Problem: Algae has proved to be a potential sustainable and eco-friendly energy source. The issue is, it grows too slowly and uses too much water. After doing some online research, we gathered information proving that adding sugar to the water when growing certain plants can lower water requirements in plant growth and enhance the growth rate because sugar gives the plants a direct energy source so they don't have to rely on photosynthesis for energy. It can also help maintain osmotic pressure in the plants' cells, reducing the amount of water required to grow, and it contains carbon dioxide, so by giving the plants an external carbon source, the plants will grow at an enhanced rate.

Hypothesis: If we add sugar to the spirulina algae, then it will grow faster and require less water.

What is microaglae?

Microalgae falls under the family of a plant known as algae. It is a single - celled organism known mainyl for its capabilities of not only being a good food supplement but also for its ability to quickly photosynthesise, which has lead to its success for the useage of biomass to create into biofuel. They are also known for their large contirbution of oxygen into aquatic enviorments where they are grown. 

What is biomass and how is it created? 

Oganic matter which is used as fuel, to create biomass fuel there are numerous different options such as combustion, anaerobic digestion (produces methane), fermentation (produces ethanol), and pryolysis( produces bio-oil). In this study we will be focusing on fermentation but will also touch on pryolysis. This is due to the fact that these two are more correalated to the useage of microalgae and most research is based off of these two methods. A part from this biomass in general is a renewable soruce of energy that is a new source of research as a substitute for the fossil fuels that make up the primary aspect of our energy consumption currently. However all new sources of energy have their issues especially when their potential and issues have not been fully investigated, with our study our aim is to improve this new growing sector of energy for the future of all renewables. 

What are some of the biggest issues with biomasss as a form of electricity? 

Uses 70 to 400 times more water than any other form of energy and the time used to grow the algae needed to create biomass and biofuel is very ineffiecent and timely. With the algae itself taking a total of 4 weeks to be able to harvest on average and to develop growth, this time period excludes that it then takes to purify the biomass to create the biofuel needed in ethanol. However this is still at a high increase compared to that it would take if the material being used was the traditional wood that many companies use that takes several weeks, and takes a much higher toll on the enviorment removing the very trees that we need to get rid of carbon dioxide and monoxides from the enviorment. As a result algae as a form of biomass is more efficent, despite this the process still takes quite a bit of time. 

How to effectively grow algae: 

Find a section of your home where sunlight is constant for at least eight hours a day. Set up however many jars needed in relation to the amount of algae being grown. Make sure you have algae nutirents that are effective as well as nutrient salts that have not been in contact with water prior to beginning algage growth process. Ensure that algae growth jars are in an area where the temperature stays relatively constant and around 20-35°C. When creating algae setup, ensure that all ingridients are acurately measured and there is no spilage. While algae is growing, shake the jar(s) daily. Algae is ready to harvest in 2-3 weeks or when algae is a vibrant green or grey, and there are solids in the water or on the side of the jar.

Why is creating a way to grow algae for biofuel useful in our society?

With researching environmentally friendly ways of gathering energy are becoming more and more frequent, biofuel becomes a vital incorporation into 'envrionmentally friendly future' discussions. Biofuel is a way to sustainably utilize energy that fuels vehicles allowing humans to lower climate change rates and prevent global environmental issues. Since vehicles make up for 15% of global CO2 emmisions, switching to biofuel would significantly improve the Earth's current environmental state. Since algae can be a reliable source of biofuel, being able to find a way to produce is more efficiently and with less water becomes essential to becoming a more environmentally friendly society.

Conditions needed and how to culture:

In order to prepare a solution for the spirulina to be cultivated nutrients in order to form proteins and to help the general function of the cells, while salts are required in order for cultivate the culture. Using a chlorine filtered water in order to support the Spirulina the specific species that we will be cultivating in the experiment, using bottled filtered water as a result especially for consistent filtration in order to keep our experiment consistent. The culture also required white LEDS lights, this is what is typically used in the lab setting and as a result what would most likely be used in production, therefore we used this light during our expirement. 

Nutrients (chemical enviorment) containing: 

Vitamins

• Thiamine: Reduces the stress thiamine puts on the enviorment, specifically on other living organisms in the the enviroment. Allowing the specimen the ability to reproduce if damage is taken to tissues or cells specifically during the reproduction and replication processes. A critical component when applied to a spirulina biomnass study where the objective is to see the greatest amount of reproduction. Thiamine also promotes the release of a chemical known as salicyclic acid, this prevents abiotic and biotic stresses found in the enviorment, while also helping the cirtical components germination, senescence, abscission, fruit ripening. 
• Biotin: Produces fatty acid synthesis, energy production, along with other metabollic reactions. This can include the breakdown of these fatty acids enabling the spirulina to function, and have accesible access to energy production methods, also including processes such as gluconeogenesis and amino acid metabolism. Each of which aids in the obtaining and application of the useage of energy through the higher rate of production biotin enables an enyzme to function. 
• Cyanocobalamin: Works to help the growth of the plant and works to helps supplement needed vitamins. Working towards helping the spirulina build up resistance to changes in temperature, Ph and enviorment, all of these factors which are beneficial in a household enviorment. While also reinforcing the creation of enzymes to help breakdown proteins supporting energy production. 

Macronutrients

• Nitrate (sodium nitrate mainly): Safely manages the growth of the spirulina, this is through the limitation of the oxygen the culture can obtain. This brings safety into the home enviorment and keeps the algal bloom from surpassing the limits of the ecosystem or enviorment. Working as a fertilizer for the spirulina and boosting growth, this is through the process of supplying nitrogen to the spirulina which is needed to make proteins for the overall growth of the organism. 
• Phospate (usually sodium phosphate, phosphoric acid, or ammonium phosphate): Phosphates are one of the key components for algae growth through three main ideas energy transfer, the synthesis of DNA, and upholding membrane structure. Particularly through the compound adenosine triphosphate which is responsible for powering cellular activites, transfering the phosphate creating these energies. While the phosphate itself when combined with tetraoxide aide in cell division and reproductions. The upholding membrane structure derives from the fact that phosphate ensures that the cell functions properly and does not mutate conforming to its designated cell structure. 

Micronutrients 

• Iron: Facilitates the processes of photosynthesis and repsiration
• Copper sulfate: In small amounts helps the algae sustain itself through supplying the proteins neccessary for growth and repair of the tissues in the cell. While also supporting a process called the electron transport chain, which is known for its properties of photosynthesis. Assisting the conversion of sunlgiht into energy to produce sugar. 
• Molubdenum: The two main objectives of using molubdenum are processing DNA material and proteins. Helpign enhnace growth and in the process using a decrease in materials, aiding in the releasage of energy. All of these important aspeccts when bringing in how to best foster algae growth. 
• Zinc: The main function of zinc is to increase DNA synthesis, and enzyme activation. This works through zinc a tivating these ensymes allowing for increased metabolism of proteins, carbohydrates, and lipids, increasing the efficency of producing sugar and releasing energy. While also protecting against damages from compounds of oxygen such as hydrogen peroxide, this is through helping in production of an enzyme known as superoxide dismutase. Zinc also aids in data synthesis/ production increasing the reproduction while also minimizing mutations. 
• Cobalt: Promotes overall plant growth and health; faciliatting the relief of stress if it is changing of temperatures, nutrient scarcity, or changes in water conditions. Ensuring that the algae would not be as affected by changing conditions in the enviorment, meaning it can be used as a reliable supply. 
• Manganese: In small doses leads to enzyme activation, higher levels of metabolism, and photosynthesis. Manganese assists algae specifically in the area of splitting molecules and putting them back in the enviroment, coverting light ito energy known as chemical energy. While also helping produce an enzyme which breaks down lipids, carbohydrates, and proteins. While also monitoring chemicals such as superoixide, hydrogen peroixide, and hydroxyl radical which neutralize these chemicals. 
• Please note: This includes different compounds of these micronutrients

Overarching factors: 

• The main focus of these nturients is to bring stability to the Spirulina culture and increase the reproduction abilties of the Spirulina culture. 

How to asses Spirulina strain:

In order to confirm the success or failure of the experiment as a whole we delved into the topic of what the strain needed to look like. Having characterisitics that clearly defined the organism being known to have a spiral texture and sometimes have two of these intertwine also known as a double helix strucutre. While smaller samples are known for being smaller spots as they have not fully developed as of yet. Through this research we also discovered that to properly view the spirulina we needed to do a fault manuver to prepare the sample in order to trap the bioorganism and also the liquid used to cultivate the species, along with use a 40x magnification level. This is due to the fact that the organism itself is very miniscule not being able to be viewed if not at a 40x magnification level due to the size of their trichomes and being a single-celled organism. However a 60x mangification level is preferable if available. (was not available for our samples) 

Test 1: Can the favoured variable make the algae grow faster?

Control: Algae with the water and nutrient measurements according to package directions (1.5mL nutrients, 100mL cuture, 8g culture salts, 500mL water).

Variable 1 - increased nutrients: Algae with water measurements according to package directions and increased nutrients (5.6mL nutrients, 100mL cuture, 8g culture salts, 500mL water).

Variable 2 - added sugar (Favoured variable): Algae with the water and nutrient measurements according to package directions as well as dissolved sugar (1.5mL nutrients, 100mL cuture, 8g culture salts, 4.5tsp sugar, 500mL water).

Test 2: Can the favoured variable make the algae grow faster with reduced water measurements?

Control: Algae with nutrient measurements according to package directions and reduced water by 40% (1mL nutrients, 100mL cuture, 8g culture salts, 300mL water).

Variable 1 - increased nutrients: Algae with increased nutrients and reduced water by 40% (5.6mL nutrients, 100mL cuture, 8g culture salts, 300mL water).

Variable 2 - added sugar (Favoured variable): Algae with nutrient measurements according to package directions and reduced water by 40%, as well as dissolved sugar (1mL nutrients, 100mL cuture, 8g culture salts, 4.5tsp sugar, 300mL water).

Test 1: Can the favoured variable make the algae grow faster?

• Control: The control jar was created with 500mL of filtered water, 1.5mL of algae nutrients, and 8g of spirulina culture salts. The media was shaken well until turbid and then left to settle for twenty four hours in a dark, damp room to receive optimal results. It was then placed under a white LED light for the remainder of the three week test period to maximize energy efficiency without too much heat.
• Variable 1 - increased nutrients: The increased nutrients jar was created with 500mL of filtered water, 5.6mL of algae nutrients, and 8g of spirulina culture salts. The media was shaken well until turbid and then left to settle for twenty four hours in a dark, damp room to recieve optimal results. It was then placed under a white LED light for the remainder of the three week test period to maximize energy efficiency without too much heat.
• Variable 2 - added sugar (Favoured variable): The added sugar jar was created with 500mL of filtered water, 1.5mL of algae nutrients, 8g of spirulina culture salts, and a total 4.5tsp of dissolved sugar (1.5tsp added per week). The media was shaken well until turbid and then left to settle for twenty four hours in a dark, damp room to receive optimal results. It was then placed under a white LED light for the remainder of the three week test period to maximize energy efficiency without too much heat.

Test 2: Can the favoured variable make the algae grow faster with reduced water measurements?

• Control: The control jar was created with 300mL of filtered water (40% less than test 1), 1.5mL of algae nutrients, and 8g of spirulina culture salts. The media was shaken well until turbid and then left to settle for twenty four hours in a dark, damp room to receive optimal results. It was then placed under a white LED light for the remainder of the three week test period to maximize energy efficiency without too much heat.
• Variable 1 - increased nutrients: The increased nutrients jar was created with 300mL of filtered water (40% less than test 1), 5.6mL of algae nutrients, and 8g of spirulina culture salts. The media was shaken well until turbid and then left to settle for twenty four hours in a dark, damp room to receive optimal results. It was then placed under a white LED light for the remainder of the three week test period to maximize energy efficiency without too much heat.
• Variable 2 - added sugar (Favoured variable): The added sugar jar was created with 300mL of filtered water (40% less than test 1), 1.5mL of algae nutrients, 8g of spirulina culture salts, and a total 4.5tsp of dissolved sugar (1.5tsp added per week). The media was shaken well until turbid and then left to settle for twenty four hours in a dark, damp room to receive optimal results. It was then placed under a white LED light for the remainder of the three week test period to maximize energy efficiency without too much heat.

Test 1

Sugar:

• Addition of sugar caused oxygen to be released and bubbles to form on the surface, was able to be heard clearly 
• Performance largely out manuvered than that of the two other experiments. 
• Showed higher turbidity and growth rate during the span of the first day
• Turbidity developed on third day - cloudiness persisted consistently 
  • Likely result of the dissolvance of the sugar as commonly can create this variable 
  • Also a result of higher density of particles as seen throught the results of the optical density test 
  • Developed sour milk smell after 5 days (normally means the culture has reached its peak based off of resources) 

Nutrients:

• Performed second highest
• Optical density had a higher concentration than control but higher weight 
• Growth rate increased during the middle of the experimental process
• Particles remained suspended after mixing 

Control:

• Remained consistently clear, menas low particle level 
• Few particles dispersed throughout the liquid comparitively 

Test 2 (decreased water consumption)

Sugar:

• Addition of sugar caused oxygen to be released and for a reaction to take place that ultimately has led to a higher rate of growth
• Developed mainly during first day
  • Unlike test 1 developed ond ay 2 and 3 but then stopped past that point
  • Amount of culture grown was visibly larger 
  • Stuck to rim and floated on surface of water 
  • Had consistent base coverage

Nutrients:

• Developed a gaseous release when extra nutrients were added 
• More strand growth compared to test 1 
• Particles floating on surface instead of bottom 

Control:

• Little to no progress over total course of experiment, devleoped minimal culture 
• Culture sank to bottom, most of development was on the jar bottom not on the surface  

Please note this is summarised version as most of detailed observations can be found in the logbook 

Microscope testing results: For First Testing Batch

The strain viewed through all three samples was consistent in the type of organism that was seen. This means that there was no cross contamination of the organisms themselves. We first harvested the algae which was done through using a fine strainer to separate the liquid from the algae itself. We prepared the sample using a clean pipette each time we dropped two drops of the concentrated sample onto the slip. We then, using a 45 degree angle, dropped the coverslip in order to prepare all 3 samples. We then used the magnification of 40x for each sample while keeping the elevations consistent throughout the experiment. The result was we found that the sugar strain of the spirulina the one showed to the right had the longest sample, meaning the organism developed the most in this situation. Having the highest rate of success, while the nutrient sample  performed similarly to the standard sample, with both not developing the strands seen in the sugar sample. However we can infer that the nutrient sample was successful at a higher rate due to the fact that there was more volume of the substance itself when harvested. 

Microscope testing results: Second Testing Batch ( decreased water consumption ) :

Out of the three strains viewed the sugar by far performed the best. Having developed the largest amount fo culture, as seen through the samples attached below. The organism found throughout hte three samples also stayed consitent meaning that none of the samples were contaminated. When it comes to the preparation of the sample for analysis we dropped two drops of the concentrated sample onto the slip. We then, using a 45 degree angle, dropped the coverslip in order to prepare all 3 samples. We then used the magnification of 40x for each sample while keeping the elevations consistent throughout the experiment. Using a different pirepette each time to prepare the sample for observation. With the sugar sample we would also like to note that the sample remained consistent since day one, with little further growth. Despite this the nutrient and control were very far behind with the both enedwing up having similar results.

Progress brief overview/ summary: 

• Sugar:
  • Performed the best, most culture grown 
  • Lowest optical density (meaning highest concentration ) and lowest weight ( refering to the amount of water absorbed by culture) 
• Nutrients 
  • Performed adequately in comparison to the contol, having a higher performance however it lacked the ability to completely overtake the control variable as seen with the progress of the sugar 
  • Had a comparitive optical density to control 
• Control 
  • Lowest performace out of all the variables being the control, it had a high optical density (lowest concentration of particles)

What did we learn from this experiment and what did we accomplish?

Overall, we learned a lot about growing algae in this process and are more determined than ever to help others see the possibilities with algae as biofuel. Our hypothesis was assuming if we add sugar to the spirulina algae, then it would grow faster and require less water. Throughout our experimentation process, we discovered that incorporating sugar into the nutrient bases for growing algae was a fantastic way to get the results that we wanted. The sugar gave the algae excess energy accelerating its growth time and making it grow at a rate suitable for biofuel. We also concurred that sugar assisted in maintaining the algae's osmotic balance in the cells, allowing the algae to retain water and function efficiently despite there being less water. This experimentation process was both intriguing and groundbreaking because we were able to solve two main issues with using this algae as biofuel. This means that within the next thirty years, algae production can start being used as sustainable energy to reduce greenhouse gas emissions and climate change, resulting in a greener planet and a healthier environment.

The innovation of the spirulina with the result of expediting growth applies directly to the field of electricity. With this new advancement which has led to the more efficient processing of the material (spirulina), this will cause there to be a higher usage of biomass in the future. Being a new resource of electricity, the innovation in this area lets us better understand how this form of electricity functions and the new innovations that can be made in the future. The usage of our research will lead to innovations in the field along with future ones as now, one of the largest problems in the industry is being addressed. 

•  Not testing multiple samples 
  • Although intended to test Spirulina s the main algae we still have to take into account the multiple different types such as Chlorella as success can vary as a result, and the success rate of the solutions. 

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We would just like to extend a thank you not only to our parents who have supported our endeavors but also our teacher Mrs. Calvert, who has taken away from her own time to help us gain access to resources and information. To everyone else who has helped us get to where we are we are thankful for all you've done and assisted us with.