MBI Event Reporting Form

GENERAL
Name:	Jeremy Neff ( JNeff ) Site: Marine Breeding Initiative (MBI)
Species:	Nannochloropsis Sp. ( Class: FC1 , Points: 5 )
Establishment Date:	08/01/2010
Culture Source:	http://florida-aqua-farms.com/
Link to Journal:	http://www.mbisite.org/Forums/tm.aspx?high=&m=94979&mpage=1#94980

Culturing Vessel Details
Temperature:	75-80 ° F
PH:	8.1
Specific Gravity:	1.012
Vessel Description:	I make larger batches in a white 5 gallon bucket and smaller batches in clear 2 liter bottles.
Lighting:	I use a 10 watt CFL suspended 5" above a white 5 gallon bucket or two 48" fluorescent plant grow bulbs in a shop light fixture suspended 5" above a row of clear plastic 2 liter bottles sitting on top of white laminated particle board.
Lighting Schedule:	24 hours continuous lighting

Methodologies
Feeding Tactics/Medium:	I add 1.45 ml/gal Florida Aqua Farms Inc. Micro Algae Grow liquid as fertilizer into dechlorinated salted tap water. The salt is Instant Ocean brand. The specific gravity is 1.012. This culture medium is aerated 24 hours before I add the green water inoculation.
Split Cycle:	All culture water is inoculated with roughly 1 part green water to 10 parts culture water. If I need green water in a hurry this can be increased to a 50/50 split. I allow newly inoculated culture water batches to "green up" for 5 days to a light to medium green. After 5 days I take #2 two liter bottles worth out to store in the refrigerator for future batch inoculations (these remain viable for 2-3 months if you shake them regularly while in the fridge). The remainder of the batch "greens up" for another 2-5 days. Usually, by days 7-10 it is ready to harvest when dark green.
Split Yield:	When I culture in 2 liter bottles it is a straight batch culture with all of the cells being harvested and either fed off or stored in the refrigerator for future culture inoculations. When i culture in a 5 gallon bucket I remove 4 liters of water at day 5 to store for future cultures. The remainder of the 5 gallon culture is then allowed to fully mature and be harvested for immediate use.
Number of Cultures:	1-3 actively growing cultures and 2 dormant refrigerated cultures
Number of Splits:	Starting 2010 with an average of 2 splits a month = roughly 132 splits
Density:	not measured
Harvest Methology:	When I culture in 2 liter bottles the entire bottle is capped and put in the refrigerator when mature or immediately used if needed. When I culture in a 5 gallon bucket I will dip out a cup at a time to feed rotifer cultures. I also utilize the greenwater technique for larval rearing. I pour greenwater directly from the 5 gallon bucket into the hatch tank to green up the water.
Adjustments Made:	In the past I set the SG at 1.025, but I have found that nannochloropsis oculata (and brachionus plicatilis) multiplies faster at a lower salinity. I now prep my culture water with a SG of 1.012.

Other Details
Other Information:	The reason I use the specific gravity of 1.012 is because I have found that my L strain brachionus plicatilis multiply rapidly at that salinity. I can add a small amount of rotifer culture into a large amount of green water when I want to maintain rotifers but am not actively feeding with them. If you do this when the salinity of the greenwater is different from the rotifer culture they don't usually survive. I find it more convenient to add a small amount of rotifers and not have to touch it for a week while they multiply and exaust the greenwater vs. feeding a small amount of greenwater to the rotifer culture daily.

Other Details

Other Information:

The reason I use the specific gravity of 1.012 is because I have found that my L strain brachionus plicatilis multiply rapidly at that salinity. I can add a small amount of rotifer culture into a large amount of green water when I want to maintain rotifers but am not actively feeding with them. If you do this when the salinity of the greenwater is different from the rotifer culture they don't usually survive. I find it more convenient to add a small amount of rotifers and not have to touch it for a week while they multiply and exaust the greenwater vs. feeding a small amount of greenwater to the rotifer culture daily.

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