nannochloropsis culturing problems

[scooter413]

I am having a horrible problem culturing Nano. I am good for about 10 days then it turns brown and dies. Does the type of salt mix you use help or hinder algae growth. Right now I am using reef crystals and was wondering if it contains something the nano doesn't like.

Here is my procedure:

The well water Water is sterilzed. I have boiled it, I have used bleach then dechlorinator (sodium Thiosulfate), RO water before the Di resin and after the resin. (used one of each of these methods for at least two trials each )

All containers and air hoses are sterilized with bleach water and with alcohol

Florida aqua farms culture disk soaked in sterile water for 24 hours
added to 2 liters of boiled water(that's cooled down) with a salinity of 1.020
3ml of f/2 to 2 liters of water
filtered air (carbon and sterile cotton)
light from 2 brand new 2 bulb 4 ft fluorescent 6500 k bulbs (I have 8 one gallon glass jugs)(24hours)(The fixtures are old but bulbs are new)

After 7-8 days I add enough sterile water to make a gallon then continue with light and air
3 ml of f/2

(I also tried using 1 ml of f/2 in beginning and 1 ml at split with same results)

Day 10 dead.

This is getting expensive and time consuming for growing rotifers to feed to baby clownfish.

Questions:

The area that I am culturing is in the basement with a constant temp of about 75. Could the air have mold or bacteria in it that causes a problem?

Could older ballast not provide enough power to benefit from the 6500 k bulbs?

Is the salinity to high? (Trying to stay close to the feeding tank conditions)

Could my F/2 be contaminated?

Should I beat my head harder on the wall?

Any feedback would be greatly appreciated except the beating of my head part.

Scott

 

[diablomaster9045]

Here is how I do nano:

I am using 3 gallon carboys.

Using just RO/DI water and Fritz salt mix up a batch of saltwater to a salinity of 1.018. Add guillard's f/2 from Florida aqua farms in the amount directed on the side of the bottle. As for temp, my cultures sit at about 68 degrees.

Then add the culture vial and bubble vigorously but not cause it to foam up.

And wait for it to hit full density. Then when splitting the culture add 1/3 the recommended amount of f/2 and make sure you have a brewing hydrometer so you can easily measure the salinity. (Soak the hydrometer in alcohol between culture types.)

As for banging your head against something... A pillow is a much better substitute than a wall


Here is where I pick up my cultures from: (Hope I am not breaking rules by posting this link...)Algae | Carolina.com

 

[scooter413]

Thanks. A couple of questions.
Where would someone find Fritz salt. And are your carboys clear or the regular diffused plastic ones

 

[scooter413]

Never mind on the salt I found it.

 

[DFW]

I have been culturing nano for 3 years using a starter culture from a local reefer. I have had a few bottles crash, but have kept this strain alive, and never had every bottle crash in a batch. I rinse 2 liter bottles, and associated air tubing apparatus in hot water. Add approximately 1.019 sg fresh saltwater, 5 ml miracle grow plant food, 1 ml Kent essential elements, and enough nano to each 2 liter bottle to give the culture a green tint. I place 4 2 liter bottles like this in a 5 gallon bucket, attach air tubing to air pump, and place a 20 watt curly fluorescent bulb down the middle of the bottles. The bucket is lined with aluminum foil in hopes that it reflects the light around the bottles. Each day I twist the bottles a few times to get any sediment back up in solution. 2 1/2 to 3 weeks later the phyto is very dark green, and concentrated, and ready to go. During the warmer months it seems to be more likely to lose a bottle. I do this in a closed cabinet, so it gets a little warm. During the cooler months it seems to go especially well. I have used reef crystals primarily, and occasionally Instant ocean.

 

[scooter413]

I have not tried DFW's method yet, still trying to collect some 2 liter bottles. In the meantime I did some more testing and I have found that the ph of the cultures is running around 9.5 to 10.2. The crashed cultures are at 6.2. Would this high of a ph cause the culture to crash. Also the copepod tank that I have been trying to get established is also crashing and I checked the ph of that tank and it is at 4.5. Would the dead nano cause the ph to drop that low. The only thing that has been put into that tank is fresh water changes and cultured nano and nano paste(due to no fresh nano).

 

[DFW]

I never have checked the pH on my cultures. I know that now with the weather cooling down a bit, my phyto cultures are doing better. It won't take 2 1/2 weeks to complete a culture now. I've not cultured anything else.

 

[jservedio]

If you are going 8 days, it just sounds like you are letting your culture get way past max density and not splitting it in time so it is dying and causing all kinds of problems. With nano you don't have to be nearly as stringent with your disinfection unlike with something like iso. They key is just harvesting at the right time.

I had my cultures going for about 6 months using 2 gatorade bottles and a 23w CFL - I ended up dumping 95% of it out since i couldn't use it all or sell it. It's much better to have an under-dense culture than over dense. Never let it go too far before starting a new bottle.

 

[scooter413]

Never thought that I was going to long. A lot of post state they go up to two weeks. I guess that maybe it is maturing faster and choking itself out. I will try that. How many days do you let yours cook jservedio?

 

[DaveVS]

Any negative impacts to coral etc. from using Miracle Grow in your phyto?

 

[captainsaquarium]

I use miracle gro in my phyto and have had no issues.

 

[DaveVS]

Another question for those using miracle gro - do you dilute the miracle grow first (if so using indoor or outdoor dilution?) or do you add 5mL directly?