lakshu_s
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Super-absorbent polymer prepared from orange peels

Postby lakshu_s » Tue Jul 23, 2019 11:07 am

Dear Mr. Willey,

It was a awesome experience working with you last year for my 8th grade project. Now as I step into 9th grade, I realized there are 2 categories of competitions one at regional level and other for Honors. I would like to take my project for Honors as well. But first let me analyse further into the topic I have chosen so I can prepare some paper work for pre-approval.

The project I have decided for this year is : Biofixation for CO2 using Coccolithophorid Algae.

First let me go step by step with my understanding. The abstract of the project is as below:

This project explores the CO2 fixating abilities of coccolithophorid algae. Coccolithophorid algae have the ability to produce external calcified cells known as coccoliths. These coccoliths require CO2 in order to be produced. The advantage of this is that unlike most algae (or trees) which release CO2 when they die, these algae keep it locked up in their shells, which sink to the bottom of the ocean and produce limestone. This means this CO2 will permanently be locked up and not released back into the atmosphere.


In this experiment the algae were put under different concentrations of Fe (NO3)3, Iron (III) Nitrate. The different concentrations the algae are tested u. The best iron concentration will be determined by testing the amount of CO2 left inside the bottles after 3 and 6 days. The gas will be tested with a CO2 gas censor. Doing this will arrive at a close number to what the algae can tolerate in terms of iron and which is the best for their growth.

My questions for my initial understanding are:

1. Where do I get Coccolithophorid Algae for experimenting
2. Why do we put the algae under different Iron Nitrate concentrations
3. How do I get Iron Nitrate and how do I measure the concentrations for testing
4. How do I test the amount of CO2 inside bottle (CO2 gas sensor)

All these are new terms to me but this interested my research. If you could help me with understanding I can proceed further.

Thanks,
Lakshita


[Title of thread changed 10/24/19 from "Help Needed: Experiment with Coccolithophorid Algae"]

LeungWilley
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Re: Help Needed: Experiment with Coccolithophorid Algae

Postby LeungWilley » Wed Jul 24, 2019 6:30 pm

HI Lakshita,
I am very glad to hear that you had a great summer and, reading your post below, it sounds you are ready to go for 9th grade!

Unfortunately, I am very sorry to say that this particular topic is way outside my area of expertise. Our moderator is reaching out to other experts who will be able to give you better feedback on this subject.

I wish you the best of luck in 9th grade and much success with all of your science experiment!
Willey

lakshu_s
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Re: Help Needed: Experiment with Coccolithophorid Algae

Postby lakshu_s » Sat Jul 27, 2019 10:17 am

Dear Mr.Willey,

Thanks for your wishes. It was really a good experience working with you last year. I am looking forward to get support from other expert who could guide me for this project. Already I have so many questions in my mind :(

Thanks,
Lakshita

LeungWilley
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Re: Help Needed: Experiment with Coccolithophorid Algae

Postby LeungWilley » Sun Jul 28, 2019 4:55 am

Hi Lakshita,
Likewise, it was great working with you last year as well!

By the way, one of our moderator, MadelineB, found this website of a experiment that's similar to what you have described. You might want to take a look:
http://www.odec.ca/projects/2008/sidd8g2/index.html

The site also provided some location for obtaining the Algae as well:
https://www.wardsci.com/store/product/8870194/coccolithophora

Good Luck!
Willey

lakshu_s
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Re: Help Needed: Experiment with Coccolithophorid Algae

Postby lakshu_s » Mon Jul 29, 2019 10:29 am

Thanks so much Mr.Willey.

Yes project description you specified is exactly the same. But I have so many questions in my mind to proceed with this project as so many terms are new to me.

Please let me know if I can ask you or please suggest me someone who can help me with those questions. After my initial questions are answered I can proceed step by step with the project.

Thanks,
Lakshita

lakshu_s
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Joined: Fri Aug 26, 2016 11:48 am
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Re: Help Needed: Experiment with Coccolithophorid Algae

Postby lakshu_s » Mon Jul 29, 2019 10:56 am

Also I found Iron Nitrate in the same website. Guess we should purchase : 470301-404 IRON (III) NITRATE 9-Hydrate Crystals Laboratory 100G. The procedural steps are confusing and I am still going over. It would be really great if I get some assistance in understanding the procedural steps. I see many of the materials are new to me :(

Thanks,
Lakshita

LeungWilley
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Re: Help Needed: Experiment with Coccolithophorid Algae

Postby LeungWilley » Mon Jul 29, 2019 6:27 pm

Hi Lakshita,
Please feel free to go ahead and post your questions on this forum. I may not be able to answer all of them and other experts will likely need to jump in / help clarify some of the more advance concepts.

At any rate, if there's a particular section of a procedure that's confusing, please let us know.
Thanks!
Willey

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Re: Help Needed: Experiment with Coccolithophorid Algae

Postby lakshu_s » Fri Aug 02, 2019 9:08 am

Dear Mr.Willey,

How are you. Thanks for willing to help me out. I am trying to understand and analyzing most of the new terms :( before I post my questions. Lot are new to me. I will send my understanding part by part and post my questions by this weekend.

Thanks,
Lakshita

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Re: Help Needed: Experiment with Coccolithophorid Algae

Postby lakshu_s » Fri Aug 02, 2019 2:51 pm

Dear Mr. Willey,

I got the basic understanding of what we are trying to achieve and the importance of this project.
First let me identify the materials needed for culturing the algae. Can you please help me with finding the materials at the right place.

1. As recommended by Ms.Madeline I went to ward science website for algae.
https://www.wardsci.com/store/product/8 ... lithophora

My question: Can we get this kit a month before starting the actual experiment? So when ever we want to culture the algae we can open this and start culturing right? Is there any precautions and storing I have to keep in mind when not using. Why I am asking this question is, in the attached PDF mentioned in the website under the topic "Maintaining Algal Cultures" in page 3, they asked to loosen the jar cover and store in cool area.
Hope that is not required in our case since we are going to do culture by separate method. Attaching the PDF herewith.
Please help me understand how to handle the package once received in order to prevent any contamination.


2. Ferric Nitrate: I saw this chemical in the same website. Hope we have to get Iron(III) Nitrate Nonahydrate
https://www.wardsci.com/store/catalog/p ... 470301-412
But seems I am not able to order even though my mom registered in this website. Can you please guide on this.
Guess I need to order crystals only right so I can prepare the solution as below.

In the project procedure to prepare Fe (NO3)3 Solution below are the steps:
1. Weigh out 10.30 grams of Fe (NO3)3.
2. Pour 250 ml distilled water into beaker.
3. Add the 10.3 grams of Fe (NO3)3 into the distilled water.
4. Mix until completely dissolved.
Each ml of this solution will give 0.024 ppm dilution in 250 ml water.
My question: Can you please explain me how each ml will give 0.024 ppm dilution ? can you please help me understand this calculation as we did for last project.

3. Tubing A will refer to any tubing with ¼” OUTSIDE diameter
I found the below in Home depot. Is it correct or do you suggest anyother tube specific to this project
1/4 in. O.D. x 1/6 in. I.D. x 25 ft. Polyethylene Tubing - home depot - Tube A

4. Tubing B will refer to any tubing with ¼’ INSIDE diameter.
I found this also in Home depot. If any suggestion please let me know.
3/8 in. O.D. x 1/4 in. I.D. x 10 ft. Clear PVC Vinyl Tubing B - home depot

My question: We have to insert tube A into tube B. Please let me know if what I am ordering is correct to satisfy this requirement.

5. Can you help me find in-line air filter, air pump, 1/4 inch elbow hose fitting, multi-port manifold (4 port manifold)
I found 1/4 inches Elbow hose fitting and mulit port manifold in home depot
1/4 in. Barb Elbows (10-Pack)
4-Port Manifold with Adjustable Flow
Not sure if this is right one.

6. I also see ¼” nut in the materials list. Do we need this?

I found this also in home depot.
Everbilt 1/4 in.-20 TPI Zinc-Plated Hex Nut (100-Piece per Box)
Model# 801730

My question: Below is where I see using the nuts. Can we go ahead without nut? Hope this is the tighten the port right.
This is the project steps specified:
Attach one end onto the in line air filter.
Attach the other end into the locking part of the ¼” elbow hose fitting.
Attach the other end into the multi-port manifold, the side with one port.
Plug one of the 4 ports on the opposite side with ¼” nut.
Attach 3 elbow hose fittings in the remaining ports.
Attach 90 cm piece of tubing A to one of the elbow hose fittings.

7. I am planning to do the entire project at home. Hope this should be fine. Can I hire microscope from any websites as it would be really costly?

I will wait for your suggestions in all these. Meanwhile I will continue researching the steps.

Thanks again for your time explaining me understand each and every step while I proceed with the project. Sorry as I mentioned I have not been introduced to so many terms mentioned in this project.

Thanks and have a nice weekend.
Lakshita
Attachments
Working_with_Algae_and_Cyanobacteria.pdf
(54.39 KiB) Downloaded 104 times

lakshu_s
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Re: Help Needed: Experiment with Coccolithophorid Algae

Postby lakshu_s » Tue Aug 06, 2019 10:20 am

Dear Mr.Willey,

How are you. If you can please reply to my previous email atleast portion by portion, I can start preparing for that and arrange to buy stuff.

Thanks,
Lakshita

LeungWilley
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Re: Help Needed: Experiment with Coccolithophorid Algae

Postby LeungWilley » Tue Aug 06, 2019 6:38 pm

Hi Lakshita,
Sorry for the delayed response. These are excellent questions:

Like you suggested, I am going to respond in parts:
1. You are on the right track and the information that you are looking for is in the second PDF titled: Algae & Cyanobacteria.
https://www.wardsci.com/assetsvc/asset/en_US/id/16920341/contents. From the document (under the AVAILABILITY heading) "In its packing container, the culture can retain its high quality for 4–7 days at room temperature. If the culture is not going to be used in this time frame, it should be subcultured to prevent death and overcrowding." (Also, please make sure you go to the last part of the document so that you have a plan to properly dispose of the algae when you are done with the experiment). Based on this and your concern about contamination, I would recommend that you wait until about a week before you are ready to perform the experiment to get this.

2a. Based on what I am seeing, Ferric Nitrate is sold and shipped to schools and businesses only. (Not available to individuals or residential addresses). Is there any chance that you could check with your science teacher and see if they have any at the school lab. (I will get back to you on the calculation in a separate post.)

3 & 4 - Let's do the easy part first - Yes, the part you have picked out will allow you to insert tube A in tube B. (You want to check for that the Outer Diameter of Tube A is equal to or smaller than the Inner Diameter (ID) of tube B). Now, onto the more difficult part, we need to find out whether PVC is compatible with the chemical that you plan to use in this experiment. (Please try the search term on Google "PVC chemical compatibility" and then you will look for the chemical such as "Ferric Nitrate" to make sure that it is compatible. If it is not, we may need to use a different type of tubing material such as Tygon.)

Ok, I will continue to work on the list of questions and continue to post as I finish them.
Good Luck!
Willey

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Re: Help Needed: Experiment with Coccolithophorid Algae

Postby SciB » Wed Aug 07, 2019 6:05 pm

Hi Lakshita,

I would like to offer my help with your algae project as Mr Willley, whose answers so far have been perfectly fine, wished to have a biological expert (which I am) join the forum.

I have read all your posts and Willey’s answers and I understand that you are asking the question—what is the best iron (Fe3+) concentration for coccolithophore algae growth. So, the independent variable will be the ferric nitrate concentration and the dependent variable will be the amount of CO2 taken up by the algae over a certain period of time.

From reading your posts, I assume that you have a source of the algae (Emiliana huxleyi ?) from Ward’s Scientific Supply and can also have your school order the ferric nitrate, or use theirs if they have it. You are working on building the culture bottles for the algae and that seems to be going OK, but I’m sure you will have more questions.

You did have a question about the concentration of the ferric nitrate and I can help you with that. One part per million, ppm, of any chemical in water solution is equivalent to one milligram (mg) per liter (L). One milliliter (mL) of water has a weight of 1 gram (1000 mg), so 1L of water is equivalent to 1 million mg, and 1 ppm = 1 mg/L.

The recipe in the procedure section of the online coccolithophore experiment (http://www.odec.ca/projects/2008/sidd8g2/index.html) tells you to dissolve 10.3 g (10,300 mg) of ferric nitrate in 250 mL of distilled water. Dissolving that much ferric nitrate in 250 mL of distilled water gives you a solution that contains 41,200 mg/L or 41,200 ppm, which would be much, much higher than the concentrations given in the CO2 fixation project: 0.024 to 0.38 ppm (24 to 380 ppb). There was no explanation of why these iron concentrations were chosen. There is a bibliography of links but they are not connected to anything in the text as they are in a scientific paper and most of them are useless as they no longer work.

Here’s a link to an article about iron concentrations in seawater: https://www.google.com/url?sa=t&rct=j&q ... 1mkJ36ljdn

I did a search for the average concentration of iron in seawater and found it to be 10-100 ppb (parts per billion), but this really does not tell you an optimal iron concentration for growth of E. huxleyi. If the iron concentrations in the online experiment are converted to ppb they are 24 to 380 ppb, which sounds reasonable. However, the amount of ferric nitrate added to the water in the procedure is off by about 1 million according to my calculations. A 1 ppb ferric nitrate concentration means 1 microgram (ug) per liter. Micrograms are too small to weigh out, so scientists routinely make a more concentrated solution and then dilute it to get the correct final concentration. So, if you dissolved 24 mg of ferric nitrate in 1 L you would have a 24 ppm solution. Then, if you put 1 mL of that solution into 1 L of algae growth medium, the final concentration would be 24 ppb—the correct amount. For the higher concentrations, just add a larger volume—2 mL for 48 ppb, 4mL for 96 ppb, 8 mL for 192 ppb and 16 mL for 384 ppb.

I hope this does not confuse you. Try the calculations yourself and ask your teacher to verify them and you will hopefully find that they are correct.

I’m sure you will have more questions, so I will be waiting for your next post.

Good luck!
Sybee

LeungWilley
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Re: Help Needed: Experiment with Coccolithophorid Algae

Postby LeungWilley » Thu Aug 08, 2019 5:55 pm

Hi Lakshita,
Continuing on the list:
5. Air Pump, In-line air filter, hose fitting, multiport manifold.
I am sorry to say that I am not seeing much from Home Depot that would be suitable for use in this experiment. In fact, I would lean more towards a fish tank air setup. (You might want to pay a visit to a pet store and see if they can help you with a setup)

At any rate, here's my attempt at creating this setup:
https://smile.amazon.com/Mylivell-Quietest-Aquarium-Silent-Silicone/dp/B01MTSR8Y8/ref=sr_1_4?keywords=inline+air+filter+fish+tank&qid=1565229170&s=gateway&sr=8-4

The draw back with this pump is that you are going to need to an adapter to go from 1/4" tubing to the 3/16" port on the pump. You can use something like this:
https://www.amazon.com/Metalwork-Reducing-Splicer-Fitting-Reducer/dp/B07C2ZCT97/ref=sr_1_5?crid=2THSAQ30K5CFF&keywords=3%2F16+to+1%2F4+hose+barb&qid=1565311434&s=industrial&sprefix=1%2F4+-3%2F16+hose%2Cindustrial%2C152&sr=1-5

For the Air Manifold, you might want to look at something like this:
https://www.amazon.com/Interstate-Pneumatics-FPM44S-Aluminum-Rectangle/dp/B002RRRIOK/ref=sr_1_14?keywords=air+manifold&qid=1565310962&s=gateway&sr=8-14

And then I would suggest pairing this manifold with these "push to connect" fittings.
https://www.amazon.com/dp/B07LBJTVP5/ref=nav_timeline_asin?_encoding=UTF8&psc=1

If you were to use something like this, you can eliminate the nuts since the tubing will be secured via the push to connect fittings.

6. See answer to 5

7. Yes, you should be able to get / borrow a microscope from the schools around your area. Here's one from Wash U:
https://schoolpartnership.wustl.edu/items/wolf-cordless-inclined-elementary-microscope/

Also, if you prefer, you may also want to look at a USB microscope that connects to your phone or school laptop / chromebook, etc... (It might make it easier for your eventual report / presentation)
https://www.amazon.com/Jiusion-Magnification-Endoscope-Microscope-Compatible/dp/B06WD843ZM/ref=sr_1_3?keywords=usb+microscope&qid=1565311655&s=gateway&sr=8-3

Ok, I think that sums up what I was able to find.
I hope this helps and best of luck with your experiment!

Willey

P.S. Thanks Sybee for jumping in to help with this topic!

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Re: Help Needed: Experiment with Coccolithophorid Algae

Postby SciB » Thu Aug 08, 2019 7:14 pm

Hi Willey,
I am happy to help get the science right and the experiments working correctly.

The corrected amounts for the addition of ferric nitrate solution to the algae cultures needs to be checked and verified but I believe that my calculations are correct. The ocean levels of iron are in the ppb range and the amount of ferric nitrate given in the online experiment procedure would have been way higher than what was stated.

Hi Lakshita,
Please let me know if you have questions about my ferric nitrate calculations. Is there a chemistry teacher at your school? If so, I would recommend that you call them and ask if you can come and have them check the calculation. A chemistry teacher would be the person most familiar with this type of calculation and could verify the amounts and volumes.

I will be happy to assist you with any other parts of your project. There are a lot of important details to pay attention to in this set of experiments and it would be great if you would write up the procedure ahead of time and post it so I can check it and let you know if anything needs to be changed. One error in the procedure can ruin an entire science project.

If possible, you should do three cultures for each ferric nitrate concentration so you will be able to average the CO2 readings and do a statistical comparison on the results. This is the only way that you can prove your conclusions. If you need help with statistics there are a lot of good tutorials on Youtube and I will answer any of your questions.

If you feel like this would be too many cultures to run at one time, you could eliminate a couple of the ferric nitrate concentrations. You could do a zero concentration control of just seawater and then three Ferric nitrate concentrations--low, medium and high. It is statistically better to do replicates of one concentration than single readings of more concentrations. No scientific research is accepted without statistical analysis and confirmation and you should follow that rule.

Hope to hear from you soon,

Sybee

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Re: Help Needed: Experiment with Coccolithophorid Algae

Postby lakshu_s » Fri Aug 16, 2019 12:30 pm

Hi Mr.Willey/Sybee,

Thanks so much for your clarifications. Sorry I missed all the posts since , I started my high school this week and was busy getting accustomed to new atmosphere/schedule . I will go over all the clarifications during the weekend and post my questions. With your guidance, I am so confident that I can finish my project on time.

Thanks,
Lakshita


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