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.
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<message edited by KathyL on Thursday, November 3, 2011 3:57 PM>