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Re:Culture Journal, Species: Tetraselmis
Thursday, March 22, 2012 12:09 PM
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This got me thinking a bit further, and asking myself the question, "How does the amount of nitrogen I am putting into the tank with a liter of phytoplankton compare to the amount of nitrogen I am putting into the tank with a cube of frozen food?"
The nitrate in 1 liter of F/2 media, expressed as absolute milligrams (and not a concentration), is appx. 55 milligrams.
The typical cube of frozen food is approximately 3 grams in wet weight. Typical frozen food contains between 5 and 15 percent of the wet weight of protein, so for the purposes of this exercise, let's call it 10%. Protein contains, on average, 16% nitrogen. That means that the typical cube of frozen food contains 3 g * 10% * 16% = 48 mg of nitrogen. The molecular weight of the nitrate ion is 62.44, and the atomic weight of nitrogen is 14, so that means that the 48 mg of nitrogen would convert into 48 * 62.44 / 14 = 214 mg of nitrate.
1 typical cube of frozen = 214 mg of nitrate.
1 liter of F/2 = 55 mg of nitrate.
So, adding 1 cube of typical frozen food adds roughly 4 times the potential amount of nitrate as 1 liter of F/2 media. This is assuming, of course, that all of the nitrogen in each ends up being ultimately converted into nitrate.
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Re:Culture Journal, Species: Tetraselmis
Friday, March 23, 2012 7:41 AM
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Weird, I replied to this and even saw my post! Now it is gone!
I said thanks for posting the math, Jim! That makes a lot of sense. I understand that nitrogen going into the tank in any form is still nitrogen. It's the same as when nuisance algae is growing and it dies off. You're better off removing it than killing it. The math regarding the fertilizer used for phyto is so miniscule though that it really doesn't seem to have any threat at all.
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Re:Culture Journal, Species: Tetraselmis
Sunday, March 25, 2012 7:55 AM
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I think last time I bought Sodium thiosulphate I bought it at a drug store, so I'm going to see if I can find some there today as well as some ethanol to make a filter. I think I better split it today, it sure is dark! Like soup... 
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Re:Culture Journal, Species: Tetraselmis
Sunday, March 25, 2012 9:51 AM
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Definitely time to split. Got that microscope fired up yet?
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Re:Culture Journal, Species: Tetraselmis
Sunday, March 25, 2012 11:01 AM
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I'm working on firing up the microscope this morning. Right now I'm working on a sterile split, and wondering...how long do I microwave the medium for? I can't find reference.  EDIT: I microwaved 2 liters of medium for 8 minutes which made it quite warm, but not hot. I'm not sure if it is supposed to boil or maybe just the microwaves have a sterilizing effect...? I just took a video and am uploading to You Tube. Unfortunately the best I can do is 185x magnification which doesn't look like enough to ID it. What magnification do you guys use for phyto? 1000x? EDIT 2: Ok, so upon searching I find that ethanol is what is in isopropyl rubbing alcohol. I have 70% isopropyl rubbing alcohol. Is this sufficient to make a cotton filter with? Do I just use "cotton balls"?
<message edited by EasterEggs on Sunday, March 25, 2012 11:59 AM>
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Re:Culture Journal, Species: Tetraselmis
Sunday, March 25, 2012 12:52 PM
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I microwave until the media is 185F / 85C. I find that the media does boil in the microwave, even though it is not up to "boiling temperature". I microwave 2 x 2-liter bottles that each contain 1500 ml of media for 22:22 in an 1100 watt microwave, and that does the trick for me.
Isopropyl rubbing alcohol contains isopropyl alcohol, not ethanol. It is probably good enough to kill off the things in the cotton, but the 30% water will also probably take longer to evaporate. I used 100% denatured ethanol from the drug store. Denatured ethanol contains ethanol and a little bit of nasty-tasting things that make it so nobody wants to drink it.
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Re:Culture Journal, Species: Tetraselmis
Sunday, March 25, 2012 12:56 PM
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Ok, thanks for clarifying the microwaving strategy and the ethanol. Google must be wrong about isopropyl alcohol being ethanol. Ha! Here is my first split, done as sterile as I know how. No air filter yet though. 
<message edited by EasterEggs on Sunday, March 25, 2012 1:08 PM>
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Re:Culture Journal, Species: Tetraselmis
Sunday, March 25, 2012 5:40 PM
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So I went to the pharmacy and asked for the purest ethanol they have. I was given a bottle that says "Rubbing Alcohol Compound". Medicinal ingredient: 95% Ethanol Anhydrous. Mfr. Standard. Non-medicinal ingredients (alphabetical order): Camphor, Diethyl Benzyl Benzoate, and Diethyl Phthalate. It was like $3. Is this the right stuff?
I had absolutely no luck finding sodium thiosulphate...maybe I should have included it in my FAF order! I figured I could buy it locally...
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Re:Culture Journal, Species: Tetraselmis
Sunday, March 25, 2012 7:33 PM
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That's funny. I thought Rubbing Alcohol was Isopropanol, not Ethanol. If you want ethanol, use Vodka.
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Re:Culture Journal, Species: Tetraselmis
Sunday, March 25, 2012 7:40 PM
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 Originally Posted by EasterEggs
..l. Google must be wrong about isopropyl alcohol being ethanol. Ha!
...
Ethanol has 2 carbon atoms, Propanol has 3, and isopropanol or isopropyl alcohol has 3 carbons in a V configuration, with the OH alcohol part attached to the carbon in the middle. I majored in Chemistry, but this is high school stuff. I'm surprised at Google…. http://en.wikipedia.org/wiki/Isopropyl_alcohol
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Re:Culture Journal, Species: Tetraselmis
Sunday, March 25, 2012 8:22 PM
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The "Rubbing Alcohol Compound" doesn't mention isopropyl at all, and lists "ethanol" as 95%. Funny you mention it Kathy, I have been reading some vodka dosing articles lately and have seen it referred to as "ethanol dosing" on several occasions. I'm just wondering if this 95% ethanol "Rubbing Alcohol Compound" will be sufficient for an air filter for my phyto? Or maybe soaking the cotton in vodka is good enough? Haha! Wikipedia says, " Rubbing alcohol, USP / B.P. is a liquid prepared and used primarily for topical application. It is prepared from a special denatured alcohol solution and contains approximately 70 percent by volume of pure, concentrated ethanol (ethyl alcohol) or isopropyl alcohol (isopropanol). [1] Individual manufacturers can use their own "formulation standards" in which the ethanol content usually ranges from 70-99% v/v. [2] In Ireland and the UK, the equivalent skin preparation is surgical spirit, which is always an ethyl alcohol-isopropyl alcohol mixture." http://en.wikipedia.org/wiki/Rubbing_alcohol Which makes me think that "rubbing alcohol" could be isopropyl or ethanol. Wiki also says, " Denatured alcohol or methylated spirits is ethanol that has additives to make it more poisonous or unpalatable and, thus, undrinkable. In some cases it is also dyed." http://en.wikipedia.org/wiki/Denatured_alcohol
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Re:Culture Journal, Species: Tetraselmis
Sunday, March 25, 2012 11:39 PM
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Originally Posted by EasterEggs The "Rubbing Alcohol Compound" doesn't mention isopropyl at all, and lists "ethanol" as 95%. Funny you mention it Kathy, I have been reading some vodka dosing articles lately and have seen it referred to as "ethanol dosing" on several occasions. I'm just wondering if this 95% ethanol "Rubbing Alcohol Compound" will be sufficient for an air filter for my phyto? Or maybe soaking the cotton in vodka is good enough? Haha! Wikipedia says, " Rubbing alcohol, USP / B.P. is a liquid prepared and used primarily for topical application. It is prepared from a special denatured alcohol solution and contains approximately 70 percent by volume of pure, concentrated ethanol (ethyl alcohol) or isopropyl alcohol (isopropanol). [1] Individual manufacturers can use their own "formulation standards" in which the ethanol content usually ranges from 70-99% v/v. [2] In Ireland and the UK, the equivalent skin preparation is surgical spirit, which is always an ethyl alcohol-isopropyl alcohol mixture." http://en.wikipedia.org/wiki/Rubbing_alcohol Which makes me think that "rubbing alcohol" could be isopropyl or ethanol. Wiki also says, " Denatured alcohol or methylated spirits is ethanol that has additives to make it more poisonous or unpalatable and, thus, undrinkable. In some cases it is also dyed." http://en.wikipedia.org/wiki/Denatured_alcohol I think Rubbing alcohol COULD be Ethanol, I've just never encountered it as such. I would be more comfortable using something that is less toxic generally than whatever they are putting in rubbing alcohol to make it unpalatable. (I do know of an alcoholic who drank Listerine until it nearly killed him, in an effort to remain inebriated without smelling of alcohol, and alerting his long suffering wife to his fall from sobriety. ) I only wanted to make it clear that isopropanol is not ethanol. For the purposes of sterilizing cotton for an air filter, I would use ethanol, vodka is probably cheapest. Not sure what those other things in rubbing alcohol would do to the phyto.
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Re:Culture Journal, Species: Tetraselmis
Monday, March 26, 2012 10:26 AM
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I think a concern here is not only using something that will sterilize the cotton, but also something that will evaporate away quickly. My concern about using Vodka (40% ethanol and 60% water) is that the water will persist for some time, perhaps encouraging the growth of bacteria, fungi, etc., in the cotton. Using 100% alcohol (denatured or not), whether ethanol or isopropyl (or even methanol, for that matter!) will greatly decrease the chance of lingering water in the cotton allowing the very same microbes we are trying to filter out to flourish.
Between vodka and 70% isopropyl, I'd go with the 70% isopropyl. For me, it was very easy to go to the nearest drug store and get 100% denatured ethanol.
EDIT: You know, in practical terms, you could probably dispense with the ethanol altogether, and just run air through the filter for 24-48 hours to "flush" out any microbes hiding in the cotton. If you pack it tight enough, this is probably a non-issue, and the ethanol is just an extra, but not absolutely necessary, precaution. I would fear a soggy wet filter from residual water more than I would one that was dry, and had had no alcohol treatment at all.
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Re:Culture Journal, Species: Tetraselmis
Monday, March 26, 2012 7:17 PM
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Thanks for the input Kathy, I was also worried about the other ingredients in the 95% ethanol.
Jim, what about the 95% ethanol rubbing alcohol? Would you take the 70% isopropyl over the 95% ethanol?
You bring up a good point about soggy cotton Jim, I could easily do an experiment packing a filter, soaking in vodka or isopropyl, or the ethanol, let it run and then take it apart in an hour or so and see if it is dry or not.
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Re:Culture Journal, Species: Tetraselmis
Wednesday, March 28, 2012 8:25 AM
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I need a bigger air pump on the pop bottle. I guess it is time to hook up the Coralife Super Luft SL-65 pump in the broodstock room. That way I can steal the bigger Hagen pump for the phyto station.
I'm going to do what I always like to do...I'm going to crash one of the cultures. I will split the cup culture tonight (it's dark again already), and will leave it in front of the lights until it crashes. I like to test cultures that are new to me so I can learn where the crash point is and if I am able to bring a culture back from a partial crash. I think these are good things to know.
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Re:Culture Journal, Species: Tetraselmis
Tuesday, April 3, 2012 9:41 AM
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So pop bottle #1 was start on March 25th so it is 9 days old today and has been very dark (can't see through it) for several days now. This morning it suddenly settled out real bad so that I could just barely kind of see through it. I assume this is the start of a crash?
I poured a third of the bottle down the drain (no empty pop bottles), and topped back up with sterile medium. I will get more pop bottles today so I don't have to waste the phyto when I'm trying to ramp up! That was annoying dumping it down the drain although now I think about it I probably could have saved it in a bucket for the day. Argh.
In the future, when a phyto culture settles out like that (assuming this is the start of a crash), should I only split the phyto that is still motile? Should I toss the bottom of the bottle that settled?
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Re:Culture Journal, Species: Tetraselmis
Sunday, May 27, 2012 8:07 AM
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These cultures are still just plugging along. I find sometimes after I split the new culture fails to darken and falls out of the water column in a week or so. I just chuck it out and make a new one. I need to purchase some more lights and get another shelf going. I currently have (2) 24" 13W T5 on the one shelf, and I'm wondering if (1) T5 on each shelf would be enough?
<message edited by EasterEggs on Sunday, May 27, 2012 10:09 AM>
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Re:Culture Journal, Species: Tetraselmis
Friday, June 1, 2012 6:30 AM
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The Tetraselmis has been having a tough go lately (not darkening, kind of yellowing a bit) since the temps outside have warmed up into the mid 20s (Celcius). I checked temps in the phyto closet at the warmest part of the day (lights were off) and noted 86F. Ok, that's a tad warm! I had the lighting cycle reversed to daylight thinking that would keep temps down in there, but it didn't. So now I switched back to daylight light cycle and I'm opening the door of the closet while the lights are on and that is keeping temps closer to 81-83F. I also moved the phyto bottles 6" away from the lights instead of the 2" they were. The phyto is finally starting to darken again.
<message edited by EasterEggs on Sunday, July 1, 2012 9:11 AM>
Don't let fear and common sense stop you! =]
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Re:Culture Journal, Species: Tetraselmis
Sunday, June 17, 2012 6:42 PM
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That darkening didn't last long, the Tet started to lighten/yellow a tad again. I haven't had to split in over a month, so it must be barely hanging on. Today, I split the cultures to freshen them up and put them through a 23 micron sieve. I got some yellowish "silt" (critters or deposits I'm not sure) in the sieve. Hopefully these cultures revive soon!
Don't let fear and common sense stop you! =]
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Re:Culture Journal, Species: Tetraselmis
Sunday, July 1, 2012 7:10 AM
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There was still no revival of the Tetraselmis, and I haven't had a culture darken in at least 6-10 weeks. So I added a fan to the phyto shelf. I've also been trying to keep the fish room a few degrees cooler. With these changes I have been able to keep the bottles of Tet just under 80F. I added the fan to the right hand side of the shelf, and you can see in the photo that the Tet is (finally!) darkening with the right culture the best looking, and the left culture still very light. The one on the left is how these cultures have looked for the last 6-10 weeks! I now moved the fan to the middle of the shelf where it can blow directly on the cultures. I'm hoping I have finally found the solution! 
Don't let fear and common sense stop you! =]
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