Hot!Phytoplankton Culture

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Fishtal
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Phytoplankton Culture

Culturing Live Phytoplankton
 
Microalgae is the base of the food chain and crucial to the captive breeding process.
    This is a simple guide to culturing your own live phytoplankton.  Live phytoplankton is useful if you are using the "Greenwater" technique for raising larval fishes.  It can also be used to feed rotifer cultures.
 

Things you'll need:
 
  • Live Phytoplankton Starter culture (You can get starter cultures from various sources such as Florida Aqua Farms, Reed Mariculture or even a local hobbyist)
  •  Flexible Airline tubing
  •  Rigid Airline tubing
  •  Air pump
  •  Culture containers (2L soda bottles work great. Drill a hole in the cap to accommodate the rigid airline tubing.  Collect some extra caps that remain undrilled.)
  •  Fertilizer (Some people use Miracle-Gro but I prefer Guillards F/2 solution from Florida Aqua Farms.)
  •  Clean culture medium (Saltwater with a specfic gravity of 1.015 with fertilizer added as per instructions.)
  • Light (a simple shop light or aquarium fixture is fine.)
  • Airline gang valves
  • Ordinary, household, 5.5% bleach
  • Dechlorinator (Some people use sodium thiosulfate for this, I use AquaSafe and it works fine.)
  •  Funnel
  • Plastic Bottle "crate" (The type that the soda bottles arrive at the store in.  I got mine from a local party store for free by simply asking.  These work great for holding the bottles... I've seen many people make elaborate contraptions to hold the bottles, these are perfect since that is what they were made for.)   
 

 

Getting Started: 
 
Sterilize the 2L bottles with bleach and dechlorinate. 
 
If you are using a commercially available starter culture simply follow the directions provided. 
 
If you are starting with a culture from a local hobbyist pour the culture into a 2L bottle and add culture medium until you achieve a light green color.
  • Insert a rigid airline through the hole in the cap, attach to a pump via flex airline and a valve, and start the air  flow, not too much.  Leave the cap on loosely so air can escape from the bottle.
  • Set in front of the light and wait a week. The culture will get darker, and in about a week's time, you will have a nice dense culture that can then be propagated by pouring a third of it into a prepared bottle as above.  The two thirds remaining can be used to feed your pods, clams, filter feeders, brine shrimp, rotifers, or fish larvae (via the rotifers).  More often, I split the dense culture in half by pouring half of the dense culture into a fresh bottle and refilling both bottles with the culture medium and starting over.  I do
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      Fishtal
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      Re:Phytoplankton Culture (permalink)
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      Edit:  I had these vids in the wrong thread.
      #2
      Evgen
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      Re:Phytoplankton Culture (permalink)
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      Hello Fishtal!
      In Your bottle is Nannochloropsis?
      You Use CO2 for high density of phito?
      #3
      Fishtal
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      Re:Phytoplankton Culture (permalink)
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      Evgen

      Hello Fishtal!
      In Your bottle is Nannochloropsis?
      You Use CO2 for high density of phito?


      Yes, I am culturing Nannochloropsis.
      I do not use C02, only air.
      #4
      Evgen
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      Re:Phytoplankton Culture (permalink)
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      only Nanno?
      I`m cultivated Tetraselmis, 2 week. 7 days - low green color.
      I use L1 medium DIY
      #5
      Fishtal
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      Re:Phytoplankton Culture (permalink)
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      Evgen

      only Nanno?
      I`m cultivated Tetraselmis, 2 week. 7 days - low green color.
      I use L1 medium DIY


      I do also culture Tetrraselmis but it is Nanno in the pics.  BTW, the rotifer videos belong in the rotifer thread, my mistake, they'll be moved.
       
      I'm not familiar with the L1 medium.
      #6
      Umm_fish?
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      Re:Phytoplankton Culture (permalink)
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      Here's my T-Iso setup:



      The bags hold around 6 liters when full.
      #7
      THEJRC
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      Re:Phytoplankton Culture (permalink)
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      How are you liking those bags Andy?

      I use "Crystal Geyser" natural spring water bottles (though I doubt the natural alpine water contents).  These jugs are nice square 1 gallon jugs that fit nicely on my culture rack.  I run 4 across on each shelf (4 ft shelf) with three twin tube NO Flourescents mounted vertically on the wall.  Using adjustable shelving each one of these units will give me between 12 and 16 culture spaes (3 or 4 shelves).  and run pretty efficiently.  The bottles come cheaply (10 for $10) and I utilize the water in other areas.  By using the disposable bottles I pretty much avoid having to clean vessels altogether.  A bottle will be used on average 3 times before being thrown away (unless it fouls sooner).  I replace the rigid on average every 12th cycle.

      On the subject of medium and Co2, I've bounced between a few mediums (and dependant on algae species will change) but have settled into the Guillards available in mass packs from FAF.  It's cheap and effective.  I run Full F strength on my Tetraselmis and Isochrysis, and just below full F on my Nannochloropsis.  I have long since removed my Co2 as I found it simply wasnt necessary for my cultures.  Split Nannochloropsis every 8 days, Isochrysis every 7th and Tetraselmis every 10th. 

      Co2 typically raises growth rate too high in my realm and I find that splitting too often not only stresses the cultures but leaves me with a ton of unuseable culture.  My goal is to maintain direct live sources and not have bottles of algae sitting in the fridge.

      Cultures are strained through a 250um seive for restarts / seeds, and a 20um sieve for harvests / feeds.  Occasionally I will "pelletize" samples prior to feeding using a microcentrifuge (this allows me to separate medium from the algal cells and will most likely prove to be more useful as my larvicology runs continue.).  The algae is centrifuged for approximately 2 minutes (or until visible separation occurs) the medium poured off, and the cells re-constituted using fresh saltwater.  I'm still working on this method to reduce cell damage but so far it's been an effective means of concentrating the cells in a cleaner culture (removal of the nutrients from the original medium).

      and thus ends my nifty brain dump!

      my racks look pretty much like this one (which sits behind my main system)


      Because it's all about the plankton!
      ~J      
      #8
      Umm_fish?
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      Re:Phytoplankton Culture (permalink)
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      I do like the bags, though I'm not generally a fan of plastic bags. But in this case since sterility is so important (as well as flow dynamics from crud buildup inside the container), I think they're good.

      What do you do to minimize contamination when transferring seed stock to a new culture? I have the narrow bottle neck as you can see. I'm using a sterile funnel, but flow rate is fairly slow and pouring through a funnel creates a suction that's taking any micro-organisms from the air straight into the culture.
      #9
      THEJRC
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      Re:Phytoplankton Culture (permalink)
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      I just pour direct through my strainer, while it's not the best approach I dont seem to have too many problems.  I do turn my lighting exhaust fans and such off prior to working with cultures    
      Because it's all about the plankton!
      ~J      
      #10
      Umm_fish?
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      Re:Phytoplankton Culture (permalink)
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      Okay, thanks. So I won't worry too much then.
      #11
      Gomojoe
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      Re:Phytoplankton Culture (permalink)
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      What size air pump do you guys use?  I have one from the skimmer to my nano that I never used.  It's got two ports and a dial, and says its rated at a pressure of .012MPa, and an output of 4.5L.min!  I'd like to get a setup that would do up to 7x 2L bottles and what I need for Rotifer cultures! My guess is I'd need alot bigger, but I just trying to get a reference.
      #12
      Fishtal
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      Re:Phytoplankton Culture (permalink)
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      I originally thought I'd need a bigger air pump too but I found that the cheap little ones do just fine. A couple of gang valves and some air line and you're all set.
      #13
      mickel
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      Re: Phytoplankton Culture (permalink)
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      Nice Post
      Mian ayaz
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