alcohol content, which was over
1.2%. The fish died moments later. Fishes three and four were given an 800 mg/L (0.658%
alcohol) dose. Fish three was sprayed twice on each gill while fish four was sprayed only once
but was also sprayed once on the left eye to test for corneal damage. Both were well sedated at
this concentration, both recovered quickly and although fish four showed evidence of bleeding in
the recovery tank, I believe this was from rough handling and not dehydration of the gills. There
was no evident damage to the eye of fish four. Although the final concentration did not produce
gross tissue damage in the test subjects, the method advises that the eyes be covered with a damp
rag and irrigated copiously during application. The residue must also be rinsed away from the
gills as soon as the procedure has finished. In the field, it took up to 8 sprays per gill on sturgeon
to achieve adequate sedation but this did not seem to pose a problem for the fish. Sedatives
should be delivered in two-to-four spray increments so that the minimum is used.
The test solutions were made using ethanol from a large supply (55 gallon drum) that the lab
uses for sample preservation and other purposes. I later discovered that the 10% clove oil
solution was contaminated with polybrominated diphenyl ethers (PBDEs), commonly used
organic flame-retardants that are one of the hot topics in the world of contaminant monitoring
and potentially a target compound for studies for which this sampling method was designed.
This was a qualitative study only and the specific concentrations are not germane. Analyzing the
solution was actually only an afterthought and no undiluted clove oil from the original bottle was
left to test for contaminants. Because the clove oil was reagent grade from Sigma, it is doubtful
that the clove oil itself was the source of the PBDEs. Ergo, using high quality alcohol can
eliminate the contamination. Two other points are notable: 1. This solution is meant only as a
minor sedation and it’s introduction, via the gills, should not be able to contaminate the tissue
sample so long as the collector takes care not to spray the solution on the sample, or the area to
be sampled, and washes their hands if they are exposed to the solution prior to sampling. 2.
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After the 10% solution is diluted in stream water to the 800mg/L working concentration, PBDE
levels in the resultant solution should not be measurably higher than the stream water alone.
Other precautions that would eliminate the problem include wearing rubber gloves during clove
oil administration and strictly separating the duties of anesthetist and sampler. It should be noted
that absolute alcohol, although usually of the highest quality, is a poor choice for this application.
It may contain carcinogens and may also, due to its affinity for moisture, pull contaminants from
the air into the container after opening.
Once the sampling procedure was completed and the sturgeon’s gills were flushed with water to
remove the excess anesthesia, the fish recovered within 30 seconds. As soon as they were
returned to the holding tank, each of the six fish treated with the anesthesia (four experimental,
one control, and one fish using the ONMNR sampling method) was able to swim upright and
showed no lasting effects. This is rarely seen with MS-222. Field personnel wishing to use MS222 instead of clove oil will have to experiment with varying concentrations to find the
appropriate concentration for the spray bottle application. The advantage of MS-222 over clove
oil is in its solubility in water thereby eliminating the use of Ethanol, which in turn will facilitate
the use of higher concentrations.
One problem was noted in the field tests with regard to anesthesia. The spray bottle had to be
tilted to one side and was awkward to use while the fish was lying on the table. When the
method is used in the field, it is likely that the fish will be lying on the deck of a boat further
complicating the situation. A different spraying device needs to be found. As the method states,
it is important that the spray bottle deliver a fine mist and not a stream.
Our initial chemical analyses were conducted to acquire baseline data and determine the
reproducibility of data in sturgeon given our instrument and conditions. There were no
significant differences seen between consecutive runs of the same tissue samples from Lab Fish
1 or 2. Tests on Lab Fish 3 indicate that testing as little as 1 gram of tissue is not appreciably
different from testing a 10-gram sample.
Conclusion:
The method developed for this study is in fact non-lethal. Furthermore, the results obtained from
the samples can be used to monitor organic contaminants in lake sturgeon but there are two
hurdles to overcome for any such monitoring project to be effective. First, the anesthesia
delivery method must be refined so that it is less cumbersome. Second, better communication
needs to be developed between agencies so that the same fish are not caught and sampled
repeatedly. This would yield misleading or useless data. Each entity that tags fish in the Great
Lakes uses a tag that best suits there own preferences. Using a standardized tag for this
methodology would solve the problem of identifying sampled fish however; researchers would
likely be hesitant to use a tag significantly different from the kind to which they are already
accustomed. A basin-wide database for this information would be most useful. As for the
method, researchers are advised to tag fish but no specific device is indicated. Tag numbers for
this study are given in Table 2 so that these fish can be identified upon subsequent capture.
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Tables, Figures and Appendices:
Figure 1. The location of the liver in a lake sturgeon.
Figure 2. Posterior-to-anterior entrance and exit wounds.
Figure 3. Sampling method employed with no initial incision (top, test fish 1) and with the
incision (middle, test fish 3) along with typical ventral wound (bottom, test fish 4).
Figure 4. Location and size of final sampling site (lab fish).
Figure 5. Wound left by the ONMNR sampling technique.
Figure 6. The relationship between total length and total PCB in muscle tissue from fish
collected in Lake Huron in 2002.
Table 1. Contaminant data from all sampled fish.
Table 2. Physical Parameters of the fish sampled during the field tests.
Appendix A. A Non-Lethal Procedure for the Determination of Organic Contaminant
Concentrations in Lake Sturgeon (Acipenser fulvescens)
Page 12
liver
Figure 1.
Location of liver in a typical sturgeon.
Page 13
Anterior
Posterior
Exit
Entrance
Figure 2.
Posterior-to-anterior entrance and exit wounds.
Page 14
Damaged tissue from slippage
Figure 3. Sampling method employed with no initial incision (top, test fish 1) and
with the incision (middle, test fish 3) along with typical ventral wound
(bottom, test fish 4).
Page 15
Dorsal Fin
Entrance
Exit
Steel Tube
Figure 4.
Location and size of final sampling site (lab fish).
Page 16
Figure 5.
Wound left by the ONMNR sampling technique.
Page 17
800
Total PCB (ng/g)
700
600
500
y = 1.0312e0.0038x
400
R2 = 0.5971
300
200
100
0
900
1100
1300
1500
1700
Total Length (mm)
Figure 6. The relationship between total length and total PCB in muscle tissue from fish collected in Lake Huron in 2002.
Page 18
Sample Identity
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Test Fish 1
Test Fish 2
Test Fish 3
Test Fish 4
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 4
Sacrificed Fish 4
Sacrificed Fish 4
Sample #
Tissue
1999a212001
liver
1999a212001
liver
1999a212003
blood
1999a212003
blood
1999a212002
liver
1999a212002
liver
1999a212004
blood
1999a212004
blood
20010402004 muscle - 1g
20010402004 muscle - 10g
20010402005 testes - tip
20010402006 testes - mid
20010402001 liver - tip
20010402002 liver - mid
20010402003
fat
20020399001
muscle
20020399002
muscle
20020399003
muscle
20020399004
muscle
20020399005
muscle
20020399006
liver
20020399007
eggs
20020399008
muscle
20020399009
testes
20020399026
liver
20020399016
eggs
20020399018
liver
20020399017
muscle
20020399021
muscle
20020399019
testes
20020399020
liver
Units
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
Table 1. Contaminant data from all sampled fish
31+28
33
22
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
19.18
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
17.61 22.40 2.84
25.99
0
8.72
0
0
0
0
0
0
0
0
0
11.52
0
0
46.37 64.23 6.02
18.83
0
0
22.12
0
0
48.01
0
0
26.71 147.93 24.00
52
49 47+48 44
42
41+71
64
40
63
0
0
0
0
0
63.96
0
0
1.90
0
0
0
0
0
83.88
0
0
2.56
0
0
0
0
0
55.70
0
0
1.74
0
0
0
0
0
66.30
0
0
1.78
0
0
0
0
0
70.92
0
0
1.86
0
0
0
0
0
68.16
0
0
2.08
0
0
0
0
0
64.86
0
3.40 1.70
0
0
0
0
0
71.52
0
5.78 2.02
0
0
0
0
0
0
0
0
3.48
0
0
0
0
0
0
0
0
3.08
0
0
0
0
0
0
0
0
0.72
0
0
0
0
0
7.51
5.78
0
3.67
0
0
0
0
0
12.10
0
0
3.38
0
0
0
0
0
12.64
0
0
5.53
0
0
19.10
0
0
59.42
0
0
27.3
0
0
0
0
0
0
0
0
1.05
0
0
0
0
0
6.33
0
0
0.4
0
0
0
0
0
0
0
0
0.2
0
0
0
0
0
0
0
0
0.38
0
0
7.61 15.61
0
4.24
0
0
0.72
8.77
0
21.06
0
0
13.81
0
0
0.53
5.67
0
19.19
17
0
8.14
0
0
1.63
0
0
10.50 13.72
0
0
0
0
1.23
0
0
0
0
0
0
0
0
8.12
7.91
0
36.31 36.28 65.07 19.51 25.29
0
3.31
6.90 2.29 4.36 16.41
0
9.61
0
1.09 3.92
0
0
11.90 17.85 5.76 12.93
0
0
2.37
17.86 10.85 5.00 38.15 3.80 27.80
4
0
6.49
13.62
0
0
0
0
37.86
0
0
1.37
35.61 25.42 18.97 12.76
0
164.71 5.76 3.72 5.05
38.72 18.61 12.01 39.82
0
46.85
0
0
1.20
Page 19
Sample Identity
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 8
Sacrificed Fish 8
Sacrificed Fish 8
Sample #
20020399025
20020399022
20020399023
20020399024
20020399015
20020399027
20020399012
20020399011
20020399010
20020399028
20020399013
20020399014
Table 1. Continued
Tissue
eggs
liver
muscle
muscle
eggs
liver
muscle
eggs
liver
eggs
liver
muscle
Units
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
31+28
33
8.94
9.42
0
0
17.82 44.42
4.82
0
0
0
18.49
0
9.25
0
0
0
19.20
0
16.27
0
28.82
0
0
0
22
1.33
0
0
0
7.25
0
5.39
0
6.16
8.68
0
0
52
3.83
0
12.81
5.42
12.86
10.66
3.86
11.17
0
12.07
12.32
13.25
49 47+48 44
42
41+71
2.05 3.58 38.13
0
14.52
0
0
59.26
0
14.94
5.60 17.30 47.03 5.32 16.06
4.10 2.83 18.89
0
8.58
0
0
24.56
0
7.95
0
6.63 23.93 11.56 7.61
0
6.37 5.78
0
4.04
0
8.29 36.58
0
9.86
0
7.99 15.09
0
4.41
0
10.80 51.03
0
10.83
0
0
50.60
0
13.96
0
5.14 42.19
0
8.78
64
8.84
0
0
0
0
5.81
0
0
0
0
0
0
40
0.85
0
0
1.44
0
0
0
0
0
0
0
0
63
4.17
3.94
3.21
2.14
1.82
2.32
1.79
3.87
1.17
4.63
4.20
4.65
Page 20
Sample Identity
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Test Fish 1
Test Fish 2
Test Fish 3
Test Fish 4
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 4
Sacrificed Fish 4
Sacrificed Fish 4
Sample #
Tissue
1999a212001
liver
1999a212001
liver
1999a212003
blood
1999a212003
blood
1999a212002
liver
1999a212002
liver
1999a212004
blood
1999a212004
blood
20010402004 muscle - 1g
20010402004 muscle - 10g
20010402005 testes - tip
20010402006 testes - mid
20010402001 liver - tip
20010402002 liver - mid
20010402003
fat
20020399001
muscle
20020399002
muscle
20020399003
muscle
20020399004
muscle
20020399005
muscle
20020399006
liver
20020399007
eggs
20020399008
muscle
20020399009
testes
20020399026
liver
20020399016
eggs
20020399018
liver
20020399017
muscle
20020399021
muscle
20020399019
testes
20020399020
liver
Table 1. Continued
Units
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
74
7.70
6.28
6.96
7.10
8.00
9.16
8.02
8.34
5.91
5.24
1.28
6.95
4.23
7.52
40.26
1.00
1.74
1.00
0.74
1.09
1.05
1.76
1.41
8.20
3.67
2.13
2.69
5.03
6.75
26.12
6.86
70+76
20.72
18.46
17.38
15.84
20.14
21.5
19.46
19.00
2.68
1.73
0.65
1.73
1.36
2.24
10.32
1.20
1.54
0.68
0.65
1.25
1.89
3.13
1.12
9.88
3.65
2.21
3.13
4.90
6.97
23.68
7.45
66
95 91 56+60 84+92+89 101
99
119 83
44.72
0
0 13.18
89.62
30.50 27.50 1.10 2.80
38.00
0
0 11.20
75.62
23.68 23.02 1.06 2.40
38.84
0
0 11.06
78.48
27.92 23.44 1.22 2.50
35.14
0
0 10.28
68.30
23.3
19.32 1.60 2.24
46.84
0
0 13.58
87.68
28.66 26.86 1.16 3.40
48.64
0
0 13.66
93.32
31.34 29.66 1.22 3.26
41.86 12.70 0 12.12
81.56
26.26 25.52 0.96 2.60
43.42
0
0 12.30
88.66
27.20 28.02 1.00 2.78
6.43
0
0 5.35
42.48
30.14 18.07 0.56
0
6.69
0
0 3.95
43.84
31.36 17.68 0.62
0
1.41
0
0 0.94
7.34
5.23
3.32 0.16
0
9.26
0
0 5.47
60.17
39.52 23.22 0.67 0.34
6.26
0
0 3.81
42.43
27.64 14.87 0.56 0.36
8.11
0
0 6.02
64.96
41.60 25.38 0.91 0.34
46.38 59.8 0 31.78 368.57 254.05 143.57 4.33 2.19
1.87
0
0 0.78
3.74
2.26
0.89 0.08
0
3.53
0
0 1.09
8.15
5.20
2.57 0.11
0
1.10
0
0
0
2.43
1.30
0.76
0
0
0.95
0
0 0.51
2.26
1.66
0.84
0
0
1.71
0
0 0.61
7.97
5.06
2.04 0.12 0.24
2.48
0
0 0.88
6.29
4.84
1.85 0.09
0
3.85
0
0 1.30
14.99
8.43
4.27 0.17 0.36
2.96
0
0 0.71
6.41
4.28
2.29
0
0
16.76
0
0 6.56
69.6
36.28 17.44 0.56 2.36
7.02
0
0 1.95
23.04
11.71
5.59 0.24 0.60
5.75
0
0 1.88
13.37
8.21
4.56 0.13 0.36
6.57
0
0 2.07
16.97
9.63
4.46 0.26 0.42
12.94
0
0 4.16
34.51
21.98 10.67 0.54 0.85
22.94
0
0 7.02
36.02
22.79 15.71 0.74 1.04
91.71
0
0 25.19 147.61
85.52 69.25 2.68 3.40
24.09
0
0 6.17
36.00
19.37 15.99 0.59 0.57
97
81+87
12.82 44.10
10.78 37.88
8.92 38.30
7.62 34.72
12.78 44.7
12.04 48.22
8.72 42.66
9.48 44.96
0
20.31
0
19.75
0
2.98
0
27.43
0
19.02
0
28.40
2.46 161.47
19.09 10.22
0.66 16.73
0
4.85
0
7.08
0.58
7.95
0.88
8.72
1.26 14.86
0.62 10.12
6.24 15.48
1.79 11.85
0.69
9.22
1.98
9.26
1.86 28.79
1.42 17.21
4.33 90.78
1.50 29.06
Page 21
Sample Identity
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 8
Sacrificed Fish 8
Sacrificed Fish 8
Sample #
20020399025
20020399022
20020399023
20020399024
20020399015
20020399027
20020399012
20020399011
20020399010
20020399028
20020399013
20020399014
Table 1. Continued
Tissue
eggs
liver
muscle
muscle
eggs
liver
muscle
eggs
liver
eggs
liver
muscle
Units
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
74 70+76 66
5.76 2.79 10.06
4.98 3.60 7.24
4.03 2.65 6.81
2.01 1.68 4.53
2.37 2.60 4.57
2.71 2.75 3.98
2.02 0.87 2.26
4.30 2.57 5.77
1.73 1.68 2.65
2.52 2.99 4.73
2.42 3.26 4.68
2.80 2.74 4.46
95
0
0
0
0
0
0
0
0
0
0
0
0
91 56+60 84+92+89 101
0 3.93
55.6
35.09
0 3.16
42.26
27.36
0 2.65
34.62
24.56
0 1.71
25.26
16.31
0 1.65
14.09
11.90
0 1.79
17.54
13.25
0 1.27
18.44
11.83
0 3.55
25.09
19.55
0 1.08
11.18
8.25
0 1.83
16.15
11.62
0 2.16
14.34
11.80
0 1.80
18.00
11.90
99
17.12
13.04
10.68
5.56
4.69
5.16
5.79
9.93
3.26
5.24
4.30
4.17
119
0.62
0.60
0.67
0.25
0.16
0.30
0.19
0.53
0.21
0.20
0.24
0.23
83
0.45
0.70
0.51
0.36
0.41
0.43
0.19
0
0
0.38
0
0.45
97
0.57
0.92
0.96
0.29
1.19
1.70
0.38
0.66
0.51
1.18
1.86
1.43
81+87
24.71
30.16
15.88
12.12
18.11
12.69
8.63
19.54
14.35
13.20
11.50
20.13
Page 22
Sample Identity
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Test Fish 1
Test Fish 2
Test Fish 3
Test Fish 4
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 4
Sacrificed Fish 4
Sacrificed Fish 4
Table 1. Continued
Sample #
Tissue
1999a212001
liver
1999a212001
liver
1999a212003
blood
1999a212003
blood
1999a212002
liver
1999a212002
liver
1999a212004
blood
1999a212004
blood
20010402004 muscle - 1g
20010402004 muscle - 10g
20010402005 testes - tip
20010402006 testes - mid
20010402001 liver - tip
20010402002 liver - mid
20010402003
fat
20020399001
muscle
20020399002
muscle
20020399003
muscle
20020399004
muscle
20020399005
muscle
20020399006
liver
20020399007
eggs
20020399008
muscle
20020399009
testes
20020399026
liver
20020399016
eggs
20020399018
liver
20020399017
muscle
20020399021
muscle
20020399019
testes
20020399020
liver
Units
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
85
7.80
6.88
7.66
6.64
7.76
7.20
7.52
7.30
16.59
6.12
4.45
5.68
4.51
5.23
8.81
0
11.05
0
0
0.83
0.71
1.56
1.31
9.28
2.52
5.26
2.87
7.21
10.07
51.64
11.52
77
2.00
1.68
1.74
1.48
1.84
1.96
1.78
1.70
0
0.13
0
0.11
0.07
0.10
0.53
0
0.09
0
0
0.07
0.15
0.10
0.10
0.76
0.23
0.14
0.19
0.39
0.44
1.86
0.53
110
43.18
35.20
38.56
34.56
42.56
45.50
40.54
41.54
8.64
7.71
1.36
9.89
6.67
11.05
58.72
2.80
4.79
1.33
1.42
4.03
4.71
6.28
3.47
39.44
11.17
7.85
8.46
18.47
22.82
88.24
19.51
82
151
144
107 123
149
118 134 114
4.46 6.26 7.50 8.62 0.66 34.20 47.62 1.50 3.56
3.28 5.04 7.26 7.38 0.54 29.64 42.28 1.28 3.44
4.48 5.80 7.86 8.08 0.56 31.68 44.22 1.42 3.38
3.04 4.86 5.96 6.44 0.46 26.88 35.58 1.30 3.12
3.86 5.66 8.56 8.34 0.66 34.48 49.12 1.44 3.74
3.84 6.66 8.90 9.22 0.64 36.18 53.62 1.46 4.14
4.62 6.18 8.50 9.34 0.56 35.14 51.10 1.50 3.74
4.10 5.78 7.58 9.58 0.62 33.34 48.64 6.88 4.22
0.68 4.97 3.90 3.55 0.06 17.48 4.79 0.45 2.33
0
5.36 4.05 3.68 0.04 14.90 4.76 0.22 2.58
0
0.88 0.68 0.51
0
2.17
0.78
0
0.68
0
7.20 5.79 4.57 0.04 21.13 5.71 0.31 3.04
0
4.95 3.55 3.15 0.03 13.12 4.17 0.14 2.28
0
7.49 5.61 5.17 0.05 20.52 6.81 0.29 3.29
0.32 44.38 33.96 29.47 0.30 129.45 38.46 1.65 18.33
0
0.38 0.51 0.19 0.01 1.28
1.06 0.11 0.66
0.18 1.04 0.98 0.52 0.03 4.06
2.30 0.22 0.88
0
0.27 0.35 0.11 0.01 1.07
0.66
0
0.65
0
0.25 0.27 0.11 0.01 0.88
0.60 0.09 0.36
0.24 1.51 1.10 0.53 0.02 4.24
1.73 0.18 0.71
0.31 0.86 0.75 0.28 0.01 2.82
1.67 0.14 0.61
0.30 2.18 1.69 0.83 0.04 6.82
3.15 0.24 1.03
0.17 0.94 1.00 0.43 0.03 3.88
1.76 0.24 0.80
0
8.68 7.44 3.56 0.20 29.52 16.32 1.40 2.56
0.64 2.67 2.31 1.15 0.06 7.94
5.30 0.37 1.21
0.17 1.48 1.23 1.00 0.04 4.76
4.40 0.25 1.05
0.95 1.71 1.50 1.15 0.04 5.41
5.04 0.30 1.12
0.49 4.08 3.31 2.76 0.11 12.62 11.89 0.62 1.97
1.46 3.33 2.94 4.70 0.13 11.76 19.9 0.48 2.89
6.39 14.67 10.31 20.51 0.92 48.45 90.13 1.57 11.33
1.63 3.28 2.53 4.38 0.24 10.87 20.59 0.47 2.74
Page 23
Sample Identity
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 8
Sacrificed Fish 8
Sacrificed Fish 8
Table 1. Continued
Sample #
20020399025
20020399022
20020399023
20020399024
20020399015
20020399027
20020399012
20020399011
20020399010
20020399028
20020399013
20020399014
Tissue
eggs
liver
muscle
muscle
eggs
liver
muscle
eggs
liver
eggs
liver
muscle
Units
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
85
5.77
20.70
9.58
5.35
9.74
10.74
5.27
9.92
10.25
9.56
19.66
9.44
77
0.21
0.18
0.09
0.17
0.25
0.23
0.10
0.17
0.06
0.16
0.30
0.25
110
13.24
13.12
10.56
12.10
8.84
9.39
4.91
9.90
4.99
10.81
10.86
10.53
82
0.09
0.34
0.34
0.10
0.31
0.27
0.78
0.46
0.21
0.39
0.56
0.60
151
14.90
11.10
10.28
10.73
5.36
6.99
2.29
3.11
1.32
4.13
3.74
4.51
144
7.75
6.24
5.86
5.93
3.38
4.53
1.94
2.65
1.24
0.73
2.40
2.86
107
4.03
3.12
3.18
1.36
0.79
1.13
1.44
2.60
0.96
1.04
0.94
1.02
123
0.12
0.14
0.09
0.08
0.06
0.05
0.04
0.05
0.02
0.04
0.06
0.05
149
34.61
26.76
25.59
22.40
11.07
15.40
7.06
10.29
4.33
10.05
10.14
10.74
118
8.67
6.84
6.44
4.10
2.91
3.49
2.56
4.75
2.09
3.88
3.78
3.93
134
0.47
0.34
0.33
0.40
0.41
0.38
0.15
0.30
0.09
0.52
0.60
0.49
114
2.91
2.22
2.22
1.59
1.03
1.13
1.53
2.02
1.13
0.98
0.92
1.18
Page 24
Sample Identity
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Test Fish 1
Test Fish 2
Test Fish 3
Test Fish 4
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 4
Sacrificed Fish 4
Sacrificed Fish 4
Table 1. Continued
Sample #
Tissue
1999a212001
liver
1999a212001
liver
1999a212003
blood
1999a212003
blood
1999a212002
liver
1999a212002
liver
1999a212004
blood
1999a212004
blood
20010402004 muscle - 1g
20010402004 muscle - 10g
20010402005 testes - tip
20010402006 testes - mid
20010402001
liver - tip
20010402002 liver - mid
20010402003
fat
20020399001
muscle
20020399002
muscle
20020399003
muscle
20020399004
muscle
20020399005
muscle
20020399006
liver
20020399007
eggs
20020399008
muscle
20020399009
testes
20020399026
liver
20020399016
eggs
20020399018
liver
20020399017
muscle
20020399021
muscle
20020399019
testes
20020399020
liver
Units
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
131
3.68
3.24
3.18
2.80
3.90
4.12
3.62
3.62
1.51
1.33
0.22
1.95
1.27
2.12
12.01
0.09
0.27
0.08
0.06
0.25
0.16
0.37
0.18
1.52
0.38
0.34
0.37
0.82
1.21
5.27
1.14
146 153+132 105
141 176+137 138+163 158
22.76 78.48
29.14 12.52
0.40
96.34
6.10
20.36 73.36
25.14 11.48
0.28
88.40
5.16
21.04 73.42
25.32 12.20
0.40
93.92
5.38
17.14 61.68
21.76 10.82
0.36
73.94
4.82
21.90 81.62
28.68 13.46
0
98.78
6.26
25.36 90.22
30.76 14.34
0.48
112.6
7.20
24.50 84.92
30.60 14.72
0.42
107.22 6.54
23.50 82.08
29.26 14.52
0.38
101.38 6.38
8.43
37.36
11.74 11.74
0.44
43.42
5.75
8.95
40.89
12.78 12.80
0.36
48.47
6.13
1.27
5.33
1.79
1.80
0.07
6.09
0.88
12.66 59.05
15.59 18.69
0.64
67.50
9.13
8.24
37.38
10.96 11.53
0.33
43.57
5.91
13.23 59.66
17.26 18.22
0.53
67.76
9.40
76.55 352.28 105.39 112.74
2.90
415.37 53.42
0.58
2.19
0.71
0.82
0.09
2.55
0.25
1.44
6.18
1.67
1.92
0.14
6.92
0.78
0.36
1.39
0.39
0.67
0.05
1.52
0.14
0.28
0.98
0.33
0.51
0.11
1.39
0.12
1.97
7.96
1.29
2.79
0.19
7.74
1.17
1.09
4.30
1.03
1.70
0.13
4.72
0.64
2.81
10.87
2.43
3.96
0.21
12.76
1.58
1.16
4.66
1.11
2.02
0.18
5.89
0.81
11.6
44.56
11.36 15.72
1.12
55.96
6.28
3.15
13.05
3.30
4.52
0.33
15.82
1.65
2.04
8.88
3.24
2.66
0.13
12.09
1.10
2.29
10.08
3.40
3.23
0.16
12.39
1.68
5.60
25.75
8.37
7.76
0.26
31.55
3.50
8.13
29.20
15.96
6.25
0.26
39.17
3.01
41.43 160.62 70.95 31.55
1.36
206.03 17.76
8.09
28.49
15.62
6.61
0.33
39.92
3.37
129
0.54
0.42
0.46
0.48
0.44
0.58
0.56
0.52
0.07
0.05
0
0.04
0.05
0.06
0.40
0
0
0
0
0.01
0
0.01
0
0.04
0.01
0.01
0.01
0.02
0.17
0.19
0.01
126
0.52
0.42
0.44
0.38
0.54
0.58
0.54
0.48
0
0
4.19
0.06
0.03
0.06
0.39
0.02
0.06
0.02
0.02
0.12
0.06
0.14
0.06
0.48
0.17
0.06
0.09
0.21
0.36
2.00
0.38
Page 25
Sample Identity
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 8
Sacrificed Fish 8
Sacrificed Fish 8
Table 1. Continued
Sample #
20020399025
20020399022
20020399023
20020399024
20020399015
20020399027
20020399012
20020399011
20020399010
20020399028
20020399013
20020399014
Tissue
eggs
liver
muscle
muscle
eggs
liver
muscle
eggs
liver
eggs
liver
muscle
Units
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
131
2.09
1.56
1.57
0.92
0.43
0.70
0.59
0.81
0.29
0.41
0.42
0.45
146 153+132 105
19.72 89.65
14.89
15.58 71.12
9.34
14.51 67.13
9.89
8.40
37.81
4.38
4.12
17.49
2.59
5.67
24.59
3.28
4.29
18.66
5.09
5.32
22.22
8.76
2.28
9.18
3.00
3.86
16.45
3.15
3.5
14.18
2.76
4.14
16.89
3.07
141 176+137 138+163 158
18.68
0.59
104.55 9.33
14.92
0.56
73.18
7.46
14.25
0.47
69.11
6.82
13.56
0.81
42.65
4.46
7.29
0.41
22.69
2.32
9.45
0.66
29.19
2.88
5.02
0.20
17.64
2.30
6.97
0.30
26.54
2.97
2.77
0.13
8.97
1.17
6.14
0.29
21.72
2.07
5.44
0.34
18.74
1.64
6.15
0.37
21.43
2.05
129
0.09
0.10
0.07
0.01
0.06
0.08
0.01
0.07
0.04
0.14
0.16
0.08
126
0.79
0.78
0.71
0.46
0.26
0.38
0.15
0.18
0.09
0.24
0.12
0.22
Page 26
Sample Identity
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Test Fish 1
Test Fish 2
Test Fish 3
Test Fish 4
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 4
Sacrificed Fish 4
Sacrificed Fish 4
Sample #
Tissue
1999a212001
liver
1999a212001
liver
1999a212003
blood
1999a212003
blood
1999a212002
liver
1999a212002
liver
1999a212004
blood
1999a212004
blood
20010402004 muscle - 1g
20010402004 muscle - 10g
20010402005 testes - tip
20010402006 testes - mid
20010402001 liver - tip
20010402002 liver - mid
20010402003
fat
20020399001
muscle
20020399002
muscle
20020399003
muscle
20020399004
muscle
20020399005
muscle
20020399006
liver
20020399007
eggs
20020399008
muscle
20020399009
testes
20020399026
liver
20020399016
eggs
20020399018
liver
20020399017
muscle
20020399021
muscle
20020399019
testes
20020399020
liver
Table 1. Continued
Units
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
178
8.34
7.84
7.72
6.82
8.26
10.08
9.04
8.66
2.47
2.99
0.49
4.81
2.79
4.51
26.67
0.24
0.59
0.21
0.15
0.97
0.46
1.19
0.60
4.48
1.34
0.63
0.77
1.93
3.04
17.95
3.00
175 187+182 183 128 167 185 174
177 202 171
156
1.50 22.40 10.54 0.88 4.18 0.46 9.12 10.72 1.76 2.72 2.98
1.36 22.36
9.16 0.92 3.96 0.40 7.84 9.54 1.72 2.70 2.76
1.30 22.70 10.10 0.84 3.82 0.48 8.14 9.32 1.64 2.42 2.70
1.36 18.58
8.48 0.84 3.42 0.46 7.20 8.22 1.36 2.28 2.30
1.38 23.46 10.64 1.04 4.02 0.48 8.78 10.52 1.64 2.84 2.84
1.86 27.28 12.24 1.14 4.60 0.54 9.66 12.16 1.96 3.26 3.52
1.52 25.04 11.56 1.14 4.46 0.50 8.98 10.78 1.94 3.06 3.14
1.50 23.86 11.04 1.22 4.34 0.48 9.00 10.56 1.74 2.88 3.02
0.83
9.28
6.19 0.43 2.40 0.94 5.65 4.18 0.78 1.73 2.87
0.62 10.16
6.81 0.39 2.45 0.85 5.34 4.33 0.93 1.73 3.21
0.11
1.42
1.00 0.06 0.39 0.16 0.86 0.69 0.27 0.25 0.44
1.26 17.27 11.81 0.67 3.37 1.44 9.38 7.42 1.48 2.95 4.49
0.73
9.64
6.47 0.36 2.06 0.81 5.26 3.94 0.83 1.51 2.98
1.11 15.53 10.47 0.58 3.41 1.33 8.92 6.49 1.18 2.30 4.80
6.24 89.25 60.71 3.24 19.13 7.92 45.62 37.96 6.73 14.59 25.28
0.06
0.66
0.40
0
0.15 0.06 0.33 0.39 0.19 0.15 0.19
0.14
1.64
1.15 0.07 0.52 0.17 0.90 0.85 0.26 0.33 0.38
0.09
0.52
0.30
0
0.13 0.03 0.26 0.26 0.17 0.09 0.10
0.07
0.34
0.21
0
0.12 0.02 0.20 0.15 0.13 0.09 0.05
0.28
3.06
1.92 0.12 0.30 0.29 1.89 1.45 0.43 0.49
0.6
0.15
1.59
1.01 0.06 0.17 0.15 0.95 0.73 0.25 0.28 0.07
0.47
3.52
2.13 0.11 0.71 0.30 2.35 1.61 0.42 0.63 0.13
0.19
1.60
1.03 0.06 0.28 0.16 1.08 0.85 0.32 0.38 0.34
1.36 14.16
8.44 0.52 3.12 1.32 11.12 7.12 1.52 2.52
0.6
0.32
3.78
2.26 0.11 0.81 0.40 2.65 1.95 0.44 0.67 0.11
0.18
1.99
1.32 0.08 0.98 0.17 1.07 1.08 0.30 0.44 0.58
0.24
2.45
1.67 0.11 0.82 0.25 1.45 1.38 0.29 0.57 0.11
0.50
5.41
4.03 0.23 2.11 0.48 2.93 2.89 0.57 1.08 0.21
0.61
7.29
3.94 0.21 2.38 0.41 3.71 3.98 0.79 1.40 2.03
3.27 46.02 25.64 1.40 11.16 2.57 23.07 24.88 4.99 8.13 13.26
0.58
7.31
3.93 0.22 2.25 0.40 3.74 4.09 0.74 1.33 2.74
Page 27
Sample Identity
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 8
Sacrificed Fish 8
Sacrificed Fish 8
Sample #
20020399025
20020399022
20020399023
20020399024
20020399015
20020399027
20020399012
20020399011
20020399010
20020399028
20020399013
20020399014
Table 1. Continued
Tissue
eggs
liver
muscle
muscle
eggs
liver
muscle
eggs
liver
eggs
liver
muscle
Units
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
178
7.58
7.22
6.68
4.70
2.13
3.18
1.64
1.66
0.77
1.87
1.74
1.93
175 187+182 183 128
1.51 40.96 14.68 0.77
1.66 35.74 13.90 0.76
1.37 35.49 12.43 0.70
1.05 22.30
8.99 0.42
0.53
9.28
4.14 0.23
0.78 14.06
6.00 0.38
0.35
5.22
3.29 0.19
0.65
5.60
3.45 0.17
0.29
2.65
1.60 0.09
0.46
0.21
3.48 0.19
0.52
7.74
2.94 0.16
0.52 10.25
3.80 0.19
167
4.40
2.56
2.37
1.41
0.95
1.03
0.74
1.42
0.43
1.00
0.80
0.88
185 174
177 202
2.11 9.97 11.40 1.37
1.90 8.64 10.58 1.66
1.74 7.98 10.07 1.39
1.47 10.18 7.63 1.00
0.69 5.04 3.57 0.51
1.00 6.85 5.09 0.66
0.42 2.53 2.37 0.56
0.45 2.69 2.62 0.57
0.21 1.34 1.21 0.37
0.55 4.01 3.00 0.53
0.40 3.50 2.56 0.44
0.58 4.05 3.04 0.55
171
4.39
4.26
3.94
2.73
1.27
1.94
0.89
0.90
0.48
1.13
0.94
1.29
156
3.35
3.12
3.09
1.50
0.22
0.37
1.38
1.90
0.82
0.20
0.20
0.90
Page 28
Sample Identity
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Test Fish 1
Test Fish 2
Test Fish 3
Test Fish 4
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 4
Sacrificed Fish 4
Sacrificed Fish 4
Table 1. Continued
Sample #
Tissue
1999a212001
liver
1999a212001
liver
1999a212003
blood
1999a212003
blood
1999a212002
liver
1999a212002
liver
1999a212004
blood
1999a212004
blood
20010402004 muscle - 1g
20010402004 muscle - 10g
20010402005 testes - tip
20010402006 testes - mid
20010402001
liver - tip
20010402002 liver - mid
20010402003
fat
20020399001
muscle
20020399002
muscle
20020399003
muscle
20020399004
muscle
20020399005
muscle
20020399006
liver
20020399007
eggs
20020399008
muscle
20020399009
testes
20020399026
liver
20020399016
eggs
20020399018
liver
20020399017
muscle
20020399021
muscle
20020399019
testes
20020399020
liver
Units
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
173 157 200 172 197
180
193 191 199 170+190 198
0.10 2.16 0.92 4.88 0.52 29.16 3.18 0.78 0.34 10.28 0.38
0.08 1.92 0.66 4.34 0.46 26.48 2.94 0.62 0.28
9.04
0.34
0.08 1.98 0.74 4.64 0.44 27.74 3.06 0.66 0.34
9.40
0.44
0.10 1.68 0.64 3.76 0.44 22.94 2.56 0.58 0.32
8.20
0.28
0.10 2.06 0.68 4.62 0.54 28.96 3.38 0.72 0.32
9.74
0.34
0.12 2.50 0.84 5.38 0.50 33.74 3.64 0.76 0.38 11.60 0.42
0.10 2.26 0.86 5.00 0.58 31.02 3.26 0.78 0.34 10.38 0.38
0.12 2.14 0.86 5.12 0.56 30.48 3.60 0.80 0.36 10.26 0.36
0
1.06 0.45 3.03 0.38 21.60 1.73 0.63 0.45
8.41
0.34
0.02 1.42 0.53 3.19 0.38 22.98 1.86 0.58 0.38
9.01
0.29
0
0.17 0.11 0.51 0.05 3.23 0.28 0.10 0.06
1.22
0.06
0.02 1.90 1.12 5.76 0.81 42.39 3.34 1.12 0.83 15.69 0.52
0.02 1.41 0.56 3.24 0.35 23.46 1.78 0.60 0.39
8.56
0.30
0.05 1.81 0.93 5.28 0.54 37.77 2.76 1.06 0.59 13.51 0.38
0.23 10.97 5.60 28.72 3.34 208.42 16.20 5.20 3.70 74.01 2.33
0
0.05 0.05 0.25 0.08 1.15 0.12 0.05 0.04
0.60
0.04
0.02 0.10 0.09 0.52 0.08 3.11 0.30 0.13 0.10
1.82
0.05
0
0.06 0.06 0.21 0.09 0.76 0.08 0.04 0.05
0.39
0.03
0.01 0.05 0.06 0.13
0
0.49 0.05 0.04 0.05
0.29
0.02
0.11 0.17 0.38 1.16 0.27 5.94 0.52 0.17 0.23
2.56
0.15
0.07 0.13 0.15 0.56 0.10 3.00 0.25 0.07 0.12
1.26
0.06
8.44 0.20 0.31 1.09 0.19 6.40 0.55 0.20 0.21
3.11
0.12
0
0.08 0.11 0.63 0.14 2.59 0.27 0.08 0.13
1.24
0.06
0.24 0.88 1.28 4.96 1.12 26.28 2.32 0.56 1.00 11.56 0.52
0.05 0.24 0.29 1.30 0.13 6.41 0.57 0.15 0.26
2.79
0.14
0.02 0.16 0.15 0.63 0.08 3.75 0.29 0.13 0.09
1.93
0.08
0.03 0.24 0.08 0.93 0.13 4.98 0.41 0.13 0.12
2.53
0.07
0.06 0.51 0.33 2.04 0.30 12.36 1.00 0.09 0.25
6.38
0.16
0.08 0.64 0.37 2.97 0.33 15.26 1.56 0.58 0.38
7.05
0.24
1.04 3.36 3.80 20.69 3.02 99.53 9.21 1.93 2.23 45.01 1.94
0.07 0.67 0.44 3.02 0.34 15.18 1.64 0.30 0.28
6.96
0.27
Page 29
Sample Identity
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 8
Sacrificed Fish 8
Sacrificed Fish 8
Table 1. Continued
Sample #
20020399025
20020399022
20020399023
20020399024
20020399015
20020399027
20020399012
20020399011
20020399010
20020399028
20020399013
20020399014
Tissue
eggs
liver
muscle
muscle
eggs
liver
muscle
eggs
liver
eggs
liver
muscle
Units
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
173
0.20
0.04
0.33
0.08
0.08
0.04
0.12
0.09
0.05
0.09
0.08
0.08
157
0.72
0.94
0.81
0.31
0.95
0.92
0.30
0.92
0.95
0.31
0.30
0.32
200
1.10
1.34
1.11
0.86
0.26
0.46
0.34
0.31
0.14
0.50
0.34
0.36
172
5.16
6.72
6.11
3.53
1.94
2.67
1.81
2.01
0.99
1.88
1.26
1.63
197
0.55
0.68
0.71
0.52
0.20
0.30
0.25
0.26
0.14
0.26
0.18
0.28
180
38.96
41.98
41.45
27.65
12.97
20.58
11.35
11.59
6.04
12.46
10.04
13.40
193
3.75
3.90
3.41
2.19
1.10
1.59
0.84
0.95
0.47
1.10
0.80
1.13
191
1.03
1.38
1.18
0.85
0.45
0.87
0.27
0.39
0.19
0.32
0.34
0.45
199 170+190 198
0.50 16.69 0.42
0.60 17.98 0.48
0.58 16.09 0.52
0.72 11.29 0.33
0.41
5.13
0.16
0.52
8.38
0.24
0.24
4.52
0.17
0.23
5.29
0.13
0.13
2.49
0.10
0.33
5.21
0.15
0.26
4.44
0.14
0.33
5.53
0.17
Page 30
Sample Identity
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Test Fish 1
Test Fish 2
Test Fish 3
Test Fish 4
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 4
Sacrificed Fish 4
Sacrificed Fish 4
Table 1. Continued
Sample #
Tissue
1999a212001
liver
1999a212001
liver
1999a212003
blood
1999a212003
blood
1999a212002
liver
1999a212002
liver
1999a212004
blood
1999a212004
blood
20010402004 muscle - 1g
20010402004 muscle - 10g
20010402005 testes - tip
20010402006 testes - mid
20010402001
liver - tip
20010402002 liver - mid
20010402003
fat
20020399001
muscle
20020399002
muscle
20020399003
muscle
20020399004
muscle
20020399005
muscle
20020399006
liver
20020399007
eggs
20020399008
muscle
20020399009
testes
20020399026
liver
20020399016
eggs
20020399018
liver
20020399017
muscle
20020399021
muscle
20020399019
testes
20020399020
liver
Units
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
201
8.64
7.82
8.72
6.92
8.42
9.84
8.78
8.90
6.94
6.92
1.00
14.46
7.37
11.25
65.02
0.56
1.11
0.46
0.33
3.34
1.40
2.91
1.50
13.12
2.95
1.29
1.65
4.02
6.46
49.81
5.78
203
10.32
9.10
9.28
8.18
9.56
11.88
10.96
10.36
8.81
8.86
1.39
18.35
8.95
14.32
84.44
0.64
1.29
0.44
0.34
3.70
1.64
3.00
1.61
13.4
3.06
1.52
2.14
5.31
6.13
54.39
6.11
Total 189 195 208 207 1984 205 206
209
PCB
0.68 1.32 0.70 0.52 3.30 0.26 1.78 0.86 935.14
0.62 1.16 0.62 0.46 3.00 0.22 1.52 0.76 849.98
0.56 1.18 0.60 0.44 2.88 0.26 1.56 0.74 850.58
0.52 1.06 0.52 0.38 2.52 0.18 1.26 0.62 743.02
0.62 1.28 0.62 0.50 3.04 0.24 1.52 0.80 944.14
0.72 1.52 0.82 0.60 3.74 0.32 2.00 1.08 1028.52
0.70 1.38 0.68 0.52 3.44 0.28 1.72 0.88 981.20
0.62 1.38 0.64 0.48 3.28 0.28 1.60 0.86 959.24
0.72 1.16 0.77 0.75 4.69 0.37 2.25 2.42 424.01
0.72 1.20 0.69 0.74 4.62 0.42 2.45 2.55 424.84
0.09 0.18 0.15 0.11 0.64 0.05 0.36 0.35
73.42
1.12 2.45 1.29 1.37 9.14 0.65 4.77 4.20 638.17
0.75 1.13 0.67 0.71 4.61 0.35 2.35 2.33 405.22
1.06 1.73 1.02 1.01 7.40 0.52 3.72 3.62 628.56
5.16 10.37 6.02 6.52 39.30 2.73 20.74 20.67 3689.85
0.05 0.13 0.11 0.11 0.31 0.06 0.22 0.31
64.16
0.11 0.27 0.12 0.09 0.90 0.05 0.36 0.20 111.34
0.06 0.08 0.09 0.08 0.26 0.05 0.14 0.14
27.15
0.04 0.08 0.07 0.06 0.18
0
0.14 0.21
27.01
0.24 0.69 0.79 0.96 2.09 0.17 2.06 5.05 138.03
0.17 0.28 0.32 0.35 0.89 0.08 0.69 1.34 161.64
0.24 0.53 0.51 0.55 1.44 0.13 1.13 2.00 246.89
0.15 0.29 0.28 0.27 0.74 0.09 0.63 1.15 104.52
1.24 2.56 1.88 2.28 5.60 0.60 5.28 8.24 626.71
0.22 0.51 0.42 0.51 1.37 0.12 1.16 1.93 382.18
0.14 0.27 0.16 0.16 0.76 0.09 0.43 0.53 184.10
0.27 0.34 0.23 0.20 1.24 0.12 0.61 0.61 314.45
0.54 0.84 0.52 0.62 3.08 0.25 1.96 2.67 472.44
0.60 1.11 0.60 0.61 3.02 0.32 1.61 1.16 479.12
4.10 9.58 4.94 5.28 26.87 2.38 16.88 10.82 2332.00
0.70 1.12 0.49 0.53 2.86 0.30 1.58 0.83 765.56
Page 31
Sample Identity
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 8
Sacrificed Fish 8
Sacrificed Fish 8
Table 1. Continued
Sample #
20020399025
20020399022
20020399023
20020399024
20020399015
20020399027
20020399012
20020399011
20020399010
20020399028
20020399013
20020399014
Tissue
eggs
liver
muscle
muscle
eggs
liver
muscle
eggs
liver
eggs
liver
muscle
Units 201
203 189
ng/g 12.28 13.17 0.90
ng/g 14.22 17.24 1.60
ng/g 15.24 16.41 1.29
ng/g 9.70 10.07 0.60
ng/g 4.40 3.94 0.41
ng/g 6.19 6.08 0.54
ng/g 4.70 5.70 0.36
ng/g 4.00 4.46 0.50
ng/g 2.40 2.77 0.31
ng/g 4.27 4.27 0.41
ng/g 3.34 3.14 0.34
ng/g 4.71 5.10 0.47
195
2.59
2.92
3.08
1.83
0.75
1.05
0.78
0.66
0.43
0.76
0.64
0.84
208
0.43
0.66
0.77
0.54
0.27
0.37
0.45
0.36
0.29
0.45
0.32
0.50
207
0.46
0.78
0.86
0.57
0.24
0.29
0.54
0.33
0.26
0.55
0.28
0.63
1984
5.34
8.34
8.29
4.39
1.71
2.77
3.03
2.23
1.49
2.11
1.58
2.51
205
0.52
0.92
0.74
0.36
0.17
0.22
0.27
0.24
0.13
0.14
0.20
0.29
206
1.71
3.00
3.37
1.62
0.66
0.87
1.66
1.06
0.87
1.24
0.76
1.40
209
1.06
2.16
3.02
1.50
0.60
0.66
1.84
1.07
0.86
1.92
1.02
2.63
Total PCB
815.41
717.71
745.36
438.74
283.32
376.87
229.29
340.81
185.27
326.47
320.00
313.30
Page 32
Sample Identity
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Test Fish 1
Test Fish 2
Test Fish 3
Test Fish 4
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 4
Sacrificed Fish 4
Sacrificed Fish 4
Sample #
1999a212001
1999a212001
1999a212003
1999a212003
1999a212002
1999a212002
1999a212004
1999a212004
20010402004
20010402004
20010402005
20010402006
20010402001
20010402002
20010402003
20020399001
20020399002
20020399003
20020399004
20020399005
20020399006
20020399007
20020399008
20020399009
20020399026
20020399016
20020399018
20020399017
20020399021
20020399019
20020399020
Table 1. Continued
Tissue
liver
liver
blood
blood
liver
liver
blood
blood
muscle - 1g
muscle - 10g
testes - tip
testes - mid
liver - tip
liver - mid
fat
muscle
muscle
muscle
muscle
muscle
liver
eggs
muscle
testes
liver
eggs
liver
muscle
muscle
testes
liver
PentaalphaHexaOctaUnits chlorobenzene HCH chlorobenzene PCPME Lindane Aldrin Dachtal chlorostyrene
ng/g
0.05
0
1.08
0.16
0.16
0
0.09
0
ng/g
0.06
0
0.90
0.15
0.13
0
0.06
0
ng/g
0.04
0
0.19
0.03
0
0
0.02
0
ng/g
0.04
0
0.26
0.03
0
0
0
0
ng/g
0.04
0
0.78
0.08
0.09
0
0.04
0.69
ng/g
0.03
0
0.64
0.07
0.09
0
0.04
0.67
ng/g
0.01
0
0
0.01
0
0
0
0
ng/g
0.01
0
0
0.01
0
0
0
0
ng/g
0.55
0.64
9.40
0.36
0
0
0.29
46.03
ng/g
0.55
0.47
8.88
0.35
0
0
0.36
53.01
ng/g
0.06
0
1.42
0.06
0
0
0.07
7.61
ng/g
0.65
0.41
10.26
0.38
0.28
0
0.29
66.28
ng/g
0.48
0.32
8.61
0.37
0.27
0
0.21
50.92
ng/g
0.68
0.46
12.81
0.52
0.32
0
0.36
74.94
ng/g
4.35
2.35
83.95
3.25
1.18
0
1.56
486.08
ng/g
0.18
0.12
3.27
0.08
0
0
0.25
0
ng/g
0.14
0.14
2.84
0.12
0
0
0.25
0
ng/g
0.03
0
0.33
0.03
0
0
0.02
0
ng/g
0.02
0.05
0.75
0.02
0
0
0.05
0
ng/g
0.18
0.09
2.81
0.12
0
0
0.20
0
ng/g
0.17
0.07
2.72
0.18
0.10
0
0.14
0
ng/g
0.30
0.18
6.59
0.27
0.16
0
0.41
0
ng/g
0.40
0.26
7.88
0.31
0.23
0
0.42
0
ng/g
2.40
1.08
63.60
2.44
1.56
0
3.08
80.80
ng/g
1.13
0.51
23.18
0.86
0.53
0
1.10
0
ng/g
0.61
0.18
18.17
0.22
0.13
0
0.31
20.50
ng/g
0.59
0.20
18.83
0.24
0.20
0
0.30
0
ng/g
1.73
0.47
54.28
0.60
0.36
0
0.72
0
ng/g
0.17
0.19
3.94
0.24
0
0
0.25
0.83
ng/g
0.33
0.70
13.26
0.71
0.55
0
1.14
2.94
ng/g
0.19
0.25
3.97
0.27
0
0
0.26
0.93
Page 33
Sample Identity
Sample #
Sacrificed Fish 5 200203990250
Sacrificed Fish 5 20020399022
Sacrificed Fish 5 20020399023
Sacrificed Fish 6 20020399024
Sacrificed Fish 6 20020399015
Sacrificed Fish 6 20020399027
Sacrificed Fish 7 20020399012
Sacrificed Fish 7 20020399011
Sacrificed Fish 7 20020399010
Sacrificed Fish 8 20020399028
Sacrificed Fish 8 20020399013
Sacrificed Fish 8 20020399014
Table 1. Continued
Tissue
eggs
liver
muscle
muscle
eggs
liver
muscle
eggs
liver
eggs
liver
muscle
PentaalphaHexaOctaUnits chlorobenzene HCH chlorobenzene PCPME Lindane Aldrin Dachtal chlorostyrene
ng/g
0.99
0.24
26.14
0.39
0.20
0
0.39
22.67
ng/g
0.54
0.16
15.38
0.36
0.16
0
0.36
16.82
ng/g
0.56
0.12
14.15
0.28
0
0
0.32
11.70
ng/g
0.33
0.22
5.36
0.29
0.16
0
0.43
13.66
ng/g
0.27
0.12
4.28
0.25
0
0
0.25
0
ng/g
0.26
0.20
4.43
0.29
0.16
0
0.30
0
ng/g
0.07
0.06
3.41
0.10
0
0
0.20
9.05
ng/g
0.33
0.18
9.35
0.25
0
0
0.36
0
ng/g
0.12
0.07
2.83
0.09
0
0
0.12
0
ng/g
0.67
0.30
12.12
0.34
0
0
0.75
0
ng/g
0.42
0.16
9.26
0.32
0.20
0
0.58
0
ng/g
0.62
0.23
12.24
0.33
0.19
0
0.71
0
Page 34
Sample Identity
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Test Fish 1
Test Fish 2
Test Fish 3
Test Fish 4
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 4
Sacrificed Fish 4
Sacrificed Fish 4
Table 1. Continued
Sample #
1999a212001
1999a212001
1999a212003
1999a212003
1999a212002
1999a212002
1999a212004
1999a212004
20010402004
20010402004
20010402005
20010402006
20010402001
20010402002
20010402003
20020399001
20020399002
20020399003
20020399004
20020399005
20020399006
20020399007
20020399008
20020399009
20020399026
20020399016
20020399018
20020399017
20020399021
20020399019
20020399020
Tissue
liver
liver
blood
blood
liver
liver
blood
blood
muscle - 1g
muscle - 10g
testes - tip
testes - mid
liver - tip
liver - mid
fat
muscle
muscle
muscle
muscle
muscle
liver
eggs
muscle
testes
liver
eggs
liver
muscle
muscle
testes
liver
bHeptachlor Oxytrans(G)- cis(A)- trans(G)- cis(A)Units Epoxide chlordane Chlordane ChlordaneNonachlor Nonachlor
ng/g
0
0.88
0.57
1.96
9.38
0.81
ng/g
0
0.79
0.65
1.87
9.25
0.94
ng/g
0
0.14
0.12
0.25
1.59
0
ng/g
0
0.19
0.08
0.26
1.50
0
ng/g
0
5.79
7.90
20.67
85.23
1.48
ng/g
0.22
5.84
7.59
19.82
83.61
1.34
ng/g
0
0
0.12
0.29
1.36
0
ng/g
0
0
0.14
0.36
1.29
0
ng/g
1.15
1.28
1.03
2.41
11.27
3.87
ng/g
1.09
1.63
1.30
2.76
15.58
6.87
ng/g
0.31
0.22
0.16
0.35
2.56
0.52
ng/g
2.44
1.73
1.32
3.17
16.65
4.09
ng/g
1.80
1.41
1.08
2.55
12.52
3.92
ng/g
2.85
1.95
1.62
3.76
18.32
5.66
ng/g
17.08
9.89
7.53
18.13
91.97
23.93
ng/g
0
0.24
0.12
0.22
0.65
0.39
ng/g
0
0.71
0.32
0.73
2.81
1.30
ng/g
0
0
0.05
0.10
0.33
0.15
ng/g
0
0.07
0.04
0.11
0.35
0.15
ng/g
0.49
0.38
0.60
1.13
4.47
1.99
ng/g
0.16
0.23
0.32
0.58
2.16
0.94
ng/g
0.52
0.80
0.93
1.72
5.35
2.43
ng/g
0.81
0.49
0.68
1.24
3.28
1.53
ng/g
4.72
4.40
5.56
9.52
27.36
12.44
ng/g
1.02
1.58
1.82
3.30
9.31
4.31
ng/g
0.95
0.87
0.45
1.03
2.94
1.44
ng/g
1.13
1.11
0.54
1.07
3.60
1.78
ng/g
2.34
3.15
1.57
3.25
10.40
4.79
ng/g
1.30
2.95
1.69
4.83
15.25
7.77
ng/g
4.05
12.94
6.94
19.13
66.64
34.36
ng/g
1.23
3.03
1.51
4.34
13.37
7.07
Dieldrin Endrin
155.34 1.08
158.33 0.80
22.30
0
22.80
0
2180.41 7.52
2126.75 8.17
26.14
0
27.76
0
63.48
0
80.07
0.52
9.51
0
90.30
0.75
71.24
0.56
109.95 1.01
629.99 4.14
0.89
0
2.80
0
0.39
0
0.47
0
2.24
0
1.64
0
4.04
0.14
4.21
0
26.40
0
14.94
0.55
3.98
0.30
6.84
0
17.81
0.61
11.14
1.28
35.88
4.41
13.59
1.10
Page 35
Sample Identity
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 8
Sacrificed Fish 8
Sacrificed Fish 8
Table 1. Continued
Sample #
20020399025
20020399022
20020399023
20020399024
20020399015
20020399027
20020399012
20020399011
20020399010
20020399028
20020399013
20020399014
Tissue
eggs
liver
muscle
muscle
eggs
liver
muscle
eggs
liver
eggs
liver
muscle
bHeptachlor Oxytrans(G)- cis(A)- trans(G)- cis(A)Units Epoxide chlordane Chlordane ChlordaneNonachlor Nonachlor Dieldrin Endrin
ng/g
0.93
1.57
0.78
2.05
10.25
4.92
4.30
0.34
ng/g
0.78
0.96
0.70
1.58
10.14
4.68
5.08
0
ng/g
0.68
1.14
0.57
1.44
9.64
4.48
3.21
0
ng/g
1.19
1.16
1.56
2.80
8.63
3.88
5.91
0.39
ng/g
0.23
0.61
0.84
1.45
3.83
1.80
3.93
0
ng/g
0.41
0.73
1.06
1.84
5.77
2.49
6.81
0
ng/g
0.41
0.53
0.39
1.03
4.82
2.33
1.68
0.14
ng/g
0.88
0.70
0.65
1.35
5.91
2.64
5.37
0.20
ng/g
0.17
0.37
0.20
0.47
2.24
1.22
1.68
0
ng/g
1.12
0.81
1.13
1.85
5.42
2.56
7.20
0.11
ng/g
1.16
1.02
1.00
1.70
5.08
2.42
6.02
0
ng/g
1.03
1.15
1.29
2.16
6.15
2.87
6.75
0.28
Page 36
Sample Identity
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Test Fish 1
Test Fish 2
Test Fish 3
Test Fish 4
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 4
Sacrificed Fish 4
Sacrificed Fish 4
Table 1 Continued
Sample #
1999a212001
1999a212001
1999a212003
1999a212003
1999a212002
1999a212002
1999a212004
1999a212004
20010402004
20010402004
20010402005
20010402006
20010402001
20010402002
20010402003
20020399001
20020399002
20020399003
20020399004
20020399005
20020399006
20020399007
20020399008
20020399009
20020399026
20020399016
20020399018
20020399017
20020399021
20020399019
20020399020
Tissue
liver
liver
blood
blood
liver
liver
blood
blood
muscle - 1g
muscle - 10g
testes - tip
testes - mid
liver - tip
liver - mid
fat
muscle
muscle
muscle
muscle
muscle
liver
eggs
muscle
testes
liver
eggs
liver
muscle
muscle
testes
liver
Units p,p'-DDE p,p'-DDD p,p'-DDT
ng/g
4.66
9.40
0
ng/g
4.28
8.81
0
ng/g
0.86
0
0
ng/g
0.76
0
0
ng/g 41.19
176.85
273.16
ng/g 40.60
178.50
252.12
ng/g
0.57
2.41
0
ng/g
0.63
0
0
ng/g
3.90
0
0
ng/g
5.23
4.22
0
ng/g
0.57
0
0
ng/g
5.81
5.63
0
ng/g
4.73
4.24
0
ng/g
6.68
4.48
0
ng/g 27.67
23.69
0
ng/g
3.00
0
0
ng/g 11.27
0
0
ng/g
1.79
0
0
ng/g
1.28
0
0
ng/g 18.70
4.58
0
ng/g
9.20
2.55
0
ng/g 28.92
4.41
0
ng/g 12.88
3.94
0
ng/g 109.52
30.44
0
ng/g 33.80
9.42
0
ng/g 15.04
2.81
0
ng/g 15.11
2.80
0
ng/g 40.73
10.44
0
ng/g 73.34
9.36
0
ng/g 314.53
35.04
54.79
ng/g 63.53
10.16
0
photoMirex Mirex
0
0
1.29
0
0
0
0
0
38.63 3.75
37.23 4.36
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
6.42
0
0
0
0
0
0
0
0
0.26
0
0
0
0
0
0
0
1.88
0
0.37
0
0
0
0
0
0.77
0
1.06 0.95
5.77 4.19
1.25
0
Page 37
Sample Identity
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 8
Sacrificed Fish 8
Sacrificed Fish 8
Table 1 Continued
Sample #
20020399025
20020399022
20020399023
20020399024
20020399015
20020399027
20020399012
20020399011
20020399010
20020399028
20020399013
20020399014
Tissue
eggs
liver
muscle
muscle
eggs
liver
muscle
eggs
liver
eggs
liver
muscle
photoUnits p,p'-DDE p,p'-DDD p,p'-DDT Mirex Mirex
ng/g 65.85
7.30
0
6.07 1.33
ng/g 51.06
8.00
0
0
0
ng/g 47.32
6.95
0
0.36 1.09
ng/g 35.98
8.47
0
4.04 0.91
ng/g 18.10
4.21
0
0
0
ng/g 25.61
6.05
0
0
0
ng/g 34.00
2.71
0
0.40 0.86
ng/g 52.48
4.46
0
0.48
0
ng/g 19.07
2.22
0
0
0
ng/g 21.74
7.24
0
0
0
ng/g 20.40
8.14
0
0
0
ng/g 23.89
7.32
0
0
0
Page 38
Sample Identity
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 1
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 2
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Lab Fish 3
Test Fish 1
Test Fish 2
Test Fish 3
Test Fish 4
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 1
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 2
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 3
Sacrificed Fish 4
Sacrificed Fish 4
Sacrificed Fish 4
Table 1. Continued
Sample #
1999a212001
1999a212001
1999a212003
1999a212003
1999a212002
1999a212002
1999a212004
1999a212004
20010402004
20010402004
20010402005
20010402006
20010402001
20010402002
20010402003
20020399001
20020399002
20020399003
20020399004
20020399005
20020399006
20020399007
20020399008
20020399009
20020399026
20020399016
20020399018
20020399017
20020399021
20020399019
20020399020
Tissue
liver
liver
blood
blood
liver
liver
blood
blood
muscle - 1g
muscle - 10g
testes - tip
testes - mid
liver - tip
liver - mid
fat
muscle
muscle
muscle
muscle
muscle
liver
eggs
muscle
testes
liver
eggs
liver
muscle
muscle
testes
liver
Hexa Units Toxaphene
ng/g
0
ng/g
0.78
ng/g
0
ng/g
0.44
ng/g
39.79
ng/g
32.28
ng/g
0
ng/g
0
ng/g
0
ng/g
2.23
ng/g
1.53
ng/g
12.62
ng/g
1.11
ng/g
0.61
ng/g
0
ng/g
0.91
ng/g
1.08
ng/g
0
ng/g
0
ng/g
2.32
ng/g
1.62
ng/g
5.46
ng/g
7.56
ng/g
28.01
ng/g
11.36
ng/g
3.25
ng/g
8.97
ng/g
15.87
ng/g
8.77
ng/g
58.99
ng/g
14.46
Hepta Toxaphene
8.26
5.76
0.35
0.57
35.44
60.92
0
0
17.66
29.36
4.54
0
0
0.92
0
1.90
6.83
0.39
1.47
10.38
5.98
12.86
13.31
126.79
43.44
19.83
32.74
79.00
50.84
340.80
89.39
Octa Toxaphene
13.75
16.89
5.50
2.19
172.89
218.09
0
0
24.36
38.66
4.48
12.23
2.28
2.99
0
1.43
6.94
0.58
0.74
20.27
10.40
19.56
11.63
133.19
44.68
13.42
30.90
73.08
66.49
534.51
159.54
Nona Toxaphene
15.89
15.93
1.14
2.52
145.05
166.63
1.39
0.20
11.80
21.04
2.71
0
0
0
0
0.26
2.60
0.21
0.22
4.54
1.50
5.79
2.70
29.78
12.25
4.19
7.10
15.18
26.34
169.71
43.92
Deca Total Toxaphene Toxaphene
0
37.90
0
39.36
0
6.99
0
5.72
14.39
407.56
11.59
489.51
0
1.39
0
0.20
0
53.82
0
91.29
0
13.25
378.52
403.38
0
3.39
0
4.52
0
0
0
4.49
0
17.44
0
1.19
0
2.43
1.09
38.60
0
19.50
0.19
43.85
0
35.20
2.24
320
0.28
112.02
0
40.69
0.09
79.81
1.06
184.19
0.69
153.00
8.52
1112.53
0.98
308.29
Page 39
Sample Identity
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 5
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 6
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 7
Sacrificed Fish 8
Sacrificed Fish 8
Sacrificed Fish 8
Table 1. Continued
Sample #
20020399025
20020399022
20020399023
20020399024
20020399015
20020399027
20020399012
20020399011
20020399010
20020399028
20020399013
20020399014
Tissue
eggs
liver
muscle
muscle
eggs
liver
muscle
eggs
liver
eggs
liver
muscle
Hexa Units Toxaphene
ng/g
16.16
ng/g
2.75
ng/g
2.17
ng/g
7.13
ng/g
1.66
ng/g
2.97
ng/g
3.14
ng/g
3.71
ng/g
1.00
ng/g
3.90
ng/g
2.19
ng/g
3.76
Hepta Toxaphene
23.65
12.33
11.30
28.96
11.77
15.91
10.09
12.57
3.90
20.26
19.46
23.21
Octa Toxaphene
19.57
11.65
11.34
28.50
11.63
16.18
11.00
11.89
4.01
16.99
14.03
19.55
Nona Toxaphene
5.22
4.28
3.89
6.50
2.73
4.08
3.85
3.84
2.26
4.34
3.85
4.66
Deca Toxaphene
0.28
0
0
0
0
0
0.40
0
0
0
0
0
Total
64.87
31.01
28.70
71.09
27.79
39.13
28.48
32.01
11.16
45.49
39.54
51.17
Page 40
Description
Lab Fish 1
Lab Fish 2
Lab Fish 3
Control for Anesthesia
Control - No Treatment
Test Fish 1
Test Fish 2
Test Fish 3
Test Fish 4
Sacrificed Fish 1
Sacrificed Fish 2
Sacrificed Fish 3
Sacrificed Fish 4
Sacrificed Fish 5
Sacrificed Fish 6
Sacrificed Fish 7
Sacrificed Fish 8
Sex Weight (g) Total Length (mm) Fork Length (mm)
f
1626
1492
m
1588
1416
m
1417
*
7598
1132
1053
*
6917
1055
975
*
4763
935
875
*
9525
1194
1106
*
9639
1170
1093
*
4196
931
870
f
35947
1598
1485
m
16556
1315
1236
f
15876
1323
1219
m
1462
1380
f
25061
1548
1429
f
27329
1476
1403
f
20298
1440
1416
f
24494
1512
1397
Commercial Length**
(mm)
1149
1111
654
599
544
692
651
519
970
810
784
905
943
919
867
908
* Genetic analysis incomplete at this time.
** Commercial Length is measured from the posterior of the operculum to the posterior of the dorsal fin.
Table 2.
Physical Parameters of the fish sampled during the field tests.
Tissue sampled (g) Floy Tag Number
n/a
n/a
n/a
n/a
n/a
n/a
n/a
13254
n/a
13255
1.9
13257
2.5
13259
2.9
13260
1.9
13261
n/a
n/a
n/a
n/a
n/a
n/a
n/a
n/a
n/a
n/a
n/a
n/a
n/a
n/a
n/a
n/a
Page 41
Appendix A
Page 42
A Non-Lethal Procedure for the Determination of Organic Contaminant
Concentrations in Lake Sturgeon (Acipenser fulvescens)
Preface:
The following procedure is designed specifically for the determination of organohalide
contaminant concentrations in Lake Sturgeon (Acipenser fulvescens). This method is not
appropriate for determination of compounds where soaps or plastics might introduce outside
contamination. Because stainless steel will not leave a residue, it is the metal of choice for all
metallic implements. Avoid the use of softer metals such as brass that contain lead and other
substances that are typically the target of contaminant monitoring or those metals that will leave
residues posing a health risk to the fish. In any case, make sure that the sampling tube is
thoroughly washed before use to remove any oil residues applied during manufacture, as these
will typically contain target compounds.
Although it should be considered suitable for all species of sturgeon, the method may not be
suitable for non-Acipenserid species. Suitability is determined primarily by the placement of the
dorsal fin. If the fin is positioned over the body cavity as in lake trout (Salvelinus namaycush),
the species is not a candidate. If there are two dorsal fins as in walleye (Stizostedion vitreum
vitreum) and it can be determined that the second fin is positioned posterior to the body cavity,
the species may be a candidate. The size of the species must also be considered. For obvious
reasons, smaller species must be eliminated. In fact, walleye may be too small. As well,
discretion must be used when working with younger sturgeon. Do not use this technique on very
small juveniles as, again, it will cause too much damage. The method has not been attempted on
fish smaller than 93.1 cm (36.6 in) total length. Using this criterion provides a definitive and
reproducible location for the sample and increases the chance that the fish can be identified later
as having been sampled. Figure 1 illustrates the proper location of the sample.
The method is designed to harvest 4-6 grams of muscle tissue without reducing the fish’s ability
to swim and without the need for primary closure of the resultant wound(s). In some cases the
researcher may find that this mass of tissue is insufficient for analysis. In this case, a second
sample, preferably on the opposite side of the dorsal find, may be taken. More than two samples
per individual are not advised. It should be noted that the researcher is well advised to externally
tag sampled fish, preferably near the anatomic sample site, so that the specimen may be
identified upon recapture. Repeat sampling of the same fish may cause extensive muscle
damage that will hinder its ability to swim and yield duplicate data that is of little scientific use.
The method is properly performed with at least two people and should require no more than one
and one-half minutes from the time the fish is removed from the water until it is returned to the
water.
Water is used for three purposes in this method and it is important to keep the various types
separate. First, municipal water is used to wash all equipment and containers. After this, the
items are rinsed in Reverse Osmosis (preferable), Distilled or de-Ionized water (and other
solvents) to remove residues of soap and ions left by the municipal water. It is important to
check the quality of your rinse water ahead of time, as any contamination from this water will
render the samples useless. None of the aforementioned wash or rinse water should ever come in
Page 43
contact with the sturgeon. The wash water will contain chemicals that pose a health risk to the
fish and the clean rinse water will cause osmotic imbalance. The fish’s native stream water
should be used to dilute the clove oil anesthesia and to irrigate the eyes during the procedure.
This will minimize the chance of poisoning or osmotic shock. If several samples are to be taken,
it is acceptable to use stream water as wash water between samples so long as the implements are
thoroughly solvent rinsed afterward. Carrying a third jug of municipal water is unnecessary
since the stream water should not contain any contaminants not already found in the fish and
those present should be at lower levels than what is already in the fish.
Remember to wear OSHA approved safety glasses at all times during this process.
Material:
 (3) Spray Bottle 22oz or greater Fisher #02-926-4 or equivalent
 100 ml Clove Oil Sigma Aldrich #C-8392
 900 ml 95% Ethanol. Do not use absolute alcohol
 500 ml Acetone Fisher #A929-4 or equivalent
 500 ml 1N Nitric Acid Fisher #A200S4X-212 or equivalent
 1000 ml Reverse Osmosis (RO), Distilled, or de-Ionized water
 Liquinox or other equivalent soap
 (2) Plastic Jugs Fisher #03-327-15 or equivalent
 3/8” – 1/2” (inside diameter) steel tube x at least 12” long, sharpened at one end*
 Wood or plastic cutting board
 Test Tube brush
 Towel or rag
 Steel or plastic bucket, at least one-gallon capacity, for cleaning equipment onsite
 Small glass sample jar with screw-on lid circa 4oz (one per sample)
 Aluminum foil squares large enough to cover mouth of glass jars (roughly 4 square
inches)
 Labels
 Small steel rod and forceps
 Cooler, large enough for all sample jars, and ice
 Drip pan for collection of spent solvents
 Small plastic or steel instrument tray
 Sharpening stone
 Very fine grit sandpaper (400 or greater) or Emery Cloth
 Measuring tape or other suitable device for measuring length
 Scale for measuring weight (optional but highly recommended)
 External tagging device
 Safety glasses
 Scalpel
*Choose a tube as close to 0.5” (12.7 mm) as possible but DO NOT exceed 0.5” or you risk
damaging too much muscle tissue. For the same reason, and for ease of sharpening, choose the
thinnest gauge possible.
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Preparations before leaving the lab:
 Sharpen one end of the steel tube and anneal the sharpened end with a blow torch to help
maintain the edge.
 Be sure that the 22oz (650ml) level is clearly marked on the spray bottle.
 Label one plastic jug to indicate that it is clean water used for rinsing equipment before
use (“RO Water”, “Rinse Water”, etc) and the other “Stream Water”. It is important not
to confuse these two containers.
 Label another container “Waste Solvent”.
 Wash all containers, tools, glass sample jars, and cutting boards with soapy water. Rinse
thoroughly.
 Rinse each item three times each with Nitric Acid, RO or Distilled water, Acetone and
again with RO or Distilled water.
 Rinse small square(s) of Al foil, big enough to cover mouth of the glass jar, with RO or
Distilled water and then with Acetone. Immediately cover the mouth of the glass jar(s)
with the foil and replace the top.
 Fill the Rinse Water jug with Reverse Osmosis (preferred), Distilled or de-Ionized water.
 Fill one spray bottle with approximately 500ml of Acetone and seal tightly.
 Fill another spray bottle with approximately 500ml of 1N Nitric Acid and seal tightly.
 Make a 10% solution of clove oil in Ethanol (ie 1:9 ratio).
 Add 5ml of the 10% clove oil solution to a spray bottle and seal tightly so that the ethanol
does not evaporate.
 Fill a cooler with ice to keep samples cold after collection.
Preparations at the sampling site:
 Record date, water temp, sampling depth, sampling method, gear specifications, name
and contact information of person in charge, and sampling location (GPS if possible, or
other geographic landmark).
 Fill the plastic jug marked “Stream Water” with stream or lake water.
 Fill the spray bottle containing the clove oil solution up to the 22oz (650ml) level with
stream water. This yields a final concentration of 800mg/L clove oil (approximately
0.658% Ethanol). Any higher ethanol concentration will cause damage to and
hemorrhaging of the gills.
 Set the spray bottle to the finest mist possible. Do not use a stream setting under any
circumstances. This precaution will further reduce the risk of gill damage.
 Make sure the end of the tube is as sharp as possible, with no burs.
 Soak a towel with stream water (not RO or Distilled water as this could be harmful to the
fish).
 Wash and solvent rinse the cutting board.
 Wash and solvent rinse the instrument tray. Wash the tube, rod, and forceps in soapy
water and rinse thoroughly. Follow with a solvent rinse and set the instruments in the
tray to keep them clean.**
**Be certain to collect all solvents in a drip pan and pour into the jug marked “Waste Solvents”
for disposal. Check with your lab safety officer for the proper procedure.
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Procedure:
 Record the length and, if possible, the weight of the fish. Metric measurements are
preferred.***
 Cover sturgeon’s eyes with the wet towel.
 Locate the dorsal fin. Place the cutting board underneath sturgeon directly below the
dorsal fin. This will prevent damage to the surface under the fish or to the tube and will
prevent contamination of the sample.
 Carefully open the gill covers. To prevent the Ethanol in the clove oil solution from
damaging the sturgeon’s corneas, begin irrigating the fish’s eyes with stream water from
the jug prior to clove oil application.
 Shake the spray bottle to mix the clove oil solution and IMMEDIATELY spray the gills
2-4 times with the clove oil, depending on size and how much the fish struggles. Because
this concentration will provide only light anesthesia, more may need to be applied during
the procedure. One person must monitor the condition of the gills at all times. If at any
time hemorrhaging from the gills is noted, discontinue clove oil application and irrigate
gills with water (not RO or Distilled water).
 Remove any excess slime and dry the area next to the dorsal fin, on the left side.
 Place the sharp end of the tube on the skin next to the dorsal fin, at its midpoint, and
directly above the cutting board. If open wounds exist or there are lamprey scares at this
location, choose a spot just anterior to the defect. If this is not possible, move to the
opposite side of the dorsal fin.
 Make a small incision with a scalpel, approximately 1.5 cm long, parallel to the midline
that just penetrates the skin.
 Work the tube into this incision and then quickly thrust the tube STRAIGHT down
through the muscle. Do not take an angled approach, as you will harvest less tissue while
causing greater muscle damage. This motion should penetrate through the muscle and
the ventral skin.
 Pull the tube all the way through the fish to prevent the tissue sample backing out of the
tube inside the fish.
 Open the glass jar, remove the foil and, using the forceps, remove the tissue from the tube
and put it in the glass jar. If you can’t reach in far enough to get all of the tissue, drop the
metal rod into the top of the tube to drive the tissue out. Do not allow the tissue to touch
any surfaces that have not been solvent rinsed, as this will introduce contamination.
 Replace the foil and lid.
 Place the sample on ice immediately.
 To allow for proper drainage of the wound and thereby prevent an abscess that might
promote infection, do not attempt primary closure of the wound (suture, glue, tape,
staples, etc).
 Tag the fish externally to allow for later identification of sampled fish. Tag numbers
MUST be retained to prevent future duplication of data.
 Remove the towel and return the sturgeon to the water immediately to clean the gills of
any remaining clove oil. If the fish is not going to be returned to the water immediately,
irrigate the gills. The sturgeon will recover from the anesthesia within one minute.
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
Monitor the fish for one to two minutes (or until it swims away) to assure that it recovers
from the anesthesia and does not bleed excessively from the sample site. Blood loss
should be <10cc.
 If another sample is to be taken, the tube, rod, forceps, and cutting board MUST be
washed and solvent rinsed prior to the next procedure. Be sure to use the test tube brush
to clean the inside of the tube.
 Repeat the procedure as above after making certain that the tube is clean and sharp.
***Length and weight measurements may be taken either before or after the procedure. If
weight is measured, it must be noted whether this value was taken pre or post procedure. Be sure
to record which length value is used (total, fork, dressed, etc.).
Post Procedure:
 Label sample jars and freeze sample as soon as possible. Samples must remain frozen
until analysis.
 Properly dispose of waste solvents and wash water.
Page 47
Dorsal Fin
Entrance
Exit
Steel Tube
Figure 1. Location of sampling site.