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Culture Journal, Species: Oxyrrhis marina
Thursday, August 11, 2011 7:14 AM
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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: Oxyrrhis Marina Species description: dinoflagellate Culture source (link if possible ): Jim Welsh, kind donation. If algae, CCMP # (Optional ):CCAP# 1133/3 Oxyrrhis marina Dujardin http://www.algaebase.org/...tail/?species_id=52504 http://ccmp.bigelow.edu/ Culture Establishment Date: 6 /8/2011 Continuation Date: Culturing Vessel Details Salinity: 20ppt Temperature: ambient pH: 8ish Vessel description: plastic jars of various sizes, gallon and half gallon, and 5 gallon buckets Lighting description: fluorescent grow bulb on timer Lighting cycle: 14 day , 10 night Aeration description: open airline, few bubbles per second, rapid but not boiling Methodologies Split methodology: various. I've previously kept them going with just feeding and weekly additions of saltwater. And then they crashed. Now wish to try more dynamic method. Daily removal of part of the culture, about 1/8 initially, replacement with 2 x the volume removed of clean saltwater, 20ppt. Feeding twice daily. Culture medium description: Bleached saltwater is dechlorinated with excess chloramX and RotiGrow + and or N-Rich added as need to maintain or grow culture. Successful method follows: Cell count: I am not able to count them. Qualitative observations ? I can see them swimming in the 40x objective of the scope. Reference links: http://www.int-res.com/ar.cles/ame/10/a010p307.pdf Grazing in the heterotrophic dinoflagellate Oxyrrhis marina: size selectivity and preference for calcified Emiliania huxleyi cells http://www.algaebase.org/...tail/?species_id=52504 Additional Information (No Pictures or Videos in the Section Please) Notes: 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 KathyL on Tuesday, August 23, 2011 9:48 PM>
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Re:Culture Journal, Species: Oxyrrhis marina
Thursday, August 18, 2011 5:53 PM
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Well, I had a thriving culture in a 1 gallon jar. I mostly just fed it with Andy's mix of 6 parts N-rich + 1 part RotiGrow+, and an equal volume of ChloramX. Not a lot, though, perhaps less than 1 ml per day, divided over 2 times. I periodically added a half cup or so to the other cultures I am keeping: M. salina and A. panamensis. This morning, I took a sample to look under the scope. My "pure" culture of O. marina was completely dead. My culture of A. panemensis had a nice co- culture of O. marina, so I used a 37 micron screen to capture the Panamensis and return to its own culture, and let the O. marina through, into a clean jar, fed, aired, and off we go again. This time I'll keep better records of how much I feed, water changes, etc.
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Re:Culture Journal, Species: Oxyrrhis marina
Thursday, August 18, 2011 10:19 PM
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Correction: My culture of A. panemensis had a nice co- culture of CILIATES, so I used a 37 micron screen to capture the Panamensis and return to its own culture, and let the CILIATES through, into a clean jar, fed, aired, and off we go again. Tomorrow I'll scrap it and get the O. marina from the Enrichment bucket.
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Re:Culture Journal, Species: Oxyrrhis marina
Saturday, August 20, 2011 6:24 PM
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I decided to keep the ciliates in their own culture. They are second from the right in the picture in the next post. I started what I hope is O. marina from the tinier swimming dots in my enrichment rotifer bucket. They seem to be doing well. They are in a 1 gallon jar with air, only filled about 1/3 of the way with cuture at the moment. They are presently being fed with 6 drops of "6NR/1RG+" (6parts N-Rich, 1 par RotiGrow+), twice per day, and an = volume chloramX. Next week's PLAN: Remove one pudding cup of O marina to M. salina and A. panamensis cultures each. Feed with 3 cups of clean saltwater, and 6 drops of 6NR?1RG+ twice daily. This should slowly increase volume. At end of week decant culture to new clean gallon jar.
<message edited by KathyL on Tuesday, August 23, 2011 6:20 PM>
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Re:Culture Journal, Species: Oxyrrhis marina
Tuesday, August 23, 2011 6:18 PM
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Here's the culture shelf. O marina on the right.
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Re:Culture Journal, Species: Oxyrrhis marina
Tuesday, August 23, 2011 9:30 PM
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ChloramX recipe: 40 grams of dry chloramX in 500 ml clean water. This is kept in the refrigerator until use. I use it in cultures by adding an equal volume of it to the volume of algae food I add. I also use it to dechlorinate the bleached water change water I use for culture medium. Here's what I am doing: Discarded water from broodstock water changes is stored in a clear plastic 17 gallon storage tub, diluted to 20 ppt, pH checked and chlorinated with 1 ml bleach per gallon. The tub is fixed with a bulkhead and a plastic faucet, and is covered with a plastic lid that does not seal. I can draw off a gallon at a time of the bleached water, dechlorinate with a bit more than 1 ml of my refrigerated chloramX stock, and test for chlorine, and use it to dilute and feed my cultures and also rotifer culture.
<message edited by KathyL on Tuesday, August 23, 2011 9:51 PM>
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Re:Culture Journal, Species: Oxyrrhis marina
Wednesday, August 24, 2011 12:32 PM
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FWIW, Here's a more detailed explanation of proper ClorAm-X dosing that Eric Henry of Reed Mariculture emailed me:
Originally Posted by Eric Henry via Email
Hi Jim- ClorAm-X is wonderful stuff, in fact it is the key to high-density rotifer cultures, but it is possible to misuse it. The preferred method of use is to measure ammonia, and dose accordingly. Contrary to the statements of the manufacturer (and not surprisingly), we have found that it is in fact possible to overdose and inhibit the growth of rotifers, and no doubt other animals. This has happened when users have blindly thrown in ClorAm-X in reaction to finding a high level of ammonia in their cultures. For rotifers inhibition becomes evident at about 0.3 g of unreacted ClorAm-X per liter of culture. Of course you can put in more ClorAm-X than this, as long as there is enough ammonia to react with it and “use it up”. But nobody wants to have to measure ammonia every day. We have found that our continuous rotifer cultures are stable and predictable, and we have learned that for a given feed dosing we need a certain dosing of ClorAm-X. For other culture systems the dosing is likely to be different. But here is a rule-of-thumb to start with: Rotifers, at least, can be expected to convert about half of the nitrogen in the feed to ammonia (the rest goes into rotifer proteins). Our algae pastes are all about 50% protein, on a dry weight basis. The dry weight of the paste is roughly half of the first 2 digits of the number in the Instant Algae product names, so Nanno 3600 is 18% dry weight. It works out like this: Nanno 3600 is 180 gm per liter dry weight X 52% protein X 16% nitrogen in protein = 15 gm/l nitrogen = 18.2 gm ammonia /l = 18.2 mg ammonia /ml. But if only half the nitrogen goes to ammonia, then 9.1 mg ammonia/ml of Nanno 3600 will be produced. Dosing formula is 31.9 mg ClorAm-X to neutralize 1 mg ammonia, so you would need to add 290 mg ClorAm-X per ml of Nanno 3600 that you feed, or 290 gm/liter. But this is a “worst-case scenario”. Could the protein conversion rate be better than 50%? Will your animals eat all the algae? Are you removing a lot of solid wastes from the culture (this removes some nitrogen)? If you do a 25% water change, it removes 25% of the ammonia. The natural bacteria in your cultures will convert some of the ammonia to harmless nitrate. So to calculate your dosing, first consider the dry weight of the feed you are using. For our non-Instant Algae feeds: Rotifer Diet is about 5% dry weight, PhytoFeast is about 2.8% (we thin these out because the Instant Algae pastes are too thick to be accurately dosed in the very small amounts of biomass that reefers should add to their tanks in one feeding). Your suggested dosing of 12 teaspoons for 1 liter is equivalent to 60 g/L (1 tsp = 5 gm). PhytoFeast would take 45 g/L Rotifer Diet would take 80 g/L In theory! Really the practical safe ClorAm-X dosing can only be prescribed in relation to the ammonia load, which really must be determined by testing. This is especially true if you dose ClorAm-X day after day and only do partial water changes—this can allow an excess to gradually accumulate to inhibitory levels. Of course nobody wants to waste money either, also a good reason for not using more than you need. We have seen no evidence that ClorAm-X solutions break down chemically, but the less concentrated they are, the more likely that fungi can grow in them. Refrigeration is a good idea in any case. I suggest that you not make up more than you can use in a couple of weeks. . Eric C Henry, Ph.D. Reed Mariculture Inc.
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Re:Culture Journal, Species: Oxyrrhis marina
Wednesday, August 24, 2011 6:11 PM
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One other thing to note: API ammonia test kits are the best one we have found that will actually work with ChlorAm-X.
<message edited by GreshamH on Tuesday, November 1, 2011 7:06 PM>
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Re:Culture Journal, Species: Oxyrrhis marina
Wednesday, August 24, 2011 10:25 PM
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Thanks Gresh!
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Re:Culture Journal, Species: Oxyrrhis marina
Thursday, August 25, 2011 10:07 AM
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Originally Posted by
I mostly just fed it with Andy's mix of 6 parts N-rich + 1 part RotiGrow+, and an equal volume of ChloramX. Not a lot, though, perhaps less than 1 ml per day, divided over 2 times. I'm worried that you aren't feeding enough, Kathy. I feed 6 parts N-rich + 1 part Rotigrow _twice_ per day. Total daily is 12 parts N-rich + 2 parts Rotigrow.
--Andy, the bucket man. "Not to know the mandolin is to argue oneself unknown...." --Clara Lanza, 1886
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Re:Culture Journal, Species: Oxyrrhis marina
Thursday, August 25, 2011 1:08 PM
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You are surely right. It does depend on the density of the culture, and mine is probably not as dense as yours. I am slowly increaseing the food volume as I see the culture clearing. I am now dosing at twice the rate I dosed when I wrote this. When I write about 6 parts and one part, I am talking about the ratio of one food to the other. I mix it up in a 3 oz bottle and use that for about a week. When I say I dose 1 ml twice a day, it tells you about the volume of the mixture I am putting in the culture as the food. I took a look under the scope: the culuter appears to be more dense than it was last week.
<message edited by KathyL on Friday, August 26, 2011 5:11 AM>
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Re:Culture Journal, Species: Oxyrrhis marina
Saturday, August 27, 2011 5:47 PM
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Here is an un-concentrated squirt of 1 or 2 ml of my O marina culture in a 6 cm petrie dish, 40x thru the scope: click to see video.
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Re:Culture Journal, Species: Oxyrrhis marina
Monday, August 29, 2011 1:05 PM
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That's looking good, Kathy! Keep going!
--Andy, the bucket man. "Not to know the mandolin is to argue oneself unknown...." --Clara Lanza, 1886
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Re:Culture Journal, Species: Oxyrrhis marina
Tuesday, October 18, 2011 6:15 PM
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I checked my O. marina culture, and found out that it is mostly ciliates. I have been less than attentive to this culture, and haven't been filtering it as Andy does, so I got out my 37 micron filter and performed the appropriate maintanance on this jar. Lots of ciliates. It's amazing, when I try to culture ciliates, they crash, and when I try to culture something else, they contaminate and take over…. Back to twice a day feedings with Andy's mix and ChloramX.
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Re:Culture Journal, Species: Oxyrrhis marina
Tuesday, November 1, 2011 6:10 PM
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Still concerned that my O marina crashed. I haven't been adding any O marina from my culture to the copepod cultures. I kept a jar of something that I got by filtering one of my other cultures (flow thru) and I think its O marina. I added rotifers as a size reference. What do you think? Are the dots O. marina or ciliates?
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Re:Culture Journal, Species: Oxyrrhis marina
Tuesday, November 1, 2011 6:32 PM
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No clue without seeing them move, unfortunately. Seems like the right size.
--Andy, the bucket man. "Not to know the mandolin is to argue oneself unknown...." --Clara Lanza, 1886
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Re:Culture Journal, Species: Oxyrrhis marina
Sunday, November 13, 2011 7:05 AM
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i'm having trouble getting my video to upload to photo bucket. I'll try again later.
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Re:Culture Journal, Species: Oxyrrhis marina
Sunday, November 13, 2011 9:06 AM
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At long last: From my O. marina culture with an addition of rotifers, ciliates, and tigger poop from my mixed indoor culture, so you can acertain the relative size.
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Re:Culture Journal, Species: Oxyrrhis marina
Tuesday, December 20, 2011 12:10 PM
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O marina seems to have recovered from its ciliate contamination, and I'm now feeding it wild green phytoplankton of unknown species. (i also have a jar of O marina obtained from Glen King, who obtained it originally from me. It's good to have friends, and to give them some of your cultures.) I hope that it will convert the unknown fatty acids to DHA according to the paper I posted elsewhere, so the O. marina will be nutritious to the copepods. I shoud design an experiment to see if this is true. My wild phyto grows so much better than the Isochrysis.
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Re:Culture Journal, Species: Oxyrrhis marina
Tuesday, December 20, 2011 12:34 PM
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so happy it bounced back for you!
RLTW 180 Gallon Mixed Reef Then I heard the voice of the Lord saying, "Whom shall I send? And who will go for us?" And I said, "Here am I. Send me!" Isaiah 6:8
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