SEPA
The Penobscot  River and  Environmental  Contaminants:
Assessment of Tribal Exposure Through Sustenance Lifeways
U.S. EPA Region I
Regionally Applied  Research  Effort
Final RARE Report
August 2015

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                                       PREFACE
This study demonstrates EPA New England's strong commitment to build partnerships with EPA
Office of Research and Development (ORD), EPA laboratories and toxics experts throughout the
agency and other federal agencies while fully supporting the principles outlined in EPA's Indian
Policy (Ruckelshaus, 1984; reaffirmed in January 09, 2014). On January 9, 2014, Gina McCarthy,
EPA Administrator, issued an All EPA Employee Memorandum reaffirming the Indian Policy, below
is an excerpt from that reaffirmation memorandum.

   "The U.S. Environmental Protection Agency in 1984 became the first federal agency to adopt a formal
   Indian Policy. Today, I am proud to formally reaffirm that policy. By my action, the EPA reiterates its
   recognition that the United States has a unique legal relationship with tribal governments based on the
   Constitution, treaties, statutes,  executive orders and court decisions. The EPA recognizes the right of the
   tribes as sovereign governments to self-determination and acknowledges the federal government's trust
   responsibility to tribes. The EPA works with tribes on a government-to-government basis to protect the
   land, air and water in Indian Country. . . .

   The reaffirmation of the Indian Policy articulates  the importance of our tribal programs and our
   relationship with tribal governments. Our work in Indian Country is crosscutting and affects all aspects
   of the EPA's day-to-day functions. The environmental challenges we face are many. We must protect
   our precious water resources and address chemical safety. And we must continue taking common-sense
   steps to reduce the harmful carbon pollution that fuels climate change. Only through continued
   partnership with tribes can we truly achieve a cleaner, healthier and more prosperous America today
   and for future generations.

   It is  an important time in our partnership with tribes as the EPA builds on past successes and strives to
   meet current and future environmental challenges in Indian Country. Please join me in advancing our
   strong partnership with tribal governments to protect human health and to safeguard the environment in
   Indian Country."
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Final RARE Report August 2015

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                                     Disclaimer
This report was funded wholly or inpart by the United States Environmental Protection Agency (EPA).
This report has been subjected to EPA's peer review process and has been approved for publication as
an EPA document.  Mention of trade names or commercial products does not constitute endorsement or
recommendation for use. All research projects making conclusions or recommendations based on
enviromental data and funded by EPA are required to participate in the Agency's Quality Assurance
Program. This project was conducted under an approved Quality Assurance Project Plan. This report
includes peer-reviewed scientific conclusions about environmental quality; it does not include EPA
policy determinations about whether or how to respond to those conclusions.
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                      FINAL REPORT
        Regional Applied Research Effort (RARE) Project
                          August 2015


    The Penobscot River and Environmental
Contaminants: Assessment of Tribal Exposure
            through Sustenance  Lifeways

                     Investigative Organizations
                  US Environmental Protection Agency
                        US Geological Survey
             Agency for Toxic Substances and Disease Registry
                     US Fish and Wildlife Service
                       Bureau of Indian Affairs
         Penobscot Indian Nation - Department of Natural Resources
                        Report Prepared by
           Valerie Marshall1, Richard Sugatt1, Robert Hillger1, Adria
        Elskus4, Christian Byrne3, Joseph Ferrario3, Carl Orazio4, Robert
           Dudley4, Sarah Warren2, David DeMarini2, Adam Swank2,
              Thomas Hughes2, Janet Diliberto2' Steve Stodola l,
          Steve Mierzykowski5, Katie Pugh6, and Charlie Culbertson4.
                          Acknowledgements
            Dan Kusnierz, Robert Lent, Steve DiMattei, Dave McDonald,
    Alan Van Arsdale, Gary Perlman, Jim Lazorchak, Larry D. Claxton, Linda S. Birnbaum,
           Jason Mitchell, Jan Paul, Joe Dana, Kristin Peet, Michele Attean,
                Jason Sockbeson, Dan Morse and, Leann Jensen.
(1) EPA Region 1                     (4) US Geological Survey
(2) US EPA Office of Research and Development  (5) US Fish & Wildlife Service
(3) US EPA Office of Pesticides Program       (6) Agency for Toxic Substance & Disease Registry
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                                       Abstract
EPA in collaboration with the Penobscot Indian Nation, U.S. Geological Survey (USGS), Agency
for Toxic Substances and Disease Registry (ATSDR), and the U.S. Fish and Wildlife Service
(USF&WS) collectively embarked on a four year research study to evaluate the environmental
health of the riverine system by targeting specific cultural practices and using traditional science to
conduct a preliminary contaminant screening of the flora and fauna of the Penobscot River
ecosystem. This study was designed as a preliminary screening to determine if contaminant
concentrations in fish, eel, snapping turtle, wood ducks, and plants in Regions of the Penobscot
River relevant to where PIN tribal members hunt, fish and gather plants were high enough to be a
health concern. This study was not designed to be a statistically validated assessment of
contaminant differences among study sites or among species.

The traditional methodology for health risk assessment used by the U. S. Environmental Protection
Agency (EPA) is based on the use of exposure assumptions (e.g. exposure duration, food ingestion
rate, body weight, etc.) that represent the entire American population, either as a central tendency
exposure (e.g. average, median) or as a reasonable maximum exposure (e.g. 95% upper confidence
limit).  Unfortunately, EPA lacked exposure information for assessing health risks for New England
regional tribes sustaining a tribal subsistence way of life. As a riverine tribe, the Penobscot culture
and traditions are inextricably tied to the Penobscot River watershed. It is through hunting, fishing,
trapping, gathering and making baskets, pottery, moccasins, birch-bark  canoes and other traditional
practices that the Penobscot culture and people are sustained. The Penobscot River receives a
variety of pollutant discharges leaving the  Penobscot Indian Nation (PIN) questioning the
ecological health and water quality of the river and how this may affect the practices that sustain
their way of life.

The objectives of this Regional Applied Research Effort (RARE)  study were to:
   >  Develop culturally sensitive methodologies for assessing the potential level of exposure to
       contaminants that Penobscot Indian Nation tribal members may have from  maintaining
       tribal sustenance practices.
   >  Conduct field surveys and laboratory analysis on targeted flora and fauna for chemical
       exposure to dioxins/furans, polychlorinated biphenyls (PCBs), total mercury and methyl-
       mercury.
   >  Assist the Agency for Toxic Substances and Disease Registry (ATSDR) by providing the
       necessary data to conduct a Public  Health Assessment for the Penobscot Indian Nation.
   >  Establish protocols for assessing the level of exposure to PCBs,  dioxins/furans and mercury
       to PIN tribal members as a consequence of gathering tribal plants for medicinal and
       nutritional purposes; as well as consuming fish, wood duck, and snapping turtle as  a primary
       source of nutrition.
   >  Survey surface water, drinking water, and sediment from the Penobscot River and Indian
       Island to assess the exposure of PIN tribal members to environmental genotoxicants that
       continue cultural sustenance practices.

This research initiative collected and analyzed  sediment and biota to determine the level of
contaminant exposure to Penobscot tribal members. Natural resource utilization patterns and
exposure pathways were identified based on discussions with the Tribal elders. Identification of
Tribal exposure factors (exposure pathways and contaminant concentrations) was essential for
accurately assessing potential long-term Penobscot Indian Nation  tribal members'  exposure.
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Based on this study, ATSDR's Public Health Assessment (PHA) concluded that the Penobscot Indian
Nation (PIN) tribal members who eat fish and snapping turtle at the ingestion levels suggested in the
Wabanaki Traditional Cultural Lifeways Exposure Scenario Report (Wabanaki Exposure Scenario)
may be exposed to harmful levels of mercury, dioxins/furans, dioxin-like PCBs, and other PCBs.
ATSDR is most concerned about mercury in fish and snapping turtle taken from the Penobscot River.
Mercury is most harmful to children and developing fetuses. It is especially important for pregnant
and breastfeeding women, women who may become pregnant, and children to limit their
consumption offish and snapping turtle in order to decrease their risk of neurological damage due to
mercury exposure.  The ATSDR recommends that Penobscot Indian Nation tribal members follow
the existing Penobscot Indian Nation Department of Natural Resources' fish advisory and the State of
Maine  Safe Eating Guidelines for all fish caught in the Penobscot River and limit their consumption
of snapping turtle. ATSDR recommends that PIN members eat only 1-2 fish meals per month from
the Penobscot River, and limit their consumption of snapping turtle to 2-3 meals per month.  If
Penobscot River fish and turtle are both eaten, ATSDR recommends no more than some combination
of 1-2 (10 oz.) servings offish, or 2-3 (8 oz.) servings of turtle per month.

The EPA preliminary risk  assessment is consistent with ATSDR's PHA recommendations because it
indicates that consumption offish (especially eel) and snapping turtle at the Wabanaki  Exposure Inland
Non-Anadromous tribal consumption rates is associated with a risk of potential concern. (See Exposure
Assessment Section)

ATSDR indicates that PIN tribal members who eat wood duck, fiddlehead fern, or medicinal roots at
the Wabanaki Exposure Scenario-suggested ingestion rates from the areas where the samples were
collected for this study should not be exposed to harmful levels of mercury, PCBs, dioxins/furans or
dioxin-like PCBs. As shown in the Exposure Assessment section, EPA's preliminary risk assessment
is consistent with these ATSDR recommendations.   ATSDR also indicates that incidental ingestion of,
and dermal exposure to, Penobscot River sediment should not pose a human health hazard.

The Salmonella mutagenicity assay was used to assess the mutagenic potencies of organic extracts of
the Penobscot River water and sediment, as well as  of drinking water samples. Mutagenicity is a
statistical indicator of some cancer-causing (carcinogenic) chemicals.  Most samples were either not
mutagenic or, compared to published data for comparable extracts, had low to moderate mutagenic
potencies. Thus, there is little evidence that extracts of these environmental media have mutagenic
activity that might be due to the classes  of compounds that this assay readily detects, such as polycyclic
aromatic hydrocarbons, nitroarenes, and aromatic amines.
Page  v                                                                      201

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                                    Table of Contents
Abstract	v
Executive Summary	1
Background (The Relationship of the Penobscot River to the Penobscot Indian Nation)	5
Description of the Study Area (Penobscot River Watershed)	6
Physical Setting	9
    Study Area	9
    Demographics 	9
    Climate 	9
    Geomorphology	9
    Hydrology 	10
Penobscot River Watershed Issues	11
Health Advisories	12
Penobscot River Point Source Discharges	16
Previous Investigations	18
Purpose and Objectives of Research 	20
Research Responsibilities 	21
  Research Team	22
Approach	24
Research Study Location	25
Sample Collection Design  and Analysis	32
    Research Project Schedule	32
    Sample Collection and Preparation 	32
    Sample Collected	33
      Sediment	35
      Fish 	36
      Plant	38
      Turtle 	39
      Duck	40
      Surface Water, Drinking water, and Sediment for Mutagenicity Analysis	41
Summary of Chemical Extraction Methods	42
    Sediment/Vegetation 	42
    Tissue	42
    Water and Sediment for Mutagenicity Analyses 	42
Sample Analyses	43
    Sediment 	43
    Fish Tissue 	44
    Turtle and Duck Tissue	44
    Plants	44
Analytical Procedures	44
    Total Mercury 	44
    Methyl Mercury	45
    Congener-Specific PCBs 	45
    Dioxin/Furans and co-planar PCBs 	46
    Mutagenicity Assays	46
Quality Assurance	48
Peer Review	49
Data Validation	50
Analytical Results from Reaches 1-6: Contaminant Concentrations	53
    Summary of Contaminant Concentrations 	60
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Exposure Assessment of Penobscot Indian Nation tribal members 	63
       Risk Assessment vs. Risk Management 	63
       EPA Exposure Assessment	63
       Toxicity Assessment	66
       Risk Characterization 	69
       Risk Management Criteria	71
       Preliminary Risk Results	72
       Hazard Quotients	77
       Cancer Risk	78
       Uncertainty Analysis	79
Health and Exposure Assessment Conclusions and Recommendations	80
       ATSDR Health Assessment	80
       ATSDR Health Assessment Recommendations	81
       US EPA Exposure Assessment Recommendations 	81
       Recommendations for further Investigations	82
Conclusions and Recommendations of Mutagenicity Study	83
       Analytical Results of Salmonella Mutagenicity Study	83
       Conclusions of Salmonella Mutagenicity Study	86
       Salmonella Mutagenicity Recommendations	87
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                                           Tables
Table 1: Summary of Composite Samples Collected of Sediment, Flora, and Fauna	34
Table 2: Summary of Samples Collected for Mutagenicity Assays	35
Table 3: Summary of Sample Types Collected for Chemical Analyses 	43
Table 4: Data Validation for Penobscot RARE Data	52
Table 5: Contaminant Concentrations in Sediment and Biota in Reach 1	54
Table 6: Contaminant Concentrations in Sediment and Biota in Reach 2	55
Table 7: Contaminant Concentrations in Sediment and Biota in Reach 3	56
Table 8: Contaminant Concentrations in Sediment and Biota in Reach 4	57
Table 9: Contaminant Concentrations in Sediment and Biota in Reach 5	55
Table 10: Contaminant Concentrations in Sediment and Biota in Reach 6	59
Table 11: Highest Contaminant Concentration in Sediment and Biota by Sample Type	61
Table 12: Highest Contaminant Concentrations in Sediment and Biota by Reach	62
Table 13: Food Categories and Consumption Rates for Different Tribal Populations	64
Table 14: Exposure Assumptions & Toxicity Factors	67
Table 15: Dioxin Toxicity Equivalence Factors	68
Table 16: Risk Based Concentrations	69
Table 17: EPA Acceptable Cancer Risk Management Criteria	71
Table 18: Adult Risk at Inland Non-Anadromous Tribal Ingestions Rate	74
Table 19: Summary of Mutagenicity Samples of Drinking Water	84
Table 20: Summary of Mutagenicity Samples of River Water	55
Table 21: Summary of Mutagenicity Samples of River Sediments 	86

                                           Figures
Figure 1: Penobscot River Basin and Surrounding Watersheds	1
Figure!: Penob scot River Basin and Sub-basins  	8
Figure 3: Major Point sources of the Penob scot River	16
Figure 4: Penobscot River Study Six Reaches	27
Figure 5: Detail of study Reaches  1-2	28
Figure 6: Detail of study Reaches  3-4	29
Figure 7: Detail of study Reaches  5-6	30
Figure 8: Mutagenicity sample collection locations 	31
Figure 9: Chart of Adult Risk by Reach	75
Figure 10: Chart of highest contaminant concentrations in sediment and biota by reach	76
Figure 11: Hazard Quotients of Potential Concern	77
Figure 12:  Hazard Quotient of Potential  Concern per species per Reach	77
Figure 13:  Cancer Risk of Potential Concern for fauna	78
Figure 14:  Cancer Risk of Potential Concern for flora	78
                                         Appendices
A. Abbreviations  	88
B. References Cited 	90
C. Penobscot RARE Project Schedule	93
D. Personnel Associated with the RARE Study	97
E. Quality Assurance Statement	 104
F. Risk of Each Biota Type in each Reach at the Inland Non-Anadromous Tribal Ingestions Rate	 106
G. Contact Information for Peer Review Panel	114
   Page  viii                                           Final RARE Report August 2015

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Penobscot tribal members supplement their diets by fishing on the Penobscot River
                                             Penobscot tribal members harvesting fiddlehead ferns.
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Final RARE Report August 2015

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                               Executive Summary
This study was designed to assess potential exposure to contaminants and the concentrations of
those contaminants along the Penobscot River in areas frequented by Penobscot tribal members
while gathering, hunting and fishing. This is a preliminary screening that determined if
contaminant concentrations in fish, snapping turtle, wood duck, and plants in regions of the
Penobscot River relevant to where PIN tribal members hunt, fish and gather plants were high
enough to be a health concern. Based on the limited funds available, this study was not designed to
be a statistically validated assessment of contaminant differences among study sites or among
species.

As a riverine tribe, the Penobscot culture and traditions are inextricably tied to the Penobscot River
watershed.  It is through hunting, fishing, trapping, gathering and making baskets, pottery,
moccasins, birch-bark canoes and other traditional practices that the Penobscot culture and people
are sustained. If these traditional activities are not continued, the very words of the Penobscot
language that describe these practices will be lost. The ability to preserve the PIN's Native
American culture is being lost. The ecosystems that support the flora and fauna to sustain the
PIN's subsistence way of life are contaminated by toxic pollutants discharged in the air, water, and
land on and near the Tribes' trust and reservation lands. The PIN Tribal Council is very concerned
that engaging in traditional cultural activities is harming tribal members. Some members are not
continuing to sustain a traditional lifestyle due to the fear of the harmful health effects and
depleted resources; especially since the PIN's rates for lung and cervical cancer are some of the
highest in the State of Maine (Valcarcel 1994 and Miller 1994).

The Regional Applied Research Effort (RARE) is an ORD program that provides funding to the
Regions to enter into innovative research partnerships with ORD and address near term research
needs.  RARE projects address a wide array of environmental science issues critical to ORD's regional
partner communities and address regional and national research priorities. The goals of the program
are to:
   •  Provide the regions with near-term research on high-priority, region-specific science needs;
   •  Improve collaboration between regions and ORD laboratories and centers; and,
   •  Build a foundation for future scientific interaction between ORD and the regions.

 This RARE study was designed to provide the scientific information needed to link the science to
 policy and regulatory decision-making within Indian Country. This RARE project is a significant
 multi-Agency study that was designed in a collaborative initiative with 10 partners, i.e., US EPA
 [Region 1 Boston and Chelmsford, Office of Pesticide Programs (OPP), Office of Research and
 Development (ORD)], and the National Center for Environmental Assessment (NCEA),
 USF&WS, ATSDR, USGS [Maine Water Science Center and the Columbia Environmental
 Research Center (CERC)], and PIN. The purpose of the RARE study was to identify unique
 exposure pathways so that scientifically sound data could be collected  and culturally sensitive
 methodologies could be developed for assessing the level of harm the PIN may be exposed to by
 maintaining tribal sustenance practices.

 This research initiative examined the potential adverse risks of exposure of the PIN tribal members to
 contaminants in sediment and biota through ingestion. The approach for this research project combined
 some  of the elements of consumption surveys such as interviewing Tribal elders to determine recent
 natural resource utilization patterns with careful identification of Tribal exposure factors (contaminant
 concentrations, pathways of exposure). Collaborating with numerous scientists and ATSDR assured
 the  scope and procedures identified  for this project met the objectives of the  PIN  and that  the

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 methodologies employed were accepted within the scientific community. To ensure the highest quality
 and reliability, a process of internal and external peer review by both cultural and scientific experts was
 followed.

 The process used was culturally sensitive, respectful, drew on traditional environmental  knowledge
 (such as the observational expertise of elders), and was developed in partnership with tribal cultural
 and technical experts.

The data collected for this preliminary screening can assist the PIN when developing health
advisories, making decisions regarding PIN's tribal members' health, and the PIN's environmental
resources. This study enables the PIN to make decisions based on tribal consumption and exposure
rates rather than relying on rates developed for the average American, which is the standard
typically applied by EPA regulators. Since EPA has been working with Tribal Nations to develop
models for Native American culturally based exposure risk assessments, this study may be
transferable to Tribal Nations across the country.  Both the exposure pathways identified and the
detection limits of the analytical methods used  for this  study are transferable to Tribal Nations that
have diets similar to those studied in this report.

Study Design
The research approach for this study was comprised of two parts: identifying the flora and fauna
used by the PIN in sustaining their traditional way of life, and selecting specific geographical
locations (reaches) along the Penobscot River to collect flora, fauna, surface water and  sediment.
Six reaches deemed ecologically representative were selected along 87 miles (140 kilometers) of
the Penobscot River between Old Town and Medway, Maine. The reaches were chosen based on
previous sediment mapping conducted by the USGS in 1999 (Dudley and Giffen, 2001). The  1999
mapping effort involved the use of ground-penetrating  radar data to characterize the bed-sediment
composition in selected reaches of the Penobscot River. Sampling locations were chosen on the
basis of the mapping information and other river characteristics including wading and swimming
areas, depositonal zones within the channel, and sites upstream and downstream of river features
that control or potentially impact sediment transport (such as dam structures and impoundments).
The reference (control) reach included both free flowing (East Branch Penobscot) and natural lake
waters (Salmon Stream Lake) that were within  the upper Penobscot watershed and upstream of
any known pollution point sources other than regional air deposition.

Field sampling of sediment, fish, duck, turtle and plants occurred from May 2008 to October 2009.
All sampling procedures followed EPA-approved protocols as outlined in the Quality Assurance
Project Plan  (QAPP) for this project (Orazio, 2008). The EPA ORD also conducted several  audits
of the field work to ensure compliance with the project QAPP. The river locations sampled
represent a variety of conditions, ranging from  relatively undisturbed, undeveloped conditions
(e.g., East Branch of the Penobscot River) to more developed conditions (e.g., the dam  impounded
area north of Old Town).

Shallow-water sediment composites consisting of two to five grabs were collected  at each of the
six reaches. Sediments were collected from areas where PIN tribal members typically wade in the
water when hunting, fishing and gathering medicinal plants.

Small-mouth bass (Micropterus dolomieu\ chain pickerel (Esox niger), white perch (Morone
americana\  yellow perch (Percaflavescens), brown bullhead (Ameiurus nebulosus) and American
eel (Anguilla rostrata) were collected from each of six reaches. Fish were collected by boat
electro-shocking, gill netting, trap netting and hook and line. For each reach, one composite

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sample was prepared of each species (a "species composite") consisting of three to five fish of
similar size of each species.

A total of seven composite fiddlehead fern samples were obtained, representing five of the six
reaches and one field duplicate sample. A total of five composite medicinal plant samples were
collected representing four reaches and one duplicate.

 Snapping turtles (Chelydra serpentina) are an important tribal food. Snapping turtles are a long-
 lived, upper trophic level aquatic species that readily accumulate contaminants.  Seven snapping
 turtles from five of the six reaches were collected for the study.

 Wood duck (Aix sponsa) were collected and analyzed because wood duck are currently the most
 hunted duck species by PIN tribal members. Since wood duck are a common breeder on the
 Penobscot River, they are more likely to reflect local contaminant levels than other waterfowl that
 use the river more seasonally. Sixteen wood ducks were collected, representing five composite
 samples from four of the six river reaches.

 Samples collected for chemical analysis in this study were taken from the biota identified above
 and river sediment. Labs analyzed samples for dioxins/furans; PCB congeners (including dioxin-
 like PCB congeners); methyl mercury (Me Hg); and, total mercury (Hg). Labs provided 2,3,7,8-
 TCDD toxic equivalent (TEQ) values for dioxins/furans and dioxin-like PCBs.

 Samples collected for mutagenicity analysis in this study included river water, drinking water,
 and river sediment. River water samples were taken at three locations: (1) at an upstream control
 headwater location; (2) immediately below the effluent discharge of a municipal and/or industrial
 site, and; (3) a downstream (dam) site. Drinking-water samples were taken from a convenient tap
 at the PIN laboratory.  River sediment samples were taken at approximately the same sites as the
 river water samples and at Indian Island.

 For this report, EPA considered cancer risks of 1E-06 or less and non-cancer Hazard Quotients of one
 or less to be of "no concern" and risks greater than these levels to be of "potential concern".  These
 risk management criteria were selected because they are consistent with a variety of EPA regulatory
 programs.

 Findings
 With the flora and fauna data collected from this study, ATSDR conducted a Public Health
 Assessment (PHA) for the Penobscot Indian Nation and EPA conducted a preliminary risk
 assessment. EPA compared the concentrations in biota to risk-based concentrations to determine
 the level of risk to the Penobscot Indian Nation tribal members that maintain cultural practices
 and sustenance lifeways associated with the Penobscot River. EPA's risk results are consistent
 with ATSDR's consumption recommendations. EPA risk results are based upon a preliminary
 risk assessment that can be  found in the Exposure Assessment Section of this report.  The results
 from EPA's preliminary risk assessment suggest that the consumption of each animal species
 except duck at the Wabanaki Exposure Scenario consumption rates is associated with a risk of
 potential concern.  Therefore, ATSDR's PHA recommendations limiting fish consumption to 1-2
 meals per month from the Penobscot River and snapping turtle consumption to 2-3 meals per
 month are not inconsistent with EPA's preliminary risk assessment. EPA also concurs with
 ATSDR's conclusion that mercury was not found at levels of health concern in wood duck,
 fiddlehead fern, or medicinal plants.


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ATSDR concluded that the Penobscot Indian Nation (PIN) tribal members who eat fish and snapping
turtle at the ingestion levels suggested in the Wabanaki Traditional Cultural Lifeways Exposure
Scenario Report (Wabanaki Exposure Scenario) may be exposed to harmful levels of mercury,
dioxins/furans, dioxin-like PCBs, and other PCBs.

ATSDR is most concerned about mercury in fish and snapping turtle taken from the Penobscot
River. Mercury is most harmful to children and developing fetuses. Therefore, it is especially
important for pregnant and breastfeeding women, women who may become pregnant, and children
to limit their consumption offish and snapping turtle in order to decrease their risk of neurological
damage due to mercury exposure. Tribal members should follow the existing Penobscot Indian
Nation Department of Natural Resources' fish advisory and the State of Maine Safe Eating
Guidelines for all fish caught in the Penobscot River. To be safe, ATSDR recommended that PIN
members eat only 1-2 fish meals per month from the Penobscot River and limit their consumption
of snapping turtle to 2-3 meals per month.  If Penobscot River fish and turtle are both eaten,
ATSDR recommended no more than some combination of 1-2 (10 oz.) servings offish, or 2-3 (8
oz.) servings of turtle per month.

ATSDR concluded that PIN tribal members who eat wood duck, fiddlehead ferns,  or the medicinal
plants similar to the ones tested for this study and in the same locations where the samples for this
study were taken will not be exposed to harmful levels of mercury, PCBs, dioxins/furans or dioxin-
like PCBs.  ATSDR also found that incidental ingestion of, and dermal exposure to, Penobscot River
sediment in the same locations where the samples for this study were taken do not pose a human
health hazard.

The findings from the mutageniciry testing showed that most of the collected samples  were not mutagenic
or had a low to moderate response. Mutageniciry is a feature of some cancer-causing (carcinogenic)
chemicals. The main conclusions were that the drinking water, Penobscot River water, and Penobscot
River sediments exhibited little mutagenicity. Based on these results, there is not a concern for the
presence of mutagenic compounds that this assay detects, e.g., polycyclic aromatic hydrocarbons (PAHs),
nitro-PAHs, and dyes, in any of the samples tested.

Recommendations
The preliminary study  indicates that contaminant concentrations are high enough to warrant further
investigation of both human health and ecological risk. Therefore, EPA recommends additional study
to statistically characterize how contaminants are related to type, size, and location offish to support
risk-based recommendations to PIN members concerning consumption offish at different river
locations. EPA also recommends that the unused,  frozen fish tissues (offal) be analyzed to estimate
the contaminant concentrations in whole fish that would be consumed by fish-eating wildlife (e.g.
mink, eagle, snapping turtle) to evaluate ecological risk to higher trophic level predators. The
additional data could also serve as a baseline for tracking changes in contaminant concentrations over
time.  Any further  studies should be coordinated with the PIN Health Department in their effort to
correlate the health results with fish consumption and track changes in fish consumption behavior
through education and  issuance of health advisories to PIN members.
Page  4                                                                       201

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                                      Background
         The Relationship of the Penobscot River to the Penobscot Indian Nation
       ^nADKB0^XA!^nx>$? »'
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                                                                                             -
Mount Katahdin and the Penobscot River
 Generating a research study of this nature requires field scientists to understand the intricate connections
 between the ecology of a riverine system and the people that sustain life from this ecosystem. The
 Penobscot River is of great importance to the Penobscot people and has been the center of the Tribe's
 existence for thousands of years. The Penobscot Indian Nation dates back approximately 9,500 years.
 Important burial and ceremonial sites are located upon these islands, which are generally forested and
 low-lying, with extensive floodplains and forested wetlands. Traditional activities take place on and
 around the islands including hunting, fishing, trapping, gathering, boating, camping, sweat lodges and
 ceremonies.  The floodplains support an annual household and commercial harvest of fiddlehead ferns.
 Indian Island, near Old Town, Maine, is the primary residence and the seat of tribal government for the
 PIN.

Penobscot Indian Nation sustenance fishing rights were reserved through historical treaties with
Massachusetts and Maine, and the 1980 Maine Indian Land Claims  Settlement Act. The PIN acquired its
status as a federally recognized Indian Nation in 1980 from the BIA. As part of the statutory provisions
granting recognition, the PIN is entitled to protect and preserve the natural resources of its recognized
trust and reservation lands. These regulations provide that the PIN shall have exclusive authority to
promulgate and enact ordinances regulating hunting, trapping, fishing or other taking of wildlife within
their respective Indian territory.

However, fish contamination prevents this right from being fully exercised and may seriously threaten
the health of community members and their traditional lifeways. The ecosystems that support the flora
and fauna historically used by the PIN are contaminated by air, water, and land pollution so that many of
these traditional activities  cannot be carried  out without fear of harmful health effects. PIN tribal
members are fearful of carrying out their traditional practices such as gathering medicinal plants from the
Penobscot River.  They fear eating natural foods such as turtle and duck meat. They fear using raccoon
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fat to make their birch bark canoes.  If these traditional activities are not continued, the very words of the
Penobscot language that are used to describe these practices will be lost.  If tribal members lose the
ability to make baskets, pottery, moccasins, birch-bark canoes, gather medicinal plants, engage in
traditional fishing, and much more, then they lose the ability to preserve their Native American culture
by preserving and passing along traditional lifeways to future generations.
   Basket making by the Penobscot tribal members is a revered cultural practice as well as a source of economic
   income. The reeds used for basket making come from the Penobscot River ecosystem.

                           Description of the Study Area
Penobscot River Watershed: The Penobscot Pviver Basin is centrally located within the borders of
Maine. The Penobscot Watershed is one of the largest watersheds in Maine. Many of the waterways in
this Basin retain their Penobscot names. It is home to the Penobscot people that live on Indian Island,
located in the southern portion of Penobscot River (See Figures 1-2). Because Indian Island is located in
the downstream portion of the watershed, the PIN is potentially affected by the cumulative impacts of the
many point and non-point sources of pollution to the River.

The Penobscot River /pa'nrjbskat/ is New England's second largest river system.  The Penobscot River
drains approximately one-quarter of the State and has a drainage area of 8,588 square miles (22,243 km2)
at its mouth (Fontaine, 1981). Its West Branch rises near Penobscot Lake on the Maine/Quebec border;
the East Branch Pond near the headwaters of the Allagash River (See Figure 1-2). The main stem is 264
miles in length and empties into Penobscot Bay near the town of Bucksport. The landscape of the
watershed includes Maine's highest peak, Mt. Katahdin, rolling hills and extensive bogs, marshes and
wooded swamps. There is a rich history of cultural, social, and economic tradition associated with the
Penobscot River. The Penobscot River is best known for its large historic salmon run (50,000 or more
adults) and its much smaller contemporary run, which is the largest Atlantic salmon run remaining in the
United States (1,000-4,000 adults in recent decades).
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Final RARE Report August 2015

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        Penobscot Basin
  and Surrounding Watersheds
        Androscoggin  -\f '

-------
          ..>

     f     U
          h-
West Branch
 Penobscot

East Branch f
JPenobscot £

                                                          y \Mattavvamkeag
                                                      \
    Pcnobscot Basin
     and Subbasins
                 i   Penohscol
       Pollution Sources: EPA
         (NPDES Major Permits)
       Penobscot Ind. Reservation Islands
       Rivers: NHD
       Subbasins NRCS HUC
       Elevations. USGS SRTM
       EPA Region 1 Map Tracker ID: 10012
Figure 2: Penobscot River Basin and Sub-Basins
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                                        Final RARE Report August 2015

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Physical Setting
Study Area
The study area is located within the Penobscot River Watershed as depicted in Figures 1-2 above. The
study area includes selected reaches along 87 miles (140 km) of the Penobscot River between Old Town
and Medway, Maine. The six sampling areas for the study were chosen to include three of the same sites
surveyed by USGS in 1995-96 and 1999, as well as new sites where there were known or potential areas
of fine-grained sediment deposition (See Figures 4-5). The selection of reaches for this study was further
limited by the proximity of motor-boat launch sites to river reaches deep enough for navigation.

Demographics
The combined population of the towns along the Penobscot River in the study area is approximately
26,000 people. About 40 percent of this population (11,200) is distributed at the downstream limit in the
towns of Old Town and Milford and in part of the Penobscot Indian Nation Reservation.  The Penobscot
Indian Nation Reservation includes the islands and surrounding waters upstream from the Milford Dam -
totaling more than 200 islands. At present, the only permanent settlement, as well as the seat of
government of the Penobscot Indian Nation, is on Indian Island at Old Town. According to the PIN, the
current number of Penobscot tribal members is 2,397. The current total population of Indian Island is
606, of which 455 are tribal members.  The towns of Lincoln and Chester have the next largest
population group with a combined population of 6,300.  The remaining population is distributed among
small towns, farms, and sprawling suburban developments.

Climate
The climate in the Penobscot River Basin is typically characterized by mild summers and cold winters.
The average annual temperature is 41°F (5 °C) at a National  Weather  Service (NWS) station in
Millinocket, about 10 miles west of Medway, and 43 °F  (6 °C) at a NWS station near Old Town. Mean
monthly temperatures range from 13°F (-10 °C) in January to 68°F (20 °C) in July at Millinocket and
17°F (-8 °C) in January to 67°F (19 °C) in July near Old Town. The average annual precipitation in the
basin is about 40 inches (101  cm) and is evenly  distributed throughout the year (U.S. National Oceanic
and Atmospheric Administration, 1995 and 1998).

Geomorphology
The Penobscot River valley can be separated into four distinct geomorphic units. From the headwaters of
the East and West Branches of the Penobscot River downstream to the town of Medway is a mountainous
upland area. This area is characterized by high-relief topography which results in high-energy stretches
of the river that are popular with white-water rafters and kayakers. This mountainous terrain, which is
characteristic of the New England central highlands (Denny, 1982), has many ponds and tributary
streams.  Many of these ponds and streams  are or have been  affected by dams for the generation of
hydroelectric power and flow control for log driving (Kelley and others,  1988).  The high-energy white-
water characterization is not true of the dam impoundments in this region. Water movement in the
impoundments is significantly slower than in the high-energy reaches of the river, enabling fine-grained
sediments to settle out and accumulate on the bottom.

The second section of river, running through the New England coastal lowlands (Denny, 1982) from
Medway to Old Town has a broad floodplain and a wider channel than the upstream section. This
section of the river is characterized by numerous low-profile deposit onal islands and sand bars. Bedrock
outcrops and rapids are rare.  Aerial photographs of this part of the river show historical meandering and
braiding of the river channel and indicate formation of islands by erosion and deposition (Kelley et al.,
1988). The West Enfield Dam is located about midway on this second river reach.
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The third section of the Penobscot River, from Old Town to Bangor, is characterized by numerous rapids
and common bedrock outcrops. The Milford Dam is located at the beginning of this river reach. Bluffs
of unconsolidated material dominate the riverbanks, and raised terraces are well developed in several
locations (Kelley et al., 1988).

The fourth section of the river, below Bangor, is tidally influenced and passes through a geomorphic area
classified as the New Brunswick highlands (Denny, 1982). This part of the river is characterized by
bluffs of unconsolidated material and bedrock cliffs, with fringing salt marshes in protected areas (Kelley
etal., 1988).

Hydrology
The Penobscot River originates as two main branches, the East Branch and West Branch. The drainage
divide at the headwaters of the West Branch constitutes the Maine-Canadian border (Figure 1). At the
confluence of the two branches at Medway (Figure 2), the East Branch has a drainage area of
approximately 1,200 square miles (2,900 km2) and the West Branch drains approximately 2,130 square
miles (5,517 km2). From Medway, the Penobscot River flows south for approximately 112 miles (180
km) to the Gulf of Maine where it discharges into the Atlantic Ocean. The Penobscot River drains about
one-quarter of the State of Maine and has a drainage area of 8,588 square miles (22,243 km2) at its mouth
(Fontaine, 1981).  Streamflow in the Penobscot River Basin vary seasonally with high flows typically in
early spring and late fall and low flows generally in the summer  and early fall.

The Mattaseunk Dam, originally built in 1937-40 in the town of Mattawamkeag, is a run-of-the-river
hydroelectric facility producing 19.2 megawatt (MW) of electrical power (Dana Murch, Maine
Department of Environmental Protection, Bureau of Land and Water Quality, oral communication,
2000). The drainage area of the Penobscot River above the dam is 3,355 square miles (8689 km2;
Fontaine, 1981). The Mattaseunk Dam impoundment has a surface area of 1,685 acres and a gross
storage of approximately 915 million ft3 (Dana Murch, oral communication 2000).

The West Enfield Dam was originally built in 1894. In 1986, this dam was replaced by another dam
constructed immediately downstream from the 1894 structure. The West Enfield Dam is a run-of-the-
river hydroelectric facility producing 13MW of electrical power  (Dana Murch, oral communication,
2000). The drainage area of the Penobscot River above the dam is 5,217 square miles (Fontaine, 1981).
The West Enfield Dam Impoundment has a surface area of 1,125 acres and a gross storage of
approximately 490 million ft3 (Dana Murch, oral communication, 2000).  The Piscataquis River joins the
Penobscot River about 1 mile downstream from the dam and drains 1,453 square miles (Fontaine,  1981).

The study reach in the towns of Old Town and Milford includes  areas near Olson Island in the Milford
Dam Impoundment. The Milford Dam, originally built in 1905-06 in the town of Milford, is a run-of-
the-river hydroelectric facility licensed to produce 8 MW of electrical power (Dana Murch, oral
communication, 2000). The drainage area of the Penobscot River upstream from the dam is 7,325 square
miles (Fontaine, 1981). The Milford Dam Impoundment has a surface area of 235 acres and a gross
storage of approximately 98 million ft3 (Dana Murch, oral communication, 2000).
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Penobscot River, Milford Dam downstream of Indian Island
                    Penobscot River  Watershed-wide Issues
  There are approximately 116 dams in place in the Penobscot River basin, 14 of which are major
  hydropower projects that have generally inadequate fish passage. Public and private facilities
  discharge 150 million gallons of wastewater/day to the river, which is equivalent to -2% of the river's
  average daily outflow. Five major NPDES licensed outfalls discharge into the Penobscot River and
  affect the study area (Figure 3). Some known constituents being discharged include suspended solids,
  heat, oxygen-depleting substances, chlorinated  organics, chromium, copper, dioxin, lead,  mercury,
  phenols, vanadium and zinc.  State fish consumption advisories for mercury, PCBs, and dioxins are in
  place for the Penobscot River (See http://www.maine.gov/dhhs/mecdc/environmental-
  health/eohp/fish/2kfca.htm).  Thermal loading throughout the entire watershed affects the
  migration/resting behaviors of mature adult salmon during spawning.

  West Branch:  This vast area occupies 25% of the land in the entire basin.  The Penobscot name for
  the West Branch is Kettetegwewick, meaning "the main branch." This is the canoe route to Katahdin,
  the highest mountain in Maine and the Tribe's most sacred place. Drainage in the West Branch is
  heavily manipulated for hydropower generation. It contains the  largest privately owned hydroelectric
  complex in the country, and it receives wastewater from two pulp and paper mills and two
  municipalities.  Portions are listed as not attaining water quality  standards (WQS) for bacteria, aquatic
  invertebrate communities, and dissolved oxygen.  PIN data indicate significant algal/cyanobacteria
  blooms that originate here and affect the entire  main stem of the Penobscot downriver (-75 miles or
  120 kilometers).
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  East Branch:  This remote area occupies 13% of the land in the entire basin and is extremely
  important to the restoration of self-sustaining populations of Atlantic salmon (Salmo salar). The
  Penobscot name for this section of the river is Wassategwewick, perhaps meaning "place where torches
  used to fish" or "place where light first touches." This is an important fishing area for the Tribe. The
  watershed, including pristine Grand Lake Mattagamon, is threatened by non-point source pollution and
  air deposition from within and beyond the region.  Sources include forestry activities (e.g. timber
  harvesting and  associated roads) camps and other development.

  Mattawamkeag: This tributary is named for the gravel bar that marks the river's confluence with the
  main stem of the Penobscot. The area occupies 17% of the land in the entire basin. The lower-most
  segment of this reach does not attain WQS for bacteria due to untreated wastes.

  Piscataquis: This area occupies another 17% of the land in the entire basin. This "little branch stream"
  was an extremely important Penobscot travel route and contains significant Atlantic salmon-spawning
  habitat.  This drainage is affected by discharges from two large municipal treatment plants, a textile
  mill, non-point source pollution from agricultural and forestry operations, and at least six dams.
  Significant turbidity and suspended solids within the river are caused by erosion in or near poorly
  constructed haul roads, skid trails, and stream crossings. More than 12 livestock farms have no manure
  storage facilities and more than 2,000 acres of cropland adjacent to surface waters are highly erodible.
  Excessive macrophyte and algae growth downstream of one treatment plant suggest the need for
  additional controls at the facility.

  Lower Penobscot: This area occupies 28% of the land in the entire basin and bears the rock drops
  (now dammed) that are the basis for the name of the river and the Tribe.  Two pulp and paper mills
  discharging here result in fish consumption advisories for dioxins, furans and PCBs.  Several segments
  do not attain bacteria standards due to untreated residential and municipal combined sewage overflow
  wastes.  Several tributaries do not attain Water Quality Standards for dissolved oxygen because of
  agricultural  activities. With point sources accounting for 94% of the total phosphorus loads entering
  the river, observed and model-predicted results indicate that -51 Class B river miles will not meet their
  dissolved oxygen standards. HoltraChem,  a chlor-alkali plant closed in 2000, was located on the lower
  Penobscot.  Sediments downstream contain the highest concentrations of mercury in Maine and
  possibly the country.  The plant was licensed to discharge up to 5 pounds of mercury/year directly to
  the Penobscot River and hundreds of pounds/year to the air.  The Lower Penobscot area is the location
  of Indian Island, the home of the PIN.
                                   Health Advisories
In 1987, the State of Maine issued health advisories limiting the consumption offish from the Penobscot
River.  This advisory was for dioxin discharges specific to pulp and paper mills that discharge industrial
waste directly into the Penobscot Indian Nation's reservation.

In 1997, the State of Maine revised the fish advisories in the Penobscot River to include Polychlorinated
biphenyls (PCBs) and mercury. PIN's Natural Resources and Health departments began issuing Tribal
specific health advisories for the Penobscot River in 1998. PIN - DNR - Fish Consumption Advisory
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The PIN DNR and the State of Maine Health Advisories both recommend the following:
    Pregnant and nursing women, women who may get pregnant, and children under age 8 SHOULD
    NOT EAT any freshwater fish from Maine's inland waters. Except, for brook trout and landlocked
    salmon, 1 meal per month is safe.

    All other adults and children older than 8 CAN EAT 2 freshwater fish meals per month. For brook
    trout and landlocked salmon, the limit is 1 meal per week.

As stated in the State of Maine health advisory:
     "It's hard to believe that fish that looks, smells, and tastes fine may not be safe to eat. But the truth
    is that fish in Maine lakes, ponds, and rivers have mercury in them. Other states have this problem
    too. Mercury in the air settles into the waters. It then builds up in fish. For this reason, older fish
    have higher levels of mercury than younger fish. Fish (like pickerel and bass) that eat other fish have
    the highest mercury levels.

    Small amounts of mercury can harm a brain starting to form or grow. That is why unborn and
    nursing babies, and young children are most at risk. Too much mercury can affect behavior and
    learning. Mercury can harm older children and adults, but it takes larger amounts. It may cause
    numbness in hands and feet or changes in  vision. The Safe Eating Guidelines identify limits to
    protect everyone.

    Warning: Some Maine waters are polluted, requiring additional limits to eating fish.

    Fish caught in some Maine waters have high levels of PCBs, Dioxins or DDT in them. These
    chemicals can cause cancer and other health effects. The Maine Center for Disease Control and
    Prevention recommends additional fish consumption limits on the Penobscot River below Lincoln to
    1-2 fish meals a month." Maine CDC Freshwater Fish Safe Eating Guidelines

The Maine DEP web site explains that mercury is a heavy metal that is used in the manufacture of many
consumer goods and is found naturally in small amounts in oceans, rocks, and  soils. Large amounts of
mercury also become  airborne through manmade processes such as burning coal, oil, wood, or natural
gas as fuel, incinerating mercury-containing garbage, and through industrial production processes that
utilize mercury.  Once in the air, mercury can fall to the ground with rain and snow, contaminating soils
and water bodies.

Once mercury is released into the environment it can change to methyl mercury, a highly toxic
compound.  Methyl mercury is easily taken up in living tissue and bioaccumulates (builds up) over time,
causing serious health effects such as neurological and reproductive disorders in humans and wildlife.
Since mercury does not break down in the environment, it has become a significant health threat to
humans and wildlife.  Mercury levels in Maine fish, loons, and eagles are among the highest in North
America. This has led the Maine Bureau of Health to issue a statewide advisory recommending that
pregnant women, women of childbearing age, and young children limit their fish consumption based on
the type offish they consume. The advisories have been in place since 1994 and remain in effect today
because mercury levels in fish have not decreased. See Maine Department of Environmental Protection
web site http://www.maine.gov/dep/mercury/

PCBs are mixtures of up to 209 individual chlorinated compounds (known as congeners), differing in
number and positions of chlorine atoms. There are no known natural sources of PCBs.  Bulk
formulations of PCBs are either oily liquids or solids that are colorless to light yellow. PCBs are
semivolatile chemicals and can exist as a vapor in air. Trace levels of PCBs have no known smell or

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taste.  Many commercial PCB mixtures are known in the U.S. by the trade name Aroclor. PCBs do not
readily break down in the environment and thus may remain there for very long periods of time.
Although PCB production was banned in the late 1970s, approximately 30 to 70% of what was ever
produced is still in use or in the environment (See Advances in Modern Environmental Toxicology,
Volume XXI, Princeton Scientific Publishing Co., Princeton, NJ 1992). PCBs can travel long distances
in the air and be deposited in areas far away from where they were released. PCBs are relatively
insoluble in water; however, a small amount of PCBs may remain dissolved. In an aquatic system such
as a river, most PCBs stick to organic particles and bottom sediments.  PCBs also bind strongly to soil.
Fish accumulate PCBs from the water column, from sediment where they lay their eggs, and from
consuming other prey in the food web of the river system. Terrestrial animals that eat PCB  contaminated
aquatic organisms accumulate PCBs. PCB bioaccumulation can be a chronic issue in long-lived animals
such as turtles, reaching levels that may be many thousands of times higher than in water.
Bioaccumulation in fatty tissue and biomagnification up the food chain results in the highest
concentrations being found in top predator species.

Polychlorinated dibenzo-^-dioxins (PCDD) are a family of 75 different compounds and polychlorinated
dibenzofurans (PCDF) are a family of 135 different compounds that have various levels  of biological
activity. Dioxins/furans are divided into eight groups based on the number of chlorine atoms, which are
attached to the dioxin/furan molecule at any one of eight positions. The name of each dioxin or furan
indicates both the number and the positions of the chlorine atoms. For example, the dioxin with four
chlorine atoms at positions 2,3,7, and 8 on the molecule is called 2,3,7,8- tetrachlorodibenzo-p-dioxin
(2,3,7,8- TCDD, or TCDD), which is the most toxic of the dioxins to mammals and has received the most
attention (ATSDR,  1998). Only those congeners having chlorine substitutions in at least the four lateral
(2, 3, 7, 8) positions have toxic effects mediated through binding to the arylhydrocarbon receptor protein.
Similarly,  certain PCBs lacking chlorine substitution in the ortho-position and some of their mono- and
di-ortho chlorine analogs are isostereomers of 2,3,7,8,-TCDD and have a common mode of action to that
of the dioxins.  This similarity is the basis for their common measure of toxicity, namely, toxic
equivalents (TEQ).  Twelve PCB congeners fall into a category of "dioxin-like" PCBs. Because of their
structure and mechanism of action, they exhibit toxicity similar to that of chlorinated dibenzo-p-dioxins.
However, their toxicities are 0.00001 to 0.1 times lower than the most toxic dioxin, 2, 3, 7, 8-
tetrachlorodibenzo-p-dioxin (TCDD). A toxic equivalency factor (TEF) approach to evaluating health
hazards has been developed and used to some extent to guide public health decisions (see EPA, 1996 and
ATSDR, 2000 for more details).  In short, the TEF approach compares the relative potency of individual
congeners with that of TCDD, the best-studied member of the dioxin chemical class. The concentration
or dose of each dioxin-like congener is multiplied by its TEF to arrive at a toxic equivalent (TEQ), and
the TEQs are added to give the total toxic equivalency. The total toxic equivalency is then compared to
reference exposure levels for TCDD expected to be without significant risk for producing health hazards.
PCDD/PCDFs may be formed during the chlorine bleaching process at pulp and paper mills.
PCDD/PCDFs are also formed during chlorination by waste water treatment plants. They can occur as
contaminants in the manufacture of certain organic chemicals. PCDD/PCDFs are released into the air  in
emissions from municipal solid waste and industrial incinerators and from any poorly  controlled
combustion process, such as backyard trash burning.   Dioxin has often been identified as one of the
most potent human carcinogens, which is supported by the fact that the most potent dioxin (2,3,7,8-
TCDD) has a cancer slope factor higher than any other chemical on the list of approximately 770
chemicals in the EPA Regional Screening Level table.

When released into the air, some PCDD/PCDFs may be transported long distances, even around the
globe.  When released in waste waters, under certain conditions a small amount of the PCDD/PCDFs
congeners may be broken down by sunlight, a fraction of some may evaporate to air, but most attach to

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soil or attach to bottom sediment.  Similar to the PCBs, PCDD/PCDF concentrations typically increase as
they biomagnify up in the food chain, with higher trophic level organisms containing higher levels than
present in lower level prey, and much higher levels than present in the water.
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                        Penobscot River Point Source Discharges

      ME0100803 MILLINOCKET
      WASTEWATER TREATMENT FACLTY
Millin^jcet
                   ME0000167 KATAHDIN PAPER
                                  East Millnockct
                                         ME0000175 KATAHDIN PAPER
                                                                          2A
                                      i
                                               ME0101796 LINCOLN WWTF
                                            cmcam
                                             ME0002003
                                             LINCOLN PAPER
                                             AND TISSUE
                                    ME0100471 OLD TOWN WWTF
                               ME0002020 RED SHIELD ACQUISITION LLC
                                                                                      Drew Plantation
                                                                                 Webster
                                                                                 Plantation
                                                                                     fTO
                                                                      Penobscot River:
                                                                      Pollution Sources
                                                                               10   15 Kilometers
                                                                             5      10 M iles

                                                                      Pollution Sources: EPA
                                                                      (NPDES Major Permits)
                                                                      Penobscot Indian Reservation Lands: EPA
                                                                      Rivers: NHD
                                                                      Base map © 2009 Microsoft
                                                                      Corp and its data suppliers
                                                                          GISmap#8525_3/6/12
                                                                      Updated 1/5/15
                                                                               Auqus
Figure 3: Point sources of pollution to the Penobscot River. Sources are labeled by orange pentagons. Inset
shows location of this area in the Penobscot watershed.
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                                        Final RARE Report August 2015

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Pulp mill, Lincoln, 1915 Contributed by Lincoln Historical Society, MMN Item 31548
http://www.mainememory.net
 There are numerous point sources of pollution to the Penobscot River (Figure 3). The principal
 industries in the Penobscot River Basin are paper manufacturing, sawmills, lumber preservation, and
 other wood products manufacturing. Other industries in the basin include leather and allied product
 manufacturing and textile production (U.S. Bureau of the Census, 2000). The Lincoln Pulp & Paper Mill
 (LP&P; Figure 3, labeled as Lincoln Paper & Tissue) is of particular concern because pursuant to an
 analysis by the Department of the Interior (DOI), the Penobscot Indian Nation's Reservation abuts the
 facility along the bank of the Penobscot River, and one of the facility's wastewater discharge pipes,
 outfall number 1, discharges into the waters of the Penobscot Indian Nation's Reservation. On-site
 industrial operations for the Lincoln Pulp & Paper Mill began approximately in 1827. The 385-acre
 parcel is currently owned and operated by Lincoln Paper and Tissue (LP&T). This industrial site has
 been used as a grist mill, a saw mill, and wood pulp and paper manufacturing.

 Between June 1983 and August 1998, LP&P was responsible for 276 releases of hazardous,
 nonhazardous, and unknown materials to either the ground surface, the Waste Water Treatment
 Facility (WWTF), or to containment structures.  According to LP&P personnel, ME DEP personnel
 were notified of each release. Not all releases had adverse effects (LP&P, 2003).

 In March of 1990, a Superior Court judge approved a settlement requiring Georgia-Pacific Corp. to
 pay a state record fine of $637,000 for alleged water and air pollution violations dating back to 1986.
 The civil fine against Georgia-Pacific, which became Maine's largest landowner when it recently
 assumed the property holdings of takeover target Great Northern Nekoosa Corp., is the largest ever
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Final RARE Report August 2015

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for violating Maine environmental laws, according to the Maine Department of Environmental
Protection (ME DEP).

                             Previous Investigations
The Penobscot Indian Nation established a Department of Natural Resources (PIN-DNR) in 1980 to
monitor and promulgate tribal ordinances to protect the Tribe's natural resources. Over the past three
decades, the Department has established model air and water monitoring programs collaborating with
local, state and federal partners to monitor the health of the ecosystem, develop toxicity studies
assessing the level of toxins Penobscot tribal members are exposed to, and restoration initiatives to
protect and restore the health of the ecosystem.

Monitoring of chemical contamination of the Penobscot River includes the State of Maine's Dioxin
Monitoring Program, established in 1988, and the Surface Water Ambient Toxics Monitoring
(SWAT) Program, established in 1993. The SWAT program includes monitoring for Dioxin,
Mercury and PCBs.  See http://www.maine.gov/dep/water/monitoring/toxics/.  In addition, the Maine
Department of Environmental Protection (ME DEP) maintains four sampling stations in the study
area as part of a statewide Dioxin Monitoring Program.  The State's Dioxin Monitoring Program
involves regular sampling and reporting of concentrations of dioxins/furans in fish, wastewaters,
sludges and effluents in the State of Maine (USEPA, 1989; Frakes, 1990; Opperhuizen, 1990;
Mower, 1991-2002; ENSR Consulting and Engineering, 1995; USF&W, 1996; ME DEP, 1999-
2000).

In 1998, the BIA approached EPA with their concern regarding the public health problems of the
Penobscot tribal members. BIA was concerned that the pollution discharges into the Penobscot River
were impacting the health of the Penobscot Nation. The BIA expressed concern with polychlorinated-
/>-dioxins (referred to as dioxin or PCDD in this document) and polychlorinated dibenzofurans
(referred to as furan or PCDF in this document), and polychlorinated biphenyls (PCBs) in fish and
sediment in the Penobscot River between the towns of Lincoln and Old Town, Maine.  As a result of
the BIA's concern, the BIA commenced an occurrence and distribution study of dioxin, PCBs and
furans in the Penobscot River in collaboration with the PIN, EPA, USGS, ATSDR and USF&W in
1998. The purpose of this study was to characterize the riverbed sediments in an effort to complement
the ME DEP data and more completely determine the ecological and human-health risks associated
with dioxin, furans, and PCBs to the PIN tribal members. A Quality-Assurance Project Plan (QAPP)
was developed by the cooperating agencies, authored by the USGS, and approved by EPA in 2001.
Samples of riverbed sediment from nine river reaches and fish from two river reaches between Old
Town and Grindstone, Maine were collected.  The University of Maine Environmental Chemistry
Laboratory (UMAECL) analyzed the samples for quantitative determination of dioxin, furan, and
PCB congeners.  However, due to a loss of funding and loss of staff, the lab equipment was not
properly maintained and the integrity of the samples collected were impaired.  USEPA conducted the
data validation and determined that the data were not of the quality specified in the QAPP. On April
16, 2003 EPA issued a letter to UMAECL that it was unable to accept the dioxin, furan, and PCB
analytical  results. EPA concluded that the data were not of sufficient quality to be used to report with
any degree of certainty concentrations of the compounds in the samples collected for the study.

In 1999, the U.S. Bureau of Indian Affairs submitted two requests to USEPA Region 1. The first letter
was received on February 18, 1999 requesting that EPA conduct a comprehensive multimedia
environmental compliance inspection of Lincoln Pulp & Paper Co., Inc. The second letter was received
on April 22, 1999 from Franklin Keel, the Director of the Eastern Area Office of the BIA. Director Keel


   Page   18                                           Final RARE Report August 2015

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requested that due to the numerous releases of hazardous substances in the Penobscot River, USEPA take
the necessary action to compile the appropriate information in to the CERCLIS system.

In response to BIA's request, in May of 1999, EPA conducted a multi-media inspection of the facility
and conducted a Preliminary Assessment/Site Investigation (PA/SI) of the Lincoln Pulp & Paper
Company. According to the PA/SI some of the previous investigations included a solid waste disposal
area investigation conducted by E.G. Jordan, Inc. in 1988 and Phase I and Phase II hydrogeological
investigations conducted by Sevee and Maher Engineers, Inc. in  1991 and 1995, respectively (Final
Preliminary Assessment/Site Inspection Report For Lincoln Pulp & Paper Co. Lincoln, Maine, May 30,
2003). Soil/source and sediment/source samples collected by EPA during this PA/SI indicated the
presence of three volatile organic compounds (VOCs), ten semivolatile organic compounds (SVOCs),
seven pesticides, three PCBs, 17 dioxins/furans congeners, 11 coplanar PCB congeners, and 14 priority
pollutant metals. Beryllium was detected in one sample in excess of the Maine State Remediation
Guidelines.

The PA/SI analytical results of the eight sediment samples collected from the Penobscot River
indicated elevated concentrations of one SVOC (fluoranthene), one PCB (Aroclor 1254), two metals
(copper and mercury),  nine dioxin/furan congeners, and 11 coplanar PCB congeners. Analytical
results of four sediment samples collected from Mattanawcook Stream indicated that one element
(mercury), eight dioxin/furan congeners, and ten dioxin-like PCB congeners were detected above
reference criteria in samples collected from areas downstream of the mill complex along
Mattanawcook Stream. Five dioxin/furan congeners and nine coplanar PCB congeners were detected
above reference criteria within the one unnamed stream sample (99-SD-03) (LP&P, 2003). Based on
analytical results of the EPA sediment samples, release of hazardous substances to Mattanawcook
Stream, the unnamed stream, and to the Penobscot River were documented.

On August 22, 2003, the EPA issued a letter stating that based on the available data and information
concerning the site condition, that the appropriate designation for the site was a "No Further Federal
Remedial Action Planned" (NFRAP) designation. EPA noted that its decision was based in part on
knowledge that the, "... the ME DEP has been working with LP&P over a long period of time to
address both solid waste & other issues at the property." EPA did state that its decision was subject to
revision in consultation with the Penobscot Indian Nation or the State of Maine based upon new
information or substantially altered site conditions. (Final Preliminary Assessment/Site Inspection
Report For Lincoln Pulp & Paper Co. Lincoln, Maine, May 30, 2003).
   Page  19                                                                     201

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                      Purpose and Objectives of Research
 Due to the variety of pollutants that are discharged into the Penobscot River (See Figure 3.), the
 Penobscot Indian Nation (PIN) questions the ecological health and water quality of the river and
 how this may affect the practices that sustain their way of life. As a riverine tribe, the Penobscot
 culture and traditions are inextricably tied to the Penobscot River watershed.  It is through hunting,
 fishing, trapping, gathering and making baskets, pottery, moccasins, birch-bark canoes and other
 traditional practices that the Penobscot culture and people are sustained.

 Unfortunately, EPA lacked exposure information for assessing health risks for New England Tribal
 Nations that are sustaining a tribal subsistence way of life.  The traditional methodology for health
 risk assessment used by the U. S. Environmental Protection Agency (EPA) is based on the use of
 exposure assumptions (e.g. exposure duration, food ingestion rate, body weight, etc.) that represent
 the entire American population, either as a central tendency exposure (e.g. average, median) or as a
 reasonable maximum exposure (e.g. 95% upper confidence limit). Therefore, EPA did not have
 means for assisting Federally Recognized Indian Tribes with developing Environmental  and Health
 Protection Policies in Indian Country to protect tribal members who live according to their unique
 Native American traditions.

 This study provides a scientific basis for the Penobscot Indian Nation for developing environmental
 and health protection policies that will protect tribal members who live according to their unique
 culture and tradition.  This RARE study characterizes the potential health risks from cultural
 practices of Penobscot Indian Nation tribal members. This preliminary risk assessment evaluates the
 potential for exposure and risk to Penobscot Indian Nation tribal members from contaminants in
 sediment and biota when gathering, hunting or fishing according to the PIN's treaty protected rights
 afforded to them by the U.S. Congress.

 Accordingly, the goal of this research was to assess potential exposures to dioxins, furans, PCBs,
 and mercury from ingestion offish, duck, turtle, medicinal plants, and ingestion and dermal contact
 of sediments in the absence of any remedial action within the study area. This study was a
 preliminary risk assessment designed to determine if contaminant concentrations in fish, snapping
 turtle, wood ducks, and plants in Regions of the Penobscot River relevant to where PIN tribal
 members hunt, fish and gather plants are a health concern. This study was not designed to be a
 statistically validated assessment of contaminant differences among study sites or among species.

Objectives:
   1.   Develop culturally sensitive methodologies for assessing the potential level of exposure
       Penobscot Indian Nation tribal members may have from maintaining tribal sustenance
       practices.
   2.   Conduct field surveys and laboratory analysis on targeted flora and fauna for chemical
       exposure to dioxins/furans, PCBs, mercury and methyl-mercury.
   3.   Assist the ATSDR by providing the necessary data to conduct a Public Health Assessment.
   4.   Establish protocols for assessing the level of exposure to PCBs, dioxins/furans and mercury to
       tribal members as a consequence of gathering tribal plants for medicinal and nutritional
       purposes; as well as, consuming fish, eel, wood duck, and snapping turtle as a primary source
       of nutrition.
   5.   Survey surface water, sediment, and drinking water from the Penobscot River and Indian
       Island to assess the potential exposure of Penobscot Indian Nation tribal members to
       environmental genotoxicants that continue cultural sustenance practices.
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 Since an ecological risk assessment could not be conducted, the ecological samples collected by this
 study were archived for potential future analysis.

                             Research Responsibilities
The USEPA, USGS, ATSDR, and USF&W collaborated with the Penobscot Indian Nation to design
a contaminant sampling and analysis program that would be usable for human health and ecological
risk assessment. Various funding opportunities were pursued and in 2007, the team was awarded
$100,000 through the Region 1 Regional Applied Research Effort (RARE) Competition.  The
purpose of RARE funding is to address priority research problems in EPA New England. The funds
allowed the EPA to partner with the Penobscot Indian Nation and other Agencies to conduct a
preliminary risk assessment of the Penobscot River Ecosystem.

An additional $30,000 in EPA RARE funds was acquired to conduct a supplemental study assessing the
exposure of Penobscot Indian Nation tribal members to environmental genotoxicants.  This
supplemental study used the bacterial Ames test for mutagenic testing of the surface water, sediment,
and drinking water from the Penobscot River near Indian Island, Maine.  Mutagenicity is a feature of
some cancer-causing (carcinogenic) chemicals.

Additional funds were also provided by Penobscot Indian Nation through EPA Indian Environmental
General Assistance Program (GAP), CWA Section 104(b) 3, and CWA Section 106 grants to support
sample collection and analyses.
 Left to right: Jan Paul, Jason Mitchell, Robert Lent, Dan Kusnierz, Gary Perlman, Valerie Marshal, Janet
Diliberto, Robert Hillger, Thomas Hughes, Robert Dudley, Carl Orazio, Jason Sockbeson
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                              Research Team
PIN DNR staff and Federal Partners listening to Dan Kusnierz at boat launch on the Penobscot River
This project involved a significant collaboration among several Federal partners and the Penobscot
Indian Nation. It involved the collaboration of approximately 50 scientists. See Appendix D:
Personnel Associated with RARE Study.

Penobscot Indian Nation (PIN)
   The Penobscot Indian Nation collaborated with USGS and USF&WS for logistical assistance with
   field collections of the flora, fauna, sediment, and water samples. USF&WS, USGS, and USEPA
   assisted the Penobscot Indian Nation with collecting the samples according to the approved QAPP.
   The Penobscot Natural Resource Department was the liaison to the tribal elders and facilitated
   consultations with the Penobscot Tribal Nation to assure that the tribe's unique traditional
   practices and lifestyle were accurately reflected and evaluated in this study.

United States Geological Survey (USGS)
   USGS Columbia Environmental Research Center (CERC)
   USGS was the lead agency for developing a Quality Assurance Project Plan (QAPP) for this study.
   The QAPP was developed in partnership with the Penobscot Indian Nation, USEPA, ATSDR,
   USF&WS, and the BIA. The USGS CERC lab  conducted the Congener-Specific PCB Analytical
   Process for the study and the TOC, grain size, and mercury testing for sediment.

   USGS Maine Water Science Center
   The Field Sampling Leader,  USGS, was the primary contact between the sampling team and the
   laboratories that conducted the processing and/or analysis. The USGS was the lead for the field
   sampling team (USGS, PIN-DNR, USF&WS, and EPA Region  1 and ORD) and was responsible
   for scheduling project fieldwork; establishing sampling site locations; organizing and coordinating
   shipping and handling with EPA NERL, USGS-CERC, Frontier Geoscience Lab, and EPA OPP
   laboratory managers; organizing and coordinating overall sampling  schedule with EPA NERL and
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Final RARE Report August 2015

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   PIN-DNR managers. Analytical chemistry data from each laboratory were reviewed by each
   laboratory's QA/QC program.

 United States Environmental Protection Agency (EPA)
   US EPA New England Region 1
   EPA New England Indian Program supported the submission of this RARE proposal and approved
   Valerie Marshall to participate as a co-lead with Janet Diliberto of EPA ORD for this project. As a
   leader of this project, Valerie Marshall led all the conference calls and meetings for this study,
   assisted with the fish sample collection, participated in all the QA audits, and facilitated the
   successful development and completion of this research project. EPA New England Superfund
   Program assisted with the scoping of the current  study and the review of the data generated.  EPA
   GIS Department assisted with developing maps for this study.

   US EPA New England Regional Laboratory at North Chelmsford, MA
   EPA New England Regional Laboratory (NERL) at North Chelmsford, MA  assisted with fish and
   turtle sample collection, development of the study and the QAPP, and conducted mercury sample
   analysis for the fish tissue. EPA New England's  Quality Assurance Program approved the Quality
   Assurance Plan for this project. Analytical chemistry data from each laboratory were reviewed by
   each laboratory's QA/QC program and then reviewed by the RARE Project Data Validator at
   EPA'sNERL.

   US EPA Environmental Chemistry Laboratory at the Stennis Space Center, MS
   The EPA Environmental Chemistry Laboratory (ECL), under the Office of Pesticide Programs
   (OPP) commonly referred to as the Stennis Lab conducted the analysis of PCBs, dioxin/furans and
   mercury of the sediment samples, and played an integral role in developing the parameters of this
   study and the QAPP and they also assisted with the review and validation of the sample analyses.

   US EPA/ORD at Research Triangle Park, NC
   Janet J. Diliberto was the co-lead for this project and assisted with the development of the QAPP,
   management of the Project, attended all site visits and participated in all the  QA audits.  Janet
   Diliberto retired from EPA on October 1, 2011.

   US EPA/ORD/NHEERL/RCU
   The Quality Assurance Manager was responsible for reviewing and approving the QAPP and
   served as the lead QA Auditor on the QA Audits. The Technical Systems Audit (TSA) of the
   Ames testing was conducted by EPA NHEERL QA Manager.

    US EPA ORD/EERD: Ecological Exposure Research Department was responsible for
      processing all the fish tissue samples collected.

Agency for Toxic Substances and Disease Registry  (ATSDR)
    This study involved collaborating with the ATSDR to assure the scope and  procedures identified
    in the QAPP met the objectives of the Penobscot Indian Nation to quantify the risk associated
    with Penobscot tribal members carrying out their traditional practices and ensuring that the
    methodologies employed are accepted within the scientific community. ATSDR participated in
    the scoping of this study and served as an integral research partner in performing and carrying out
    this research study. ATSDR attended all conference calls and meetings and assisted with the
    review and evaluation of the data generated from this study.  ATSDR conducted a Public Health
    Assessment on behalf of the Penobscot Indian Nation based on past data acquired and the data
    generated from this study.

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US Fish and Wildlife Service (USF& WS)
   The USF&WS has been conducting investigations on the Penobscot River examining
   contaminant residues in Atlantic salmon, sturgeon, and bald eagles, and endocrine disruption in
   smallmouth bass.  USF&WS' environmental contaminants biologist in the Maine Field Office -
   Ecological  Services, assisted in field collections for the RARE project.

 Bureau of Indian Affairs (BIA)
   The Penobscot Indian Nation also collaborated with the Bureau of Indian Affairs on this project
   to assure that the data generated is usable to link science to policy and decision-making for the
   Penobscot Indian Nation.  The Bureau of Indian Affairs participated in the scoping of this study.
                                       Approach
 Penobscot River
The study was designed as a preliminary risk assessment to determine if contaminant concentrations
in sediment, fish, turtle, wood ducks and plants in regions of the Penobscot River relevant to where
the PIN hunt,  fish and gather plants were high enough to be a health concern. Several meetings at the
Penobscot Nation Natural Resource Department office and numerous conference calls were held to
develop a preliminary screening for assessing the level of exposure concentrations to PCBs,
dioxins/furans, mercury, and methyl-mercury in sediment, plants, fish, duck and turtle from areas
commonly used by Penobscot tribal members when gathering, hunting and fishing in the Penobscot
River. Information was also gathered as to how the PIN tribal members consume the species collected
and what portion is typically consumed or used. For example, most PIN tribal members prepare the
sampled species by fileting and skinning them. Therefore, that is the sample portion we analyzed in
this study. Details of the specific collection and preparation method for each species is contained in
the Sample  collection design section of this report.

The team selected specific geographical locations (reaches) along the Penobscot River for flora,
fauna, and sediment collection. Six reaches deemed ecologically representative were selected along
87 miles of the Penobscot River between Old Town and Medway, Maine. The reaches were chosen
based on the sediment mapping conducted by USGS in 1999  (selected reaches of the study area
were mapped  during a bed-sediment mapping effort in May of 1999 [Dudley and Giffen, 2001]).  To
ensure the highest quality and reliability, a process of internal and external peer review by both
cultural and scientific experts was followed. The approach for this research project combined some of
the elements of consumption surveys such as interviewing Tribal elders to determine recent natural
   Page  24
Final RARE Report August 2015

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resource utilization patterns with careful identification of Tribal exposure factors (contaminant
concentrations, pathways of exposure). Collaborating with numerous scientists assured the scope and
procedures identified for this project met the objectives of the PIN and that the methodologies
employed are accepted within the scientific community.

With the flora and fauna data collected from this study, ATSDR conducted a Public Health
Assessment for the Penobscot Indian Nation that ATSDR will publish separately from this report.
EPA conducted a preliminary risk assessment by comparing the concentrations in biota to risk-based
concentrations to determine the level of risk to the Penobscot Indian Nation tribal members that
maintain cultural practices and sustenance lifeways associated with the Penobscot River. ATSDR
and EPA used Maine tribal ingestion and dermal contact rates that were developed in 2009.  Through
collaboration between EPA Region 1 and the federally recognized Maine Tribal Nations, exposure
scenarios that reflect the Maine tribal traditional cultural uses of natural resources were developed,
i.e. The Wabanaki Traditional Cultural Lifeways Exposure Scenario (Harper and Ranco, 2009).

The team also selected 4 sites for collecting surface water, sediment, and drinking water for the
Salmonella mutagenicity assays. The Salmonella mutagenicity assay was used to assess complex,
organic extracts of the river water, sediment from the river, and drinking water associated with the
PIN. The data collected from the Salmonella mutagenicity assays provide an integrated measure of
the mutagenic activity and, thus, potential carcinogenic activity, of the organics in the river water,
sediment from the river, and drinking water associated with the PIN.


                            Research Study Location
River reaches in this study (Figures 4-8) are demarcated upon the basis of river-features that control
or potentially impact fish passage or habitat (dam structures, impoundments, fish ladders, falls).
USGS field sampling leader, Rob Dudley, and the Penobscot Indian Nation Water Resource
Manager, Dan Kusnierz, divided the study  area into six sampling reaches associated with the
Penobscot Indian Nation Reservation distinguished by general hydrology and whether the area is
located in impounded waters of a dam  or free-flowing.  Three of the reaches (reaches 1-3) were
surveyed previously in 1995-96 and 1999 (Dudley and Giffen, 2001).

The reaches were chosen based on a sediment mapping study conducted by USGS in May 1999 in
which bed sediments were mapped in selected reaches of the study area (Dudley and Giffen, 2001).
The 1999 mapping effort involved the  use of ground-penetrating radar data to characterize the bed-
sediment composition in selected reaches of the Penobscot River. Sampling locations were chosen on
the basis of the mapping information and other river characteristics including wading and swimming
areas; deposit!onal zones within the channel; and  sites upstream and downstream of river features that
control or potentially impact sediment transport (such as dam structures and impoundments). The
control reach included both free flowing (East Branch Penobscot) and natural lake waters (Salmon
Stream Lake) that were within the upper Penobscot Watershed and upstream of any discharge or
known pollution sources.

Three types of samples were collected for the Salmonella mutagenicity assay: drinking (tap) water
from Indian Island, surface water from the  Penobscot River, and sediments from the Penobscot River.
River water and sediments were collected at the following locations: (1) an upstream  site (Salmon
Stream Lake); (2) a site slightly downstream of an industrial-outfall (Lincoln Paper and Tissue Mill);
and, (3) a publically owned treatment-works facility (Lincoln POTW), and a downstream site (West
Enfield impoundment).  A fourth sediment sample was obtained from a site adjacent to Indian Island.
Drinking water samples were collected at the PIN DNR, Water Quality Monitoring Laboratory on

   Page  25                                            Final RARE Report August 2015

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Indian Island, ME. Figure 8 and Table 2 provide details about the sites and the samples collected for
the mutagenicity analysis.
   Page  26
201

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                                        Reach 6: Control Reach, East
                                        Branch-Salmon Stream (EBS)
                                            Reach 5: Mattaseunk Dam
                                            Impoundment (MAT)
                               Reach 4:
                         Mohawk-Mattaseunk
                             Dam (MM)
                                Reach 3: West Enfield
                                Dam Impoundment (WEI)

                               Reach 2: Sunkhaze-West Enfield Dam (SWE)
                              Reach 1: Milford Dam Impoundment (MIL)
                                    •'

Figure 4: Penobscot River Study Six Reaches. The six reaches of the Penobscot River studied in this project.
   Page  27
Final RARE Report August 2015

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Figure 5: Penobscot River Study Reaches 1-2.
   Page  28
Final RARE Report August 2015

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012

Reach 4
734 Miles

Figure 6: Penobscot River Study Reaches 3-4.
   Page  29
Final RARE Report August 2015

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                                                                •
                                                    I
                                                            - •   *-TL/  <
                                                    S.        .     X*\ ;
                                                    J.^N-v-.. '. .   . -»  • I f
Figure 7: Penobscot River Study Reaches 5-6
     Page  30
Final RARE Report August 2015

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                                                                Pcnobscot River

                                                     Mutagenicity Sampling Locations
             Plymouth
           Water Quality Sample Type:


           O Drinking water   nRiverwater   A Sediment

         Inset maps drawn at 1:24,000 scale, except Salmon Stream Lake at 1:40,000
         Data Sources: Water Quality Sampling Locations: Penobscot Nation, EPA
         Base map © 2009 Microsoft Corporation and its data suppliers. EPA R1 GIS map# 8525_2
Figure 8:  Mutagencity drinking water, surface water, and sediment sample collection sites.
     Page   31
Final RARE Report August 2015

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                  Sample Collection Design and Analysis
  From Left: Robert Lent, USGS; Robert Hillger, USEPA; Jason Sockbeson, PIN DNR; Gary Perlman,
  ATSDR; Valerie Marshall, USEPA.

Research Project Schedule
The funding for this research study was awarded in June 2007. Following the development of the
QAPP and the completion of the design criteria for the study, samples were collected and analyzed
from May 2008 until January 2011.  Throughout the research period several  audits were conducted to
ensure the integrity of data collection and management. The consolidation and final analysis of data
results were completed by September 2011. An overview of the project schedule can be found in
Appendix C.

Sample Collection and Preparation
The study area was divided into six reaches, including a control reach. The reaches were identified
by USGS in collaboration with the Penobscot Indian Nation, and are generally demarcated on the
basis of river features that affect fish passage or habitat (dams, impoundments, free-flowing) (Figures
4 - 7). The control reach is upstream of any point source discharges and includes both free-flowing
riverine habitat (East Branch Penobscot River) and natural lake habitat (Salmon Stream Lake). A
variety of flora and fauna species, and sediment samples from each of the six reaches were collected.
Some plants that were collected are used for medicinal purposes by the Penobscot Indian Nation and
cannot be named in this report. To protect these resources from being exploited, these plants are
referred to as "medicinal plants". The following section contains a description of the species collected
and how they were sampled. An overview of the quantity, type, and species collected within each
reach is provided in Table 1.  All field sample  collection and sample handling followed explicit
protocols outlined in the QAPP.
   Page  32
Final RARE Report August 2015

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Because the PIN has a concern about the possibility of environmentally induced cancers among their
population caused by municipal and industrial waste discharges into the Penobscot River, the project
team determined that an important aspect of this study should include evaluating if environmental
mutagens occur in surface water, sediment and drinking water because mutagenicity could be
indicative of the presence of potential carcinogens. To address these concerns, monitoring methods
using the Salmonella mutagenicity assay (Ames test) were employed to test for mutagenic
activity. Because there are many classes of carcinogens e.g., metals, fibers, dyes, and certain
polycyclic aromatic hydrocarbons (PAHs), efforts to establish the presence or absence of
mutagenicity in these samples serves to evaluate whether additional studies on potential carcinogens
may be needed.

Samples Collected
Reaches were sampled from May 2008 to October 2009. Quality-control (QC) duplicate samples
were obtained. Each sample location was recorded using a hand-held GPS unit. All sampling
procedures followed USEPA-approved protocols as outlined in the QAPP for this project (Orazio,
2008). Daily field logs were maintained. The  surface water,  drinking water and sediment samples for
the Mutagenicity assays were collected from July-October 2009. QAQC procedures were followed
for collecting these Samples (See Claxton and DeMarinin EPA Intramural Research Protocol). Tables
1 and 2 below summarize the samples collected for this study.
   Page  33                                            Final RARE Report August 2015

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Summary of Composite Samples Collected of Sediment, Flora, and Fauna
Reach

1
(MIL)
2
(SWE)
3
(WEI)
4
(MM)
5
(MAT)
6
(EBS)
Sediment
n
(water depth)
3 shallow
(0.6-0.9m)
1
impoundment
(1.8m)
3 shallow
(0.6 - 0.8m)
3 shallow
(0.8-1. 1m)
1
impoundment
(2.4m)
3 shallow
(0.3- 0.6m)
3 shallow
(0.9m)
1
impoundment
(6.7m)
3 shallow
(0.5 - 0.6m)
Perch
n (spp.)
#/Wt. (kg)
Length (mm)
1 (White)
7/2.26
245-300
1 (Yellow)
13/1.76
204-270
1 (Yellow)
14/2.42
221-280
1 (White)
6/1.16
188-270
1 (Yellow)
5/.37
151-208
2 (White)
8/2.23
249-289
8/2.19
249-271
1 (White)
8/2.93
273-297
1 (Yellow)
14/2.51
208-265
8MB
n
#/Wt.(kg)
Length (mm)
1
4/2.40
320-405
1
5/3.5
347-464
1
4/3.22
328-443
2
4/3.91
406-440
4/3.74
387-423
1
4/2.05
338-362
1
4/4.77
390-449
Bullhead
n
#/Wt.(kg)
Length (mm)
1
11/2.39
220-284
1
10/2.89
248-328
1
12/2.12
219-288
1
9/2.02
205-288
1
3/0.73
206-290
1
8/3.3
270-340
Pickerel
n
#/Wt.(kg)
Length (mm)
1
8/2.15
335-410
1
7/2.27
350-431
1
6/2.16
342-447
1
7/2.34
332-433
1
6/2.73
397-487
1
7/3.21
382-500
Eel
n
#/Wt.(hg)
Length (mm)
1
4/2.07
555-721
1
5/1.75
481-684
1
4/2.7
632-790
1
4/2.47
606-761
1
4/1.86
576-684
1
2/0.52
509-556
OSF*
n
#/Wt.(hg)
1
3/2.67
2
3/1.87
3/1.87
1
3/2.15
2
3/1.59
3/1.48
0
1
3/3.02
Med. Plant **
n
#/Wt. (kg)
0
0
1
3/0.48
1
3/0.5
1
3/0.51
2
3/0.63
3/0.62
Duck
n
#/Wt.(k
g)
1
4/0.79
1
3/0.58
2
3/0.57
3/0.59
1
3/0.81
0
0
Turtle
n
#/Wt.(k
g)
1
1/2.76
0
1
1/2.64
2
1/2.95
1/1.98
1
1/2.61
2
1/1.0
1/2.35
Table 1: Summary of composite samples collected and analyzed for the project. For non-sediment samples, n = number of composite samples, # = number
of individuals or sites used in composite, wt. = total wet weight of tissue in composite. *Ostrich Fern (OSF). ** Medicinal Plants (Med. Plant).
        Page  34
Final RARE Report August 2015

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r Summary of Samples collected
for
Mutagenicity Assays

Drinking water
River Water
River Sediment
# of samples
3 (composites)
9 (composites)
4 (composites of
top sediment (<15
cm depth))
Assay
Salmonella mutagenicity
assay
Salmonella mutagenicity
assay
Salmonella mutagenicity
assay
Table 2:  Summary of samples collected for Mutagenicity Assays.
 Sediment Collection and Preparation
 Robert W. Dudley, USGS, collecting sediment sampling
  Twenty one sediment samples were collected in July 2008. Three shallow water sediment composite
  samples, comprised of 2-5 grabs, were collected within each reach. Wading areas were chosen based
  upon the following criteria: Deposit!onal zones of fine-grained material (both sand/silt and materials
  potentially rich in organic content) observed within the river channel via geophysical techniques; and,
  known or suspected littoral wading-contact areas along the mainland and island shorelines (associated
  with swimming, hunting, fishing, plant harvesting, boat launching, etc.).

  Because the Penobscot River is a relatively high energy river system with frequent flushing of
  sediments, the team believed it was important to also analyze sediments behind some of the
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Final RARE Report August 2015

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impoundments (dams) where sediments have accumulated over time. One deep water composite sample
was collected from each of the impoundments behind the Milford, West Enfield and Mattaseunk dams.

At each sampling location multiple grabs (2-5) of surface sediments (0-6" deep) were collected using a
Ponar dredge sampler. After allowing excess water to drain, the sediment grabs were placed in a large
metal pan until sufficient sample volume was collected.   The sediment grabs were thoroughly mixed in
a metal bowl using a spoon until homogenous and then transferred into labeled amber glass sample
containers.  Individual sample containers were placed in sealed Ziploc® bags and placed in coolers
with double bagged water ice for transport to the PIN laboratory.  Sediment samples were stored in a
refrigerator near 4°C until they were shipped with ice to each laboratory for analyses.
 Fish Collection and Preparation
Collection offish at the Lincoln, Maine boat dock on the Penobscot River
                         US EPA Robert Hillger displaying eel trap used for this study at PIN DNR
Thirty-four composite fish samples, representing 228 individual fish from six species were collected from
July - October 2008. The goal was to collect five species from each reach: Smallmouth bass (Micropterus
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dolomieu), chain pickerel (Esox niger\ white perch (Morone americana) or yellow perch (Perca
flavescens) [depending on which perch species was present in the reach], brown bullhead (Ameiurus
nebulosus) and American eel (Anguilla rostratd). Fish were collected by angling with line and tackle, trap
nets, gill nets, or boat electro-shocking.  Single species composite samples for each fish species of a reach
was created by combining 3-5 individuals or as many as were needed to obtain a total mass of > 2 kg.
White and yellow perch tissues were not composited together. The target size of fish was what is typically
kept and consumed by Tribal members (See Table  1).  The goal was to use fish of similar size so that the
smallest individual in a species composite was no less than 70% of the largest individual (length). In the
few instances in which the team was unable to meet this goal, the sample weight was recorded and the fish
tissues were processed anyway.  In these cases an attempt was made to analyze the samples for as many
contaminants as possible with the limited sample amount.

 Field duplicate pairs for fish consisted of two composite samples taken at a site containing fish of similar
length. At two river reaches we produced paired composite samples offish (of the same species) in which
the fish sizes were as closely matched as possible. We chose white perch at Reach 5 (MAT) and bass at
Reach 4 (MM) because those were the species and  reaches for which we had an abundant supply of fish
and from which we could make composites of similar size fish.

As fish of the appropriate species and approximate size range were collected, they were kept alive in
coolers containing water until they were killed.  Initial processing offish was done in designated areas
according to the approved QAPP.  The field sampling team measured and recorded length and weight.
Whole body fish were double wrapped in aluminum foil, labeled, placed in a Ziploc® bag and frozen in
a secure location at the PIN laboratory.  Fish were stored frozen until a sufficient number and size of
fish were accumulated. Individual fish to be composited were organized into a plastic bag and labeled
according to the Sample Labeling Protocol. Fish were shipped frozen in coolers containing dry ice to
Dynamac, ORD's on site contractor, for filleting.

Because we were informed that most tribal members prepared the fish species used in the study by
skinning and filleting, we used skinless fillets. Dynamac filleted and skinned each fish according to the
approved research procedures with the following modification; fish were slightly thawed before
removing skinned and boneless fillet with a sharp fillet knife or scalpel. The weight of the two removed
fillets and the weight of the remaining offal (including skin and bones removed during filleting) from
each fish were recorded separately. The fillets and offal from those samples collected at each reach
were  composited separately into two distinct samples. Skinned and boneless fillets were prepared for
smallmouth bass, pickerel, perch, and bullhead.  Sections of eel were taken after they were cleaned and
skinned. At least 500g of skinless fillet tissue was needed to conduct the various chemical analyses.

The fillets of individuals of a species from  a reach were wrapped in aluminum foil making sure that the
dull side was in contact with the fillet and shiny side was on the outside, placed in a pre-labeled
Ziploc® bag  (following Sample Labeling Protocol), and placed in a -20°C freezer until they were
shipped to the Region 1 laboratory. Once a cooler full of fillets had been accumulated, they were
shipped on dry ice to the Region 1 laboratory contact, Dave McDonald, for homogenization. The
remaining portion of the fish (referred to as offal or carcass) was wrapped in aluminum foil and shipped
on dry ice to  Joseph Ferrario/Stanley Mecomber at EPA Environmental Chemistry Laboratory and was
stored in labeled bags and frozen for potential future analysis for ecological risk assessment.
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Plant Collection and Preparation
i
Charles Culbertson, USGS
Washing ferns
Ostrich fern
                                                                   -
Robert Dudley, USGS
collecting ferns
Fiddlehead fern (ostrich fern, Matteuccia struthiopteris) and a medicinal plant were analyzed due to the
significance of these plants to the Penobscot Indian Nation culture and diet. Fiddleheads were collected
as they emerged from the soil in May 2008, when and at locations harvested by tribal members. The
medicinal plant was collected during early a by autumn (September and October 2009) when it is easiest
to identify and harvest by tribal members. While the plant is also sometimes harvested in the spring, it is
less abundant, more difficult to identify,  and less accessible at that time. While carrying out other
collection activities and with the assistance of a tribal botanist, the team located sites where the medicinal
plant could be harvested. The sites were revisited during September and October for collecting, while
taking care not to overharvest from any one area.  Per the USEPA approved QAPP, the data is stored as a
record from this study in Penobscot Indian Nation DNR files.

A total of seven composite fiddlehead samples were obtained, representing five of six reaches and one
field duplicate sample. We were unable  to find fiddleheads in the MAT Reach (Reach 2).  In the MM
reach (Reach 4) we collected two composite samples from the following sites; one downstream of the
Lincoln Paper and Tissue mill and another upstream in an area heavily utilized by tribal harvesters.

Fiddlehead ferns were collected using similar methods to those used by tribal members when harvesting.
We also analyzed that portion of the fern that is consumed by tribal  members. Fiddlehead ferns were
collected by breaking emerging fiddleheads from the stems and placing in a 1-gallon Ziploc® plastic bag
labeled with reach and location.  Approximately 500g of sample was collected at each site station
location.  Each reach composite sample was comprised of fiddleheads collected from two to three  site
station locations within the reach segment. The samples were transported in a cooler with double bagged
water ice to the office/PIN lab.  Soil and  non-edible brown skins were removed from the fiddleheads by
soaking and spraying the ferns with tap water through a screen. The ferns were then rinsed in deionized
(DI) water. The cleaned fiddleheads from each location within a reach were combined and thoroughly
mixed together. Approximately 500 grams of cleaned fiddleheads were weighed out with a balance or
scale and placed in a new Ziploc® bag with a sample labeled according to the Field Sample Numbering
protocol. Fiddlehead samples were frozen at -20°C.

A total of five composite medicinal plant samples were collected representing four reaches and one
duplicate. The type of plant tissue (leaves, stems, roots) that was composited represents what is typically
used by tribal members.
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The medicinal plant collected for this study was collected using the same methods used by tribal
members when gathering the plant. Root materials from the medicinal plants were extracted from where
it grows by digging with gloved hands, or with the use of a trowel.  The leaves were removed by cutting
with a sharp knife and the root materials were placed in a Ziploc® bag labeled with reach and location.
Depending upon abundance, each reach sample was comprised of root material from two locations within
each reach segment. Samples were transported in a cooler with double bagged water ice to the office/PIN
lab.  At the PIN lab, the root materials were rinsed with tap water to remove soil, and then rinsed with DI
water. Approximately equal amounts of plant material from each location within a reach were combined
and thoroughly mixed together. For each reach approximately 500g of root materials were weighed out
with a balance or scale and placed in a new Ziploc® bag with a sample label according to Field Sample
Numbering protocol.  The medicinal plant  samples were frozen at -20°C.

Field duplicate pairs were collected for fiddlehead (1 duplicate) and medicinal  plant (1 duplicate) samples
from one of the six reaches. Both samples of a field duplicate pair were a composite of plant materials
collected from the same locations within a reach and in accordance with the QAPP.

Plant samples were then shipped frozen from Maine on dry ice to CERC-USGS for analysis preparation.
Upon delivery to laboratories, chain-of-custody forms were signed and the samples were stored in a
secure location frozen at -20°C until processing.
Turtle Collection and Preparation
                                                                                        • -
USF&W photo
USF&WS Digital Library http://digitalmedia.fws.gov/cdm/
Snapping turtles (Chelydra serpentine) are an important tribal food. Initially the team had discussed
collecting three snapping turtles per reach; however, due to the concern of negatively impacting the
population, we reduced the targeted number of snapping turtle per reach.  A total of seven snapping
turtles were collected from five of the six reaches.  No samples were obtained from Reach 2.
In September 2008, the team collected snapping turtles from two of the six reaches.  In an attempt to
collect snapping turtle from all the reaches, the team decided to try to collect snapping turtle again in
2009. During July - September 2009 we successfully collected snapping turtles from five of the six
reaches. Snapping turtles were captured using baited hoop net traps. Snapping turtle traps were set up in
slow moving water in suitable turtle habitat.  Traps were staked, baited with fish from the local reach, and
set out overnight.  Snapping turtles larger than 5 Ibs. were collected because this is the size used by tribal
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members and the size the team estimated was needed to provide a sufficient amount of tissue (-500 g) for
analyses. Upon collection, each turtle was tagged with its unique identification number and placed in a
cooler of water ice and transported alive to the PIN office/lab. Each sampling location was identified in a
field book and the sample coordinates using the GPS system were also recorded.

Each specimen was weighed to the nearest 0.5 Ib with a hanging scale and measured (carapace length - to
nearest 2 mm).  Specimens were killed by decapitation after cooling them down in a freezer for several
hours to slow reflexes and induce torpor.  Specimens were held and processed within 48 hours of
collection. Animals were rinsed with tap and DI water to dislodge sediment or other external material
from their skin prior to making incisions.  All equipment was pre-cleaned and managed following the
QAPP guidance. We used muscle tissue portions that are typically consumed by tribal members. Using a
pre-cleaned knife and scalpel, muscle tissue was removed from the hind limbs, fore limbs, tail, and neck.
The muscle tissue was skinned and the bones removed. Adipose tissue deposits were not included with
the muscle tissue sample. The mass of the meat was weighed and recorded to the nearest 1 g. The tissue
was wrapped in an aluminum foil packet, placed in a pre-labeled Ziploc® bag (labeled following Sample
Labeling Protocol), and frozen to -20°C. Observations of internal or external anomalies and sex of the
specimen were recorded for each turtle. Turtle meat samples were shipped frozen from Maine on dry ice
to CERC-USGS for grinding and sample preparation.
Duck Collection and Preparation
              USF&WS Digital Library http://digitalmedia.fws.gov/cdm/
Although several species of duck have been historically important as food for tribal members, wood
ducks (Aix sponsa) are currently the most utilized local duck species on the Penobscot River. Because
wood ducks commonly breed on the Penobscot River, they are more likely to reflect local contaminant
levels than other species that use the river more seasonally.

Wood ducks were collected mid-September to early October 2008, the time when tribal members
typically hunt wood duck.  Sixteen individual wood ducks were collected, representing five composite
samples from four of the six river reaches. A duplicate sample was also collected from one reach. Ducks
were collected by shooting with a shotgun with steel shot ammunition.  The location of each collection
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 site was recorded on a map and GPS coordinates were recorded in a field notebook.  Reach composite
 samples were created by combining muscle tissue from three to four wood ducks from each reach to
 provide a total weight > 0.5kg. The collected birds were placed in labeled Ziploc® bags and transported
 on water ice back to the PIN laboratory for processing.

 At the lab, weight, age, and sex of each wood duck were measured and recorded. Muscle tissue from the
 breast and legs (boneless, featherless and skin-on) was excised from each duck using a knife or scalpel.
 The muscle samples reflect the portions that are typically consumed by Tribal members.  The excised
 tissue was weighed, wrapped in foil and Ziploc® bag, labeled and frozen.  Tissue samples were shipped
 frozen with dry ice to CERC for compositing, grinding, and sample preparation.  The remaining
 carcasses (including organs) were stored in labeled bags and frozen for potential future analysis for
 ecological risk assessment.

 Collection and Preparation of Surface  Water, Drinking Water, and Sediment
for Mutagenicity Analysis
 Water samples from the Penobscot River were obtained by collecting a composite of five sequentially
 filled 2.5-L bottles  at each location for each sampling event. Samples were collected in amber bottles
 that were pre-cleaned and had a Teflon™ cap (Cat. No. #293680, Sci Spec, Hanover, MD) by
 submerging the capped bottle within 0.3 m of the river's surface, uncapping the bottle until it was filled,
 and recapping the bottle under the water. Care was taken to avoid disturbing bottom sediments to keep
 them  from entering the sample bottle.  Samples were placed in a cooler with water ice in the field and
 then stored in a refrigerator at 4°C in the dark until they were shipped. In  order to keep samples cooled,
 they were shipped in coolers containing frozen Blue Ice®.

 Drinking water samples were collected by compositing five sequentially filled 2.5-L bottles for each
 sampling event from a convenient tap on Indian Island at the PIN Water Quality Monitoring Laboratory.
 One composite was taken on the same days that river water was sampled.  The water from a drinking
 water tap was allowed to flow for 10 min prior to collecting the samples. Samples were collected in
 amber bottles and stored in a cold room at 4°C.  The drinking water samples were shipped in the same
 manner as the river water samples.

 Sediment samples were collected at approximately the same sites as the river water was collected. Three
 river-sediment samples, were taken. At each location a composite of three to five grab samples of the top
 sediment (<  15cm) was taken using a Ponar dredge. The grab samples were placed in a stainless-steel
 container and mixed together until homogenous. The composite sample was then divided and transferred
 into 3 pre-cleaned amber glass jars with Teflon™ lined lids, each containing ~500g wet weight of
 sediment. The dredge and sampling equipment were  cleaned with Alconox, deionized water, and
 methanol before  and between sampling at each site. Each jar was placed in a plastic sealed bag and
 placed in a cooler with water ice for transport from the field until the bag was transferred to a dark 4°C
 refrigerator.  Sediment samples were shipped in coolers with Blue Ice®.
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                 Summary of Chemical Extraction Methods
Sediment/Vegetation
The sediment and vegetation samples collected from the Penobscot River watershed were extracted using
a similar procedure.  In addition, the plant material was prepared according to tribal cultural practice
(washed, dried, stemmed, etc.) prior to compositing. A weighed quantity of dried sediment or vegetation
was processed by means of Soxhlet extraction. The samples were weighed into a glass fiber thimble and
then mixed with a quantity of anhydrous sodium sulfate.  After the thimble was placed in the Soxhlet
extraction  column, the samples were spiked with a 13C PCDD/PCDF/co-planar PCB compound mixture
and extracted for a minimum of 12 hours. After extraction, the sample extracts were stirred with
acidified silica gel and then decanted. The sample extracts were then further cleaned by means of
acid/base chromatography.  Following the acid/base chromatography, the PCDDs/PCDFs were separated
from the co-planar PCBs using carbon column chromatography on ECL prepared columns using AX-21
(Winters et al., 1996; Ferrario et al., 1997).  The PCBs were concentrated to 20 jil or less for
FIRGC/HRMS analysis.  The PCDDs/PCDFs were further purified using alumina column
chromatography, followed by concentration to 20 jil or less and analyzed by HRGC/HRMS analysis.

Tissue
Tissue samples collected from the Penobscot River watershed were weighed into a Nalgene® bottle,
spiked with a 13C PCDD/PCDF/co-planar PCB compound mixture, and extracted three times. Each
extraction  consisted of Polytron® grinding, centrifugation, and filtering through anhydrous sodium
sulfate.  After the final extraction, the sample extracts were stirred with acidified silica gel and then
decanted. Additional fractionization, purification and analytical techniques were as described for
sediments/vegetation or can be found in subsequent sections. ECL has worked extensively on the EPA
Dioxin Reassessment Study and analyzed a number of food items including beef, pork, poultry, and milk
(USEPA, 2004).

Water and Sediments for Mutagenicity Analyses
All extracts for use in mutagenicity analyses were prepared in dimethyl sulfoxide (DMSO) as well as the
direct-acting controls 2-nitrofluorene and sodium azide (Sigma, St. Louis, MO, 3.0 ug/plate) and the
indirect-acting control 2-anthramine (Sigma, 0.5 ug/plate). The river-water samples were extracted by
open-column chromatography using a 50:50 layer of XAD-2/XAD-8 resin with the XAD-2 on the
bottom;  organics were eluted with ethyl acetate. The extracts were dried over sodium sulfate,
concentrated, filtered across a 0.45-um polytetrafluoroethylene (PTFE)-syringe filter, and solvent-
exchanged into DMSO at 5,OOOX for the bioassay (unless this was too thick, at which point more DMSO
was added to make the concentrate at 1,OOOX).

The drinking-water samples were processed as above except that the water was first acidified to pH 2
prior to extraction. Blanks prepared in the same way with XAD were also evaluated for mutagenicity.
River-sediment samples were processed by taking  100 g dry-weight of each sample and extracting each
by Accelerated Solvent Extraction (ASE) with a 50:50 mix of dichloromethane/methanol using an ASE
350 (Dionex Corp, Sunnyvale, CA). The extracts were filtered across 0.45-um Teflon™ laminated-filter
disks, concentrated, and solvent-exchanged into 1 ml of DMSO.
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                                   Sample Analysis
Contaminants of concern and the Data Quality Objectives (DQOs), Project Quantification Limits (PQLs)
and Targeted Laboratory Quantification (TLQ) limits for fish, turtles, ducks, plants, and sediment
collected from the Penobscot River can be found in the QAPP, dated May 13, 2008. The limits identified
in the QAPP were agreed upon by the Project Team.  The project team agreed to match the Project
Quantification Limits (PQLs) and Targeted Laboratory Quantification (TLQ) limits for fish, turtles,
ducks, plants, and sediment collected from the Penobscot River to the detection limits of the laboratory,
e.g. based on the capabilities of the laboratory equipment.  The detection limits were optimized by
maximizing the amount of sample analyzed, reducing background, concentrating the extracts to low final
volumes, and using highly sensitive instruments. For the high resolution gas chromatograph/high
resolution mass spectrometer used in the analysis of samples for PCDDs/PCDFs/co-planar PCBs, the
instrument operated at a resolution of 10,000 and a sensitivity of femtograms per gram (e.g. the resolution
has no units. The formula for resolution has the same units [atomic mass units (m/z)] in the numerator and the
denominator, so the units cancel.) The following table and text describe the types of analyses for the flora
and fauna of this study.
Summary of Sample Types Collected for Chemical Analyses.
Sample
Type
Sediment
Fish
Plants
Turtle
Meat
Duck
Meat
PCBs
X
X
X
X
X
Dioxins/
Furans
X
X
X
X
X
Total
Mercury
X
X
X
X
X
Methyl
Mercury
X

X
X
X
TOC
X

X


Grain
Size
X




Lipid

X

X
X
   Table 3: Summary of sample types collected for chemical analyses. This table includes the specific analyses
   performed on each sample type.  Samples for mutagenicity analysis are not included in this table.

Sediment
Twenty-one sediment samples were collected and analyzed for the twelve WHO (World Health
Organization) co-planar dioxin-like PCBs, dioxins/furans, total mercury, methyl-mercury, total organic
carbon (TOC), and grain size. The analysis of these sediments revealed low levels of co-planar PCBs. To
determine if the presence of the co-planar PCBs is directly related to the amount of total PCB congeners
present, a subset of nine sediment samples was analyzed for total PCB congeners. The three deep water
sediment samples collected were analyzed for grain-size distribution,  TOC, and concentrations of
dioxins, furans, co-planar PCBs, and total mercury.
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Fish Tissue
Fish tissue samples were processed for percent lipid, total mercury, dioxins, furans, and PCB congeners.
For PCB congener analysis in fish, six composite samples of smallmouth bass (8MB) were analyzed. The
study team chose 8MB composites for looking at patterns of PCBs in the fish as a
means of potentially identifying source(s). The smaller subset offish was tested for total PCBs congeners
based on the reasoning stated above for sediments. The team agreed not to analyze the fish samples for
methyl mercury.

In higher trophic level carnivorous fish, the ratio of methyl mercury to total mercury generally
approaches unity, meaning that almost all of the mercury in the fish fillet is in the methyl mercury form.
(See, Wiener et al., 2003)  It is generally assumed that >90% of mercury in higher trophic level
carnivorous fish is in the methyl mercury form, although site-specific variables and trophic level can
influence the ratio of methyl mercury to total mercury in fish. The team agreed that analyzing the
Penobscot fish fillet samples for total-mercury was a cost effective, accurate, and a slightly conservative
way to estimate the level of methyl mercury.

Turtle and Duck Tissue
The composited snapping turtle and wood duck composite samples were analyzed for total lipid, total
mercury, methyl mercury,  dioxins/furans, and PCB congeners. Samples were stored in a secure location
frozen at -20°C until processing began. Observations of internal or external anomalies were recorded for
each snapping turtle.

Plants
Plant composite samples were analyzed for dioxins and furans, PCB congeners,  methyl mercury and
mercury. Ferns and medicinal plants are approximately 90% and 75% water, respectively. Therefore,
each fern sample containing approximately 500 grams of fresh material produced -50 grams of dried
material; and, each medicinal plant sample produced ~ 125 grams of dried material. At CERC, samples
were homogenized with a Tissuemizer, freeze-dried, and aliquots were sent to the OPP, NERL, and FGS
for analysis.  Freeze-dried  samples were analyzed by 4 different labs as follows: 5 grams to NERL, 25
grams to EPA-OPP, 5 grams to FGS, and 10 grams retained at CERC.

                                 Analytical Procedures
Total Mercury
Fiddlehead fern, medicinal plants, snapping turtle, and fish tissue samples were analyzed for the presence
of total mercury (t Hg) by the Milestone DMA 80 laboratory at the EPA New England Regional
Laboratory.  All analyses were completed by the end of September 2010 and reported in October 2010.
Freeze  dried homogenized samples were analyzed using flash vaporization by a  Milestone DMA80
Mercury Analyzer. Samples were heated to 850 degrees centigrade to release all mercury from tissue.
Mercury vapor was then passed through a catalyst into an amalgamator for capture. After all the mercury
was captured, the amalgamator was heated to 200+ degrees centigrade and the released mercury vapor
was passed into a cuvette through which UV light was passed. A UV photodetector measures the
difference in UV light (mercury vapor absorbs light at 254nm).  The absorbance of UV is an indirect
measure of mercury concentration in the tissue. All total mercury analyses were carried out using the
EPA standard operating procedure (SOP) - "Milestone SOP2 (04/13/10) Standard Operating Procedure,
Mercury Analysis by Milestone DMA-80". All results are reported as dry weight in ug/kg.
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Methyl Mercury
Methyl mercury analyses were conducted by Frontier GeoSciences, Inc. (FGS) using FGS methods.
Homogenized tissue samples were digested for approximately 2-4 hours at 70-80 °C with a potassium
hydroxide and methanol solution.  After cooling, samples were diluted with methanol according to the
protocol - "Digestion of Tissue Samples for Methyl Mercury Determination - FGS-010". Sediment
samples were extracted using the protocol "Extraction of Soil or Sediment Samples for Methyl Mercury
Determination - FGS-045": Homogenized sediment samples were vigorously shaken for one hour with
methylene chloride and acidic bromide and copper sulfate solutions. After centrifugation and removal of
the aqueous layer,  an aliquot of methylene chloride was added to water and purged with nitrogen for
approximately thirty minutes to remove the methylene chloride. The sample, now in the aqueous phase,
was brought to final volume with reagent water.

Methyl mercury samples were analyzed using Cold Vapor Gas Chromatography Atomic Fluorescence
Spectrometry using protocol "Methyl Mercury Determination by Cold Vapor Gas Chromatography
Atomic Fluorescence Spectrometry (CV-GC-AFS) -  SOP FGS-070" Acetate buffer and ethylating agent
were added to an aliquot of digested sample and the methyl  mercury was purged onto carbotraps.  The
mercury species on the carbotrap column were volatilized and separated with a gas Chromatography
column, reduced on a pyrolytic column and then analyzed by thermal desorption into an atomic
fluorescence detector using the dual amalgamation technique. A chart recorder was used to record the
detector signal. Peak heights were  measured by hand and entered manually into the Laboratory
Information Management System (LEVIS) for calibration and calculation of concentration.

Congener-Specific PCBs (USGS-CERC)
The following series of USGS-CERC SOPs were used for analysis of the  sediment and fish samples for
congener specific PCBs:  SOP186, SOP187,  SOP270, SOP271, SOP461,  SOP464, SOP642, and
SOP643.  Several types of QC samples accompanied the analysis:  field/procedural blanks, matrix blanks,
matrix spikes, laboratory reference material, procedural recovery standards, and triplicate analyses. The
biological tissue sample composite was dehydrated with sodium sulfate. Sediment homogenates were air
dried and then were dehydrated with Na2SO4. The sample was spiked with procedural recovery
compounds PCB 029 (2,4,5-trichlorobiphenyl), PCB 155 (2,2',4,4',6,6'-hexachlorobiphenyl), andPCB
204 (2,2',3,4,4',5,6,6'-octachlorobiphenyl). The dried tissue samples were column extracted and
sediments were Soxhlet extracted,  both with dichloromethane (DCM).  Aliquots of extract were used for
percent lipid measurement. Bulk lipids and co-extracted biogenic materials were removed from the
extracts by low-pressure size exclusion Chromatography 70-cm SX-3 BioBead column with DCM mobile
phase, then by high-pressure size exclusion Chromatography Phenomenex 300 X 21.2-mm Phenogel 10
100 A column with DCM mobile phase. Elemental sulfur was removed from sediment using a
combination of copper and HPSEC treatments. Extracts were  cleaned up  with reactive adsorbent silica
gel columns, then fractionated using two layered octadecyl silica (ODS)/activated silica gel 60 (SG-60).
PCB congeners were measured by dual-column GC-ECD according to CERC  SOP P. 195. Analyses were
performed using cool on-column capillary injection onto retention gaps connected to 60 M DB-5 (5%
phenyl-, 95% methylsilicone) and DB-17 (50% phenyl-, 50% methylsilicone) analytical columns, or
equivalent.  Potential peaks for PCB congeners were matched and identified on one or both GC capillary
columns.  The capillary GC-ECD data were collected, archived in digital form, and processed using
Chromatography data system software.  Up to nine levels of calibration  for each individual congener were
used to quantify approximately 142 congeners. The calibration curve ranged from 10 to 8,000 ng/mL
total PCB concentration.
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Polychlorinated Dibenzo-p-dioxins (PCDDs), furans (PCDFs), and co-planar PCBs
[dioxins/furans/cp-PCBs](EPA/OPP/ECL)
The OPP Environmental Chemistry Laboratory tested the fish tissue samples for seven polychlorinated
dibenzo-p-dioxins (PCDDs), ten polychlorinated dibenzofurans (PCDFs), and twelve co-planar PCBs.
Twelve snapping turtle and five wood duck tissue samples were tested for seven polychlorinated dibenzo-
p-dioxins (PCDDs), ten polychlorinated dibenzofurans (PCDFs), and twelve co-planar PCBs.

For analysis of the PCDDs, PCDFs,  and co-planar PCBs, a Waters Autospec HRMS (High Resolution
Mass Spectrometer) coupled to an Agilent 6890 gas chromatograph equipped with a split/splitters
injector was used in lock mass correcting mode at 10,000 ppm resolution, using perfluorokerosene (PFK)
as the reference for mass calibration. An Agilent DBS-MS capillary column (60 m, id 0.320 mm,  0.250
|im film thickness J&W, USA) was used at a constant flow rate of 1.5 ml/min helium, using a splitless
injection of 275°C for the separation of both the PCDD/PCDF and PCB isomers.  Two oven programs
were used for two separate analyses: (1) PCBs: [Hold @ 130°C for 1  minute; then ramp to  235°C at 5°C
/minute; hold for 15 minutes; then ramp to 290°C at 10°C /minute; hold for 5 minutes]; (2)
PCDDs/PCDFs: [Hold @ 130°C for 1 minute; then ramp to 235°C at 5°C /minute; hold for ISminutes;
ramp to 290°C at 6°C /minute; hold for 12 minutes].  The electron energy for the HRMS used was
approximately 35 eV and the rest of the mass spectral lenses were tuned for maximum sensitivity.  The
HRMS was operated in SIM (single  ion monitoring) mode with mass ions and windows monitored
equivalent to a modified Method 1613  (Winters et al., 1996; Ferrario et al., 1997).  Surrogate recovery
standards, either a 13C PCB or 13C TCDD, were introduced in the initial extraction step in an amount,
depending on sample  amount, to equal a final  injection volume concentration of between 5-20 pg/|iL,
depending on the target analyte. The 13C labeled  surrogate recovery standards were used to  calculate the
recovery of the 13C labeled analogs, relative to a  13C labeled injection  standard (added to cleaned up
sample extract prior to MS analyses) and, to quantify native analyte concentrations adjusted for recovery.
A five or six point linear calibration  curve was used for the analyses ranging from  100 fg/|iL (2, 3, 7, 8-
TCDD) to levels as high as 1000 pg/|iL (PCB 118).

Mutagenicity Assays
For this study, the Salmonella mutagenicity assay was used to screen surface water, sediment, and
drinking water for mutagenicity. The Salmonella mutagenicity assay has been used extensively to
identify genotoxic substances in environmental samples (Claxton, 1985; Claxton et al., 1998, 2004;
Claxton and George, 2002; Chen and White, 2004; Claxton and Woodall, 2007; Claxton et al., 2010;
Maertens et al., 2004; Ohe et al., 2004; Richardson et al., 2007; Zwiener et al., 2007).  The assay is useful
in the present context because of its ability to identify mutagenic activity in surface waters (Ohe et al.,
2003, 2004), sediments (Chen and White, 2004), and drinking waters (Richardson  et al., 2007). The
assay determines the mutagenicity, and potential  carcinogenicity, of compounds and complex mixtures
(Mortelmans and Zeiger, 2000). However, because many carcinogens act by mutagenic mechanisms,
most organic carcinogens that are mutagens present a positive indication in the Salmonella assay.
Conversely, the assay has identified  some mutagens that have not been shown to be carcinogens.  The
Salmonella  assay is the most widely used genotoxicity assay for identifying environmental carcinogens
and for comparing locations, identifying sources, and identifying the likely carcinogens in complex
environmental mixtures (Claxton, 1997; Claxton et al.,  1998, 2010; MacGregor, 1994).

The Salmonella mutagenicity assays enabled the  assessment of complex, organic mixtures of air, soil,
and water by evaluation of organic extracts of these media. The results from such  analyses  provide an
integrated measure of the mutagenic activity and, thus, potential carcinogenic activity, of the organics in
   Page | 46                                                                     201

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environmental media.  In the present study we used the Salmonella mutagenicity assay to evaluate the
mutagenicity of river water, sediment from the river, and drinking water associated with the PIN.

To assess mutagenicity of Penobscot River water, drinking water, and sediment the sample extracts in the
Salmonella mutagenicity assay were tested with and without metabolic activation (Aroclor 1254-induced
Sprague-Dawley rat-liver S9, Moltox Inc., Boone, NC) following the procedures of Maron and Ames
(1983) with modifications from Claxton et al. (1987). The frameshift strain TA98 and the base-
substitution strain TA100 were used, which were provided by Dr. B.N. Ames, Children's Hospital
Oakland Research Institute, Oakland, CA. Strain YG1041 (derived from TA98) and strain YG1042
(derived from TA100) were also used, which over-express acetyltransferase and nitroreductase,
enhancing the sensitivity of the strains to aromatic amines and nitroarenes (Hagiwara et al.,  1993). These
YG strains were kindly provided by Dr. T. Nohmi, National Institute of Health Sciences, Tokyo, Japan.

The strains and metabolic activation condition (with or without S9) used for screening the samples were
chosen based on their particular sensitivities, their successful use in previous studies, and possible
anthropogenic sources of contamination.  Strain TA100 -S9, which detects direct-acting  mutagens that
induce base substitutions in deoxyribonucleic acid (DNA), has been used frequently when testing chlorinated
drinking water.  Strain TA98 detects agents that induce frameshift mutations and has been used with river
water and  sediments (Chen and White, 2004; Ohe et al., 2003, 2004). Surface water samples were tested
with YG1041 and YG1042, which express elevated levels of both nitroreductase and acetyltransferase
activity  and are used for the detection of mutagenicity in environmental samples when nitroarenes and
aromatic amines may be present.

Due to limited number of samples, each sample was first tested in the plate-incorporation assay at one
plate per dose (5-7 doses) and then repeated if the volume of extract available permitted.  The plates were
incubated for 72 h at 37°C and colonies counted with an AccuCount™ 1000 automatic colony counter
(Biologies, Inc., Manassas, VA).  The data were entered in the GeneTox Manager statistical analysis
program (Claxton et al., 1995) for mutagenic potency determination using the Bernstein  method
(Bernstein et al., 1982). The mutagenic potencies were calculated as revertants (rev) per  liter-equivalent
(L-eq) for the river and drinking waters and as rev per gram-equivalent (g-eq) for the sediment samples.
A positive result was defined as one in which the extract produced a dose-related increase of at least
twofold over the DMSO control number of revertants/plate; the DMSO  controls were used in the potency
calculations from the dose-response curves.

River-water samples were tested for mutagenicity in strains YG1041 and YG1042 with and without S9
metabolic  activation.  The first experiments were performed with eight doses (10-500 ml-eq/plate) using
YG1041.  A repeat test was performed using a dose range of 100-500 ml-eq/plate with YG1041; a single
experiment was performed with YG1042 with this dose range (with and without S9) due to limited
sample.

The first experiment was performed with the drinking-water samples in strains TA98 and TA100 without
S9 using the same doses used for the river-water samples; some of the lower doses for the repeat
experiments were not used.  A final  experiment was performed in TA100 using a dose range of 300-1000
ml-eq/plate without S9. River-sediment samples were tested in strains TA98, TA100, YG1041, and
YG1042 with and without S9. The first experiments were performed using a dose range of 0.2-10 g-
eq/plate, and repeat experiments were performed using a dose range of 0.1-1 g-eq/plate.
   Page | 47                                                                      201

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                                    Quality Assurance
US EPA Thomas Hughes conducting a TSA with USGS Charlie Culbertson and PIN DNR Dan Kusnierz at PIN
DNR office.

The Research team that designed this study included an array of experts such as toxicologists,
hydrologists, risk assessors, environmental health specialists and experts in Dioxin/Furan and PCB
analysis and Mercury analysis. (See Appendix D for list of experts). Collaborating with numerous
scientists and ATSDR assured the scope and procedures identified for this project met the objectives of
the PIN and that the methodologies employed are accepted within the scientific community.

To ensure the highest quality and reliability, a process of internal and external peer review by both cultural
and scientific experts was followed. The approach for this research project combined some of the elements of
consumption surveys such as interviewing Tribal  elders to determine recent natural resource utilization
patterns with careful identification of Tribal exposure factors (contaminant concentrations, pathways of
exposure).  The process used was culturally sensitive, respectful, drew on traditional environmental
knowledge (such as the observational expertise of elders), and was developed in partnership with tribal
cultural and technical experts.   The study was developed through a community-based participatory process,
which provided an avenue to foster a strong, communicative relationship. The PIN Natural Resources
Program's facilitation of this study with the PIN Tribal Community provided this assurance.

Thomas Hughes, EPA-ORD QA Manager, was the RARE Program QA Manager (PQAM). He assisted the
team by ensuring that the study complied with the QAPP and EPA policies and procedures with the
assistance of Steve  DiMattei from the EPA Regionl Chelmsford Laboratory.  The quality assurance (QA)
activities on this project were extensive due to the importance of this research program to the EPA and the
Penobscot Indian Nation (PIN).

A QA Statement, which is a listing of the five audits conducted during this RARE study, is in Appendix
E. An initial site visit of the Penobscot River near Indian Island in October 2007 was followed by
technical systems audits (TSA) in Old Town, ME and North Chelmsford, MA in 2008. A final data audit
was conducted in Old Town, ME in October 2010. A TSA was conducted by Barbara Collins, a QA
Manager at the EPA in RTF, in 2009 on the Salmonella (Ames)  mutagenicity testing of the river and
drinking water and  the river sediments for potential  carcinogenic potential. These audits demonstrated
   Page | 48
Final RARE Report August 2015

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that the procedures and data generated under this RARE program were scientifically acceptable.
Accurate shipping records were kept on fish, water, and sediment samples.  Information on exact
sampling site positions was recorded.

The Co-Pi on this RARE study at the EPA, Janet Diliberto, and the EPA PQAM, Thomas Hughes, were
involved in the review and helped to generate the RARE QA Project Plan (QAPP) through monthly
teleconferences with the Co-Pi in Boston (Valerie Marshall), the Senior Regional Scientist for Region 1
(Robert Hillger), and the entire study research team of over 20 scientists. The generation and approval of
this complex QAPP required an effort of the entire team and took a year to complete. After the generation
of the QAPP, teleconferences were held consistently and on an as needed basis throughout the duration of
this study.  To ensure that the data from this study was of the highest quality, EPA QA Officers Tom
Hughes and Steve DiMattei, EPA Region  1 laboratory,  reviewed each of following procedures: sampling
collection, sample handling, sample chemical analysis,  and laboratory QA/QC procedures. The quality of
the data from this study is considered exceptional, as verified by the review of the QAPP and SOPs, site
visits, and five QA audits.

                                         Peer Review
To ensure the reliability, credibility and integrity of this report, a formal EPA Peer Review of both the cultural
and scientific aspects of this report was conducted. The data collected from this study was accepted by all the
Agencies involved  in this RARE Project, i.e. US EPA, USGS, ATSDR, and US F&WS. Collaborating with
numerous scientists and ATSDR assured the scope and procedures identified for this project met the objectives
of the PIN and that the methodologies employed  are  accepted within the scientific community.  The
methodology used to reach the conclusions of this study incorporated information from a variety of disciplines,
including cultural and traditional environmental knowledge and ensured that the methodologies employed were
accepted within  the scientific community.  A number of scientific peer reviewed publications have been
developed based on the results of data collected from the study.  An independent peer review of the papers to
be published, including a Public Health Assessment by ATSDR was completed.

Tribal Risk Assessors were included as peer reviewers of this report, especially since the approach for this
research project is culturally sensitive and drew on traditional environmental knowledge (such as the
observational expertise of Tribal elders). The approach  for this study was developed in partnership with the
Penobscot Indian Nation. The PIN Natural Resource Department coordinated with the PIN  Tribal
Community and provided information pertaining to the sustenance practices of the Penobscot Indian Nation
tribal members.

The report was peer reviewed by a panel of experts according to the US EPA's Peer Review protocols. A
Confidential Draft Report was issued to a panel of 10 peer  reviewers that consisted of the following areas of
expertise:
 >  2 Tribal Risk Assessors;
 >  State Health Assessor;
 >  Mutagenicity Expert;
 >  Green Chemist;
 >  State Toxicologist (Maine);
 >  Research Chemist;
 >  Research Hydrologist; and,
 >  2 EPA  Toxicologists.
   Page | 49                                            Final RARE Report August 2015

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General and specific charge questions were developed by the RARE Research Team. A list of objectives was
used for focusing the peer review charge questions to obtain input from qualified colleagues ("the relevant
scientific community").  The Research Team worked with the Region 1 Peer Review Coordinator to assure
that the study was in compliance with the Agency's protocols.

All comments received from the peer reviewers were evaluated and addressed. All significant
recommendations were incorporated into the report. Once all recommendations from the peer reviewers were
incorporated into this report, the team provided an overview of the study to the PIN tribal council and
community. The RARE team shared the draft confidential report and all other publications to gather input
from the Tribal Leaders and the tribal community prior to finalizing the report. Any significant comments
submitted by the Tribal Community are incorporated  into the final report.

                                      Data Validation
Data review, validation, and verification are the processes for documenting the degree to which the
project objectives were met, individually and collectively, and to estimate the effect of any QA/QC
procedural deviations on the ability to use the data.

EPA Region 1 (EPA-New England) has three tiers of data validation (DV):
•  Tier I - The analytical laboratory data package is checked for completeness and any Performance
   Evaluation (PE) samples are checked for accuracy;
•  Tier II - The quality control (QC) results are checked against acceptance criteria. Based on the QC
   results, reported laboratory data are qualified as either acceptable, estimated (J) or rejected (R);
•  Tier III - An in-depth examination of instrument-generated analytical data is performed to ensure the
   accuracy of the results reported. The calculation of reported results is  verified. Tier III is the
   preferred level of validation for human health and ecological risk assessments.

Tier I validation - determines whether or not the laboratory provided the contract or agreement required
deliverables. This is called a completeness check.  A Tier I validation includes the evaluation of
Performance Evaluation (PE) sample results which demonstrate laboratory performance at the time of
field sample analysis. Depending on the PE sample results, field sample data may or may not be
qualified as acceptable, estimated (J) or rejected (R).  A Tier I data validation report documents missing
data/information that could not be retrieved from the  laboratory, a discussion of the PE sample  results,
and a summary table of the laboratory results (unqualified).

Tier II validation - includes a Tier I review and the QC sample results are reviewed.  Data qualifiers are
applied to the laboratory results based on the PE and QC sample results and the project objectives. The
results of a Tier II validation are documented on worksheets specific to parameters reviewed. The report
includes a narrative discussion for each parameter reviewed and a data summary table which documents
the qualified data.

Tier III data validation - includes Tier I and Tier II data validation procedures and a Tier III review,
which includes in-depth qualitative and quantitative determination of accuracy. This requires re-
calculating results for instrument generated reports and an examination of the various instrument outputs
which document the results reported.  During Tier III the gas chromatograms, the mass spectra and
instrument out-put are examined to ensure the data corroborate the reported results. The data are checked
for calculation, transcription and identification errors. Proper compound identifications are confirmed
   Page | 50                                             Final RARE Report August 2015

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and discrepancies resolved. The Tier III DV report is the same as the Tier II report however, there is
potential for additional qualification of data as a more in-depth review is performed.

The validation for this project was 90% at Tier II level and 10% at Tier III (Table 4). The table below
summarizes the data validation that was conducted for this study. EPA data validator, Steve Stodola
reviewed and interpreted the data validation. Each laboratory that performed analysis for this project
applied data validation flags in the form of Remark Codes to those sample results that fell outside of the
QC acceptance criteria, for data under its purview.
   Page | 51                                                                       201

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RARE Penobscot Data Validation
Analyte
D/F OPP ECL
PCBWHO
OPP ECL
PCB Cong
CERC USGS
Total Hg ESS
c/o CERC
MeHg
FGS
D/F OPP ECL
WHO PCB
OPP ECL
PCB Cong
CERC USGS
Total Hg
NERL EPA
MeHg
D/F OPP ECL
WHO PCB
OPP ECL
Total Hg
NERL EPA
MeHg
FGS
TOC
ESS c/o CERC
Grain Size
ESS c/o CERC
Media
Sediments
Sediments
Sediments
Sediment
Sediment
Fish Fillet
Fish Fillet
6 Fish,
6 Ferns,
4 Ducks,
2 Turtles
27 Fish,
2008
7 Fish,
2009
Fish Fillet
2 Turtles &
5 Ducks
2 Turtles &
5 Ducks
2 Turtles,
5 Ducks,
7 Ferns
2 Turtle,
5 Duck,
7 Plants
Sediment
Sediment
No. of
samples
for Tier
II
19
19
8
21
21
30
30
6
NA
NA
6
6
NA
14
21
0
No. of
Samples
for Tier
III
2
2
1
0
0
4
4
1
NA
NA
1
1
NA
0
0
0
        Table 4: Data validation for Penobscot data in the RARE study.
Page |  52
Final RARE Report August 2015

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Analytical Results from Reaches 1-6: Contaminants Concentrations
The objective of the preliminary risk assessment was to assess the level of contaminants that the
Penobscot tribal members are potentially exposed to from partaking in their tribal cultural practices by
comparing maximum concentrations of detected chemicals in a medium with conservative health risk-
based concentrations. The intent of the preliminary risk assessment was to identify which Penobscot
River reaches, and what types of exposures to which types of contaminants, are found to be above the
Project Quantification Limits or risk-based concentrations identified for the chemicals analyzed in this
study.

The targeted contaminants analyzed included PCDDs, PCDFs, WHO-coplanar PCBs, total-PCBs, total
mercury, and methyl mercury. Concentrations of selected chemical contaminants were measured  in
samples offish and streambed sediment collected from six reaches of the Penobscot River. Fish
(including eel, pickerel, perch, smallmouth bass, and bullhead) and sediment (from wading areas) were
collected. Skinless fillets were analyzed. The goal was to collect and analyze one composite sample of
each species offish from each reach. In addition, wood duck muscle, snapping turtle muscle, and plants
were collected and analyzed. The contaminant concentrations for biota and sediment are provided in
Tables 5-10. The maximum contaminant concentrations by sample type are presented in Table  11, and
by reach in Table 12.
   Page | 53                                            Final RARE Report August 2015

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Contaminant concentrations in sediment and biota in Reach 1, Milford Dam Impoundment
(MIL)

SAMPLES
(A, B,C,D)

Sediment A
(SED)
BlmP
C
D
Average
Chain
Pickerel A
(CP)
Yellow
Perch
(YP)
White
Perch A
(WP)
Smallmouth
Bass A
(SMB)
Brown
Bullhead A
(BBH)
American
Eel A
(EEL)
Wood duck
(WODU)
Fiddlehead
Ostrich A
Fern (OSF)
Medicinal
Plant
(MP)
Snapping
Turtle A
(SNTU)
Dioxins/Furans
(17 Congeners)
Concentration
TEQ pg/g
20. 9a

21. 1a
11. 4a
10. Oa
15.9a
0.021 7b





0.1 77b


0.0423b


0.220 b


1.34b


0.111b

0. 000321 b




3.51 b

WHO-PCBs
(12 Congeners)
Concentration
TEQ pg/g
0.285a

0.21 9a
0.1 50a
0.191a
0.21 1a
0.0207b





0.206b


0.0576b


0.1 17b


1.1 6b


0.1 76b

NDb
-------
Contaminant concentrations in sediment and biota in Reach 2, Sunkhaze-West Enfield Dam
(SWE)
SAMPLES
(A,B,C)
A
Sediment Q
(SED)
C
Average
Chain ^
Pickerel d
(CP) A
Yellow
Perch A
(YP)
White
Perch
(WP)
Smallmouth
Bass A
(SMB)
Brown
Bullhead A
(BBH)
American
Eel A
(EEL)
Wood duck
(WODU) A
Fiddlehead A
Ostrich Ad
Fern (OSF)
Medicinal
Plant
(MP)
Snapping
Turtle
(SNTU)
Dioxins/Furans
(17 Congeners)
Concentration
TEQ pg/g
16.3a
13.8a
4.33a
11. 5a
0.0232b
0.0206b

0.0561 b
0.0872b
0.646b
0.171b
0.0001 79b
NDb>d>e


WHO-PCBs
(12 Congeners)
Concentration
TEQ pg/g
0.373a
0.289a
0.1 67a
0.276a
0.0361 b
0.0151b

0.111b
0.0631 b
0.533b
0.255b
NDb>e
NDbd
0.377b

0.945b
0.423b
0.666b
0.032b
NDa>e
NDa,d,e



Table 6: Contaminant concentrations in sediment and biota in Reach 2, Sunkhaze-West Enfield Dam (SWE).
 a Dry Weight
 b Wet Weight
 c Total:  Total concentration of congeners in this class of contaminants
 d Duplicate Sample
 e ND: Non-Detect
 lmp Impoundment
     Page | 55
Final RARE Report August 2015

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Contaminant concentrations in sediment and biota in Reach 3, West Enfield Dam
Impoundment (WEI)


SAMPLES
(A, B,C,D)
A
Sediment
(SED) Blmp
C

D
Average
Chain
Pickerel A
(CP)
Yellow
Perch A
(YP)
White
Perch
(WP)
Smallmouth
Bass A
(SMB)
Brown
Bullhead A
(BBH)
American
Eel (EEL) A
Wood A
Duck d
(WODU) A
Fiddlehead A
Ostrich
Fern (OSF)
Medicinal
Plant A
(MP)
Snapping
Turtle A
(SNTU)
Dioxins/Furans
(17 Congeners)

Concentration
TEQ pg/g
5.89a
29. 8a
26. 6a

20. 5a
20. 7 a
0.0185b


0.0372b





0.0443b


0.1 89b


1.18b
0.1 37b
0.178be


0.1 98b

Total TEQ
(29
Congeners)
Concentration
TEQ pg/g
6.13a
30. 4a
26. 9a
20. 9a

21. 1a
0.0399b


0.0641b





0.1 32b


0.297b


2.23b
0.287b
0.454b
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Contaminant concentrations in sediment and biota in Reach 4, Mohawk-Mattaseunk Dam
(MM)
SAMPLES
(A,B,C)
A
Sediment B
(SED)
C
Average
Chain Pickerel A
(CP) A
Yellow Perch .
(YP)
White Perch .
(WP) M
Small-Mouth ^
Bass d
(SMB) A
Brown Bullheac A
(BBH) A
American Eel .
(EEL)
Wood duck A
(WODU) A
A
Fiddlehead e
Ostrich Fern
(OSF) B
Medicinal
Plant A
(MP)
Snapping .g
Turtle h
(SNTU) B
Dioxins/Furans
(17 Congeners)
Concentration
TEQ pg/g
11. 5a
26.0a
4.43a
14.0a
0.0151b
0.01 39b
0.1 56b
0.0635b
0.0597bf
0.51 3b<9
WHO-PCBs
(12
Congeners)
Concentration
TEQ pg/g
0.389a
1.19a
0.1 88a
0.589a
0.0227b
0.0204b
0.246b
0.1 57b
0.184bf
1.18b<9
Total TEQ
(29
Congeners)
Concentration
TEQ pg/g
11. 9a
27. 2a
4.62a
14.6a
0.0378b
0.0343b
0.402b
0.221 b
0.244bf
1.69b'9
Total PCBs
142 Congeners)
Concentration
ng/g
44.4a
44. 4a



0.432b
1.25b'd


4.052b
0.322b
0.224b

21 .4b
Methyl
Mercury
Concentration
ng/g
1.43a
0.798a
2.63a
1.62a






16.8b
6.3a
1.3a
NDb
605b
202b
Mercury
Concentration
M9/9
0.093a
0.1 50a
0.066a
0.1 03a
0.31 6b
0.1 46b
0.467b
0.965b 0.887b
0.71 3b'd
0.1 80b
0.337b
0.026b
0.00744a
NDa'e
0.00861 b
0.577b
0.222b
Table 8: Contaminant concentrations in sediment and biota in Reach 4, Mohawk-Mattaseunk Dam (MM)
 a Dry Weight                                                 d  Duplicate Sample              9 Year 2009
 b Wet Weight                                                 e  ND: Non-Detect
 c Total:  Total concentration of congeners in this class of contaminants        f  Year 2008
     Page | 57
Final RARE Report August 2015

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Contaminant concentrations in sediment and biota in Reach 5, Mattaseunk Dam
Impoundment (MAT)


SAMPLES

(A, B,C,D)
A'mP

(SED) B
C
D
Average
Chain Pickerel A
(CP) A
Yellow Perch
(YP)
White Perch A
(WP) Ad
Smallmouth
Bass A
(SMB)
Brown Bullheai .
(BBH)
American
Eel A
(EEL)
Wood duck
(WODU)
Fiddlehead
Ostrich Fern
(OSF)
Medicinal
Plant A
(MP)
Snapping
Turtle A
(SNTU)
Dioxins/Furans
(17 Congeners)

Concentration
TEQ pg/g
19.9a

54. 8a
94. 9a
93. 8a
65. 9a
0.0579b


0.495b
0.531 bc

0.727b

5.45b







0.0240b


2.80b

Total PCBs
(142
Congeners)
Concentration
ng/g
76.7a

168a


122a





1.10b
















Methyl Mercury


Concentration
ng/g
1.15a

5.28a
3.65a
8.98a
4.77a

















NDb


938b

Mercury


Concentration
M9/9
0.24a

0.56a
0.64a
3.48a
1.23a
0.588b


0.627b
0.545b
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Contaminant concentrations in sediment and biota in Reach 6, Control Reach- East Branch-
Salmon Stream Lake (EBS)
SAMPLES
(A,B,C)
A
Sediment B
(SED)
C
Average
Chain Pickerel A
(CP) A
Yellow Perch .
(YP)
White Perch .
(WP) A
Smallmouth
Bass A
(SMB)
Brown Bullhead .
(BBH) M
American Eel A
(EEL) Ad
Wood duck
(WODU)
Fiddlehead
Ostrich Fern A
(OSF)
Medicinal Plant A
(MP) Ad
Snapping Af
Turtle
(SNTU) B 9
Dioxins/Furans
(17 Congeners)
Concentration
TEQ pg/g
0.651 a
0.07603
0.1 48a
0.292a
0.0161b
0.00370b
0.1 46b
0.0428b
0.1 07b
0.1 78b

NDb>e
NDbe
0.0360b
NDb,d,e
0.0876be
0.0360b
NDb,d,e
0.109bd

NDa>e
0.00289b
0.00292b'd
0.21 5b
-------
Summary of Contaminant Concentration Data
Below is a summary of the contaminant concentration data for sediment and biota in the six study reaches, as
presented in Tables 5-10:

Sediments
   The concentrations of PCDD/PCDF and co-planar PCBs (pg/g, TEQ dry weight) were similar in
   sediments from Reaches 1-4, but differed markedly from those in sediments of Reach 5.  Sediments
   from Reach 5 had the highest single and average concentrations of PCDDs/PCDFs and co-planar
   PCBs of sediments from any reach and were more than 200- and 15- fold higher for PCDD/PCDF
   and co-planar PCBs, respectively, than those in sediments from Reach 6, the control site.  Sediment
   concentrations  of these compounds in Reaches 1-5 all differed markedly from those in Reach 6.

Animals
   With respect to the concentrations of PCDDs/PCDFs and co-planar PCBs in the tissue (pg/g, TEQ
   wet weight), there  appear to be similarities and differences by species and locations.
   1) For the chain pickerel (a water column predator at the top of the food web species) with respect to
      the concentrations of PCDDs/PCDFs in the fillet tissues, there appears to be a similarity between
      Reaches  1-4 and Reach 6, the control reach, but a marked difference between those five reaches
      and Reach  5 (Mattaseunk Dam/Impoundment). There was no difference among any of the reaches
      in the concentration of co-planar PCBs in chain pickerel.
   2) For the yellow perch, a water column predator, with respect to the concentrations of
      PCDDs/PCDFs in the fillet tissues, there appears to be  a difference between Reaches 1-4 and
      Reach 6, but no difference among any of the reaches in the concentration of the co-planar PCBs in
      yellow perch. No yellow perch were collected in Reach 5.
   3) For the white perch, another water column predator, with respect to the concentrations of
      PCDDs/PCDFs in the fillet tissues, there appears to be  a similarity between Reaches 1-4 and
      Reach 6 and a marked difference between those five reaches and Reach 5. There was no
      difference among any of the reaches in the concentration of co-planar PCBs in white perch.
   4) For the small mouth bass, another water column predator, with respect to the concentrations of
      PCDDs/PCDFs and co-planar PCBs in the fillet tissues, there appear to be no differences among
      any reaches.
   5) For the brown bullhead catfish,  an opportunistic bottom feeder, with respect to the concentrations
      of PCDDs/PCDFs in the fillet tissues, there appears to be a similarity between Reaches 1-4 and
      Reach 6 and a marked difference between those five reaches and Reach 5. There was no
      difference among any of the reaches in the concentration of co-planar PCBs in brown bullhead
      catfish.
   6) For the freshwater eels, a bottom dwelling predator, with respect to the concentrations of
      PCDDs/PCDFs in the fillet tissues, there appears to be  a difference between Reaches 1-4 and
      Reach 6 and a marked difference between those five reaches and Reach 5. With respect to the
      concentrations of co-planar PCBs, there appear to be marked differences between Reaches 1-4
      and Reach  6, and between Reach 5 and Reach 6.
   7) For the snapping turtle, an opportunistic carnivore/scavenger, with respect to the concentrations of
      PCDDs/PCDFs and the co-planar PCBs in the fillet tissues, there appears to be a difference
      between Reaches 1-4 and Reach 6 and a marked difference between Reach 5 and Reach 6.

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       8)  For the wood ducks, there were no differences in concentrations of PCDDs/PCDFs and co-planar
          PCBs among the four reaches.  No wood ducks were collected in Reaches 5 and 6.

       It appears that the concentrations of PCDDs/PCDFs and co-planar PCBs in some biota reflect their
       relative concentrations in the sediments, with the highest concentrations in biota from Reach 5.
       Those aquatic organisms that are in direct contact with the sediments appear to have the highest
       concentrations of PCDDs/PCDFs and co-planar PCBs (i.e., brown bullheads, eels, and snapping
       turtles).

       Plants
       The plants contained no appreciable concentrations of any of the pollutants.

   Tables 11 to 12 provide a  comprehensive overview of the highest measured contaminant concentration for all
   flora, fauna and sediments sampled per reach. An analysis of the risk associated with these contaminant
   concentrations can be found in the Exposure Assessment Section of this report.
Highest Contaminant Concentrations in Sediment and Biota by Reach

Total
Dioxin/Furan (17
Congeners)
Total WHO-PCB
(12 Congeners)
Total TEQs (29
Congeners)
Total (142) PCB
Congeners
Methyl Mercury
Mercury
Reach 1
Turtle
(3.51pg/g)
Turtle
(1.35pg/g)
Turtle
(4.86 pg/g)



Reach 2






Reach 3
Plants
(0.064 pg/g)




Smallmouth
Bass
(0.979 jig/g)
Reach 4
Duck
(0.426 pg/g)


Smallmouth Bass
(1.25ng/g),
Turtle
(21.4ng/g)


Reach 5
Sediment (65.9 avg. pg/g)
Eel (4.02 pg/g)
Brown Bullhead (0.534 pg/g)
Sediment (1.75 avg. pg/g)
Eel (1.43 pg/g)
White Perch (0.3 11 pg/g)
Sediment (67.8 avg. pg/g)
Eel (5.45 pg/g)
White Perch (0.812 pg/g)

Sediment (4.77 avg. ng/g)
Turtle (938 ng/g),
Sediment (1.23 avg. (ig/g)
Eel ( 0.739 jig/g)
Turtle 1.046jig/g)
Table 11. Highest contaminant concentrations in sediment (dry weight) and biota (wet weight) by study Reach.
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Highest Contaminant Concentrations found in Sediment and Biota by Sample Type
Sample Type
Sediment
(Avg.)
Eel
Fish (other
than Eel)
Duck
Turtle
Plants
Total
Dioxin/Furans
(17
Congeners)
Concentration,
Dioxin Toxic
Equivalent
(TEQ pg/g)
65.9
4.02
0.534 (Brown
Bullhead)
0.426
3.51
0.064
Total WHO-
PCB
Congeners
(Dioxin-like
PCBs; 12
congeners)
Concentration,
(TEQ pg/g)
1.75
1.43
0.3 11 (White
Perch)
0.652
1.35
0.0402
Total TEQs
(29
Congeners)
Concentration
TEQ Pg/g
67.8
5.45
0.812
(White Perch)
1.08
4.86
0.0902
Total PCB
(142
Congeners)
Concentration
(ng/g)
122

1.25
(Smallmouth
Bass)
5.01
21.4
1.15
Methyl
Mercury
Concentration
(ng/g)
4.77
No samples
tested for
Methyl
Mercury
No samples
tested for
Methyl
Mercury
47.9
938
6.3
Mercury
Concentration
(^g/g)
1.23
0.739
0.979
(Smallmouth
Bass)
0.049
1.046
0.00861 (ig/kg
Table 12. Highest contaminant concentrations found in sediment (dry weight) and biota (wet weight) by sample type.
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       Exposure Assessment ofPenobscot Indian Nation tribal members

Risk Assessment vs. Risk Management
Risk assessment is the process of calculating the exposure dose for particular exposure pathways and then
calculating the risk of cancer and non-cancer health effects. Risk assessment is the use of a factual base to
define the health effects of exposure to individuals or population to hazardous materials or situations.
Selection of maximum acceptable risk is a policy or risk management decision, rather than a risk
assessment calculation. The risk assessment calculations provide an estimate of the likely quantitative
level of risk using the best available exposure and toxicity information.

Risk management is the process of deciding what to do about the risks that were calculated in a risk
assessment. For most Federal Agencies, risk management is the process of weighing policy alternatives
and selecting the most appropriate regulatory action, integrating the results with engineering data and with
social, economic, and political concerns to reach a decision. For stakeholders, risk management is a
decision of how much risk is acceptable. For this RARE study, EPA selected risk management criteria
consistent with EPA criteria for water quality standards and other EPA environmental programs.  (See
Table 17)

EPA Exposure Assessment
An exposure assessment is the determination or estimation (quantitative or qualitative) of the magnitude,
frequency, duration and route of exposure. The exposure assessment is a three step process consisting of
characterizing the exposure setting, identifying the exposure pathways and quantifying the exposure.

EPA uses the following equation for estimating the exposure to contaminants:

Site Dose = Ingestion Rate x Exposure Frequency x Exposure Duration
                          Body Weight x Averaging Time

For this RARE study, the team was able to use  tribal ingestion rates. In 2009, the USEPA and the
federally recognized Maine Tribal Nations worked in a collaborative effort to develop exposure scenarios
that reflect the Maine tribal traditional cultural uses of natural resources, i.e. The Wabanaki Traditional
Cultural Lifeways Exposure Scenario. The Wabanaki Traditional Cultural Lifeways Exposure Scenario
was developed by gathering information from several types of literature (ethnohistorical, ecological,
nutritional, archaeological, and biomedical) to develop a description of Wabanaki traditional subsistence
lifestyles and diets through the lens of natural resource use and activities necessary to survive and thrive in
Maine environments. Although the information used to develop a nutritionally complete diet is taken
from literature that describes diets from the 16th, 17th, 18th, and 19th centuries, this information is still
relevant today  even if that diet is eaten by fewer people at present.

The Wabanaki Traditional Cultural Lifeways Exposure Scenario (Wabanaki Exposure Scenario) describes
the lifestyle that was universal when resources were in better condition and that some Tribal Members still
practice today. The Wabanaki Exposure Scenario reflects full traditional resource uses. Therefore, rather than
using  nationwide conservative default consumption rates to assess potential exposure, realistic tribal
consumption rates combined with data collected for contaminants in water, soil/sediment, flora or fauna were
used to determine realistic potential  exposures to Tribal Members of the Penobscot Indian Nation. By
coupling contamination information gathered through the RARE study with the ingestion factors developed
in the Wabanaki Exposure Scenario, the RARE team was able to assess the level of exposure to Penobscot
Indian Nation Tribal Members that occurs when they sustain their traditional life ways. Accordingly, this


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 study reflects EPA's National Tribal Science Council's Tribal Health and Well Being paradigm by
 incorporating tribal culture to assess exposure risks to tribal health and the ecosystem.

 The exposure assessment for the Penobscot River study assumed that the diet was the "Inland Non-
 Anadromous" diet presented in the Wabanaki Exposure Scenario. Below is a table that shows the tribal
 consumption/ingestion rates for the flora and fauna analyzed in this RARE study based on the Wabanaki
 Exposure Scenario. The team chose to use the inland non-anadromous diet described in the Wabanaki
 Exposure Scenario because it appeared to be the diet most closely aligned with the Penobscot Indian
 Nation's cultural lifestyles.
Food Categories and Consumption Rates for Wabanaki Tribal Populations
Tested
Biota
Chain
Pickerel
Yellow
Perch
White
Perch
Smallmouth
Bass
Brown
Bullhead
American
Eel
Wood duck
Fiddlehead
Fern
Medicinal
Plant
Snapping
Turtle
Symbol
CP
YP
WP
SB
BB
AE
WD
FF
MP
ST
Food Category
Resident fish and other
aquatic resources
Resident fish and other
aquatic resources
Resident fish and other
aquatic resources
Resident fish and other
aquatic resources
Resident fish and other
aquatic resources
Anadromous and marine
fish and shellfish
Fowl and Eggs
Greens, Tea (includes
leaves, stems medicinal
plants)
Greens, Tea (includes
leaves, stems medicinal
plants)
Resident fish and other
aquatic resources
Consumption Rates (g/day)
Inland
Anadromous
114
114
114
114
114
400
70
133
133
114
Inland
Non-Anadromous
286
286
286
286
286
0 (286)1
70
133
133
286
Coastal
57
57
57
57
57
457
120
133
133
57
Table 13- Food Categories and Consumption Rates for Wabanaki Tribal Populations Source: Section 7.2
 Wabanaki Traditional Cultural Lifeways Exposure Scenario
1 Eel are catadromous rather than anadromous and are considered to be resident fish for much of their life cycle

 The Definition for the diets can be found in the Wabanaki Traditional Cultural Lifeways Exposure Scenario and
 are defined as follows:
 •     Inland Anadromous = inland communities living on rivers with anadromous fish runs.
 •     Inland Non-Anadromous = inland communities without access to anadromous fish runs.
 •     Coastal = communities living where coastal resources are available.

 As shown in Table 13 above, the inland non-anadromous diet consumption rates are 286 g/day for each
 freshwater fish species (including eel) and snapping turtle, 70 g/day for wood duck, and 133 g/day for both
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Fiddlehead Ostrich fern and medicinal plant. Although the American eel is catadromous (marine spawning)
rather than anadromous (freshwater spawning), it is appropriate to include eel in the Inland Non-Anadromous
diet at the same ingestion rate as resident fish because eel are available for long periods of time in the river
where PIN members fish.

The contaminant concentrations in each type of biota from the various river reaches are shown in Tables 5-
10 under the Analytical Results: Contaminant Concentrations section of this report. These data were
produced by the Office of Research and Development and the US EPA Environmental Chemistry
Laboratory, under the Office of Pesticide Programs (OPP). Tables 5-10  summarize the results of the
analysis of dioxins/furans, dioxin-like PCBs, total PCBs (as the sum of 142 PCB congeners), methyl
mercury, and total mercury. Although this table includes data for sediment, this exposure assessment only
evaluated ingestion of biota. The concentrations of dioxins/furans and PCBs in biota were reported as the
contaminant concentration per gram of wet weight tissue (i.e. on a wet weight basis).  The concentrations
of methyl mercury and total mercury were also reported as the concentration per gram of wet weight tissue,
but also on a dry weight basis for fiddlehead ostrich fern.  Dry weight concentrations in fern were not
adjusted to wet weight because percent moisture data were not readily available for the biota samples.
Since dry weight contaminant concentrations are always higher than wet weight contaminant
concentrations in the same sample, and ingestion rates are based on wet weight, the mercury risks of fern
ingestion are overestimated by an unknown amount (for example, the wet weight concentration would be
about five times lower than the dry weight concentration if the tissue is 80% moisture). This
overestimation is insignificant because, as shown later, the overestimated non-cancer hazard quotients of
mercury in fern were between 0.1  and 0.02.

Mercury was reported as total mercury for all the fish species. Both methyl mercury and total  mercury for
all fern and duck samples, and for two of four turtle samples was reported. Methyl mercury is an organic
form of mercury that is much more toxic than inorganic mercury. Therefore, for risk assessment
purposes, it is important to know how much of the mercury in food is in the form of the more  toxic
methyl mercury.  The percent methyl mercury in a sample is calculated as the concentration of methyl
mercury divided by the concentration of total mercury (in the same concentration units), multiplied by
100.  In higher trophic level carnivorous fish, the ratio of methyl mercury to total mercury generally
approaches unity, meaning that almost all of the mercury in the fish fillet is in the methyl mercury form.
(See, Wiener et al., 2003)  It is generally assumed that >90% of mercury in higher trophic level
carnivorous fish is in the methyl mercury form, although site-specific variables and trophic level can
influence the ratio of methyl mercury to total mercury in fish.  The team  agreed that analyzing the
Penobscot fish fillet samples for total-mercury was a cost effective, accurate, and a slightly conservative
way to estimate the level of methyl mercury.

The percent methyl mercury in one fern sample (Reach  4) was 85%, but could not be calculated in four
other samples because methyl mercury was measurable but total mercury was lower than the detection
limit.  The percent methyl mercury in wood duck from five paired samples (i.e. both methyl mercury and
total mercury detected in the same sample) was 98%, 83%, >100%, 14 %, and 65% (average = 72%).
The percent methyl mercury in turtle from two paired samples was greater than 100% in each sample.
This impossible result is an artifact of variability in the analytical methods, but indicates that,  as in fish,
almost all of the mercury in snapping turtle is in the form of methyl mercury. Since it can be assumed
that 100% of the mercury in fish is methyl mercury, and most of the mercury in duck, turtle, and fern is
shown to be in the form of methyl mercury, it was assumed that all mercury concentrations used in the
risk screening were in the form of the more toxic methyl mercury.
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Although there was often more than one sample result for a particular species/analyte/reach combination,
the maximum concentration was selected for preliminary risk assessment, rather than calculating average
concentrations. In almost all cases there were no more than two samples of the same
species/analyte/reach combination.  EPA risk assessment guidance indicates that the use of the maximum
concentration is appropriate when there are too few samples to calculate the 95% upper confidence limit
of the mean, which requires three or more samples, and preferably more.

Toxicity Assessment
In order to determine the adverse health risks to the Penobscot tribal members of the chemicals tested, a
toxicity assessment of the contaminants was conducted.  A toxicity assessment is the characterization of the
toxicological properties and effects of a substance, specifically the dose response relationship associated with
a particular route of exposure.  The basic objective of a toxicity assessment is to identify what adverse health
effects a chemical causes and how the appearance of these adverse effects depends on exposure level (dose).
The toxic effects of a chemical frequently depend on the route of exposure (oral, inhalation, dermal) and the
duration of exposure (subchronic, chronic, or lifetime). Thus, a full description of the toxic effects of a
chemical includes a listing of what adverse health effects the chemical may cause and how the occurrence of
these effects depends upon dose, route, and duration of exposure.

The toxicity assessment process is usually divided into two parts: the first characterizes and quantifies the
non-cancer effects of the chemical, while the second addresses the cancer effects of the chemical. This two-
part approach is employed because there are typically major differences in the time-course of action and the
shape of the dose-response curve for cancer and non-cancer effects, http ://www2.epa. gov/region8/human-
health-toxicity-assessment. For example, toxicity of non-carcinogens is expressed as Reference Dose, the
dose (e.g. mg contaminant/kg body weight per day) considered by EPA to have no adverse effects; while the
toxicity of carcinogens is expressed as Cancer Slope Factor, the cancer risk probability/unit dose (e.g. risk
probability per mg contaminant/kg body weight per day). Cancer risk (CR) of carcinogens is expressed as a
probability of getting cancer due only to the exposure at the area of interest, rather than from all causes. It is
calculated by multiplying the calculated lifetime average daily dose of the chemical at the area of interest by
the chemical's cancer potency, which is also called the Slope Factor. The Slope Factor is derived by EPA,
preferably, or by other agencies based on data from the scientific literature.  Cancer risks are expressed as an
incremental lifetime cancer risk (ILCR) as a probability such as 1-in-l million. This probability can also be
expressed as 1E-06 or IxlO"6. ILCR values of 1-in-l million and less are generally considered to be of "no
concern".  For this report ILCR was simplified to CR (cancer risk). The exposure assumptions and toxicity
values used for the risk assessment are provided below.
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Exposure Assumptions
Exposure
Duration
Exposure
Frequency
Lifetime
Body Weight
Ingestion Rates
30 years
350 days/year
70 years
70kg
70 g/day
(wood duck)
133 g/day
(plants)
286 g/day
(fish)
Toxicity Factors
Chemical

TCDD-TEQ
meHg
PCB
RfDo
(mg/kg-
day)
7.0E-10
l.OE-04
2.0E-05
Source
IRIS
IRIS
IRIS
CSF
(mg/kg-
day)-1
1.3E+05
NA
2.0E+00
Source
CALEPA

IRIS

 Table 14: Exposure assumptions and Toxicity factors
 RfDo = oral Reference Dose                CALEPA = California Environmental Protection Agency
 CSF = oral Cancer Slope Factor             NA = Not Available
 IRIS = EPA Integrated Risk Information System

 (Note: toxicity values for PCB are those for Aroclor 1254; the three ingestion rates are for various food items as
  identified in text.)

Twelve of the 209 possible forms (congeners) of PCBs have dioxin-like activity. Of the 209 possible PCBs,
142 were measured by the analytical method used in this  study. The cancer and non-cancer risks of the
twelve dioxin-like PCB congeners are evaluated separately from the other PCBs using the Reference Dose
and Slope Factor for dioxins/furans and a Toxic Equivalents Scheme described below:

Toxic Equivalents Scheme (TEFs & TEQs)
The chlorinated chemicals known as polychlorinated dibenzo-para-dioxins (PCDDs or dioxins),
polychlorinated dibenzofurans (PCDFs or furans) and PCBs occur as mixtures of congeners. There are 75
dioxin congeners, 135 furan congeners, and 209 PCB  congeners, each with its own toxic potency.  To
express the overall toxicity of a given mixture of these chemicals as a single number, the concept of Toxic
Equivalence is used (World Health Organization, 201  la, b).  The toxicities of dioxin, furan and PCB
congeners are expressed relative to the most toxic dioxin  congener (2, 3, 7, 8- tetrachlorodibenzo-p-dioxin,
TCDD), which has a reference toxicity of 1. Under this scheme, each congener is attributed a specific
"Toxic Equivalency Factor" (TEF), indicating the degree of its toxicity compared to 2, 3,  7, 8-TCDD. To
calculate the total TCDD toxic equivalent (TEQ) of a  dioxin/furan, PCB mixture, the concentration of each
toxic compound is multiplied by its Toxic Equivalency Factor (TEF) and then added together.

For this study we analyzed 17 congeners of dioxins/furans and 12 congeners  of PCBs [the World Health
Organization (WHO-PCBs)], for a total of 29 congeners that contribute to the TEQ.

The TEQ scheme refers only to adverse effects (e.g. cancer, non-cancer) associated with the interactions of
these chemicals with cellular aryl  hydrocarbon (Ah) receptors. Other toxic effects of dioxins and dioxin-like
compounds are not quantified by this method. TEF values vary for different animal species.

The following table contains the various dioxin-like toxicity equivalency factors for Dioxins, Furans and
PCBs (Van den Berg et al. 2006). which are the World Health Organization 2005 values.
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Dioxin Toxicity Equivalence Factors
Chlorinated
dibenzo-p-dioxins







Chlorinated
dibenzofurans










Dioxins and Furans
2,3,7,8-TCDD
,2,3,7,8-PeCDD
,2,3,4,7,8-HxCDD
,2,3,6,7,8-HxCDD
,2,3,7,8,9-HxCDD
,2,3,4,6,7,8-HpCDD
OCDD

2,3,7,8-TCDF
1,2,3,7,8-PeCDF
2,3,4,7,8-PeCDF
1,2,3,4,7,8-HxCDF
1,2,3,6,7,8-HxCDF
1,2,3,7,8,9-HxCDF
2,3,4,6,7,8-HxCDF
1,2,3,4,6,7,8-HpCDF
1,2,3,4,7,8,9-HpCDF
OCDF
TEF
1
1
0.1
0.1
0.1
0.01
0.0003

0.1
0.03
0.3
0.1
0.1
0.1
0.1
0.01
0.01
0.0003
PCBs

Non-or//zo
Mono-ortho
Di-ortho *
IUPA
CNo.
77
81
126
169
105
114
118
123
156
157
167
189
170
180
Structure
3,3',4,4'-TetraCB
3,4,4',5-TetraCB
3,3',4,4',5-PeCB
3,3',4,4',5,5'-HxCB
2,3,3',4,4'-PeCB
2,3,4,4',5-PeCB
2,3',4,4',5-PeCB
2',3,4,4',5-PeCB
2,3,3',4,4',5-HxCB
2,3,3',4,4',5'-HxCB
2,3',4,4',5,5'-HxCB
2,3,3',4,4',5,5'-HpCB
2,2',3,3',4,4',5-HpCB
2,2',3,4,4',5,5'-HpCB

0.0001
0.0003
0.1
0.03
0.00003
0.00003
0.00003
0.00003
0.00003
0.00003
0.00003
0.00003
0.0001
0.00001
        Table 15: Dioxin Toxicity Equivalence Factors.
        * Di-ortho values come from Ahlborg, U.G., et al. (1994), which are the WHO 1994 values from Toxic
        equivalency factors for dioxin-like PCBs: Report on WHO-ECEH and IPCS consultation, December 1993
        Chemosphere. Volume 28. Issue 6. March 1994. Pages 1049-1067.
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Risk Characterization
Using the toxicity factors and the exposure assumptions described previously, the cancer and non-cancer
risks were calculated for each food item consumed at the inland non-anadromous diet rate in each reach.
The risks are summarized in Table 18. The results from each combination of species and reach are
tabulated in Appendix F.  The risk of chemicals with health effects other than cancer, so called non-
carcinogens, is expressed as a Hazard Quotient (HQ), which is the value produced by dividing the
average daily dose at the area of interest by the Reference Dose, which is considered to be the "safe"
dose.  The Reference Dose is derived by EPA, preferably, or other agencies from the scientific literature.
When the average daily dose from the area of interest is less than the Reference Dose, then the HQ will
be less than 1. HQ values of 1 or less are generally considered to be insignificant. A preliminary risk
assessment was conducted as detailed below using the EPA calculator for fish consumption risk-based
concentrations on the EPA Regional Screening Level website (http://epa-prgs.ornl.gov/cgi-
bin/chemicals/csl search .)This computer program calculates the  concentration of a contaminant in food
associated with a given risk level (i.e. a risk-based concentration), using chemicals and  exposure
assumptions that can be entered into the calculator.

Summarized below are the risk-based concentrations (RBCs) for the three ingestion rates and three
chemicals.
Risk Based Concentrations
Ingestion
Rate
(g/day)
70 ( wood duck)
133 (plants)
286 (fish/turtle)
Chemical
TEQ
meHG
PCB
TEQ
meHG
PCB
TEQ
meHG
PCB
Risk Based Concentration (mg/kg)
Cancer
(for Cancer Risk =
1E-06)
1.87E-08
NA,
1.22E-03
9.85E-09
NA
6.40E-04
4.58E-09
NA
2.98E-04
Non-Cancer
(for Hazard
Quotient = 1)
7.30E-07,
1.04E-01,
2.09E-02
3.80E-07
5.49E-02
1.10E-02
1.79E-07
2.55E-02
5.10E-03
Table 16: Risk Based Concentrations.
TEQ = dioxin Toxic Equivalents
meHg = methyl mercury
PCB = Polychlorinated Biphenyls
NA = Not Applicable
For the current preliminary risk assessment, the risk based concentration (RBCs) were calculated for risks
of HQ =1 and CR =1E-06. The contaminants selected from the drop down list on the calculator were 2,
3, 7, 8-TCDD (for total TEQ based on dioxins/furans and dioxin-like PCBs), methyl mercury (for total
and methyl mercury levels), and Aroclor 1254 (as representative of cancer and non-cancer risk of total
PCBs by congener analysis). The biota consumption rates were set at one of the three different tribal
ingestion rates (70 g/day for wood duck, 133 g/day for plants, and 286 g/day for fish, eel, and turtle). The
other exposure assumptions include an exposure frequency of 350 days/yr, an exposure duration of 30
years, adult body weight of 70 kg, lifetime of 70 yr,  non-cancer averaging time of 10950 days (i.e. 30
years x 365 days/year) and cancer averaging time of 25550 days (i.e. 70 years x 365 days/year).
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The equations used in the calculator to calculate RBCs for HQ =1 and cancer risk (CR) = 1E-06 are
provided below:

The equation for calculating the non-carcinogenic screening level is provided below:

   RBC-nc = THQ * AT-nc * BW
           EF * ED * 1/RfDo * IR-F * CF

The equation for calculating the carcinogenic screening level is provided below:

   RBC-c = TR * AT-c * BW
           EF * ED * CSF * IRF * CF

   RBC-nc =Risk Based Concentration, non-carcinogen (mg/kg)-chemical specific
   THQ = Target Hazard Quotient (unitless) (set at HQ=1 for this assessment)
   AT-nc = Averaging Time-non-cancer = 10950 days (365 days/year x 30 years)
   RBC-c = Risk Based Concentration, carcinogen (mg/kg)-chemical specific
   TR =  Target Risk (unitless), set at cancer risk = 1E-06 for this assessment
   AT-c = Averaging Time, cancer =25550 days (365 days/year x 70 years)
   BW = Body Weight = 70 kg
   ED = Exposure Duration = 30 years
   EF = Exposure Frequency =350 days/year
   RfDo = oral Reference Dose (mg/kg-day)-chemical specific
   CSF = Cancer Slope Factor (per mg/kg-day)-chemical specific
   IR-F = Ingestion rate-Fish (mg/day)-food item specific
   CF = Conversion Factor =  1E-06 kg/mg
   *=symbol for multiplication

The risks were calculated differently for cancer and non-cancer risk as described below. For non-cancer
risk, the Hazard Quotient (HQ) was calculated by dividing the concentration of a chemical in biota by the
non-cancer RBC for HQ=1 for that chemical. As an example, if the concentration in biota is 8 mg/kg, and
the RBC representing HQ =1 is 4 mg/kg, then the HQ would be 2 (i.e. 8/4=2).

The Cancer Risk (CR) was calculated by dividing the concentration of a chemical in biota by the cancer
RBC for CR=1E-06 for that chemical,  and then multiplying by  1E-06. As an example, if the
concentration in biota is 4 mg/kg, and the RBC representing CR=1E-06 is 2 mg/kg, then CR would be
2E-06 (i.e. 4/2 x 1E-06 = 2E-06).

Since cancer risks of different chemicals can be added together, the CR of each carcinogenic chemical
was added to calculate a total CR. Some chemicals such as dioxins/furans and PCBs have both cancer
and non-cancer effects.  Non-cancer risks for different chemicals can be added together only if the
chemicals have the same target tissue (e.g. liver damage, central nervous system effects). Therefore, the
HQ of each non-carcinogenic chemical was not added to the HQ of other non-carcinogenic chemicals.

The non-cancer Reference Doses and cancer Slope Factors for the contaminants as used in the calculator
were obtained from the latest version of EPA's Regional Screening Level website, which obtain most of
the toxicity values from the Integrated  Risk Information System (IRIS) available at http://epa.gov/iris/.
Although IRIS does not have a Slope Factor for dioxins/furans, this calculator used EPA's currently
Page | 70                                             Final                     2015

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 recommended value from the California Environmental Protection Agency (CalEPA). Based on the
 conclusion that most of the mercury detected in biota is methyl mercury, the Reference Dose in the
 calculator for methyl mercury was used rather than the Reference Dose for inorganic mercury.
 According to the EPA Integrated Risk Information System (IRIS), the primary toxic effects of mercury
 are non-cancer effects on the neurological system. Mercuric chloride and methyl mercury are classified
 as possible human carcinogens but the cancer risk is not quantifiable due to no or inadequate data in
 humans and limited evidence of carcinogenic effects in animals. As  a result, mercury has a Reference
 Dose (from IRIS) but no Slope Factor for cancer risk. Dioxins/furans have both cancer and non-cancer
 effects and, therefore, the screening level risk assessment used a Reference Dose (from IRIS) and a Slope
 Factor (from CalEPA). PCBs have both cancer and non-cancer effects, and, therefore, the preliminary
 risk assessment used a Reference Dose and Slope Factor for Aroclor 1254, a mixture of PCB congeners.
 Toxicity factors for Aroclor 1254 were selected because it is the only PCB on the IRIS database that has
 both a Reference Dose and a Slope Factor, and it is adequately and conservatively representative of the
 mixture of highly chlorinated PCBs likely to occur in fish populations.

 Risk management Criteria
 The preliminary risk assessment estimates the quantitative level of risk; however, the process of risk
 assessment does not itself identify the maximum acceptable level of risk.  Identifying acceptable risk
 levels is a risk management process that is based on the goals of the  stakeholders as well as any
 regulatory requirements.  Therefore, it is beyond the scope of this report to specify which, if any, state
 or federal regulatory program(s) applies to the issue of contaminants in biota in or near the Penobscot
 River. Nevertheless, the risks identified in this report can be placed in context by identifying the
 maximum permitted risks under some of EPA environmental  regulatory programs as shown in table 17
 below.
EPA Acceptable Cancer Risk Management Criteria
EPA
EPA
EPA
EPA
EPA
EPA
Maximally exposed
member of the
general public
General public
General public
General public
General population
Sensitive
subpopulation
Hazardous air pollution
Drinking water
Abandoned hazardous waste sites
Operating hazardous waste sites
Surface Water Quality Criteria
Surface Water Quality Criteria
lxlO-6to lxlO-4a
Goal of zerob
lxlO-6to IxlO-4
lxlO-6to lxlO-4c
lxlO-6to IxlO-5
lxlO-6to IxlO-4
Table 17: Maximum Lifetime Cancer Risks Permitted by EPA Environmental Regulations

   a.  If the risk to the maximally exposed individual (MEI) is no more than 1 x 10~6, then no further
      action is required. If not, the MEI risk must be reduced to no more than 1 x 10~4, regardless of
      feasibility and cost, while protecting as many individuals as possible in the general population
      against risks exceeding 1 x 10~6.

   b.  EPA sets a goal of zero risk for carcinogens in drinking water. The enforced limit is then set as
      close as possible to this goal given what is feasible using the best available control technology.

   c.  Chemicals are listed as hazardous if they pose a risk of > 1 x 10~5. They are de-listed only if their
      risk is determined to be < 1  x 10~6.  Corrective action must reduce risks to 1 x 10~4 to  IxlO"6.  For
 Page | 71
Final RARE Report August 2015

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         incinerators, risks associated with Group A and Group B carcinogens (substances likely to cause
         cancer in humans or animals) can be no more than 10~6. Risks associated with Group C carcinogens
         cannot exceed 10~5.

This table identifies the maximum lifetime cancer risk under EPA regulations for hazardous air pollution,
surface water quality criteria, drinking water, and operating or abandoned hazardous waste sites. Cancer risk
is expressed as a probability  of getting cancer from the particular type of exposure, for instance a probability
of 1 in 1 million, also expressed mathematically as either 1 x  10~6 or 1E-06 Table 17 indicates that a cancer
risk level of 1 x 10~6 (1 in 1 million) or less is generally considered to be below regulatory concern.

Therefore, for this report, the cancer risks of 1 x 10~6 or less are considered to be of "no concern". The
cancer risk is different from the risk based concentration because the CR is a multiple of the RBC. Cancer
risks of 1 x 10~6 or less, or non-cancer risks of HQ of 1 or less, are designated as being of "no concern".  All
cancer risks  greater than 1 x  10~6, or non-cancer risks of HQ greater than 1, are designated as being of
"potential concern".

This risk management approach using just two criteria of "no concern" or "potential concern" is  similar to
the risk management criteria used by ATSDR and is  considered to be appropriate given the high  uncertainties
associated with the contaminant data in this study. Such uncertainties include the use of only one or two
composite samples per reach for each species, the use of maximum contaminant concentrations rather than
average concentrations, the collection of larger (and therefore probably more contaminated) fish, and the
collection of biota during only one season. Since it is known that contaminants in river fish vary greatly with
species, age, river location/habitat, and season, the actual representative concentrations in biota of the
Penobscot River may be higher or lower than those measured in this study. Additional collection and
contaminant analysis of biota would be necessary to  develop statistically based representative contaminant
concentrations.

The risk of chemicals that have effects other than cancer, such as kidney damage or birth defects, is also
regulated by various environmental regulatory programs.  The non-cancer risk of these chemicals is
expressed as a hazard quotient (HQ), which is simply the number obtained when the estimated exposure dose
of the chemical is divided by the no-effect dose, the so-called Reference Dose (RfD). The Reference Dose is
derived from the scientific literature and published by EPA or other agencies. If the estimated exposure dose
is higher than the no-effect dose, then the HQ will be greater than 1. Therefore, most regulatory  programs
consider a HQ greater than 1 to be of potential concern. For instance, an HQ of 1 is used for EPA drinking
water health advisories and national recommended water quality criteria for protection of human health for
non-carcinogens and as the level above which remedial actions are considered at operating or abandoned
hazardous waste sites.

Based on this analysis of multiple regulatory programs, and without specifying which, if any, regulatory
program applies to contaminants in biota in or near the Penobscot River, risks of 1 x 10~6 or less, or an HQ of
1 or less, are identified in this report to be of "no concern". Conversely, risks of 1 x  10~6 or greater, or an HQ
greater than  1, are identified  in this report to be of "potential concern".
   Page |  72                                                                        201

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Preliminary Risk Results
Using the toxicity factors and the exposure assumptions described previously, the cancer and non-cancer
risks were calculated for each food item consumed at the Inland Non-Anadromous diet rate in each reach.
The risks are summarized in Table 18. The results from each combination of species and reach are tabulated
in Appendix F.

EPA used the data from this study to conduct a screening level risk assessment which compared the
concentrations in biota to risk-based concentrations representing a Hazard Quotient of 1 and a cancer risk
of 1 in 1 million. These risk levels are considered to be insignificant or of "no concern". The results
suggest that that ingestion of each animal species at the Wabanaki Exposure Scenario consumption rates
is associated with a risk higher than the level of "no concern" i.e., HQ=1 and CR=1E-06.

Among the animal biota, the lowest risks were for wood duck, with a maximum HQ of 1 and a maximum CR
of 6E-05. All other animal species had HQ values  greater than 1 and CR values greater than 1E-06. Among
fish, eel, and turtle, the HQ values ranged from a low of 5 for brown  bullhead in the control reach 6 to a high
of 40 in smallmouth bass in four reaches and snapping turtle in two reaches (including control reach 6).
EPA's preliminary risk assessment indicates that the species of highest concern are small mouth bass,
American eel and snapping turtle. These HQ values above one were due primarily to mercury, but also
dioxin TEQ in snapping turtle and eel. The CR values were due primarily to dioxin TEQ and secondarily to
PCBs.  Based on EPA's designation of cancer risks greater than 1E-06 as being of potential concern in this
report, ATSDR's recommendations that PIN members should limit the consumption of eel and snapping
turtles  from the  reaches identified in this study are not inconsistent with EPA's preliminary risk assessment.
PIN members should be aware that reach 5 is an area where there are especially high cancer risks in eel,
snapping turtle,  white perch and brown bullhead (See Figures 9-14).  PIN members can use the information
in Table 18 and Figures 9 to!4 to tailor their fishing, hunting and gathering practices to reduce their health
risks.

The data from this study also showed that consumption of plant materials at the Inland Non-Anadromous
tribal consumption rate had a maximum HQ that was less than 1 and  a maximum CR of 9E-06.  Since the
CR for plant materials is greater than 1E-06, consumption of plants is of "potential concern". EPA's
screening level risk assessment also indicates that mercury was not found in duck, fiddlehead fern,  or
medicinal roots  at levels  of health concern. Table 18 and Figures 9-14 illustrate which species are of
potential  concern for each reach.
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Adult Risks at Inland-Non-Anadromous Tribal Ingestion Rate for Penobscot River Reaches
Tested
Food
Item

Chain
Pickerel
Yellow
Perch
White
Perch
Smallmouth
Bass
Brown
Bullhead
American
Eel
Wood duck
Fiddlehead
Fern
Medicinal
Plant
Snapping
Turtle
Tribal
Ingestion
Rate
(g/day)
286
286
286
286
286
286
70
133
133
286
Risk for Adults
Reach 1
HO
20

20
30
10
30
0.5
<1

40
CR
9E-
06

8E-
05
3E-
05
7E-
05
5E-
04
2E-
05
1E-
06

1E-
03
Driver
TEQ
Hg

Hg
TEQ
TEQ
PCB
Hg
TEQ
Hg
Hg,
TEQ
TEQ


Hg
TEQ
Reach 2
HO
30
20

40
20
30
0.6
<1


CR
1E-
05
8E-
05

4E-
05
3E-
05
3E-
04
3E-
05
2E-
06


)river
Hg
TEQ
Hg
TEQ

TEQ
PCB
Hg
TEQ
Hg
Hg,
TEQ
TEQ
PCB


Reach 3
HO
30
20

40
10
20
0.6
<1
<1
20
CR
9E-
06
1E-
05

3E-
05
6E-
05
5E-
04
3E-
05
4E-
07
6E-
06
2E-
04
Driver
Hg
TEQ
TEQ
Hg

TEQ
PCB
Hg
TEQ
Hg
Hg,
TEQ
TEQ


TEQ
Hg
Reach 4
HO
10
6
20
40
7
10
1
<1
<1
20
CR
8E-
06
7E-
06
9E-
05
6E-
05
7E-
05
3E-
04
6E-
05
5E-
07
9E-
06
5E-
04
Driver
TEQ
Hg
Hg
TEQ
Hg
TEQ
TEQ
PCB
Hg
TEQ
Hg
Hg,
TEQ
TEQ


PCB
TEQ
Hg
Reach 5
HO
20

20
40
20
30


<1
40
CR
3E-
05

2E-
04
4E-
05
2E-
04
1E-
03


2E-
06
6E-
04
Driver
TEQ
Hg

Hg,
TEQ
TEQ
PCB
Hg
Hg,
TEQ
Hg,
TEQ



Hg,
TEQ
Reach 6
HO
20
10
20
30
5
8

<1
<1
10
CR
1E-
05
3E-
06
7E-
05
5E-
05
5E-
05
1E-
04

3E-
07
4E-
06
3E-
05
)river
Hg
TEQ
Hg
TEQ
Hg
TEQ
TEQ
PCB
Hg
TEQ
Hg
Hg,
TEQ



TEQ
Hg
Table 18: Adult Risks at Inland-
TEQ = Dioxin Toxic Equivalents
Hg = Mercury
Non-Anadromous Tribal Ingestion Rate for Penobscot River Reaches
   HQ = Hazard Quotient
   CR= Cancer Risk
Numbers that are bolded and shaded in the column under HQ indicate that there is a non-cancer risk of potential concern because the risk value exceeds HQ=1
and numbers in the column titled CR indicate that there is a cancer risk of potential concern because the risk value exceeds CR = 1E-06.  Numbers that are not
bolded or shaded indicate that the health risk is of no concern because the HQ is 1 or less or the CR is 1E-06 or less.
       Page  74
                                  Final RARE Report August 2015

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                Reach 6
                Reach 5
                Reach 4
                Reach 3
                Reach 2
                Reach 1
                                Risk for Adults by Study Reach
•
                       Fish
American
   Hcl
Wood
Duck
                                                     Plant
                         Snapping
                          Turtle
Figure 9: Chart of Adult Risk by Reach
SBBIdffiBIH  Concentrations in green shading indicate the risk is of no concern.
Yellow Shading  Concentrations in yellow shading indicate there is a non-cancer health risk of concern (HQ>1).
                Concentrations in orange shading indicate there is a cancer health risk of potential concern
(CR>1E-06).
(Note: Half orange and half yellow indicates there is both a non-cancer health risk and a potential cancer risk of
concern.)
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                      Final RARE Report August 2015

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                          Highest contaminant concentrations
                             in sediment (dry weight) and
                           biota (wet weight) by study Reach
Total Dioxin/  Total WHO-    Total        Total
 Furan(17    PCB (12    TEQs (29    (142) PCB
 Congeners)   Congeners)   Congeners)    Congeners)
                                                                 Methyl
                                                                 Mercury
                                                                          Mercury
              Reach 5
             Reach 4
             Reach 3
             Reach 2
             Reach 1
                      1 isli
      American   Wood
        i;el     Duck
                                                 Plan,
                                                                  Sediment
Figure 10: Chart of highest contamination concentrations in sediment and biota by Reach.
             | Concentrations in green shading indicate there is no health concern.
Yellow Shading Concentrations in yellow shading indicate there is a non-cancer health risk of concern (HQ>1).
             | Concentrations in orange shading indicate there is a cancer health risk of concern (CR>1E-06).
(Note: Half orange and half yellow indicates there is both a non-cancer and a cancer risk of concern. BB=Brown
Bullhead;WP=White Perch)
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                                  Final RARE Report August 2015

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Hazard Quotients
Figures 11 to 12 illustrate the Hazard Quotient for each animal species that exceed a HQ =1 in each
reach. A hazard quotient of 1 or less is considered to represent a non-cancer risk of "no concern". The
hazard quotient is the value of the estimated dose of contaminants in flora and fauna divided by the safe
ingestion dose for adverse health effects other than cancer.
             Hazard Quotients of  Potential  Concern
            Reach 1
Reach 2
Reach 3
Reach 4
               * Chain Pickerel   • Yellow Perch
                  i White Perch
Reach 5
                   - Smallmouth Bass
Control Reach
               s Brown Bullhead  "American Eel    • Snapping Turtle
Figure 11: Hazard Quotient (HQ) for fauna that exceed a HQ =1.
Reach 1 = Milford Dam Impoundment (MIL)        Reach 4 = Mohawk-Mattaseunk Dam (MM)
Reach 2 = Sunkhaze-West Enfield Dam (SWE)      Reach 5 = Mattaseunk Dam Impoundment (MAT)
Reach 3 = West Enfield Dam Impoundment (WEI)   Reach 6 = Control Reach, East Branch-Salmon Stream Lake (EBS)
                         HQ of Potential Concern
                          per Species per Reach
                  Reachl   - Reach 2  • Reach 3  * Reach 4  » Reach 5  - Control Reach
 Figure 12: Hazard Quotient (HQ) for fauna that exceed a HQ =1.
Page  77
                         Final RARE Report August 2015

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                       AE- American Eel
                       CP- Chain pickerel
YP- Yellow perch
WP- White perch
BB- Brown bullhead
 8MB- Smallmouth bass
 ST- Snapping turtle
Cancer Risk
Figure 13 illustrates the Cancer Risk (CR) for fish, eel, and snapping turtle that exceed a CR= 1E-06 for
each reach. The CR is expressed as a lifetime probability of getting cancer due to exposure at the area
of interest, over and above the normal probability of getting cancer from all causes.
 Figure 13: Cancer Risk (CR) for fauna that exceed a CR= 1E-06.
 ST- Snapping turtle   AE- American Eel
 WP-White perch     BB- Brown bullhead
                      Plant Material
              CR of Potential  Concern
      1.0E-05
      9.0E-06
      8.0E-06
      7.0E-06
      6.0E-06
      5.0E-06
      4.0E-06
      3.0E-06
      2.0E-06
      1.0E-06
              Reach 1   Reach 2   Reach 3   Reach 4  Reach 5   Reach 6
                                Medicinal plant
Figure 14: Cancer Risk (CR) for flora that exceed a CR= 1E-06.
Reach 1 = Milford Dam Impoundment (MIL)          Reach 4 = Mohawk-Mattaseunk Dam (MM)
Reach 2 = Sunkhaze-West Enfield Dam (SWE)        Reach 5 = Mattaseunk Dam Impoundment (MAT)
Reach 3 = West Enfield Dam Impoundment (WEI)     Reach 6 = Control Reach, East Branch-Salmon Stream Lake
                                                      (BBS)
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          Final RARE Report August 2015

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Uncertainty Analysis
There are many sources of variability and uncertainty in conducting any risk assessment.  The
variability in concentrations among individual fish is unknown because only composites were analyzed.
There is uncertainty about the concentrations of the individual dioxin/furan/PCB congeners because of
analytical detection limits.  The analyses were conducted using the best available practicable methods;
therefore the total concentrations of the congeners and TEQs may be somewhat under- or over-
estimated, but the data are usable for a preliminary risk assessment. The use of maximum
concentrations for preliminary risk assessment suggests that the risks are somewhat over-estimated.
Since the maximum concentration of the composite sample was used, there were individual fish in the
sample that had lower and higher concentrations.  It is likely that some individual fish in the river have
even higher concentrations than the maximum composite concentration, particularly the larger and
older fish that may be caught in the river.

There is uncertainty about the exposure assumptions in that some people may eat more or less than the
amount used in the preliminary risk calculation. It is assumed that an adult tribal member weighs 70 kg
and would ingest the individual food items for 30 years in the study area over a lifetime of 70 years. It
is probable that some tribal members would not move from the area and therefore would have 70 years
of exposure over a lifetime of 70 years. Use of the latter assumption would approximately double the
risks (i.e. 70/30= 2.3). It should also be noted that this preliminary risk assessment was for adults
exposed from childhood through 30 years of age, rather than for children exposed only during
childhood.  Tribal consumption rates for children were not available. Assuming that a 6-year old child
(weighing 15 kg) would consume about half of the adult consumption rate for 6 years, the non-cancer
risks for a child would be about twice as high as those for a 70 kg adult consuming at the adult
consumption rate for 30 years.

It should be emphasized that the preliminary risk assessment is based on the specified tribal
consumption rate of individual food items,  rather than the combined consumption of different kinds of
food items. Since there is an almost infinite number of combinations possible for consumption of the
seven animal  species and plant species, estimation of combined risks was beyond the scope of this
preliminary risk assessment. Rather, the risks of each food item at the tribal consumption rate were
estimated in this study to help enable individual tribal members to evaluate the  risks for the particular
combinations of food items that they consume.

There is also uncertainty about whether the non-cancer risks of dioxins/furans,  PCBs, and mercury
should be added together.  All three chemicals have neurological effects but there are many other toxic
effects that these chemicals do not have in  common. It would be conservative to add the hazard
quotients together but this was not done; however, inspection of the HQ values in the risk table in
Appendix F indicates that adding the HQ values would not change the conclusion about whether the
separate HQ values exceed 1 or not.
Page  79                                             Final RARE Report August 2015

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A
         Health and Exposure Assessment Conclusions and
                               Recommendations

TSDR Health Assessment
ATSDR used the data generated from the RARE report to conduct a health assessment for the
Penobscot Indian Nation. A Health Assessment is a way for ATSDR to respond to a need for health
information on toxic substances and to make recommendations for actions to protect the public's
health.

ATSDR staff evaluated information available about toxic material at the site, determined whether
people might be exposed to it, and reported what harm exposure might cause.

Health Assessments typically consider the following:
> what the levels (or "concentrations") of hazardous substances are;
> whether people might be exposed to contamination and how (through "exposure pathways" such
   as breathing air, drinking or contacting water, contacting or eating soil, or eating food);
> what harm the substances might cause to people (or the contaminants' "toxicity");
> whether working or living nearby might affect people's health; and,
> other dangers to people, such as unsafe buildings, abandoned mine shafts, or other physical
   hazards.

Based on the results of the samples collected, the ATSDR came to the following conclusions
concerning the health hazards:
> Penobscot Indian Nation (PIN) members who eat fish and turtle at the ingestion levels suggested
   in the Wabanaki Traditional Cultural Lifeways Exposure Scenario report (Scenario) may be
   exposed to harmful levels of mercury, dioxins/furans and dioxin-like PCBs.

> ATSDR is most concerned about mercury in fish and turtle taken from the Penobscot River.
   Mercury is most harmful to children and developing fetuses, therefore  it is especially important for
   pregnant and breastfeeding women, women who may become pregnant, and children to limit their
   consumption offish and turtle in order to decrease their risk of neurological damage due to mercury
   exposure.

> PIN members who eat duck, fiddlehead fern, or medicinal plants at the Scenario- suggested
   ingestion rates will not be exposed to harmful levels of mercury, PCBs, dioxins/furans or dioxin-
   like PCBs.

> Incidental ingestion of, and dermal exposure to, Penobscot River sediment does not pose a
   human health hazard. All sediment contaminants analyzed in this report were found in
   concentrations below initial screening values with the exception of dioxins/furans in three
   samples. Dioxin/furan concentrations in those three sediments were below human health
   exposure guidelines and therefore pose no health threat to the Penobscot Indian Nation tribal
   members that may be exposed to sediments in the Penobscot River.
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ATSDR 's Health Assessment Recommendations
ATSDR is most concerned about mercury in fish and snapping turtle taken from the Penobscot
River. Mercury is most harmful to children and developing fetuses, therefore it is especially
important for pregnant and breastfeeding women, women who may become pregnant, and children
to limit their consumption offish and snapping turtle in order to decrease their risk of neurological
damage due to mercury exposure. PIN members should follow the existing PIN DNR fish advisory
and the State of Maine Safe Eating Guidelines for all fish caught in the Penobscot River.

ATSDR recommends that Penobscot Indian Nation members should reduce their consumption of
fish and snapping turtle in order to decrease their exposure to potentially harmful methyl mercury,
as well as dioxins/furans and dioxin-like PCBs based on the following recommendations:
> PIN members limit their consumption offish to 1-2 fish meals per month in order to minimize
   their risk of harmful health effects due to methyl mercury; and their lifetime risk of cancer due
   to dioxin/furans and dioxin -like PCBs.

> PIN members limit their snapping turtle consumption to 2-3  servings per month.

> If PIN members eat both fish  and turtle, limit their consumption to no more than some
   combination of 1-2 (10 oz.) servings offish, OR 2-3 (8 oz.) servings of turtle per month.

> Incidental ingestion of, and dermal exposure to, sediment in the Penobscot River is not expected
   to cause a health hazard.

> It is safe to eat wood duck, fiddlehead ferns and medicinal roots at the rates suggested in the
   Wabanaki Traditional Cultural Lifeways Exposure Scenario.

US EPA Exposure Assessment Recommendations
The data from this study were used by EPA in a preliminary risk assessment which compared the
concentrations in biota to risk-based concentrations representing a Hazard Quotient of 1 and a cancer
risk of 1 in  1 million. The risks are summarized in Table 18 and Figures 9 to 14.

The results  indicate that consumption of plant materials at the Inland Non-Anadromous tribal
consumption rate had a maximum HQ that was less than 1 and a maximum CR of 9E-06.  Among the
animal biota, the lowest risks were for wood duck, with a maximum HQ of 1 and a maximum CR of
6E-05. All  other animal species had HQ values  greater than 1. Among fish, eel, and turtle, the HQ
values ranged from a low of 5 for brown bullhead in the control  reach 6 to  a high of 40 in smallmouth
bass in four reaches and snapping turtle in two reaches (Reaches 1 and 5).  These HQ values above one
were due primarily to mercury, but also dioxin TEQ in snapping turtle and eel.

All the fish, eel and turtle analyzed for this study exceeded the CR of Ix 10"6  and have a cancer risk of
potential concern.  The CR values for these animal species were due primarily to dioxin TEQ and
secondarily to PCBs. Based on EPA's preliminary risk assessment, the species of highest concern are
Smallmouth Bass,  White Perch, Brown Bullhead, American Eel and  Snapping Turtle. Table 18 and
Figures 9 to 14 illustrate which species are of most concern for HQ and CR per reach.

Based on EPA's assessment of cancer risks, EPA concurs with ATSDR that PIN members should
limit the consumption of eel and snapping turtles from the reaches identified in this study and that
the consumption of plant material sampled in this study may pose a risk of potential concern.

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However, the risk from consuming the plant material is less than ten times the level considered to
be of "no concern".  Since EPA's screening level risk assessment is based on a maximum
acceptable risks as defined by various EPA regulatory programs and not based on health based
standards as is ATSDR's Public Health Assessment, some of the conclusions between ATSDR and
EPA differ. For example, EPA's screening level risk assessment did show that there is a cancer risk
of "potential concern" in wood duck and plants at certain reaches in the Penobscot River,
particularly reach 4, which do not mirror the conclusions of ATSDR's. This is due to the different
methodologies used by the different agencies and the designation of risk management criteria of
"no concern" defined as HQ=1  and CR= 1E-06. Nonetheless, EPA does concur with ATSDR's
recommendations that the consumption of plants and wood duck do not pose a significant health
risk while the consumption offish, especially eel, and snapping turtle should be limited per
ATSDR's recommendations  PIN members should be aware that reach 5 is an area where there are
especially high screening cancer risks in eel, snapping turtle, white perch and brown bullhead (See
Figure 13). If PIN members use the information in Table 18 and Figures 9-14 to tailor their fishing,
hunting and gathering practices this will help to reduce the risk of cancer and non-cancer health
effects for PIN members.
Recommendation for further Investigation
Because this study was a preliminary assessment, it was understood from the beginning that only a
limited number of samples could be collected and analyzed. Samples of edible muscle and plant issue
were analyzed for a screening level human health risk assessment.  The remaining tissue (offal) was
frozen so that the "whole body" contaminant concentrations could be mathematically reconstructed for
evaluation of ecological risk through food chain transfer (e.g. fish to fish-eating bird).  The data from
the preliminary study supports a conclusion that contamination levels are high enough in specific fauna
in certain reaches to warrant further investigation of both human health and ecological risk.

Due to the culturally significant use of, and subsistence on, these resources, and the potential for
adverse ecological effects due to food chain bioaccumulation, EPA recommends that a more thorough
research study be conducted.  Such a study should include collecting and analyzing sufficient individual
fish to statistically characterize how contaminant concentrations are related to species, individual
length/weight, and with river location.  These relationships can be used to provide risk-based
recommendations to PIN members concerning consumption offish from different river locations. The
frozen "offal" samples should also be analyzed to estimate the contaminant concentrations in the whole
fish that are consumed by fish-eating wildlife (e.g. eagles, mink, snapping turtles) so that the ecological
risk to such higher trophic level predators can be evaluated.

The resulting data could be used to inform food chain ecological risk assessment on the river, as well
as risk management concerning risk-based size limits, advisories concerning fishing in specific river
locations, and serve as a baseline for tracking changes in contaminant concentrations over
time.  Further studies should be coordinated with the PIN Health Department in their effort to correlate
the health results with fish consumption and track changes in fish consumption behavior through
education and issuance of health advisories to PIN members.
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Conclusions and Recommendations of Mutagenicity Study
Analytical Results of Salmonella Mutagenicity Study
As discussed in the Introduction, the Salmonella mutagenicity assay is the bioassay of choice for
determining the mutagenicity of organic compounds present in environmental media (Claxton et al.
2010). A positive result suggests the possibility that the water or sediment may contain some potential
carcinogens. A negative result has less meaning than a positive result but would suggest that the
mixture or compound has a lower probability of containing carcinogens than would those that produce
a positive response.

A positive mutagenic response is defined as one in which the extract produces a dose-related
increase of at least twofold over the DMSO control number of revertants/plate. Revertants are
colonies of bacteria on the Petri plate; they are mutant bacteria formed from exposure of the
bacterial cells to the extract.  The extracts are tested in the presence and in the absence of a
homogenate of rat liver called S9, which provides some aspects of mammalian metabolism.
Bacteria do not have as much of the enzymatic activities as those found in humans, so S9 provides
some of this activity.  Some environmental mutagens/carcinogens require metabolism in order to be
mutagenic/carcinogenic; thus, S9 is added to the Petri dish to provide this activity. When extracts
are mutagenic in the absence of S9, this indicates that the mutagens in the extract do not require
metabolism and are directly acting on the DNA in the bacteria. Various strains of bacteria are used
in the mutagenicity assay because each strain detects only a limited set of chemical classes of
mutagens. Because no single strain detects all classes of mutagens, a variety of strains are used to
capture mutagenic activity over a range of classes of chemical mutagens that might be in the
extracts.

The drinking water samples from all three sampling days were mutagenic in TA98 -S9, with an
average mutagenic potency in TA98 -S9 of 198 rev/L-eq (Table 19).  Samples from day 8/03 were
positive in TA100 -S9. The average mutagenic potency for TA100 -S9 was 476 rev/L-eq. Blank
XAD samples were not mutagenic (data not shown).

Mutagenic activity was not detected in the majority of the river water samples tested in YG1041
and YG1042 with or without  S9 (Table 20).  The sample "At," which was derived by  pooling 5,
2.37-L samples taken at the outfall from the Lincoln Paper and Tissue Mill, was mutagenic in both
YG1041 +/-S9 and YG1042 -S9 only on the third day of sampling. The resulting average
mutagenic potencies for YG1041 were 144 rev/L-eq -S9 and 210 rev/L-eq +S9. The same "At"
sample was mutagenic in strain YG1042.  The other two sampling days from the outfall ("At") and
the other sampling sites (labeled "Above" and "Below") were negative in both strains and S9
conditions.  Blank XAD samples were not mutagenic (data not shown).

Our results show that the Penobscot River water samples have no or low mutagenic activity for the
classes of compounds that this assay detects relative to that of other river waters (Ohe et al., 2004).
We compared our river water results to the rankings identified in a compilation of surface water
quality monitoring (Umbuzeiro et al., 2001), a review of surface water mutagenicity studies (Ohe et
al., 2004), and the mutagenic-potency classification of industrial wastes and effluents  by Houk
(1992).  The average mutagenic potency of the PIN river water samples (177 rev/L-eq) was <500
rev/L-eq, categorizing the Penobscot River as has having low mutagenic potency. For comparison,
high would be >5,000 rev/L-eq (Ohe et al. 2004).


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The drinking-water samples from all 3 sampling days were mutagenic in TA98 -S9, with an average
mutagenic potency in TA98 -S9 of 198 rev/L-eq (Table 19).  Samples from day 8/03 were positive in
TA100 -S9. The average mutagenic potency for TA100 -S9 was 476 rev/L-eq. Blank XAD samples
were not mutagenic (data not shown).

The drinking water samples exhibited negative or low mutagenic potencies for the classes of
compounds that this assay detects relative to other drinking water samples (DeMarini et al., 1995;
Schenck et al., 1998; Takanashi et al., 2009).  The average mutagenic potency for the positive drinking
water samples in this study was 337 rev/L-eq; most samples had negative results.  For comparison,
Takanashi et al. (2009) found an average mutagenicity of  1,100 rev/L-eq among 179 water samples
from 17 sampling  sites located from Hokkaido to Kagoshima Prefecture, Japan. Compared to the
potencies reported in other studies (DeMarini et al., 1995; Schenck et al., 1998; Takanashi et al., 2009),
the average mutagenic potency of the drinking water samples (337 rev/L-eq) was lower than typical
drinking waters described in the papers above, which are -1000 rev/L-eq.

Results from sediment samples tested in TA98, TA100, YG1041, and YG1042 with and without S9
were mostly negative (Table 21). Positive results for this  group of samples were found in the
"Above" location in YG1041 +S9 and YG1042 -S9, which gave values of 276 and 333 rev/g-eq,
respectively.  The  "Island"  sample was mutagenic in YG1041 -S9 (150  rev/g-eq) and YG1042 -S9
(314 rev/g-eq).  Mutagenic potencies for the sediment samples in all strains ranged from 96 to 333
rev/g-eq (Table 21).

As stated above, the mutagenic potencies for the sediment samples tested were also negative or low
relative to other sediments (Chen and White, 2004) for the classes of compounds that this assay detects.
As noted in Table  21, the positive samples were from the "Island" and two from the "Above" location.
The average mutagenic potencies of the sediment extracts (244 rev/g-eq) were typical of sediments
from urban/industrial areas, which average -150 rev/g-eq  (Chen and White, 2004). For comparison,
sediments from remote regions or heavily contaminated regions have potency values of 10 or >10,000
rev/g-eq, respectively (Chen and White, 2004).  Thus, the  river sediment from the PIN was generally
not mutagenic, but when positive, samples had mutagenic potencies typical of that from
urban/industrial areas as described in the literature.

A second set of samples (data not shown) were captured when river water levels were lower than
the initial sampling period in order to see if the river volume was affecting the results. The sample
set consisted of surface water from the "Below" and "At"  locations and were tested with YG1041
and YG1042 with  and without S9; all were negative.
Summary of Mutagenicity Drinking Water Samples (rev/L-eq)

Strain
TA98
TA100



Exp. date
10/23/09
10/14/09
10/23/09
12/01/09
Sampling date
7/30/09
217
Na
ISb
ISb
8/03/09
195
425
793
211
8/05/09
182
Na
Na
ISb
Table 19: Summary of mutagenicity (rev/L-eq) -S9 of 3 samples of drinking water.
aN = Negative (not mutagenic).
bIS - insufficient sample to test.
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Summary of Mutagenicity of Surface Water Samples (rev/L-eq)

Strain
YG1041



YG1042


Exp.
date
08/25/09
08/25/09
09/01/09
09/01/09
09/10/09
09/10/09

S9a

-
+
-
+
-
+
Sampling Date and Site
Above
7/30/09
Nb
Nb
Nb
Nb
Nb
Nb
8/03/09
Nb
Nb
Nb
Nb
Nb
Nb
8/05/09
Nb
Nb
Nb
Nb
Nb
Nb
At
7/30/09
Nb
Nb
Nb
Nb
Nb
Nb
8/03/09
Nb
Nb
Nb
Nb
Nb
Nb
8/05/09
180
227
108
192
179
Nb
Below
7/30/09
Nb
Nb
Nb
Nb
Nb
Nx
b
8/03/09
Nb
Nb
Nb
Nb
Nb
Nb
8/05/09
Nb
Nb
Nb
Nb
Nb
Nb
Table 20: Summary of mutagenicity (rev/L-eq) of 3 samples of surface water from each of 3 sites.
aS9 = A homogenate of rat liver added to provide mammalian metabolism to the bacteria.
aN = Negative (not mutagenic). Values are given only for positive mutagenic results.
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Summary of Mutagenicity of River Sediment Samples (rev/g-eq)
Strain
TA98

TA100

YG1041


YG1042


Exp date
10/23/09
10/14/09
10/23/09
10/23/09
10/14/09
10/23/09
1 1/06/09
1 1/06/09
11/19/09
1 1/06/09
1 1/06/09
11/19/09
S9a
-
+
-
+
-
+
-
-
+
-
Site
Above
Nb
Nb
Nb
Nb
Nb
276
Nb
Nb
Nb
333
At
Nb
Nb
Nb
Nb
Nb
Nb
Nb
Nb
Nb
Nb
Below
Nb
Nb
Nb
Nb
Nb
Nb
Nb
Nb
Nb
Nb
Island
Nb
Nb
Nb
Nb
96
Nb
203
Nb
Nb
314
Table 21: Summary of mutagenicity (rev/g-eq) of composite river sediments from 4 sites.
aS9 = A homogenate of rat liver added to provide mammalian metabolism to the bacterial cells.
bN = Negative (not mutagenic).
 Conclusions of Salmonella Mutagenicity Study
 The Penobscot River is a valuable resource to the Penobscot Indian Nation and has played a major role in
 their cultural traditions of hunting and fishing. Any threat of contamination to the river will be a concern
 for tribal members.  There have been improvements to the water quality as shown in an Agency for Toxic
 Substances and Disease Registry's (ATSDR) review (Williams and Cseh, 2007) of tissue samples from fish
 caught in the Penobscot River near the town of Lincoln, Maine (upstream from Indian Island) spanning
 1988 to 2003.  These samples showed a slight decrease in the toxic equivalency quotient concentrations of
 dioxins/furans, but a slight increase in the levels of methyl mercury. This may be due in part to some
 changes in the processes of the Pulp and Paper Mills (U.S. EPA, 2007). However, there are fish advisories
 in place for the Penobscot River near Lincoln for dioxins and PCBs and throughout the river for mercury
 regarding fish consumption limits based on findings by the Maine Bureau of Health and the PIN.  The
 ATSDR review was in agreement with these  advisories.

 Our findings in this survey study do not address these issues because of the limitations of the assay, but
 they do show that the surface water, sediment, and drinking water samples evaluated here are either not
 mutagenic or have low mutagenic potencies.  The results indicate that there is little risk to human health
 due to the presence of typical organic mutagens or genotoxic carcinogens, such as PAHs, aromatic amines,
 heterocyclic amines, or nitroarenes, which are readily detectable by the Salmonella mutagenicity assay.

 Determining the actual source(s) and compound(s) responsible for the low levels of mutagenicity detected
 would require a more rigorous and much larger study than the present one. Surface water is a complex
 mixture, and assessing the risk is a complicated puzzle to solve.  Donnelly et al. (2004) discussed the
 challenges in estimating potential health effects associated with complex mixture exposures and concluded
 that extensive information is needed regarding mixture  interactions and the effects of unidentified
 chemicals in the mixture in order to properly  assess the risks.
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Salmonella Mutagenicity Recommendations
The results indicate that there is little risk to human health due to the presence of typical organic
mutagens or genotoxic carcinogens, such as PAHs, aromatic amines, heterocyclic amines, or
nitroarenes, which are readily detectable by the Salmonella mutagenicity assay.

However, the assay does not detect non-genotoxic carcinogens or certain other types of toxicants such
as PCBs, dioxins, most metals, neurotoxins, or developmental toxins, which might be present in the
water or sediment.  In addition, our study did not evaluate any airborne toxicants or toxicant exposure
associated with lifestyle exposures.  Given the limits of the assay and of our study, there appears to be
either no or low levels of mutagenic activity in the river and drinking water due to typical genotoxic,
organic compounds.  Other types of assays and analyses are required to identify the presence of
dioxins, PCBs, and metals that might contaminate the water or air.
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                                  APPENDIX A
                             ABBREVIATIONS
ATSDR
BEAD

BIA
CalEPA
CAFRL
CERC
CD
COPC
CR
CV
CWA
DI
Dioxin
DMSO
DNA
DOT
DQO
ECL
EPA
EPANE
ESAT
FGS
Furan
FTP
Hg
HQ
HRGC/HRMS
IRIS
IUPAC
LP&P
LSC
MEDEP
NCEA
NERL
NFRAP
NWS
OP
OPP
ORD
PA/SI
PAHs
PCB
Agency for Toxic Substances and Disease Registry
Biological and Economic Analysis Division, Environmental Chemistry,  Office
of Pesticide Programs, US EPA
Bureau of Indian Affairs
California Environmental Protection Agency
S.O. Conte Anadromous Fish Research Laboratory
Columbia Environmental Research Center
Compact Disk
Chemical of Potential Concern
Cancer Risk
Health-based comparison value
Clean Water Act
Deionized
Polychlorinated dibenzo(p)dioxin
Dimethylsulfoxide
Deoxyribonucleic acid
Department of the Interior
Data Quality Objective
Environmental Chemistry Laboratory
U.S. Environmental Protection Agency
Environmental Protection Agency New England
Environmental Services Assistance Team
Frontier Geosciences Inc.
Polychlorinated dibenzofuran
File Transfer Protocol
Mercury
Hazard Quotient
High Resolution Gas Chromatography/High Resolution Mass Spectrometry
Integrated Risk Information System
International Union of Pure and Applied Chemistry
Lincoln Pulp and Paper
Leetown Science Center
Maine Department of Environmental Protection
National Center for Environmental Assessment
National Environmental Research Laboratory, US EPA
No Further Federal Remedial Action Planned
National Weather Service
Operating Procedures
Office of Pesticide Programs
Office of Research and Development
Preliminary Assessment/Site Investigation
Polycyclic aromatic hydrocarbons
Polychlorinated biphenyl
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PCDD          Polychlorinated dibenzo(p)dioxin (dioxin)
PCDF           Polychlorinated dibenzofuran (furan)
PFK           Perfluorokerosene
PIN             Penobscot Indian Nation
PIN-DNR       Penobscot Indian Nation-Department of Natural Resources
POTW          Publically owned treatment works
PQAM          Program QA Manager
PQL            Project Quantification Limits
PTFE          Polytetrafluoroethylene
QAPP           Quality Assurance Project Plan
QA             Quality Assurance
QC             Quality Control
RARE          Regional Applied Research Effort
SIM           Selective ion monitoring
SOP            Standard Operating Procedure
START         Superfund Technical Assessment and Response Team
SVOC          Semi-volatile organic compounds
SWAT          Surface Water Ambient Toxics Monitoring Program
TEF            Toxic Equivalency Factor
TEQ            Dioxin Toxic Equivalent
TQL            Targeted Laboratory Quantification
TSA            Technical System Audit
TOC            Total Organic Carbon
UMAECL       University of Maine Environmental Chemistry Laboratory
US EPA         United States Environmental Protection Agency
USF&WS       United States Fish and Wildlife Service
USGS           United States Geological Survey
OPP            Office of Pesticide Programs,
VOC           Volatile organic compounds
WHO           World Health Organization
WQS           Water Quality Standards

                                Measurement Abbreviations
°C              degrees Centigrade
°F              degrees Fahrenheit
ft3              cubic feet
km             kilometers
km2             square kilometers
MW            megawatt
mi              miles
mi2             square miles
Hg/g            micrograms per gram (parts-per-million)
mg/kg           milligrams per kilogram (parts-per-million)
mm             millimeters
ng/g            nanograms per gram (parts-per-billion)
ng/kg           nanograms per kilogram (parts-per-trillion)
pg/g            picograms per gram (parts-per-trillion)
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                                         APPENDIX B
                                        REFERENCES

1.    Advances in Modern Environmental Toxicology, Volume XXI, Princeton Scientific Publishing Co.,
     Princeton, NJ 1992
2.    Analysis of Cancer Registry Data, Final Report, Penobscot Indian Nation, 1059-1993. Prepared by Daniel
     S. Miller, Epidemiology and Statistics Branch, Division of Cancer Prevention and Control, Centers for
     Disease Control and Prevention.  May 1994.
3.    ATSDR Public Health Assessment, See ATSDR's website http://www.atsdr.cdc.gov/.
4.    Claxton, L.D., 1985. Assessment of bacterial bioassay methods for volatile and semi-volatile compounds
     and mixtures. Environ Intl 11, 375-382.
5.    Claxton, L.D., 1997. The development, validation, and analysis of Salmonella mutagenicity test methods
     for environmental situations, in: Wells, P.G., Lee, K., and Blaise, C. (Eds.), Microscale Testing in Aquatic
     Toxicology, CRC Lewis Publishers, Boca Raton, Florida, pp. 591-605.
6.    Claxton L.D., Umbuzeiro G.D., DeMarini D.M., 2010. The Salmonella mutagenicity assay: the
     stethoscope of genetic toxicology for the 21st century.  Environ. Health Perspect. 118, 1515-1522,2010.
7.    Claxton, L.D. and DeMarini, D. EPA Intramural Research Protocol for NHEERL/ECD Study: Use of in
     Vitro Genotoxicity Tests to Characterize Cancer-related Toxicants for Water and Sediment-borne
     Environmental -Contaminants in the Penobscot Watershed Region, June 10, 2009.
8.    deFur, P.L., Evans, G.W., Cohen Hubal, E.A., Kyle, A.D., Morello-Frosch, R.A., Williams, D.R., 2007.
     Vulnerability as a Function of Individual and Group Resources in Cumulative Risk Assessment. Environ.
     Health Perspect.  115, 817-824.
9.    desJardins Anderson, T., MacRae, J.D., 2006. Polybrominated diphenyl ethers in fish and wastewater
     samples from an  area of the Penobscot River in Central Maine. Chemosphere 62, 1153-1160.
10.  Di-ortho values come from Ahlborg, U.G., et al. (1994), which are the WHO 1994 values from Toxic
     equivalency factors for dioxin-like PCBs: Report on WHO-ECEH and IPCS consultation, December 1993
     Chemosphere. Volume 28. Issue 6. March 1994. Pages 1049-1067.
11.  Ferrario, J., Byrne, C., Dupuy, A.E., 1997, Background Contamination by Coplanar Polychlorinated
     Biphenyls (PCBs) in Trace Level High Resolution Gas Chromatography (HRGC)/High Resolution Mass
     Spectrometry (HRMS) Analytical Procedures, Chemosphere 34 (11):2451-2466.
12.  Harper B. and Ranco D., 2009. Wabanaki Traditional Cultural Lifeways Exposure Scenario. Prepared
     for EPA in collaboration with the Maine Tribes by Drs. Barbara Harper and Darren Ranco. Available:
     http://www.epa.gov/regionl/govt/tribes/pdfs/DITCA.pdf [accessed 9 February 2011].
13.  Kamps, L. R., Carr, R., and Miller, H. 1972. Total mercury-monomethylmercury content of several species
     offish. Bulletin of Environmental Contamination and Toxicology 8(5): 273-279.
14.  Lazorchak, J.M., McCormick, F.H., Henry, T.R., Herlihy, A.T., 2003.  Contamination of Fish in Stream of
     the Mid-Atlantic Region: An Approach to Regional Indicator Selection and Wildlife Assessment.
     Environ. Toxicol. Chem. 22 (3), 545-553.
15.   Miller DS.  1994. Final Report, Analysis of Cancer Registry Data, Penobscot Indian Nation,  1959-1993, May
     2004. Atlanta: Centers for Disease Control and Prevention.
16.  Miller DS and Drabant B.  1996. Letter clarifying Analysis of Cancer Registry Data for the Penobscot Indian
     Nation. October 8, 1996. Atlanta: U.S. Department of Health and Human Services.
17.  Ohe, T., Watanabe, T., Wakabayashi, K., 2004.  Mutagens in surface waters: a review. Mutat. Res. 567,
     109-149.

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18.   Penobscot Nation Health Department Cancer Registry Report, 1980-1994.  Prepared by Hope Valcarcel,
     Health Records Manager.
19.   Penobscot Nation Health Department Cancer Registry Report, 1994-1998.  Prepared by Hope Valcarcel,
     Health Records Manager.
20.   Fob/chlorinated Dibenzo-/>ara-Dioxins and Polychlorinated Dibenzofurans, Vol. 69, Aug. 1997.  World
     Health Organization, International Agency for Research on Cancer (IARC) Monographs on the Evaluation
     of Carcinogenic Risks to Humans.
21.   Public Health Consultation. Penobscot River, Maine. June 1, 2006. US Department of Health and Human
     Services, Agency for Toxic Substances and Disease Registry, Division of Toxicology and Environmental
     Medicine, Emergency Response Team, Atlanta, GA 30333.
22.   Smith, Andrew E. Evaluation of the Health Implications of Levels of Polychlorinated Dibenzo-p-Dioxins
     (Dioxins) and Polychlorinated Dibenzofurans (Furans) in Fish from Maine Rivers.  Jan. 2008.
     Environmental and Occupational Health Programs, Maine Center for Disease Control, Maine Department
     of Health and Human Services.
23.   Summary Report for the ATSDR Expert Panel Meeting on Tribal Exposures to Environmental
     Contaminants in Plants. Mar. 23, 2001. Prepared for: The Agency for Toxic Substances and Disease
     Registry, Division of Health Assessment and Consultation, Office of Tribal Affairs, Atlanta, Georgia
     (Contract No. 205-95-0901, Task Order No. 29). Prepared by: Eastern Research Group, 110 Hartwell
     Avenue, Lexington, MA 02421-3136.
24.   Survey of the mutagenicity of surface water, sediments, and drinking water from the Penobscot Indian
     Nation, Chemosphere 120 (2015) 690-696, Chemosphere journal homepage
     www.elsevier.com/locate/chemosphere
25.   (LP&P, 2003) - US EPA 2003, Final Preliminary Assessment/Site Inspection Report For Lincoln Pulp &
     Paper Co. Lincoln, Maine, May 30, 2003 (LP&P, 2003).
26.   USEPA, 2004, Exposure and Human Health Reassessment of 2,3,7,8-Tetrachlorodibenzo-p-dioxin
     (TCDD) and Related Compounds. National Academy Sciences (External Review),  United States
     Environmental Protection Agency, Washington, D.C., EPA/600/D-OO/OOCb.
27.   Valcarcel H.  1994. Penobscot Nation Health Department Cancer Registry Report  1980-1994.
28.   Van den Berg, M., Birnbaum, L.S.,  Denison, M., DeVito, M., Farland, W.,  Feeley,  M. Fiedler, H.,
     Hakansson, H., Hanberg, A., Haws, L., Rose, M., Safe, S., Schrenk, D., Tohyama, C., Tritscher, A.,
     Tuomisto, J., Tysklind, M., Walker, N., Peterson, RE.  2006. The 2005 World Health Organization
     Reevaluation of Human and Mammalian Toxic Equivalency Factors for Dioxins and Dioxin-Like
     Compounds. Toxicol. Sciences 93(2), 223-241.
29.   Williams, R.L., Cseh, L. A review of dioxins/furans and methyl mercury in fish from the Penobscot River,
     located near Lincoln, Maine. Toxicol. Ind. Health 23, 147-153, 2007.
30.   Winters, D., Cleverly, D., Meier, K., Dupuy, A., Byrne, C., Deyrup, C., Ellis, R. Ferrario, J., Harless, R.,
     Leese, W., Lorber, M., McDaniel, D., Schaum, J., Walcott, J., 1996, A Statistical Survey of Dioxin-like
     Compounds in United States Beef: A Progress Report, Chemosphere  32: 469-478.
31.   World Health Organization, 201 la.  Toxic equivalency factors (TEFs) for dioxins and dioxin-like
     compounds.  Available at: http://www.who.int/foodsafety/chem/tefs/en/index.html[Accessed 14
     October 2012].World Health Organization, 201 Ib. 2004 Re-evaluation of human and mammalian
     toxic equivalency factors (TEFs). Available at:
     http://www.who.int/foodsafety/chem/tef_update/en/index.html [Accessed 14 October 2012].

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32.   Yeardley, R.B., Lazorchak, J.M., Pausen, S.G. Elemental Fish Tissue Contamination in Northeastern U.S.
     Lakes:  Evaluation of an Approach to Regional Assessment. 1998. Environ. Toxicol. Chem. 17 (9), 1875-
     1884.
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                                  APPENDIX C


                     Penobscot RARE Project Schedule

August 30, 2007
•  Conference call to review project schedule
•  Identify persons responsible for action items for QAPP
•  Data Quality Objectives
•  Data Usability
•  Project Action limits
•  Set up a standard time for conference calls for QAPP development.
•  Identify next steps with time frames and responsible parties

September 2007
•  Finalize Data Quality Objectives
•  Finalize Data Usability Objectives
•  Finalize Project Action Limits

October 2007
•  Finalize Sampling SOP for fish and sediment
•  Review all SOPs for analysis against Project Action Limits and identify lab concerns regarding
   any detection limit issues
•  Develop flow charts for data analysis, i.e. what each lab needs in order to conduct analysis to
   meet data quality objectives
•  Develop flow chart for chain-of-custody of samples
•  Develop flow chart of responsibilities of proj ect team members
•  Initial QA site visit

November 2007
•  Develop Sampling SOP for plant, wood duck and snapping turtle
•  Review all SOPs for analysis against Project Action Limits and identify lab concerns regarding
   any detection limit issues
•  Develop flow charts for data analysis, i.e. what each lab needs in order to conduct analysis to
   meet data quality objectives
•  Develop flow chart for chain-of-custody of samples
•  Develop flow chart of responsibilities of proj ect team members

December 2007
•  Review QAPP for other areas that need to be revised to ensure consistency with data quality
   objectives.
•  Develop and send out draft QAPP for review by December 15th, 2007

January 2008
 • Review and comment on draft QAPP
Page  93                                            Final RARE Report August 2015

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 April 2008
 • Final comments due on QAPP

 May 2008
 • Send out final QAPP for approval
 • Collect plants (Ostrich Fern at fiddlehead stage)
 • Fern samples shipped from PIN to USGS

 July 2008
 • Collection of sediment samples for contaminant analysis
 • RARE QA Officers review and conduct technical systems audit (TSA) of field sample
   collection, handling, and shipment in Maine

 July-October 2008
 • Collection offish, ducks, turtles (only able to collect 2 turtle samples)
 • Sediment samples shipped from PIN to laboratories
 • Sediment  and fern samples received by OPP/ECL

August 2008
•  RARE QA Officer at Chelmsford, MA Lab, (August 4-6, 2008)
•  Methyl mercury results received from Frontier GeoSciences (FGS) lab for sediment

September 2008
•  Methyl  mercury results received from FGS lab for fiddlehead ferns

October 2008
•  Sediment samples analyzed by OPP/ECL

October 2008-January 2009
•  Fish fillets received by EPA-NERL, homogenized, and shipped to other labs
•  Turtle meat, duck meat, and ferns received by USGS, homogenized, and shipped to labs.

March 2009
 • Fish fillet samples received by OPP/ECL

April 2009
 • Turtle samples (collected in 2008) received by OPP/ECL
 • Duck samples received by OPP/ECL
 • Fish fillet samples analyzed by OPP/ECL

 May 2009
 • Methyl mercury results received from FGS lab for turtle and duck

 July - October 2009
 • Collect additional snapping turtles from all reaches
 • Collect and ship water and sediment samples for AMES study

Page  94                                           Final RARE Report August 2015

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  •  Conduct TSA by QA Manager at the US EPA in RTF on the Salmonella (Ames) mutagenicity
    testing of the river and drinking waters.

 September - October 2009
 •  Collect Medicinal Plants
 •  Attempt to collect wood ducks from BBS reach or adjacent area.

  October 2009
  •  Fern samples were analyzed by OPP/ECL
  •  Duck samples were analyzed by OPP/ECL
  •  Turtle samples (collected in 2008) were analyzed by OPP/ECL

 November 2009
 •  Analytical Data Reports from Laboratories sent to US EPA data validator
 •  Data validation conducted by US EPA

 December 2009
 •  Validated data sent from data validator to EPA-ORD.

 February 2010
 •  Medicinal Plants and Turtle samples (collected in 2009) shipped from PIN to USGS

 March 2010
 •   Data Validation memos issued

 July 2010
 •  Turtle meat (collected in 2009) and medicinal plants received by USGS, homogenized, and
    shipped to lab
 •  Turtle samples collected from 2009 received by OPP/ECL
 •  Medicinal plant samples received by OPP/ECL

 August 2010
  •  Turtle samples collected from 2009 analyzed by OPP/ECL

 September 2010
 •  Medicinal plant samples analyzed by OPP/ECL

October 2010
 •  Analysis completed October 2010
 •  Final data audit was conducted in Maine
 •  Project team met at PIN to evaluate available data

 February 2011
 •  ORD Draft Report issued ( ORD lead retired)

 2011-2013
 •  Region 1 co-lead development of Draft Report

 Page  95                                           Final RARE Report August 2015

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•  Development of Peer Review Charge questions with team

April
 2013-May 2013
•  Peer Review

May 2013 - January 2014
•  Peer Review edits incorporated and responses to comments developed.

January 2014
•  Draft Report presented to Penobscot Tribal Council for review and comments

October 2014
•  Internal EPA Region 1  (Rl) review.
•  Coordination with Rl IT to develop final report with a CD for distribution

October 2014-April 2015
•  Incorporation of all comments
•  Development of table of contents for CD

May-August 2015
•  Coordination with Rl and ORD for approval and presentation of final RARE report
•  Obtained EPA publication number for final RARE report
•  Incorporated final changes requested by ORD
Page  96                                           Final RARE Report August 2015

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                                  APPENDIX D
            Personnel Associated with the RARE Study

United States Environmental Protection Agency
Robert Hillger, US EPA Senior Science Advisor
RARE Program Coordinator
New England Regional Laboratory
11 Technology Dr. North Chelmsford, Mass. 01863
(617) 918-8660; Hillger.Robert@epa.gov

Valerie Marshall, US EPA Region 1
RARE Project Role: Project Leader and QAPP Approver
EPA Boston, MA
(617) 918-1674; Marshall.Valerie@epa.gov

Janet J. Diliberto, Research Biologist
RARE Project Role: Project Leader and QAPP Approver
USEPA/ORD/NHEERL/ISTD; Office B458
109 TW Alexander Drive, Mail Drop B105-01
Research Triangle Park, NC 27711
(919)541-7921; FAX: (919) 541-9464: Diliberto.Janet@epa.gov

Thomas Hughes, US EPA, QA and Records Manager
RARE Project Role: QA Officer and Records Manager and QAPP Approver
EPA/ORD/NHEERL/RCU
109 TW Alexander Drive, Mail Drop B105-01
Research Triangle Park, NC 27711
(919) 541-7644; Hughes.Thomas@epa.gov

David M. DeMarini, Toxicologist
RARE Project Role: Mutagenicity Study
USEPA/ORD/NHEERL/ISTD
109 TW Alexander Drive, Mail Drop B105-03
Research Triangle Park, NC 27711
(919) 541-1510; Demarini.David@epa.gov

Richard H. Sugatt, Environmental Scientist
RARE Project Role, preliminary risk assessment
EPA Boston, MA
(617) 918-1415; Sugatt.Rick@epa.gov

Sarah H. Warren
RARE Project Role: Mutagenicity Study
USEPA/ORD/NHEERL/ISTD
109 TW Alexander Drive, Mail Drop B105-03
Research Triangle Park, NC 27711
(919) 541-0975; Warren.Sarah@epa.gov
Page  97                                            Final RARE Report August 2015

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Adam Swank
RARE Project Role: Mutagenicity Study
USEPA/ORD/NHEERL/RCU
109 TW Alexander Drive, Mail Drop B105-01
Research Triangle Park, NC 27711
(919) 541-0614; Swank.Adam@epa.gov

Steve DiMattei, US EPA, QA Chemist Region 1
RARE Project Role: QA Officer, QAPP Approver, and Region 1 QA contact
New England Regional Laboratory
11 Technology Drive, North Chelmsford, MA 01863
(617) 918-8369; dimattei.steveigiepa.gov

Dave McDonald, US EPA, Biology Laboratory Manager
RARE Project Role: Biology QA Officer for US EPA NERL, QAPP Reviewer
New England Regional Laboratory
11 Technology Dr. N. Chelmsford, MA 01863
(617) 918-8609; FAX (617) 918-8509; mcdonald.dave@epa.gov

Joseph Ferrario, US EPA, Lab Director/ Dioxin Team Leader
RARE Project Role: Leader of OPP/Stennis dioxins, furans, WHO PCBs; QAPP Reviewer
Lab Contact for RARE Project
EPA/Office of Pesticide Programs, Biological and Economic Analysis Division, Environmental Chemistry
Branch, NASA/SSC Building 1105,
Stennis Space Center, MS 39529-6000
(228) 688-3171/3212; ferrario.joseph@epa.gov

Christian Byrne, US EPA-OPP Quality Assurance Officer
RARE Project Role: OPP Data Approval; QAPP Reviewer
EPA/OPP/BEAD/ECB, NASA/SSC Bldg 1105,
Stennis Space Center, MS 39529-6000
(228) 688-3213; Byrne.Christian@epa.gov

Craig Vigo, Mass Spectrometrist
RARE Project Role: Chemical Analysis
EPA/OPP/BEAD/ECB;Stennis Space Center, MS 39529-6000
(228) 688-1229; Vigo.Craig@epa.gov

Tripp Boone, Safety Officer/ Sample Prep Coordinator
RARE Project Role: Sample Custodian for samples shipped to US EPA-OPP Stennis.
EPA/OPP/BEAD/ECB, Stennis Space Center, MS 39529-6000
(228) 688-2242;  Boone.Tripp@epa.gov

Stanley Mecomber
RARE Project Role: Sample Custodian
EPA/OPP/BEAD/ECB,Stennis Space Center, MS 39529-6000
(228)688-3172/3212; mecomber.stanley@epa.gov

Steve Stodola, US EPA, QA Chemist
RARE Project Role: Data validation.
US EPA -NERL, OEME, 11 Technology Drive, N. Chelmsford, MA 01863
(617)918-8634;   stodola.steve@epa.gov

Page  98                                            Final RARE Report August 2015

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Alan VanArsdale, Direct Mercury Analyzer (DMA)
RARE Project Role:  Responsible for scheduling and analyzing samples for mercury analysis using the Direct
Mercury Analyzer (DMA).
New England Regional Laboratory
11 Technology Dr. North Chelmsford, MA 01863
(617) 918-8610; vanarsdale.alan@epa.gov

James M. Lazorchak
Aquatic Ecologist/Toxicologist
Manager, AAALAC Certified Aquatic Research Facility
RARE Project Role: Responsible for filleting fish samples
ORD NERL EBRD
U.S. EPA
26 W. Martin Luther King Dr
Cincinnati, OH 45268
Phone 513 569 7076
cell    513 550 1537
Fax    5135697438
Email:  Lazorchak.jim@epa.gov
web:  http://www.epa.gov/eerd/

ESAT
ESAT or Technician from Narragansett Lab
RARE Project Role:  Fish processing with field team
Wannalancit Mills Technology Park, 175 Cabot Street, Suite 415, Lowell, MA 01854
Phone: 978-275-9730

TechLaw, Inc.
RARE Project Role:  Data Validation
Wannalancit Mills Technology Park, 175 Cabot Street, Suite 415, Lowell, MA 01854
Phone: 978-275-9730

Penobscot Indian Nation Department of Natural Resources
Daniel Kusnierz, PIN-DNR, Water Resources Program Manager
RARE Project Role: RARE PIN-DNR Leader; Assists in field sampling; QAPP Review
Penobscot Indian Nation - DNR, 12 Wabanaki Way, Indian Island, Old Town, ME 04468
(207) 817-7361 or (207) 827-7776 ext. 7361; Dan.Kusnierz@penobscotnation.org

Jason Mitchell, PIN DNR, Water Resources Field Coordinator
RARE Project Role: Assist with field sampling
Penobscot Indian Nation - DNR; 12 Wabanaki Way, Indian Island
Old Town, ME 04468,  supervisor: Dan Kusnierz;
(207)817-7381;  Jason.Mitchell@penobscotnation.org

Jan Paul, PIN DNR Water Resources Field/Lab Technician;
RARE Project Role: Assist with field sampling
Penobscot Indian Nation - DNR; 12 Wabanaki Way, Indian Island; Old Town, ME 04468
Supervisor: Dan Kusnierz
(207)817-7382;  Jan.Paul@penobscotnation.org
Page  99                                              Final RARE Report August 2015

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Michele Attean, PIN DNR Water Resources Seasonal Field/Lab Technician;
RARE Project Role: Assist with field sampling
Penobscot Indian Nation - 12 Wabanaki Way, Indian Island, Old Town, ME 04468
Supervisor: Dan Kusnierz

Jason Sockbeson, PIN DNR Water Resources Seasonal Field/Lab Technician;
RARE Project Role: Assist with field sampling
Penobscot Indian Nation - 12 Wabanaki Way, Indian Island, Old Town, ME 04468
Supervisor: Dan Kusnierz

Joe Dana, PIN DNR Wildlife Technician;
RARE Project Role: Assist with field sampling
Penobscot Indian Nation - 12 Wabanaki Way, Indian Island, Old Town, ME 04468
Supervisor: Kristin Peet

Kristin Peet, PIN DNR Wildlife Biologist
RARE Project Role: Assist with collecting ducks and other sampling
Penobscot Indian Nation - DNR; 12 Wabanaki Way, Indian Island; Old Town, ME 04468
Supervisor: John Banks; (207)817-7363; Kristin.Peet@penobscotnation.org

Frontier GeoSciences, Inc.
Matthew Gomes,
RARE Project Role: FGS Project Manager
Frontier GeoSciences, Inc.
414 Pontius Ave. N
Seattle, WA 98109
(206)622-6960 x 1449; mattg@frontiergeosciences .com

Patrick Garcia Strickland
RARE Project Role: FGS Lab Manager
Frontier GeoSciences, Inc.
414 Pontius Ave. N
Seattle, WA 98109
(206)622-6960 x 1428; patricks@frontiergeosciences.com

Ryan Nelson
RARE Project Role: FGS Lab Mercury Group Leader
Frontier GeoSciences, Inc.
414 Pontius Ave. N
Seattle, WA 98109
(206)622-6960 x 2012; rvann@frontiergeosciences.com

Kristina Spadafora
RARE Project Role: FGS QA Officer
Frontier GeoSciences, Inc.
414 Pontius Ave. N
Seattle, WA 98109
(206)622-6960 x 1423; kristinas@frontiergeosciences.com
Page   100                                                     ;. :-,   \              2015

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United States Geological Survey
Robert W. Dudley, USGS, Hydrologist
RARE Project Role: Field Sampling Leader; QAPP Review;
USGS Maine Water Science Center, Augusta ME
(207) 622-8201 ext. 115; rwdudley@usgs.gov

Charles Culbertson, Alternate to Rob Dudley
USGS Maine Water Science Center
196WhittenRoad
Augusta, ME 04330
(207) 622-8201 ext. 127

James Caldwell, Alternate to Rob Dudley
USGS Maine Water Science Center
196WhittenRoad
Augusta, ME 04330
(207) 622-8201 ext. 107

Robert M. Lent, USGS, Director of Maine Water Science Center
RARE Project Role: USGS Field Sampling Project Manager, review sampling
Method SOPs, QAPP Review
USGS Maine Water Science Center, Augusta ME
(207) 622-8201 ext. 102; rmlent@usgs.gov

Carl E. Orazio, PhD. USGS-CERC Branch Chief Environmental Chemistry
RARE Project Role: USGS Project Officer (CERC/USGS US EPA TAG); QAPP Preparation; Review Analytical
Methods SOPs
CERC USGS Lab Contact: Congener-specific PCB and Mercury analyses.
USGS Columbia Environmental Research Center (CERC)
4200 New Haven Road, Columbia, MO  65201;
(573) 876-1823;  corazio@usgs.gov

Robert Gale, PhD. USGS/CERC, Leader Environmental Fate and Dynamics
RARE Project Role: Supervisor of congener-specific PCB analysis
USGS, 4200 New Haven Road, Columbia, MO 65201;
(573) 875-5399 (Supervisor: Carl Orazio)

Kathy Echols, PhD. USGS/CERC, Leader Complex Contaminant Mixtures
RARE Project Role: Review of congener-specific PCB analysis
USGS, 4200 New Haven Road, Columbia, MO 65201;
(573) 875-5399 (Supervisor: Carl Orazio)

John Meadows, USGS/CERC, Dioxin and PCB Chemist
RARE Project Role: Conduct congener-specific PCB analysis.
USGS, 4200 New Haven Road, Columbia, MO 65201
(573) 875-5399 (Supervisor: Carl Orazio)

George Tegerdine, USGS/CERC, PCB congener analysis Technician,
RARE Project Role: Conduct congener-specific PCB GC analysis.
USGS, 4200 New Haven Road, Columbia, MO 65201
(573) 875-5399 (supervisor: Carl Orazio)
Page  101                                           Final'-"-

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Tom May, USGS/CERC Leader Toxic Element Research,
RARE Project Role: Supervisor of Total-Mercury Analysis.
4200 New Haven Road, Columbia, MO 65201; USGS/CERC
(573) 876-1858; (supervisor: Carl Orazio)

William Brumbaugh, USGS/CERC Research Chemist,
RARE Project Role: Mercury Analysis expert and methods reviewer
USGS, 4200 New Haven Road, Columbia, MO 65201
(573) 876-1857; (supervisor: Carl Orazio)

Paul Peterman, USGS/CERC Trace Organic Contaminants Research Chemist,
RARE Project Role: Dioxin and PCB Analysis expert and methods reviewer.
USGS, 4200 New Haven Road, Columbia, MO 65201
(573) 875-5399;  ppeterman@usgs.gov (supervisor: Carl Orazio)

Kevin Feltz, USGS/CERC Trace Organic Contaminants Chemist,
RARE Project Role: Dioxin and PCB Analysis expert and methods reviewer.
USGS, 4200 New Haven Road, Columbia, MO 65201
(573) 875-5399 (supervisor: Carl Orazio)

Michael Walther, USGS/CERC Technician,
RARE Project Role: Total-Mercury Analysis.
USGS, 4200 New Haven Road, Columbia, MO 65201
(575) 875-5399 (supervisor: Carl Orazio)

Jesse Arms, USGS Technician, Sample Receiving
RARE Project Role: Sample Receiving
USGS, 4200 New Haven Road, Columbia, MO 65201
(575) 876-1856 (supervisor: Carl Orazio 876-1823)

Paul Heine, USGS CERC QA Officer
USGS, 4200 New Haven Road, Columbia, MO 65201
(573)876-1815;  pheine@usgs.gov

Adria A. Elskus, Ph.D./USGS/LSC/CAFRL
Aquatic Toxicology
RARE Project Role: Alternate for Carl Orazio
5751 Murray Hall
University of Maine
Orono, ME 04469-5751
PH: 207/5 81-25 79
aelskus@usgs.gov

Agency for Toxic Substances and Disease Registry (ATSDR)
Lynn Wilder, PhD
RARE Project Role: RARE ATSDR Lead, QAPP Approver
Division of Community Health Investigations
4770 Buford Highway, N.E. MS F59, Atlanta, GA 30341
770-488-3688
Ixw2@cdc.gov
Page  102                                           Final'1."-

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Gary D. Perlman MPH, RS, EMT-B
RARE Project Role, Commander, US Public Health Service
ATSDR Region 1 (New England)
1 Congress St. Suite 1100, Mail Code HBT
Boston, MA 02114-2023
Phone: (617) 918-1492; Cell: (617)842-1560; Fax: (617)918-1494
Email: gap6@CDC.GOV orperlman.gary@epa.gov

Katherine Pugh, Environmental Health Scientist,
RARE Project Role: evaluating the ingestion pathway for the watershed species
ATSDR Division of Community Health Investigations
4770 Buford Highway, N.E. MS F59, Atlanta, GA  30341
770-488-0765
krh2@cdc.gov

Annabelle Allison,
NCEH/ATSDR Office of Tribal Affairs Coordinator.
National Center for Environmental Health (NCEH)
U.S. Centers for Disease Control and Prevention
4770 Buford Highway, MS F-61Atlanta GA 30341-3717
770-488-3991
AAllison@cdc.gov

Bureau of Indian Affairs
Kelly Gupton
RARE Project Role: Representative of BIA
Water Resources, Bureau of Indian Affairs
Eastern Regional Office, 545 Marriott Drive, Suite 700
Nashville, TN 37214
(615)  564-6838; FAX (615)564-6571

United State Fish & Wildlife  Service
Steve  Mierzykowski, Senior Fish & Wildlife Biologist
U.S. Fish & Wildlife Service
17 Godfrey Drive, Suite #2
Orono, Maine 04473
Office: (207) 866-3344 ext. 112
Cell: (207) 944-3007
Fax:(207)866-3351
steve  mierzykowski@fws .gov

National Institute of Environmental Health Sciences
*Linda Birnbaum,  Director National Institutes of Health (NIH), and National Toxicology Program
P.O. Box 12233
Mail Drop B2-01
Research Triangle  Park, North Carolina 27709
Tel (919) 541-3201
Fax (919) 541-2260
(*Linda participated in this study while working for US EPA/ ORD)
Page  103                                            Final'1."-

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                                          APPENDIX E
                                  RARE Quality Assurance Statement

                                        U.S. EPA ORD, RTF, NC
              "Developing Exposure Concentrations for Regional Cultural Tribal Exposure Assessment,
                                         Penobscot River, Maine"

                                              Page 1 of2

           The study "Developing Exposure Concentrations for Regional Cultural Tribal Exposure
           Assessment Penobscot River, Maine" was conducted by the National Health and Environmental
           Effects Research Laboratory (NHEERL), Office of Research and Development, U.S.
           Environmental Protection Agency, Research Triangle Park, NC, in compliance with NHEERL
           and ORD QA Guidelines. These audits were conducted by Tom Hughes, US EPA QA and
           Records Manager and Program QAM (PQAM) for this ORD RARE, and Janet Diliberto, Co-
           Principle Investigator. This RARE study was conducted in collaboration with Region 1 of the
           US EPA. Old Town, Maine is near Indian Island, the home of the Penobscot Indian Nation.
           Critical phases in the study were audited.
           Date of Inspection

           October 15-17,2007



           July 21-24, 2008




           October 6-9, 2008


           Nov 2-3, 2009
           October 12-14, 2010
Type              Items Inspected

Site Visit,           Inspect reaches of the Penobscot
Old Town, ME      River; Meet research team;
                   Have discussions on QAPP

TSA, Old Town,     Observe sampling offish from the
ME                Penobscot River; Observe shipping
                   of fish; Audit records; Meet with
                   team members

TSA, North         Audit Mercury Lab for RARE
Chelmsford, MA     project; Meet with team members

TSA, RTP, NC      Audit of Ames/Salmonella
                   Mutagenicity results from samples of
                   river water, drinking water and
                   sediment from the Penobscot River
                   Conducted by Barbara Collins,
                   QAM, ISTD, US EPA, RTP, NC

Surveillance         Inspect records of Penobscot Indian
Old Town, ME      Laboratory for RARE project.
           TSA = technical systems audit
Page  104
                     Final RARE Report August 2015

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                                    RARE Quality Assurance Statement

                                          U.S. EPA ORD, RTF, NC
                 "Developing Exposure Concentrations for Regional Cultural Tribal Exposure Assessment,
                                           Penobscot River, Maine"

                                                Page 2 of2
              The Program Quality Assurance Manager (PQAM) of this ORD RARE program and the Co-Pi
              of this RARE Program have determined by the above review process that the conduct of this
              program was in compliance with US EPA quality requirements and the operating procedures and
              study Quality Assurance Project Plan (No.: ORD RARE- Region 1- Diliberto.Bataille/2008-01-
              rOO).  Furthermore, the results accurately reflect the raw data obtained during the course of the
              study.
                    /^•Ct-q'K-l^	
Thomas J. Hughes^USEPA RARE QA and Records Manager
                                                                    Date
             Valerie Marshall, USEPA RARE Co-Principle Investigator
                                                                      £-\\  -
                                                      Date
Page  105
                                                Final RARE Report August 2015

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                     Appendix F
Risk of Each Biota Type in Each Reach at the Inland Non-Anadromous Tribal
Ingestion Rate
Reach


1
1
1
2
2
2
3
3
3
4
4
4
5
5
5
6
6
6
2
2
2
3
3
3
4
4
4
6
6
6
1
1
1
4
4
Biota
Type

CP
CP
CP
CP
CP
CP
CP
CP
CP
CP
CP
CP
CP
CP
CP
CP
CP
CP
YP
YP
YP
YP
YP
YP
YP
YP
YP
YP
YP
YP
WP
WP
WP
WP
WP
Chemical


PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Cone, in
Biota
(mg/kg)

ND
4.24E-08
4.32E-01
ND
5.93E-08
8.09E-01
ND
3.99E-08
8.67E-01
ND
3.78E-08
3.16E-01
ND
1.26E-07
5.88E-01
ND
5.56E-08
5.44E-01
ND
3.83E-07
5.36E-01
ND
6.41E-08
4.16E-01
ND
3.43E-08
1.46E-01
ND
1.54E-08
2.84E-01
ND
3.83E-07
5.36E-01
ND
4.02E-07
Tribal
Ingestion
Rate
(g/day)
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
Risk Based Concentration

HQ=1
(mg/kg)
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
CR=1E-
06
(mg/kg)
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
Risk in Biota

HQ


2E-01
2E+01

3E-01
3E+01

2E-01
3E+01

2E-01
1E+01

7E-01
2E+01

3E-01
2E+01

2E+00
2E+01

4E-01
2E+01

2E-01
6E+00

9E-02
1E+01

2E+00
2E+01

2E+00
CR


9E-06


IE-OS


9E-06


8E-06


3E-05


IE-OS


8E-05


IE-OS


7E-06


3E-06


8E-05


9E-05
Page  106
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4
5
5
5
6
1
2
2
2
3
3
3
4
4
4
5
5
5
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6
1
1
1
2
2
2
3
3
3
4
4
4
5
5
5
6
6
6
Biota
Type



WP
WP
WP
WP
WP
SMB
SMB
SMB
SMB
SMB
SMB
SMB
SMB
SMB
SMB
SMB
SMB
SMB
SMB
SMB
SMB
BB
BB
BB
BB
BB
BB
BB
BB
BB
BB
BB
BB
BB
BB
BB
BB
BB
BB
Chemical




Hg
PCB
TEC-
HE
PCB
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
Cone, in
Biota
(mg/kg)

-
4.67E-01
ND
8.12E-07
6.27E-01
ND
8.03E-01
5.05E-04
1.67E-07
9.45E-01
6.86E-04
1.32E-07
9.79E-01
1.25E-03
2.44E-07
9.65E-01
1.10E-03
1.83E-07
9.61E-01
8.99E-04
2.11E-07
8.09E-01
ND
3.37E-07
2.90E-01
ND
1.50E-07
4.23E-01
ND
2.97E-07
2.52E-01
ND
3.15E-07
1.80E-01
ND
7.27E-07
4.16E-01
ND
2.09E-07
1.35E-01
Tribal
Ingestion

Rate
(g/day)
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
Risk Based Concentration


HQ=1
(mg/kg)
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
CR=1E-
06
(mg/kg)
NA
2.98E-04
4.58E-09
NA
2.98E-04
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
Risk in Biota


HQ

2E+01

5E+00
2E+01

3E+01
1E-01
9E-01
4E+01
1E-01
7E-01
4E+01
2E-01
1E+00
4E+01
2E-01
1E+00
4E+01
2E-01
1E+00
3E+01

2E+00
1E+01

8E-01
2E+01

2E+00
1E+01

2E+00
7E+00

4E+00
2E+01

1E+00
5E+00

CR



2E-04


OE+00
2E-06
4E-05
4E-05
2E-06
3E-05
3E-05
4E-06
5E-05
6E-05
4E-06
4E-05
4E-05
3E-06
5E-05
5E-05

7E-05


3E-05


6E-05


7E-05


2E-04


5E-05

Page  107
Final RARE Report August 2015

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2
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1
1
2
2
2
3
3
3
4
4
4
1
1
1
2
2
2
3
3
3
4
4
Biota
Type
AE
AE
AE
AE
AE
AE
AE
AE
AE
AE
AE
AE
AE
WD
WD
WD
WD
WD
WD
WD
WD
WD
WD
WD
WD
FOF
FOF
FOF
FOF
FOF
FOF
FOF
FOF
FOF
FOF
FOF
Chemical
PCB
TEQ
Hg
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Cone, in
Biota
(mg/kg)
ND
1.18E-06
6.66E-01
6.35E-01
ND
1.40E-06
3.37E-01
ND
5.45E-06
7.39E-01
ND
4.61E-07
2.14E-01
1.16E-04
2.87E-07
4.79E-02
5.01E-03
4.26E-07
2.65E-02
2.44E-03
4.54E-07
2.41E-02
4.05E-03
1.08E-06
1.68E-02
6.12E-04
3.21E-10
1.30E-03
1.15E-03
1.79E-10
8.00E-04
ND
4.42E-09
8.00E-04
3.22E-04
ND
Tribal
Ingestion
Rate
(g/day)
286
286
286
286
286
286
286
286
286
286
286
286
286
70
70
70
70
70
70
70
70
70
70
70
70
133
133
133
133
133
133
133
133
133
133
133
Risk Based Concentration
HQ=1
(mg/kg)
5.10E-03
1.79E-07
2.55E-02
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
2.09E-02
7.30E-07
1.04E-01
2.09E-02
7.30E-07
1.04E-01
2.09E-02
7.30E-07
1.04E-01
2.09E-02
7.30E-07
1.04E-01
1.10E-02
3.84E-07
5.49E-02
1.10E-02
3.84E-06
5.49E-02
1.10E-02
3.84E-07
5.49E-02
1.10E-02
3.84E-07
CR=1E-
06
(mg/kg)
2.98E-04
4.58E-09
NA
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
1.22E-03
1.87E-08
NA
1.22E-03
1.87E-08
NA
1.22E-03
1.87E-08
NA
1.22E-03
1.87E-08
NA
6.40E-04
9.85E-09
NA
6.40E-04
9.85E-08
NA
6.40E-04
9.85E-09
NA
6.40E-04
9.85E-09
Risk in Biota
HQ
7E+00
3E+01
2E+01
8E+00
1E+01
3E+01
3E+01
3E+00
8E+00
6E-03
4E-01
5E-01
2E-01
6E-01
3E-01
1E-01
6E-01
2E-01
2E-01
1E+00
2E-01
6E-02
8E-04
2E-02
1E-01
5E-04
1E-02
1E-02
1E-02
3E-02
CR
3E-04

3E-04

1E-03
1E-04
1E-07
2E-05
2E-05
4E-06
2E-05
3E-05
2E-06
2E-05
3E-05
3E-06
6E-05
6E-05
1E-06
3E-08
1E-06
2E-06
2E-08
2E-06
4E-07
5E-07
Page  108
Final RARE Report August 2015

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6
3
3
3
4
4
4
5
5
6
6
1
1
1
3
3
3
4
4
4
5
5
5
6
6
6
Biota
Type




FOF
MP
MP
MP
MP
MP
MP
MP
MP
MP
MP
ST
ST
ST
ST
ST
ST
ST
ST
ST
ST
ST
ST
ST
ST
ST
Chemical





Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
PCB
TEQ
Hg
Cone, in
Biota

(mg/kg)

-
8.00E-04
ND
6.40E-08
6.92E-03
ND
9.02E-08
8.61E-03
ND
2.40E-08
3.60E-08
2.92E-03
ND
4.86E-06
9.63E-01
ND
7.49E-07
5.69E-01
2.14E-02
2.04E-06
6.05E-01
ND
2.80E-06
1.046E+00
1.70E-04
1.44E-07
2.77E-01
Tribal
Ingestion


Rate
(g/day)
133
133
133
133
133
133
133
133
133
133
133
286
286
286
286
286
286
286
286
286
286
286
286
286
286
286
Risk Based
Concentration


HQ=1
(mg/kg)
5.49E-02
1.10E-02
3.84E-07
5.49E-02
1.10E-02
3.84E-07
5.49E-02
1.10E-02
3.84E-07
3.84E-06
5.49E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
5.10E-03
1.79E-07
2.55E-02
Risk in
Biota

CR=1E-
06
(mg/kg)
NA
6.40E-04
9.85E-09
NA
6.40E-04
9.85E-09
NA
6.40E-04
9.85E-09
9.85E-08
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA
2.98E-04
4.58E-09
NA


1E-01

HQ

1E-02

2E-01
1E-01

2E-01
2E-01

6E-02
9E-02
5E-02

3E+01
4E+01

4E+00
2E+01
4E+00
1E+01
2E+01

2E+01
4E+01
3E-02
8E-01
1E+01




CR



6E-06


9E-06


2E-06
4E-06


1E-03


2E-04

7E-05
4E-04
5E-04

6E-04

6E-07
3E-05
3E-05
CP = Chain Pickerel
YP = Yellow Perch
WP = White Perch
SMB= Smallmouth Bass
BB = Brown Bullhead
AE = American Eel
WD = Wood duck
FOF = Fiddlehead Ostrich Fern
MP = Medicinal Plant
ST = Snapping Turtle
CR = Cancer Risk
HQ = Hazard Quotient
HQ = RBC for HQ=1/Concentration in Biota
CR = BC for CR= lE-06/Concentration in Biota x 1E-06
ND = Non-Detect Risk values are rounded to the nearest whole number.
PCB =  Polychlorinated Biphenyls
TEQ = dioxin Toxic Equivalents
Hg = Mercury
Number in gray is sum of cancer risks for PCB and TEQ
Page  109
                        Final RARE Report August 2015

-------
Computer Printout from EPA Regional Screening Level Calculator
                (http://epa-prgs.ornl.gov/cgi-bin/chemicals/csl search)
Note: This printout documents the chemicals, exposure parameters and toxicity factors
that were entered into the calculator to calculate risk-based concentrations for Hazard
Index =1 and cancer risk of 1E-06 for ingestion of food.
Page  110                                      Final RARE Report August 2015

-------
                Site-specific
                Fish Equation Inputs for Fish
                 Variable
                 TR (target cancer risk) unitless
                 AT (averaging time)
                 EF, (exposure frequency) dayS/yr
                 ED, (exposure duration) yr
                 LT (lifetime) yr
                 BW, (body weight) kg
                 (RFa (fish consumption rate) mg/cJay 133000
                Output generated  05NOV2012:13:33:44
                Site-specific
                Ptsh Risk-Based Screening Levels (RSL) for Pish
                CJuCancer. nc«Monc3(«er. ca1 (Where re SL < 100 * C» SC),
                ca" |Wh«r» nc SL < 10 x c* SU.
                nui^L txc««d» ctlling Ilinll ISM UMM'* Guide), sat=SL avccrdt rial

                                       Ingestion SF
                   Chemical
                                      Ingestion of              Ingestion
                         Chronic          Fish     Carcinogenic   of Fish  Noncarcinogenic Screening
 CAS              SFO    RfO     RfD SL - TR=1.0E-6 SL - TR=1.0E-6 SL - HQ=1     SL - Hl=1       Level
Number  
-------
                      Site-specific
                      Fish Equation Inputs for Fish
                       Variable
                       TR (target cancer risk) unitless
                       AT (averaging time)
                       EF, (exposure frequency) days/yr
                       ED, (exposure duration) yr
                       LT (lifetime) yr
                       BW, (body weight) kg
                       RFa (fish consumption rate) mg/day 70000
                      Output generated  05NOV20l2:13;28:02
                                                                          Ingestion of               tngestion
                                                             Chronic          Fish      Carcinogenic   of Fish  Noncarcinogenic  Screen
                                     CAS             .. SFO   RfD    RfD SL - TR=1.0E-6 SL - TR*1.0E-6  SL • HQ=1    SL - Hl=1       Level
                                    Number  (mg/kg-day)* Ref (mg/kg-day) Ref    (mg/kg)       (mg/kg)     (mg/kg)      (mg/kg)       (mg/kg)
                      Site-specific
                      Fish Risk-eased Screening Levels (RSL) for Fish
                      ca=C3ficer. nc*None*new. ca' IWtier* nc SI. « 100 » ca SL},
                      CJ" (Where nc SL < 10 x c» SL).
                      m«X*SL «KCMds tciliny llmil ls«« Uwf'S Guk)*). Ml=SL «C*«f9
                       TCDD.2.3.7.B.ll7d6-01-6 !
                       Methyl Mercury 22967-92-6            	  1 .OOE-Q4
                       Aroclor12S4! 11097-69-11   2.00E*00  I S  I  2'• -. <"•••
                       Output generated  05NOV2012:13:28:02
Page    112
Final RARE Report August 2015

-------
                      Site-specific
                      Fish Equation Inputs for Fish
                      Variable                    value
                      TR (target cancer risk) unittess    1 .OE-6
                      AT (averaging time)            365
                      EP, (exposure frequency) days/yr  350
                      £Dr (exposure duration) yr       30
                      I_T (lifetime) yr                70
                      3W, (body wefght) kg          70
                      IRF (fish consumption rate) mg/day 286000
                      Output generated  05NOV2012:13:35:35
                      Site-specific
                      Fish Risk-Based Screening Levels (RSL) for Fish
                                =Noncanc.»r. c.i* (Wher* nc SL < 100 » cjt SU),
                         (Where ne SL < 10 n C» SL).
                              eed! CAlMng limit ise* User's Guide). &at=SL exceeds
                                                                        Ingestion of              Ingesbon
                                                            Chronic         Fish     Carcinogenic   of Fish  Noncarcinogenic Screening
                                     CAS              SfO    RfD    RfO SL • TR=1.0£-6 SL - TR=1.0E-6 SL - HQ-1    SL - Hl*1      Level
                                   Number  (mg/kg-day)' Ref (mg/kg-day) Ref   (mfl/kg)      (mg/kg)     (mg/kg)     (mg/kg)      (mgJkg)
                      Outpul generated 05NOV2012:13:36:35

Page    113
Final RARE Report August 2015

-------
                               APPENDIX G
Penobscot RARE Peer Review Panel
Expertise
Tribal Risk
Assessment
State Health
Assessor, CT
Mutagenicity testing
Tribal Risk
Assessment
Green Chemist,
EPAR1
Toxicologist, Maine
CDC
Contact Information
Barbara Harper, PhD, DABT
Program Manager, Environmental Health
Department of Science and Engineering
Confederated Tribes of the Umatilla Indian Reservation
Pendleton, Oregon
Phone:(541)429-7950
BarbaraHarper@ctuir. com
Brian Toal, M.S.P.H.
28 Lawrence Avenue
Avon, Connecticut 06001
Phone: 860-5097742 (W)
brian.toal@po. state, ct.us
George M. Woodall, PhD
National Center for Environmental Assessment
Office of Research and Development
U.S. Environmental Protection Agency
MD B243-01, Research Triangle Park, NC 2771 1
Office Phone: (919) 541-3896
Email: woodall.george@epa.gov
Jamie Donatuto, PhD
Swinomish Indian Tribal Community
Department of Environmental Protection
1 1430 Moorage Way,
La Conner, WA 98257
office: (360) 466-1 532
j donatuto@swinomish.nsn.us
Nicholas Anastas, Ph.D., M.S.
USEPA Region 1
Office of Environmental Stewardship
5 Post Office Square
Boston, MA 02 109
Phone:(617)918-1177
Pamela Wadman
Maine Center for Disease Control and Prevention
1 1 State House Station
Augusta, ME 04333
Phone: (207) 287-3223
Page  114
Final RARE Report August 2015

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Penobscot RARE Peer Review Panel
Expertise
Research Chemist,
ORDR1
Research
Hydrologist, VSGS
Toxicologist, EPA
Region 8
Toxicologist, EPA
Region 9
Contact Information
Richard Pruell, PhD
USEPA ORD NHEERL
Atlantic Ecology Division
27 Tarzwell Drive
Narragansett, RI 02882
pruell.richard@epa.gov
Phone: (401)782-3091
Timothy Reilly
US Geological Survey
3450 Princeton Pike, Suite 110
Lawrenceville, NJ 08648
(609) 771-3962
Wendy Pott O'Brien, DVM, PhD, DABT
Toxicologist, Ecosystems Protection and Remediation
USEPA Region 8
1595 Wynkoop Street
Denver, CO 80202
Phone: 303. 3 12.6712
FAX: 303.312.7151
Daniel Stralka, PhD
Superfund Technical Support Section
USEPA Region 9
75 Hawthorne Street
San Francisco, CA 3901
Phone:(415)972-3048
Page  115
Final RARE Report August 2015

-------