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Marine
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Environmental Technology
1998-2000 Projects
Molecular Biomarkers
of Chemical Sensitivity
Mark E. Hahn and Brenda A. Jensen, Woods Hole Oceanographic
Institution
A group of chemical contaminants known as PHAHs, or planar halogenated
aromatic hydrocarbons, are persistent in the marine environment.
Some of these contaminants accumulate in the blubber and other tissues
of marine mammals. As a result, certain cetaceans and other marine
mammals contain some of the highest levels of PHAHs reported in
any wildlife groups. These contaminants may contribute to marine
mammal mortality and morbidity. However, the magnitude of the risk
that PHAHs pose to the health of marine mammals is controversial,
in part because there is little direct information on the sensitivity
of these animals to PHAHs. Because legal and ethical concerns preclude
addressing these questions through direct testing of toxic chemicals
on protected animals, alternate research approaches are required.
In a previous Sea Grant study, these researchers studied PHAH sensitivity
of beluga by cloning the gene for the beluga aryl hydrocarbon receptor
(AhR) protein, which plays an important role in the mechanism of
PHAH toxicity. This project will extend that work by examining the
function of the AhR in intact cells, thus determining whether the
results obtained with the associated AhR are reflected in real cellular
events, including changes associated with toxicity. This project
will lead to a better understanding of the risk to marine mammals
exposed to PHAHs and may serve as a model for a new approach for
assessing the risk of environmental contaminants to protected species.
(R/B-151)
Biochemical Toxicology
in Cetaceans
John J. Stegeman and Michael J. Moore, Woods Hole Oceanographic
Institution
This project, related to the project described above, will attempt
to address recent scientific evidence that establishes links between
chemicals released into the marine environment and damage to fish,
birds, and mammals, including humans, by disrupting hormone action
and interfering with reproductive and developmental processes --
so-called "endocrine disruptors." These researchers have,
over the years, conducted Sea Grant studies of biochemical toxicology
in marine mammals using archived cetacean tissue samples from stranded
animals and tissue biopsies. This project seeks to expand these
efforts and to lay the foundation for a multi-institutional program
focusing on molecular, histopathological, and gross-morphological
features that may be linked to chemical effects in cetaceans. Such
a concerted effort is essential for evaluating the susceptibility
of cetaceans to different types of chemicals. Specifically, this
project will obtain and archive cetacean tissue samples for analysis
of enzyme and receptor systems relevant in chemical effects; establish
a multi-investigator program to examine the properties of molecular,
biochemical, and cellular systems that may participate in toxic
mechanisms in cetaceans; describe histological features that may
be linked to or that could corroborate biochemical results indicating
chemical effects in these animals; and determine the concentrations
of environmental chemical residues, including PAH and PHAH that
may occur in liver and other organs of ceta-ceans. (R/B-152)
Identifying Wastewater-Derived
Nitrogen in Aquatic Ecosystems: Tests of a Stable Isotope Tracer
Approach
Ivan Valiela and James W. McClelland, Boston University Marine
Program
Increasing nitrogen loading from watersheds is leading to eutrophication
of coastal waters worldwide and is considered one of the most pervasive
human-induced alteration of coastal ecosystems. To effectively manage
and monitor these ecosystems, techniques for assessing nutrient-driven
changes in coastal waterbodies must be developed. In previous Sea
Grant studies, these investigators developed a stable isotopic approach
for identifying wastewater-derived nitrogen in aquatic systems --
an approach that has proven useful for tracking septic tank-derived
nitrogen from the Waquoit Bay watershed of Cape Cod, Massachusetts,
into its surrounding estuaries. In this project, investigators will
put their approach to the test. They will determine if the approach
can be applied to a broader range of estuaries in the Cape Cod region,
in estuaries where nitrogen loads enter from sources other than
septic tanks, and in freshwater and brackish water systems. This
information -- and this innovative approach -- will be useful to
environmental managers as it will provide them with a sensitive,
economic tool for identifying and monitoring the impact of anthropogenic
nitrogen inputs on aquatic food webs. (R/M-40)
Detection and Quantification
of Live Acanthamoebae in Natural Marine Ecosystems Using Molecular
Genetic Methods
Rebecca J. Gast and David A. Caron, Woods Hole Oceanographic
Institution
This project, set to begin in 1999, will look at the free-living
amoebae Acanthamoeba. Acanthamoebae can opportunistically infect
humans. They do not generally pose a risk to healthy individuals,
other than a small number of annual cases of Acanthamoeba keratitis,
a painful corneal infection typically caused by improper care of
soft contact lenses. Acanthamoebae can be isolated from soil, salt
and freshwater (including tap water), and some marine sediments.
In the marine environment, most Acanthamoebae have been obtained
near sewage and waste dumps. Their presence at sewage dumps suggests
that they may be useful as indicators of sewage contamination. However,
analyzing the natural distribution and abundance of amoebae in general
has been problematic. This project will attempt to implement detection
and quantification methods that will provide valuable information
on the presence and distribution of Acanthamoebae and provide a
new set of tools for analysis of natural species. The development
of such methods will enable ecologists to examine the mechanisms
that affect the growth and aging of ecologically important organisms.
(R/B-147)
The following four
projects are part of a 7-project, multi-institutional initiative
supported by the National Sea Grant College Program titled, "Chemical/Biological
Interactions: Receptor Mediated Effects on Reproduction and Development
in Aquatic Species."
Fish Cytochrome P450
Genes Involved in Chemical Effects
John J. Stegeman, Woods Hole Oceanographic Institution
In this study, researchers will establish a molecular basis for
explaining and monitoring effects of anthropogenic or natural chemicals
on endocrine and developmental processes in fish. Through studying
the salt marsh minnow Fundulus heteroclitus, investigators will
clone and sequence genes for cytochrome P450 that may metabolize
and activate or inactivate chemical compounds. These studies will
provide a molecular foundation for understanding the mechanisms
and monitoring the effects of diverse chemical pollutants. The results
will have implications for and applications in ecology, toxicology
and pharmacology of fish, and could provide new approaches for screening
effluents and new chemicals for biological reactivity. (R/P-60)
Molecular Biological
Approaches for Non-Destructive Assessment of Chemical Effects on
Marine Mammals
John J. Stegeman and Michael J. Moore, Woods Hole Oceanographic
Institution
Pollutants in the marine environment, such as polynuclear aromatic
hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), dioxins,
and furans, are believed to have adverse affects on marine populations,
including marine mammals. Exposure to such pollutants may effect
the physiology of marine mammals, including reproduction, immune
defense, endocrine system functions, and, possibly, neural systems
that control social and migratory behavior. Using molecular biological
techniques, researchers in this study will focus on the pilot whale,
studying biopsy samples of free-ranging populations as well as archived
samples taken from stranded whales to evaluate the exposure of marine
mammals to harmful chemical compounds. By quantifying expression
of the cytochrome CYP1A gene, a sensitive biomarker of hydrocarbon
effects, in the livers and other organs of the pilot whale, investigators
will be able to characterize metabolic activity. The methods established
in this study will provide a non-destructive means to study compounds
that may pose a threat to fragile marine mammal populations. (R/P-61)
Impact of Environmental
Contaminants on an Aquatic Bird Population
Mark E. Hahn, Michael J. Moore, and Constance A. Hart, Woods
Hole Oceanographic Institution, and Ian Nisbet, I.C.T. Nisbet and
Co.
Many questions surround the extent to which dioxin-like compounds
and chlorinated pesticides disrupt endocrine functions -- such as
reproductive and developmental processes -- in humans and aquatic
animals, including birds. This project seeks to examine abnormalities,
including gonadal feminization, in Common Tern embryos from two
coastal Massachusetts sites with different levels of environmental
contamination. Using chemical, biochemical, and immunohistochemical
methods as well as cell-culture bioassays, researchers will measure
exposure of the birds to a variety of environmental pollutants.
Through a combination of field and laboratory-based studies, investigators
hope to assess possible relationships between health effects and
specific contaminants, including dioxin-like compounds and environmental
estrogens. (R/P-58)
Identification of Bioactive
Marine Natural Products Using a Fish Culture Bioassay
Mark E. Hahn, Woods Hole Oceanographic Institution
Marine organisms produce a variety of organic compounds (marine
natural products), some of which display useful biological activities.
Many of these products are structurally related to dioxins, PCBs,
and other anthropogenic compounds that act through specific receptor
mediated mechanisms. Investigators in this study will use a newly-devised
cell culture bioassay system to determine marine natural products
that stimulate or inhibit the same biochemical systems that respond
to dioxins. Exploring the ability of marine natural products to
mimick the effects of dioxins and PCBs will be important to understanding
the biochemical and physiological changes that have been observed
in animals living in contaminated environments. In addition, some
of these natural compounds may be useful as future research tools
or as pharmaceutical agents. (R/B-124)
The following project
is part of a National Strategic Initiatives (NSI) competition in
Marine Biotechnology, made possible by a special National Sea Grant
College Program award.
Detection of Harmful
Algal Species Using Molecular Probes: Field Trials
Donald M. Anderson, Woods Hole Oceanographic Institution
Over the past two decades the economic and public health impacts
from harmful algal blooms (HABs) have increased dramatically throughout
the world. One result of this expansion is that regulatory officials
and the fishing industry are faced with a broad array of affected
species, spanning all levels of the food chain, many of which can
be contaminated by several different toxins. These changes have
forced a major reevaluation of strategies to monitor seafood products
for marine biotoxins. Present techniques to identify phytoplankton
species and to count toxin rely on manual microscope techniques
that are time consuming and require trained specialists. This project
represents the last phase of ongoing efforts to develop molecular
probes and assay systems for several key HAB species. By refining
and field testing nucleic acid-based probe assays that can be used
in the laboratory and the field by personnel with limited technical
expertise, this project will contribute an accurate, fast, and reliable
tool for private, state, and federal monitoring programs for HAB
species. (R/B-146)
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