Lab 5: River Experiment Part I
Developing a Fundable Proposal Concerning American River Ecology
Introduction and Brainstorming Session
What you should know after this lab:
• How experiments under laboratory conditions can be used to support more
broadly-based field studies
• The biodiversity of American River aquatic macroinvertebrates
• The impact of human-associated environmental nitrogenous inputs
• How eutrophication can impact an aquatic ecosystem
• How differences in aquatic macroinvertebrate populations can be used to
indicate pollution levels
Skills you will reinforce from previous exercises:
• Hypothesis development using supporting literature
• Draw on and incorporate ecological information gained during Laboratory- and
Activity exercises to support hypothesis- and project development
• Organizing electronic data sets appropriately to provide supporting analysis for
• Collecting, analyzing, and interpreting data using MS Excel
The American River Parkway
The American River Parkway is a diverse ecosystem, which is home to over 1,200
species of plants, birds, reptiles, mammals, fishes, invertebrates, and other organisms.
Because this river and its accompanying ecosystems provide many resources to nearby
urban areas such as Sacramento, interactions between humans and these areas can
have a substantial environmental impact. There are many human-driven factors such as
fire, mowing, and road maintenance that can affect the species composition of any
particular area including those surrounding the American River. For example, soon you
will assess the impact of human-derived disturbances using measures of local plant
communities along the American River Parkway near CSUS to validate the Intermediate
Disturbance Hypothesis. If the data you collect validates the hypothesis, it will suggest
that human-derived environmental disturbances impact plant species richness. It can
be extrapolated then, that similar effects likely would be observed for animals as well.
Similar research often has been used to assess other human-derived
disturbances such as chemical runoff into watersheds. In your Activity sessions, you will
be introduced to a research study concerning the effects of three farming types
(organic, integrated, and conventional) on aquatic macroinvertebrate populations in
streams in New Zealand (Magbanua et al. 2010). In that research study, the authors
determined that farming type affected aquatic macroinvertebrate species composition.
In addition, the authors stated that there may have been many factors within each
farming type contributing to differences in species richness such as physiochemical and
the nutritional compositions of each sampling site.
Many times, a large research project such as that by Magbanua et al. (2010)
leads to many more questions than answers. Therefore, researchers often begin to
eliminate unimportant factors by isolating and evaluating selected variables. Over the
last several years, students in Bio 1 have conducted a laboratory experiment, which
evaluated the potential effects of fertilizer runoff (added nitrogen) on aquatic
macroinvertebrates found in the American River. Fertilizer treatments emulated the
total nitrogen concentrations determined for conventional farming areas previously
described (Magbanua et al. 2010). The following methods for the experiment in Bio 1
are given below for reference:
Materials and Methods
To assess potential impacts of fertilizer runoff on American River aquatic
macroinvertebrate populations, a river ecosystem was simulated using 10-gallon (~38-L)
glass aquariums under laboratory conditions. All aquariums were fitted with plant and
aquarium bulbs with light/dark (12L:12D) durations maintained by timers. Plastic tubing
affixed to the aquariums provided oxygen from permanent fixtures on the laboratory
benches. The activity of these air tubes also approximated water movement in the area
where our substrates were collected in the American River. Approximately 5 gallons
(~19 L) of water and 5 cm of sediment were collected and added to each aquarium.
Each Lab was assigned one Control Aquarium and one Treatment Aquarium. On
each sampling day, kick-netting, a technique often employed by researchers wishing to
sample aquatic invertebrates, was emulated in each aquarium. Briefly, the top 1-cm
portion of the sediment in the Control Aquarium was completely disturbed with a small
tool. One member from each lab bench collected disturbed water samples with a 200-
ml glass jar just below the water surface.
The 200-ml sample was sieved using nylon stocking and the resulting debris was
distributed onto three fingerplates. A dichotomous key was provided to identify and
count any macroinvertebrates encountered in individual samples using the dissecting
microscope. This process was repeated similarly for the treatment aquarium, and the
entire sampling day was repeated approximately 3 weeks after fertilizer was added.
During the second sampling day, the potential change in algal growth and thus,
any potential effects on aquatic macroinvertebrates that might be attributed to
eutrophication was assessed indirectly by use of a spectrophotometer. Briefly, each
group collected a small sample (~10 ml) from their 200-ml glass jars and measured the
level of absorbance using a spectrophotometer for chlorophyll a (665 nm). Higher
absorbance readings indicated greater chlorophyll a levels and perhaps indicated,
greater algal growth.
After sampling was completed for all aquariums on Sampling Day 1, fertilizer was
applied to the treatment aquarium. The selected dose emulated the findings of
Magbanua et al. (2010). Briefly, a commercially-available liquid fertilizer (10% nitrogen)
was applied at the average rate previously-determined for streams near conventional
(High Treatment) farms.
Now that you have been introduced to the experiment that has been conducted
for several semesters, it will be your job to use that information to support a new, but
related study that you propose. Keep in mind, that your proposal can be about any
organism, in any American River microhabitat that you can imagine, and that our
previous work should be used to justify your proposed “next step.”
You and your bench mates will spend two Laboratory sessions beginning today,
discussing and designing a potential project that ultimately, will be presented in written
form as your last lab report. Your task will be to formulate a proposal that could be
submitted to a government agency in an effort to gain funding to support it-in essence,
you will be writing a mini-grant. Although we encourage collaboration regarding project
development, this report will be generated on an individual basis and must contain all
sections included in your previous submission regarding phenotypic plasticity
(introduction, materials and methods, potential results, discussion/conclusions).
However, it will be presented as a potential project, rather than one you have
Over the past few weeks, you have been introduced to and used a variety of
biological study tools such as developing and testing hypotheses, taking measurements
with calipers and scales, using microscopes, and conducting data collection and analysis,
to name a few. In the next few weeks and until the due date for your proposal (your
instructor will inform you about their chosen due date), you will be introduced to
several ecologically-themed exercises, including testing the intermediate disturbance
hypothesis, and studying the life histories of several plant and fungal lineages. You will
be required to consider these things you have learned and incorporate some of them
into your proposal.
Your instructors will discuss with you the purpose of the previous River
Experiment regarding the potential effects of added nitrogen on American River
macroinvertebrates. In addition, they will provide you with a dataset from this
experiment, which you will use to provide a supporting analysis for your proposal. Your
task will be to develop a brief, two-page, potentially fundable project, which builds on
the data/findings collected by previous Bio 1 students, and incorporates
skills/techniques/concepts you have learned from at least two Laboratory sessions this
semester. In addition, your project must be supported by in-text citations of both peerreviewed
articles you have discussed in Activity.
The possibilities for your proposal are seemingly infinite. Therefore, your
instructors will be assessing primarily your ability to demonstrate mastery in hypothesis
development, appropriate experimental design and analysis, and interpretation of
Below is a list of requirements that your project proposal should incorporate.
These are required items that are in addition to the items that are listed in the grading
rubric for your Laboratory Reports. For each of the items on the list below, for which an
attempt is not made, a 10% reduction in your overall score will be made (e.g. missing all
components below would result in a 70% reduction in a perfect Lab Report rubric score
of 25pts, resulting in a score of 7.5)
1.) Proposal contains pertinent sections (Intro, Materials and Methods, Potential
2.) Proposal is two pages in length.
3.) Proposal provides an analysis of previously-collected data including statistical
methods and supporting graph(s).
4.) Proposal contains an appropriate hypothesis and rationale.
5.) Proposal includes two or more skills/techniques/concepts you have learned
from at least two Laboratory sessions.
6.) Proposal includes appropriate in-text citations from both previously
discussed journal articles (from Activity sessions).
7.) Proposal includes an appropriate interpretation of potential findings (e.g.
what does this project mean for the community with regard to the American
Use the remaining portion of this page as a place to write/draw potential
avenues for your study design. As you discuss potential target organisms for your study,
remember to consider things such as potential sampling techniques including
randomization, how you will employ statistical testing with your knowledge of t-tests,
and how it will relate to something meaningful with regard to the American River.
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