Pod Culture Questions

[pmcmahon]

So I've started learning to culture pods and phyto as a first step in possibly trying to raise fish. For the moment I'm just experimenting and whatever pods I culture I put into my refugium tanks as I set up for my next tank. I started with Tig, Tisbee, and Apocalypse pods. I also found a local reefer to pick up a phyto culture that was I think originally Algae Bard Ocean magic. I looked at it under my (very basic) microscope and it appeared to be largely one species dominant at that point. My guess was nanno.

So the results of this little experiment were that my Tig pods got in everything else. I had trouble keeping the Tisbee pods and they seemed to be dwindling down instead of getting more dense. My phyto I think has devolved into a pretty generic "green water" at this point that I'm reasonably sure is contaminated with something that I have to keep fighting because it forms a film next to my light source. Not bad all things considered. I dumped everything but the Tig's into the fuge and keep the phyto going to feed them. I think it's time to try again with some fresh cultures. Goal one is to see if I can keep some single species cultures of phyto for a few generations. Goal two is to see if I can have more success with the different types of pods and hopefully avoid cross-contamination this time. I've watched a few of Todd Gardner's videos and I think I'm ready for attempt number two.

So here is where I have a couple of questions for the group:
1: Do you guys keep sterile primary cultures of phyto and bloom a secondary from that? Tips?
2: How do you guys clean the crud out of your pod culture tanks? I've seen people just suck it out with a turkey baster but it seems like that's a huge portion of the culture you lose just to try and clean the mulm
3: My tig culture vessels keep getting what looks to me like mineral deposits (picture attached) on the airlines and side of the container. Can anyone confirm and help with how to avoid this?

Also attached is a picture of my setup. Please let me know if you see something I could improve on!

 

[HankstankXXXL750]

I started doing this and have had some success. I got my starter cultures from @Eldredge here on R2R and he gave me great advice on how to proceed.

You cannot culture a mix like ocean magic as one phyto will out perform the others. Have to run individual cultures. I am currently running 5, but a couple crashes and am restarting. I have two sets of buckets and have the second set cleaned, sterilized and refilled with sterilized saltwater (RODI/Instant Ocean/Bleach) day before I switch I dose ascorbic acid to neutralize the bleach. Then I drain 1/2 the new water into buckets (I fill them all the way so the bleach water helps to keep it all sterilized until I’m ready) then I transfer 1/2 of the culture into the new bucket and add F/2 and run. Then the other 1/2 I mix together in equal parts then pour together into storage containers that I refrigerate while using it. I dose my tanks and my pods.

for my pods I run 1 gallon glass jars and when I harvest I suck out the gunk and everything from the bottom and then pour through a series of sieves to separate. I may end up with a portion of the “gunk” in my harvest but mostly it is clean. I have sieves from 800,250,120,52 micron.

when I first started I pulled a 16 Oz bottle of each and kept open for air and sealed and inverted everyday for a back up starter. Once I moved from the 1 gallon jugs to the 5 I quit doing that (wish I hadn’t as I had to order replacement starters from Dave when they crashed this week lol). With starting transferring each week I will probably either keep back a portion or start running my 1 gallon system as a back up to my 5 gallon system.

as far as the build up on the pod vessel are you using RODI?

 

[DaJMasta]

Single strain algae cultures are the way to go, you keep the variety without losing some to outcompetition, and it's much easier to see if they are contaminated/going bad. I like to just pour 1/6th or so of the culture into a new jug at harvest, dilute with fresh saltwater, and fertilize - then your new culture is started and the remaining 5/6ths of the old culture can be fed.

As for the copepods, my experience is that apocyclops is the one of the three that's easiest - it will handle almost as much feeding as you can get it (I was feeding something like 1/10th of the culture daily and refilling with only phyto and it was dense and happy), whereas both tigriopus and tisbe both want clearer water - too heavy feeding and the cultures will slowly crash. I aim for 2:1 to 3:1 fresh saltwater to phyto culture when feeding both, but basically, if the water looks tinted at time of harvest, I just don't add phyto and only replace with fresh water that day. I'd say apocyclops likes fairly high aeration (to support high culture density) while I use pretty low aeration for tisbe and tigriopus.

If you're harvesting every day and either feeding or just replacing the water, the cultures basically don't need to be cleaned, in my experience. I do feed from a spout at the bottom, which helps flush out grime, but I will go months (sometimes more than half a year) before it looks like there's enough crud in there to transfer them to a new vessel and clean the old one out.

 

[LordofCinder]

I clean my phyto tools wit vinegar before using to disinfect it. Best to try to culture one species of phyto only, I've never had luck with the mixed phyto's. I currently culture nanno and tetra, they are both easy.

I let the pods culture and then harvest them 100% every 3-4 weeks, then I completely clean the container, disinfect it, and use a little of the pods to start the new culture. I don't worry about the crud buildup because I completely start over every month.

I use tupperware containers from the dollar store for my pods, I currently culture tisbe and apocyclops. I gave up on tiggers, they are big, ugly, and I never see them in the tank, I don't believe they do well in reef tanks.

I don't know what is that mineral buildup is.

Its all a learning process, you will learn more and get better every time you do it

 

[pmcmahon]

I have two sets of buckets and have the second set cleaned, sterilized and refilled with sterilized saltwater (RODI/Instant Ocean/Bleach) day before I switch I dose ascorbic acid to neutralize the bleach.

I've never heard of this technique. Do you have details on how much bleach you dose per gallon or 5? Is there a calculation you do to match the ascorbic acid to neutralize? I haven't been sterilizing my saltwater because I didn't know how to get that done.

as far as the build up on the pod vessel are you using RODI?

Yes, all RODI and fresh mixed salt. It looks like calcification or something similar-ish to hard water mineral deposits. I was wondering if the phyto changes the PH enough to start pulling some of the minerals out of the salt turning it into a solid that adheres to the vessel. Maybe I should test the PH of my culture. Thoughts?

 

[pmcmahon]

As for the copepods, my experience is that apocyclops is the one of the three that's easiest - it will handle almost as much feeding as you can get it (I was feeding something like 1/10th of the culture daily and refilling with only phyto and it was dense and happy), whereas both tigriopus and tisbe both want clearer water - too heavy feeding and the cultures will slowly crash.

Thanks for the reply. Do you only feed phyto to your cultures or do you supplement with crushed pellet or flake on any of them? What strain or strains of phyto do you feed?

 

[pmcmahon]

Sorry, I've been slow to respond to this thread. Work got crazy and then I was on vacation. I appreciate everyone's advice. A few more follow-up questions.

1. When you guys add F2 do you try and sterilize it at all or include it in your sterilization process?
2. I've just started out my cultures by dosing whatever the recommended dose on my F2 bottle (Fritz 2 part pro) is at the beginning of the culture. Do you guys monitor nitrate levels in the culture and continue to dose until the culture is up to the required density or just put in one really large dose at the beginning and let it ride?

 

[DaJMasta]

I don't use foods other than phyto - I had some luck supplementing with other powder foods, but it involves another step and thing to keep track of, and I didn't see any better results. I think once a culture is established, the bacteria and whatnot in there supplement the phyto addition. It's also easier to gauge how much food went in when it's phyto, so you run less risk of overfeeding and not realizing it.

As for the phyto fertilizer, I don't do anything special aside from careful handling of the bottles. I make sure to use a clean pipette to get it out of the container and I make sure that if the pipette touches the side of a jug with a phyto culture in it that I immediately replace it to prevent back contaminating the fertilizer. Cool storage also limits growth for the fertilizer, but I don't refrigerate or anything. I've seen some kind of mold growing on the paper seal top of one my big fertilizer bottles, but I haven't noticed an effect.

I believe proper lab technique would be to put the fertilizer and seawater in a flask, then cover with tin foil and stick the whole thing in an autoclave, cooling and then uncovering only for innoculation. The trouble here is that an autoclave isn't a normal household device, and there are least some components of f/2 that will denature with heating, so you can't exactly ensure that it's sterile without risking cooking it and changing some components. In a pinch, you could probably double boil the bottle (opening to remove expanding air as it heats), but it's not a controlled method.

I don't test water in cultures - while annoyance and low maintenance is a good enough reason to not to outside of maybe commercial/lab settings, I think the risk of cross contamination would go up 10x or more by testing mid cycle even once - I don't think it's worth it. Then there's the fact that microorganims are generally more tolerant of nitrates and such, and that phyto actually likes it, so much, much higher than normal levels would be required to upset it. If I were going to test it at any point, I would test nitrates at the end, as this would give you an idea of fertilizer consumption, and would give you some assurance that you weren't then dosing nitrates with your fertilizer - but if you start low and ramp up the amount until the container goes basically opaque for your culture conditions, that's probably about the point where they run the fertilizer out too.

 

[PotatoPig]

Cheat Mode: Get some spirulina powder and mix a pinch in with RODI and feed that to your pods.

Cheap. Easy. Effectively infinite shelf life. You can convert your phyto culture vessels into more pod cultures.