[From the U.S. Government Printing Office, www.gpo.gov]

























































                 QL
                 696
                 132
                 M66
                 1992










                                                                         TASK 1. B. 4
                                                                          NA170ZO338-01





         WISCUS61N
              COASrAL
 IMANAGEMENT                                           MONITORING BALD EAGLES ON
                                                         *19CONSIN'S GREAT LAKE@


                                                                  Grant recipient:

                                                       WISCONSIN DEPARTMENT OF
                                                           NATURALRESOURCES





                                                            SEPTEMBER 1992







                                                    This project was funded inpart
                                                    with a grant under the Coastal
                                                    Zone Management Act of 1972,
                                                    as amended, from the U.S.
                                                    Department of Commerce, Office
                                                    of Ocean and Coastal Resource
                                                    Management, Wisconsin Coastal
                                                    Management Program




        WISCONSIN DEPARTMENT
             OFADMINISTRATION
                 P.O. BOX 7868
             MADISON,W1 53707
                  608-266-8234.









                                 Acknowledgement


           This Research Project was funded in part by the Wisconsin
           Coastal Management Program pursuant to Grant #NA170ZO338-01
           from the National Oceanic and Atmospheric Administration.

           The Wisconsin Coastal Management Program, part of the
           Wisconsin Department of Administration, was established in
           1978 to preserve, protect and and develop the resources of
           the Lake Michigan and Lake Superior coastline for this and
           future generations. The Wisconsin Coastal Management
           Program analyzes state policy on Great Lakes issues,
           coordinates government programs that affect the coast, and
           provides grants to stimulate better state and local coastal
           management.
     I/N
















             WISCONSIN BALD EAGLE PRODUCTIVITY AND CONTAMINANT EXPOSURE
                           GREAT LAKES VS. INLAND NEST SITES
                                      1990 - 1992






                                   1992 PINAL REPORT








                                     submitted to:

                         Wisconsin Coastal Management Program
                        Wisconsin Department of Administration
                  Division of Energy and Intergovernmental Relations







                                          by:'

                                Michael W. Meyer Ph.D.
                                  Bureau of Research
                      Wisconsin Department of Natural Resources
                                   1350 Femrite Road
                                  Madison, WI 53716





                                   December 31, 1992











                                     EXECUTIVE SUMMARY

                The International Joint Commission (IJC) Science Advisory
           Board has recommended using the bald eagle as a biosentinel of
           ecosystem health and water quality in the Great Lakes. The
           Wisconsin DNR is currently collaborating with Michigan State
           University (MSU) , University of Wisconsin, University of
           Minnesota, and the National Park Service (NPS) to develop
           methodologies and a protocol for using the bald eagle as a
           biosentinel in the Great Lakes basin. The objective of this
           project is to enhance protocol development by measuring Wisconsin
           Great Lakes bald eagle exposure and relating exposure to recent
           reproductive history, and measures of fish and sediment
           contamination.
                Plasma samples were collected from 97 Wisconsin bald eagle
           nestlings 1990-1992. Under this grant, 60 nestling blood plasma
           samples collected 1990-91 were analyzed for total PCBs
           (polychlorobiphenyls) and DDE (a breakdown metabolite of the
           insecticide DDT) at MSU Pesticide Research Center. All plasma
           samples (100%) contained detectable levels of total PCBs while 44
           samples (73%) had detectable levels of p,pl-DDE [Instrument
           Detection Level (IDL) of total PCB = 5 ug/L, p,p'-DDE = 2.5
            g/L]. Plasma concentrations of total PCBs ranged from 18 - 330
           ug/L while p,pl-DDE concentrations ranged from <IDL - 77 ug/L.-
           U

           The geometric mean nestling plasma total PCB concentration was 66
           ug/L, the geometric mean p,pl-DDE plasma concentration was 6
           ug/L. The geometric mean plasma concentration of total PCBs was
           greatest in bald eagle nestlings sampled along the industrialized
           portions of the Wisconsin River (Rhinelander - Castle Rock
           Flowage) and lower Fox River (both rivers have historic PCB point
           sources; mean total PCB = 125 ug/L, n=11) , followed by nestlings
           sampled along Lake Superior (mean total PCB = 84 ug/L, n=16),
           nestlings sampled at nests >8 km from the shoreline in counties
           adjacent to Lake Superior (mean total PCB = 47 ug/L, n=13) and in
           counties adjacent to Lake Michigan (mean total PCB = 46 ug/L,
           n=13), while the lowest mean concentration was found in nestlings
           sampled along lakes or rivers which have not received point
           source discharges and are remote from agricultural activity
           (total PCB = 33 ug/L, n=7). Between 77-100% of nestlings sampled
           along the Great Lakes shoreline and in adjacent counties had
           detectable levels of plasma p,pl-DDE. This compares to 58% of
           nestlings sampled on the Wisconsin River and lower Fox River,.
           while only 17% sampled at inland sites remote from point sources
           had detectable levels of plasma p,pl-DDE. Nestlings sampled
           along the Lake Superior shoreline had the greatest geometric mean
           plasma concentration of p,pl-DDE (mean p,pl-DDE = 16 ug/L, n=16),
           followed by nestlings sampled at nests >8 km from the shoreline
           in counties adjacent to Lakes Superior (mean p,pl-ODE = 6 ug/L,
           n=13) and in counties adjacent to Lake Michigan (mean p,pl-DDE
           7 ug/L, n=13). Even lesser concentrations were found in
           nestlings sampled in inland counties (below point source
           discharge, mean p,pl-DDE     5 ug/L, n=11; no point source, mean
           p,p'-DDE <2.5 ug/L, n=7).











                                                                               2

                There was no relationship between plasma total PCB and the
           reproductive history (# young hatched/nest attempt, 1985-1992) of
           the nest territory it was collected in    (r2 = 0.07, n=48), however
           the relationship was somewhat greater between plasma p,pl-DDE and
           nest territory reproductive history (r2=    0.22, n=48).
                Bald eagles nesting in the region of Wisconsin which had the
           greatest mean plasma PCB concentration (the Wisconsin River/Fox
           River) have a healthy reproductive rate (1985-92 productivity
           1.36 + 0.16 young hatched/active territory, 1992 = 21
           territories) however many territories have existed <10 years and
           data indicates that reproductive performance of PCB-exposed
           eagles declines over time. Because loss of successfully breeding
           adults has a greater impact on bald eagle population dynamics
           than does nestling mortality, scientists at Michigan State
           University are developing an'index of adult survival, a DNA
           fingerprinting technique which will allow for determination of
           adult turnover rates at nest sites. This data is required to
           accurately assess the impact of contaminants at a given nest
           territory.
                Nestlings in the Lake Superior shoreline region had the
           greatest mean plasma p,p'-DDE concentrations and a lesser rate of
           productivity (0.94 + 0.18 young/nest attempt, n=8 years) however
           prey availability appears to be limited in that region. Lake
           superior nestlings received 72% fewer prey deliveries than did
           inland Wisconsin nestlings in 1992; the average prey delivery-
           rate for the Lake Superior shoreline and Apostle Islands nests
           was 1.65 prey items/16 hours of observation while 5.90 prey
           deliveries/16 hours occurred inland. of the identified prey
           items delivered to Lake Superior bald eagle nests, 80% were fish,
           14% were birds, and 6% were mammals. This compares with inland
           prey deliveries which were 97% fish, 2% reptiles (turtles), 0.8%
           mammals, and 0.2% birds. The size class of the Lake Superior
           prey items include: 0-611 = 17%, 6-1211 = 68%, 12-1811 =.12%, and
           >1811 = 3%. The size class of inland prey items include 0-611 =
           4%, 6-1211 = 61%, 12-1811 = 33%, and >1811 = 2%.   Therefore, Lake
           Superior nestlings received not only 72% fewer prey deliveries
           but also smaller prey items (85% were <1211 on Lake Superior, 65%
           were <1211 inland). While fish dominated the diets of eaglets at
           both inland and'Lake Superior nests, birds were a more common
           diet component on the Lake Superior shoreline (14%) than inland
           (0.2%).
                Two nestling plasma samples collected from nests along the
           shoreline of Green Bay are currently undergoing chemical analysis
           at Michigan State University. This region has the poorest bald
           eagle reproductive history in Wisconsin and fish from the region
           have the greatest concentrations of PCBs and DDE in the state
           today.












                                                                           3




                                  PROJECT OBJECTIVES


               Recently, the International Joint Commission Science
          Advisory Board has recommended using the bald eagle as an
          bioindicator of ecosystem health and water quality in the Great
          Lakes. The Wisconsin Department of Natural Resources (DNR) is
          among several regulatory agencies charged with establishing water
          quality criteria protective of wildlife in the Great Lakes. The
          Wisconsin DNR is currently collaborating with Michigan State
          University (MSU), University of Wisconsin, University of
          Minnesota, and the National Park Service to develop methodologies
          and a protocol for using the bald eagle as a biosentinel of Great
          Lakes ecosystem health. The primary objective of this project is
          to enhance protocol development by measuring Wisconsin Great
          Lakes bald eagle organochlorine (OC) contaminant exposure and
          relating exposure to recent reproductive history, and measures of
          @ish and sediment contamination. Specific goals include
          increasing the number of Wisconsin nestling bald eagle plasma
          samples collected to 100 for 1990-1992 and to conduct chemical
          analysis of 60 plasma samples. The samples were analyzed for
          total PCBs (polychlorobiphenyls) and p,pl-DDE (a breakdown
          metabolite of the insecticide DDT,
          dichlorodiphenyltrichloroethane) at MSU Pesticide Research
          Center. These contaminants have been implicated in reducing bald
          eagle reproductive performance in the Great Lakes basin and are
          known to cause eggshell thinning, embryo mortality, and
          teratogenicity. Results of this analysis will be used to provide
          baseline data regarding Wisconsin Great Lakes bald eagle nestling
          OC exposure that exposure will be compared to'that of nestlings
          further inland. In addition, the Wisconsin bald eagle nestling
          plasma total PCB and p.,pl-DDE concentrations were related.to the
          reproductive history (1985-92) of the nest territories at which
          the samples were taken. Finally, eaglet plasma contaminant
          concentrations were compared to the contaminant levels of game
          fish and sediment samples taken in the region of the nest sites.
          Lake Superior bald eagle foraging behavior and nesting ecology
          were also documented and 1992 Wisconsin Great Lakes bald eagle
          productivity was measured.



                                      BACKGROUND


               Industrial and agricultural pollution has contaminated the
          sediment and water column of several Wisconsin Great Lakes
          harbors and tributaries. Despite the ban on industrial uses of
          PCBs and agricultural use of the pesticide DDT in the 1970s,
          these substances persist in the Great Lakes foodweb and pose a
          health risk to humans consuming some Great Lakes fish. In
          response, the Wisconsin DNR has issued fish consumption
          advisories for sport fisherman along portions of Wisconsin's
          Great Lakes shoreline. Similar advisories have been issued for












                                                                           4

          portions of the Menominee, Fox, Sheboygan, and St. Louis Rivers
          which discharge into the Great Lakes. While these compounds pose
          a health risk to humans, it is also suspected that they
          negatively impact the health and productivity of top-predator
          wildlife which are dependent on the Great Lakes foodweb.
               Bald eagles (Halieeatus leucocephalus) nesting in the Great
          Lakes coastal region (<8 km from the shoreline) are less
          productive and experience greater contaminant exposure than do
          bald eagles which nest further inland (International Joint
          Commission, 1989). Consumption of contaminated prey from the
          Great Lakes (primarily fish) is the likely route of contaminant
          exposure.. Kozie and Anderson (1991) found that Wisconsin Lake
          Superior bald eagle productivity was reduced, and nestling
          mortality and contaminant exposure was greater when compared to
          Wisconsin bald eagles nesting inland. They concluded that
          consumption of contaminated prey (primarily herring gulls) was
          the likely cause of reduced Lake Superior productivity. A more
          recent analysis of Wisconsin bald eagle productivity (1985-1992)
          showed that 20 active bald eagle nests were located within 8 km
          of the Wisconsin Great Lakes shoreline (8 kin is thought to be the
          feeding range of nesting bald eagles). During that time, eagles
          nesting on Wisconsin's Lake Superior shoreline produced 0.94 ,
          young/nest attempt (n=110 nest attempts) while eagles nesting on
          the Lake Michigan shoreline produced 0.27 young/nest attempt
          (n=11 nest attempts). In contrast, eagles nesting in the North
          Central District of Wisconsin (all nests are >40 km from the
          Great Lakes) produced 1.32 young/nest attempt (n=1011 nest
          attempts). Productivity of 0.7 young/active nest is required for
          population stability (Sprunt et al. 1973) and 1.0 young/active
          nest is considered typical productivity for a "healthy" eagle
          population.
               Unhatched bald eagle eggs collected from Wisconsin. Great
          Lakes eagle nests 1970-1988 had total PCB, p,pl-DDE, and dieldrin
          concentrations 6 - 50x greater than eggs collected inland
         '
          (Wiemeyer et al. 1984; WDNR unpubl. data). Blood plasma
          collected from 5 Wisconsin bald eagle nestlings in 1989 contained
          PCB and DDE levels 5x greater than plasma collected from nestling
          eagles at inland sites (Bowerman et al. 1989). Bald eagle
          nestling blood plasma provides a "top of the food chain" measure
          of contaminant exposure specific to a given site because nestling
          plasma contaminant levels likely reflect that of prey collected
          within the adult feeding territory (generally within 8 km of the
          nest).



                                      METHODOLOGY


                               Blood Plasma Collection

               A total of 97 nestling plasma samples were collected at
          Wisconsin bald eagle nest sites 1990-92 (Figures 1,2,& 3).
          Contracted climbers or WDNR personnel (D. Evans, Hawk Ridge











          Observatory, Duluth, MN; R. Eckstein, WDNR) captured bald eagle
          nestlings by climbing to the nest and restraining the nestlings
          when they were 4-7 weeks old. The climber placed a US Fish and
          Wildlife Service band on the nestling and lowered it to a ground
          crew in a nylon bag. On the ground, two field technicians
          weighed the eaglet and drew ten cc's of whole blood from the
          bracialus vein. One technician held the eaglet and secured the
          talons while the other drew blood. Ten cc's of blood were
          obtained with sterile 10 cc syringes (glass or plastic) tipped
          with 20 ga needles. The whole blood was placed in a 10 cc
          heparinated vacutainer for organochlorine pesticide and total PCB
          analysis. All blood containers were then labelled with the
          bird's band number, nest location, state, date, and initials of
          the technicians present. Three contour breast feathers were
          collected from the nestling for mercury analysis. Six
          measurements were obtained from each nestling to determine their
          age and sex; hallux claw arc, foot pad length, eighth primary
          feather length, sixth tail feather length, culmen depth, and
          length of the gape. The nestling was then returned to the nest.
               Blood samples were placed on ice and refrigerated upon
          return from the field. Within 48 hours the whole blood was
          centrifuged for 10 minutes at 1200 rpm and plasma was transferred
          to another green top vacutainer with a clean pipette. This
          vacutainer was labeled as above and frozen upright. The red
          blood cells were removed from the vacutainer and placed in a
          centrifuge tube. The empty green top vacutainer was rinsed with
          normal saline solution into the centrifuge tube. The centrifuge
          tube was then spun for 10 minutes. The saline layer was decanted
          off using a clean pipette and 5 ml of saline solution was added
          to the tube. The tube was centrifuged for an additional 5
          minutes and the saline layer decanted off again. At this point
          0.1 ml of buffer was added and the centrifuge tube was labeled
          and frozen upright. At the end of the field season all frozen
          samples were shipped on dry ice via Federal Express to Michigan
          State University for contaminant analysis.
               Samples were collected in different regions to compare bald
          eagle nestling exposure in various Wisconsin-habitats. First,
          nestling,plasma was 'obtained from all productive eagle nests
          a.long Wisconsin's Lake Superior and Lake Michigan shorelines.
          Second, nestling plasma was collected from a sample of eagle
          nests in counties adjacent to  'the Great Lakes (Douglas, Bayfield,
          Ashland, Iron, Marinette, Oconto Co.) but > 8 km inland (these
          eagles are unlikely to feed directly on the Great Lakes).
          Finally, nestling plasma was collected in inland counties (Vilas,
          Oneida, Lincoln, Marathon, Portage, Wood, Adams, Juneau, Sawyer,
          Rusk, Shawano, outagamie) at two types of sites. one type of
          habitat included riverine nests along the Wisconsin and Fox
          Rivers that are near point source discharges from industry and
          municipalities. The second type of inland site is nests which
          occur on waterways which receive no point source discharge within
          40 km of the nests.











                                                                            6

                                Blood Plasma Analysis

                Concentrations of organochlorines were determined by gas
           chromatography with confirmation of pooled samples by mass
           spectrometry at MSU Pesticide Research Center (Price et al. 1986;
           MDPHL 1987). Individual 2-4 ml plasma samples were dissolved in
           methanol and extracted twice with 5 ml of a 1:1 mixture of
           hexane-ethyl ether by agitating on a rotary mixer for 20 minutes
           at 50-55 rpm. Extracts were concentrated on a hot water bath to
           a volume of 0.5 ml. Clean-up was done on a 7 mm Chromaflex
           column packed with 2.5 g of Florisil using 10 ml of hexane.
           Elution of polycholorinated biphenyls (PCBs) and chlorinated
           hydrocarbon pesticides from the Chromaflex column was
           accomplished with 20 ml of 6% ethyl ether/hexane. Elution of
           dieldrin from the column was accomplished with 20 ml of 20%
           ethyl ether/hexane. Separation of PCB from the chlorinated.
           hydrocarbon pesticides was accomplished with a Chromaflex column
           packed with Silica Gel 60. Elution of hexachlorobenzene (HCB)
           and mirex was accomplished with 15 ml hexane. Elution of Aroclor
           1260, Aroclor 1016, and polybrominated biphenyl (PBB) was
           accomplished with an additional extraction with 20 ml hexane.
           Elution of Aroclor 1016 and chlorinated hydrocarbon pesticides
           was accomplished with 20 ml of benzene (MDPHL 1987). Gas
           chromatography was performed on a Varian 3700 gas chromatograph
           equipped with small volume pulsed 63 Ni electron capture detectors,
           Varian 8000 Auto Sampler, and CDS-111 microprocessor. A 1.83 m x
           0.64 cm x 2 mm i.d. glass column packed with 3% SE-30 was used.
           Nitrogen flow rate was 30 ml/min through the column during ,
           operation. Total PCB concentration was determined on the basis
           of mean weight percent factors (Webb and McCall 1983) and other
           individual peaks were determined by reference to the relative
           retention time of p,pl-DDE x 100 (MDPHL 1987).   We measured
           concentrations of total PCBs (measured as Aroclor 1254 or 1260),
           1,11-(2;2,2-Trichloroethylidene)bis(4-chlorobenzene] (p,pl-DDT),
           and its metabolites p,pl-DDD and p,pl-DDE, HCB, heptachlor
           epoxide, cis-nonachlor, trans-nonachlor, oxychlordane, dieldrin,
           PBB, toxaphene, mirex, alpha-chlordane, and gamma-chlordane.
           (See Appendix for Standard Operating Procedure and certification
           of results by MSU QA/QC officer).

                           Food Habits and Foraging Behavior

                Food habits of nesting bald eagles have often been
           determined by collecting prey remains in the nest and on the
           ground around the nest tree and quantifying the prey items
           present. In a recent study (Mersmann et al. 1992) it was
           concluded that this technique is biased towards over-
           representation of birds, medium sized mammals, and large bony
           fish while small mammals and small fish were under-represented.
           Because of the bias, two additional techniques were used to
           determine Wisconsin Great Lakes bald eagle food habits.












                                                                           7

          Technicians conducted dawn-to-dusk observations at several Lake
          Superior shoreline nests to document bald eagle food habits while
          video cameras mounted at nest sites were used to document eagle
          feeding behavior at inland.1nest sites.

          Direct Observation Protocol

               Observations of bald eagle nesting and feeding behavior
          were conducted at seven nests on the Wisconsin Lake Superior
          shoreline, including two on the Apostle Islands National
          Lakeshore (APIS), during the 1992 nesting season. Observations
          were conducted at all known nest sites within 8km of the shore of
          Lake Superior where suitable blind sites were available.
          Location of the nests is shown in Figure 3 and include DU-9b
          (T45N R10W S18), BY-23 (T49N R09W S05), BY-21a (T51N R7W S35),
          BY-3b (T52N R4W S16), BY-15b (T47N R5W S08), AS-20c (T51N R1W
          S14), and IR-32 (T47N R1.E S12).
               Direct observations were made with binoculars and spotting
          scopes. scopes included a Nikon Field Scope, a Kowa TSN-1, a
          Bushnell Spacemaster, a Celestron SS-80, and a Meade 97E. All
          scopes had magnifications ranging from 20x-60x with 40x the most
          common selection for viewing the nests from the distances of the
          blinds. Incubation observations were conducted at four nests in
          April. After hatching, 24 hour observation periods began; 1300
          dusk on day 1, dusk - dawn on day 2. Each observation shift was
          approximately four hours long for a single observer, observers
          then switched. Training of observers was conducted by an
          experienced observer familiar with the procedure. Every five
          minutes the observer would enter a code descriptive of the major
          activity of each eagle on the territory during the previous five
          minutes. Food habits were closely monitored; time of prey
          delivery, prey type, and size class were recorded.

          Video Camera Protocol

               Six video cameras were mounted February 1-15, 1992, at bald
          eagle nest sites in northern Wisconsin (Figure 3) before the
          initiation of nesting. Two-cameras were mounted in nest trees
          and four were mounted in trees adjacent to the nest tree. Three
          territories were on the Turtle Flambeau Flowage, Iron County (IR-
          9c, IR-33, IR-35a), one was on a lake in Vilas County (VI-61c),
          and two were on lakes in Oneida Co (ON-49, ON-79). All six
          territories produced chicks in 1992. Cameras were camouflaged
          with natural materials to minimize their visual impact. Spruce
          and balsam fir bows were attached to 111 chicken wire and wrapped
          around the Sony M-350 black and white video cameras, pine boughs
          and cedar bark were used to camoulflage the smaller M-332 Sony
          cameras. Coaxial cable (type RG-8UM, Tandy Corp.) connected the
          video cameras to Sony 8mm timelapse recorders modified for field
          use by Fuhrman Diversified, Laporte, TX. Recorders were placed
          200-300m from the nest tree to minimize disturbance to the nest
          site when batteries and tape were changed. The cable was












                                                                             8

           anchored to the trunk of the tree with it' electric staples as
           cable run on the ground was damaged by white-tailed deer and
           snowshoe hare in the spring. An 8mm. pseudo-time lapse video
           recorder (model WCMS-4/V11, FUhrman Diversified, Inc.) was loaned
           to the research project from the National Park Service and 2
           Fieldcam TLV 8mm timelapse video recorders (Fuhrman Diversified,
           Laporte, TX) were purchased by the WDNR with grant support from
           the Great Lakes Protection Fund. The recorders were set to
           record 1 frame/second and tape and batteries were changed every
           3-4 days. Fuhrman Diversified manufactured customized
           weatherproof housing for the Sony recorders and modified the
           recorders so that they could be powered with 12 V DC batteries.
           When operated in the field the recorders and batteries were
           placed in custom security boxes constructed of 16 gauge steel
           sheet metal which were hinged, padlocked, and chained to large
           trees. An internal time/date stamp superimposed the date and
           time over the video image. Video recordings were then analyzed
           using a video monitor and bald eagle time/activity budgets, food
           habits (time and number of daily prey deliveries, prey size, prey
           type), and behavior were quantified and compared to data gathered
           along the Lake Superior shoreline.

           Nest Prey Remains

                Prey remains.were collected at all nest sites that nestling
           plasma was obtained. Bones, scales, and feathers will be
           identified to provide an estimate of the type of prey delivered
           @o the chicks at the nest site. -This information will provide an
           index with which to assess the primary routes of contaminant
           exposure at a particular nest site. Analysis of material is
           underway at Michigan State University.

                          Great Lakes vs. Inland Productivity

                Two aerial surveys, one conducted in mid-April and one in
           late-May, were used to obtain detailed information on the
           Wisconsin Lake Superior and Lake Michigan bald eagle population
           status (number of occupied territories) and reproductive
           performance (% nest success, number of young produced/occupied
           bald eagle territory). An additional flight was conducted on May
           8 to identify nests which had been abandoned without producing
           young. A climber entered these nests and searched for unhatched
           eggs for contaminant analysis. In addition, all nests were
           revisited when chicks were 4-6 weeks of age to collect nestling
           plasma samples. In order to continue yearly comparisons of
           inland and Wisconsin Great Lakes bald eagle productivity, the
           WDNR conducted two aerial surveys of all bald eagle terr 'itories
           in Wisconsin's North Central District, approximately 150
           territories representing 30-40% of the state's total breeding
           population.











                                                                           9

                          Pish and Sediment Contaminant Data

               All available data was gathered on the concentrations of
           contaminants in fish and sediment in the general regions that
           bald eagle nestling plasma was collected. Primary data sources
           included the Wisconsin DNR Fish Contaminant Database, the Green
           Bay Mass Balance Proj ect, and the WDNR Bureau of Water Resource
           Management.



                                 RESULTS & DISCUSSION

           1990 - 1991 Wisconsin Nestling Plasma Contaminant Concentrations

               sixty Wisconsin bald eagle nestling plasma samples collected
           1990-91 were analyzed for total PCB and p,p'-DDE content at
           Michigan State University Pesticide Research Center. All plasma
           samples (100%) contained detectable levels of total PCBs while 44
           samples (73%) had detectable levels of p,pl-DDE (Table 1;
           Instrument Detection Level (IDL) of total PCB = 5 ug/L, p,pl-DDE
            2.5 ug/L]. Plasma concentrations   of total PCBs ranged from 18
            330 ug/L while p,pl-DDE concentrations ranged from <2.5 (IDL)
           77 ug/L. The geometric mean nestling plasma total PCB
           concentration was 66 ug/L, while the mean p,pl-DDE plasma
           concentration was 6 ug/L.
               Samples were categorized according to region of collection
           (as described in methods section) and the frequency of detection
           and geometric mean plasma contaminant concentrations were
           compared. As stated previously, PCBs were detected in the plasma
           of 100% of nestlings sampled in all regions of Wisconsin. The
           greatest geometric mean plasma concentration of PCBs was found in
           bald eagle nestlings sampled along rivers with historic point
           sources of PCBs (the industrialized portion of the Wisconsin
           River (Rhinelander - Castle Rock Flowage)and the lower Fox
           River](mean total PCB = 125 ug/L, n=11, Table 2, Figure 4),
           followed by nestlings sampled along Lake Superior (mean total PCB
           = 84 ug/L, n=16), nestlings sampled at nests >8 km from the
           shoreline in counties adjacent to Lake Superior (mean total PCB
           47 ug/L, n=13) and in counties adjacent to Lake Michigan (mean
           total PCB = 46 ug/L, n=13), and the lowest mean concentration was
           found in nestlings sampled along lakes or rivers which have not
           received point source discharge and are remote from agricultural
           activity (total PCB = 33 ug/L, n=7). Between 77-100% of
           nestlings sampled along the Great Lakes shoreline and in adjacent
           counties but >8 km inland had detectable levels of p,p'-DDE in
           their plasma. This compares to 58% of nestlings sampled at
           inland sites subject to historic point source discharge while
           only 17% sampled at inland sites remote from point source had
           detectable levels of p,pl-DDE (Table 2, Figure 5). Nestlings
           sampled along the Lake Superior shoreline had the greatest
           geometric mean concentration of p,pl-DDE in their plasma (mean
           p,p I-DDE = 16 ug/L, n=16), followed by nestlings sampled at nests












                                                                           10

          >8 km from the shoreline in counties adjacent to Lakes Superior
          (mean p,pl-DDE = 6 ug/L, n=13) and in counties adjacent to Lake
          Michigan (mean p,pl-DDE = 7 ug/L, n=13). Even lesser
          concentrations were found in nestlings sampled in inland counties
          (below point source discharge, mean p,pl-DDE =.5 ug/L, n=11; no
          point source, mean p,pl-DDE <IDL), n=7).
               While nestlings sampled along Lake Superior had mean total
          PCB concentrations nearly 2.6% greater than nestlings sampled at
          remote inland Wisconsin sites, the mean concentration was less
          than half the mean found for other Great Lakes bald eagle
          nestling plasma samples (Bowerman et al. 1990, Table 3), and less
          than concentrations of PCBs in plasma collected along the
          industrialized rivers of Wisconsin, and along the lower Columbia
          River in Oregon/Washington (Table 4). The relatively small
          concentration of PCBs in Lake Superior nestling plasma is
          compatible with fish contaminant data, only large lake trout have
          detectable levels of PCBs along Wisconsin's Lake Superior
          shoreline. Piscivorous avian species, such as berring gulls, do
          have elevated levels of PCBs in Lake Superior, are taken by
          eagles, and may account for PCB concentrations of 100-201 ug/L
          found in 7 of 16 (44%) Wisconsin Lake Superior eaglet plasma
          samples; 4 of the 7 samples were collected on the Apostle Islands
          archipelago. Eagles were observed delivering birds to nestlings
          at all Lake Superior nests we observed in 1992. In comparison,
          total PCB concentrations exceeded 100 ug/L twice as often (91%)
          in samples collected on the industrialized portions of the
          Wisconsin and lower Fox River where many fish, particularly carp,
          have detectable concentrations of PCBs. Some Lake Superior
          eaglets appear to consume little contaminated prey as 25% had
          plasma concentrations within the range (24-66 ug/L) found in
          @estlings in remote Wisconsin nest locations. In addition, there
          is evidence of great year to year variability in exposure on Lake
          Superior, indicating that contaminant exposure may be sporadic.
          While the 1990 nestling plasma PCB concentration at AS-12 (41
          ug/L, N. Twin Island, APIS) was among the lowest in the state, a
          nestling sampled in the same nest in 1991 had total PCB levels 5x
          greater (201 ug/L). It is possible that it had fed solely upon
          fish in 1990 but had fed upon gulls (or other contaminated birds)
          in 1991. The mean total PCB content of nestling plasma samples
          collected along the Wisconsin River and lower Fox River was
          nearly identical to that found along the lower Columbia River in
          Oregon/Washington; these rivers all receive effluent from pulp
          and paper mills which often contained PCBs in the 1960s and 1970s
          (primarily due to de-inking processes). The greatest fish and
          sediment PCB levels found today in Wisconsin occur in portions of
          the lower Fox River and in lower Green Bay. Two nestlings were
          sampled in a nest on the lower Fox River (in Kaukauna) and were
          found to have elevated plasma PCB levels (115 & 124 ug/L). Even
          greater concentrations were found in eaglets at inland sites in
          Marinette Co. (213 ug/L lower Menominee River location, 330 ug/L
          Lake Noquebay) and from eagles on the Wisconsin River (187, 197,
          & 277 ug/L from eaglets on the Petenwell/Castle Rock Flowages;










           146 & 157 ug/L in nestlings from Lincoln and Marathon Co.). The
           nests,in Marinette County were both within 12 miles of the Green
           Bay shoreline and it is conceivable that the nestlings were fed
           fish, or avian species which feed on fish, from Green Bay. The
           eagle nest on the lower Fox River lies midway between the two
           major sediment sources of PCBs on the river and it is possible
           that the locks and dams compartmentalize the sediment thus
           protecting the eaglets from receiving the greatest dose of PCBs
           possible.on that river system. A plasma sample was collected
           from a nestling in a-nest on the mouth of the Oconto River near
           the shore of Green Bay (Lake Michigan) in 1990. The sample was
           placed in the wrong batch at the analytical laboratory and will
           be analyzed in January 1993 as will an additional Green Bay
           shoreline plasma sample collected in 1992 in Marinette Co. It is
           likely that both samples will have greatly elevated total PCB
           concentrations as most Green Bay fish still contain >1 ppm total
           PCB.
                Nearly all nestlings (86%) sampled in counties adjacent to
           the Great Lakes had detectable p,pl-DDE in their plasma, however
           the metabolite was detected in only 44% of samples collected in
           inland Wisconsin counties. The greatest concentrations of P,pl-
           DDE were found in Lake Superior nestling plasma samples; only'9
           Wisconsin samples exceeded 20 ug/L, however 7 of 9 samples (78%)
           were collected along Lake Superior. This finding is consistent
           with the likelihood that most OC pesticide residues have been'
           metabolized and/or transported from the inland rivers, however a
           slower rate of metabolism and longer residence time is suspected
           for.DDT and DDE in the Great Lakes, particularly Lake Superior.
           In addition, some regions near Wisconsin's Great Lakes shoreline
           were placed into fruit orchards (particularly Door and Bayfield
           Co.) and received heavy OC pesticide application before the
           compounds were banned in the 1970s, some of which persists in the
           offshore and tributary sediment. The mean p,pl-DDE
           concentrations for all regions in Wisconsin, including Lake
           superior, were much less than that found for other Great Lakes
           nestlings (Table 3) and those on the lower Columbia river
           (Table 4). Despite the fact that they had comparable levels of
           PCBs, the mean p,p-DDE concentration of plasma samples collected
           on the Wisconsin/Fox River (4 ug/L) was 96% less than that found
           in lower Columbia River plasma samples (98 ug/L). Samples
           collected from remote Wisconsin sites also contained lesser
           concentrations than those collected in the Michigan and Oregon
           interior.
                There was no relationship between plasma total PCB and the
           reproductive history (#young hatched/nest attempt, 1985-1992) of
           the nest territory it was collected in (r    2 = 0.07, n=48; Figure
           6), however the relationship was somewhat greater between plasma
           p,pl-DDE and nest territory reproductive history (r     2 =  0.22,
           n=48; Figure 7). Bald eagles nesting in the region of Wisconsin
           with the greatest mean plasma PCB concentration (the Wisconsin
           River/Fox River) have had excellent productivity (1985-92











                                                                          12

          productivity    1.36 + 0.16 young hatched/active territory, 1992
          21 active territorie's) however many have been occupied <10 years
          and data indicates that reproductive performance of PCB-exposed
          eagles declines over time. Because loss of successfully breeding
          adults has a greater impact on bald eagle population dynamics
          than does nestling mortality, Michigan State University is
          developing an index of adult survival, a DNA fingerprinting
          technique which will allow for determination of adult turnover
         -rates at nest sites. This data is required to accurately assess
          the impact of contaminants on a given nest site. The region
          where nestlings had the greatest mean plasma p,p'-DDE
          concentrations (Lake Superior nestlings) had poorer productivity
          (1985-92 productivity = 0.94 + 0.18 young/nest attempt, 1991 = 18
          active territories) however prey availability also is limited in
          that region (see Foraging and Food Habits. Two plasma samples
          collected from Green Bay are currently undergoing analysis; this
          region has the poorest.bald eagle reproductive history in
          Wisconsin and fish from the region have the greatest
          concentrations of PCBs and DDE in the state today.

                           1992 Nestling Plasma Collection

               In 1992 we collected blood and feather samples from 33 bald
          eagle nestlings, including all 11 productive nests within 8 km of
          the Great Lakes (Table 5). The full 10 ml of blood was collected
          from all but one nestling from which 9 ml were collected. Three
          contour breast feathers were collected from 30 nestlings, two
          feathers from two nestlings and one nestling had not yet ,
          developed contour feathers so none could be collected. Sample
          analysis is currently being conducted at the laboratory of Dr.
          John Giesy, Michigan State University Pesticide Research Center
          with grant support from the Great Lakes Protection Fund.

                               Foraging and Food Habits

          Direct Observations

               Direct observations were conducted at seven bald eagle nests
          on the Wisconsin Lake Superior shoreline and the Apostle Island
          National Lakeshore (APIS) in 1992. Direct observations were
          initiated 8 April with an incubation observation at IR-32a. A
          total of 72 hours of incubation observations and 872 hours of
          post-hatching direct observations were.made on the Wisconsin Lake
          Superior shoreline. Observers witnessed 92 prey deliveries.
          Eggs failed to hatch at one nest while two nests failed after
          hatching (see 1992 Great Lakes Productivity.below).

          Vi deo Recordings

               Eighty one hours of incubation activity was recorded at
          three territories from late March - mid April. Nearly 2800 hours
          of nest activity was recorded during the post-hatching stage. A











                                                                              13

           total of 601, 627, and 766 hours of nest activity was recorded at
           the IR-9a, IR-33, and ON-79 territories, respectively. These
           sites were monitored continuously through the nesting season.
           Recording began at ON-49 on 2 June because a camera adjustment
           was required (the eagles had added 50 cm of nest material to the
           nest, placing the camera out of position). A total of 308'hours
           of nest behavior was recorded at ON-49. Final camera adjustments
           were also made at the IR-9a and IR-33 nests the last week in May
           as portions of those nests were also not visible. The camera at
           VI-61c was moved to the new nest 2 June and recording began at
           that time; 467 total hours were collected at VI-61c. A camera
           was also mounted at IR-35c, 15 June. This camera was only
           operated for two weeks as only 70% of the nest was visible and
           110 hours of nest activity was recorded.

           Ouantity of Prey Delivered

                The prey delivery rate was calculated for each Lake Superior
           nest by totalling the number of prey deliveries, dividing by the
           total number of hours observed at that nest and multiplying by 16
           to normalize the rate to a 16 hour daylight period. Prey
           delivery rates were 1.62, 1.40, 2.06, 1.75, and 1.36 prey items
           per 16 hours of observation at nests BY-23 (2 yng), AS-20c (2
           yng), BY-15a (2 yng, 1 died), IR-32a (2 yng) and BY-21 (2 yng,
           both died), respectively. The average prey delivery rate for-the
           Lake Superior shoreline and Apostle Islands nests was 1.65 prey
           items per 16 hours of observation (Figure 9).
                The prey delivery rate was also calculated for inland nest
           sites by totalling the number of prey   deliveries recorded on
           video tape, dividing by the total number of daylight hours video
           recordings occurred, and multiplying by 16 to normalize the rate
           to a 16 hour daylight period. Prey delivery rates were 6.87,
           5.43, 7.03, 7.53, and 1.86 prey items per 16 hours of video
           recordings at nests ON-79 (2 yng), ON-49 (1 yng), IR-33 (3 yng),
           VI-61c (3 yng), and IR-9a (2 yng, 1 died) respectively. One
           nestling died at IR-9a (lowest inland prey delivery rate) while
           all nestlings survived at the other nest sites. The average prey
           delivery rate for the inland nest sites was 5.91 prey
           deliveries/16 hours of video recording (Figure 9).

           TVpe and Size of Prey Delivered

                Ninety two prey deliveries were documented at five
           successful,Lake Superior bald eagle nests in 1992. Observers
           were able to identify 64 of the prey items delivered (70%) to
           class (fish, mammal, bird, reptile; Figure 10) and were able to
           determine the size class (0-611, 6-1211, 12-1811, >18"; Figure 11)
           of 66 prey items (74%).    The video cameras mounted at inland
           Wisconsin bald eagle nests recorded 694 prey deliveries at five
           successful nests in 1992. Of the known prey items delivered to
           Lake Superior bald eagle nests, 80% were fish, 14% were birds,
           and 6% were mammals. This compares with inland prey deliveries











                                                                              14

           which were 97% fish, 2% reptiles (turtles), 0.8% mammals, and
           0.2% birds. The size of the Lake Superior prey items include 17%
           in the 0-611 size class, 68% were 6-1211, 12% were 12-1811, and 3%
           were,>18". The size class of inland prey items include 4%
           between 0-6111 61% were 6-1211, 33% were 12-1811, and 2% were >18".
           Therefore, Lake Superior nestlings received not only 7.3% fewer
           prey deliveries but also smaller prey items (85% were <1211 on
           Lake Superior, 65% were <1211 inland). While fish dominated the
           diets of,eaglets at both inland and Lake Superior nests, birds
           were a more common diet component on the Lake Superior shoreline
           (14%) than inland (0.2%).

           Nest Prey Remains

                Nest prey remains were collected by the climber while the
           nestlings were being bled.   The ground crew also collected all
           prey remains visible at the  base of the nest tree. Samples are
           currently being analyzed at  Michigan State University.

                   1992 Wisconsin Great Lakes Bald Eagle Productivity

                Nest success on the Wisconsin Lake Superior shoreline (nests
           within 8 km of the shoreline) in 1992 was 67%; 8 of 12 nest
           attempts produced young. Thirteen bald eagles were fledged from
           the 12 active territories for a productivity of 1.08 young/active
           territory (Figure 12) and 1.62 young/successful nest. This
           compares to 11 successful nests out of 18 active,territories
           (61%) in 1991, with 17 young produced (0.94 young/active nest,
           1.54 young/successful nest). In contrast,,both previously active
           nests on Wisconsin's Lake Michigan (Green Bay) shoreline did not
           produce young in 1992. A climber entered both nests and found
           the one near the mouth of the Oconto River (OC-4) had been worked
           on but there were no indications that eggs were laid (D. Evans,
           pers. comm.). A raptor biologist who lives nearby reported that
           both adults were seen at the nest in early spring (T. Erdman,
           pers. comm.). The other territory is near the mouth of the
           Peshtigo River (MT-7 and 7a) and was not worked on in 1992 (D.
           Evans, pers. co mm.) however one adult was observed on the
           territory in the spring (T. Erdman, pers. comm.). Two new nests
           were located on Green Bay in 1992. One nest in Marinette County
           produced 1 young (plasma was sampled) and one unhatched egg was
           @ollected. The other nest was constructed on Little Tail Point
           in Brown County, no eggs were laid, however 1 adult and 1
           subadult were in residence (T. Erdman, pers. comm.)
                Inland Wisconsin productivity (Northcentral District) in
           1992 remained good, lj9 out of 176 active territories produced a
           total of 233 young; nest success = 79%, productivity = 1.32
           young/active territory (Figure 12), 1.68 young/successful
           territory. In 1991, inland Wisconsin productivity (Northcentral
           District) included 72% nest success (111 out 155 active
           territories produced 190 young) and productivity rates of 1.23
           young/active territory, and 1.71 young/successful nests.












                                                                          15

          Statewide, the number of active bald eagle territories increased
          from 418 in 1991 to 424 in 1992.

          Nest Abandonment- Nest Failures, and Nestling Mortality - 1992

               Although nest success on the Lake Sup erior shoreline was
          good in 1992, there were several territories, particularly on the
          APIS and in Chequamegon Bay, which were initially occupied but
          abandoned prior to egg laying. This abandonment of nesting
          effort is interesting due to the relatively close proximity of
          all nests involved. Several territories on the APIS (Basswood
          Is., Oak Is., Outer Is., Long Is., and Sand Point) had adults
          present and nest construction @ias documented. At four of the
          five territories a climber entered the nest, after the adults had
          abandoned, and observed fresh grass and the beginning of a nest
          cup but found no eggs (D. Evans, pers. comm.). In addition,
          nests at Madeline Is., Oak Point (AS-18)and Honest John Lake (AS-
          21) also had adults on territory but were not observed incubating
          during the spring flights. We were unable to enter these nests
          to ascertain their condition because they lie within the Bad
          River Reservation (Ojibway) who have not given the state
          permission to climb nests on their territory. All nests
          mentioned, with the exception of Outer Is., are in close
          proximity of Chequamegon Bay and the eagles would likely   use the
          bay as a primary feeding area. In the spring of 1992 ice cover
          was completely off of Chequamegon Bay on 23 April, later than
          normal (Tom Doolittle, NPS, pers. comm.). The eagles on the
          Chequamegon Bay also rely on commercial fisherman to provide food
          during the early spring months when nest initiation takes place;
          rough fish are thrown on the ice as nets are,retrieved and eagles
          frequently take these fish. In the winter of 1991-1992 there was
          a 56% reduction in commercial fishing effort through the ice on
          Chequamegon Bay (Tom Doolittle, pers. comm.), potentially
          reducing the available food to the eagles returning to the area.
          The water surrounding the peripheral islands (i.e. Devils is.,
          North-Twin Is., Outer Is., and Michigan Is.), on the other hand,
          did not freeze over last winter, allowing eagles on territory
          constant access to their prey base .(spawning burbot and lake
          herring) during the early nesting period (March-April). The
          reduction of a readily available food supply may have contributed
          to the reduced nesting effort on territories in close proximity
          to Chequamegon Bay in 1992.
               Three nests on the Lake Superior shoreline where direct
          observations had taken place failed at some point in the nesting
          season. The nest on the mouth of the Brule River (DU-9a) was
          observed empty on 9 May when the hatchlings were three days old.
          A climber entered the nest after it had been noticed empty and
          could find no evidence of young, nor evidence of any predator
          having been in the nest. The second confirmed failure was at BY-
          3b (York Island) where the adults were observed incubating as
          late as 27 May, > 40 days after they were first observed
          incubating. Analysis of the observation data indicated no











                                                                          16

           obvious lack of nest attentiveness or disturbance as incubation
           took place 98.3% of the time observed. Periodic checks were made
           of the territory and by mid-June no adults were seen. A climber
           entered the nest on 15 June and collected one addled egg for
           contaminant analysis. The final nest failure on the Lake
           Superior shoreline occurred in two stages. At nest BY-21 (Bark
           Bay) observations began on 18 May with two young in the nest. on
           7 June a climber entered the nest to collect nestling serum
           samples and noted that only one chick was present. A few eaglet
           feathers were on the ground and in a crotch in the tree about 3 m
           below the nest. The remaining eaglet was banded and a blood
           sample was taken. During the observation period the following
           week, 8 June, observers noted no chick activity in the nest and
           reported no prey deliveries. On 11 June the climber reentered
           ,the nest, found the eaglet was missing. After a short search the
           remains of the banded nestling were found on the ground within 30
           m of the nest tree. Remains included the head and beak, one foot
           (including the band), and numerous feathers, the rest of the
           carcass had been removed from the scene. The climber (D. Evans,
           raptor biologist with >20 years experience studying and banding
           bald eagles) concluded that the death was likely the result of
           mammalian predation. It was noted that the nest bowl had been'dug
           into, consistent with raccoon predation. The fact that the
           banding team@had visited the nest on 7 June and within 24 hours
           it had been predated seems to suggest an effect of some sort by
           the banding of the nestling.   It has been documented by video
           camera and direct observation at 5 nests this year that adults do
           not return to the nest until the day following nest entry by a
           climber. It is possible that the banding crew provided a scent
           trail to the nest tree and lack of adult attentiveness allowed
           predation to occur. It should be noted that one nestling had
           disappeared 10 days prior to the loss of the second chick. It
           should be emphasized that there-is no evidence that nest entry
           poses a general hazard to nesting bald eagles; in Wisconsin bald
           eagles have increased exponentially despite the fact that every
           nest in the state was entered and chicks banded from 1974-1989.
           However, individual nests may be placed at greater risk if the
           local predator population is high.. The fact that adults do not
           return to a nest until the day after a climber has been there
           also means that harsh weather can claim chicks if they are banded
           before they are able to thermoregulate (week 3-4) or during
           extremely inclement weather. A fourth nest failure occurred at
           North Twin Island (APIS), no observations were made at this nest
           site and the cause of failure is unknown.
               One nest, BY-15A (Fish Creek), contained two young at the
           beginning of observations, but only one remained on 28 May at the
           beginning of an observation period. The observations continued
           on a weekly basis with one chick in the nest when, on 25 June at
           0739, an observer noticed a fisher (Martes pennanti) climbing the
           nest tree. The adults returned to the territory scolding and
           circling around the nest. The fisher retreated down the tree.
           The remaining eaglet at this nest fledged on 9 July. It is











                                                                            17

           possible that the fisher was responsible for the loss of the
           nestling noticed missing on 28 May. observers have speculated
           that the expanding fisher population may be impacting raptors
           through nest predation in northern Wisconsin,'
                A minimum of 5 of 18 nestlings known to have hatched on Lake
           Superior died before fledging (28%). Only 1 nestling out of 11
           hatched was lost during the nesting season at the inland camera
           study sites (9-0o). -That mortality occurred at IR-9a on the Turtle
           Flambeau Flowage. It died between week 1-2 of age and was
           observed being out-competed for food by its more aggressive nest
           mate which was nearly twice its size. This nest had a much lower
           prey delivery rate (comparable to the rate at Lake Superior nest
           sites) than did the other inland nests, the adults were present.
           at the nest site much less frequently, and nestling mortality
           occurred at the nest on at least two other occasions in the past
           7 years.

                Nestling plasma, fish, and sediment contaminant levels
                Bald eagle nestling plasma', fish, and sediment PCB data are
           presented in Table 6. The sediment and fish PCB data presented
           is that available for feeding areas within 10 miles of sampled
           eagle nests on portions of the Wisconsin River, the Menominee
           River, the lower Fox River, Duluth/Superior Harbor, Lake Superior
           shoreline (including the Apostle Islands), and the Flambeau River
           (WDNR Water Resource Management, unpubl. data). Most sites
           sampled had known PCB contamination therefore there is not a wide
           gradient in fish/sediment/plasma PCB concentrations.
                The lower Fox River (Kaukauna) had the greatest
           concentrations of PCB in sediment and fish yet the nestlings had
           only intermediate plasma PCB concentrations (Table 6). Fish from
           the Flambeau River did not have detectable levels of PCBs (<0.2
           ppm) and nestling plasma PCB levels were low in samples collected
           nearby, however no detectable PCBs were found in sediment or fish
           within 10 miles of an eagle nest on the Wisconsin River yet the
           nestling plasma PCB concentrations was 147 ug/L. Fish collected
           near the outer islands of the Apostle Islands (primarily lake
           trout) had greater concentrations of PCBs than did fish from any
           other region along Wisconsin's Lake Superior shoreline, however
           Duluth/Superior harbor came in a close second (Table 6). The
           greatest concentrations of nestling plasma PCBs found along
           Wisconsin's Lake Superior shoreline occurred in some nestlings
           from the region (1991 North Twin and outer Island) however other
           nestlings had low plasma PCB concentrations. Of the fish
           sampled, only large lake trout had elevated PCBs, forage fish
           such as lake herring, lake whitefish, and burbot did not. A
           potential route of PCB exposure for these nestlings is that they
           are being fed commercial fishing waste (i.e. lake trout offal
           dumped overboard) or herring gulls and/or other colonial nesting
           birds or diving ducks known to have elevated PCB levels.
                Several caveats that need to be kept in mind when examining
           Table 6. First, the fish samples are primarily large game fish











                                                                          18

          fillets collected to assess human health concerns. We reported
          PCB values for fillets of fish >1811 as data for this size class
          existed for all sites which also had sediment PCB data. it
          should be noted that eagles rarely consume fish this large and
          generally consume the whole fish, not just the fillet. Therefore
          the fish PCB data presented does not represent the actual
          concentrations found in fish that eagles select at a given site.
          They may provide an index of a site's level of PCB contamination,
          however. The sediment PCB data should also be viewed cautiously
          as few sites were sampled extensively. The values presented
          represent the mean sediment PCB concentration of up to six cores
          taken at a given site (Fox River, Duluth/Superior Harbor) however
          most sites had only 2-3 cores taken. Because of varying rates
          sediment deposition/transport, a few samples may greatly over- or
          under- represent the contamination level at a given site.
               Therefore, to more accurately assess the relationship
          between nestling plasma PCB concentrations and that in the food
          it consumes, a determination of the size and type of prey they
          consume should be made, and representative samples of that size
          and species should be collected and analyzed. Sediment cores
          from several sites within a feeding area would also need to be
          taken if one wished to accurately assess the relationship between
          nestling plasma PCB concentrations and sediment contamination.

          Nestling plasma contaminant levels at Wisconsin RAP sites

               Bald eagle nestling plasma samples were obtained near three
          Wisconsin RAP sites 1990 - 1992. A nestling was sampled in 1991
          at DU-25, L. Pokegama, Douglas Co., and found to have plasma
          total PCB.concentration of 106 ug/L; p,pf-DDE was <IDL (2.5
          ug/L). This nest is near the St. Louis River/Duluth Harbor RAP
          site, approximately 4 miles upstream from the mouth of the St.
          Louis River. A nestling plasma sample and two addled eggs were
          collected in 1992 from MT-16, Hwy BB, Marinette Co., 4 miles from
          the Marinette RAP site. The plasma is currently being analyzed
          at Michigan State University. The addled eggs are frozen and
          archived with the WDNR. In addition, 3 nestling plasma samples
          were collected from OU-1a, Kaukauna, on the lower Fox River; 2 in
          1991 and 1 in 1992. This nest is 10 miles downstream from Little
          Lake Butte des Mortes, a primary s'ource of sediment PCBs for the
          lower Fox River and Green Bay RAP site. Plasma PCBs in 1991 =
          124 & 115 ug/L, p,pl-DDE = <IDL in both. The 1992 samples is
          currently being analyzed. An addled egg was also collected at
          OU-1a in 1ï¿½90, the Wisconsin Lab of Hygiene determined that the
          egg contained 38 ppm total PCB, 1 ppm DDE. An aliquot of the
          sample has been sent to the USFWS East Lansing office for AHH
          induction assay. Finally, a plasma sample was collected from a
          nestling at OC-4, Oconto River, in 1990. The sample is currently
          being analyzed. While the nest is 25 miles north of the Green
          Bay RAP site, these nestlings are also likely exposed to elevated
          level of OCs in the Green Bay estuary. Periodic sampling of
          nestlings and addled eggs at these nests and any new nests near











                                                                          19

          RAP sites (BR-1, Little Tail Point, Brown County has a subadult
          adult in residence in 1992, this nest is within 10 miles of the
          Green Bay RAP site) can provide a means for assessing contaminant
          trends and the result of remediation efforts at those sites.



                                   ACKNOWLEDGMENTS

          Bald eagle plasma analysis was conducted in the laboratory of Dr.
          John Giesy, MSU Pesticide Research Center under the supervision
          of W. Bowerman, Ph.D. candidate. Mr. Bowerman developed the
          protocol for bald eagle plasma sampling in the Great Lakes basin
          and trained WDNR personnel in the proper techniques. Data
          analysis was confirmed by D. Verbrugge, Head Chemist, and
          C.Vandervoort, QA Officer. D. Evans and R. Eckstein provided the
          expert climbing required to obtain bald eagle nestlings for
          sampling. P. Stefanek, T. Painter, K. Wollenburg (MSU), K.
          Warnke (University of Minnesota), and C. Dykstra (University of
          Wisconsin) collected nestling plasma samples and measured eaglets
          on the ground. Video equipment was lent to the project by the
          National Park Service (APIS) while additional equipment was
          purchased and personnel hired with a grant from the Great Lakes
          Protection Fund. J. Wilson and R. Eckstein (WDNR) mounted the
          video cameras at inland Wisconsin nest sites. J. Wilson and G.
          Kessler (WDNR Mercer) provided logistical support at the inland
          field sites. S. Stubenvol, L. Tesky, R. Eckstein (WDNR), and C.
          Sindelar conducted the aerial surveys of Wisconsin bald eagle
          nests in this study and C. Gieck (WDNR) provided historic
          reproductive data on the study nests which was collected by C.
          Sindelar. Numerous field assistants helped collect direct
          observational data including D. Kelner, G. Seebolde, D. Covell,
          K. Mooney, R. Schweickert. Volunteers - J. White, J. Plitz, N.
          Warnke, M. Parisi - also assisted at some observation sites. R.
          Brander, J. Van Stappen and T. Doolittle provided transportation
          and logistical support on the APIS. J. Amrhein (WDNR) provided
          data from the Wisconsin Fish Contaminant Monitoring program, L.
          Talbot (WDNR) provided Great Lakes sediment data, J. Stoyer
          (WDNR) provided sediment data from the lower Fox River/Green Bay
          Mass Balance Project, and J. Kreitlow (WDNR) provided sediment
          data from the Wisconsin River system. Financial support was
          received from the Great Lakes Protection Fund, the Wisconsin
          Coastal Management Program (NOAA), the National Park Service,
          WDNR Bureau of Research, and the WDNR Bureau of Endangered
          Resources Adopt an Eagle Nest Program.











                                                                          20


                                   LITERATURE CITED


          Bowerman, W. W., IV, K * D. Kozie, and M. Martell. 1989. Interim
               report: Bald eagle blood collection from nestling eagles in
               the Great Lakes Basin. Unpubl. rep. 6pp.

          Bowerman, W.W., D.A. Best, E.D. Evans, S.Postupalsky, M.S.
               Martell, K.D. Kozie, R.L. Welch, R.H. Scheel, X.F. Durling,
               J.C. Rogers, T.J. Kubiak, D.E. Tillit, T.R. Schwartz, P.D.
               Jones, and J.P. Giesy. 1990. PCB,concentrations in plasma
               of nestling bald eagles from the Great Lakes Basin, North
               America. In H. Fiedler and 0. Huttzinger, eds., 10th Intl.
               Conf. on Organohalogen Compounds, Bayreuth, Germany, Vol.
               IV, pp. 212-216.

          Bowerman, W.W. 1991. Factors influencing breeding success of
               bald eagles in upper Michigan. unpubl. M.A. thesis, N. Mich.
               Univ., Marquette, 119 pp.

          Frenzel, R.W. 1985. Environmental contaminants and ecology of
               bald eagles in southcentral Oregon. Oregon State
               University, Corvallis, unpubl. Ph.D. thesis.

          Garrett, M., R.G. Anthony, J.W. Watson, and K. McGarigal. 1988.
               Ecology of bald eagles on the lower Columbia River. Final
               Report to U.S. Army Corps of Engineers, Contract No. DACW57-
               84-C-0071, Oregon Coop. Wildl. Res. Unit, Oregon State
               Univ., Corvallis.

          international Joint Commission. 1989. Emerging issues--ongoing
               and emerging. (Appendix B).

          International Joint Commission. 1990. Draft: Final report of
               the ecosystems objectives committee. Unpubl. rep.

          Kozie, K.D. and R.*K. Anderson. 1991. Productivity, diet, and
               environmental contaminants in bald eagles nesting near the
               Wisconsin Lake Superior shoreline. Arch. Environ. Contam.
               Toxicol. 20:41-48.


          Mersmann, T. J, D. A. Buehler, J. D. Fraser, J. K. Seegar. 1992.
               Assessing bias in studies of bald eagle food habits. J.
               Wildl. Manage. 56:73-78.

          Michigan Department of Public Health. 1987. Analytical Method
               No.7, Cntr. Environ. Hlth. Sti., Epid. Stud. Lab., Lansing,
               Mi.

          Price, H.A., R.L. Welch, R.H. Scheel, and L.A. Warren. 1986.
               Modified multiresidue method for chlordane, toxaphene, and
               polychlorinated biphenyls in fish. Bull. Environ. Contam.
               Toxicol. 37:1-9.











                                                                         21

          Sprunt, A. IV, W.B. Roberstson Jr., S. Postapulsky, R.J. Hensel,
               C.E. Kroder, and F. J. Ligas. 1973. Comparative
               productivity of six bald eagle populations. Trans. N. Amer.
               Wildl. Nat. Res. Conf. 38:96-106.


          Webb, R. G. and A. C. McCall. 1973. Quantitative PCB standards
               for electron capture gas chromatagraphy. J. Chromgr. Sci.
               11:366-373.

          Wiemeyer, S. N., T. G. Lamont, C. M. Bunck, C. R. Sindelar, F. J.
               Gramlich, J. D. Fraser, and M. A. Byrd. 1984.
               Organochlorine pesticide, polychlorobiphenyl, and mercury
               residues in bald eagle eggs, 1969-79, and their
               relationships to shell thinning and reproduction. Arch.
               Environ. Contam. Toxicol. 13:529-549.



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                 Table 1

                 Project: Wisconsin DNR--Bald Eagle Plasma Analysis

                 Aquatic Toxicology Laboratory
                 201 Pesticide Research Center
                 Michigan State University
                 East Lansing, Michigan 48824                                                            Previous No.
                 Sample       Band Number     Year         Nest       Location                           pip$-DDE Total PCB
                 ------       ---------      ------        ------     ---------------------              ------      ------
                                                                                                            ng/ml      ng/ml
                         1    629-35441            1990    MT  8      GRAND RAPID POWER PLANT                [3.71          67
                         2    629-35437            1990    MT  8      GRAND RAPID POWER PLANT                  <IDL         54
                         3    629-35431            1990    MT  11     TWIN ISLAND                                10        105
                         4    629-35432            1990    MT  11     TWIN ISLAND                                23        213
                         5    629-35433            1990    MT  11     TWIN ISLAND                                13        136
                         6    629-35297            1990    OC  2      OCONTO R./BATTLE CREEK                      7         38
                         7    629-35298            1990    OC  3      W. PESHTIGO R.                             10         49
                         8    629-35208            1990    AS  12     N. TWIN                                    17         41
                         9    629-35332            1990    AS  16     BASSWOOD ISLAND                            43        135
                        10    629-35333            1990    AS  28     LONG ISLAND                                15         56
                        11    629-35334            1990    BY  23A    ORIENTA FLOWAGE                            30        102
                        12    629-35300            1990    IR  11     RICE LAKE                                  17         59
                        13    629-35315            1990    IR  33     TFF DAM                                     7         27
                        14    629-35319            1990    IR  25A    EAGLE ISLAND                           (2.6)          18
                        15    629-35321            1990    IR  34     SPRINGSTED LANDING                     (4.9]          47
                        16    629-35323            1990    IR  8C     MCGIVERS PASS                          (2.7)          32
                        17    629-35325            1990    IR  27     CEDAR LAKE                                  9         66
                        18    629-35327            1990    IR  19     OXBOW LAKE                                 12         35
                        19    629-35328            1990    IR  9C     MURRAYS LANDING                            11         52
                        20    629-35329            1990    IR  3B     BIG PINE LAKE                          (3.3)          19
                        21    629-35331            1990    IR  32     SAXON HARBOR                               36        124
                        22    629-35306            1990    JU  2      YELLOW RIVER                              7.3        197
                        23    629-35308            1990    NA  2A     BIG EAU PLEINE                         (4.8]          51
                        24    629-35311            1990    LI  7B     PINE RIVER                             [4.3)         103
                        25    629-35312            1990    LI  4A     GRANDMOTHER DAM                           6.4        147
                        26    629-31798            1991    MT  8      GRAND RAPIDS POWER PLANT               (3.0]          46
                        27    629-36322            1991    IR         STURGEON BAY                                6         56
                        28    629-36328            1991    IR  9C     MURRAYS LANDING                             6         47
                        29    629-36330            1991    IR         TWIN LAKES                                 77         75










                         Table I (Cont.)



                        30    629-36320             1991    IR 33      TFF DAM                                  [4.8]         46
                        31                          1991    SA 6B      BIG BLOCK                                <IDL          40
                        32    629-35388             1991    RU 5A      LADYSMITH                                <IDL          33
                        33    629-35389             1991    RU11       FLAMBEAU R.                              <IDL          27
                        34    629-35377             1991    OC  1      ARCHIBALD L.                               14          37
                        35    629-35376             1991    OC  3      WAUPEE FLOWAGE                               a         19
                        36    629-35373             1991    SH  1      SHAWANO L.                               <IDL          30
                        .37   629-35372             1991    MT  9      HIGH FALLS                               <IDL          24
                        38    629-35369             1991    MT  2      L. NOQUEBAY                                  8       330
                        39    629-35392             1991    OU  I      KAUKAUNA/FOX #1                          <IDL        124
                        40    629-35394             1991    OU  1      KAUKAUNA/FOX #2                          <IDL        115
                        41    629-36340             1991    DU  2      AMNICON RIVER                            <IDL          79
                        42    629-36331             1991    AS, 16     BASSWOOD ISL.                              24          73
                        43    629-36333             1991    AS  15     OAK ISLAND                                 20        114
                        44    629-36334             1991    IR  32     SAXON HARBOR                               16          66
                        45    629-36336             1991    DU  9      BRULE MOUTH                                15          70
                        46    629-35395             1991    DU  25     L. POKEGAMA                              <IDL        106
                        47    629-36338             1991    BY  21     BARK RIVER                               <IDL          85
                        48    Not on Vial           1991    AS  26     DEVILS ISL.   #1                           11          43
                        49    Not'on Vial           1991    AS  26     DEVILS ISL.   #2                           10          83
                        50    Not on Vial           1991    AS  12     N. TWIN ISL.                               42        201
                        51    Not an Vial           1991    AS  3      OUTER ISLAND                               56        198
                        52    629-36318             1991    VI  89     EAGLE RIVER                              <IDL          24
                        53    629-36326             1991    ON  8      SWAMP CREEK                              <IDL          59
                        54    629-36316             1991    ON  25     BACKWATER BAR                            [4.4]         31
                        55    629-35399             1991    LI  7      PINE RIVER                                   8       118
                        56    629-35365             1991    MA  3      HALF MOON L.                             <IDL        156
                        57    629-35363             1991    WO  2      SCHMIDT SLOUGH                           <IDL        187
                        58    629-35360             1991    PO  3      STEVENS POINT                                6         31
                        59    629-35359             1991    JU  2      YELLOW RIVER                             <IDL        100
                        60    629-35357             1991    AD  1      CASTLE ROCK                                  9       277

                  (n]   Below level of   quantification (LOQ)     for method
                  <IDL     Below instrument detection level      (IDL) of 2..5 ng/ml
                  IDL   2.5 ng/ml, p,pl-DDE; 5 ng/ml, PCBs
                  LOQ   5 ng/ml, p,p'-DDE; 10 ng/ml, PCBs for 2 ml plasma extracts










              Table 2.      Wisconsin     1990-91   nestling     bald  eagle    plasma    contaminant
                            concentrations, by location.



                                      total PCB (ug/L)                  p,pl-DDE (ug/L)

                                    ------------------------        -------------------------
                                  % with      Geo-                  % with      Geo-
                                  Detect-     Metric                Detect-     Metric
              Location      N       able      Mean     Range         able       Mean     Range


              1.            16      100        84      41-201          81         16     <IDL-56


              11.             1            Sample analysis underway at    MSU

              111.          13      100        47      18-75          100          6     3-77


              IV.           13      100        46      19-330          77          7     <IDL-23


              V.            11      100       125      31-277          58          4     <IDL-9


              VI.             7     100        33      24-59           17       <IDL     <IDL-5



                    Location   I -  <8 km from Lake Superior
                    Location   II-  <8 km from Lake Michigan
                    Location   III  - counties adjacent to Lake Superior & > 8 km inland
                    Location   IV - counties adjacent to Lake Michigan & > 8 km inland
                    Location V      inland counties, historic point source discharge
                    Location VI - inland counties, no historic point source discharge








             Table 3. Mean concentrations and ranges (ug/L) of FCBs and DDE from nestling bald

             eagles in the Great Lakes Basini..

                                    Total PCBs                     DDE


             Nesting Area      n    Mean            Range           Mean        Range




             Great Lakes       42    183.3        33.0-520.0        60.9        13.0-306.0


             Interior          79     23.7        5.0-200.0         20.0         2.0-193.0




               Bowerman et al. 1990









                Table 4. Mean concentrations and ranges (ug/L) of PCBs and DDE in whole blood of nestling bald eagles from

                Oregon and Washingtonl. Concentrations have been corrected by a conversion factor of two to be equivalent to

                blood plasma values.

                                                      Total PCBs                      DDE


                Breeding Area                   n     Mean         Range        Mean         Range




                Lower Columbia River            14     129.0       0.0-351,0           98.0         66.2-130.5

                Outer Klamath Basin             24       22.0      0.0-580.0           44.0         nc3

                Upper Klamath Lake              17        0.0      nd 2-280.0          46.0         nc3

                Cascade Lakes                   34        0.0      nd 2-280.0          20.0         nc3



                1 Frenzel, 1985; Garrett, et al. 1988.

                2 nd - not detected in sample.

                3 nc @ not given in citation.









                         Table 5      1992 Blood Collection Territories in Wisconsin
                    Territory         Date          Band         Weight (kq)         Sex           Blood
                                                                                                Coll.-(cc)

                       WO-2c         5/29/30       629-36342           NW             F               10
                       =-2a          5/29/92       629-36344          3.65            m               10
                       A.D-I         5/29/92       No Band'            NW           Unk.              10
                       OU-1a         5/30/92       629-36346          3.66            F               10
                       OU-1a         5/30/92       629-36347          3.43            F               10
                       MT-16         5/30/92       629-36349          2.60            F               9
                       SA-14a        6/2/92        629-36:YlO         2.94            F               10
                       WA-34         6/2/92        629-36312          3.05            m               10
                       SA-60e        6/2/92        629-36313          3.91            F               10
                       SA-65         6/2/92        629-36314          4.30            F               9
                       BA-3a         6/3/92        629-36315          4.72            F               10
                       WA-11         6/3/92        629-37362          3.44            m               10
                       VI-65b        6/4/92        629-36356          3.57            m               11
                       BY-23         6/7./92       629-36360          3.40            m               10,
                       BY-21         6/7/92        629-363622         3.35            m               10
                       AS-25         6/7/92       .629-36363          4.30            F               10
                       BY-15a        6/7/92        629-36365          4.25            F               10
                       IR-9c         6/9/92        629-36369          3.81            F               10
                       IR-33         6/9/92        629-36370          4.15.           F               10
                       DTJ-18        6/10/92       629-36358          4.65            F               10
                       SA-27a        6/10/92       629-36373          3.80            m               10
                       DU-25b        6/11/92       629-36.375         4.50            F               10
                       IR-32a        6/12/92       629-36377          3.20            m               10
                       VI-61C        6/13/92       629-36366'         3.15            m               10
                       VTL-57        6/13/92       629-36380            NW                            10
                       DU-02g        6/16/92       629-36381          4.35            F               10
                       DU-24a        6/17/92       629-36382          4.34            F               10
                       AS-24a        6/18/92       629-36384          3.60            m               10
                       DU-14a        6/18192       629-36385          4.20            F               10
                       IR-35a        6/19/92.      629-36362  2       3.15            m               10
                       AS-20c        6/22/92       629-36387          3.85            m               10
                       AS-25         6/24/92       629-36389          4.65            F               10
                                     6/27/92       629-36386          3.90            m               10









                This eaglet was captured on the ground af ter it had
                fledged, the bander was not present.          Sample and
                measurements were taken.

                'This band was first placed on the eaglet at  nest BY-21.
                Several days after banding the eaglet 'was f ound dead
                below the nest tree. The band was recovered and placed
                on the chick at nest IR-35a (Blair Lake).










           Table 6. Wisconsin bald eagle nestling plasma, sediment, and
           fish fillet total PCB content, by location.

                 7-------- -------------------------------------------------
           Location         Nestling plasma       Fish (>18")     Sediment PCB
                            total PCB (ug/L)      fillet PCB      (0-5 cm)
                                                  ug/g fresh      ug/g dry
                                                  Mean,(range)    Mean (range)
                                               -------------------------------
           Duluth/          106                   0.7 (<0.2-1.1) 0.2 (0-1-0.6)
           Superior
           Harbor

           L. Superior          79                0.3 (<0.2-0.6) NM
           (Douglas)

           L. superior      85, 102               ND              NM
           (Bayfield)

           L. Superior      56, 135               0.2 (<0.2-0.5)  NM'
           (inner APIS')

           L. Superior      41, 43, 83,           1.3 (<0.2-5-8)  NM
           (outer API S3)   198, 201

           L. Fox River/    115, 124              2.5 (0.4-9.8)   2.6 (0.5-4.8)
           Kaukauna
           Menominee 4R./   213, 136, 105         1.6 (0.3-3.0)   0.1 (0.1-0.2)
           Marinette

           Wisconsin R.     100, 187, 197,        .0.6 (0.3-1.3)  NDI
           (Castle Rock/    277
           Petenwell)

           Wisconsin R.     31                    0.8 (0.3-1.9)   0.2
           (Stevens Pt)

           Wisconsin R.     156                   0.3             0.6 (0.4-0.8)
           (Mosinee-
           L. Du Bay)

           Wisconsin R.     103, 118              1.0             NM
           (below Merril)

           Wisconsin R.     147                   ND              ND
           (below Toma-
           hawk)

           Flambeau R.      27, 33, 37            ND              NM











          Table 6 (cont.)


          ND - none detected
          NM - not measured
          1  - Basswood Island, Oak Island
          @2 - samples currently under analysis with US Geological Survey
          3  -Devils, N. Twin, & Outer Islands
          4  - Plasma sampled at Twin Islands, 10 miles upstream from fish
               and sediment samples (Marinette)
          5
               sediment collected at mouth of tributaries, none collected
               in deposition zones



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               Figure I


                      NESTLING EAGLE PLASMA SAMPLE LOCATIONS
                                                    1990-1991


                                                                               0 Sampled In 1990

                                                                                    Sampled In 1991



                                                                       3" Detailed Map











                               Figure 2




                                                   LAKE SUPERIOR REGION

                                      NESTLING EAGLE PLASMA SAMPL                                                                              ES

                                                                               1990-1991




                                                                                                                                   Outer 1.
                                   E      Sampled            in 1990                             Devil's 1.      North Twin 1.
                                          Sampled in 1991




                                                                                           York 1.                                   Oak 1.





                                                                                                                                                       Michigan 1.
                                                                                                                                    Basswood L


                                                                                                                                       Long Island











                                                                                                                                 Iron
                                   Douglas                               Bayfleld                                                            MEN
                                                                                                                                                **N
                                                                                                                                                E
                                                                                                           Ashland                                    N*N










                                 Figure 3



                                             WISCONSIN BALD EAGLE RESEARCH SITES
                                                                                A               1992
                                                                          A

                                                              SW@id                                                       N serum samples
                                               Douglas                              A,,_A,
                                                                                  nd                                             observation blinds


                                                                                                                                 camera nests

                                                                                                    Was
                                                  Washburn Sawyer


                                                                                  Price                               Forest     Florence
                                  Burned                                                         Onsili    m
                                        Polk            *I                                            *    *
                                                             Rusk                                                                   Marinette
                                                                                                 Lincoln       Langlad.a
                                                                            Taylor                                            Cconto
                                 St. Croix     Dunn           Chippewa                                                         I
                                                                                             Marathon                    enaminee
                                                                            Clark                                                                           Doo

                                Pierce                    EAu Claire                                                             Shawan

                                                                                          Wood      I-rtag*        Waupaca
                                            SO    Buffalo    9V                                                                Cutagamie
                                                                               Jackson                                                    Brown
                                                                                          Juneau I Adams Wausham            nnebago      umall Manit
                                                                            lonroe

                                                                   eras"
                                                                                                           Marqu Greenj
                                                                                                           BUD      Lake                   Shebayg


                                                                   Vernan                                                  Fond Du Lac
                                                                                  FUchiand  Sauk          C41umbia      Dodge          ashing@oz
                                                                                                                                       on      auk
                                                                    Crawford                           Dane
                                                                      Grant            IOW4                             Jefferson I Wauii--n4LU@
                                                                                       Lafayette                              walwo      Racine
                                                                                                                                            Kenash










                           Figuy.e 4


                        Wisconsin Bald Eagle Nestling Plasma PCB Concentrations
                                                     By Location (1990-1991)

                                 350



                                 300


                              0)
                              :3 250



                              u  200        0


                              @2 150
                              M
                              E
                              In 100
                              M
                              FL

                                   50



                                     0
                                        <8km from     >8km from            >8km from           Below Historic No Historic
                                        L. Superior   L. Superior          L. Michigan          Point Source   Point Source
                                               Great Lakes Counties                             Inland Counties











                            Figure 5


                     Wisconsin Bald Eagle Nestling Plasma DDE Concentrations
                                                    By Location (1990-199 1

                                  60



                                                                (77)
                                  50


                               CD
                                @:' 40
                                LU


                               C1 30
                               CL -


                               E
                               Ch 20         0
                               CO


                                  10



                                     0
                                        <8km from      >8km from             >8km from            Below Historic  No Historic
                                          L. Superior   L. Superior          L. Michigan          Point Source    Point Source
                                                 Great Lakes Counties                             Inland Counties








                     Figure 6


               PLASMA PCBs VS. EAGLE PRODUCTIVITY
                                        WISCONSIN 1985-92
           2.5





             2 -





           1.5 -





         z

         0



           0.5 -





             0
              0         50       100       150      200       250      300       350
                                   PLASMA TOTAL PCBs (ug/L)




                               PLASMA PCBs VS. YOUNG/NEST ATTEMPT

                                         Regression output:
                               Constant                      1.253711
                               Std Err of Y Est              0.484551
                               R Squared                     0.025469
                               No. of observations                 48
                               Degrees of Freedom                  46

                               X Coefficient(s) 0.001219
                               Std Err of Coef. 0.001112









                 Figure 7


               PLASMA DDE VS. EAGLE PRODUCTIVITY
                                       WISCONSIN 1985-92
            2.5





             2





            1.5


        cn




        :D
        0      EME
        4k
            0.5                              M         M

                                                                      M



             0               f      I
              0      10     20      30     40     50      60     70     80      90
                                    PLASMA p,p'-DDE (ug/L)




                                PLASMA DDE VS YOUNG/NEST ATTEMPT

                                         Regression Output:
                                Constant                     1.536152
                                Std Err of Y Est             0.434254
                                R Squared                    0.217284
                                No. of Observations                48
                                Degrees of Freedom                 46

                                X Coefficient(s) -0.01445
                                Std Err of Coef. 0.004043








                     Figure 8



                      PLASMA PCBs VS. PLASMA DDE
                                        WISCONSIN 1990-91
           90


           80  -


           70  -


           60  -


           50  -


           40  -


           30  -


           20  -
                      0  H%
                     0
           10  -


             0
               0       50        100       150      200       250      300       350
                                   PLASMA TOTAL PCBs (ug/L)




                               PLASMA PCBs VS.  PLASMA DDE

                                         Regression output:
                               constant                      71.83647
                               Std Err of Y Est              63.69064
                               R Squared                     0.071792
                               No. of Observations                 60
                               x coefficient(s) 1.226053
                               Std Err of Coef.      0.57887











                   Figure 9



                  LAKE SUPERIOR VS. INLAND BALD EAGLE
                                 PREY DELIVERY RATES
                                    PREY DELIVERIES/1 6 HOURS
                  fa lo
                  0                INLAND                  LAKE SUPERIOR


                  0                                                           -Oesnoje- MoOtTALSTY
                  3:
                  (D 6-
                  CA
                  LLJ
                  jE   4
                  LLJ


                       2
                  cl


                                                                      All
                  Uj   0
                  m      CARROL HHL   TFD   LSG    ML  PARK   R@EF  FISH  SAX  MICH
                        t
















                                                TERRITORY











                                Figure 10

                                          1992 Bald Eagle Prey Type
                                                Wisconsin Inland vs. Lakeshore

                              .80


                              70


                              60


                              50


                         CD
                              40


                         2    30


                              20


                              10


                                 0
                                     Fish, Mammal     Bird    Reptile  Unk                       Fish  Mammal     Bird   Reptile   Unk


                                                 Inland (n=694)                                             Lakeshore (n=92)











                            Figure 11
                          1992. Bald Eagle Prey Size Classes
                                           Wisconsin Inland vs. Lakeshore


                           60




                           50



                       .a) 40
                       a)
                       Z



                       a)
                           30
                       CL

                       0

                       C

                       CL  20



                           '10



                             0 -
                                  0-6"   6-12" 12-18"   > 18" Unknown                  0-6"   6-12"  12-18"  >18" Unknown


                                           Inland (n=694)                                          Lakeshore (n=92)













                              Figure 12


                            WISCONSIN BALD EAGLE PRODUCTIVITY (1983-1992)
                                        LAKE SUPERIOR VS. INLAND NEST SITES
                                                    YOUNG PRODUCED/OCCUPIED TERRITORY
                                 1.6



                              0
                                                                                  . ............. ..... ..................
                                                                                .......... .





                                 1.2
                             13







                                 0.8

                              :3                NORTHCENTRAL DISTRICT LAKE SUPERIOR    >20 MILES INLAND
                              0                                                      (ASH., BAYfOU.G. CO.)
                                 0.6                  .......... 0. ........



                                 0.4
                                                                                                        v




                                       83        84 85              86        87       88 '89              90        91        92
                                                                                 Year



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             122392 03:54PM FROM MSU-FISHERIES                                                                                     P03





                 MICHIGAN STATE UNIVERSITY


                 PESTICIDE RESEARCH CENTER                                                                EAST LANSING MICHIGAN 48824-1331
                       TELEPHONE (517) 353-6376                                                                             FAX: (517) 313-3596




                 December 23, 1992


                 Dr. Micheal Meyer
                 Wisconsin DNR
                 Bureau o8f Research
                 1350 Femrite Road
                 Monona, WI 53716

                 Dear Dr. Meyer,
                          I am enclosing the Quality Assurance Report for the Wisconsin
                 DNR--Bald Eagle Plasma Analysis.                                    If I can be of further
                 assistance, my number is (517) 336-2028.

                                                             Sincerely,
                                                                  
                                                              
                                                            Christine        Vandervoort










































                                                    MSU is an Affirmative Action/Equal Opportunity Institution




        122392 03:54PM FROM MSU-FISHERIES                                    P04





           MICHIGAN STATE UNIVERSITY


           PESTICIDE RESEARCH CENTER                               EAST LANSING MICHIGAN  488244-1311
           TELEPHONE (517) 353-6376                                      FAX:  (317) 353-5598


                                 QUALITY ASSURANCE REPORT

           STUDY;                Wisconsin DNR--Bald Eagle  Plasma Analysis
                                 Cooperative Research with Wisconsin DNR  and
                                 Michigan State University
                                 Aquatic Toxicology Laboratory




           STUDY DIRECTOR:       Dr. John P. Giesy
           STUDY FACILITY:       Michigan State University
                                 Aquatic Toxicology Laboratory
                                 E. Lansing, 8qM8qI 48824

           DATE OF REPORT:       December 23, 1992
                I have reviewed the PCB and  DDE  anayltical chemistry work done
           on the Bald Eagle Plasma' collected In Wisconsin.  The work was
           performed in Dr. John Giesy's laboratory at Michigan State
           University. The following is my opinion of the study mentioned 
           above.
           

                The analysis was performed according to the SOP for the 
           analysis by personnel of appropriate training and experience. The
           data analysis was done by William Bowerman and confirmed by Dave
           Verbrugge, Head chemist for Dr. John Giesy. My review of data
           analysis agrees with the data presented.

                The study appears to have been performed as stated in the  work
           plan. If you have any questions I may be contacted at (517)  336-
           2028.

           Report written by:    Christine Vandervoort, QAO
                                 Pesticide Research Center
                                 Michigan State University
                                 E. Lansing, MI 48824



























                                                                                 SOP NO.


                                         Michigan State University
                                         Pesticide Research Center
                                       Aquatic Toxicology Laboratory



                                       SaWEDARD OPZRAT33FG PROCZDURZ


                      ANALYSIS OF ORGANOCHLOR    PESTICIDE15 "d PC33s 13 BIRDS' P!Lal@"


                                                                 Prepared by: Miguel A. Mora
                                                                               Dave Verbrugge


              I. SCOPE


                    The scope of this method is to determine the concentrations of
                    organachlorine pesticides (OCs) and polychlorinated biphenyls (PCBs) in
                    plasma of wild birds. A list of the compounds that can be determined by
                    this method and the individual detection limits are given in Table 1.
                    The extraction and cleanup procedures are adapted from Sonzogni et al.
                    (1991), Burse et al. (1990), Schmitt et al. (1985), and SOP from
                    Michigan Department of Public. Health (1987) with some modifications.
                    The method Is precision is within 10% and the accuracy >90%. Total. PCBs
                    are reported as a mixture of Aroclors 1242, 1248, 12S4, and 1260.
                    Chromatograms of standard mixtures are given in Appendix. The
                    instrument detection limit (IDL), method detection limit (MDL), and
                    method quantitation limit have been determined as described in Taylor
                    (1989).


              I I . REFX1tWFCZS


                    Burse,V.W   S.L., Read, M.P. Korver, P.C. McClure, J.P. Donahue, and L.L.
                          Needham. 1990. Determination of selected organochlorine
                          pesticides and polychlorinated biphenyls in human serum. J. Anal.
                          Toxicol. 14:137-142.


                    Michigan Department of Public Health. 1987. Analysis of blood for
                          polychlorinated and polybrominated biphenyls, and chlorinated
                          hydrocarbon pesticides. Analytical Method No 7. Lansing,
                          Michigan.

                    Monro, A.M. 1990. Interspecies comparisons in toxicology: The utility
                          and futility of plasma concentrations of the test substance. Reg.
                          Toxicol. Pharmacol. 12:137-160.


                    Schmitt, C.J., J.L. Zajicek, and N.A. Ribick. 1985. National pesticide
                          monitoring program: Residues of organochlorine chemicals in
                          freshwater fish, 1980-81. Arch. Environ. Contam. Toxicol. 14:225-
                          260.


                    Sonzogni, W., L. Maack, T. Gibson, D. Degenhardt, H. Anderson, and B.


                                                     1









                           Fiore.   1991. Polychlorinated biphenyl congeners in blood of
                           Wisconsin sport fish consumers. Arch. Environ. Contam. Toxicol.
                           20:56-60.


                     Taylor, J.K. 1989. Quality Assurance of Chemical Measurem    ents. Lewis
                           Publishers, Inc., Chelsea, Michigan, 328 pp.


              III. SUMMARY


                     This method permits the separation of organochlorine pesticides and PCBs
                     from small volumes of birds' plasma. One ml plasma fractions are
                     denaturized with methanol, extracted with a 1:1 mixture (v/v) of hexane-
                     ethyl ether and cleaned up with mixed solvents in florisil and silica
                     gel columns. The florisil and silica gel fractions are analyzed by gas
                     chromatography with electron capture detector (GC-ECD). OCs and PCBs
                     are confirmed by GC/MS in 10% of the samples. Average recoveries for a
                     mixture of 6 pesticides were 77.5% (88% without dieldrin) and       % for
                     PCBs.


              IV.   SZGNIFICANCE AND USE


                     The determination of org anachlorine pesticides and PCBs in plasma of
                     wild birds is of Importance for the understanding of the relationships
                     of the distribution of xenobiotics between plasma and body tissues.- The
                     plasma reaches many target organs and xenobiotics are removed from the
                     plasma as this goes through a given tissue (Monro 1990). By being able
                     to determine concentrations of organochlorine pesticides and PCBs In
                     plasma we can accomplish the following: first, we can take samples of
                     individual species without having to kill them; and second, the same
                     individuals can be sampled over time allowing us to determine seasonal
                     variations and bioaccumulation of contaminants in marked individuals.


              V.    T r5l       S


                     There are components in plasma that may produce some interference. This
                     can be avoided by following an adequate cleanu procedure.
                                                                     T
                     interferences will be detected by comparing the environmental samples
                     with periodical runs of chicken plasma blanks. Prior to initiating the
                     studies the lab facility will be carefully cleaned to reduce
                     contamination risks. Chicken plasma (obtained from the chicken farm
                     MSU) will be used as control and for recovery experiments.


              VI. APPARATUS


                     Gas chromatograph, Perkim Elmer, model 8500, with electron capture
                     detector (ECD) with  63 Ni foil at 3500C. Coli-1: DB-1 fused silica
                     capillary column (J & W Scientific), 30 m x 0.25 mm i.d., 0.25;Lm film
                     thickness. Injector in splitless mode. Septum purge set at 3-5 ml per
                     minute, temperature at 220*C. Carrier gas: helium at 5 psig, flow rate
                     of @ 1 ml per minute. Makeup gas nitrogen at 45 psig. Total flow rate
                     of @ 50 ml/min. Autosampler Perkin Elmer 8300. Data (retention times
                     and area percentages) are transferred directly to a microcomputer.

                                                       2












              VI I. REAGENTS AND NaTERIALS


                    A.    Reagents: Methanol, hexane, diethyl ether (peroxide free),
                          petroleum ether, isooctane, benzene, and acetone; Burdick and
                          Jackson, Baxter, Muskegon, Michigan. All solvents used are of
                          high purity or pesticide grade quality.


                    B.    Sodium sulfate, anhydrous, granular and powder forms.   Rinse with
                          hexane or methylene chloride in a buchner funnel 0before use. Let
                          air dry for a while, then dry in the oven at 130 C for at least 24
                          hr before use. May keep stored at 130  0C.

                    C.    Glass wool. Rinse glass wool with methylene   chloride orbexane
                          for at least 24 hr before use.

                    D.    Florisil, 60/80 0mesh, PR grade, Floridin Co., Pittsburgh, PA.
                          Activate at 130 0C for at least 48 hr before use. Keep stored in
                          the oven at 130 C.
                    E.    Silica gel 60, 70/230 mesh. Activate at 130  0C for at least 24 hr
                          before use. Store at 130  0C.

                    F.    Glassware. All glassware is washed with liquinox detergent
                          rinsed with tap and deionized water, then rinsed with acetone and
                          hexane before use.


                    G.    Reference standards.


                           1.   Pesticide matrix spike, (3/90) catalog # 32018, Lot
                                A000071, Restek Corporation, Bellefonte, PA.
                           2.   Certified reference material, PCBs (aroclor 1260) in human
                                serum, lot # SRM1589; National Bureau of Standards (MBS),
                                Gaithersburg, MD.
                           3.   Internal standard, PCB 30, obtained from Acustandards.
                                Stock and working solutions were prepared in our lab.
                           4.   Aroclors 1242, 1248, 1254, and 1260, obtained from Dr.
                                Zabik, Pesticide Research Center, MSU, originally obtained
                                from Monsanto Co. Stock and working solutions were prepared
                                in our lab.
                           S.   Chlorinated hydrocarbon pesticides: Analytical reference
                                standards obtained from U.S. EPA, Quality Assurance
                                Division, Research Triangle Park, NC. Stock and working
                                solutions and mixtures were prepared in our lab.
                           6.   Bovine serum and plasma reference material, obtained from
                                Department of Public Health, Lansing, Michigan (MDPH).
                           7.   EPA human plasma for blind analysis (interlab. studies),
                                obtained from MDPH.


             VIII. HAZARDS AND PRECAUTIONS


                    Some of the solvents used are flammable and explosive. Solvents should


                                                     3









                   be always used under the hood and away from fire. Use of lab coats and
                   eye protection goggles is important. In case of a spill, skin contact
                   or inhalation problems, follow specifications in material safety data
                   sheets (MSDS). Handling and storage precautions should follow the
                   recommendations described in the respective MSDS. Waste should be
                   collected and disposed properly according to HSU ORCBS indications.


             IX. SAMPLING AND SAMPLE PREPARATION


                   Blood from the brachial vein of fish-eating birds (caspian terns,
                   double-crested cormorants and bald eagles) was collected in heparinized
                   tubes according to specified procedures. Each sample was marked with
                   bird's common name, location, date and band number (if banded). The
                   samples were either centrifuged in the field (10 min at 3000 rpm), or
                   placed in a refrigerator (4 0C) and centrifuged within 48 hours. The
                   plasma was separated from the red blood cells and stored in the freezer
                   until chemical analysis.


             X. PREPARATION OF APPARATUS


                   Prior to use the instrument performance is mostly determined from
                   previous runs. Check pressures of He at 5 psig, N2 at 45 psig. If
                   gases are not on, turn make-up gas on, then turn auxiliary pressure
                   control knob to suggested psig. Percent saturation is adjusted to.0.9%.
                   If the baseline is appropriate, then we can assume that the working
                   conditions are optimal. Column performance should be determined from
                   previous recent runs, and by injecting standards before the autosampler
                   run. The autosampler is loaded and the QC run is set in the computer.
                   Set computer to receive information from each run, then set output (GC)
                   to external device.


             XI. CALIBRATION AND STANDARDIZATION


                   Availability and use of appropriate standards: our pesticide laboratory
                   standards have been evaluated with the use of a certified pesticide
                   matrix spike, catalog # 32018, lot # A000071, Restek Corporation,
                   Bellefonte, PA, The relative response factors obtained for the two sets
                   of standards were withim


                   The performance of the GC will be monitored daily by measuring the
                   response and retention times of several calibration mixes. The number
                   of theoretical plates will be calculated using two compounds, C20-ATA
                   and 2,4,6-trichlorobiphenyl (IUPAC #30). The ratio of the theoretical
                   plates (#30/C20) will be used to monitor the condition of the column. A
                   record of the retention times, peak responses, theoretical plates, and
                   peak shape will be kept in the GC Log book. If the theoretical plate
                   ratio changes by > Y25% from its mean value, or if serious column
                   deterioration is observed, the column may be replaced if the situation
                   cannot be corrected. If the retention time of any internal standard
                   changes by > 0.5 min from its mean value, the system will be checked and
                   corrected as required.



                                                    4









                    The linear range of the GC will be established for pesticide mixtures
                    and for a 1:1:1:1 mix of Aroclors 1242, 1248, 1254 and 1260 using a
                    performance relative response factor (PRRF)., The PRRF is defined by the
                    equation (ex. for aroclor);


                    PRRF = ARtotaiarea*ISTI)conc/ARconc*ISTDarea


                          AR-- Aroclor Mix     ISTD= Internal Standard
                          total area= sum of peak areas for the araclor mix
                          area= peak area for ISTD
                          conc= concentration in ng/jUl


                    The PRRF is specific for each OC or for a 1:1:1:1 mixture of Aroclors,
                    and is used only to monitor instrument performance. The PRRF will be
                    constant over the linear range of the detector. Constant is defined as
                    Y3% from the mean value for the PRRF. This range will encompass a
                    minimum of 1.5 orders of magnitude using a minimum of 3 concentrations.
                    The target operating linear range will be 5, 2.5, and 0.15ng of Aroclor
                    mix injected, and 0.25, 0.1, and 0.01 ng for OCs. Once the linear range
                    has been established, an individual standard solution for each of the
                    mixtures will be. chromatographed. These chromatographs will be used as
                    templates for pesticide mixture's and the Comstar PCB pattern recognition
                    program. The integrity of the template will be checked by daily
                    injection of pesticide mixtures and a 1:1:1:1 Aroclor performance
                    standard. The absolute concentration of the performance standard will
                    be adjusted to the linear range of the instrument. The calculated
                    concentration of the mix should be @710% of the expected value.

                    Calibration checks will be run at the beginning and end of a sample setf
                    where a set is approximately 10 samples. If the concentration of the
                    standard mix is outside of the 10% range the template will be
                    rechromatographed prior to further sample analysis.
                    A log of the relative response factors (RRF) for the individual Aroclors
                    will also be maintained as a check of the GC performance over the course
                    of the study. The.RRF is defined by the equation:

                    RRF -@ ARtataLarea*ISTDcanc/ARconc*ISTDarea


                          AR= Individual Aroclor       ISTD= Internal Standard
                          total area= sum of peak areas for the aroclor
                          area= peak area for ISTD
                          conc= concentration in ug/;Al

                    If the RRF for a given pesticide or aroclor changes by > 10% from its
                    mean value, the instrument will be checked and the appropriate
                    maintenance (le. bakeout, clean detector, etc.) will be completed before
                    prior continuing with the analyses. The standards should be re-
                    chromatographed and new templates prepared.






                                                    5













             XII* PROCZDURE


                  A. SAIOLE extraction.


                      1)  Transfer 1 ml of plasma to a 10 ml test tube with teflon cap.
                          Record also plasma weight by difference from test tube weight.

                      2)  Add 0.5 al methanol and vortex for about  five seconds.


                      3)  Extract with 5 ml of hexane-ethyl ether (1: 1, v/v) by agitation in
                          a burrel-wrist action shaker for 10 min.


                      4)  Transfer extract to centrifuge tube and centrifuge at 2000 rpm for
                          5 min. Transfer extract to a second centrifuge tube to combine
                          the extract volumes and further evaporation.

                      5.  Repeat extraction procedure (steps 3 and 4) twice (three times
                          total).


                      6.  Add 0.5 ml of isooctane, then concentrate extract to 0.5 ml in a
                          rotary evaporator or N-evap on a warm water bath.


                  B. FLORISIL cleanup and fractionation.

                      1   Prepare columns by placing 1 cm of granular anhydrous Na2SO4 On
                          glasswool in a 1 cm. x 30 cm i.d. chromatography column fitted with
                          a 250 ml reservoir. Add five grams of 60/80 mesh Florisil and top
                          with another 1 cm layer of sodium sulfate.


                      2)  Wash each column with 20 ml of petroleum ether and discard
                          resulting'effluent.


                      3)  When petroleum ether reaches the top of the Na2SO4 layer, add the
                          concentrated extract (approx. 0.5 al) and allow it to drain into
                          the col-n. Rinse the flask at least three times with @ 1 =I of
                          petroleum ether each time. Transfer the rinses into the column
                          and discard the eluent resulting from loading and rinsing.

                      4)  Wash the column walls with 5-ml of 6:94 ratio of diethyl
                          ether4petroleum ether and collect the eluent in a 250 ml round-
                          bottom flask. When the solvent reaches the top of the Florisil
                          add another 30 =1 of the 6:94 solvent and continue collection.
                          Set this fraction aside for silica gel fractionation.

                      5)  Repeat the above procedure using a 20:80 ratio of diethyl
                          ether:petroleum other in place of the 6:94 solution and collect
                          the eluent from the 5 ml wash + 35 ml elution in a second 250 ml
                          flask.


                      6)  Rotary evaporate the two resulting fractions to about 1 ml.

                      7)  Transfer the 20% fraction (containing dieldrin, endrin,

                                                     6









                          methoxychlor and o,p-DDD) to a centrifuge tube with three hexane
                          rinses. Add O.S al of iscoctane and then N-evap to O.S al. Bring
                          it up to 1 al with isooctane and transf er to a 2 al vial with
                          teflon cap. Rinse the vial previously with acetone, hexane and
                          isoactane. Spike the sample with SO Al of PCB #30 (11.4 ng/ml)
                          before injection into the GC. Take 300 Al into an autosampler
                          vial and load into autosampler.for GC run.

                   C. SILICA GEL  cleanup and fractionation

                       1) Prepare silica gel 60 (70/230 mesh) col--s in the same manner as
                          the florisil column.


                       2) Wash the column with 20 al hexane.


                       3) -When hexane reaches the top of the silica gel, add the 6% florisil
                          eluate (1-2 al) and allow it to drain into the column. Rinse
                          flask with 3 al of hexane and drain into column. Discard eluents.


                       4) Wash the column with 5 al of a 0.5:99.5 ratio of benzene:hexane,
                          followed by 35 al of the solvent. Collect the eluates in a 250 al
                          round-bottom flask. (This is fraction-1, silica gel).

                       5) Elute the columns with 40 al of a 25:75 ratio of diethyl
                          ether:hexane and collect the eluate in a 250 al round-bottom
                          flask. (This is fraction 2, silica gel).


                       6) Rotary evaporate both fractions to about 1 al, then transfer to a
                          centrifuge tube with three rinses of hexane. Add 0.5 al of
                          isooctane and N-evap down to @ 0. 5 al. Bring it up to 1 al with
                          isooctane again and transfer to 2 al vial with teflon cap. Before
                          GC analysis, spike the extract with 50   141 of PCB #30 (11.4 ng/ml),
                          then take 250 Al into an autosampler vial for GC run.


                  D. GAS  CHROMATOGRAPHY determination.


                       1. Silica Gel 25% fraction. Most pesticides come out in this
                          fraction. use autosampler/GC program 9.
                          Program 9 condit@o ns: Injector temperature 230    0C, Detector
                          temperature 350 C. Gas carrier He at 5 psig, makeup gas nitrogen
                          at 45 psig. Equilibrium time 3 min, Total run time 60 min,
                          attenuation S.


                                          Oven temperature program
                                                      .1    2      3     4
                          Oven temp (OC)             120   150   225  280
                          Xso time (min)               3    5     10    15
                          Ramp rate (OC/min)          30    4     20

                       2. Silica gel 0.5% fraction.    PCBs and DDE come out in this fraction.
                          Use autosampler/GC program   6.


                                                       7










                            Program 6 conditions: injector and detector temperatures as well
                            as gas flow rates and everything else remains the same as in
                            program 9, except for the oven temperature program and running
                            time.


                                           Oven temperature program
                                                     1       2     3
                            Oven tamp (OC)          120    260  280
                            ISO time (m1m)           6       0    0
                            Ramp rate (OC/min)       2     20 @

                       3.   Florisil 20% fraction.   Some pesticides come out in this fraction.
                            Use program 9 (see above).


              XIII. DEMONSTRATION OF STATISTICAL CONTROL


                    Statistical control of GC measurements   can be demonstrated graphically by
                    the use of control charts (Taylor 1989, p. 129). Initially, a standard
                    of known concentration will be injected for a total of 7 independent
                    measurements. If the range is linear, the mean relative response factor
                    will be used as the central line to maintain statistical control.
                    Standards will be injected every day that a set of samples is run. If
                    the value of the standard is within 1 standard deviation of the mean"
                    then we can say that we have statistical control. If a known reference
                    standard is used, then the certified concentration value can be used as
                    the central line (Taylor 1989, p. 131). The control limits will be--
                    evaluated by the control charts.


                    In addition, for every set of 10 samples one sample will be    run im
                    triplicate.  The calculated concentrations will be compared.     If the CV
                    (coefficient of variation is 'i 10%, then we can assume that our
                    measurements are within our established method precision. The use of
                    standards of known concentrations will allow to construct standard
                    reference calibration curves against which the sample runs will be
                    compared. If an outlier is suspected, the calculations and data
                    transfers will be rechecked. If the results are still suspect then the
                    sample before and after suspect and the suspect sample will be
                    reanalyzed. A value will be considered an outlier if there is an
                    assignable cause.


              XIV. CALCULATIONS


                    The concentration of PCBs and OCs will be determined using the internal
                    standard method to eliminate injection variability and the need to
                    maintain the sample at a constant final volume.

                    A) Organochlorine pesticides: Pesticides will be quantified based on an
                       internal standard (PCB 30) added to the samples after the extraction
                       step. Quantification is carried out by calculating relative response
                       factors based on peak areas.









                   B) Total PCBs: PCBs will be quantified with the use of CONSTAR (see
                      aOMSTAR SOP).

              XV.  CONFIRMATION AND ASSIGNMENT OF UNCERTAINTY


                   organochlorine pesticides will be confirmed in approximately 10% of the
                   samples by GC/MS. This confirmation may only be possible for compounds
                   detected at significant concentrations.

                   Assignment of uncertainty: A range performance chart will be constructed
                   where the relative response factors (RRFs) at low, middle, and high
                   concentrations will be plotted vs concentration. The upper warning limit
                   (UWL) and lower control limit (LCL) will be the 95% C1, and the upper
                   control limit (UCL) the 99.7% CI. Samples with values above the UCL will
                   be diluted and reanalyzed; those with values below the LCL will be tagged
                   as below detection limit.





















































                                                    9














                                                       TABLE I


                           Retention times and limits of detection of OC pesticides



                      Compound                         Retention       Method           Limit of
                                                       time (=in)      detection        detection
                                                                       limit            (Ug/ml)
                                                                       (ng/ml)



                      HCB                              14.27
                      ga--a-HCH                        14.93           2.3              0.8
                      Int.std PC3 30                   15.41
                      Haptachlor                       19.37           1.1              0.4
                      Aldrin                           21.22           1.9              0.6
                      Keptachlor  epoxide              23.06
                      Oxychlordane                     23.27
                      gamma-chlordane                  24.17
                      o,pI-DDE                         24.61
                      Endosulfan                       24.79
                      p,pI-DDD                         25.04
                      Ct-Chlordane                     25.49
                      Dieldriz                         26.06           1.3              0.4
                      p,pI-DDE                         26.12
                      t-Nonachlor                      26.38
                      Endrin                           26.93           s.6              1.9
                      Endosulfan 11                    27.06
                      o,PI-DDD                         27.90
                      p,pI-DDT                         30.06           12.6             4.2
                      Metboxychlor                     33.71



                        Coll- DB-1, Autosampler method 9; see SOP for GC conditions and
                      procedures.



















                                                         10










































































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