Microcosm Design for Procambarus clarkii

BSEN 5510
December 14, 2018
Group # 2 - Crawfish Crew
Harris Pride
Tel Schieler
Samuel Tatum
Abstract
In order to understand the interactions, present within an ecosystem, a closed system was
designed to support and sustain Procambarus clarkii and its related organisms. The objective of
this experiment was to produce a sealed tank that could recycle nutrients for a given crawfish
mass for as long as possible. Attention in tank design for crawfish health was focused on
physical structure (abiotic conditions) and water quality (biotic factors). The abiotic conditions
included tank divisions and hiding places, water movement, and local pond sediment. Biotic
factors were introduced in local and non-local plant inclusion and from microbial life from pond
water, plants, soil, and those present with P. clarkii. The above conditions allowed for a crawfish
mass of 50.7g to be supported for a 7-day closure period.

Project Goals and Objectives

The objective of this experiment was to produce a sealed tank that could recycle nutrients
for a given P. clarkii mass during a minimum trial period of 7 days. Once the tank was closed, no
materials were allowed enter the tank for the remainder of the trial period. Procambarus clarkii
ecology was considered in order to get a better understanding of the components necessary for
the crawfish to thrive. To achieve this, the design goal was to maximize crayfish health by
creating conditions that simulated their natural habitat. This was done by focusing on the
physical structure (abiotic conditions) and water quality (biotic factors) within a 10 gallon tank.
The objective for abiotic conditions was to provide tank divisions and hiding places to
simulate crawfish tunnel and minimize stress. Water movement and local pond sediment were
also incorporated to mimic a stream habitat. Biotic factors were introduced in local and non-local
plant inclusion, and from microbial life from pond water, plants, soil, and those present on P.
clarkii. Microbial and plant inclusions were designed to cycle nutrients within the system so that
levels would remain normal. Careful consideration was given to dissolved oxygen and
ammonium concentrations since these were supposed to be the forms of toxicity.
A budget of $50 was allowed for additional required items. Not included in the budget
were most of the abiotic factors such as the fish tank, lights, lid, and some the tank sediment and
rocks. The objective was to completely the project under budget by designing a tank and
monitoring system as efficiently as possible.

Technical Approach
A two-tank design was chosen in order to treat and recycle nutrients within the system.
One tank would serve as the crawfish dwelling tank and the other as a bioreactor tank. The
bioreactor was designed to ensure good quality of water by using constant light and different
plant species (local and non-local), algae and microbes from pond water to remove nitrogen and
resupply oxygen. The bioreactor contains native aquatic plants (all of which are unidentified),
soil, and pond water in order to drive some of the biological processes necessary for crawfish
health. High plant biomass may be added to the bioreactor tank to increase oxygen production,
but not increase ammonium production from excess waste generation. Both tanks are under
separate constant light sources, but the bioreactor tank light source is stronger. Water is drained
from the surface of bioreactor tank back into the dwelling tank.
The two-tank system was considered to be best option for supporting a healthy crawfish
population by maintaining good water quality. Before the one-week closures there were 1-day
and 2-day trials in order to ensure that are ecosystem is capable of sustaining the crawfish. These
shorter trials allowed for small modifications to be made to the system and for comparison of
tank conditions over time.

Design Aspects and Details

The crawfish tank was designed to be two levels in order to keep the crawfish away from
each other due to their aggressive nature. The dwelling tank was also equipped with PVC to
allow the crayfish to hide and feel safe. The level division was introduced by adding two
horizontal acrylic sheets. By heating up one of the acrylic sheets about two inches from one end,
we were able to bend the sheet to form a ramp. The second acrylic sheet was then hot glued to
the bend to extend the ramp to the bottom of the tank to allow the crayfish to crawl up it. We
then drilled small holes along both acrylic sheets to allow the flow of water from one level to
another. We also took a razor blade found in the lab and scored the acrylic sheets in hope that it
would allow the crayfish to have more traction on the ramp and to create more surface area for
algae growth. Supports for the second level were made with ½” PVC pipe and placed on all four
corners, one of the supports fell off while placing the contraption in the tank, but it seemed
sturdy without it, so we decided it did not need to be reinstalled. The second level also allowed
the crawfish to elevate themselves above the water level, which was often observed. Figure 1
shows a good view of the second level. Around 40g of hornwort was placed in the living tank for
the crawfish to consume and for additional oxygen production.

Figure 1. Dwelling tank after 7-days of closure - Good view of the second story and overall
dwelling tank layout.
To incorporate a two-tank design, we needed to connect the tanks in a way that would
have the water constantly exchanging between the two. To do this we used a small adjustable
pump that was set to 40 GPH and used it to pump water from the dwelling tank to the bioreactor
in order to move the oxygen poor, ammonium rich water into the bioreactor. The bioreactor was
filled with approximately 450g of various aquatic and semi-aquatic plants that would then be
fertilized by the ammonium rich water and re-oxygenate it before sending it back to the dwelling
tank. We used a ½” diameter vinyl tubing for the water to be transported in. The tubing running
from the pump went into the opening at the top of the bucket and was zip tied to the bucket
handle to keep it in place.
In the bioreactor, we had an overflow system on the opposite side from the pump outflow
tube that would take water from the top of the bucket and send it back down to the dwelling tank
utilizing gravity, since the bioreactor was placed on a shelf about one foot above the top of the
dwelling tank. A hole was drilled through a rubber stopper, and the overflow tube was pulled
through it. The stopper was then jammed inside the hole drilled in the side of the bucket for the
tube to pass through to keep the bucket from leaking. Finally screen filter systems were installed
on both the overflow tube and around the pump intake to keep the system from becoming
clogged. Figure 2 shows the layout and design of our microcosm, and Figure 3 shows a picture
of our finished build. Table 1 shows a list of the materials used.

Figure 2. Schematic of the two-tank design with its major design components
Figure 3. The crayfish tank (bottom) and the bioreactor (top) exchanging water between them
and working as designed.

Table 1. A list of the materials used along with their cost and source.
System Performance
For the 1, 2, and 7-day closure trials, measurements were taken from the tank
immediately before and after each trial. Measurements included pH, dissolved oxygen,
ammonium, and temperature readings. For the 7-day trial, measurements were also taken from
the tank since the reactor tank had seemingly reached steady conditions. These records are
shown below in Tables 2 and 3 and depicted graphically in Figure 3. Comparison between the
trials (especially earlier) may not be valid since the system was not allowed sufficient time to
settle.
Dissolved oxygen was the constraint that was considered most closely. It should be noted
that it improves as the trials become longer. This implies that the photosynthetic rate of the
plants increases since no plant biomass was added. For the 2 and 7-day trials, DO was even
above ideal amounts. Most notable of the measured conditions was the ammonium
concentrations, which were 400% higher than the ideal conditions for the crayfish (Figure 3).
During the 7-day test, the crayfish appeared to be healthy. Further consideration should be given
to the amount of time crayfish may be exposed to such a concentration with no adverse effects.
Temperature and pH both remained fairly constant and were near ideal conditions.

Table 2. Recorded bioreactor and dwelling tank comparison for 7-day closure with ideal values
Tanks Trial pH DO (ppm) NH4+ Temperature
(ppm) (C)

Bio Tank Pre-7 day ~7 6 NA 23

Post-7 day 7.2 5.75 4 24.4

Pre-7 day 7.2 7.66 3.5 23

Dwelling Tank
Post-7 day 7.3 5.95 5 24.8

Ideal 7 5 1 25
Table 2. Actual and Ideal conditions from 1 and 2 day closures
Trial pH DO (ppm) NH4+ (ppm) Temperature (C)

Pre-1 day ~7 6.6 ~3 22

Post- 1 day 7.2 3.4 ~3 23

Pre-2 day 7.2 7.66 3.5 23

Post-2 day 7.3 5.5 5 24.8

Ideal 7 5 1 25

Figure 3. Percent deviation from ideal conditions for 1, 2, and 7 day trials with consideration to
dissolved oxygen, ammonium, and temperate
Figure 4. During closure (7-day) P. clarkii uses divider to go above water level

Figure 5. During closure (7-day) P. clarkii hides in PVC behind plant debris

Examining the remaining crayfish closely reveals that little white worms have embedded
themselves in the crayfish. These are annelids and actually form a mutualistic companionship
with the crayfish (Wells, 2014). It is assumed these are Branchiobdella astaci, however, further
evaluation should be performed for verification. The worms eat away dead tissue, parasites, and
epibionts, which results in the cleaning of the crayfish’s gills. The worms in turn get a good food
source. Crayfish even grow larger, quicker when the worms are present. The worms were most
likely brought in from the sediment and plants used in the bioreactor since we did not notice
them when we first picked out our crayfish. These worms can be seen in Figure 6.

Figure 6. During closure (7-day) P. clarkii feeds near the pump intake filter and B. astaci can be
seen.

In Figure 7 it was seen that after the selected local plants (~450g), local pond soil (1kg),
and associated organisms were allowed sufficient time to grow and settle into the bioreactor
tank, additional macroinvertebrates were scene to be supported. At least 5 snails were counted
after the end of the week-long closure. These may be identified in order to further evaluate the
water quality and system ecology. If these snails are considered a keystone species, it will further
support the effectiveness of the microcosm design. Additionally, new growth was seen on many
of the plants present. Some of the selected plants had been killed in from shock during the
transfer; however, after around a month of normalcy some of the plants began to thrive and
showed signs of high growth. This can be seen in Figure 8.
Figure 7. After 7-day closure, plants are alive and growing in bioreactor and a snail is present

Figure 8. Inflow (left) from the dwelling tank and filtered outflow (right) on opposite sides of
bioreactor tank
Challenges and Pitfalls
Biological Challenges
One of the major unknowns that remained unresolved was the photosynthetic rates of the
selected plants, particularly those that were locally collected. Even if the plant species were
known, further analysis would be required in order to obtain specific nutrient uptake rates. The
initial 1 and 2-day trials were used comparatively in order to approximate if the system would
supply sufficient oxygen through photosynthesis. Since dissolved oxygen was not a significant
obstacle in any of the trials, further analysis of individual plant photosynthetic rates was not
necessary; however, if system sizing based on our model is used in the future, plant nutrient
uptake and photosynthetic rates would be necessary for tank sizing and carrying capacity
estimations.
Additionally, system performance may have been increased by allowing more time for
the included organisms to settle from transport and incorporation shock. The tank was allowed 2
weeks to acclimate. This included tank rocks and sediment, PVC and shelters, pump, and
crawfish pond water and related organisms. The bioreactor tank was allowed only a few days to
acclimate before testing. Since the relations present in such a system are highly variable, the
reactor tank was likely still unsettled by the first trials. For instance, at the start of the one-day
trial, the water was still very turbid. By the end of the 2-day, sediment was settled. Factors such
as this probably affected and skewed results. In further analysis the system should be given
sufficient time to reach pseudo-steady state conditions. Otherwise, the trials will be less useful
for comparison.

Design Challenges
The greatest headache with our microcosm design was figuring out how to seal the drilled
hole in the bioreactor bucket. The hole was needed to use gravity to move the water instead of
purchasing another pump. We attempted to use a mix of hot glue and PVC glue initially, but the
PVC glue did not stick to the plastic that the bucket was made out of and it required to much hot
glue to seal. The second idea was to use waterproofing caulk. This did seal the leak, but it did not
provide any rigidity to the piping. While installing the overflow filter, the seal became weak
from pipe movement and failed. The final idea was to use a rubber stopper like a cork in a barrel.
We had to drill a larger hole in the bucket to fit the large stopper, and then we drilled a hole in
the stopper itself to allow for the vinyl tube to be pushed through after coating it in Vaseline. We
did not use any glue or sealant on the stopper, it is being held in place by the force of the hole’s
edges acting on the stopper and the water pressure pushing the stopper against the side of the
bucket.
Another design challenge we faced was matching the flow of the pump with the free-flow
of the water leaving the bucket. Our group’s original design was to match the diameter of the
drilled hole with that which would equal the 40 GPH pump when free flowing from gravity. This
would have been difficult, and due to the limited drill bit options available to us, we made the
decision to change the design to have a overflow system with a larger than necessary hole being
drilled. The overflow system works by having only the pump flow determining the flow of in
and out of the bucket to keep a constant volume. This system utilizes the conservation of mass. It
can be seen in Figure 9.

Figure 10. The overflow system working (left is the overflow tube). Water being pumped from
the crayfish tank is from the tube on the right. The screen around the overflow tube is explained
in the next paragraph.

This design worked well but became clogged after a short while. To keep from debris
entering the tube, a debris screen was added around the top of the overflow tube. Some clogging
was experienced on the screen; however, water level was not greatly affected. The screen may
eventually clog as debris piles around it, but the screen sticks far out of the water, so the water
level is able to rise a considerable amount while still having the system functioning properly. As
the debris clog the filter, the water level will rise and then be able to flow back into the dwelling
tank. We do not think this will be an issue as the filter allows the water level to be somewhat
flexible and the decomposition of the debris blocking the screen will also occur. After the 48-
hour closure, we noticed that the pump intake was also experiencing some clogging with the
plant debris from the aquatic plants in the dwelling tank. To combat this, another filtration
system was designed similar to the overflow screen in the bioreactor tank. It can be seen in
Figure 11. It extends to the bottom of the tank to the acrylic sheet on top of the tank. The bottom
of the filter is sewn shut with wire, so nothing can enter from the underside. The top sticks far
enough out of the water and is pressed against the acrylic sheet, so we are not concerned with
anything entering through there.
Figure 11. The new filter around the pump allows water to pass through, but not plant debris or
crayfish.

Sustainability considerations
We believe that our tank, although having a higher biomass and volume than the other
designs, is a more sustainable and cost-effective design. Our tank is very simple to build and is
built with cheap materials. The most expensive aspect of the tank is the aquarium tank itself, and
after that the light is the second most expensive aspect. The total cost was almost half of the
given budget and it sustained two crayfish weighing in at 50.7 g at the close of our testing.
Another sustainable aspect of this design is that many of the materials used were recycled from
old projects or locally collected. Since we recycled materials from older projects, recreating this
design would be more costly if all of the materials had to be purchased. We still believe that is is
a very affordable design and could be built for approximately $50 (the budget) worth of
materials if you already have an aquarium tank and light.
Conclusions and Recommendations
We believe our system was a success. We wish it could have supported more crayfish
than just two, but two is a good start and shows that our design functions. To improve our design
to support more life, we think that adding shredded PVC to the entire bottom of the tank would
significantly reduce the territorial behavior we witnessed when adding in more than two crayfish
to the tank. The shredded PVC method was proven in the cooler container the crayfish were kept
in while we built our microcosm. Another design we had in mind was to section off the aquarium
tank into cells to separate the crayfish from each other. This design would hold more crayfish
than our final design, but we believe the crayfish would not be as happy in this type of
environment. They would not have space to swim or crawl around if this design was built in a
10-gal aquarium tank, but if we used a larger tank, this design would be feasible. We still think
that given the space it would take to complete this cell design, that the shredded PVC would
work just as well and be cheaper to build. It would also be hard to implement the second level
above water section in the cell design. The downfall to the shredded PVC when compared to our
final design and the cell design is that it would be harder to locate the crayfish and see if they
were still alive when they are buried in PVC. The shredded PVC would also allow more algae
growth from the added surface area, which could then be eaten by the crayfish.
The two tank system worked wonderfully. This method is a proven sustainability tactic
and used around the globe in aquaponics. If we were building this on a larger scale, the
bioreactor plants could be switched to edible plants so that the plants could continue to clean the
water for the crayfish, but then the plants could be harvested along with some of the crayfish to
be eaten. This design would then become a self-sustaining farm. Our design proved to
sufficiently provide nutrients for our plants in the bioreactor as they looked unhealthy when they
were sitting in another bucket while we finished sealing the hole in the bioreactor bucket but
were rejuvenated once implemented in our system after a few days. Our system also supported a
snail unknowingly in the bioreactor for the entire testing period. This shows the possibility of
having other organisms, such as snails, living in the bioreactor. If using this for a farming
method, the snails could also be harvested and eaten.
Appendices (include raw monitoring data, design sketches, calculations, etc.)

Figures 12, 13 and 14. Weights of the crayfish and hornwort used
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Huner, J. V. 2011. Procambarus clarkii (red swamp crayfish). CABI Invasive Species
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