Algae+'14-'15

Welcome to the Newton North '14-'15 algae team's Wiki. Our members are Junghyun Park, Natalie Ferry, Tharun Kannan, Orenna Brand, and Adlai Hess. In this space, we will include helpful links and pictures, as well as short writings on the progress of our project. We hope that these elements will aid future algae teams and inspire everyone to continue the research and get excited about the small green organisms known as algae.

__November__ The month of November was primarily spent designing and constructing the system in which we plan to grow algae. Inspired by an Instructables design, Natalie and Orenna built a small bioreactor prototype, made simply of a 10-gallon fish tank, a small pump, and a few meters of tubing. The bioreactor is lit by a few compact fluorescent bulbs and a tubular shop light and is surrounded by reflective material to maximize light. While the girls focused on the bioreactor, Junghyun, Tharun, and Adlai did some research into species of algae. The group placed an order for 50 ml of chlorella and 50 ml of ankistrodesmus from our friends in Connecticut. In addition, they experimented with some leftover algae and algae paste from last year's team.



After an order number mix-up and a couple weeks of waiting, we received our shipment of algae. The package came just at the end of C-block, and, because it was perishable, Orenna and Adlai had to miss D-block and put the algae into the system. They put 40 ml of chlorella and 108 drops (6 drops per liter) of Miracle Gro into the bioreactor, the medium of which was DI water. The remaining 10 ml of chlorella and 50 ml of ankistrodesmus were put into a smaller water-bottle system meant to grow algae for the bioreactor.

Note: When we say bioreactor, we are referring to a system of tubing and lighting to optimize growth, NOT an actual bioreactor.

We then left for Thanksgiving break and stuffed ourselves with turkey.

__December__ We came back from break very excited to see what algal growth there had been, and we were thoroughly confused to see that the water in the bioreactor had turned a milky white color. The only green growth seemed to be at the bottom of the tank, and it was very minimal. In addition, about half a gallon of the medium had evaporated. In the small water-bottle system, much of the Universal Algae Medium had similarly evaporated, but we thankfully saw some more concentrated green colors.

Concerned by what we saw, we spent some time doing research on water turbidity and algal by-products. The group got in contact with Charles Mouchtaris (bioreactor designer) of Greece, who we hoped would be able to provide us with some insight. In an attempt to better the system, small changes were made to the bioreactor. The tank was covered with an acrylic sheet so as to lessen evaporation, and the lights and pump were put on a 12-12 cycle, in the hopes that the water might be able to settle overnight.



Over the next couple of days, we saw some off-white growth on the walls of the tank. It did not resemble algae at all, so we took a sample of it and put it under the microscope. We came to the conclusion that the algae was infected and had to be removed so as not to infect other cultures.



We finally had our long awaited Skype call with Charles Mouchtaris. He was able to provide a lot of insight regarding our lack of growth. We learned about ratio, air circulation, and light intake. The infection remains a mystery, but we have accepted defeat and are going to grow an entirely new culture with Charles's advice. Now, we just have to clean the infected tank.

__January__ We returned to the project after a wonderful holiday. Before leaving for vacation, we had emptied and cleaned the tank, leaving the tube and the pump out to dry. Natalie and Orenna reassembled the bioreactor in preparation for the second attempt at growth. The new bioreactor was made with adjustments based on Mouchtaris's advice: fewer and farther placed fluorescent lighting and an acrylic cover for the tank to lessen evaporation. The group had also decided on a 1/10 ratio (chlorella to DI water) for optimal growth in the bioreactor.

The boys got to work cultivating the algae. Under a rig of reflective surfaces and shop lights, they set up mason jars of chlorella and ankistrodesmus. Over a period of three weeks, they fed and diluted the aglae until reaching about half a gallon at ideal density, which was put into the bioreactor.

On the side, Orenna continued to work on her hydroponic farm, replacing the dying dill with new basil and sage plants. The roots of the original cilantro and basil have grown to become quite long, and the leaves have become unusually large.





__February__ Down by one member for the month, the algae team continued to fight to save the world. Although Orenna was in France, she didn't miss much because there were six snow days which drastically reduced the group's time in the lab. The direction and general goals of the project had to be readjusted to compensate for the lost time.

Unfortunately, the adjustments made to the bioreactor failed to work. When healthy algae was placed into the system, it lost color and died within two or three weeks, presumably because of the lack of motion in the water. And so, the focus has been taken off of growing algae in an efficient manner and turned to harvesting and extraction. All algae production has been moved into a new system of water jugs, bottles, and air stones. It seems to be thriving in the system, though we have lost the aspect of using natural sunlight.

In preparing to harvest, we have been cleaning out an industrial centrifuge which has not been used for some time and is full of green, vomit-smelling liquid. While the centrifuge does seem like a good mechanical option, it is not very environmentally friendly or efficient, so we are also considering chemical techniques such as the use of iron powder (an explanation on that to come later).

In Orenna's absence, the basil and cilantro from the hydroponic farm were moved into an aquaponic system where they have unfortunately lost a great deal of vitality because of the difference in nutrients in the water.

__March__

With a limited supply of live algae remaining, the group proceeded to experiment with different harvesting techniques. We considered the use of the centrifuge, but it would be very slow and take an incredibly amount of effort for an uncertain yield, so we turned our energy to chemical techniques. Using iron filings given to us by the chemistry department, we were able to experiment with iron-induced flocculation. When iron is mixed with algae, it forms iron oxide and sticks to the algae cell wall. This dense substance sinks to the bottom of the growth chamber (a water jug, in our case), allowing it to be separated from the water. We experimented with magnets and filters and found that it was very difficult to separate the iron powder from the algae.

A more successful means of harvesting, we found, is auto-flocculation. This technique is done by simply stymieing the algae's supply of carbon dioxide, a necessary ingredient for the algae's photosynthesis project. We did this by sealing the top of the growth chamber with duct tape. The algae went into 'survival-mode,' overcompensating for the lack of carbon dioxide by turning all of its carbohydrates into lipids. As the density increased, the algae sank to the bottom of the jug. We separated the dense algae from the remaining liquid by pouring the liquid out.

In addition, we experimented with potassium hydroxide, adding a 1 mol/L solution to a sample of algae. We observed that the algae, shocked by the sudden change in pH, clumped together and sank to the bottom of the growth chamber.

With the dense algae that we produced via these harvesting techniques, we made algae flakes. This form of algae is very commonly used in commercial production of algae-derived biofuels. We dried our 'algae paste' under heat lamps and collected the flakes, which were to be used in later steps of the process.

We took the resulting flakes and mixed them with a variety of organic solvents, primarily methanol and hexane. This yielded results that we will test into April.

__April__

This month began with preparations to wrap up the project, the first of which was a small exhibit in the Newton Free Library, thanks to the Green Decade Coalition of Newton. Each core team was represented by a small model or illustration of its system. For example, the algae team represented the algae-to-oil process with samples of algae at different points in the process. The exhibit included an brief explanation of the project.

We had to reexamine our long-term goals as the end of the project timeline came into sight: April vacation. We had originally planned to produce a large quantity of algae oil and turn it into biodiesel. Having grown a great amount of algae in an efficient system, we were satisfied with our progress thus far, and decided that our new goal would be to document everything so that next year's students may be able to continue our research. Documents containing the details of our research can be found on Google Docs.

Since dumping the remaining algae into the sink and shutting down the system, we have begun to brainstorm for our science inquiry and design project that will take us to the end of the year.

Here is some advice for next year:

Unless it is your goal, we recommend you skip the algae growing process and simply move to extraction with bought product. However, if you do choose to grow algae, here are some tips:
 * Stick with Miracle Gro, as it is a reliable growth formula and cannot go wrong. You may find the F/2 formula, but it is not worth pursuing and is difficult to procure.
 * It is better to slightly overfeed the algae than underfeed it.
 * More light is not necessarily a good thing.
 * Keeping a contained and (as much as possible) sterile environment is critical.
 * Try not to grow more algae than you need, or you may have to face the rather unpleasant job of disposal.
 * The centrifuge will likely be more troublesome than helpful. Instead, use potassium hydroxide to flocculate the algae and dump excess water from the upper portion of the mixture, then use a coffee filter to get paste for flakes.

Additionally, you may want to consider these general pieces of advice
 * Don't sit around idle; there are always things to do, be it research or cleanup.
 * Try to revive the mycelium research (should you be interested). If you attempt to do so, use this: [|MyceliumCultivationProcedure (1).pdf]

An important page:

http://www.fao.org/docrep/003/w3732e/w3732e06.htm

We hope that this blog has helped you and your team begin the year.

Sincerely, Orenna, Tharun, Natalie, Junghyun, and Adlai

Check out the previous year's work here: Algae '13-'14