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Melissa Chan
Professor Daniel Enquobahrie
Professor Victoria Holt
EPI 221
December 15, 2016

Role of Chronic, Low-Level Maternal Exposure to Domoic Acid

in Offspring Adverse Neurodevelopmental Outcomes

Introduction

In November of 1987, hospitals in the New Brunswick and Quebec Canadian provinces received

a wave of patients presenting with rapid onset confusion, disorientation, and memory loss (Costa et. al.

411). A total of 250 such cases were identified, with the most severe including seizures, coma, and four

deaths (Costa et. al. 411). All symptoms were linked to recent consumption of mussels harvested from

Prince Edward Island (Costa et. al. 411). Further investigation revealed these mussels were contaminated

with domoic acid, a potent neurotoxin, and the condition was named amnesic shellfish poisoning (Costa

et. al. 411).

This incident was the first to identify the connection between domoic acid (DomA) and amnesic

shellfish poisoning (ASP). It catalyzed active research into the mechanism of DomA neurotoxicity, as

well as the development of FDA regulations on commercial, recreational, and subsistence harvesting of

at-risk seafood based on seasonally measured toxin levels. However, nearly all of the research conducted

and regulations implemented were focused on protecting adult consumers from sudden, high-dose toxin

exposures, such as in the 1987 Eastern Canadian incident. More recent studies have begun to indicate that

a long-term, low-dose exposure could be just as dangerous, particularly for children.

Unlike most commercial and recreational seafood consumers, subsistence harvesters rely on

seafood as a staple part of their daily diet, and are consequently at much higher risk for DomA exposure.

As part of cultural tradition, populations like the Native American communities along the Washington

State coast regularly consume at-risk seafood, such as razor clams and Dungeness crabs. This means that

even under current FDA regulations, infants, children, and adults in these communities are exposed to

chronic, low-doses of DomA. Initial studies in animal models have shown that children are more
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susceptible to low-doses of the toxin, and that fetal neurodevelopment can be adversely affected by low-

dose maternal exposure (Costa et. al. 413).

In this paper, I will investigate the effects of DomA on childrens neurobiological and cognitive

development, with a specific focus on Washington State coastal Native American populations where

reproductive-age women are exposed to chronic, low doses of DomA via regular shellfish consumption.

Based on recent animal model research and current human epidemiological studies, I will also evaluate

whether the current FDA regulations on safe daily intake levels are protective enough to address these

concerns, and suggest recommendations for future research.

Background: Domoic Acid & Neurotoxicity

DomA is produced by marine plankton from the genus Pseudo nitzschia (Costa et. al. 410).

Shellfish such as razor clams, crabs, mussels, and scallops become contaminated when they feed on these

plankton, and cooking seafood before human consumption does not reduce DomA toxicity (Costa et. al.

410). Figure 1 shows a map of high-risk sites along the Washington State coastline (Lewitus et. al. 138).

Fig. 1 Washington coast line with sites of highest

concentration of DomA contaminated shellfish. Numbers with
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no units: g/100g shellfish meat. DomA: ppm. (Lewitus et.

al. 138).
DomA acts on neural networks by mimicking the endogenous excitatory neurotransmitter,

glutamate (Costa et. al. 413). High dosage was defined in the 1987 incident as greater than 2.0 mg/kg bw

(Costa et. al. 411). At this level, DomA over-excites target glutamate receptors, resulting in a series of

events ending in elevated cellular oxidative stress and neuronal necrosis (Costa et. al. 413).

Neuropathological studies on the four patients who died from ASP in 1987 found neuronal necrosis in the

hippocampus and amygdala brain regions where the target glutamate receptors are represented in high

density (Costa et. al. 411). These regions are responsible for memory, decision making, and emotional

reactions, which both explains the symptoms of ASP and indicates the potential cognitive developmental

complications in children exposed to DomA. In fact, studies with animal models show that both pre- and

post-natal exposure to DomA causes hippocampal damage in offspring, as well as seizure disorders and

persistent behavioral abnormalities (Costa et. al. 413).

Public Health & MCH Importance

The FDA bases regulation standards on a combination of regular phytoplankton surveys and

sampling of indicator species that assess toxin levels throughout the year (Lefebvre et. al. 223). Currently,

human tolerable daily intake (TDI) is set at 20 g/g of shellfish tissue, and harvesting bans are

implemented if sampling reveals toxin levels higher than this (Lefebvre et. al. 223).

However, this TDI level is based on the standard 60 kg human male model, and may be too high

for sensitive subpopulations (Angus 103). Studies with rat models found that fetal and neonatal rat pups

were two orders of magnitude more sensitive to DomA toxicity than adults (Costa et. al. 417). This is

most likely due to the greater susceptibility of the developing brain to neuronal necrosis from elevated

oxidative stress (Costa et. al. 413). Additionally, DomA was detected in fetal rat pup amniotic fluid, as

well as in maternal milk even after it was no longer present in maternal urine (Costa et. al. 412). If these

findings are generalizable to humans, they indicate that infants could continue to be exposed to DomA
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even after the toxin has cleared the mothers system, thereby contributing to a chronic, low-dose exposure

to DomA over a lifetime that begins even before birth (Angus 107).

The implications of these findings are particularly salient and concerning for Native American

communities along the Washington State coast, who have a strong cultural reliance on razor clams as a

staple protein in daily diet (Lefebvre et. al. 224). The table in Figure 2 describes data collected to derive a

chronic seafood consumption rate between different groups of consumers in Washington State (Angus

111). The Native American Subsistence Fishers group consumes a mean rate in g/day that is almost 30

times higher than the General Population group, and is therefore at a 30 times higher risk of daily

DomA exposure (Angus 111).

Fig. 2 Chronic Seafood Consumption Rate Between Population Groups (Angus 111)
Group Consumption Rate (g/day)
General Population 11.3 19 mean
Recreational Marine Fishers 5.6 24 mean
Native American Subsistence Fishers 540 mean

Furthermore, Figure 3 shows a plot of DomA concentration ( g/100g shellfish tissue) from

1957 to 2012 along the Washington State coast (Lewitus et. al. 141). The red line indicates levels that

correspond to the FDA TDI of 20 g/g of shellfish tissue. It is clear that the frequency of high toxin

level episodes has been increasing since the 70s.

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Fig. 3 Plot of DomA concentration ( g/100g shellfish tissue) from 1957

to 2012 along the Washington State coast. The red line indicates levels that
correspond to the TDI of 20 g/g shellfish tissue. (Lewitus et. al. 141).
Current Knowledge

Almost all research on DomA neurotoxicity has been conducted on animal models, and of these

only a handful investigated the effects of low-dose maternal exposure on fetal neurodevelopment and

infant cognitive outcomes. The table in Figure 4 below describes key findings on behavioral and

structural effects of in-utero and postnatal exposure to DomA in several mouse and rat experiments.

Fig. 4 Key findings on behavioral and structural effects of in-utero and postnatal exposure to
DomA in various experiments with mouse and rat models (Angus 68-77)
Animal Model DomA Exposure Route Effects Reference
Myelination failure
3 intraperitoneal in cortex
injections of mother at
Exposure Increased anxiety-
Mice early embryonic, early Tanemura et. al.
In-Utero related behaviors
fetal, and late fetal time
points Impaired learning
and memory
Single dose
subcutaneous injection
Rats Motor seizures Wang et. al.
of pup on postnatal day
Exposure 7
Postnatal
Daily intraperitoneal Mossy fiber
Rats injection of pups from (neuron) sprouting Bernard et. al.
postnatal days 8-14 in hippocampus

In general, studies conducted in-utero found that DomA crosses the placenta to affect fetal brain

structures, resulting in behavioral effects later in adulthood (Angus 69). Additionally, they showed that

DomA exerts greater effects when exposure occurs later in gestation (Angus 69).

Studies conducted postnatally found that neonatal rat pups were most sensitive to DomA during

their first two weeks of life, which represents a critical period for brain development (Angus 74).

Additionally, common structural effects included mossy fiber neuron sprouting in the hippocampus,

which has previously been associated with epilepsy and head trauma in humans (Angus 80).

Ultimately, each study involved exposing either the maternal or offspring generation to levels of

DomA low enough to avoid physical sickness. This enabled them to show that even when no immediate
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maternal or offspring toxicity was observed, significant behavioral and structural effects in offspring

could manifest later in life (Angus 69).

Currently, the National Institute of Environmental Health Science is conducting the first human

epidemiological study on long-term, low-dose DomA exposure with three pacific northwest Native

American tribes. The Communities Advancing the Studies of Tribal Nations Across the Lifespan

(CoASTAL) Cohort includes the Quileute Indian Nation, the Makah Tribe, and the Quinault Indian

Nation (Tracey et. al. 10). The study began in 2005 and follows a community-based participatory research

model (Tracey et. al. 10). Figure 5 illustrates a map of the study region, which overlaps with the map of

high-risk DomA exposure sites shown in Figure 1 on Page 2 (Tracey et. al. 10).

Fig. 5 Map of CoASTAL Cohort study region, which

overlaps with high-risk DomA exposure sites shown in Fig.
1 on Page 2 (Tracey et. al. 10).
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Wave 1 of the study followed participants for five years from 2005 to 2010 (Tracy et. al. 10). It

had a total of 678 members divided into four age groups: 32 infants (9-16 months), 108 children (6-12

years), 507 adults (18-64 years), and 31 geriatrics (65+ years); both sexes were represented about equally

within each group (Tracy et. al.10). Participants were interviewed during their first visit to obtain baseline

assessments for medical history and cognitive screening (Tracy et. al.11). Dietary assessments were

obtained with the Block Food Frequency Questionnaire, which describes frequency of consumption for

specific foods and food groups (Tracy et. al.14). They were also given a shellfish consumption dietary

intake questionnaire, confounding exposure questionnaire, and a take-home one-day food record to be

returned within five days of the visit (Tracy et. al.14).

Subsequent participation in the study involved completing two one-day and one two-day dietary

record every four months (Tracy et. al.14). Additionally, investigators sampled shellfish from eleven

beaches along the Washington State coast to measure DomA levels throughout the study period (Tracy et.

al.14). These data were combined with the participant dietary records to ascertain exposure level, which

was defined as the average number of razor clams consumed per month in the last year, multiplied by the

mean measured DomA concentration in razor clams at all beaches from which the participant ate clams

last year (Tracy et. al. 15). From this, three exposure levels were identified: non-consumers, low-

consumers (less than 15 clams/month), and high-consumers (more than 15 clams/month) (Tracy et. al.15).

It is important to note that all exposure levels, including the high-consumers group, are still under the TDI

of 20 g/g of shellfish tissue.

Preliminary analysis of cognitive impacts after Wave 1 was just published in July 2016.

Assessment with participants in the adult age group found that when using the non-consumer and low-

consumers as reference, high-consumers performed worse on memory measures and had a greater number

of depression symptoms (Grattan et. al. 6).

Recommendations for Future Research

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The greater susceptibility of young pups to DomA neurotoxicity in the rat and mouse models,

combined with the elevated mean rate of consumption of high-risk seafood in Native American

populations suggests that the current FDA TDI level of 20 g/g of shellfish tissue is not protective

enough for sensitive subpopulations. Normed on the standard 60 kg white male model, this TDI level is

not representative of children or Native American communities, let alone children in Native American

communities. Further research in both animal models and epidemiological studies is necessary to

characterize the effects of DomA exposure on fetal and infant neurodevelopment, in order to develop a

revised TDI.

All published literature on antenatal and postnatal DomA exposure in animal models have studied

exposure via direct injection in either the pregnant mother or newborn pup. Since the primary route of

DomA exposure in humans is through oral consumption of contaminated shellfish, it would be useful to

conduct animal model studies with the maternal generation exposed in this method for a period of time

before pregnancy as well as throughout pregnancy. This would enable investigators to determine if the

behavioral and structural effects seen with exposure via injection also occur with exposure via oral

consumption.

The currently ongoing CoASTAL Cohort represents a prospective, longitudinal, human

epidemiological study that encompasses age groups representative of a lifespan. However, infants and

children comprise a much smaller proportion of this broad spectrum of participants (32 infants and 108

children, in comparison to 507 adults). In order to more effectively study the effects of DomA exposure

on maternal-child health, a separate cohort focused on women of child-bearing age and young children

could be composed and studied in parallel. Additionally, after further research with animal models is able

to more definitively link DomA exposure to specific neurodevelopmental and cognitive outcomes, it

could be relevant to conduct a retrospective case control study. Looking at older children in the

CoASTAL Cohort communities with the specified cognitive outcomes would enable a comparison of their

exposure levels and that of their mothers while pregnant with them to outcome severity.
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Finally, the CoASTAL Cohort study was formed in part because of concerns with the increasing

frequency of high toxin level episodes and harvesting closures over the past few decades, as discussed

with Figure 3 on Page 4. Additional research could investigate potential causes of these increased algal

blooms and determine if there is a factor of human agency that could be addressed as a method of

primordial prevention. Furthermore, the development of a reliable biomarker for DomA exposure would

assist secondary prevention efforts to mitigate the development of cognitive impairments.

Conclusion

Research on the effects of long-term, low-dose DomA exposure on fetal neurodevelopment and

infant cognitive outcomes is still relatively new. Experiments with animal models and the current

CoASTAL Cohort human epidemiological study have begun to characterize correlations between

exposure and mild cognitive impairments; however, further investigation is needed to support these

findings specifically in relation to maternal-child health. Ultimately, the studies described and

recommendations proposed represent the first step to addressing a public health concern that

disproportionally impacts already marginalized subpopulations in our local Native American

communities.
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References

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