Methods

Summary

Phase 1

Collection and Housing of Kelp Species

Our team will take samples of kelp from various locations along the coast of Oregon. These samples will be tested for environmental pollutants and heavy metal concentrations. Our intention is to survey and understand the differences in populations and observe potential factors that enhance toxicity in specific areas. Each sample will be an entire specimen of kelp - with three samples taken at each location. We will travel out to the kelp forests and dive down to collect each specimen. After collection, the specimens will be housed in an ocean simulation tank at the Oregon Institute of Marine Biology (OIMB.) Once the kelp have been transferred to OIMB, they will be assayed for a variety of ecotoxins.

There is a wide spectrum of toxins that may be found in kelp. Kelp is a brown algae that readily takes up minerals, including many heavy metals such as arsenic, lead, and mercury. We may also expect to find some amount of pesticide, manufacturing, and waste pollutants, which are harmful to the kelp and environment. Other chemicals such as zinc sulfate and sodium pentachlorophenate, are thought to inhibit the reproduction process of kelp plants, thus contributing to further kelp deforestation.

Protocols for Tests

Samples of kelp will be tested for the ecotoxic compounds at the beginning of the test sessions. Over the span of one month, identical chemical tests will be taken twice a day in the morning and afternoon. Data will be documented in an excel spreadsheet in order to keep track of the changing chemical concentration in the kelp. 

Phase 2

Once we have set up a healthy control tank for living kelp plants, we can begin to record and study how long it takes it may take the kelp to detoxify from the detected pollutants.

Plant species have different mechanisms and rates of uptake and discharge for heavy metals and other pollutants. These mechanisms may include mycorrhizal association, binding plant cell wall and root excretions, metal efflux from the plasma membrane, metal chelation by phytochelatins and metallothioneins, and containment within the cell vacuole.

Protocols for Tests

We will track the metabolism of the main toxins found in Oregon kelp populations under normal conditions, without re-exposure to pollutants. Over the span of one month, chemical tests will be taken twice a day in the morning and afternoon. The primary metals we wiil sample for are Iodine, Lead, Copper and Cadmiun. 

Data will be documented in an excel spreadsheet in order to keep track of the changing chemical concentration in the kelp. At the end of this study, we should have an understanding of the time it takes for the chemicals to be metabolized from the kelp samples, and can conduct additional research into the responsible mechanisms and adaptations.

In addition to this, we will set up control tanks where we introduce exogenous metals, and examine the toxic effects on the kelp physiology. Things we will assay are toxicity threshold and microdosing (chronic exposure stress) as well as assessing time for detoxification. 

Challenges

One challenge will be keeping the Kelp alive and active monitoring of metal detoxification. Another will be the procurement of Kelp. As all of us are somewhat novice marine biologists, we may have a harder time harvesting the kelp compared to a profesisonal. One particular challenge would be, if we decided to, run DNA analysis of metal. Speficially the epigentics of metal exposure. 

Pre Analysis Plan

We plan to measure and manipulate heavy metal exposure and detoxification. We will be able to compare between geogrpahical differences (based on sampe population), different metal exposure effects and stages of development. We will be able to run ANOVA and Student T tests to analyze potential significant differences.