United States
Environmental Protection
Agency
Great Lakes
National Program Office
77 West Jackson Boulevard
Chicago, Illinois 60604
EPA-905-R-95-017t-~-
September 1995
SERA The Effects of
Great Lakes Contaminants
On Human Health
Report to Congress
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FOREWORD
The Great Lakes basin comprises one-fifth of the total fresh water on the earth's
surface; it is a valuable natural resource for both the United States and Canada.
For over 200 years, the Great Lakes basin has been used as a resource for
industry, agriculture, shipping, and recreation. Approximately 10 percent of the
United States population lives within the Great Lakes basin. Researchers have
identified 362 contaminants in the water, sediment, and biota in quantifiable
amounts, approximately one-third have been evaluated for their potential toxic
effects. Many of these chemicals have known adverse effects on humans.
Critical Great Lakes pollutants identified by the International Joint Commission
are polychlorinated biphenyl (PCBs), dichlorodiphenyl trichloroethane (DDT),
dieldrin, toxaphene, mirex, methylmercury, benzo[a]pyrene, hexachlorobenzene
(HCB), furans, dioxins, and aklylated lead. All eleven substances tend to
accumulate in aquatic organisms eaten by Great Lakes fish. Because of the
persistence and widespread contamination of these Great Lakes pollutants in the
environment, toxic effects in wildlife have been demonstrated and results from
epidemiological investigations suggest that adverse human health effects, (i.e.,
reproductive, developmental, behavioral, neurologic, endocrinologic, and im-
munologic) are associated with exposure to Great Lakes pollutants.
Exposure assessment studies indicate that humans are the final biological
receptors for many toxic substances. There is limited information on the effects
associated with the consumption of Great Lakes fish. Given the implications of
the association between contaminants in the Great Lakes and adverse human
health outcomes, further research is necessary to illuminate the relationship
between exposure, contaminant levels in human biological tissues and fluids, and
the potential for adverse human health effects in these exposed populations.
In accordance with the responsibilities under the Great Lakes Critical Programs
Act of 1990, the USEPA transferred funds to the Agency for Toxic Substances
and Disease Registry (ATSDR), which has developed a research program to
address human health effects from exposure to contaminants in the Great Lakes.
Much of the historical data used in this report was obtained from information and
prior research performed by state health departments and universities. Federal
agencies other than the USEPA and ATSDR, such as the Centers for Disease
Control and Prevention (CDC), National Institute for Occupational Safety and
Health, and state agencies have all participated in preliminary reviews of this
report. This report reflects comments made by those agencies as well as those
made by the principal researchers whose work is contained within the report.
U.S. Environmental Protect.on .Reportto Con8ress
Region 5, Library (PL-12J)
77 West Jackson Boulevard, 12th Floor
Chicago, II 60604-3590
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FOREWORD (Contd.)
Because this research program is still in its early phases, there are no results of
these new studies presented in this report. However, preliminary results support
earlier reports of an association between the consumption of contaminated Great
Lakes fish and body burdens of persistent toxic substances, including PCBs,
organochlorines, and heavy metals such as mercury and lead. These body
burdens, identified in the fluids and tissues of consumers of Great Lakes fish
appear to be higher than those in the general population. Research studies are
ongoing to assess any effects of these contaminants on human health. The health
studies are investigating neurobehavioral and developmental deficits potentially
associated with the consumption of contaminated fish. In the meantime, the
potential human health effects in at-risk populations associated with documented
exposures and body burdens remain a potential public health concern. In those
instances of documented exposures and associated body burdens, the research
community supported by this program is pursuing appropriate public health
interventions and communication efforts for defined at-risk populations in the
basin with an emphasis on disease prevention. Results from all studies will be
summarized in a supplement to this document, upon completion of the studies.
Report to Congress
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ACKNOWLEDGMENT
This report was made possible by a number of dedicated people from
the Agency for Toxic Substances and Disease Registry (ATSDR), all
the investigators participating in the Great Lakes Human Health Effects
Research Program, and the USEPA Great Lakes National Program
Office (GLNPO). We extend a special thanks to ATSDR staff, Dr.
Barry Johnson, Assistant Administrator, Dr. Andrea Wargo, Associate
Administrator, Dr. Christopher DeRosa, Director of the Toxicology
Division, Dr. William Cibulas, Chief of Research Implementation
Branch, Dr. Hemline Hicks, Senior Environmental Health Scientist and
Program Manager, and Zemoria Rosemond, Environmental Health
Scientist. We would also like to thank the USEPA/GLNPO staff, David
Cowgill, Chief of the Remedial Programs staff, Darlene Punches,
Computer Specialist, Cynthia Bowman, Computer Assistant, Steve
Garbaciak, Environmental Engineer and Diane Dennis-Flagler, Envi-
ronmental Scientist for their help in assembling the final product.
Although there are certainly more people involved in the creation of this
report than can be named in this acknowledgement, the absence of those
names does not mean they are forgotton; we appreciate everyone
connected with this Program.
Hi Report to Congress
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EXECUTIVE SUMMARY
HEALTHY
PEOPLE 2000
In 1990, Congress passed the Great Lakes Critical Programs Act, amending the
Federal Water Pollution Control Act. This amendment requires the U.S.
Environmental Protection Agency (USEPA), in consultation with the Agency
for Toxic Substances and Disease Registry (ATSDR) and the Great Lakes states,
to submit to Congress a report assessing the adverse effects of water pollutants
in the Great Lakes basin on the health of persons in the Great Lakes states and
the health offish, shellfish, and wildlife in the Great Lakes system. In support
of this directive, Congress authorized funds not to exceed $3 million to carry out
research to support this report for each of the fiscal years 1992,1993, and 1994.
The USEPA received earmarked appropriations of $2 million for fiscal year
1992 and $3 million per year for fiscal years 1993 and 1994 to support human
health effects studies in the Great Lakes basin. These funds were transferred by
the USEPA's Health Effects Research Laboratory to the ATSDR to conduct
these studies. This report focuses on adverse human health effects as opposed
to fish, shellfish, and wildlife for two reasons. First, this was done because there
are few if any comprehensive reports addressing human health in the Great
Lakes. Conversely, there are a number of reports addressing the effects of Great
Lakes pollutants on fish and wildlife including the USEPA's 1994 Report to
Congress on the Great Lakes Ecosystem, (pages 11 to 25) and the U.S.Fish and
Wildlife Service (USFWS) Annual Report to Congress required by the Great
Lakes Fish and Wildlife Restoration Act of 1990. Second, from discussions
between USEPA and Congressional staff it was apparent that the Congressional
intent of this report was to address human health impacts. The Congressional
intent was also for the USEPA to take responsiblity for the report preparation
and transferring funds to the ATSDR. ATSDR's responsibilty is to oversee the
technical human health research program.
Although this report presents no results, it summarizes the existing literature on
Great Lakes human health studies related to contaminants, as well as research
progress made by the ATSDR's Great Lakes Human Health Effects Program
during fiscal year 1992. Final results and conclusions are expected to be
submitted as a supplement to this document upon completion of the studies.
ATSDR's mission is to prevent exposure and adverse human health effects and
diminished quality of life associated with exposure to hazardous substances from
waste sites, unplanned releases, and other sources of pollution present in the
environment. The activities described in this report support this mission and are
consistent with achieving the health promotion and disease prevention objectives
of Healthy People 2000, a national strategy put forth by the U.S. Department
of Health and Human Services for significantly improving the health of the
Report to Congress
IV
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EXECUTIVE SUMMARY (Contd.)
Nation during the next decade.
ATSDR made significant progress in implementing its Great Lakes Human
Health Effects Research Program in fiscal year 1992. The ATSDR research
program is designed to investigate and characterize the association between the
consumption of contaminated Great Lakes fish and short- and long-term harmful
health effects. The objectives of the research program are to:
• build upon and extend the results from past and ongoing research,
• develop information databases and/or research methodology that will
provide long-term benefits to the human health effects research efforts
in the Great Lakes basin,
• provide direction for future health effects research,
• provide health information to state/local health officials, the concerned
public, and their medical health care professionals,
• in concert with state and local health officials, increase public awareness
regarding the potential health implications of toxic pollution in the
Great Lakes basin, and
• coordinate as necessary with relevant U.S. Public Health Service (PHS)
research programs and activities, including those of the Food and Drug
Administration (FDA), Centers for Disease Control and Prevention
(CDC), National Institutes of Health (NIH), and the Indian Health
Service (IHS), as well as the U.S. Environmental Protection Agency,
and state and local health departments to ameliorate adverse public
health impacts of persistent toxic substances in the Great Lakes basin.
Toward this end, ATSDR developed a Great Lakes Health Effects Research
Strategy. The goal of this strategy is to identify human populations residing in
the Great Lakes basin that may be at greater risk of exposure to chemical
contaminants present in one or more of the Great Lakes and to help prevent any
adverse health effects. This strategy is built upon the five traditional elements
of disease prevention: identification, evaluation, control, dissemination, and
infrastructure. This strategy has been endorsed by the Council of Great Lakes
Research Managers and has been endorsed by the International Joint Commis-
sion as a framework for the study of human health and other ecosystem effects
in the Great Lakes basin.
For fiscal year 1992, the USEPA through ATSDR funded nine research grants
to study the potential adverse human health effects from consumption of
contaminated Great Lakes fish. These studies include eight epidemiologic
Report to Congress
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EXECUTIVE SUMMARY (Contd.)
investigations in presumed susceptible populations, (i.e., Native Americans,
sport anglers, the urban poor, pregnant females, and fetuses and nursing infants
of mothers who consume contaminated Great Lakes fish). The ninth study
focuses on developing more sensitive methods to detect persistent and
bioaccumulative Great Lakes contaminants such as polychlorinated biphenyls,
dioxins, alkylated lead, mirex, and methylmercury in human biologic tissues and
fluids. The 10 grants funded by ATSDR for fiscal year 1993 included nine
continuation awards for the studies initiated in 1992 and one new award that
established an interlaboratory-based quality assurance/quality control program
for the ATSDR research program. The fiscal year 1994 funds were used to
continue funding the previously mentioned ten projects.
The impact of this research program will be felt most directly by the communities
within the Great Lakes basin. Collectively, these 10 research projects will
• build upon and extend six existing human health studies in the Great
Lakes basin that include higher risk populations,
• establish two new subpopulations that include African American wo-
men and reproductive-age men and women between 18 and 34 years
of age,
• improve analytical methodology for detecting low levels of Great Lakes
contaminants in human biologic tissues and fluids, and in environmental
media,
• characterize exposure to all 11 critical Great Lakes contaminants
identified by the International Joint Commission, as well as to other
pollutants,
• determine profiles and levels (body burden) of Great Lakes contaminants
in higher risk populations,
• identify sensitive human health end points from exposure to Great
Lakes pollutants, (i.e., behavioral, developmental, reproductive, neuro-
logic, endocrinologic, and immunologic),
• investigate paternal and maternal exposure to Great Lakes pollutants
and assess the potential for resulting health effects in their children
(transgenerational effects),
• increase collaboration, cooperation, and communication among the
researchers in the Great Lakes basin, and
• provide public health information on potential human health effects
from exposure to Great Lakes pollutants through fish consumption to
the study populations, health care providers, and State and local health
departments.
Report to Congress vi
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EXECUTIVE SUMMARY (Contd.)
Additionally, the research conducted by this program will help delineate the
relationships among contaminant levels in the environment, exposure pathways,
tissue levels, and potential human health effects; allow for evaluation and
interpretation of data across all human health studies to facilitate a basin-wide
analysis on the pollution problem in the Great Lakes; and provide a model for
other ecosystem-level studies intended to determine human health impacts of
hazardous waste.
vii Report to Congress
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Table of Contents
Foreword. j
Executive Summary /v
I. Introduction
Historical Overview ]
Persistent Toxic Substances 2
Pathways of Exposure 3
II. A Review of the Great Lakes Human Health Literature
Michigan Sport Fishermen Study 4
Wisconsin Sport Fish Consumers Study 5
Minnesota Ecologic Epidemiologic Study 6
Michigan Maternal and Infant Study 6
Smith's Wisconsin Maternal and Infant Study 9
Dar's Wisconsin Maternal and Infant Study 10
New York Ecologic Epidemiologic Study 10
III. Human Health Studies Outside of the Great Lakes -
Exposure to Similar Persistent Toxic Substances 11
IV. Limitations of Human Health Studies 13
V. Program Areas of Research
Characterization of Exposure and Determination of the
Profiles and Levels in Human Biologic Tissues and Fluids 17
Identification of Sensitive and Specific
Human Reproductive End Points 18
Determination of the Short- and Long-Term Risk(s)
of Adverse Health Effects in the Children of Exposed Parents 18
Establishment of Registries and/or Surveillance
Cohorts in the Great Lakes 19
VI. ATSDR Great Lakes Human Health Effects Research Program
Report to Congress viii
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Table of Contents (Cont'd.)
Great Lakes Health Effects Research Strategy 22
Research Program Objectives 24
Program Areas of Research 26
Impact ofATSDR Research Program 28
Research Program Timeline 31
VII. ATSDR Great Lakes Research Projects
Synopsis of Funded Awards for FY1992 32
PCB and DDE Exposure among Native American
Men from Contaminated Great Lakes Fish and
Wildlife 32
An Assessment of a Human Population at Risk: The
Impact of Consuming Contaminated Great Lakes Fish
on Native American Communities 38
Cognitive and Motor Effects of PCB Exposure in
Older People from the Michigan Fisheater Cohort:
Emphasis on the Role of Ortho-Substituted Congeners 42
The New York State Angler Cohort Study: Exposure
Characterization and Reproductive and
Developmental Effects 49
Great Lakes Fish as a Source of Maternal and
Fetal Exposure to Chlorinated Hydrocarbons 54
Contribution of Nursing to Behavioral Changes in
Children of Mothers Who Consumed Lake Ontario
Fish: Two Methodological Approaches 60
Health Risks from Consumption of Great Lakes Fish 65
Consortium for the Health Assessment of Great Lakes
Sport Fish Consumption 72
Congener-Specific PCB Analysis for Improved
Toxicity Assessment 83
Michigan Great Lakes Health Studies 87
VIII. References 90
ix Report to Congress
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Section I
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I. INTRODUCTION
Historical Overview
The Great Lakes hold one-fifth of the fresh water on the earth's surface, and
it is a valuable resource for both the United States and Canada. Approximately
25 percent of the Canadian population and 10 percent of the United States
population live within the Great Lakes watershed. For more than 200 years,
the Great Lakes basin has been used as a resource for industry, agriculture,
shipping and recreation. By the early 1960's, the environmental quality of the
Great Lakes had deteriorated significantly as a result of eutrophication,
overfishing, and the widespread presence of toxic substances. During the
1970's, the use of the Great Lakes as a disposal site for agricultural, industrial
and domestic wastes became an increasingly widespread concern due to
detrimental effects on fish and wildlife, and the potentially adverse effects on
human health.
Some pollutants of the Great Lakes have been found in significant concentra-
tions in fish and wildlife (Keith 1966; Aulerich and Ringer 1977; Nisbet 1989).
Wildlife investigators have demonstrated that these pollutants are implicated
in interfering with reproduction in birds and lake trout, contribute to congenital
abnormalities in birds, and have implicated these pollutants in the development
of neoplastic tumors in bottom-dwelling fish (Gilbertson et al., 1976; Maccubbin
etal., 1985; Price and Weseloh 1986). Concentrations of PCBs in some Great
Lakes fish have been significant enough at least since the mid-1970's to
warrant advisories to restrict consumption of these fish, particularly by
children and women of childbearing age.
Except for some migratory waterfowl, the fish and wildlife in the Great Lakes
basin generally live their entire lives within the basin, being continually
exposed to toxic contaminants in the water and/or food. A combination of
factors contribute to the Great Lakes acting as a reservoir for these relatively
nondegradable lipophilic chemicals. Factors such as the following: 1) long
retention times, the time it takes for the lake water to be completely exchanged
ranges from about 3 years for Lake Erie to 173 years for Lake Superior; 2)
internal recycling of contaminants; pollutants tend to adsorb onto particles and
eventually settle to the bottom of the lakes. However, the surficial sediments
are subject to resuspension by storm actions and other natural processes, and
the contaminants are dispersed throughout the water column again.
The concentrations of contaminants in the water column of the Great Lakes
declined measurably after significant decreases in the loadings of the pollut-
ants. Sediment-water interactions, however, greatly extend the overall re-
sponse time of the system to loading reductions. For example, data from mass
balance studies and inventories in the Great Lakes system indicate that there
is a reservoir of PCBs in the sediments that will continue to release PCBs into
n>
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the lakes at significant rates for decades to come (Andre et al., 1993; Beltran,
1992; Richardson, 1993; U.S. EPA 1989).
Fish tissue concentrations of some contaminants, including DDT and PCBs,
have declined significantly in the Great Lakes basin since the mid-1970s. At
that time, PCB concentrations in lake trout from Lake Michigan were about
22mg/kg. Bytheearly 1980'sthey had declined to about 6 mg/kg, a level not
greatly more than found now. Recently, the rate of decline of PCB concentra-
tions in these fish has slowed, and smaller incremental reductions in the fish
tissue levels will continue to occur.
As stated previously, fish tissue contaminant levels are lower than in the
1970's, which is when some of the early human health studies were done.
However more recent human studies that have taken advantage of newer
robust tools (e.g., improvements in analytical capabilities and clinical testing
sophistication) continue to demonstrate the potential for subtle effects in
exposed at risk human populations.
In 1972, the United States and Canada signed the Great Lakes Water Quality
Agreement to initiate remedial actions to improve the quality of Great Lakes
waters, focusing on the issue of excessive loadings of nutrients. In 1978, the
two countries signed a revised Agreement to "restore and maintain the
chemical, physical and biological integrity of the waters of the Great Lakes
ecosystems" (International Joint Commission 1978), with an emphasis on
reducing toxic contaminants. The Agreement was amended again in 1987
expanding many of the programs and recognizing that an ecosystem approach
was required to truly restore and protect the Great Lakes ecosystems.
Persistent Toxic Substances
The presence of toxic substances in the Great Lakes continues to be a
significant concern in the 1990s. In the U.S., some 70,000 commercial and
industrial compounds are now in use. More than 30,000 chemicals are
produced or used in the Great Lakes region. There have been 362 contami-
nants identified in the Great Lakes system; of these 362 approximately one-
third have been evaluated for their potential toxic effects on aquatic life,
wildlife, and human health. (An Inventory of Chemical Substances Identified
in the Great Lakes Ecosystem ,Vol. 1-6. IJC, Windsor, Ontario 1983 and
Federal Register March 23, 1995 Page 15367) in 1985, 11 of the most
persistent and widespread toxic substances were identified as "critical Great
Lakes pollutants" by the International Joint Commission (IJC). The critical
pollutants identified by the IJC are:
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CRITICAL POLLUTANTS
polychlorinated biphenyls (PCBs)
dichlorodiphenyl trichloroethane (DDT) and metabolites
benzo[a]pyrene (a member of a class of substances
known as PAHs)
hexachlorobenzene
alkylated lead
methylmercury
toxaphene
mirex
furans
dieldrin
dioxins
All 11 of the persistent toxic substances tend to bioaccumulate in organisms,
biomagnify in food webs, and persist at high levels in some areas of the
ecosystem of the Great Lakes. In the Great Lakes basin, 43 geographical
locations in the United States and Canada have been identified as "Areas of
Concern" because of high concentrations of these toxic pollutants. Of these
43 locations, 26 are located within the boundaries of the United States and 5
are shared with Canada, (i.e., connecting channels.
Eight of these pollutants are organochlorine compounds and are potentially
harmful because of their persistence, ubiquity, and associated toxicity. Both
alkylated lead and methylmercury are heavy metals that are potentially
harmful because of their chemical characteristics and demonstrated toxicity.
Eight of the 11IJC critical pollutants are lipophilic and biomagnify within the
aquatic food chain, thereby making them available to higher forms of life,
including humans. PAHs, methylmercury, and alkylated lead also
bioaccumulate in human tissues.
Pathways of Exposure
Potential environmental pathways of human exposure to Great Lakes pollut-
ants include inhalation of air, ingestion of water, foodstuffs or contaminated
soil, and dermal contact with water or airborne particulates. Multimedia
analyses indicate that the majority (80-90%) of human exposure to chlori-
nated organic compounds comes from the food pathway, a lesser amount (5-
10%) from air, and minute amounts (less than 1%) from water (Birmingham
et al. 1989; Newhook 1988).
Most of the data available on human exposure to toxic substances in the Great
Lakes come from the analyses of contaminant levels in drinking water and
sport fish. The consumption of contaminated sport fish and wildlife can
significantly increase human exposure to the Great Lakes critical pollutants.
A spectrum of these major contaminants has been identified in cooked Great
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Lakes fish. Investigators have demonstrated that blood serum levels of these
contaminants are significantly increased in consumers of contaminated Great
Lakes sport fish as compared to nonfisheaters (Humphrey 1983a,b; Jacobson
et al. 1989).
Even though residents of the Great Lakes basin are exposed to toxic
substances from many sources originating within and outside the region, the
main routes of human exposure to contaminants from the waters of the Great
Lakes are ingestion of fish and to a lesser extent ingestion of drinking water
(National Health and Welfare Canada 1991). Also, several investigators have
shown that exposure from fish far outweighs atmospheric, terrestrial, or water
column sources (Swain 1983; Humphrey 1983b). These patterns may vary
for populations living in the vicinity of industrialized areas such as refineries
or smelting plants.
Several epidemiologic investigations have been conducted to investigate the
association between water pollutants in the Great Lakes and the health of
people in the Great Lakes states. These studies have demonstrated increased
tissue levels of toxic substances (body burdens) in these populations that may
be associated with or potentially result in reproductive, developmental,
behavioral, neurologic, endocrinologic, and immunologic effects. The
following review of human health literature in the Great Lakes basin
summarizes and compares the findings concerning exposure to Great Lakes
pollutants.
II. A REVIEW OF THE GREAT LAKES HUMAN HEALTH
LITERATURE
Several human health studies have been conducted in the Great Lakes basin.
The following review summarizes some of these studies and other studies
concerning exposure to Great Lakes pollutants.
Michigan Sports Fishermen Cohort Study
The Michigan Sports Fishermen Cohort study was initiated in 1974 to assess
the potential exposure of humans to PCBs from consumption of sport fish
from Lake Michigan (Humphrey 1976). The population consisted of 336
adults from 18 western counties of Michigan that border Lake Michigan.
These adults made up two groups: those who consumed large quantities of
Lake Michigan sport fish, especially trout and salmon, and those who did not
eat fish from Lake Michigan. Blood samples were taken from each group and
the serum was analyzed for PCB compounds. The resulting data were
compared to information received from each individual through interviews.
Fish consumption variables were compared with PCB levels. The results
from this initial study demonstrated a statistically significant correlation
between fish consumption patterns and PCB levels in this population.
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In the late 1970s and early 1980s, 123 members of this original cohort were
re-examined using the same protocol. On average, sport fish-eaters consumed
32 pounds of fish a year, and some ate as much as 262 pounds a year. This
average was approximately five times the national per capita fish consump-
tion rate commonly used in risk estimates at that time. Individuals who
regularly ate 24 pounds a year or more of Great Lakes fish, especially trout and
salmon, had higher (P < 0.001) serum concentration levels of PCBs than
individuals who seldom or never ate such fish.
A third cohort was included in these studies to test the above conclusion. In
1982, 572 individuals from the Lake Michigan shore who ate 24 or more
pounds of fish a year were compared with 419 randomly selected residents
from the same communities along the Lake Michigan shoreline. Again, the
data indicated that individuals in the upper range of fish consumption had
serum PCBs concentrations approximately four times greater than those of
unexposed individuals. These studies support a positive correlation between
human uptake of toxic pollutants in the Great Lakes and Great Lakes fish
consumption (Humphrey 1976, 1983a, 1988a, 1988b, 1989).
Wisconsin Sports Fish-Consumers Study
Another study of sport anglers conducted in Wisconsin (Wisconsin Sports
Fish-Consumers Cohort Study) was similar in design to the Michigan Sports
Fishermen Cohort Study. Investigators in Wisconsin surveyed 801 sport
anglers for their fishing and consumption habits, and for comprehension of
and compliance with the Wisconsin fish consumption health advisory (Fiore
etal. 1989; Sonzognietal. 1991). In this cross-sectional study of Wisconsin
anglers, the investigators also measured the angler's body burden levels of
PCBs and DDE to examine possible correlations between sport fish consump-
tion and the body burden of these substances. Of the 801 persons surveyed,
198 individuals provided blood samples for analysis of PCBs and DDE.
The survey results indicated that the mean number of sport fish meals eaten
annually was 18,72 percent of the anglers were aware of the fish advisory, and
57 percent changed their fishing or fishing habits as a result of the advisory.
Of the 198 blood samples drawn for PCBs and DDE analysis, 6 were
determined to be inadequate for analysis. The mean PCB serum level for the
192 anglers was 2.2 ug/1; of the 13 PCB congeners for which standards were
available, only 2 were not detected in sera. The investigators also determined
that the congeners most frequently identified in human sera were also the most
abundant congeners in the tissues of a variety of Wisconsin fish (Maack and
Sonzogni 1988). Additionally, there was a statistically significant Spearman
correlation (R = .21, p = .003) between the PCB congener sum and the total
number of sport fish meals. The mean DDE level was 6.3 ug/1. Of the 109
samples tested, 65 of the subjects had a serum level between 5.0 and 10.0
ug/1, and 44 had levels from 10.0 to 40.0 ug/1. There was also a statistically
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significant positive correlation (R =0.14, p =0.04) between sera levels and
the total number of sport fish meals.
The Wisconsin Sports Fish-Consumers Cohort is the first study of anglers for
whom PCB-specific congeners were determined in human sera. Using
capillary column gas chromatography as opposed to the older, packed column
technique, these researchers were able to detect PCB congeners 153,138,180,
and 118 (order based on frequency of occurrence). The authors also suggest
that the capillary method produces PCB results that are approximately 30
percent more sensitive than those obtained by packed column chromatogra-
phy, allowing for the summation of individual congeners to estimate total
PCB burden. The differences in laboratory techniques between the Wisconsin
and Michigan studies (used packed column chromatography) makes the
comparison of results very difficult.
Minnesota Ecologic Epidemiologic Study
In Minnesota, a researcher conducted an eco logic epidemiologic study to
examine fetal and neonatal death rates, as well as congenital malformations
rates of residents in counties that were in close proximity to the lakes
(Schuman et al. 1982). An ecologic study is a descriptive study in which
measures that represent characteristics of entire populations are used to
describe disease in relation to some factors of interest. In ecologic studies the
unit of observation is a group of people rather than individuals and the unit of
exposure pertains to geographical areas rather than to individuals. The
assumption in this study was that residents in lake- bordering counties were
more likely to have environmental and occupational exposure to pollutants
discharged into the air and water. Information on the above human end points
was collected for 1950-1975 and 1977 for six of the eight Great Lakes States
(excluding New York and Pennsylvania) and analyzed. The investigator
found no significant differences among fetal death rates, neonatal death rates,
and congenital malformations in populations bordering the lakes; however,
results are considered inconclusive because the rates were not adjusted for
confounding variables, such maternal age, race, or sex.
Michigan Maternal and Infant Study
In Michigan, the Maternal/Infant Cohort Study was designed to assess the
impacts of consumption of contaminated fish on pregnant women and their
newborn infants. The Michigan Maternal/Infant Cohort Study consisted of
313 infants and their mothers from four hospitals in the Michigan area (Fein
et al. 1983). The 313 infants included 242 infants born to women who
consumed moderate to high amounts of Lake Michigan fish for at least 6 years
before their pregnancy (average consumption was 6.7 kilograms/year, which
is equivalent to about 2'/z pounds of salmon or lake trout meals a month), and
71 infants whose mothers did not eat Lake Michigan fish.
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The initial study examined two parameters: (1) the relationship between
maternal fish consumption and exposure to PCBs and (2) the potential for in
utero PCB exposure based on maternal fish consumption. The investigators
analyzed for PCBs in maternal serum and in umbilical cord blood. The
authors reported that no relationship was found between PCB levels in
umbilical cord serum and maternal fish consumption. However, a statistical
correlation was found between the maximum yearly ingestion of Lake
Michigan fish and PCB levels in maternal serum. The concentration of PCBs
in serum was predictable from the quantity of fish eaten by the mothers.
These studies also indicated a highly significant (P< 0.0001) correlation of
umbilical cord serum to maternal serum, which strongly suggests transpla-
cental transfer of PCB molecules (Jacobson et al. 1983, 1984a; Schwartz et
al. 1983).
Several adverse developmental health outcomes were observed in newborn
infants of mothers who consumed more than 11.8 kilograms of contaminated
fish (Fein et al. 1984). These health outcomes were birth weight, head
circumference, and gestational age, as estimated by the mothers' reports of
their last menstrual periods and on the Ballard Examination for Fetal
Maturity. The exposed infants showed statistically significant decreases in
infant birth weights (160 to 190 grams lighter than controls) and gestational
age (average 4.9 days less than controls), and head circumference (average
0.6 centimeters smaller than controls). The authors attributed these effects to
intrauterine exposure to PCBs via the mothers' diet of contaminated Lake
Michigan fish. Maternal serum PCB concentrations for the samples as a
whole averaged 5.5 ± 3.7 ng/ml. According to the authors, this value is
comparable to other midwestern American samples. Reported maternal fish
consumption did predict maternal serum PCB levels (r= 0.37, P < 0.001),
which in turn predicted umbilical cord PCB levels in their infants (r= 0.41,
P < 0.001). The infants' umbilical cord serum averaged 2.5 ±1.9 ng/ml.
A follow up of these infants at 5 months postpartum indicated that the
observed difference in birth weights still persisted (Jacobson and Jacobson
1988). Neonates who were exposed to 3 ppb or greater of PCBs, as indicated
in their umbilical cord serum, were approximately 260 grams lighter than
those infants exposed to less than 3 ppb PCBs in their cord serum. Further
studies of these children at 4 years of age indicated a continuous deficit in
weight gain when compared to the controls. Children with cord serum PCB
levels of 5.0 ng/ml or more weighed 1.8 kg less on average than the lowest
exposed children after adjustment for covariates and potential confounders
(Jacobson et al. 1990a,b).
Neurological effects were also observed in the infants of this Michigan study.
Prenatal exposure to low levels of PCBs from maternal consumption of
contaminated Great Lakes fish was shown to have result in subtle behavioral
effects (Jacobson et al. 1984a,b,c).
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The Brazelton Neonatal Behavioral Assessment Scale (NBAS; Brazelton
1973) was used to assess behavioral outcomes, and the Ballard Examination
for Fetal Maturity (Ballard et al. 1979) was administered to assess neuromus-
cular and physical maturity in 242 infants from the Michigan Maternal/Infant
Cohort, 48 to 72 hours following birth (Jacobson et al. 1984b,c). Neonatal
behavioral deficits associated with maternal consumption of contaminated
fish included greater inclination to startle; poorer motor reflex and neuromus-
cular functioning; and depressed responsiveness, as evidenced by a greater
number of hypoactive reflexes and limited adaptability of states of mind when
compared to control infants. The degree of maternal fish consumption
predicted a linear combination of alterations in neonatal health, including
autonomic maturity (p >.025), number of abnormal reflexes (p >.05), and
range of state (p >.005). Such subtle effects are categorized in the "worrisome"
range oy Als et al. (1979).
In the same infants from the Michigan Maternal/Infant Cohort, umbilical cord
serum samples were analyzed for the presence of PCBs (Aroclors 1016 and
1260) as a direct measure of infant exposure. However, the authors found no
correlation between cord serum values and behavioral effects in infants
(Jacobson et al. 1984a). Difficulty in obtaining cord serumlevels for all of the
infants, especially those with the highest fish consumption exposure levels
may have biased these results. Also, the behavioral effects associated with fish
consumption in the Michigan Maternal/Infant Cohort may be due to unmeasured
contaminants, other than PCBs, present in the fish (Jacobson and Jacobson
1988; Stone 1992).
When the infants from the Michigan Maternal/Infant Cohort reached 7 months
of age, Jacobson et al. (1985) showed that infant visual recognition memory
was predicted by both cord serum PCB levels and maternal report of Lake
Michigan fish consumption, but not postpartum PCB exposure from breast-
feeding (Jacobson and Jacobson 1988). Visual recognition memory perfor-
mance was assessed using Pagan's Visual Recognition Memory Test (Pagan
and McGrath, 1981) in infants at 7 months. Although the infants used in this
study appeared clinically normal at birth, poorer performance on Pagan's Test
was associated with intrauterine PCB exposure (measured by umbilical cord
serum PCB levels and maternal fish consumption) indicating that some
behavioral deficits resulting from secondary exposure may not be evident at
birth and that follow up examinations are necessary. Jacobson and Jacobson
(1988) concluded that visual recognition memory testing is capable of
detecting subtle performance deficits in otherwise clinically normal infants.
Reduced performance on recognition memory testing in children exposed to
contaminants through maternal consumption of Great Lakes fish may result
from processing of information at a slower speed, poor visual discrimination,
poorer attention, or difficulties in encoding and retrieving of information
(Jacobson and Jacobson 1988). In addition, infant recognition memory has
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been shown to be a valid predictor of later intelligence, as measured by verbal
intelligence quotient (IQ) scores among children (Pagan and McGrath 1981).
Therefore, it is plausible that poorer performance in memory testing at an early
age may be indicative of long-term cognitive deficits. Animal data subjected
to the risk assessment model proposed by the U.S. Environmental Protection
Agency (EPA) for developmental toxicity (IRIS 1994) resulted in derivation
of a "reference dose" for PCBs of 7 X 105 mg/kg/day, based on reduced birth
weights. The ATSDR has derived a chronic oral Minimal Risk Level (MRL)
for PCBs of 0.00002 mg/kg/day, based on immunological effects (ATSDR
1993).
Additional follow up studies evaluated 4-year-old children from the Michigan
Maternal/Infant Cohort previously examined for PCB-related deficits as
infants (Jacobson et al. 1989, 1990a,b). At age 4, study participants were
administered the McCarthy Scales of Children's Abilities (McCarthy 1972) to
assess cognitive function. Exposure due to maternal consumption of contami-
nated fish from Lake Michigan was measured by PCB levels in cord serum
(average = 2.5 ± 2.0 ng/ml), maternal serum (average = 5.9 ± 3.6 ng/ml),
maternal milk (average = 835.9 ± 388.4 ng/ml), and body burden at 4 years of
age (breast-fed for at least 6 months, 5.1 ± 3.9 ng/ml; breast-fed for less than
6 months, 1.2 ± 1.6 ng/ml; and 0.3 ±0.7 ng/ml for those not breast-fed). High
cord serum PCB levels predicted poorer performance on McCarthy Verbal and
Memory Scales in a dose-dependent manner. These results are consistent with
earlier findings of an association between poorer memory and prenatal
exposure in infants (Jacobson et al. 1985).
Postnatal exposure from nursing based on maternal milk PCB levels and
weeks of nursing was not related to memory deficit in these children (Jacobson
et al. 1989). In addition, the children's serum PCB level at 4 years (related
primarily to postnatal nursing) also failed to predict cognitive outcomes in this
study. Although a larger quantity of PCBs is transferred postnatally through
lactation, prenatal exposure was found to be associated with decreased
cognitive function in young children. In summary, findings of the Fein et al.
(1983, 1984) and Jacobson studies are significant in that both initially
identified associations between maternal consumption of contaminated fish
and adverse human health outcomes in their children.
Smith's Wisconsin Maternal and Infant Study
The Wisconsin Maternal and Infant Cohort study also investigated the
association of maternal exposure to toxic substances and adverse health effects
in infants (Smith 1984). Results from this study indicated an association
between maternal serum PCB levels and the number and type of infectious
diseases suffered by their infants. The categories of infectious diseases
included colds, earache, and flu symptoms. The author concluded that in utero
exposure to PCBs resulted in an increase in the number of illnesses in infants
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during the first 4 months of life. However, the design of this study prevented
access to either umbilical cord blood or infant serum acquired immediately after
birth to analyze for the presence of PCB s. Therefore, the issue of exposure was
presumed and not actually addressed by the Wisconsin study (Swain 1991).
Par's Wisconsin Maternal and Infant Study
A more recent study in Wisconsin examined Great Lakes fish consumption and
reproductive outcomes in Green Bay (Dar et al. 1992). In this prospective study
1,112 women who were seen at the time of a positive pregnancy test were asked
to complete a questionnaire on fish consumption, health and reproductive
history, and other relevant issues, and to provide blood samples for PCB
analysis. After the pregnancy, the following reproductive outcomes were
abstracted from hospital labor reports and measured: fetal wastage and still-
births, birth weight, birth length, head circumference, ponderal index, and birth
weight percentiles for live births. The typical negative associations between
birth size measures and confounding variables, (i.e., consumption of caffeine
and alcohol, and smoking) were found. Birth size was positively associated
with gestational age, birth order, weight gain during pregnancy, male babies,
and rural residence. However, contrary to expectations, a positive association
was found (p<0.044) between birth size and PCB exposure (via sport fish
consumption) for most mothers (the exception being those mothers who gained
more than 34 pounds during pregnancy).
Serum PCB levels were performed on 106 women in this group, and a positive
correlation (Pearson p = 0.666) was found between the amount of Lake
Michigan fish mothers reported to consume and their PCB serum levels (Dar
et al. 1992). The PCBs concentration was based on the sum of 13 individual
congeners; only 5 of 13 PCB congeners were identified in maternal serum.
Also, only 23 percent of the study group had PCBs above the detection limit of
0.6 ng/ml for each congener. Additionally, the consumption of highly
contaminated sport fish was very low in this cohort. The authors concluded that
the Green Bay mothers were exposed to very low levels of PCBs, compared
with exposures in other studies,(e.g. Fein et al. 1983,1984). This low estimated
level of exposure does not appear to have had a negative effect on birth size. The
authors also suggest that if adverse reproductive effects occur in infants because
of maternal fish consumption, perhaps there is a threshold effect below which
there are no negative effects.
New York Ecologic Epidemiologic Study
An ecologic epidemiologic study was conducted within the Great Lakes
drainage basin of upstate New York (Kagey and Stark 1992). The hypothesis
for this study was "maternal residence within the Great Lakes drainage basin
from 1968-1987 adversely impacted birth weight, gestational age, and congeni-
tal anomaly rates of their offspring. In addition, within the basin area, those
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infants whose mothers resided within the counties adjacent to the lakes were at
greater risk of lower birth weight, shorter gestational age, and increased
congenital anomaly rates associated with their close proximity to the lakes." In
defining the exposure area as the Great Lakes drainage basin, it was assumed
that potential exposure to contaminants was through one or more of the
following routes: inhalation of specific contaminants that evaporated from the
lakes, consumption of sport fish, home garden produce, and drinking water
from the lakes or from point sources within the basin.
The authors determined that the mean birth weight of infants born within the
Great Lakes drainage basin was comparable to the mean birth weight of infants
born outside the basin: 3,313 versus 3,319 grams, respectively. Mothers
residing in counties adjacent to the Great Lakes gave birth to infants who
weighed 28 grams less than infants born to mothers who resided in nonadjacent
areas. Approximately 6 percent of all births in this study population were low
birth weight (less than 2,500 grams). This percentage was slightly higher in
infants born to mothers who resided near the lakes, however.
The mean gestational age for the entire study population was 279 days; no
differences were found in mean gestational age by study area or density quintile.
Differences in congenital anomaly prevalence rates were also difficult to detect
because of changes in coding and reporting practices over time. The authors
concluded that the two major limitations of ecologic epidemiologic studies
were the lack of individual measurements on exposure or outcome and the
inability to control for confounding variables
III. HUMAN HEALTH STUDIES OUTSIDE OF THE GREAT
LAKES - EXPOSURE TO SIMILAR PERSISTENT TOXIC
SUBSTANCES
Other epidemiologic studies have been conducted on mothers exposed to toxic
substances, similar to those identified in Great Lakes fish, whose children have
exhibited either reproductive and developmental or neurobehavioral effects.
The North Carolina Breast Milk and Formula Project
This study investigated the effects of prenatal exposure to PCBs (Rogan et al.
1986). Mothers were exposed to environmental or background levels of PCBs.
A total of 856 breast-fed infants were identified at birth and periodically
examined for up to 60 months. No association was found between maternal
PCB levels and birth weight, head circumference, or gestational age. However,
neurological effects were observed in these children. Children with higher in
utero PCB exposure (assessed as breast milk fat PCBs concentration) exhibited
neurological deficiencies, such as hyporeflexia and hypotonicity similiar to
those observed in the Michigan Maternal and Infant Cohort Study.
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At 6 and 12-month follow up examinations, usingthe Bayley's Scales of Infant
Development, the psychomotor index indicated continued decreases in behav-
ioral effects (Gladen et al. 1988). At subsequent examinations these behavioral
effects were not observed (Gladen and Rogan 1991).
Japan and Taiwan PCBs Studies
Two separate but similar studies were conducted on accidental exposure to
PCBs involving human populations in Japan and Taiwan (Hsu et al. 1985). In
Japan in 1968 and in Taiwan in 1979, accidental mass poisoning incidents
occurred from cooking oil contaminated by thermally degraded polychlori-
nated biphenyls. In 1985,117 children born to women in Taiwan who ingested
contaminated rice oil and 108 unexposed controls were examined and evalu-
ated.
The authors indicated that the exposed children were shorter and weighed less
than the controls. Additionally, the exposed children showed delayed develop-
mental milestones, deficits on formal developmental testing, and abnormalities
on behavioral assessment. Intrauterine exposure of Japanese infants to PCBs
was associated with prematurity and decreased size for gestational age (Higuchi
1976; Wong and Huang 1981). In follow up studies of these exposed children,
growth deficiencies were evident. Autonomic disturbances were observed;
sluggish, clumsy, and jerky movements were exhibited; and the average
intelligence quotient in these children was in the low 70s. The children in this
study were exposed to PCBs but also to the heat degradation products of PCBs
including highly toxic poly chlorinated dibenzofurans.
Occupationally Exposed Female Capacitor Workers
This study investigated the relationship of PCB exposure to birth weight and
gestational age in 356 infants of mothers occupationally exposed to PCBs
during the manufacture of capacitors in upstate New York (Taylor et al. 1989).
The results from this study indicated that infants exposed in utero to PCBs were
also smaller at birth, on average 153 grams less than unexposed infants, and that
their mean gestational age was 6.6 days shorter than infants from mothers
employed in the same facility but not directly exposed to PCBs. The authors
concluded that there is a significant association (p<0.02) between increasing
serum PCB level and decreased birth weight and gestational age, and that the
decrease in birth weight is at least partially related to shortened gestational age.
The authors also stated that the magnitude of these effects was quite small and
the clinical significance of these results was questionable. However, the
authors suggest that unknown or unmeasured confounding variables that
influence birth weight and gestational age will of course alter the results of their
study.
Epidemiologic studies of exposed human populations usually provide the most
convincing evidence of human health effects. However, epidemiologic studies
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may vary with design (i.e., prospective studies, retrospective studies, ecologic
studies) or investigators may use different analytical techniques; therefore, it
becomes very difficult to evaluate or compare studies. The following section
on study limitations will discuss some of these differences.
IV. LIMITATIONS OF HUMAN HEALTH STUDIES
The Michigan Sport Fishermen Cohort Studies was the first epidemiologic
study to assess exposure to PCBs in humans from consumption of sport fish
from the Great Lakes. This study supports a positive correlation between
human uptake of toxic pollutants in the Great Lakes and Great Lakes fish
consumption. Generally, this study was well designed and executed but limited
by the analytical methodology used, which was state-of-the-art at the time. For •
example, to determine PCB levels in serum, the samples from the cohort were
pooled to determine "total PCBs." This methodology did not allow study of
individual PCB levels within a cohort. Additionally, the investigators used
packed column chromatography to determine total PCBs, which resulted in a
lack of sensitivity to detect individual PCB congeners. But, again the
methodology used in these late 1970s studies was the best available at the time.
The differences in laboratory techniques makes evaluating or comparing
studies very difficult. For example, the two sport angler studies (Michigan and
Wisconsin) were both conducted to determine the possible correlation between
sport fish consumption and body burden of toxic substances. The analytical
methods used to determine total PCBs in human sera were packed column in
the Michigan study and capillary column gas chromatography in the Wisconsin
study. The interlaboratory differences strongly contributed to variation in
levels of total PCB levels in sera in the two studies. The highest PCB congener
sum in the Wisconsin study was 27.1 ug/1; the highest value in the Michigan
study was 366 ug/1. Using the older packed column technique, the investigators
could only determine total PCB. Using capillary column gas chromatography
the investigators could determine PCB by summation of individual congeners
to estimate total PCB body burden. The Michigan and Wisconsin values may
be attributed to design differences, (e.g., different methods of selecting survey
participants such as age and sex), different methods to obtaining fish consump-
tion histories, different times of sample collection [ 1973 and 1980 for Michigan
versus 1986 for Wisconsin], or different consumption and fish preparation
patterns). Because of the differences in analytical methodology and design, the
results of these studies are therefore limited in providing convincing evidence
of the status of human health from exposure to Great Lakes contaminants.
The Michigan Maternal/Infant cohort study is widely referenced because it was
the first epidemiologic investigation that demonstrated the association between
maternal consumption of Great Lakes fish and adverse human health outcomes
in their children. The establishment of this cohort was an excellent idea in an
attempt to associate maternal fish consumption, PCB serum levels, and adverse
health outcomes. However, several shortcomings and unanswered questions
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regarding this study remain. The following discussion briefly evaluates the
major limitations of this study.
Of the 8,482 women delivering infants in four Michigan hospitals, only 4
percent (343) were considered eligible to participate based on reported high fish
consumption (Fein et al. 1983,1984). The control population comprised only
71 women who had reported eating no Great Lakes fish. The statistical power
of the study is limited by the size of the control group, which is only one-fifth
the size of the exposed group.
A nonrandom sampling technique was used to define the study population.
Investigators purposely selected individuals who reported consuming large
amounts of Lake Michigan fish; therefore, the final study group consisted of a
small homogeneous population. Random sampling and heterogeneity of the
investigative cohort are important because the ideal would be to have the
unbiased ascertainment of eligible exposed and control individuals.
A number of differences existed in individual characteristics within the exposed
group, as well as between the exposed mothers and the control group in these
studies. Thirty-seven potentially confounding factors that may have affected
the observed health outcomes in these studies have been identified (Fein et al.
1984). These factors include health history, maternal health, obstetrical care,
pregnancy and delivery, and fetal exposure to drugs, such as alcohol, caffeine,
nicotine, and cold medications. These differences raise the possibility that the
exposed and control groups may differ in their overall health condition and that
these differences may affect their pregnancy outcome or infant development or
both.
Estimation of maternal fish consumption in this study was based on the ability
of the mother to recall past patterns offish consumption for at least 6 years. The
investigators defined the total fish consumption using a "weighted sum"
(amount of fish meals consumed a year) calculation. The accuracy of this
method is questionable, especially when trying to determine maternal expo-
sure, and precluded the investigators from making dose comparisons.
In addition, maternal estimates of cumulative fish consumption is poorly
correlated with maternal serum PCB levels in some studies. In other studies,
there are correlations between fish consumption and serum measurements. The
reason for these discrepancies is unknown. Also, there was no correlation
between maternal fish consumption and umbilical cord levels of PCB which are
considered to be an accurate measure of transplacental exposure. Given these
limitations, it appears that either the estimation of maternal fish consumption
was inaccurate or the measurement of total PCB levels was not a good indictor
of total fish consumption. Also, the standards used (Aroclor 1016 and 1260)
as references to quantify total PCBs accounted for only a small portion of the
PCB congeners detected (Swain 1991). Therefore, the analytical methods used
to measure PCB levels may not have been appropriate.
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In summary, the limitations of this study were the sampling technique used to
define the population, uncertainties associated with estimation of exposure,
lack of correlation between fish consumption and PCB levels in serum in some
studies, and a number of confounding variables not taken into account. In
addition, the health effects observed in this study may also be attributed to
exposure to contaminants other than PCBs present in fish. Researchers have
demonstrated that the fish consumed probably contained a mixture of other
chlorinated toxic chemicals, such as DDE, DDT, aldrin, and dieldrin, as well
as heavy metals, such as mercury or lead. These substances have been found in
contaminated fish from the Great Lakes region (National Health and Welfare
Canada 1991). Currently, follow up studies using the Michigan Maternal/
Infant Cohort are under way. New information from these efforts may provide
answers for some of the remaining questions.
In Dar's Wisconsin Maternal and Infant study which examined fish consump-
tion and reproductive outcomes in women from Green Bay, Wisconsin, the
investigators identified a positive correlation between PCB exposure and birth
weight (Dar et al. 1992). In this study, the estimates of PCB intake may have
been inaccurate because they were based on questionnaire responses subject to
recall. Even if no recall bias existed, uncertainites still exist because the amount
offish eaten in a fish meal varies from individual to individual and the amount
of PCBs and other toxic substances varies within the same species of fish
because of age, size, length, and fat content.
In addition, serum samples were taken early in pregnancy (women with positive
pregnancy tests) within this population, and it has been demonstrated that the
serum half-lives of three different congeners detected in this study ranged from
124 to 338 days (Buhler et al. 1988). Of the 1,112 women in this study, only
106 maternal serum samples were analyzed for PCBs. Because a small number
of maternal serum samples were analyzed the values obtained may not be a true
representation of the entire study population. No additional maternal blood
during pregnancy or fetal cord blood after birth was analyzed for PCB levels to
determine if maternal levels of PCBs varied during pregnancy or to determine
exposure levels to the fetus.
Also during the time of this study, a lot of publicity in the state focused on the
potential for adverse health effects in infants from mothers who consumed
sport-caught fish and on prenatal counseling, including information on toxins
in sport fish. The authors concluded that this publicity probably made the
population more sensitive to fish contamination and therefore, fish consump-
tion during pregnancy may have been less than usual before and during
pregnancy.
In comparing Dar's Wisconsin Maternal and Infant study to the Michigan
Maternal and Infant study, differences exist in study design. The most critical
difference in methodologies was in the selection of exposed populations. In the
Michigan study, the investigators selectively chose individuals who consumed
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large amounts of Lake Michigan fish, (i.e., populations that would have very
high PCB exposure). In comparison the participants in the Wisconsin cohort
consumed much lower amounts of contaminated fish. When the exposure scores
of the Michigan and Wisconsin studies were compared, only 49 persons would
have qualified for Michigan's exposed group. Of the 49 persons who would
have qualified, only 4 participants were estimated to be exposed at the high
Michigan exposure level. Because of the differences in study design, therefore,
the status of human health from exposure to Great Lakes contaminants remains
uncertain.
The Minnesota Ecologic Epidemiologic Study and the New York Ecologic
Epidemiologic Study both hypothesized that populations that live near the lakes
would be potentially exposed to pollutants in air and water, thereby resulting in
adverse reproductive health outcomes (Shuman et al. 1982; and Kagey and Stark
1992). Neither study was able to definitely prove its hypothesis because of the
study limitations.
Ecologic studies have two major limitations: the lack of individual measure-
ments on exposure or outcome and the inability to control for confounding
variables. If individual measurements for exposure or outcome are lacking in
a geographical area as large as the Great Lakes, misclassification is possible. If
exposure was classified incorrectly, it was assumed to be random. This would
tend to underestimate the relationship between residence and outcome. The
second limitation, the inability to control for confounding variables, may have
greatly altered the data outcome. Factors known to affect birth weight, such as
smoking history and alcohol consumption are not recorded on birth certificates,
from which the data for the study were obtained. When data on potential
confounding variables were not recorded, the investigators assumed that they
were distributed equally among exposure areas in the Great Lakes drainage
basin. However, this assumption may not have been correct.
Epidemiologic studies of exposed human populations provide the most con-
vincing evidence of human health effects. Of the three major routes of human
exposure in the Great Lakes — air, drinking water, and fish consumption—fish
consumption is generally thought to present the greatest risk of exposure. The
most direct evidence for adverse human health effects from environmental
pollution is found in a series of studies linking PCB exposure to consumption
of contaminated fish (Fein etal. 1983,1984; Jacobsonetal. 1984a; Jacobsonand
Jacobson 1988). Replicating improving upon, and continuing these types of
epidemiologic studies should provide the most relevant and convincing evi-
dence of the status of human health from exposure to Great Lakes pollutants.
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V. PROGRAM AREAS OF RESEARCH
Given the implications of the association between contaminants in the Great
Lakes and adverse human health outcomes, the following research is being
conducted to clarify the relationships among exposure, contaminant levels in
human biological tissues and fluids, and human health effects. Below are
specific research data gaps that are in the process of being filled, which will
provide necessary information for assessing human risk from exposure to
Great Lakes contaminants.
Characterization of exposure and determination of the profiles and levels
of Great Lakes contaminants in human biologic tissues and fluids
Many studies have identified the presence of numerous toxicants found in a
variety of species in the Great Lakes food chain, including humans (Phillips
andBirchard 1991a,b; Humphrey 1987; Kreiss 1985). Exposure to contami-
nants has been shown to be associated with health effects not only in
individuals consuming fish from the Great Lakes basin but also in their
children.
Exposure to contaminants via consumption of contaminated fish over an
extended period of time allows for continuous exposure that may increase the
potential for adverse human health effects. The persistence and latency of
some of the Great Lakes contaminants have been demonstrated to be
important factors in their ability to cause adverse health effects, such as tumor
promotion, hepatic effects, and reproductive/developmental effects (Humphrey
1988b). Studies are being conducted to:
o Obtain tissues and fluids from exposed and referent populations
for congener - specific analyses. Biological tissues and fluids that
are being analyzed include serum, adipose tissue, cord blood,
placenta, and breast milk in females; serum, adipose tissue, and
seminal fluid in males; and serum and adipose tissue in their
children.
o Determine the levels of accumulated contaminants in human
tissues and fluids over time from established cohort(s) from previous
studies.
o Determine partitioning of contaminants within human tissues
and fluids.
o Determine of release of contaminants into the bloodstream from
body tissues, especially adipose tissue.
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o Determine excretion of contaminants from the body.
o Compare toxicokinetic data among males, females, and their
children.
Identification of sensitive and specific human reproductive end points
Human reproductive effects have been initially assessed in females follow-
ing exposure to Great Lakes contaminats. (Fein et al. 1984; Jacobson and
Jacobson 1988). It is known that Great Lakes contaminants can also
accumulate in male reproductive fluids and organs such as seminal fluid and
testes (Bush et al. 1986; Dougherty et al. 1980). Studies are ongoing to:
o Identify study subjects (male) from a cohort of fish consumers
o Examine sperm morphology, number, viability, and motility in
seminal fluid
o Examine blood samples for circulating levels of Follicle Stimulating
Hormone, Leutenizing Hormone, and testosterone.
o Correlate body burdens of contaminants with levels of hormonal
activity linked to adverse reproductive health outcomes.
Determination of the short- and long-term risk(s) of adverse health effects
in the children of exposed parents
Growth retardation, decreased birth weights, and neurological effects have
been observed in the offspring of mothers who consumed contaminated fish
from the Great Lakes region. There are no reports about adverse health
effects in children from paternal exposure to Great Lakes contaminants. The
following research is ongoing to provide some answers regarding parental
exposure and potential health outcomes in their children:
o Comparing profiles of contaminants found in biological tissues
and fluids of parents with the profile found in offspring.
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o Correlating infant exposure history in utero with adverse
health effects.
o Correlating the exposure history of mothers and fathers with
observed adverse developmental effects in their children.
o Evaluating the potential of developmental effects in children due
to paternal exposure.
o Investigating transgenerational effects in children.
Feasibility of Establishing registries and/or surveillance cohorts in the
Great Lakes
Currently, there are no registries of individuals exposed to multiple contami-
nants in the Great Lakes region via fish consumption. Cohorts of populations
considered at risk because of current or past dietary patterns of consumption
of contaminated fish from the Great Lakes exist, feasibility analyses are
ongoing to:
Identify subgroups of particular concern such as pregnant females,
nursing females, fetuses and nursing infants of mothers who con-
sume contaminated fish, infants and children, and subsistence
anglers including Native Americans, the urban poor, and the elderly.
Conduct (transgenerational) epidemiologic studies in identified fish-
consuming populations from the Great Lakes region with emphasis
on disease and symptoms.
Conduct (transgenerational) exposure studies using available moni-
toring methods to assess mixtures of contaminants present in the
cohorts.
Establish a database that can provide prospective information about
people's health status for use in future health studies, including the
use of validated biomarkers.
This ongoing research will provide information required for ATSDR and
state agencies to perform public health assessments of people at risk from
exposure to hazardous substances. In addition, this research will provide
valuable information for the assessment of human risk from simultaneous
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exposure to chemical mixtures in the Great Lakes region. The results of these
research projects will extend current knowledge of the effects of Great Lakes
contaminants on human reproduction, especially male reproductive toxicity,
and resulting adverse health effects in the children of exposed individuals.
This research will also establish new cohorts for research efforts in the Great
Lakes basin that can be used for future investigation of other human
toxicological end points. Finally, this research will also eliminate a number
of uncertainties in the risk assessment process, thereby providing direction for
future research.
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ATSDR
RESEARCH PROGRAM
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VI. ATSDR GREAT LAKES HEALTH EFFECTS RESEARCH
STRATEGY
GOALS
The goals of the ATSDR Great Lakes Human Health Effects Research Program
is to identify human populations residing in the Great Lakes basin who may be
at risk due to contact with chemical contaminants present in one or more of the
Great Lakes and to prevent any adverse health effects.
STRATEGY
The following strategy will be implemented in support of ATSDR's Great
Lakes Human Health Effects Research Program goal. The strategy is built upon
the five traditional elements of disease prevention:
1. Identification of patterns of morbidity and mortality (through use of
surveillance systems, exposure registries, and reports from state/
local health agencies)
2. Evaluation of causal factors accountable for the observed pattern of
morbidity or mortality (through epidemiologic investigations and
experimental research)
3. Control of the factors found or thought to be accountable for the
observed morbidity or mortality (through health advisories, regula-
tory actions, medical interventions)
4. Dissemination of information about the identification, evaluation,
and control of the observed pattern of morbidity/mortality (through
informational resources such as state programs, local advisories,
media, and publications)
5. Infrastructure to support the identification, evaluation, control, and
dissemination elements of disease prevention (through institutional
mechanisms that involve staffing, budgets, and organizational ar-
rangements)
In order to identify human populations that may be at special risk for adverse
health effects, particularly from consumption of sport fish from the Great
Lakes, ATSDR will fund research to better characterize exposure pathways,
associated body burdens, and potential human health effects from exposure to
persistent toxic substances in the Great Lakes basin with emphasis on vulner-
able populations. ATSDR will also coordinate with state and local health
agencies in the Great Lakes Basin to obtain any surveillance data, reports of
morbidity, and advice that might be relevant to identifying populations at health
risk.
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Once human populations thought to be at risk for adverse health effects are
identified, the health status in select populations will be evaluated and
toxicological studies conducted, where indicated. Adverse health effects
would lead to control actions that would include communication of health
advice from ATSDR to state/local agencies, possible establishment of surveil-
lance or registries of high-risk populations, and advice to regulatory agencies
for their consideration. ATSDR will draw upon its Division of Health
Education to coordinate the dissemination of results from its research program
that pertain to identifying at risk populations in the Great Lakes basin and
actions to prevent any adverse health effects. These activities will include
physician education, assistance to state and local health departments, and any
necessary preventive programs to protect human health.
Also in support of this research program, the Agency will rely on the advice
from the Agency's Board of Scientific Counselors, through its Workgroup on
Great Lakes Human Health Effects Research.
This strategy provides an overall framework for:
o Coordination and implementation of needed health research;
o Utilization of findings from this program and other relevant efforts; and
o Development of integrated approaches to decrease exposures and mi-
tigate toxicity throughout the basin.
SCOPE
There are a wide range of contaminants present in the Great Lakes. These
contaminants include persistent toxic substances, such as; PCBs, DDT and its
metabolites, dieldrin, toxaphene, mirex, methylmercury, benzo[a]pyrene,
hexachlorobenzene, furans, dioxins, and alkylated lead. These persistent
toxic substances bioaccumulate in the food chain, with humans being among
the ultimate recipients. Although more research is needed, preliminary
studies indicate that humans are at risk. In particular, Native Americans, sport
anglers, fetuses and nursing infants of mothers who consume contaminated
Great Lakes fish may have a potentially higher risk of long-term adverse
effects resulting from exposure to these contaminants. Accordingly, ATSDR's
research program targets these high risk populations, and the human health
effects associated with consumption of contaminated fish.
As an outgrowth of this research and other relevant human health effects
findings, EPA will coordinate with appropriate federal, state, and local
agencies, universities, and other organizations in the development and imple-
mentation of preventative programs to protect human health. Coordination
with Federal agencies will include the Food and Drug Administration (FDA),
23 ' Great Lakes Human Health Effects Report
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the Centers for Disease Control and Prevention (CDC), the National Insti-
tutes of Health (NIH), and the Indian Health Service (IHS), as well as the
Agency for Toxic Substances and Disease Registry (ATSDR). EPA will also
continue to work closely with ATSDR and other organizations that have
significant programs relevant to the Great Lakes basin, including the Inter-
national Joint Commission, the Great Lakes Health Effects Program of
Canada. (Figure 1).
In addition, ATSDR will convene forums, as needed, to facilitate the
exchange of research findings and ideas. These forums will include publicly
announced meetings of ATSDR's workgroup of its Board of Scientific
Counselors to review its research program in terms of progress, scope, and
overall direction.
Based on findings from these and other efforts, ATSDR, with other organi-
zations will develop appropriate preventive strategies to avert exposures,
mitigate toxicity, and to increase public awareness about the health effects of
persistent toxic substances in the Great Lakes basin. Specifically, results
from these research efforts will be forwarded to EPA's Great Lakes National
Program Office, which coordinates with all U.S. Great Lakes programs.
Research Program Objectives
The objectives of this research program are to:
o Build upon and extend the results from past and ongoing research
o Develop information databases and/or research methodology that
will provide long-term benefits to the human health effects research
efforts in the Great Lakes basin
o Provide direction for future health effects research
o Provide health information to stateAocal health officials, the con-
cerned public, and their medical health care professionals
o In concert with state/local health officials increase public awareness
regarding the potential health implications of toxic pollution in
Great Lakes basin
o Coordinate, as necessary, with relevant U.S. Public Health Service
research programs and activities, including those of FDA, CDC,
NIH, and IHS, as well as the EPA, and State and local health de-
partments to ameliorate adverse public health impacts of persistent
toxic substances in the Great Lakes basin.
These objectives listed for the ATSDR Great Lakes Human Health Effects
Research Program are subject to, and may change as additional needs become
evident or when new information becomes available.
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Figure 1
ORGANIZATIONS AT WORK IN THE GREAT LAKES
KEY
MOE Ministry of Environment
HERL Health Effects Research Laboratory (EPA)
FDA Food and Drug Administration
IHS Indian Health Service
NIH National Institute of Health
USEPA United States Environmental Protection Agency
GLNPO Great Lakes National Program Office
IJC International Joint Commission
CDC Centers for Disease Control and Prevention
25
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APPROACH
ATSDR is implementing this effort through a combination of both intramural
and extramural research efforts. For fiscal year 1992, ATSDR announced a $2
million dollar grant program with the states of the Great Lakes basin and
political subdivisions thereof to assess the potential impact on human health
of fish consumption in the Great Lakes region. Eligible applicants were the
Great Lakes states which include Illinois, Indiana, Michigan, Minnesota,
Ohio, Pennsylvania, New York and Wisconsin; and political subdivisions
thereof, which may include state universities, State colleges, State research
institutions, state and local health departments, and federally recognized
Native American Tribes. Collaborative efforts among these potential appli-
cants were encouraged and these efforts were continued and extended in fiscal
years 1993 and 1994.
Program Areas of Research
General areas of research pursued were based on the research needs identified
previously in this report; they included but were not limited to:
o Characterizing exposure and determining the profiles and levels of
Great Lakes contaminants in biologic tissues and fluids in high risk
populations;
o Identifying sensitive and specific human reproductive and develop-
mental end points and correlating them with exposure to Great Lakes
contaminants;
o Determining the short and long-term risk(s) of adverse health effects
in progeny that result from parental exposure to Great Lakes contami-
nants;
o Investigating the feasibility of establishing registries and/or surveil-
lance cohorts in the Great Lakes basin;
o Establishing a chemical mixture database with emphasis on tissue and
blood levels in order to identify new cohorts, conduct surveillance
and health effects studies, and establish registries and/or surveillance
cohorts.
Also during fiscal year 1992, ATSDR initiated development of a technical
information network on human health effects resulting from the consumption
of contaminated fish in the Great Lakes basin. In addition to serving as an
information and communication network for ATSDR and those involved in
health effects research it is anticipated that this effort will serve as a resource
to state and local health departments to increase public awareness regarding
potential human health implications associated with toxic pollutants in the
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Great Lakes basin. The peer-reviewed information database available for the
persistent toxic chemicals will be further assessed through ATSDR's toxico-
logical profiles on specific chemicals. Data gaps and research needs identified
in the profiles for specific chemicals will be forwarded to ATSDR's Program
of Substance Specific Applied Research.
SUMMARY
This research program can be viewed in the context of a generic inter-related
tiered model of applied research (Figure 2) intended to delineate the relation-
ships among contaminant levels in the ambient environment, exposure path-
ways, tissue levels, and potential human health impacts. These research
activities and associated findings will be coupled to appropriate public health
activities designed to prevent adverse health effects in vulnerable populations.
Related activities will include public health assessment and consultations,
health studies and health education and other assistance to state and local health
officials as appropriate. Collectively, these efforts will more fully characterize
the potential health effects associated with exposure to persistent toxic sub-
stances in the Great Lakes basin and identify necessary actions to interdict
exposures, mitigate toxicity, and increase public awareness regarding the
potential health effects of these substances.
Model for the relationship between exposure to
hazardous substances and disease
Figure 2
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This strategy has been endorsed by the Council of Great Lakes Research
Managers and has been adopted by the International Joint Commission as a
framework for the study of human health and other ecosystem effects in the
Great Lakes basin. Similarly, this research model can be applied to other
ecosystems across the United States, e.g., Chesapeake Bay, Puget Sound
(Seattle, WA) and the Gulf of Mexico (refers to the Gulf states of Alabama,
Florida, Louisiana, Mississippi, and Texas). The Gulf of Mexico, like the Great
Lakes, is another unique and vital ecosystem which over time has developed
significant pollution problems which have affected water quality,
fish, wildlife and humans who reside there.
IMPACT OF ATSDR RESEARCH PROGRAM
The ATSDR Great Lakes Human Health Effects Research Program is designed
to investigate and characterize the association between the consumption of
contaminated Great Lakes fish and short- and long-term harmful health effects
as evidenced by its six objectives. The greatest potential benefit to be realized
from this program most directly impacts the citizens of the Great Lakes basin
who have lived with unanswered health-based questions concerning exposure
to contaminated fish. The program will also benefit communities in other
ecosystems who can apply the results of studies directly (or indirectly) to their
own exposure situations.
As of September 30, 1994, the ATSDR 3-year Great Lakes Human Health
Effects Research Program has:
o Documented Exposure
Exposure to all 11 persistent toxic substances of concern identified by
the International Joint Commission have been investigated in pre-
sumed susceptible populations via fish consumption. These 11 toxic
substances are PCBs, DDT and its metabolites, dieldrin, toxaphene,
mirex, benzo[a]pyrene, hexachlorobenzene, furans, methylmercury,
dioxins, and alkylated lead. Future analyses of these data may indicate
potential exposure to all or some of the 11 persistent toxic substances
and perhaps additional substances. These exposures have been docu-
mented in populations that build upon six existing cohorts in the Great
Lakes region:
The New York Sport Angler Cohort
The Red Cliff Indian Tribe Cohort
The Mohawk Indian Tribe Cohort
The Michigan Sport Angler Cohort
The New York Breast-feeding Mothers Cohort
The Wisconsin Sport Angler Cohort, and
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two new cohorts:
Michigan Cohort - reproductive age men and women (18-34)
Illinois Cohort - African American women.
o Characterized Exposure
The profile and levels (i.e., body burden) of Great Lakes contaminants
within presumed susceptible populations have been investigated.
These preliminary body burden data may suggest the need for further
investigation of population or lake-specific parameters of exposure.
o Identified Populations at Higher Risk
The preliminary data obtained from the documentation and character-
ization of exposure has served to help identify susceptible populations
who are at greater risk for potential health effects due to their contact
with Great Lakes contaminants. These populations include sport
anglers, Native Americans, urban poor, the elderly, reproductive-aged
men and women, pregnant women, fetuses and nursing infants of
pregnant or lactating women who consumed contaminated fish.
o Identified Potential Human Health Effects
The initial health-based data generated from surveys, review of hospi-
tal records, and clinical follow-ups have been evaluated to help
determine human health effects of concern (i.e., reproductive, develop-
mental, neurobehavioral, immunologic, and endocrinologic) in higher
risk populations.
o Identified Other Potential Pathways of Exposure
Research data from the ATSDR 3-year program has provided a basis
to identify other pathways of exposure to Great Lakes contaminants,
(e.g., occupational, residential, Great Lakes wildlife, and other envi-
ronmental sources).
o Identified Other Potential Geographical "Areas of Concern"
There are currently 43 geographical areas of concern that have been
identified in the Great Lakes basin. Data from the ATSDR 3-year
research program has identified common parameters that link these
susceptible population; facilitating the identification and subsequent
investigation of other potential Areas(s) of Concern.
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o Developed Directions and New Methodology for Future Human Health
Research
Preliminary analyses of available data from the ATSDR Great Lakes
research program has been used to direct research efforts focussing on
higher risk populations in the Great Lakes basin. For example, new
analytical methodology is being developed that will allow greater
sensitivity for detecting low levels of Great Lakes contaminants in
human biological tissues and fluids and other environmental media,
(e.g., Great Lakes fish).
o Provided Health Information
ATSDR has pursued continuing (or new) cooperative agreements or
grants with all eight state health departments and related Federal
agencies in the Great Lakes to facilitate community outreach programs
that educate and increase the public's awareness of the pollution
problem in the Great Lakes. ATSDR will pursue similar activities with
other federal agencies and health care-professionals throughout the
Great Lakes.
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TIMELINE
MILESTONES
Letter to Potential Funding Applicants
in the Great Lakes
Establishment of Subcommittee to ATSDR
Board of Scientific Counselors (BSC)
Presolicitation Notice for Great Lakes
Research Applications
Public Meeting
First Program Announcement
Receipt of Applications
Complete 3-Tiered Peer Review
Award 9 Great Lakes Grants
BSC Workgroup Review
Letter to Potential Applicants
Second Program Announcement
Solicitation for new Application for Grants
Award 1 New Great Lakes Grant
Great Lakes Symposium
Funding of 10 Continuation Awards for FY 1994
BSC Workgroup Review
DATE
02/20/92
03/27/92
04/28/92
04/29/92
06/11/92
07/15/92
08/19/92
09/30/92
11/5/92
06/14/93
07/29/93
1995
09/24/93
05/04/94
08/31/94
04/21/95
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VII. ATSDR GREAT LAKES RESEARCH PROJECTS
SYNOPSES OF FUNDED AWARDS
During fiscal year 1992, ATSDR funded nine research grants to study the
potential adverse human health effects from consumption of contaminated
Great Lakes fish. These studies included eight epidemiologic investigations
in presumed susceptible populations, (i.e., Native Americans, sport anglers,
the urban poor, pregnant women, and fetuses and nursing infants of mothers
who consume contaminated Great Lakes fish). The ninth study focused on
developing more sensitive methods to detect persistent Great Lakes contami-
nants in human biologic tissues and fluids.
The 10 grants funded by ATSDR for fiscal year 1993 included the nine
continuation awards funded in 1992 and one new award that established an
interlaboratory-based, quality assurance and quality control program for the
ATSDR Great Lakes Research Program.
The 10 awarded grants of this program represent 7 out of the 8 Great Lakes
States (excluding Pennsylvania), which include state academic institutions
and 6 state health departments. All five of the Great Lakes - Michigan,
Superior, Ontario, Erie, and Huron are sites of study in this research program.
Additionally, all 11 persistent toxic substances,(i.e., polychlorinated biphe-
nyls, dioxins, toxaphene, benzo[a]pyrene, mirex, DDT, dieldrin,
hexachlorobenzene, furans, alkylated lead, and methylmercury), are being
studied in the identified presumed susceptible populations.
These 10 grants represent two types of studies: (1) epidemiologic investiga-
tions to characterize exposure and human health outcomes in susceptible
populations, with emphasis on expanding existing cohort studies and identi-
fying and evaluating sensitive human health effects, (i.e., reproductive and
developmental effects) and (2) environmental studies to characterize expo-
sure pathways for persistent toxic substances in the Great Lakes basin, with
emphasis on developing new methods and assessing the relationship between
exposure pathways and body burden.
There are no results of these studies in this report. Results present from all
studies will be summarized in a supplement to this document, upon comple-
tion of the studies.
PCB AND DDE EXPOSURE AMONG NATIVE AMERICAN MEN
FROM CONTAMINATED GREAT LAKES FISH AND WILDLIFE
The Mohawk Nation at Akwesasne is a Native American community of about
10,000 people located along the St. Lawrence River in New York, Ontario,
and Quebec. The St. Lawrence River serves as a potential conduit for the
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industrial pollution that originates throughout the Great Lakes. In addition
to this generalized contamination, several industrial sites which serve as
emission point sources for PCBs and several other chlorinated hydrocarbons.
Fish, ducks, and other wildlife that were recently caught near Akwesasne
have been contaminated with a mixture of chemicals, including PCBs,
polychlorinateddibenzofurans,polychlorinateddibenzodioxins,polyaromatic
hydrocarbons, mirex, and DDE, (a breakdown product of DDT). Fish and
wildlife caught near these industrial facilities often have contaminated levels
that exceed the U.S. Food and Drug Administration tolerance limits for PCB s
in fish or poultry.
The New York State Department of Health (NYSDOH) conducted research
on nursing women and their infants from 1986 to 1991 because of concerns
about potential PCB contamination of breast milk. The project was originally
requested by one of the study co-investigators, a Mohawk midwife. The
initial results for the years 1986 to 1990 showed that two specific types of
exposure placed a subgroup of Mohawk women at risk of developing higher
PCB body burdens: a history of local fish consumption among subjects who
gave birth in 1986 to 1989, and residence on Cornwall Island, an area very
close to and predominantly downwind from the industrial facilities. This
residential association was independent offish consumption and was persis-
tent throughout the study period, including 1990.
The major purpose of this ATSDR-supported research project was to
determine if similar associations existed between dietary and residential
exposures to PCBs and DDE and individual body burdens in Mohawk men.
This was the first time that differences in dietary exposure, body burdens, and
potential adverse health effects due to various PCB congeners in men and
women from the same population was explored. In other fish-eating popula-
tions, adult men have consistently shown higher serum levels of PCBs and
DDE (or DDT) than adult women and children. This also appears to be the
first time that the PCB congener pattern for a hazardous waste site as a point
source will be directly compared with congener-specific biologic markers of
dose. Adult men, who are husbands or partners of women in the ongoing
NYSDOH study of Mohawk mothers and infants, will be invited to partici-
pate in an interview and to donate blood samples, in order to achieve the
following research objectives:
1. Estimate exposure to total PCBs, 67 PCB congeners, and DDE from
dietary and residential routes. Concentrations of these chemicals
in fish and wildlife, ambient air, drinking water, and soil adjacent to
individual residences will be related to dietary history and residential
history.
2. Correlate these estimates of exposure with the concentrations of
total PCBs, 67 PCB congeners, and DDE in serum. The estimated
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dose for each specific source of contamination will be separately
correlated with contaminant concentrations in serum.
3. Test the ability of total PCBs, 67 congeners, andDDEto induce liver
enzymes, such as cytochrome P-450, in humans. Liver function are
being examined through the caffeine breath test, which is a sensitive,
non-invasive method of assessing enzyme induction, one of the
earliest detectable biological responses to PCBs in laboratory ani-
mals. This test appears promising as a method to detect subtle sub-
clinical effects before the onset of overt clinical symptoms. This
measure will be correlated with serurn PCB and DDE levels in a
selected sample of the Mohawk men.
Exposure estimates based on the reported consumption of locally caught fish
and wildlife and residential histories were correlated with the specific pattern
of PCB congeners found in the serum. Mohawk men and women who report
lower dietary or residential exposures will serve as an internal comparison
group. Biomonitoring may provide several potential benefits to the Mohawk
population. Depending on the results, several actions could be indicated for
individuals, or for the overall community, including provision of advice on
the risks of local fish consumption, provision of advice on the benefits and
risks of breastfeeding, and targeted remediation of contaminated Mohawk
land. Results of this study may also provide some guidance in the overall
strategy for selecting remedial efforts related to the three major emission
sources.
This project is being conducted by Dr. Kelley Ann Brix, (New York State
Department of Health), in collaboration with Drs. Brian Bush, Edward
Fitzgerald, and Syni-an Hwang (State University of New York at Albany);
Ms. Katsi Cook (Research Foundation, State University of New York at
Albany); and Dr. George H. Lambert (Robert Wood Johnson School).
PROGRESS AND PRELIMINARY RESULTS
Individual Exposure Assessment
The husbands or partners of the 97 women in the previous study of Mohawk
women and infants (1986 to 1991) were contacted to participate in this study.
The response rate of the Mohawk men to participate is expected to be greater
than 67 percent. This enthusiastic response is expected because the Mohawk
people are highly concerned about environmental contamination at Akwesasne.
Many Mohawks used to rely heavily on local fish and game for food, adding
practical and economic significance to the issue.
Each male subject was interviewed about his sociodemographic characteris-
tics, height and weight, use of medications, occupational history, residential
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history, cigarette smoking and alcohol use, drinking water source, and diet.
Additionally, each subject will be asked to mark his current and previous
residences, places of employment, and fishing areas on a detailed map of the
reserve. A coding scheme to map these locations has been developed. The
feasibility of using urine for PCB and DDE analysis is currently under
investigation. If it proves to be a viable approach, each man will be asked to
provide a 1.0 liter sample of urine.
Environmental Exposure Assessment
The environmental exposure assessment process includes three types of
tasks. First, existing environmental exposure data from several different
sources need to be compiled, organized, and integrated. Second, research
gaps identified in the existing exposure information will be addressed
through the collection of additional environmental samples. Third, for each
major source of exposure, each individual will be placed into a qualitatively-
ranked exposure category through the development of an environmental
exposure index. Separate indices are being estimated for PCB and DDE
exposure and for dietary intake of local fish and wildlife, other local
foodstuffs, and residential exposure.
Much of the environmental sampling data necessary to develop estimates of
cumulative lifetime exposure can be derived from data on fish, wildlife, soil,
river sediments, and other media collected previously by the NYSDOH the;
Federal and State Superfund-related remedial investigations by the U.S. EPA
or its consultants, the New York State Department of Environmental Conser-
vation, the Mohawk Nation, and the Canadian federal and provincial govern-
ments. A complete listing of all available relevant environmental sampling
documents has been compiled. These documents have already been summa-
rized and are now being computerized into one database. The organization
and integration of this very large database will be complicated by the diversity
of media (e.g., fish or soil).
These environmental data will be mapped using MAPINFO, which is a
geographical information system. The available map data for the U.S. side
of Akwesasne have already been computerized using information from the
U.S. Geological Survey, the N.Y. State Department of Transportation, and
other resources. These computerized U.S. base maps will be used to plot the
locations of Mohawk residences, fishing areas, soil samples, and so on.
To fill information gaps, additional air, water, and soil sampling will be
targeted to locations that are relevant to the dietary and residential activities
of Mohawk men and women with higher PCB body burdens and internal
controls with lower PCB body burdens. The residential histories of the
individuals have been evaluated for their proximity to areas of known soil
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contamination and for their potential for atmospheric transport downwind of the
General Motors, Alcoa, or Reynolds sites. Geographic patterns will be
determined, including residential clustering of several individuals with high
PCB levels.
Additional air sampling is being conducted by the environmental technician and
Mohawk environmental scientists. Five air monitoring stations, installed in
February 1993, were designed to obtain continuous 28 day samples of ambient
air for PCB analysis. Samples were collected for at least 12 months. Three
monitoring locations were selected on Cornwall Island, and one location was
selected on Raquette Point. All four locations were selected due to their close
proximity to General Motors. A control monitoring station was chosen several
miles upwind from the three factories. Samples of residential soil, drinking
water, and locally grown foodstuffs were collected in the spring and summer.
Laboratory Analyses for PCBs and DDE in Human and Environmental Samples
This work is being conducted in Dr. Brian Bush's laboratory at Wadsworth
Center for Laboratories and Research in the NYSDOH. Dr. Bush directs the
laboratory that performs the PCB and DDE analyses. He has many years of
experience in the congener-specific analyses of PCBs.
Caffeine Breath Test
The caffeine breath test (CBT) will be used to assess liver enzyme induction in
Mohawk men. The first group of 20 eligible Mohawk men were tested in late
summer 1993. An additional 20 subjects will follow.
Summary of Changes Made in the Program
In general, the methods proposed for this study have been followed closely. The
only significant modification needed is the addition of one staff position for an
assistant research scientist, in order to increase the number of male Mohawk
participants.
There are two important reasons to include the husbands or partners of the
Mohawk women who were previous participants in this study (1986 to 1991).
First, this will greatly increase the sample size for the male subjects. Second,
the PCB concentrations found in the breast milk of the earlier women were
significantly higher than the concentrations found in women in 1992. It is likely
that the partners of the earlier women would also have higher PCB body burdens
as well, due to similarities in consumption of local fish and residential
exposures.
Summary of Participants' Evaluation of the Program
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Although it is too early to determine the male Mohawks' evaluation of this
project, the community response to the women's project has always been
favorable. Indeed, one of the most important reasons that the men's project
was proposed is requests by both men and women at Akwesasne that the study
be extended to include men.
When the analyses of the biologic specimens are completed, the results of the
PCB and DDE levels are mailed to each participant and to his or her physician,
along with interpretations. Consultations with the principal investigator, who
is a physician epidemiologist, is offered in the notification letters, occasion-
ally, subjects or their physicians call with questions about the clinical
significance of their body burdens.
In 1992, a planned series of annual public meetings was initiated to explain
the results of the study and to provide an interpretation in lay language. All
the participants, to date, were individually invited, as well as other members
of the community. This public meeting also served as a forum for community
input before the initiation of a new stage in the study (in this case, the caffeine
breath test). The directors of the two Environmental Divisions at Akwesasne,
Mr. Ken Jock and Mr. Henry Lickers, also provided important contextual
information by discussing the progress in negotiations over the remediation
at the three industrial facilities. The community was supportive of the
NYSDOH study, as demonstrated by the positive response at the public
meeting. Another public meeting with a similar agenda is planned for 1993.
Other systematic attempts to publicize study results have included a press
conference and individual contacts with reporters at the tribal newspaper,
other newspapers, radio and television stations. Continued community
support will be encouraged by providing opportunities for public participa-
tion in the planning and implementation of the study and in the timely
reporting of results.
Planned Activities for Year II
The four major activities initiated in 1992 were expanded: the recruitment
and interviewing of study participants, the environmental exposure assess-
ment, the laboratory analyses for PCBs and DDE, and the caffeine breath test.
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AN ASSESSMENT OF A HUMAN POPULATION AT RISK: THE
IMPACT OF CONSUMING CONTAMINATED GREAT LAKES FISH
ON NATIVE AMERICAN COMMUNITIES
The Ojibway (Chippewa) people of the upper Great Lakes are important
candidates for study because of their dependence on fish as a food source and
as an economic base. This project involves the study of fish consumption
habits, body burdens, and neurobehavioral effects of several Ojibway tribal
bands that reside in the Lake Superior region. The results of these studies are
expected to indicate whether significant exposure warrants continued
epidemiologic studies or alterations in lifestyles of the community members.
The targeted reservations have a well documented history of fishing cultures,
including subsistence and commercial fishing. Extra fish are distributed
among crew members and the extended family for labor compensation as part
of cultural ritual and tradition. Therefore, the effects of consuming contami-
nated fish are shared by nearly all tribal members.
The objectives of this research program include:
1. Survey adult tribal members and/or their spouses regarding past and
current dietary practices, demographic characteristics, fish con-
sumption, and environmental risk perception. Up to 300 tribal
members at each of six reservations v/ill be surveyed.
2. Collect biologic samples of blood and hair for contaminant analyses
from 50 adults per site. The contaminant data will be used to study
the correlation between reported fish consumption, known fish
contaminants, and human body burdens. The two primary contami-
nants of concern are polychlorinated biphenyls (PCBs) and methyl-
mercury, which are both known to be developmental neurotoxicants.
However, the biologic samples will be screened for other organo-
chlorines, such as DDT, dieldrin, and chlordane.
3. Conduct neurobehavioral assessment on the individuals providing
biologic samples because of the known relationship between many
fish contaminants and neurologic dysfunction. In this researcher's
earlier work with the Red Cliff tribe, there was a significant although
minor, decrement in grooved peg board performance. Such a
disturbance in hand-eye coordination and psychomotor abilities can
be the result of methylmercury or PCS exposures.
4. Collect representative species offish used in diets from each location
for chemical analysis. Local tribal fishermen will provide research-
ers with fish fillets to be analyzed and used later in rodent feeding
bioassays. The fish species selected from each site will depend upon
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the data reported in the fish consumption questionnaires and on the
recommendations of tribal fishery authorities. In western Lake
Superior, lake trout is the predominant species of interest, whereas
from eastern Lake Superior, northern Lake Michigan, and western
Lake Huron, white-fish is the most important species.
5. Conduct neurotoxicity testing in animals. Diets consisting of a
composite sample offish (30 percent) from each site will be given to
laboratory rodents. The adult rats and their offspring will be tested
in several behavioral protocols to determine whether the contami-
nated fish diets are capable of causing adverse effects. Rodent studies
using these fish will be particularly useful to help define critical
exposure levels, not only for individual contaminants, but also for
combination of environmental toxicants that are likely to occur in
actual human consumption. Previous studies demonstrated that
such feeding protocols result in elevated contaminants in the brain
and adverse reproductive and behavioral effects.
The primary benefit of the Ojibway health study to the participating individu-
als and the tribal communities is the identification of contaminants in their
diets. Also, identifying ways to alter their cooking methods and determining
the fish species, fish size, and fishing locations that have fewer contaminants
will help the tribes to reduce their exposure risks.
Another important benefit to the participating community is the educational
outreach portion of the project. For this part of the project, exhibits describing
the general problems with contaminants in the Great Lakes and specific
information on fish consumption health advisories are provided to all age
groups at community health fairs, pow-wows, and school- sponsored events.
These displays increase public awareness of contaminant problems in the
Great Lakes, as well as to help provide excellent opportunities to recruit
subjects.
This research project is being conducted by Dr. John Dellinger at the Medical
College of Wisconsin in Milwaukee Mr. Larry Brooke at the University of
Wisconsin-Superior, Lake Superior Research Institute; Dr. Richard Hoffman
at the University of Minnesota at Duluth; and Dr. Marty Kanarek at the
University of Wisconsin at Madison.
PROGRESS AND PRELIMINARY RESULTS
Reservation Activities - Year One
During the first part of funded year, various components of the project were
initiated, tribal approvals were obtained, and preliminary visits were made.
The investigator established contact with five reservations (Grand Portage,
39 Great Lakes Human Health Effects Report
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MN, Bad River WI, Keweenaw Bay, MI, Bay Mills, MI, and Saulte Ste
Marie, MI). Four of the five elected to allow tribal members to participate in
the study. Only Saulte Ste Marie declined, and they expressed a willingness
for later participation. In most cases, two visits to each site were necessary
to obtain tribal council approval (one preliminary and then attendence at a
council meeting). The tribal council members usually asked very detailed
questions and were most concerned with the impact the study could have on
their commercial fishery activities. They indicated a need to hear the results
of the study prior to public release. The preliminary visits were also used to
attempt to arrange for local tribal liaisons to assist in data collection. The
researchers were successful in making liaison arrangements at Keweenaw
Bay and Bad River. Bay Mills reservation elected to use tribal officials to
collect the data.
Questionnaire Results and Epidemiology
During the summer of 1993, preliminary data were collected at health fairs
and pow-wows. Approximately 200 questionnaires were completed by tribal
members attending the health fairs for all reservations except for Grand
Portage. At Grand Portage, extensive "door-to-door" solicitation by study
investigators was necessary to obtain the data.
Of the approximately 200 surveys, 124 (62%) were completed by females.
The surveys had four sections including fish consumption, fish preparation,
demographics, and risk perception. Preliminary data from the fish consump-
tion and fish preparation sections have been entered for 86 participants,
primarily the Bad River reservation. The respondents reported eating a fish
meal an average of 33.9 days in the last 12 months. The majority of the
respondents consumed walleye (74%) followed by lake trout (56%), smelt
(44%), whitefish (43%), salmon (36%), and perch (34%). The average
number of years reported for consuming Great Lakes fish was 27 with a range
of 2 to 75 years.
Walleye, was the most frequently consumed species reported, was eaten on
average less than once a week, but at least once a month throughout the year.
Two-thirds of the respondents reported that Lake Superior and its tributaries
were the sources of the walleye they consumed. The average length of
walleye that was most commonly eaten was 13 inches. Walleye was most
frequently prepared as fillets (skin-off) and pan or deep fried. Lake Superior
was also the primary source reported for the other most frequently consumed
species — lake trout, whitefish, and salmon. Pan frying and baking were the
most commonly reported cooking methods for all species.
Health Advisories
Awareness of health advisories was assessed by the questionnaire. Twenty-
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nine respondents (46%) were not aware of the fish consumption health
advisories, eight (13%) had read the appropriate advisory, and twenty-six
(41 %) were aware of the advisory. The respondents were most likely to use the
advisory to determine which lakes to fish and how often they should consume
certain species.
Chemical Analyses
Twenty participants provided hair and blood samples for total mercury and
PCB analyses. PCB analyses have not been completed. Preliminary data from
22 individuals have been completed for blood mercury levels; 11 of these
individuals are from Grand Portage, 6 from the township of Baraga on the
Keweenaw Bay reservation, and 5 from the township of L' Anse at Keweenaw
Bay.
The grantee also completed mercury analyses on a number of walleye samples
provided by the Great Lakes Indian Fish and Wildlife Commission (GLIFWC).
These samples included 14 walleye fillets either with or without the skin for
analyses. GLIFWC needed the information to respond to the concerns of
members of the Wisconsin Department of Natural Resources who prefer to
analyze all fish as skin-on fillets. However, our studies and GLIFWC data
indicated that the Native Americans prefer walleye without the skin. The
results are being reported here because of the relevance to the ongoing ATSDR
project. Leaving the skin on the walleye fish appears to result in a 10 percent
dilution of the total mercury and, therefore, leads to underestimation of total
mercury exposure through walleye consumption. All work on this project will
continue to use walleye fillets without skin. For all other species, skin will be
left on only when it is the preferred consumption method for the Native
Americans.
Animal Studies
Because of the reported high consumption of walleye by the Ojibway people,
the grantee decided to have additional walleye samples prepared for a rodent
subchronic feeding study. Forty-eight female Long-Evans hooded laboratory
rats were fed composite fish sample diets (30% of the total diet) of walleye
contaminated with 0.79,0.42, and 0.19 ppm mercury. At this time, preliminary
results indicate no significant behavioral effects in the adult rats. Therefore,
similar diets can be fed to pregnant female rats in a subsequent developmental
protocol for the ATSDR project. In related collaborative efforts, 32 of the 48
rats have been euthanized and are undergoing extensive mercury distribution
analyses using autometallagraphy.
Neuropsychological Testing
Dr. Richard Hoffman, clinical psychologist at the University of Minnesota -
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"'" *£••
Duluth Medical School conducted neuropsychological examinations on 28
tribalmembersfromGrandPortageandKeweenawBay. InJuly 1993,20Grand
Portage tribal members were scheduled for testing, and 11 subjects completed
the testing. Seven female subjects were tested (mean age = 46.6 years, mean
level of education = 12.7 years), and 4 males were tested (mean age=46.3 years,
mean level of education = 15 years). In July and August 1993, 25 Baraga/
L* Anse tribal members were scheduled for human neurobehavioral testing, and
17 completed the testing. Twelve female participants (mean age 36.9 years, a
mean level of education, 13.3 years) were tested. Five males (mean age = 42.2
and mean level of education = 12.4 years) completed testing. It is anticipated
that at least 25 to 30 more subjects from this tribal population will be scheduled
for testing in June 1994. During the winter, Bad River tribal members will be
tested.
Conclusion
In summary, year one provided the grantee with the opportunity to develop
methods, collect approximately 200 completed questionnaires, and evaluate the
effectiveness of the methods. The only significant shortfall in project perfor-
mance has been in the total number of participants recruited at the individual
sites. This has been due to fewer than expected numbers (often the number of
enrolled members does not reflect current residents), the tedious nature of the
questionnaire, and conflicts in participants' schedules. The grantee is sug-
gesting some methodological changes to increase participation (e.g., simplify
the questionnaire, maximize health fair data collections, and simultaneously
collect hair and blood samples from all willing participants)
COGNITIVE AND MOTOR EFFECTS OF PCB EXPOSURE IN
OLDER PEOPLE FROM THE MICHIGAN FISH-EATER COHORT:
EMPHASIS ON THE ROLE OF ORTHO-SUBSTITUTED CONGENERS
Polychlorinated biphenyls (PCBs) are one of the most common chemical
contaminants in aquatic ecosystems, such as the Great Lakes, and consumption
of sport-caught fish from the Great Lakes is a major source of PCB exposure in
humans. Epidemiological investigations and animal studies suggest that PCB
exposure may result in neurobehavioral dysfunction.
Numerous studies have demonstrated age-related decline in cognitive and
motor functioning in humans. Because of this decline, the aged may be at
increased risk for neurological dysfunction following prolonged exposure to
neurotoxicants. This research project is designed to determine the risk of
cognitive and motor dysfunction in aged humans who have consumed large
quantities of PCB-contammated fish from the Great Lakes. In addition to
measuring total PCBs, congener specific glass capillary gas chromatography
and porous carbon chromatography will be used to determine the concentrations
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of the two major classes of PCBs: ort/zo-substituted, non-coplanar PCBs, and
dioxin-like coplanar PCBs. The investigator will access the role of each of
these classes of PCBs in mediating behavioral dysfunction.
In 1980 and 1982, the Michigan Department of Public Health identified and
recruited Great Lakes fish-eaters and non-fisheating controls for a study to
evaluate the relationship between fish consumption and PCB exposure. Data
on serum PCB concentrations, fish consumption patterns, and other health
and diet variables were collected. A recharacterization of the cohorts
undertaken in 1989 and 1991 is completed. These cohorts provide a unique
opportunity to study the neurobehavioral effects of PCB exposure in older
individuals because 62 percent of the study participants are currently 50-79
years of age, and because historical data on fish consumption and PCB body
burden are available for these individuals.
The objectives of the proposed research are given as follows:
1. Complete the recharacterization of the Michigan Department of
Public Health fish-eater and control cohorts.
2. Select a subset of approximately 200 individuals, 100 fish-eaters, and
100 controls, who are 50 to 79 years old and are matched by age and
sex. Demographic data collected during the recharacterization will be
analyzed and used to select the matched groups of fish-eaters and
controls.
3. Evaluate cognitive and motor functioning in aging (50 to 79 years
old) fish-eaters and in their age- and sex-matched, nonfish-eating
controls. The two groups will be tested on a neuropsychological
assessment battery designed to detect subtle deficits in cognitive and
motor functioning. The battery centers on tests that have been
demonstrated to be sensitive in detecting age-related decline in be-
havioral function. Based on PCB studies in animals and in develop-
mentally exposed humans, both cognitive and motor deficits are ex-
pected. Cognitive effects are expected to predominate and tests that
rely on executive functions controlled by the prefrontal cortex are
expected to be most sensitive.
4. Assess the relationship of serum concentrations of PCBs and other
contaminants to behavioral dysfunction. Blood samples will be
collected from participating individuals at the time of the neuro-
psychological assessments and analyzed for PCBs and other con-
taminants via a congener specific method that includes separation and
quantification of coplanar PCBs. The role of three PCB exposure
variables (i.e., total PCBs, total ort/zo-PCBs, and total coplanar PCBs
43 Great Lakes Human Health Effects Report
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[expressed as TEFs]) in mediating behavioral outcomes will then be
assessed. The potential role of other major contaminants present in
Great Lakes fish will also be considered. Based on neurochemical
and behavioral studies in animals, onTzo-substituted PCBs are ex-
pected to be most strongly related to the behavioral outcome mea-
sures.
The proposed study will be the first to evaluate the potential neurobehavioral
effects of PCB exposure in an elderly population. It will also be one of the first
studies to evaluate the relative importance of different classes of PCB
congeners in mediating behavioral effects in humans. The study will take
advantage of an already identified cohort of PCB-exposed and control
individuals for which historical data on PCB exposure are available. The
results should determine whether PCB exposure via consumption of Great
Lakes fish exacerbates or accelerates normal age-related decline in behavioral
functions and which class of congeners, 0rz7zo-substituted or coplanar, are
most important in mediating these effects.
This research project is being conducted by Dr. Susan Schantz (University of
Illinois at Urbana-Champaign) in collaboration with Dr. Harold E.B. Humphrey
(Michigan Department of Public Health); Drs. Ann Sweeney (University of
Texas at Houston) and Dr. Joseph Gardiner and Ms. Donna Gasior (Michigan
State University); Dr. Robert McCaffrey (State University of New York at
Albany).
PROGRESS AND PRELIMINARY RESULTS
Subject Interview
A subject interview has been developed and piloted on age-appropriate pilot
subjects. The interview includes questions on a number of important control
variables that could influence the nervous system end point being assessed in
this study. These include demographic background, alcohol consumption,
tobacco use, prescription and non-prescription drug use, medical history
(including psychiatric illnesses), and activity level. The interview takes
approximately 30 to 40 minutes to complete and are conducted by the subject
examiner at the close of the home visit, after completion of the
neuropsychological assessment battery.
The early piloting study suggested that data on fish consumption and
employment history could be better obtained via separate questionnaires sent
to the subjects prior to the home visit. Thus, separate fish consumption and
employment history questionnaires have been developed and are mailed to the
subject approximately one week before the home visit. The completed
questionnaires are picked up by the subject examiner at the time of the home
visit.
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Computer Programming
The subject interview is currently being computerized by programmers at the
Institute for Public Policy and Social Research (IPPSR) at Michigan State
University. IPPSR specializes in developing and conducting computer
assisted survey research for principal investigators using the CASES (Com-
puter Assisted Survey Execution System) software which they lease from the
University of California at Berkeley. The CASES software is very flexible
and has many advantages over paper and pencil interviewing. Subject
responses being directly entered into the computer rather than written out
long-hand substantially reduces interview time. Furthermore, CASES is
programmed to follow skip patterns imbedded in the interview, automatically
advancing to the next appropriate question. It also incorporates relevant
information as the interview progresses, inserting appropriate names and
numbers obtained in one part of the interview into questions at a later point
in the interview. CASES also searches for inconsistencies in the subject
responses and alerts the interviewer to potential inaccuracies. The completed
instrument will be installed on an IBM 486 notebook computer that will be
taken to the home visits for direct computer entry of interview data. This will
allow more accurate and efficient data collection, and will save considerable
time that the research assistant would otherwise have spent coding and
entering data.
Fish Consumption Questionnaire
The questionnaire has been completed and is currently being piloted on Great
Lakes fishermen and may undergo some changes based on their feedback.
The questionnaire asks about consumption of specific fish species from each
of the four Great Lakes that border on the State of Michigan. The fish species
and the individual lakes and tributaries that are asked about were selected
based on the fish consumption advisories published in the 7992 Michigan
Fishing Guide (Michigan Department of Natural Resources 1992). The
questionnaire also asks about consumption of wild game, fish preparation and
cooking methods, and changes in fish consumption practices as a result of the
advisories. Serving size will be determined at the home visit using fish
models on a standard dinner plate.
The fish consumption questionnaire is based on the questionnaire used by Dr.
John Vena and his colleagues for their ATSDR - funded project and is very
similar in format. It is also very similar to the questionnaires that will be used
in the ATSDR - funded projects by Sweeney and Fischer. This should
facilitate collaboration and comparison of data across these three ATSDR -
funded studies
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Employment History
A short employment history questionnaire are included with the fish con-
sumption questionnaire. The investigator uses employment information to
determine potential occupational exposure to PCBs and other chemicals. An
occupational exposure classification system developed by the National Insti-
tute for Occupational Safety and Health will be used. Each job title will be
assigned a unique classification code. Codes will be obtained for each job
reported in the subject's occupational history. These codes will then be
matched to a database that provides an estimate of PCB exposure for that
particular job, if any, as well as estimates of other chemical exposures
associated with the position.
Neuropsychological Assessment Battery
Piloting of the neuropsychological assessment battery and subject interview
began in mid-March, and will continue through mid-May. On March 31
through April 2, the investigators met and assessed the data from the first three
pilot subjects. Three tests, the finger tapping test, grip strength, and object
assembly, were cut from the battery to reduce its length and make it less tedious
for the subjects. The revised battery has been piloted on five subjects, is
proceeding very smoothly, and takes approximately 2.0 to 2.5 hours to
complete, including a ISminutebreakmid-waythroughthesession. Thesame
five subjects also completed the subject interview. Total assessment time
averaged about 3 hours. Four additional pilot subjects, (male, 51; female, 55;
male, 72; female, 71 yearsofage) were scheduled for testing on May 7,1993,
and May 11, 1993.
Sample Population
As detailed in the original proposal, the subjects to be tested in the current
study will be drawn from the Michigan Department of Public Health's (DPH)
fisheater and non-fisheater cohorts. A listing of identifications (IDs) for all
subjects eligible to participate has been obtained from DPH. The sampling
frame is depicted in the following table.
Sampling Frame
Males Females
Age
50-59
60-69
70+
Total
Fish-eaters
87
47
53
187
Controls
37
32
29
98
Fish-eaters
37
34
29
100
Controls
54
50
39
143
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Selection of Individuals
Within each cell (age x sex x exposure), subjects will be selected in groups of
two using a table of random permutations (Fleiss 1986). Initially, this will
result in the selection of 24 subjects, 2 from each cell, who will then be sent
recruitment letters. This selection and recruitment procedure will be repeated
at 2 month intervals until the desired sample size of 15 to 20 participants in each
cell has been attained. Even allowing for a 25 percent refusal rate, the sampling
frame is large enough to ensure an adequate sample.
Recruitment
Subjects were recruited in or via a two-stage process. Subjects were contacted
initially via a letter from DPH explaining the study and inviting them to
participate. They were asked to check, "Yes, I wish to participate" or "No, I do
not wish to participate" on the enclosed form and return it to DPH in a stamped,
addressed envelope that was included with the mailing. Subjects who respond
"yes" were contacted by phone by the subject examiner to set up a time for the
neuropsychological assessment. Subjects who respond "no" were contacted by
a DPH field worker and asked to complete a short nonparticipant phone
interview in which the subject examiner obtained basic demographic informa-
tion.
Subjects who did not respond to the initial mailing within 2 weeks were
contacted by phone by a DPH field worker and asked to participate. If they
agreed, their names were referred to the examiner for scheduling. If they
decline, the nonparticipant interview was conducted.
ADDITIONAL ACTIVITIES:
Complete Piloting of Assessment Battery
Computer programming of the subject interview has been completed. Piloting
of the neuropsychological assessment battery and interview with study popu-
lation is ongoing.
Subject Assessments
The first set of recruitment letters were mailed during the last week in May
1993, and subject assessments began in early June. Approximately 40
assessments were completed in year 1 of the study. Neuropsychological data
will be coded and entered into a computerized database as they are generated.
1993
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Blood Sampling and Analysis
Within several weeks after subjects completed the neuropsychological assess-
ment battery, a phlebotomist from DPH visited them to obtain blood samples for
residue analysis. The samples were analyzed for PCBs and other contaminants
at the Michigan Department of Public Health. Approximately 100-120 subjects
are anticipated to be assessed during year 2 of the grant.
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THE NEW YORK STATE ANGLER COHORT STUDY: EXPOSURE
CHARACTERIZATION AND REPRODUCTIVE AND DEVELOPMEN-
TAL EFFECTS
Many persistent chemicals, such as poly chlorinated biphenyls (PCBs),
polychlorinated dibenzo-p-dioxin (PCDDs), polychlorinated dibenzo-furans
(PCDFs), the pesticides mirex and dichlorodiphenyl dichloroethene (DDE)
and mercury have found their way into the aquatic life and fish-eating
predators. Because of the persistent nature of these chemicals they will
continue to cycle throughout the ecosystem for a long period of time.
Therefore, the effects of past discharges still exist in the Niagara River, other
tributaries, and Lake Ontario. Also, the potential impact of nonpoint source
pollution, in particular chemicals leaching from hazardous waste dumps
could be significant. The migration of chemicals from these dump sites has
been shown to pollute surrounding groundwater, the Niagara River, and Lake
Ontario. Therefore, Lake Ontario and its sediments will continue to act as a
reservoir for pollutants that biomagnify within the aquatic food chain.
The ultimate recipients for these contaminants are humans who consume the
larger fish, especially the predator species, such as salmon and lake trout, and
bottom feeders, such as carp and catfish. The long-term goal of this ATSDR-
supported project is to characterize the exposure of humans to persistent toxic
chemicals associated with the consumption of Lake Ontario fish and wildlife.
This project will also assess the association for the potential of short- and
long-term health impacts using the New York State Angler Cohort, estab-
lished by the Department of Social and Preventive Medicine, State University
of New York at Buffalo.
The New York State Angler Cohort is a representative sample of New York
State fishing license holders between the ages of 18 and 40 in 16 upstate New
York counties. These counties are near or border Lake Ontario. The cohort is
unique in that fish consumption, medical history, and reproductive history,
as well as other epidemiologic data, has been obtained on 10,799 male anglers
and 6,579 of their wives or partners, as well as on 918 female anglers. Out of
the 7,497 women in the cohort, 2,777 reported having at least one live birth
in the past five years. Both paternal and maternal lifetime consumption offish
from Lake Ontario was obtained for 3,453 children born from 1986 to 1991.
In summary, this is the largest cohort offish-eaters ever assembled; it includes
anglers, their spouses, and children. Because of the size of this population, the
investigators will have adequate statistical power to observe even small
effects.
This ATSDR-supported project focuses on continuing this in-depth study
and follow-up of this very important cohort. A multifaceted research program
has been designed to meet the following major long-term goals:
49 Great Lakes Human Health Effects Report
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1. Characterize the exposure of humans to persistent toxic chemicals
associated with the consumption of Lake Ontario fish and wildlife.
2. Evaluate the relationship between both paternal and maternal expo-
sure to Lake Ontario contaminates and sensitive, specific human
reproductive and developmental end points, encompassing several
reproductive outcomes. Identify retrospective and prospective
cohorts for long-term evaluation.
3. Assess specific subclinical health effects among cohort subgroups
with validated exposure to Lake Ontario contaminants. This will
include assessing lipid and glucose metabolism, porphyrin metabo-
lism, liver function, and thyroid function.
4. Re-evaluate the entire cohort to assess short-term mobility and to
develop measures of health outcomes for prospective follow-up
including reproductive outcomes and chronic disease.
These investigations will help clarify the role between consumption of
contaminated fish and reproductive histories, fetal losses, gestational age-
specific birth weight distributions, and developmental end point among their
offspring. Most importantly, the research project will permit prospective
follow-up of the cohort to assess long-term outcomes, such as diagnoses
among adults of chronic diseases.
This research project is being conducted by Drs. John Vena, Germaine Buck,
Hebe Greizerstein, Paul Kostyniak, James McReynolds, Michael Msall,
James Olson, Luther Robinson, James McReynolds and Maria Zielezny
(State University of New York at Buffalo), in collaboration with Dr. Edward
Fitzgerald (New York State Department of Health), Dr. Lowell Sever
(Batelle Pacific Northwest Laboratories), and Dr. Brian Bush (State Univer-
sity of New York at Albany).
PROGRESS AND PRELIMINARY RESULTS
The above four major goals were subdivided into 8 specific objectives as
follows:
Exposure Characterization
Objective 1:
Objective 1 is to characterize exposure of humans to persistent toxic chemicals
associated with consumption of Lake Ontario fish and wildlife. These
chemicals would include 68 polychlorinated biphenyls, DDT, DDE, mercury,
mirex, hexachlorobenzene, and lead.
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Exposure will be characterized in the following populations: (1) four
subcohorts (A-D) of individuals with various levels of validated exposure to
duck, turtle and Lake Ontario fish; and (2) a fifth subgroup consisting of a new
cohort of lactating women.
Cohort A Anglers with the Highest Consumption of Ducks. Turtle, and Lake
Ontario Fish
The investigators decided to concentrate on attempting to quantify exposures
among the highest consumers, with the following groups selected for this
cohort: 30 anglers who consume high amounts of duck, fish and turtle, 30
anglers who consume high amounts of both duck and fish, and approximately
25 anglers who consumed the highest quantity of turtle regardless of their
consumption offish and duck. Levels of contaminants in these subgroups will
be compared to cohorts B, C, and D individuals who consume high, interme-
diate, or zero amounts of Lake Ontario fish, respectively, but no consumption
of duck or turtle. The sample sizes of the subgroups (N=30) will provide
adequate statistical power to detect mean differences across any two groups as
small as .72 of one standard deviation. These estimates are the minimum power
for these given sample sizes. Power is likely to increase with other analytic
methods such a multiple regression with adjustment for other variables.
Specific procedures for contacting, obtaining the supplemental questionnaire
information, and sample processing have been worked out and the subcohorts
have been identified. Sample collection is awaiting finalization of the analytic
procedures. The investigator has already contacted anglers in cohort A for the
study.
Cohorts B. C. and D with High. Intermediate, and Zero Lake Ontario Fish
Consumption
The investigator originally proposed to study 30 individuals (15 men and 15
women) with validated exposure by quantification of the two PCB isomers 138
and 153 among each of the exposure categories. It is likely that because the
exposures are validated, exposure groups will be selected with precision. Due
to comments of the reviewers, all cohorts have been increased to 30 men and
30 women in each group. This increased sample size will allow the investigator
to assess any potential interactions such as differences by sex.
Cohort E: New Cohort of Lactating Woman (N=100)
The investigator proposed to characterize exposures among 50 lactating
women with high consumption of fish and other wildlife and 50 lactating
women with no reported consumption. Collection of breast milk from lactating
women is well underway in conjunction with meeting the requirements of
Objective 6: Follow-up of the planned pregnancy group. A supplementary
questionnaire was developed and pre-tested; each woman providing a breast
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milk sample fills out this questionnaire. Once women are identified on the
telephone interview, a detailed procedure for obtaining the breast milk sample and
supplementary epidemiological information has been worked out. Forty-four
women who are currently breast feeding have been identified to date and 32 have
given a breast milk sample, which is 73% of those lactating. Of the 104 women
identified as currently being pregnant, 44% agreed to give a milk sample with an
additional 13 % unsure about giving a milk sample but are willing to be contacted
at a later time. Therefore, it is quite likely that more than 100 breast milk samples
will be collected.
Analytical Procedures
Analytic procedures for PCB congeners, hexochlorbenzene, DDE, and mirex as
well as the QA/QC methods and data reporting, have been carefully worked out
with the collaboration of Dr. Brian Bush. QA/QC samples have been sent to Dr.
Bush's laboratory.
Selective Measurement of Methylmercury in Hair
A new method for measuring methylmercury in hair has been developed. This
technique employs a sequential analysis in the same aliquot of a pre-digested hair
sample using cold vapor atomic absorption spectrophotometer. When applied to
hair samples, recoveries of organic mercury using this method were greater than
90%. QC samples and standards were approximately 95% reproducible. This
methods gives a lower limit of detection of 1 ng of organic mercury, and is linear
up to over 40 ng of Hg per sample aliquot. The grantee will employ this new
methodology for the analysis of mercury in hah-.
Objective 2:
Characterize Exposure to Polychlorinated Dibenzo-P-Dioxins (PCDDs) and
Polychlorinated Dibenzofurans (PCDFs) in Individuals with High and No
Reported Validated Exposure
Preliminary discussions have been made with Dr. O' Keefe at the New York State
Health Department regarding these procedures. The individuals forthis study will
be identified in year two.
Objective 3:
Assess Specific Subclinical Health Effects in a Cohort with High and Intermediate
Exposure to Lake Ontario Contaminates and a Control Group with No Validated
Exposure
The assessment of specific subclinical health effects will not be conducted until
late in year 2. The subcohorts B (high), C (intermediate), and D (zero consump-
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tion), of Lake Ontario fish identified for these tests will be clearly validated by the
end of year two. The additional time will also allow the grantee to increase the
subcohort size in each group to obtain greater statistical power for analyses.
Objective 4:
Assess Human Reproductive End Points in all Members of the Cohort, Including
Anglers and Wives, and Partners of Male Anglers Procedures to meet this study
objective are well under way. Detailed proposals for access to the birth
registration, fetal death registration, and other record systems of the New York
State Department of Health have been prepared, submitted, and approved by the
New York State Department of Health Institutional Review Board (IRB).
Identifying information for all male and female anglers and the wives and partners
of male anglers including birth dates, full names, addresses, and social security
numbers have been prepared on a data tape. It is projected that by the end of year
1, the updated information from these registries will be available for use in year
2 of the grant when the investigator will correlate validated consumption measures
with the outcome data.
Objective 5: Intrauterine Growth and Developmental End Points
Objective 5 is to collect hospital delivery records for infants born to female anglers
or wives or partners of male anglers included in the New York State Angler Cohort
(N=3,658). Hospital delivery records will be sought for infants born to cohort
members between June 1,1986, and June 1,1991. The hospital of birth has been
contacted for all pregnancies occurring during this 5-year study period. Of these
births, 135 or 4% occurred outside New York State, and 199 or 5% has missing
birthplace data. Birth certificates will enable the investigator to identify these
missing hospitals. The remaining live births (N=3,324) occurred in 94 different
hospitals across New York State. A specific request to obtain data from the
hospital were submitted to the New York State Institutional Review Board (IRB).
While awaiting final IRB approval, a standardized data abstraction form was
developed and reviewed by the clinical consultants. The abstraction form was pre-
tested and finalized and is now ready to implement. The investigator also prepared
a list of births from each hospital and developed letters of contact for hospitals in
the different areas of the state; they were stratified by number of records per
hospital and geographic region. It is projected that considerable effort will be
required to abstract the hospital and delivery records to obtain the critical data
needed to assess intrauterine growth and establish developmental end points for the
cohort.
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Objective 6: Surveillance of Planned Pregnancy Group
The initial angler cohort survey identified 3,005 women who reported that they
were either planning another pregnancy or that they were undecided about
future childbearing. In year 1, a brief telephone interview was initiated to
update the reproductive histories and to establish them as a cohort for future
surveillance. The detailed telephone interview was developed, pilot tested and
has already been implemented. During the first six months of the grant, 784
completed interviews have been logged; only 13 women who were contacted
have refused (<2% refusal). Of the 784 women interviewed, 400 have become
pregnant since 1991 (51%), 104 women reported to be currently pregnant and
87 women were currently trying to become pregnant (11% of those inter-
viewed). Seventeen percent of those interviewed have decided not to have any
more children. Interviewing of this subcohort will continue in year 1, but the
rate of interview completion will decline once the nurse research assistants
begin medical record abstracting.
Objective 7: Comprehensive Follow-Up of Cohort
Objective 7 is to resurvey the entire cohort in the third year of the grant and,
therefore, is not applicable at this time.
Objective 8:
Objective 8 is dissemination of findings. As findings are generated during year
2 and at the end of year 1, they will be disseminated to the populations at risk,
through presentations to ATSDR at the annual progress meetings, reports to
ATSDR, and subsequently to scientific publications. Information will also be
released to health departments and policy makers throughout the Great Lakes
basin, government agencies and through the scientific network as outlined in the
original proposal.
GREAT LAKES FISH AS A SOURCE OF MATERNAL AND FETAL
EXPOSURE TO CHLORINATED HYDROCARBONS
The Great Lakes have been monitored for contaminants in the water, the flora,
and the fauna. Several categories of pollutants, such as the polychlorinated
biphenyls (PCBs), and dichlorodiphenyl trichloroethane (DDT) and
dichlorodiphenyl dichloroethylene (DDE), have been investigated exten-
sively. Contaminants present in Great Lakes fish at levels exceeding the Food
and Drug Administration (FDA) standards include PCBs, DDT, and dieldrin.
These compounds are found in lake trout, perch, carp, salmon, and other
salmonoids, all of which are caught by sports anglers and their families.
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Dieldrin, PCB congeners, and DDT and its metabolite, DDE, are chosen as the
focus of this ATSDR-supported research project due to their persistence in the
environment and toxicological history in mammals.
The principal objectives of this research project are listed below:
1. Determine if the consumption of fish from Lake Michigan and its
surrounding tributaries in a poor, urban setting leads to toxic insults
to the developing fetus.
2. Collect biological samples from women and neonates to analyze
targeted chlorinated hydrocarbons known to bioaccumulate or
biomagnify via the aquatic food chain of Great Lakes fish.
3. Evaluate meconium as a biological specimen to predict infant in
utero exposure to toxic insult.
4. Establish a repository of biological specimens for later analysis of
newly identified toxic chemicals in the Great Lakes.
Previous studies of PCB exposure have demonstrated the correlation between
the ingestion of Lake Michigan fish and chlorinated hydrocarbon burdens.
This and other similar studies have focused on white, middle class populations
primarily in small towns and cities along the shores of the Great Lakes. Some
African Americans were included in the studies, but the numbers were not
large enough to provide adequate data on this racial group. However, there are
large populations that attempt to provide additional nutrition to their diets
through the consumption of locally caught fish. Populations living along the
lake shore of Chicago have ready access to Lake Michigan along the
breakwaters, and the harbors, and in the park lagoons adjacent to Lake
Michigan. The urban poor are a group with an increased need for good
nutrition and sources of protein, particularly during pregnancy. The relatively
poor nutritional status and poor prenatal care in the urban poor population puts
the developing fetus in an extremely high risk situation. The exposure of this
fetus to even low levels of potential toxins may have devastating effects. In
addition, the possibility of compromised chlorinated hydrocarbon metabo-
lism in African American populations raises concern for this group following
exposure (Hayes 1982; Kutz 1983).
Pregnant women who come to the University of Chicago Lying-in Hospital
for delivery will be interviewed and women identified as consumers of Great
Lakes fish will be invited to participate in the study. A matched control group
will be recruited in the same manner. Biological specimens, including
maternal blood, cord blood, placenta, adipose tissues (if Caesarean), infant
meconium, and breast milk, will be obtained from the fish eating group and
the control group of pregnant women (approximately 75% African American)
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residing along the shores of Lake Michigan in the southeastern area of
Chicago. Dieldrin, 9-hydroxy-dieldrin, DDE, and 9 PCB congeners (PCB 20,
28,52,101,118,138,151,153,and 180) will be analyzed by gas chromatog-
raphy-electron capture detection. A maternal health index and neonatal health
index will be obtained for each subject included in the study. Statistical
analysis of the tissue-toxin levels will be conducted between fisheaters and
non-fisheaters.
This research project is being conducted by Drs. Donald Waller, Ian Tebbet,
H.R. Al-Khalidi and Ms. Celeste Presperin (University of Illinois at Chicago);
in collaboration with Drs. Hans van der Van, Katrin van der Van, and Judith
Hibbard (University of Chicago).
PROGRESS AND PRELIMINARY RESULTS
Identify a subject population of pregnant African American women in an
urban Chicago area adjacent to Lake Michigan who regularly consume fish
obtained from local waters.
The identification of a subject population of pregnant African American
women has been proceeding at the University of Chicago Lying-in Hospital.
Interviews began during the middle of January 1993. The primary site for the
initial contact with patients was the prenatal care ward. These types of patients
were targeted early in the program due to their history of reasonable follow-
up and assurance of delivery at the University of Chicago Hospital. The
identification of subjects early in the prenatal care period allows greater
flexibility in obtaining a completed questionnaire and follow-up of any
questionable data.
In some cases during the inital interview, the test subject was not able to
identify the species offish or the location where it was caught. The person has
the opportunity, however, to contact someone and obtain the required infor-
mation before a follow-up interview during a later prenatal visit. The second
contact with the subject during the prenatal care greatly increases the ability
to complete the follow-up on these data not readily available during the first
interview.
Approximately 175 patients per month were interviewed during the period of
January 15 to April 30,1993. A total of 11 fisheating patients and 11 controls
were identified. This rate of recruitment would result in an additional 13
subjects before the end of the grant period (total 24 fisheaters). Several steps
have been taken to increase the number of subjects being interviewed and to
target populations that may provide additional subjects:
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An additional staff member will be hired who can contact patients
during off hours and weekends in the delivery room. This will
provide access to patients who do not participate in prenatal care at
the hospital and will maximize the number of patients that can be
interviewed.
2. Three additional clinical sites were investigated for participation
in this study. The Lakewood Clinic, affiliated with the Univer-
sity of Chicago Lying-in Hospital, is a maternal-child clinic primarily
serving the African American community.
A second community group located on the south side of
Chicago is associated with a large, low income African
American community called Altgeld Gardens. This study
population is currently the subject of a Centers for Disease
Control and Prevention (CDC) study because of the high
levels of pollution in the area. The community is located
adjacent to Lake Michigan and has a large lake (Wolf Lake)
that is used as a primary fishing site. There are direct water
links between Wolf Lake and Lake Michigan. This commu-
nity group has enthusiastically endorsed participation in the
study and has volunteered to perform the initial interviews to
identify potential pregnant fish-eaters. The identified subjects
will be formally interviewed for identification and formal
follow-up by the nurse coordinator.
The University of Illinois Hospital OB-GYN Clinic was
approached. This clinic serves a diverse community,
including a heavy concentration of African American women.
The location is close to the lake, but not as close as the other
sites. Because of the large number of public assistance
patients who come to the hospital from all over the southside
of the city, however, the possibility of identifying additional
subjects from this clinic is good.
3. A direct interview of people fishing along the lake shore will also be
attempted. The spring and summer seasons present an opportunity to
make direct contacts with individuals fishing along the lake shore
to identify pregnant consumers of Lake Michigan fish. Appropri-
ate alterations in our current IRB submission are underway to obtain
approval and include this mode of subject identification.
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The number of subjects required for statistical analysis is approximately 80
subjects in the fish-eating group and 120 in the control group. It is anticipated
that an adequate number of subjects will be identified for initial statistical
analysis before the end of the first grant period.
Obtaining biological specimens: maternal blood, cord blood, placenta, adi-
pose tissue (ifCaesarean), infant meconium, and breast milk from a control and
fish-consuming group
The specimens from the subjects are being collected by house staff at the
clinical site. The required specimen containers and labels for each patient
collection are prepared as kits and are available at the delivery room for
convenience and accuracy of collection. The sites for specimen containers and
temporary specimen storage were identified at the University of Chicago. Most
of the subjects have projected delivery dates due to their identification during
prenatal care. Personnel are alerted to the possibility of these patients beginning
labor, and preparations are usually made before their arrival at the hospital.
Specimens are hand delivered from the University of Chicago Lying-in
Hospital to the University of Illinois site on a regular basis. A central repository
for the specimens has been established at the University of Illinois. Split
samples of serum, placenta, and meconium were collected for analysis of lead
and mercury in May 1993. The samples will be kept frozen in appropriate
plastic culture tubes. Analysis of the metals will begin during the next grant
period.
Analysis of biologic specimens for selected PCB's, DDE/DDT and dieldrin
The team of investigators has been working to establish standardized proce-
dures for PCBs, dieldrin and DDT/DDE. The increased sensitivity of a new
gas chromatograph instrument will allow analysis of PCBs not originally
included in the original grant submission. The establishment of the analytical
procedures will include as many PCBs congeners as possible, with a focus on
those with established biological concern. A final list was submitted to ATSDR
on establishment of the analytical procedures for comparison to other labora-
tories performing PCB analysis. The laboratory contacted other participating
laboratories to determine which PCB congeners other ATSDR-investigators
are measuring for inclusion in their analysis.
SUMMARY OF PARTICPANTS' EVALUATION OF THE PROGRAM
The overall goal of the Great Lakes program is one of great importance. The
investigators are aware of the concern at all levels to identify potential sources
of chemical contamination from the environment. The currently funded
projects will increase the base information available to make future decisions
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on the recovery or the decline of the health of the Great Lakes and its impact on
those who use the lakes. The establishment of accurate and well controlled data
on the levels of various contaminants that enter the human food chain will be
critical to the success or failure of present and, especially, future efforts to
minimize or eliminate pollution in the Great Lakes.
/
The harmonization of the analytical procedures is an effort that should be
encouraged and supported. This type of effort will develop data that can be
compared among laboratories, among subject populations, and to data gener-
ated by future studies. It is vital to understand the differences and the similarities
of the various subject cohorts from the different geographical areas of study,
both now and in the future.
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CONTRIBUTION OF NURSING TO BEHAVIORAL CHANGES IN
OFFSPRING OF MOTHERS WHO CONSUMED LAKE ONTARIO
FISH: TWO METHODOLOGICAL APPROACHES
Some humans are ingesting large quantities of contaminated Great Lakes
fish, (e.g., from Lake Ontario), despite advisories issued by the New York
State Department of Health. These advisories are based primarily on
estimated reproductive and cancer risks. Although research has studied the
effects of individual toxic chemicals on human and animal behavior, very
little is known about the behavioral effects resulting from consumption of
mixtures of chemicals found in Great Lakes fish (e.g., polychlorinated
biphenyls [PCBs], dioxin, hexachlorobenzene (HCB),
dichlorodiphenyldichloro-ethylene (DDE), arid mercury). For example, no
data are available to determine whether behavioral differences exist between
adults who eat fish from the Great Lakes and those who do not or whether
some fisheaters are at greater risk than others. In addition, only one previous
study (Jacobson et. al 1984,1988) has been completed that addresses whether
babies whose mothers have eaten Great Lakes fish exhibit behavioral
changes. The Jacobson's study reported positive correlations between Lake
Michigan fish consumption by the mothers and maternal serum PCB levels,
which in turn, predicted cord PCB levels. The present research project
extends this groundbreaking work.
The following list highlights the research objectives of this ATSDR-sup-
ported multidisciplinary project:
1. Assess behaviors of babies born in Oswego, New York (on the
southeastern end of Lake Ontario), between October 1991 and
September 1994 to mothers who have or have not consumed Lake
Ontario fish. The researchers hope to identify a subset of behavioral
tests that are best at detecting behavioral changes due to the toxic
chemicals in Lake Ontario fish.
Pregnant women part way through their pregnancies are
interviewed at the only obstetrics/gynecology office in Os-
wego County. All women are re-interviewed after they have
given birth at the Oswego Hospital. The newborn babies are
tested (e.g., Brazelton's Neonatal Behavioral Assessment
Scale, Ballard, Apgar, in addition to physical measures).
Women who have eaten medium to high levels of Lake
Ontario fish, and an equal number who have never eaten
Lake Ontario fish, are asked to remain in the study and
become part of the longitudinal sample. Their children are
re-tested at 3, 6, 12,24, and 36 months (e.g., Pagan Test of
Infant Intelligence, Bayley, Early Language Milestones,
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Mobile Reward Shift Frustration test), and the women are
tested on three of the Neurobehavioral Evaluation System
(NES2) tests.
The researchers are also measuring more than 80 control
variables (e.g., social-economic status, mother's IQ, alco-
hol consumption), and will use them to identify childbearing
women whose children are at greatest risk.
2. Collect biological samples from women and neonates to analyze for
Great Lakes contaminants. Cord serum is being tested for 67 PCS
congeners, HCB, mirex, DDE, and lead. Breast milk samples
collected about two weeks after birth and again at 3 months are also
being analyzed for the same chemicals as cord serum. The mother's
hair will be tested for methylmercury; a piece of placental tissue will
be tested for responses associated with dioxin (P-450), and the
researchers are tissue-banking an additional piece of placenta for
future analysis.
3. Conduct neurobehavioral tests in animal models. To increase con-
trol over exposure levels, the researchers are also using experimental
methods with laboratory rats. Adult rats were fed a 30 percent diet
of Lake Ontario salmon for 20 days and tested in 7 different
behavioral tests. When "life is pleasant", there were no behavioral
differences, but when "life is made unpleasant," the rats fed Lake
Ontario salmon were much more reactive to negative events than
control rats fed Pacific Ocean salmon or no salmon. The researchers
also have shown that offspring of rat mothers fed Lake Ontario
salmon during gestation and for the first 7 days of nursing are also
more reactive to negative events.
4. Determine the effect(s) of paternal exposure to Great Lakes contami-
nants on their offspring. The researchers are currently testing the
effects of the mothers and/or fathers exposure to the toxic chemicals
prior to gestation, as well as the influence of exposure to the toxic
chemicals via nursing. By cross-fostering rat pups at birth, the
researchers can expose pups to the toxic chemicals in Lake Ontario
salmon during gestation and/or during nursing. If the same pattern
of results is found with humans as was determined with rats, then one
has a firmer basis for concluding that the correlational evidence with
humans is based on a cause and effect relationship.
This four-pronged approach, designed to determine the types of behavioral
changes in adults and offspring of humans and laboratory rats who have eaten
Lake Ontario fish, is being conducted at the Center for Neurobehavioral
Effects of Environmental Toxins and Department of Psychology at the State
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University of New York at Oswego by Drs. Helen Daly, Edward Lonky,
Jacqueline Reihman, Thomas Darvill, and David Sargent, in collaboration
with Drs. Brian Bush and Patrick Parsons (State University of New York at
Albany); Drs. Thomas Clarkson and Elsa Cernichiari (University of Roches-
ter Medical School); and Dr. James Olson (University of Buffalo).
PROGRESS AND PRELIMINARY RESULTS
All women planning to deliver babies in the Oswego Hospital (the only
hospital in the region) were invited to participate in this study. Those who
give informed consent complete an extensive interview covering fish con-
sumption, demographic, and control variable information. Women who have
eaten medium to high amounts of Lake Ontario fish and an equal number of
women who have not eaten any Lake Ontario fish are asked to become part
of the longitudinal sample. Their babies are tested at several points in time.
The babies of the remaining mothers are tested only through hospital
discharge, non-longitudinal sample.
Daily Activities of Research Program on a Weekly Basis 20-week Interview
The Oswego OB-GYN practice is very supportive of this study. The nurse
practitioner introduces the study to the woman during her first visit and gives
her a copy of the pamphlet describing the study. At the mothers scheduled
sonogram visits, the interviewer interviews all mothers who agree to partici-
pate (approximately 40 mothers a month). Each interview takes between 20
and 45 minutes. Data are brought back to the main office and checked, and
more than 600 pieces of data are entered into the computer for each interview.
Selection of Longitudinal Sample
Fish-consumption questionnaires are scored, and total lifetime fish consump-
tion is converted into total equivalent PCB pounds (the grantee has rated each
fish species for amount of PCBs based on data provided by the New York
State Department of Environmental Conservation). These data are then used
to determine whether the fish-consumption meets the criterion (> 40 PCB-
equivalent pounds; value suggested by Dr. Jacobson) for inclusion in the
longitudinal sample. At the beginning of each month, the number of
fisheaters with expected due dates 60 to 90 days from then are determined.
The research team then randomly selects an equal number of non-fisheaters
from the pool of subjects with expected due dates within the same 30-day
period. The team creates the lists needed to inform the hospital staff,
behavioral assessment team, interview team, and person shipping blood
samples, of those mothers who are in the longitudinal and non-longitudinal
sample.
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Birth
The research team calls the hospital every morning at 7:30 a.m. ( 7 days a
week) to obtain information about which babies were born. The assigned
person on the behavioral assessment team goes to the hospital and adminis-
ters the Ballard and Brazelton tests. Feedback is given to the mother, the
Obstetrics Complications Scale is completed, and informed consent for
continued participation in the study is secured (approximately 40 babies a
month). All data are returned to the office and, checked, and more than 320
data points are entered. Since the Brazelton is administered twice (between
12 and 18 hours of age, and again between 24 and 36 hours), very often several
staff members will go to the hospital.
The hospital interviewer is also informed about which babies were born, goes
to the hospital to administer the follow-up interview, and obtains a hair
sample for mercury analysis. All data are returned to the office and, checked,
and more than 200 data points from the interview are entered.
Blood, Hair, and Breast Milk Samples
The behavioral assessment team brings all samples from the hospital.
Samples are coded and sent to the laboratory for analysis of lead, total PCBs,
and 69 PCB congeners, mirex, DDE, HCB, and mercury. The investigators
anticipate being able to determine which combination of these toxic chemi-
cals correlate most highly with both fish consumption and behavioral changes
in both the babies and their parents. The results of the chemical analyses are
entered into the computer as soon as the data arrive. Both the obstetrician and
the pediatrician are alerted if the lead level is elevated.
If mothers are breast feeding, then arrangements are made to pick up samples
the first and second week after each birth and at 3 months. The milk is frozen
and then sent to Dr. Bush for analysis (69 PCB congeners, mirex, DDE, HCB,
and a total PCB value). Dr. Bush's laboratory has completed analysis of 29
milk samples as of May 1993.
3-Month Testing Procedures
When a baby is 2 months old, a letter is sent to remind the mother of the 3-
month (53-week gestational age) visit. One week later, a call is made
schedule the appointment, and a postcard is sent to remind the mother of the
visit. Three-month testing involves home visits on 3 consecutive days: Day
1 (about 1.5 hours) includes the Fagen Infant Test of Intelligence, IBQ, ELM,
Feelings Scale, and Mobile Acquisition phase; Day 2 (about 1 hour) includes
the Bayley and Mobile Acquisition; Day 3 (about .5 hours) includes the
Mobile Shift phase. The research team averages about 3 babies a week (9
home visits), although some weeks are much higher (5 babies with 15 home
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scored, and data (more than 275 points per baby) are checked and entered into
the computer.
6-Month Testing Procedures
One month before the baby is 6 months old (67 to 69-week gestational age),
a letter is sent to remind the mother of the 6-month visit. One week later, the
investigator will call to schedule the appointment. Six-month testing
involves one 2-hour testing period: Pagan Infant Test of Intelligence, IBQ,
ELM, Feelings Scale, Hassles Scale, Bayley, and HOME. All protocols are
scored, and data (more than 300 points per baby) are checked and entered into
the computer. Testing began on April 26,1992. (Note: these studies were
begun prior to ATSDR funding)
1-Year Testing Procedures
The first baby reached a gestational age of 1 year on October 2,1992. Contact
with the mother is made as previously stated for other testing. Testing of the
baby and the mother takes 2.5 hours: Pagan Infant Test of Intelligence, IBQ,
ELM, Feelings Scale, Hassles Scale, and Bayley. The team also tests one
parent (typically the mother) on three NES scales (symbol-digit substitution,
color-word, and serial digit learning) and the Peabody test.
Part II: Effects of Nursing Using the Animal Model
Breeding and Behavioral Testing
Breeding and cross-fostering for the nursing study has been very successful;
the study will be completed in the first year. Fifty-four female rats were bred,
and 18 were fed a 30 percent diet of Lake Ontario salmon (LAKE), 18 a 30
percent diet of control Pacific Ocean salmon (OCEAN), and 18 a control
mash diet (MASH). The percentage of successful births was high (Mean =
96%; 94.4% in the LAKE and OCEAN groups and 100% in the MASH
group). Litters born within 24 hours of each other were successfully cross-
fostered (LAKE to LAKE; LAKE to OCEAN, OCEAN to LAKE, OCEAN
to OCEAN, and an additional MASH to MASH control). Numerous
offspring are in each condition. As predicted (based on our previous work),
there were no physical differences (e.g., success of litters, size of litters, sex
ratio, weight of pups, righting reflex) among the groups. The offspring are
now adults, and behavioral testing has been initiated on the Progressive Ratio
task and the Depression Effect task.
Additional Research
(1) The investigators are testing the effects of a Lake Ontario fish diet on
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behavior on a Fixed Ratio Schedule and a DRL schedule (half the
subjects have been tested).
(2) The investigators have completed analysis of the two studies (pro-
gressive ratio and depression effect) that show that if only the
grandmother is fed Lake Ontario salmon, her grandsons show behav-
ioral changes. The researchers have begun writing the manuscrip for
publication.
HEALTH RISKS FROM CONSUMPTION OF GREAT LAKES FISH
This research program is the first step in a long-term collaborative effort by
a consortium of institutions involved in epidemiologic investigations of
Great Lakes toxicants. One important component of this program is to recruit
and maintain subpopulations of individuals in Michigan who have relatively
high exposures to Great Lakes fish, as well as companion groups having
relatively lower exposures. This research program will extend and enhance
the valuable cohorts exposed to polyhalogenated biphenyls previously
recruited and maintained by Dr. Harold Humphrey at the Michigan Depart-
ment of Public Health.
The focus of this research program is to recruit young women of childbearing
age who agree to participate in epidemiology studies and to examine the effect
of fish consumption and related factors on human reproductive outcomes.
Measurements will be made using fluids and tissues from newly recruited
individuals to assess exposure to specific toxic chemicals in Great Lakes fish,
and questionnaires will be used to obtain information on diet, lifestyle, and
demographic status.
This collaborative research program consists of two research projects:
Project 1- The Establishment of a Cohort of Persons of Reproductive Age
Exposed to Great Lakes Fish Contaminants: A Pilot Project
The Great Lakes contaminants of primary interest in this project are the
polychlorinated biphenyls (PCBs) and methylmercury (Hg). The objectives
of this project are:
1. Establish a cohort of men and women of reproductive age who are not
using effective means of birth control and who represent a sufficient
variation in exposure to Great Lakes fish contaminants to allow for
the investigation of Great Lakes fish consumption, and potentially
associated infertility and early pregnancy loss.
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2. Characterize the cohort in terms of exposure to specific contaminants
in Great Lakes fish through survey instruments and measurements of
biological specimens.
Subjects for this pilot study were chosen from the Department of Natural
Resources (DNR) list of licensed anglers in selected counties of Michigan for
the year 1992. These counties were selected from the 11 counties that served
as the source of subjects for the Michigan Fish-eaters Cohort, initiated and
maintained since 1980 by the Michigan Department of Public Health.
Studies of the anglers in the Michigan Fisheaters Cohort indicate that these
sport fishermen and their families consumed three times more fish than the
national average. In addition, the median serum PCB levels reported in the
anglers (n=572) was 21.4 ppb, compared to a median of 6.6 ppb in the control
group (n=419) (Humphrey 1988a). It seems likely therefore, that the cohort
of anglers (and their spouses or partners) that will be identified in this study
will have a high probability of exposure to Great Lakes fish contaminants.
A total of 500 subjects comprising 250 anglers and their spouses/partners
were selected for this study. To assess the role of maternal fish consumption
among anglers themselves, sampling is being done to assure that one-third
(n=83) of the anglers are women.
Surveys are being administered to determine historical and current Great
Lakes fish consumption habits of these men and women, as well as other
possible sources of exposure to PCBs and mercury. Analyses for total PCBs,
nine pesticides (i.e., DDT, DDE, HCB, beta-BHX, oxychlordane, heptachlor
epoxide, trans-nonchlor, dieldrin, and mirex), lead, and mercury are being
performed. A protocol for the prospective evaluation of paternal exposures
prior to and during their partners pregnancies is being developed.
Project 2- Variations of Mercury Body Burdens During Pregnancy
The objective of this project is to determine whether mercury body burdens,
as reflected in concentrations of mercury in hair and blood, are stable in
pregnant and nonpregnant women who have differing exposures to methyl-
mercury from consumption of freshwater fish. Information gained from the
study will provide currently unavailable information about the changes in
mercury body burdens during the course of pregnancy and, thus, the potential
for changes in fetal exposures during gestation.
In addition, results obtained from nonpregnant women will be compared to
those from pregnant women, thereby providing information on the effect of
pregnancy on the stability of body burdens of mercury. Sequential hair
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analysis will provide a chronological record of mercury exposure during a 9-
month period. A sufficient number of blood mercury analyses will be
conducted to determine whether the blood to hair ratio is constant in pregnant
and nonpregnant woman.
This research is being conducted by Drs. Lawrence Fischer, Michael Kamrin,
Emmett Braselton, Jeanne M. Courval, and Nigel Paneth and Ms. Jane
DeHoog and Ms. Kay Trosko (Michigan State University) in collaboration
with Dr. Harold Humphrey (Michigan Department of Public Health) and Dr.
Anne Sweeney (University of Texas at Houston).
PROGRESS AND PRELIMINARY RESULTS
Identify a population of men and women of reproductive age who have a high
probability of Great Lakes fish consumption, based on:
a) having been registered as a licensed angler by the state of Michigan
in 1991, and/or
b) being the spouse/partner of a licensed angler in Michigan during
1991.
A list of licensed anglers has been obtained from the Michigan Department
of Natural Resources (DNR) for the 1991 fishing season. These licensed
anglers are from 16 Michigan counties which are listed by the DNR as having
fishing areas of concern.
Originally, the investigators proposed to select individuals from the 11
counties sampled by the Michigan Department of Public Health (MDPH) for
the Michigan Fisheaters Cohort (Humphrey 1976). However, because these
11 counties were in the southwest portion of the State along the Lake
Michigan coast, the investigators felt that assessing anglers residing only in
these counties may not provide a representative sample of all young licensed
anglers in Michigan. Using the 1991 Michigan Fishing Guide published by
the DNR, the investigators selected counties that have fishing areas of
concern, that are located along each of the Great Lakes touching the State of
Michigan (i.e.,Lakes Michigan, Superior, Huron, and Erie).
Establish a cohort, derived from this exposed population, who are "at risk"
of pregnancy (fathering a child, if male, and conceiving a child, if female}.
as ascertained through a survey instrument
In the original proposal, the investigators planned to send letters of introduc-
tion to prospective cohort candidates explaining the project and keying them
to expect a telephone call from a project employee. The telephone call would
explain the project in more detail and give the contacted individual an
opportunity to ask questions. The caller would conclude by asking questions
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to assess if the individual met the necessary criteria, (i.e., planning to conceive
a child in the next five years, for their inclusion into the project.
The investigators have revised this protocol to be more timely and cost
effective. They have combined the introductory letter with two short question-
naires; one for a male and one for a female, which replace the expensive
telephone call. Each letter with two questionnaires also includes a self-
addressed envelope to return the questionnaires. This first mailing will be
followed up 10 mailing days later with a postcard prompt. If no response is
obtained from an individual, a second mailing will be sent, similar to the first,
but will include a new explanatory letter. The mailing screen is being
undertaken by county in a step-wise fashion until 500 candidates are enrolled.
An 800 telephone number was set up to enable potential participants to call for
answers to any questions about the project.
Characterize the cohort in terms of exposure to specific contaminants in Great
Lakes fish through survey instruments and measurements of biological speci-
mens
The fish consumption survey has been revised based on suggestions by the
Dr. John Vena, project leader at State University of New York in Buffalo,
another ATSDR investigator. The following revisions were made:
0 The investigators expanded the lower end of fish consumption num-
bers in the "Average Number of Fish Meals" tables to be more
specific in the number of meals eaten by infrequent fish-eaters. The
upper estimate of number of fish meals eaten was combined into one
category. It was felt that doing this would more accurately reflect
the fish consumption of the majority of the population.
0 Bass was added to all tables asking about the frequency of eating
specific species of fish.
0 The investigators included a question asking for number of meals of
sport fish eaten that were caught from other waters in Michigan (i.e.,
inland lakes), not specifically listed in the tables.
0 The investigators included a question asking for number of meals of
sport fish eaten that were caught from waters outside Michigan.
0 The investigators eliminated some of the knowledge of risk in eating
Great Lakes fish questions, as well as some of the risk-taking
behavior questions.
0 Questions about health status, health behavior, reproductive informa-
tion, and risk-taking behavior will become part of the accompanying
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baseline questionnaire (specific questionnaires for males and for
females).
The 1991 Michigan Fishing Guide (DNR 1991) was used to establish
Great Lake areas of fishing concern and Great Lake tributaries of
fishing concern specific to Michigan to replace New York waters of
fishing concern. Specific species of fish caught in those waters that
carried consumption warnings were used to replace species impor-
tant to New York waters.
Tracking System for Recruitment
The data manager for the project, developed a computerized tracking system for
monitoring the progress of recruitment efforts. This program accounts for each
individual (licensed angler) to whom a letter was sent. Each potential
participant is defined in terms of participation ID code, status, partner informa-
tion, current pregnancy status, and future pregnancy plans. If an individual
refuses to participate, either by not responding to all three contact attempts, or
by notifying us that they do not wish to enroll, he or she will be designated as
a refusal. If the licensed angler responds to the survey, but does not meet all
the eligibility criteria above, he or she will not be contacted further.
Timetable
The identification of subjects, assessment of their eligibility and willingness to
participate, is expected to take 18 months. Data collection, survey, and
specimen collection is expected to take an additional six months. Refining and
analyzing the data, plus writing and publishing the manuscripts, are expected
to take 12 months. Approximate dates of completion are:
10/92 to 4/94 Identifying subjects, screening for eligibility and
willingness to participate
5/94 to 11/94 Surveying, collecting data, and specimen collect-
ion
11/94 to 10/95 Refining and analyzing the data. Writing and
publishing results.
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Project 2. Variations of Mercury Body Burdens During Preganncy
The objective of this project is to determine whether mercury body burdens, as
reflected in concentrations of mercury in hair and blood, are stable in pregnant
and nonpregnant women who have differing exposures to methylmercury from
consumption of freshwater fish. Information gained from the study will
provide currently unavailable information about the changes in mercury body
burdens during the course of pregnancy and thus the potential for changes in
fetal exposures during gestation.
In addition, results obtained from nonpregnant women will be compared to
those from pregnant women, thereby providing information on the effect of
pregnancy on the stability of body burdens of mercury. Sequential hair analysis
will provide a chronological record of mercury exposure during a 9-month
period. A sufficient number of blood mercury analyses will be conducted to
determine
whether the blood to hair ratio is constant in pregnant and nonpregnant woman.
Recruitment of Cohort
During the initial period of the grant, recruiting of the cohort was in progress.
Women are contacted through a questionnaire and their suitability for partici-
pation in this study is evaluated.
Mercury Analysis
The first 6 months of the grant were devoted to initiating the analytical method
for mercury in hair. Instrumentation has been purchased, a research assistant
hired, and a laboratory for mercury analysis established as of May 1993. The
new analytical equipment is being evaluated for its reproducibility to measure
extremely low levels of mercury in standard reference samples.
Initial efforts have been directed toward (1) familiarization of technical staff
with instrumental operating parameters; (2) determination of linear operating
ranges (parts per trillion, ppt to parts per billion, ppb) at low and high sensitivity
settings of the instrument; and (3) verification of accuracy using National
Institute of Standards and Technology (NIST) reference materials.
The instruments can be operated in two ranges of sensitivity, depending on the
anticipated sample size and sample Hg concentrations. The low sensitivity
range is accomplished with a 1 ml sample loop, and small volumes of reagents
for Hg vapor generation. The linear range is approximately 100 ppt to 10 ppb
(in sample introduced). The high sensitivity range is from 0 to 100 ppt, using
a 5 ml sample loop, and larger volumes of reagents. Surprisingly, the
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investigators obtained much better precision working with this range, and have
focused their efforts on optimizing these conditions. It appears that the larger
volumes allow better overall precision. The investigators have verified the
accuracy of the method using NIST reference material, as shown in the
following table. Analytical development work is continuing toward a reproduc-
ible method for analysis of mercury in small amounts of hair.
LOOP
NIST REFERENCE
MATERIAL
CERTIFIED VALUE
ppt
RANGE
FOUND
rep. 1 rep. 2
5ml
5ml
1ml
Oyster Tissue SRM
1566a
Bovine Liver SRM
1577b
Albacore Tuna RM 50
64,200
( 57,500-70,900)
3,000
(non-certified)
950,000
(1,050,000-850,000)
62,100 59,800
2,650 3,060
938,000
Analytical development work is continuing toward a reproducible method for analysis of mercury
in small amounts of hair.
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CONSORTIUM FOR THE HEALTH ASSESSMENT OF GREAT
LAKES SPORT FISH CONSUMPTION
This research project consists of an integrated program of epidemiology,
survey, and laboratory studies to investigate the human health impacts of
Great Lakes sport fish consumption. It is a consortium of five State health
departments (Illinois, Indiana, Michigan, Ohio, and Wisconsin); the Univer-
sity of Wisconsin (State Laboratory of Hygiene, Department of Preventive
Medicine); and the Wisconsin Department of Natural Resources. The
objectives of this research program include the following:
1. Epidemiology
(a)
(b)
(c)
(d)
2. Survey
(a)
(b)
(c)
3. Laboratory
Identify a cohort of 3,000 to 5,000 frequent Great Lakes
sport fish consumers (e.g., Great Lakes charter boat captains
and anglers and their families).
Analyze approximately 655 blood samples to determine the
array of mixtures of bioconcentratable organic pollutants
frequent consumers of sport fish may be exposed to and
evaluate which substances may serve as good markers for
total exposure.
Assess the impact of past and current dietary practices on
current mixtures of contaminant body burdens.
Estimate the disease risk (adverse reproductive outcomes,
low birth weight infants) attributable to ingestion of con-
taminants in sport fish.
Characterize the contribution of sport caught fish to the diet
of Great Lakes State residents.
Describe the demographic characteristics of frequent Great
Lakes fish consumers and non-consumers.
Estimate the fish consumption potential for impact on the
public's health state-wide and basin-wide.
(a) Develop and evaluate new analytic methods to characterize
an array of Great Lakes toxic contaminants in human blood
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samples using gas chromatograph/mass spectrophotometry
techniques.
(b) Improve the detection of Great Lakes toxic substances in
body fluids, tissues, and other biologic matrices through
research and laboratory coordination, in partnership with the
Great Lakes Protection Fund.
4. Information Dissemination
(a) Develop and evaluate information dissemination and inter-
vention strategies anticipated to result in reducing human
bioaccumulation of sport fish contaminants.
(b) Increase public awareness of the health implications asso-
ciated with toxic pollution via the Great Lakes Advisory
Task Force.
5. Inter-State Surveillance Coordination
This activity will advocate to enhance the role of state and territorial
health departments in surveillance and prevention of environmentally
related morbidity and mortality, through coordinated efforts with ATSDR
and other Great Lakes States health and environmental departments.
Through the establishment of the consortium of Great Lakes state health
departments and the University of Wisconsin, this project integrates existing
public health disease surveillance and prevention expertise with university
epidemiologic and laboratory science resources to achieve the following
goals:
1. Significantly strengthen the scientific understanding of the relation-
ship among prolonged Great Lakes sport fish consumption, persis-
tent contaminant levels in human blood samples, and the potential
for adverse reproductive health impacts.
2. Through collaboration of consortium members and other grantees,
standardization of protocols to characterize sport fish consumption
and reproductive health which will allow comparisons between
already existing data and ongoing research.
3. Demonstrate the feasibility of establishing interstate access and data
confidentiality agreements to assure full utilization of existing state
maintained health data systems, which will allow interstate data
sharing for the continuing characterization of the surveillance co-
horts.
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This ATSDR-supported research program is being conducted by Drs. Henry
Anderson, Lawrence Hanrahan, and Ms. Claire Falk (Wisconsin Department
of Health and Social Services); in collaboration with Drs. William Sonzogni
and Marty Kanarek (University of Wisconsin), Mr. Jim Amrhein (Wisconsin
Department of Natural Resources); Dr. Kim M ortenson (Ohio Department of
Health), Dr. Greg Steele (Indiana Board of Health); Dr. Tom Long (Illinois
Department of Public Health); and Dr. Harold Humphrey (Michigan Depart-
ment of Public Health).
PROGRESS AND PRELIMINARY RESULTS
Establish a cohort of frequent sport fish-consumers
The cohort consists of licensed charter boat captains who obtained a license
in one of the following five states — Illinois, Indiana, Ohio, Michigan, or
Wisconsin. The cohort also includes Wisconsin anglers who have been
previously studied.
Charter captains must obtain a license in each state, that is valid for one year.
Each State maintains a database of all the licensed captains. Mailing lists
containing each captain's name, address, and telephone number were gener-
ated at each of the five consortium states from these databases. Consortium
members reviewed the respective state records for completeness and obtained
missing variables. The lists from the five States were merged and identifi-
cation numbers assigned to each captain. Approximately 2,500 captains and
their spouses are being surveyed.
The Wisconsin anglers were obtained from the list of subjects who partici-
pated in past studies (Fiore 1986; Sonzogni et al. 1991). A copy of the file
maintained by the researchers was used to generate each angler's name,
address, and telephone number; 175 anglers and their spouses are being
surveyed.
Ascertain the cohort's demographic and Great Lakes fish consumption
patterns
Information is being obtained from the cohort members via of a computer-
assisted telephone interview. The following discussion describes the survey
design and explains the five modules.
(a) Charter boat captain and Wisconsin angler survey design
Core consortium members held a number of design meetings to construct the
survey to ensure a high degree of validity and reliability. Researchers from
the CDC also reviewed the survey and gave their expert advice.
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The survey is being conducted by the Wisconsin Survey Research Laboratory
(WSRL). Two pretests were conducted, the first on a sample of female anglers
(n = 15); it was necessary to assess the sensitivity of questions in the
reproductive module. The second pretest was on a random sample of the charter
boat captains and their spouses (n = 15). All modules were tested. During the
pretests, researchers from the core consortium group listened to the interviewer
and respondent to assess the delivery and the reactions to the questions.
(b) Description of the survey modules
To ascertain demographic and fish consumption patterns from the cohort,
WSRL is conducting a telephone survey comprising five modules
l.Fish
2. Fish summary
3. Demographic
4. Male reproductive module-other children
5. Female reproductive
The Fish Module contains questions about the subjects' sport fish and fish
consumption habits. The survey is designed to obtain weekly or monthly fish
consumption habits of the respondents during the last 12 months and in the last
week. The species listed are lake trout, brown trout, rainbow trout, chinook,
coho salmon,carp, catfish, perch, smelt, or walleye. There is also an open-ended
question for other species of the Great Lakes sport-caught fish. Both the cohort
member and his or her spouse are asked independently if he or she ate any fish
in the last 12 months, either purchased or sport caught. If the response is
positive, the respondent completes the module.
There are limitations in obtaining diet history data, one is the ability to recall
diet accurately. To minimize recall bias, data is being collected on fish
consumption habits in the last week from both the cohort members and their
spouses. The survey is designed to quantify the number of sport caught fish
meals consumed in the last week. An open ended question is included to allow
the respondent to list the fish species they have consumed and how each was
prepared (e.g., skinned, filleted, baked). Information about where the fish was
caught, a Great Lake (specific), stream, river, inland lake, pond, or ocean is also
being obtained in this module.
This module also includes questions about health advisory awareness. Respon-
dents are asked if they have ever heard of the advisory, and if they follow the
guidelines on fish species and size, fishing location, preparation of the fish, and
consumption frequency. If their sport fish or Great Lakes sport fish consump-
tion habits have changed over the years, questions will ascertain whether they
were due to the advisory.
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The Fish Summary Module focuses on the number of years cohort
members have been eating sport-caught and Great Lakes sport-caught fish.
Consumption habits over the years will also be determined. Respondents
are asked whether health advisories have influenced their decisions to
change their habits about species and/or quantity offish eaten. This module
also identifies who is most knowledgeable about the types of sport caught
fish consumed by the members of the household.
The Demographic Module ascertains the following characteristics on each
cohort member: date of birth, race, marital status, level of education, height,
weight, and total household income.
In The Male Reproductive Module, the category of other children was
added to the survey to obtain information about children the captain may
have had from any previous relationships. The name, birth date, and sex of
each child born since 1970 from another relationship are obtained. The
interviewer also asks the captains or anglers for the biological mother's or
father's name, address, and telephone number in case the researchers need
to contact her or him for additional information. Also mother's smoking
and alcohol habits during each pregnancy are obtained from the captain or
angler in this module.
Female spouses of the captains or anglers are asked questions in the
Reproductive Module. Information about each child the spouse has had
with the cohort member is obtained. Information includes the child's name,
date of birth, sex, and weight at birth. The spouse's fish consumption habits
during each pregnancy are collected. Information about other risk factors
that may be associated with adverse reproductive outcomes is also collected
about the spouse's lifestyle during each pregnancy, specifically, smoking
and alcohol use, prenatal care, and prescriptive and nonprescriptive drugs.
Examine the dietary and demographic profiles of Great Lakes toxicants (body
burdens) in a stratified cohort
Blood specimens were collected in the 2nd year of funding and analyzed for
several environmental contaminants. A pilot study is being conducted on 30
subjects, captains and their spouses, from three of the five states - -Michigan,
Ohio, and Wisconsin. Selection of the participants was based on their sport fish
consumption history. The captains reported they ate at least one fish meal a
week.
Pilot participants completed the survey before the remainder of the cohort was
interviewed. Blood and urine samples are being collected and will be tested for
the following analytes: congener specific polychlorinated biphenyls (PCBs),
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chlorinated hydrocarbon pesticides, PCDDs, PCDFs, and coplanarPCBs, mer-
cury, arsenic, and 12 nonpersistent pesticides. Laboratory analysis will be done
at the Centers for Disease Control and Prevention Laboratory. Results from this
analysis will determine laboratory testing for the full study.
When the blood and urine samples are collected, the pilot participants are asked
to complete a questionnaire. The exposure history questionnaire will obtain
information about contaminant exposures from sources other than consuming
contaminated sport fish. Consortium members in each State enter data into an
epidemiologic information (EPI INFO) file, and send it to Wisconsin via PC
Wonder. Information will then be uploaded to the Statistical Analysis System
(SAS) and merged with other study data.
COHORT REPRODUCTIVE OUTCOME STUDY
Ascertain the cohort'spopulation birth outcomes
The telephone survey, specifically the reproductive module, is designed to
obtain information about the captain's or angler's children. Some of the
information obtained from the reproductive module will be validated by
retrieving the birth records of all children born since 1970. Consortium
members staff have researched the storage methods for birth records in all five
States. The majority of the information is stored in computer files maintained
by vital records departments. This information will be uploaded into Statistical
Analysis System (SAS) for data analysis. Information that is not in the computer
will be manually entered into an EPI INFO file. These files will be uploaded
into SAS and merged with the other information.
Each consortium state receives a list containing pertinent information about
each child born in the respective State. The consortium members retrieve the
birth record data and send it via PC Wonder to the core consortium members.
Examine interrelationships between birth outcome, dietary (exposure) risks.
and demographics
The telephone survey is designed to obtain information about the sport caught
fish consumption habits and demographic variables of captains or anglers and
their spouses. In year 1, consortium staff members have made efforts to ensure
proper transfer of data from the WSRL to the Wisconsin Division of Health.
Meetings have been held between the Wisconsin staff and the staff from the
WSRL to discuss the layout of the SAS datasets and to design a coding scheme
to ensure proper tracking of all data collected. Analysis of the survey data will
occur in 2nd year of funding.
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NESTED COHORT-LOW BIRTH WEIGHT CASE-REFERENT
STUDY
The objective of the nested cohort, low birth weight case-referent study is to
ascertain the relationship between low birth weight (less than 2,500 grams),
Great Lakes toxics exposures, demographic, dietary, and other risk factors in
30 low birth weight cases and 30 matched controls. This objective was
achieved by selecting 30 low birth weight birth certificates and 30 matched
controls from the cohort reproductive outcome study. The demographic risk
factors of case and referent parents will be examined, and Great Lakes toxic
body burdens will be assessed for all parents.
BEHAVIORAL RISK FACTOR SURVEY - FOUR-STATE PILOT
SURVEY
The Behavioral Risk Factor Survey (BRFS) study has the following objec-
tives:
1. Pilot test a dietary fish consumption module in four Great Lakes
States
2. Assess the utility of BRFS to characterize sport fish consumption
Statewide.
3. Describe the demographic characteristics of frequent Great Lakes
fish consumers and non-consumers.
4. Estimate the fish consumption potential for impact on the public's
health in each state.
Sport fish consumption patterns and advisory awareness of the general
population in four of the Great Lake States will be characterized through the
addition of a fish consumption module to the monthly Behavioral Risk Factor
Survey conducted in each of four chosen Great Lakes States (WI, PA, MN,
MI). Approximately 4,917 adults will be questioned during the 1993 BRFS
year period.
Pilot test a dietary fish consumption module in four Great Lakes States
The fish consumption module was added to the BRFS in Wisconsin during
year 1, but was not added to the BRFS in Michigan, Minnesota and
Pennsylvania, although efforts were made by researchers to do so. The core
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consortium group organized several teleconferences with members of these
above States to provide information about the study and to request their
participation. Dr. Henry Anderson also presented information about the study
at the annual BRFS meeting in Atlanta, Georgia.
Assess the utility ofBRFS to characterize sport fish consumption Statewide
The fish consumption module was added to the BRFS in the state of Wisconsin
during year 1. In 1993,125 surveys were completed each month, a total of 1,168
were completed from January to September 30, 1993. Data analysis is
underway.
Describe the demographic characteristics of frequent Great Lakes fish consum-
ers and non-consumers
The BRFS has been completed on 1,168 households as of September 30, 1993.
The demographic characteristics collected from frequent fish consumers and
non-consumers is being analyzed.
POINT-IN-TIME SURVEYS - ALL EIGHT GREAT LAKES STATES
The point-in-time surveys have four objectives:
1. Implement a quarterly point-in-time sport fish consumption survey in
all eight Great Lakes States.
2. Describe the demographic characteristics of frequent Great Lakes fish
consumers and non-consumers.
3. Estimate the fish consumption potential for impact on the public's
health state-wide and basin-wide.
4. Characterize sport fish consumption patterns and advisory awareness
of the general population state-wide and basin-wide.
Four surveys of 2,000 samples each will contact a total of 8,000 households for
this investigation.
Implement a quarterly point-in-time sport fish consumption survey in all eight
Great Lakes States
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Three of the four point-in-time sport fish consumption surveys in all eight Great
Lakes States are complete. In 1993, the first quarterly survey started in mid-
April and was completed by May; 2,031 households were surveyed. The
second quarterly survey began in mid-July and was completed in August; 2,026
households were surveyed. The third quarterly survey began in mid-October
and ended in November; 2,168 households were surveyed. The final survey
began in January 1994. The data are being analyzed.
Describe the demographic characteristics of frequent Great Lakes fish con-
sumers and non-consumers
Three of the four point-in-time sport fish consumption surveys in all eight Great
Lakes States are complete. Results of the demographic characteristics of
frequent Great Lakes fish consumers and non-consumers were presented at the
ATSDR Great Lakes Human Health Effects Research Symposium.
Estimate the fish consumption potential for impact on the public's health state-
wide and basin-wide
Three of the four point-in-time sport fish consumption surveys in all eight Great
Lakes States are complete. Estimates of the fish consumption potential for
impact on the public's health state-wide and basin-wide were presented at the
ATSDR Great Lakes Human Health Effects Research Symposium.
Characterize sport fish consumption patterns and advisory awareness of the
general population state-wide and basin-wide
Three of the four point-in-time sport fish consumption surveys in all eight Great
Lakes States are complete. Data have beev analyzed and results were presented
at the ATSDR Great Lakes Human Health Effects Research Symposium.
LABORATORY ANALYSIS
Mixtures Method Development
This laboratory based investigation will develop and evaluate a new method to
characterize an array of Great Lakes toxic contaminants in human blood using
gas chromatograph/mass spectrophotometry techniques. Two spectra libraries
are being used to detect non-polar organic compounds in human blood
specimens: the Great Lakes high concern library (3 3 priority chemicals) and the
EPA/NIH/NBS spectra library.
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Considerable work has been done to develop a standard operating procedure for
the mixtures analysis. Some fish samples were analyzed as part of a pilot round
robin involving several major laboratories.
GREAT LAKES PROTECTION FUND CHALLENGE
In partnership with the Great Lakes Protection Fund, the objective of this
project is to carry out research and laboratory coordination that will improve the
detection of Great Lakes toxins in body fluids, tissue, and other biologic
matrices.
Funding for this part of the study (jointly funded by the Great Lakes Protection
Fund and ATSDR) was resolved in August 1993, so work is just beginning. The
following activities are ongoing:
o A workshop was be held to discuss methods of measuring Great Lakes
contaminants.
o A super critical fluid extractor has been purchased and set up.
o Research and method development has begun. A new super critical
fluid extractor on sediment and tissue is in the process of being tested
(comparing the results to traditional extraction methods).
INFORMATION DISSEMINATION
The consortium supports the development and evaluation of information
dissemination and intervention strategies to help reduce human bioaccumulation
of sport fish contaminants. This includes providing health information to the
subjects of the research project and their medical professionals. Through the
Great Lakes Fish Advisory Task Force, the consortium will increase public
awareness of the health implications from exposure to Great Lakes pollution.
Data analysis and a summary of the results are being sent to each subject. A
summary of the key findings of the Wisconsin Charter Boat Captain Study
conducted in 1991-1992 was sent to study subjects. All of the subjects were
Wisconsin, residents and many of them are participating in this study. The key
findings included a summary of the study results and a summary of poly chlo-
rinated biphenyl (PCB) levels in subjects who participated in studies conducted
by the WI Division of Health since 1986. The summary also made the subjects
aware of this new study. Informing the subjects of the previous study results
and notifying them of this new study, helped to increase awareness among the
charter boat captains and increase participation in this new study.
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The core consortium group drafted a fact sheet that is being sent to each subject
in the study. The fact sheet contains basic information about the study, including
who is conducting the study, who is funding the study, and the general protocol.
It also contains the name and telephone number of a consortium member from
each State who can be contacted for additional information.
INTER-STATE SURVEILLANCE COORDINATION
Through coordinated efforts with ATSDR and other health and environmental
departments in the Great Lakes States, the objective of this activity is to advocate
enhancement of the role of state and territorial health departments in surveil-
lance and prevention of environmentally related morbidity and mortality.
PROGRESS
The coordination of the study by the health departments in the five Great Lakes
States (WI, OH, MI, IN, IL) has been accomplished through constant commu-
nication and follow-through by all consortium members. As mentioned
previously, formal teleconferences are held biweekly to relay information as
well as to plan for the upcoming weeks. Immediately following the teleconfer-
ences, each State receives a detailed document with information discussed along
with important attachments (e.g., exposure history questionnaire).
Each State has acquired access to PC Wonder, a communication software
package provided by CDC. The consortium members use this package to send
information to each other. Each State also has a copy of EPI INFO, a software
package designed for data entry and analysis of epidemiological study data. The
five States have the ability to enter data into the same file, which makes merging
data easier during analysis.
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CONGENER-SPECIFIC PCS ANALYSIS FOR IMPROVED
TOXICITY ASSESSMENT
PCBs are chemical compounds having the empirical formula C12H10 nCln,
withn= 1-10. There are 209 theoretically possible PCB congeners, that can
be divided into 9 isomeric groups, and decachlorobiphenyl, each of which has
a systematic number. Because of their ubiquity and persistence, PCBs have
received considerable attention in recent years in the areas of chemical
analysis and toxicology.
The objective of this project is to develop methods to analyze PCBs and other
chlorinated environmental contaminants that will be more effective in
making health hazard assessments than currently available techniques.
Toxicity assessment differs from contamination level determination by the
requirement to quantitate levels of specific congeners, particularly those that
are the most toxic. The additional degree of selectivity required by congener-
specific analysis is often obtained at the expense of decreased sensitivity or
increased analysis cost. Recent developments in mass spectrometry and
chromatography indicate several processes that have selectivity for those
particular PCB congeners most closely related to toxicity. This project will
examine these processes and develop them as bases for new analytical
methodology for toxicity assessment. Three approaches are being explored
according to the three areas in a chromatography/mass spectrometry system
in which chemical selectivity can be exhibited:
1. Approach I is based on ion-molecule reactions carried out in the
reaction chamber of a triple-stage quadrupole mass spectrometer.
Using an oxygen-chlorine exchange reaction, the investigator has
been able to quantitate PCB congeners even when they are not
chromatographically resolved. This approach is to be improved,
implemented, and extended.
2. Approach II is based on the ionization process in the source of the
mass spectrometer. The investigator has observed a surface cata-
lyzed, chlorine-hydrogen exchange reaction that appears to only
occur with coplanar PCBs. Such conformation-dependent behavior,
particularly as it discriminates for the most toxic structures, could
provide the basis for methods specific for the toxic dongeners.
3. Approach HI involves conformation-dependent retention in con-
junction with liquid chromatographic separation and mass spectro-
metric detection. While PCB s will provide the model compounds for
these studies, polychlorinated dioxins and furans are also being
studied with the goal of including their congener-specific determi-
nation by the same general approach, if not within the same
analytical procdure.
83 Great Lakes Human Health Effects Report
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Techniques developed are being tested with samples offish harvested from the
Great Lakes and compared with the results of currently used methodology.
Improved congener-specific analytical methodology could greatly benefit the
study of congener-dependent bio-uptake and biodegradation, as well as improve
the quality and quantity of data on which public health decisions are based.
This ATSDR-supported research project is being conducted by Dr. Christie
Enke at Michigan State University.
PROGRESS AND PRELIMINARY RESULTS
Optimization of Ion Formation
The investigator's current area of research is determining the conditions under
which the formation of [M]-. ions in the source and the formation of [M-C1+O]'
ions in the collision chamber are optimized. The interest in determining these
conditions is to improve levels of detection. For the detection of extremely low
concentrations of the target compounds using MS/MS techniques, optimization
of instrumental parameters is important. The excellent detection limits of
selected reaction monitoring in MS/MS mode can be coupled with electron
capture negative chemical ionization (ECNCI) to improve the levels of detec-
tion. However, detection limits are often barely adequate for the samples of
interest, particularly for lower chlorinated compounds. ECN CI does not show
a molar response of similar magnitude across all degrees of chlorination. Present
research efforts have demonstrated an increase in molar response to ECNCI with
increased degree of chlorination (Hong et al. 1992; Erhardt-Zabik et al. 1990).
Also, the degree of electron affinity (EA) of the [M]-. ion relative to that of the
[M-H] ion and the Cl ion depends on the degree of chlorination of the aromatic
ring and to some extent the positions of this chlorination. A lower degree of
chlorination favor the formation of the Cl" ion or the [M-H]- ion that is an even
electron moiety that will not undergo chlorine exc hange with oxygen (Lopshire
and Enke 1991). This results in a decrease of the sensitivity for the oxygen-
chlorine exchange reaction.
The researcher is aware that conditions that affect the relative competition
between resonance electron capture and dissociative capture are influenced by
the reagent gas pressure, the mass of the ion of interest, and the ionization
potential of the reagent gas. The conditions that favor resonance capture over
dissociative capture as a function of reagent gas type and pressure have to be
sought in order to increase the relative abundances of the [M]~ ions and to
suppress the formation of undesirable fragment ions.
The researcher is investigating these parameters further to obtain a more
complete understanding of the optimum conditions for PCS anion formation.
Great Lakes Human Health Effects Report 84
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This will lead to conditions where assays for the 12 toxic PCBs will be
optimized.
Analysis of Samples
The greatest progress within the past year has been in the achievement of
quantitation of the 12 toxic congeners through the use of stable isotope analogs
of 3 of the congeners as internal standards. The assay has been evaluated with
respect to its selectivity, sensitivity, linearity, and reproducibility.
The linearity and the reproducibility were determined by monitoring the
reactions of the analyte and the 13C-labeled internal standard molecular anions
with oxygen simultaneously. Multiple internal standards are being used due to
the wide range of responses that the PCB congeners exhibited. Thus, 13C-labeled
congeners (13C-3,3',4,4', 13C-3,3',4,4'5, and I3C-3,3',4,4',5,5') were chosen as
internal standards. The reference standards varied from 5.0 pg/ml to 100 pg/ml
for the MS/MS calibration, and the concentrations of the 13C-labeled internal
standards were 9.6 pg/ml.
The calibration curves for the congeners were quite linear; the average correla-
tion coefficient of the linear regression between peak area and concentration was
0.990. The relative response factors for the 12 toxic congeners, with respect to
the 13C-labeled congeners, have been determined. Response factors were
calculated as(congener area/congener amount)/(reference area/reference amount).
The linear dynamic range was limited to two orders of magnitude due to
saturation of the electron multiplier.
A method has been developed and used to determine the 12 toxic congeners. The
12 toxic congeners labeled in the mass chromatograms were identified by their
retention times, the elution profile of the mass chromatogram, and the mass
spectrum. Quantitation was done using mean response factors.
1993 Activities
Gas Chromatography/Mass Spectroscopy (GC/MS) Methodology
The GC/MS methods developed during the preceeding year are being applied
to real environmental samples. The fish samples used for application of the
analytical methods developed were obtained from Professors Matt and Mary
Zabik who harvested them from the Great Lakes. The demonstated quantitation
of the method with detection limits in the low ppb range demonstrates the
feasibility of this anticipated application to real samples. Other "real" samples
to be analyzed are the samples being distributed by ATSDR for the "round
85
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Section VII - Report to Congress
robin" comparison of results from the several analytical labs used in this
program.
Work will be conducted to investigate the effect of ion source parameters in
order to improve the production of [M]~ relative to [Cl]~ for low chlorine number
congeners. Dissociative electron capture to form [M-C1] + [Cl]~ rather than
resonance electron capture to form [M]" is prevalent with lower chlorinated PCB
congeners and results in decreased sensitivity. Determining the source param-
eters that maximize [M]" production will optimize sensitivity, especially for the
lower chlorinated PCB congeners.
Experiments will be conducted to optimize the [M-C1+O] formation in the
collision cell. The effects of collision gas pressure and collision energy on the
oxygen-chlorine exchange reaction will be investigated. The search for other
molecular reactants will be commenced in an effort to find a still more efficient
reaction system.
Liquid Chromatography/tandem Mass Spectroscopy (LC/MS) Methodology
LC/MS offers possible advantages of decreased analysis time and improved
molecular ion production. Investigation into the ionization of PCBs by
electrospray ionization and separation of PCBs by liquid chromatography is
underway. The development of liquid chromatographic techniques for separa-
tion of coplanar and nonplanar PCBs and for separation of coplanar PCBs from
each other will be initiated. Work will be conducted on optimizing the
conditions for electrospray ionization of PCBs and characterizing its composi-
tional and conformational dependence.
Recently, a rapid and simple liquid chromatographic method for the isolation of
non- and mono-ortho substituted PCBs from human milk that uses porous
graphite carbon (PGC) has been described (Hong et al. 1992). The ability of
porous graphitic carbon to selectively adsorb on the basis of molecular planarity
provides a highly efficient and straight forward method to directly isolate non-
and mono-ortho substituted PCBs from real samples. These experiments
suggest that an elution chromatographic method based on the PGC compounds
could then be detected by MS or MS/MS, depending on the required additional
degree of selectivity.
Because of the limited information available with respect to the application of
these new techniques to PCB analysis, the research proposed for the exploration
of electrospray ionization and liquid chromatographic separation based on
structural conformation is more speculative than the proposed GC/MS method-
ology. Nevertheless, the researcher believes that the potential benefits of these
technologies make at least an initial exploration worthwhile.
Great Lakes Human Health Effects Report 86
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Work will be conducted to determine the feasibility of liquid chromatographic
fractionation of PCS samples into coplanar and nonplanar and/or non-ortho and
ortho fractions. These fractions could then be analyzed by LC/MS/MS or by the
previously developed GC/MS/MS methodology. It is anticipated that the
shorter run times could make LC/MS analysis valuable as a rapid screening
technique for PCBs in real samples.
MICHIGAN GREAT LAKES HEALTH STUDIES
This research project is being conducted by the Michigan Department of Public
Health, Health Risk Assessment Division Laboratory, to design, establish, and
operate a professionally creditable interlaboratory, quality assurance program
for the ATSDR Great Lakes Human Health Effects Research Program. A
number of the ATSDR-supported research projects require analytical testing of
human specimens for Great Lakes chemical contaminants, such as heavy metals,
organochlorine pesticides, and the family of synthetic poly cyclic polyhalogenated
organic compounds. Some of these compounds have no standardized test
methods. This quality assurance program will demonstrate and provide
ongoing documentation of the comparability of the analytical data generated by
the various studies.
Major objectives of the project include the following:
1. Conduct a survey of the laboratories involved in the ATSDR
Great Lakes Human Health Effects Research Program with
regards to sample size, batch size, analytes, frequency of
analysis, and estimated quantity of blank and spiked reference
sera needed for initial characterization and ongoing use as
analytical control samples.
2. Recruit additional interested agency laboratories, such as the
Centers for Disease Control and Prevention, to serve as
reference laboratories.
3. Obtain sufficient ovine sera to provide for blank and spiked
reference sera for all participating laboratories and for
extensive characterization and longevity studies to be conducted by
this laboratory.
4. Establish a database for the characterization of data and for the
ongoing control studies.
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Section VII - Report to Congress
These objectives provide for the establishment and maintenance of an
interlaboratory quality assessment program with the aim of developing com-
parative data sets on a region-wide basis. This ATSDR-supported project is
conducted by Dr. Harold Humphrey, Mr. Robert Welch and Mr. Michael
O'Keefe of the Michigan Department of Public Health, Health Risk Assessment
Division.
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