Agrisept-L Testing as an Algeacide

[bobbyo]

Hello, This is my second letter, not sure if the first went through. I would like to test the fungicidal qualities of agrisept-l. I would purchase live algae then run an experiment testing the algae growth vs the quantified addition of agrispet-l. Does this sound like an experiment that is appropriate for 10th grade. I hope so because it seems very interesting to me. Bobbyo

[Snobs3747]

Sounds interesting to me too!
I'd like to let you know that if you find something interesting, you should definitely pursue it and research it further. There is no "10th-grade appropriate" type of science fair project. The best projects are the ones that are extremely well-done and interesting. Also, I have never seen this project before in any science fair I've been to, so it seems unique enough to me!
If you still don't think it is "appropriate", you can keep the same concept/materials and expand on it. You could do a project relating to algae growth, but one that tests a more complex hypothesis.
Let me know if that helps!

[heatherL]

Hi Bobbyo,

You have a very interesting and feasible project idea! I just wanted to clarify something for you: algae are not fungi. If you want to test the fungicidal properties of Agrisept-L, which appears to be used to kill all kinds of microbes, you would need to test it with a fungus, like yeast. Yeast is easy to grow on petri dishes, and you could definitely do a project with yeast. That said, it is perfectly interesting to test whether Agrisept-L kills algae; just make sure you do not refer to algae as fungi.

Please post again if you have more questions.

Heather

[donnahardy2]

Hi Bobbyo,

Snobs3747 and heatherL have give you some really good ideas for getting started on this very interesting and challenging project. I have some additional suggestions:

There are 3 project ideas on the Science Buddies website that describe experiments for measuring the antimicrobial effects of agents on various microbes. Although none of these projects describe how to grow algae, they should give you inspiration for designing your own experiment. Please note that it is important to test the dose response when doing this type of testing.

Hhttp://www.sciencebuddies.org/science- ... p014.shtml

http://www.sciencebuddies.org/science-f ... p013.shtml

http://www.sciencebuddies.org/science-f ... p016.shtml

There is one project idea that involves testing the effect of fertilizers on algae growth on this website and the experimental set-up might work for your project:

http://www.sciencebuddies.org/science-f ... p054.shtml

Here is a summary of a project on algaecides that should give you useful background information and an idea of how to do an experiment in a larger body of water:

http://www.pacificorp.com/content/dam/p ... tStudy.pdf

You should also do an internet search and look for more information on algae and algaecides. Your goal should be to become an expert on your topic.

I have a couple of questions also. What type of algae are you going to use? And, what will you use for a control?

Donna Hardy

[bobbyo]

Thank you so much for replyging. I will be testing algae soon sometime in November. I will get back to you when things more evolved. Thanks again!

[donnahardy2]

Hi,

Thanks for your reply. It sounds like you are making progress.

At this point, you should be defining your hypothesis , and designing a carefully controlled experiment. Be sure to review the guidelines on the Science Buddies website for a hypothesis and designing an experiment before you start.

http://www.sciencebuddies.org/science-f ... ndex.shtml

Do try to plan ahead so you will have time to do your experiment twice.

Donna Hardy

[bobbyo]

Hi Again,
After further investigation, I have decided to test agricept-l and its proposed aglaecide quality. I will be using the algae species ankistrodesmus. I can obtain the algae and pipet 1ml quantities into glass test tubes. I will use a fluorescent lighting. I think the best way to quantify algae is to gauge the "greeness" from daarkest green to the least green. My question is what percent solutions of the agricept should I use? I was thinking 1%-5% solutions. But should I make it more of a 5%, 10%, on up to 50% solution. I'm concern oof the cost factor of this testing. Also how many times should I repeat each test, 5 times, 10 times, etc. I need to know before ordering my algae. Thanks.

[donnahardy2]

Hi,

It's good that you are finalizing the details of your experiment.

Do you have access to a spectrophotometer so you can measure the density of the green color at 550 nm? This would give you a quantitative measurement of your results. Or, do you have access to a microscope? I believe you could also count cells in a hemocytometer. However, measuring absorbance is much easier.

Here's a website that describes various methods for measuring algae:

http://www.botany.wisc.edu/courses/bota ... ations.pdf

Do you have time to order a small culture of the algae and grow it in a stock container so you will have fresh and rapidly growing algae to start your experiment? Results for experiments like this depend on having cells that are at the same phase of growth, and algae that you receive by mail may take a while to recover and start growing. And you would only have to pay for the initial small culture.

What concentration of algaecide is recommended on the product label? You want to test a range of concentrations, perhaps 5-6 concentrations, that are below, at, and above the recommended concentration. Also, you need a control with no algaecide.

I recommend doing a pilot experiment with a control and one concentration of algaecide to test your method and find out about any experimental problems you will encounter. Then plan to do your experiment in duplicate, or better, triplicate. Using a higher number of replicates will allow you to do statistical analysis of the results, so more replicates would be better. However, each sample will require a lot of work.

Donna Hardy

[bobbyo]

Hello Again,
I have doing a lot of research and have made a hypothesis. I will be testing agrisept and its individual ingredients for their algaecidal properties. Agrisept is made up of grapefruit, tangerine, lemon, and lime seed extracts. I could buy the agrisept and the extracts. I was wondering about the extracts. when you purchase the extracts, they are in a glycerine medium. I could make my own by grinding the seed, non juicy pulp and whites of the fruits and just add them to a basic spring water. they can be quantified and the spring water is best for growing my green algae. Would you recommend buying the seeds extracts or making them myself? I know have cellometers and a great microscope. My hypotheses is that the agrisept will be 4x algaecidal then the single fruit seeds. Then grapefruit will then be 3x greater then the other. Following lemon, lime, then tangerine. This is based on research. I will be dealing wiTh serial dilutions. I will contact you more about that! Algae will be arriving in two weeks. Wanted to make sure everything is ready. Thanks-Bobby

[donnahardy2]

Hi Bobbyo,

It sounds like you are making great progress. You can either purchase the individual extracts or prepare them yourself. Either would be scientifically valid as long as you explain exactly what you did for your experiment.

If you prepare the extracts yourself, you would want to make sure you have some way of measuring the extract so that anyone reading your project would be able to replicate your results.

If you purchase the extract, I would recommend checking with the supplier first and find out what they do to test the product to ensure reproducibility from lot to lot. Ask if you can obtain a certificate of analysis for the batches that your purchase.

If you can’t obtain any information at all from the manufacturer, then you might decide it would be best to make your own. If the manufacturer can supply certificates or other helpful information, then you might decide to purchase the extracts.

Testing a series of dilutions for each sample is a great idea. Are you going to be measuring algae growth using a hemocytometer?

Please post again and let me know about your progress.


Donna Hardy

[bobbyo]

Hey again, I contacted the distributor and asked suggested questions. Awaiting reply. I have cellometers which are like the hemocytometers to do my counts. I have to more questions. 1. Is this a strong hypotheseis? If ankistrodesmus is introduced to varing concentrations of Agrisept or it's individual componets, then the algae will die quicker with the Agrisept than each individual component. 2. I listed myself under botony, should I be under biochemistry? Thanks!

[donnahardy2]

Hi Bobbyo,

Your hypothesis is very good and you have the equipment to obtain quantitative results. This is definitely a botany project, but it could also be entered under biochemistry.

Good luck! Let us know what happens.


Donna Hardy

[bobbyo]

Dear Miss Hardy,
I am a little frustrated. I am ready to go but I haven't had any responses from the people I have contact( 2 separate places) for questions you suggested asking from the distributor. I did follow up phone calls but still haven't has responses. I don't want to wait too long because I need to start experimentation .
I do have 2 question regarding my serial dilutions. I know how to serial dilute my algaecide from 1:10 to 1:1000. I plan to introduce a quantity of algae to these dilutions separately. I then plan to centrifuge my algae/diluted algaecide to see how may live alge cells I have. I will be using a cellometer that requires a 20 ul amount of suspension.
1. Should I centrifuge( hand made hand-held centrifuge) or just mix algae with diluted algaecIdes?
2. The cellometer instructs to multiply the counts by 10 to 4th power than multiply by dilution factor. Does that mean if I count 50 cells at the 1:10 dilution of agrisept X 10 to 4th = 5,000,000 cells per ml? I think I'm missing something about the quantity of algae I add to the algaecide dilution. Where does that come into play. For instance, if I pipette 100 ul of algae into all the different dilutions of algaecides do I times that somewhere in the calculations? I'm getting confused- think I need a break!!! Thanks Bobby

[donnahardy2]

Hi Bobbyo,

I’m sorry you are not getting the information you need. Some companies are good about answering questions, and some are not. I would go ahead and start your experiment and not wait for additional information.

If you have time, you could try calling the company’s toll free number and explain why you need the information.

It would be good to get a count of the algae before you introduce the algaecides. This will be your time zero count. Then add the algaecide to the samples at time zero. The concentration of algae at the beginning of the experiment is one of your controlled parameters.

You are correct! If you make a 1:10 dilution, then you would multiply the number of cells counted by 10 to the 4th x 10. So if you count 50 cells, the number of cells would be 5,000,000 per mL.

Donna Hardy

[bobbyo]

Thanks for all your support. Today I have a company who was very willing to send me a certificate of analysis with the items I purchased. I will ask my algae specialist at Carolina Biological more about how much ankistrodesmus would be good to introduce to each dilution of algaecides and approximate time I should wait ...I figure a half hour at first. This may be a trial and error pretesting run to figure times and amounts. All will be logged in notebook.

Would my controls be considered the algae in its spring water medium at a 1:10,1:100, 1:1000? And at o time read for each run and than the wait times (maybe 30minutes)? A control should be read along with the material be tested right?

Ready to run. Bobby O