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Culture Journal, Species: Brachionus plicatilis
Thursday, August 11, 2011 6:55 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: Brachiionus plicatillis Species description: common L rotifer Culture source (link if possible ): Washington University Zebrafish Facility If algae, CCMP # (Optional ): http://ccmp.bigelow.edu/ Culture Establishment Date: 7/29/2011 , renewed by removing ciliates 8/15/2011 Continuation Date: forever if possible Culturing Vessel Details Salinity: 20 ppt Temperature: ambient, around 75-78F mostly pH: 8ish
Vessel description: 3 five-gallon buckets Lighting description: room ambient, fluorescent Lighting cycle: 14 day/10 night Aeration description: open rigid airline to bottoms of bucket. Fairly rigorous "boil" Methodologies Split methodology: Buckets are filled to about 4 gallons max. Bucket one is primary Culture. One gallon is dipped out daily from the Culture bucket and placed in the second, Enriching bucket, with some placed in the third Tiggerpods bucket. Fresh saltwater, one gallon, is dripped into the culture bucket during the day. Every week, rotifers and their culture water are decanted from the culture bucket into a clean bucket. Residual sediment is suspended in the residual water, and the remaining contents are poured via funnel into a 2 liter soda bottle and allowed to settle further. The original Culture bucket is wiped out with a paper towel, and the large volume of decanted rotifers and their water are returned to the bucket. This preserves any biofilm/filter that may have cultured on the bucket surfaces, while removing a majority of the decaying matter. After settlement, the soda bottle rotifers and their water are also decanted back into the Culture bucket. After about a month of this, the dripping method was abandoned. Too much work. Now I use 1-2 ml of RGcomplete or a mix of RG+, 1 part, plus 6 parts Nrich and an equivalent volume of 16 teaspoons chloramX/1 liter water. This is added twice a day. Once a day a gallon is dipped out of each bucket and and a gallon of 20ppt saltwater is added, room temperature all. I have one bucket of rotifers alone, mostly, and one bucket with every copepod in my basement included. I probably feed and exchange water out of the mixed bucket less than the rotifer only bucket, and yet they have equivalent or even more rotifers in the mixed critter bucket. Culture medium description: Chlorinated saltwater, obtained from spent water obtained from water changes of fish tanks, is diluted to about 20 ppt, and pH is adjusted. Bleach is added at 1 ml per gallon, and the water is allowed to sit overnight or longer. Water is dispensed into 1 gallon milk or other type of cap-able jug. Each day, one gallon is dechlorinated with excess ChloramX, and 6 parts N-rich with 1 part Rotigrow+ is added as needed. This medium is dripped into the culture bucket for several hours during the daytime. If more food is needed at night, a bolus of RotiGrow+ is added to the Culture bucket along with an = volume of ChloramX. Update: I've been bleaching and then the next day, dechlorinating (sodium thiosulfate) the whole 12 gallons or so at one time, and then using this water to do water changes on the rotifers, the copepods, the daphnia, the ciliates, and the dinoflagellates. ( Immediately after bleaching, I set up bottles for culturing phytoplankton, as the bleach will sterilze the bottles and airline sticks as well. I dechlorinate just before adding the f/2 and the innoculum.) Cell count: varies. I am trying an experiment to see how dense I can get a 4 gallon bucket culture . Once I started using the RGcomplete, my density increased visibly, though I have not done any serious counting. Reference links: Additional Information (No Pictures or Videos in the Section Please) Notes: If sediment is noticed in the Culture bucket during the week, a brine shrimp net is used to remove most of it. 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 Thursday, November 3, 2011 3:57 PM>
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Re:Culture Journal, Species: Brachionus plicatilis
Thursday, August 11, 2011 7:00 AM
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I need to keep better records of what I am doing. And be more consistent.
My goal with the record is to see how dense a culture I can obtain, given what I am willing to do to keep rotifers going. There may be a better way, but I am only willing to spend so much time and money on rotifers.
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Re:Culture Journal, Species: Brachionus plicatilis
Thursday, August 11, 2011 7:04 AM
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Today I checked my colony of rotifers. I doubt that I have even 50 rots per ml. However, I've been fairly consistent for the past couple of days with keeping the bucket tinted with food, and today I noticed for the first time that at least half of the rotifers are carying two eggs, instead of one or none. Seemed like there is food inside them, and I can expect some growth in the next 24 hours.
2 ml of RG+ in the media jug this morning. The drip is not perfect. Dripping can start at 2 drops per second, but as the hours go by, it slows down. I think the valve gets a little clogged. I've tried a few different valves, but nothing is ideal. May just have to start fast to end at the right rate. May just have to live with it.
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Re:Culture Journal, Species: Brachionus plicatilis
Thursday, August 11, 2011 7:11 AM
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great going!
I'm doing a bunch of nanno right now and in about a month I'll start my rotifers again. How do you like the RG+? Do you think it's more economic than nanno or just easier?
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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Re:Culture Journal, Species: Brachionus plicatilis
Thursday, August 11, 2011 7:26 AM
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easier and more effective. For clowns, no further enrichment is needed. I happen to have some N'Rich, that needs to get used, so I am enriching the rots further, but Roti-Grow + is really more than enough. It's no more expensive than nano or the old rotifer food that Reed offered.
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Re:Culture Journal, Species: Brachionus plicatilis
Thursday, August 11, 2011 7:19 PM
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This evening Culture looked good with many two egg females. Added 10 drop bolus Rotigrow+ and ChloramX.
<message edited by KathyL on Friday, August 12, 2011 5:59 AM>
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Re:Culture Journal, Species: Brachionus plicatilis
Thursday, August 11, 2011 11:21 PM
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Try bubbling your drip container with a little air to keep the RG+ in suspension. You'll get best results with multiple feedings if RG+ over the day. I'd try doing a drip in the AM and one late afternoon.
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Re:Culture Journal, Species: Brachionus plicatilis
Friday, August 12, 2011 6:00 AM
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Thanks, I'm trying that today.
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Re:Culture Journal, Species: Brachionus plicatilis
Friday, August 12, 2011 8:21 PM
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looks like I've got O marina in my rots….kinda messes with the experiment. I guess since my clownfish are past meta, I can just sieve the rots out, wash well, and start again………..It won't matter if it shocks the rot culture.
Well, I have to do that if I am to try this method with any kind of reliable, quantify-able results. OK, starting fresh tomorrow.
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Re:Culture Journal, Species: Brachionus plicatilis
Monday, August 15, 2011 11:15 PM
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Filtered the culture into a 53 micron. rinsed well, put into a gallon of clean saltwater, 25ppt in a clean bucket. Added 1 ml food: 6 parts N-rich to 1 part RG+. Added 1 ml food to another gallon of clean saltwater , with amply chloramX and dripped it in all day. Came home to a cleared culture, added 6 more drops food, 6 drops chloramX. Culture has had rigid airline with BOIL level of air.
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Re:Culture Journal, Species: Brachionus plicatilis
Thursday, August 18, 2011 6:01 PM
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August 17: Dripped another gallon with 2 ml food. August 18: Morning: About 10% of rots had 2 eggs, most had one egg. Removed one gallon to Enrichment bucket. Dripped in another gallon with 2 ml food over the day. came home to clear water, only half the gallon made it into the bucket. The siphon clogged. Loosened siphon, and water was tinted green by an hour later. Added food to enrichment bucket and Tigger bucket.
<message edited by KathyL on Thursday, August 18, 2011 6:58 PM>
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Re:Culture Journal, Species: Brachionus plicatilis
Saturday, August 20, 2011 6:26 PM
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Continued with above rhythm. Culture does not appear to be getting denser. Next, increase food to 3 ml per , twice daily. That O marina I had before turns out to be ciliates. Here's pix;  
<message edited by KathyL on Saturday, August 20, 2011 7:56 PM>
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Re:Culture Journal, Species: Brachionus plicatilis
Friday, August 26, 2011 4:22 PM
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I took a pudding cup's worth of culture, poured it thru a 53 micron screen, and back washed it onto a petrie dish( 6 cm diameter) . Here's a picture. Not many or any two egg rots, some one egg rots, and yet still lots o rots. 
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Re:Culture Journal, Species: Brachionus plicatilis
Saturday, September 3, 2011 8:52 PM
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I'm afraid I'm not being very consistent. I am still only on 2ml food twice a day, but I've switched back to just RotiGrow+ instead of the Andy recipe. I've wiped out the bucket, and split the culture into two buckets, as I ramp up in anticipation of having orchid dottybacks to feed. Of course that will be after MACNA, and someone will have to take care of the rots while I'm gone, and my favorite fish sitter will be at MACNA as well, so that leaves it to my darling daughter. I've given up on this drip idea as it is too fussy. Now I'm just shooting two ml RG+ in twice a day with chloramX. I did get an API ammonia test kit, and my ammnonia is between .25 and 0.5 mg/l, so that can be fixed with 2 ml of 120g/l chloramx. Right now my stock solution is 60 g/l, so I just add twice as much.
I do remove a third of the culture and replace it with 20ppt clean saltwater daily.
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Re:Culture Journal, Species: Brachionus plicatilis
Saturday, September 3, 2011 8:54 PM
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A quick check under the scope showed a lot of one egg rots, and quite a few 2 egg rots, so I think the change has done them no harm.
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Re:Culture Journal, Species: Brachionus plicatilis
Saturday, September 17, 2011 11:19 AM
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Before macna i got down to one bucket, and since coming back I've increased feeding and now have a pretty dense looking population. 4 gallons, one gallon removed per day to either feed larvae or something else. replaced with 20 ppm clean water daily. chloramX, and rotigrow + ...@ two ml twice a day. Culture seems to be thriving, and doing better than the slow drip. Every few days, I refresh the bucket to get rid of detritus. As of 9/26/2011. Now I'm sure I've jinxed it.
<message edited by KathyL on Monday, September 26, 2011 8:28 PM>
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Re:Culture Journal, Species: Brachionus plicatilis
Monday, October 3, 2011 7:21 AM
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RGcomplete arrived Friday. It's Monday. I've used it at 3 times the volume I was feeding RotiGrow + for my rotifers, as directed. This morning, there are significantly more rots in the filter, and when checked under the scope there are quite a few 2 egg females, whereas it has been quite a while since I saw any 2 egg females under the old regimen. I was skeptical at first, but the conversion is happening. This may be a great product. Thanks to the Reeds and Gresham.
<message edited by KathyL on Wednesday, October 5, 2011 8:51 PM>
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Re:Culture Journal, Species: Brachionus plicatilis
Wednesday, October 5, 2011 4:34 PM
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The rotifers and I are loving RGcomplete. The indoor polyglot culture returned a dense brown, critter concentrate from a gallon of culture, once filtered. Daphnia, T. californicus, and rotifers are thriving.
<message edited by KathyL on Wednesday, October 5, 2011 6:23 PM>
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Re:Culture Journal, Species: Brachionus plicatilis
Wednesday, October 5, 2011 6:16 PM
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RGcomplete... one word (just like the label)  I'll hound you all till you get it right. I will not have another Tiger-pod deal on my hands
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Re:Culture Journal, Species: Brachionus plicatilis
Wednesday, October 5, 2011 6:21 PM
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whatever we call it, surely you cannot miss the praise this product is receiving. When will it be commercially available?
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