phyto culturing newb...

[OG-Goby]

i looked, but didn't find a dedicated forum for phyto... if it exists, someone point me to it!

I started a culture last night, but I am thinking I must have done something wrong. I bleached all equipment in a 10% solution for 10 mins. rinsed in RO water. then used alcohol to sterilize the jar, tube, lid, and bulkhead for the lid wearing nitrile gloves the entire time. I prepped a solution of 1 liter fresh made saltwater with .25ml of f/2 and boiled in an electric kettle. afterwards I put in sterile jar from before and put it in the fridge until it cooled off. in the meantime I put my jar of phyto on the counter (tetraselmis) to warm up some. in the end, I added 12oz of saltwater and fert, and 12oz of tetraselmis culture from TSA (who states its culture ready). today, the jar seems to have a film of green on the jar, that does not disperse if i agitate it some... i think my air pump was dialed too low, it was bubbling, but it seemed like the tetraselmis was not fully in motion in the jar, so i opened up the valve some.

I realize its probably newb stress, but hoping someone can tell me if i went wrong and how to correct? or if i need to just dump this round and try again?

also - anyone that is sterilizing via boiling, what do you do with the water while its cooling to keep it sterile?

thanks in advance. Im probably over-reacting...

 

[The_Paradox]

No need to boil. What salinity and what brand f2? I ask because some vendors I need 1ml/L others I need to to 4ml/L. Get aeration up for sure. To verify if the culture is still viable just check it under a scope. Tetra is motile so easy to tell. Will let you know if the culture is shot or possibly contaminated with nano or something else non-motile.

 

[OG-Goby]

salinity was 35ppt. (as per TSA they culture at NSW). the f/2 was from Aqua Algae on Amazon and the bottle states its a copy of Guillards f/2 and to use 1ml per liter... oh... yeah, ok... i need more. I went with the 1ml per gallon but didnt read the actual bottle. doh.. i only added .25ml to the liter as I thought 1ml per gallon. so I assume i need to add maybe .5ml as i have a total of 24oz at the moment, and it does have a very small amount in it already.

havent bought the scope yet... guess I need to get on that.

 

[The_Paradox]

salinity was 35ppt. (as per TSA they culture at NSW). the f/2 was from Aqua Algae on Amazon and the bottle states its a copy of Guillards f/2 and to use 1ml per liter... oh... yeah, ok... i need more. I went with the 1ml per gallon but didnt read the actual bottle. doh.. i only added .25ml to the liter as I thought 1ml per gallon. so I assume i need to add maybe .5ml as i have a total of 24oz at the moment, and it does have a very small amount in it already.

havent bought the scope yet... guess I need to get on that.

Dose up for sure. Also would drop the salinity to 30ppt.

 

[Pistondog]

A couple of bubbles per second. Your 1 to 1 water to phyto ratio should be ok, i never went higner than 4 to 1. But too dilute is not good, the goldilocks concentration to outcompete other stuf, but not interfere with growrh.
Never sterilized water, just vessels with alcohol overnight.
Good luck, another fun aspect of this hobby.

 

[OG-Goby]

A couple of bubbles per second. Your 1 to 1 water to phyto ratio should be ok, i never went higner than 4 to 1. But too dilute is not good, the goldilocks concentration to outcompete other stuf, but not interfere with growrh.
Never sterilized water, just vessels with alcohol overnight.
Good luck, another fun aspect of this hobby.

I went 1:1 as that seemed to be the general rule to follow. End goal is half gallon so when this 32oz container is mature I’ll do 1:1 in a half gallon mason jar.

I’m sterilizing the water as “practice” for isochrisis. The end goal is one green one gold to dose the display tank.

Thanks for the reply. At this point I’ll take any advice I can get!

 

[OG-Goby]

Dose up for sure. Also would drop the salinity to 30ppt.

Done and done. Let’s see how it looks tomorrow!

I boiled the water and fert as practice for when this tetraselmis is working I want to try isochrisis. So I am trying to find the best practices for sterilization while it’s not as important to figure out my routine.

Do you do any cultures that are this picky? I did order some sodium thiosulfate so I can bleaching and dechlor a try as well, but again not sure how to keep that sterile while bubbling the dechlor. Seems like a potential contaminant window. But again, total newb here so I am over thinking 99% of this.

 

[Pistondog]

I went 1:1 as that seemed to be the general rule to follow. End goal is half gallon so when this 32oz container is mature I’ll do 1:1 in a half gallon mason jar.

I’m sterilizing the water as “practice” for isochrisis. The end goal is one green one gold to dose the display tank.

Thanks for the reply. At this point I’ll take any advice I can get!

Iso likes it more dilute to start, and lower light levels to start. I put a piece of copy paper at the vessel, 'blocking' the light. After the culture started to darken, remove the paper.