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Culture Journal, Species: [isochrysis galbana]
Sunday, May 24, 2015 4:34 PM
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Culturing Journal DataSheet This first post should be updated regularly to include new information as events take place or changes are made to your system General Species: isochrysis galbana Species description: motile brown algae Culture source (link if possible ): If algae, CCMP # (Optional ): http://ccmp.bigelow.edu/ Culture Establishment Date: July 30, 2014 Continuation Date: still going 7/3/2018 http://i893.photobucket.com/albums/ac132/aquatictec_photos/MBI/isoculture20150524-2.jpg Culturing Vessel Details Salinity: 30.5ppt Temperature: 72-78 pH: never measured Vessel description: 6 gallon clear plastic carboys Lighting description: 72 watt led light bar 2 per 4' or 4 bottles Lighting cycle: 24/7 continuous Aeration description: filtered outside air vigorous aeration with rigid airline tubing and central blower Methodologies Split methodology: once per week 50 percent since we started using CO2 we keep 1 gallon to start a new one and use the other 5 gallons weekly since CO2 we keep 1 gallon and harvest 5 weekly per culture Culture medium description: sterilized salt water with 2.5 ml per gallon of Guillards F/2 formulation per gallon currently 9ml per 6 gallon Cell count: not performed dark brown color when split Reference links: Additional Information (No Pictures or Videos in the Section Please) Notes: we use fresh saltwater made with red sea salt. We keep 7 cultures going. We sometimes loose some of them but we just split the remaining ones You will be required to provide photographic evidence and as much detail as possible about your project in this thread. If your thread does not contain detailed enough photos and information the MBI Council will not be able to approve your reports.
<message edited by aquatictec on Tuesday, July 03, 2018 7:27 PM>
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Re:Culture Journal, Species: [isochrysis galbana]
Tuesday, July 05, 2016 7:29 PM
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Re:Culture Journal, Species: [isochrysis galbana]
Tuesday, July 05, 2016 7:32 PM
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above picture was taken today July 5th, 2016 We are using 2 led light bars one is white and the other red/blue The red causes a green cast on the photos The culture has been going for almost 2 years We do loose some every now and then but we keep more cultures than we need going
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Re:Culture Journal, Species: [isochrysis galbana]
Tuesday, July 05, 2016 8:07 PM
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Why are you culturing it? I mean, what microfauna are you culturing as well?
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Re:Culture Journal, Species: [isochrysis galbana]
Monday, July 11, 2016 3:17 PM
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Re:Culture Journal, Species: [isochrysis galbana]
Wednesday, July 27, 2016 6:14 PM
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We have sometimes lost cultures when we were splitting them. We are using 2 72 watt LED lights per 4 or 5 six gallon carboys The phyto was probably getting killed from the much higher light intensity We just started turning one light bar off when we split the culture and turning it back on in a day or 2
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Re:Culture Journal, Species: [isochrysis galbana]
Wednesday, July 27, 2016 7:22 PM
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Do you pasteurize your medium and if so, how?
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Re:Culture Journal, Species: [isochrysis galbana]
Wednesday, July 27, 2016 8:23 PM
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the water for culture is treated with bleach and dechlor the water for the fertilizer is boiled then cooled. We keep a bag over the bottle and keep it in the refrigerator It lasts 3 or 4months so there may be a problem there but we occasionally put some fertilizer in sterile water to see if anything grows and nothing has
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Re:Culture Journal, Species: [isochrysis galbana]
Sunday, September 04, 2016 9:11 PM
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can you elaborate on how much chlorine you add to sterilize your water? curious how to sterilize water with bleach
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Re:Culture Journal, Species: [isochrysis galbana]
Monday, September 05, 2016 7:20 PM
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8ml 5.25 percent bleach per 5 gallons as of 7/3/2018 we use 9ml
<message edited by aquatictec on Tuesday, July 03, 2018 7:28 PM>
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Re:Culture Journal, Species: [isochrysis galbana]
Saturday, November 26, 2016 4:43 PM
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11/26/2016
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Re:Culture Journal, Species: [isochrysis galbana]
Thursday, January 04, 2018 11:06 AM
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About a month or so ago we started having problems with our cultures. They were contaminated with what appeared to be a ciliate about 20 microns by 5 microns with "hairs" about 5 microns long. Because of the contamination they were not growing very rapidly. We ordered a new dense 2 liter culture from algagen and also strained our existing cultures through a 53 ish micron mesh. (I am not sure of the exact size) Usually the cultures from algagen take off but it just kept very slowly growing as did our old cultures. I split the culture from algagen with another institution and it did not take off for them either. I do not know where the contamination came from and how it hit all of our cultures. I am pretty sure it was because someone did not sterilize their hands. We used to shake the cultures. We have since quit that. I have not let any one else touch the cultures since then. I have also placed lexan shields between the cultures Obviously the mesh size was not sufficient to get all the ciliates. Kathy Leahy referred me to an article that said quinine sulfate could be used to kill ciliates in phyto cultures. We tried it at a dose of 10mg/liter in our iso cultures and also in a nano culture. At that dose it killed all the ciliates. It also killed the iso but did not kill the nano. Just recently our old contaminated cultures and the culture from algagen started growing rapidly. I have some 10 micron screening on order. I will try using that on our contaminated cultures. We are also growing out the culture from algagen.
<message edited by aquatictec on Tuesday, November 13, 2018 8:14 PM>
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Re:Culture Journal, Species: [isochrysis galbana]
Friday, January 19, 2018 6:43 PM
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I kept one culture that had been dosed with quinine sulfate. It appears that it is coming back. It will be interesting to try dosing this culture with a small sample of quinine sulfate and see if it fares as poorly as last time. (The engineer in me keeps popping up) The cultures from algagen finally took off. It must have experienced some shock in shipping.
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Re:Culture Journal, Species: [isochrysis galbana]
Tuesday, March 13, 2018 6:08 PM
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We purchased a new culture from algagen. It never did well. We have been babying it. I split it with another institution and it never did well for them either. I am trying purifying it by streaking it on a .5 percent agar plate. I have some small flasks on order. The plan is to try growing some of the plated iso out first in a test tube shaken daily and then when/if it browns up to 50 ml flask with aeration then 500 ml etc. We will also dilute some of the plated agar and start a new plate. We also have some more iso on order. We have to do something to make sure we always have iso available. I want to remove its availability from the equation. If the agar plate thing works out we will probably just start purify a new culture every so many weeks and grow them out.
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Re:Culture Journal, Species: [isochrysis galbana]
Tuesday, March 13, 2018 6:33 PM
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double post not sure how to delete
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Re:Culture Journal, Species: [isochrysis galbana]
Tuesday, June 26, 2018 3:18 PM
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agar plating works quite well on iso We have been able to start new cultures by streaking an existing culture on an agar plate and starting new cultures from cells isolated from the plate. In early stages we get doubling in less than a day
<message edited by aquatictec on Tuesday, July 03, 2018 7:29 PM>
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Re:Culture Journal, Species: [isochrysis galbana]
Wednesday, September 19, 2018 5:48 PM
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We had a culture of iso that we had scaled up from a cell to 200 ml about 2 months ago. We just left it in a 2l flask with 2 rigid airlines and a cotton filter on top. It has stayed nice and golden. Yesterday I boiled 1800 ml of salt water let it cool then added fertilizer. I added 200ml of this solution to the ancient iso culture. I didn't want to shock the culture with a huge s.g. difference. Today the culture looked nice and golden so I added the rest of the fertilized water. We will see what happens.
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Re:Culture Journal, Species: [isochrysis galbana]
Tuesday, November 13, 2018 8:12 PM
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We had no problem scaling it up. Since then we have started test tubes from plates that had been sitting under light for over 3 months. They also scaled up nicely. Iso is getting awful easy to cultivate !
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