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HOW TO: Clean, easy and highly nutritious greenwater culture for Daphnia and Moina.

dw1305

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Hi all,
I was hoping @dw1305 would have a theory as our resident bacteria and water quality expert?
I'm afraid not.
It doesn’t have any odour as far as I can tell. I’ll just have to keep an eye on it, if the population seems to decline, I’ll do another water change.
I'd probably go down that route as well.

cheers Darrel
 

Wookii

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So my culture has redeveloped the dreaded green/yellow clouding, and I noticed a definite drop off in Moina population. More alarmingly I noticed loads of the snails clamouring above the water line.

So I did another 95% water change, but also retained a sample of the old water to test for ammonia and nitrite. Sure enough, the ammonia level was quite high, and the nitrite was a definite non-zero value also.

There are no obvious dead snails, so I’ve assumed the source is some of the old Catappa leaves I transferred from my main tank - though I was under the impression they had no organic content that could foul water once dried.

The fact that there was a fair bit of nitrite present actually might suggest that the sponge filter is active, and starting to process any ammonia, so hopefully it will start to deal with the biological filtration for the tank proper.

In the meantime, I’ll keep on with regular water changes.
 

louis_last

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The moina in my main culture have actually now been overtaken by the ceriodaphnia, I think there may actually be hundreds of thousands of them in there and I'm literally throwing ceriodaphnia away each day to keep the population under control because my fish can't eat this much. It's frustrating because I've never been able to get them to these numbers in another culture and I don't know what it is that I've done right with this one.
There are no obvious dead snails, so I’ve assumed the source is some of the old Catappa leaves I transferred from my main tank - though I was under the impression they had no organic content that could foul water once dried.

Surely it's the fish mix?
 

Wookii

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The moina in my main culture have actually now been overtaken by the ceriodaphnia, I think there may actually be hundreds of thousands of them in there and I'm literally throwing ceriodaphnia away each day to keep the population under control because my fish can't eat this much. It's frustrating because I've never been able to get them to these numbers in another culture and I don't know what it is that I've done right with this one.

That is a shame. I might have to have some more off you once I sort my current problem out, and just have them in with the moina.

Surely it's the fish mix?

Of course, you're right, I hadn't thought of that. I tested my current Chlorella culture mix that I thought was 'fully' mature and its got plenty of ammonia in it.

So am I basically feeding the chlorella too early? Is it basically ready to feed once all the ammonia is gone?

This might actually spur me on to try a dry salts fertilised bottle bottle next, as then it should be ammonia free throughout.
 

louis_last

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That is a shame. I might have to have some more off you once I sort my current problem out, and just have them in with the moina.



Of course, you're right, I hadn't thought of that. I tested my current Chlorella culture mix that I thought was 'fully' mature and its got plenty of ammonia in it.

So am I basically feeding the chlorella too early? Is it basically ready to feed once all the ammonia is gone?

This might actually spur me on to try a dry salts fertilised bottle bottle next, as then it should be ammonia free throughout.
I'm really interested in what results you can get with dry ferts because I genuinely can't tell whether it will produce better or worse results, seems like it could go either way, but if it simplifies things in any way you wont have to twist my arm to get me to switch over.
There's a very noticeable change in smell once all the fish mix has been consumed I find, it stinks when I first run air through it but by the time it's all used up there really is no smell other than a faint grassiness from the chlorella. If you turn off the airstone too and let it settle a bit you can normally get a sense of whether there's any browning left from the fish mix in the water around the edges. Once the bottles have gone fully green and opaque if you leave them for a couple more days they just get even darker, the 5L ones almost look black because the cell density is so high and if I shine a seriously bright LED torch through them there's not even a hint of light penetrating through to the other side. It's complicated by the fact that we're all using slightly different emperatures, amounts of light, vessel size/shape etc.
One of the videos from malaysia I found talking about how they grow chlorella actually described using ammonia and said that the chlorella was ready to use once it no longer smelled of urine. Are you shaking the fish mix before you use it to make sure it's not settling into layers or anything weird like that?
What photoperiod are you using for the chlorella? I was running 24 hours of LED at one point but I actually reduced it down to 18 after something I think you linked to pointed that out as the optimum photoperiod. It may be that the the chlorella only uses up certain components of the fish mix in the dark too because I seem to be getting better results since.
Please keep us posted and consider experimenting with the dry ferts on all of our behalf. No problem at all to send you some more ceriodaphnia to add in with the moina, I'll try and get some over to you this week.
 

Wookii

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I'm really interested in what results you can get with dry ferts because I genuinely can't tell whether it will produce better or worse results, seems like it could go either way, but if it simplifies things in any way you wont have to twist my arm to get me to switch over.
There's a very noticeable change in smell once all the fish mix has been consumed I find, it stinks when I first run air through it but by the time it's all used up there really is no smell other than a faint grassiness from the chlorella. If you turn off the airstone too and let it settle a bit you can normally get a sense of whether there's any browning left from the fish mix in the water around the edges. Once the bottles have gone fully green and opaque if you leave them for a couple more days they just get even darker, the 5L ones almost look black because the cell density is so high and if I shine a seriously bright LED torch through them there's not even a hint of light penetrating through to the other side. It's complicated by the fact that we're all using slightly different emperatures, amounts of light, vessel size/shape etc.
One of the videos from malaysia I found talking about how they grow chlorella actually described using ammonia and said that the chlorella was ready to use once it no longer smelled of urine. Are you shaking the fish mix before you use it to make sure it's not settling into layers or anything weird like that?
What photoperiod are you using for the chlorella? I was running 24 hours of LED at one point but I actually reduced it down to 18 after something I think you linked to pointed that out as the optimum photoperiod. It may be that the the chlorella only uses up certain components of the fish mix in the dark too because I seem to be getting better results since.
Please keep us posted and consider experimenting with the dry ferts on all of our behalf. No problem at all to send you some more ceriodaphnia to add in with the moina, I'll try and get some over to you this week.

Thanks @louis_last - I don't find the fish mix all that pungent, maybe because it reminds me of ground bait additives I used to use when I used to go fishing - so I get pleasant subconscious flashbacks lol. Some of the fish oil pellet dips I used to use when barbel fishing could blow your head off. I will do a sniff test on the Chlorella culture anyway - I've got it up high on a storage unit, so it takes a bit of effort.

I suspect I'm not getting enough light into my Chlorella cultures as they're taking too long to green up, so I may have to source some shallower and wider clear containers - my current ones are very narrow and deep. So that may be why, after two weeks, I've still got ammonia in the culture. I run the lights for about 16 hours currently. I do shake the fish mix vigorously before adding to the water though, so I don't think that's an issue..

I will make the next culture a dry salts one then, and see how it goes. My one concern is how I add the necessary vitamins in - any suggestions? Also, what should I do about the sugar element. Is it really necessary, and if so should I add glucose in lieu of molasses, just stump up and get molasses?

Any suggestions for a dry ferts mix, and dosage level? I'll probably make up a bottle of all in one solution, but I have all the necessary dry salts (KNO3, MgSO4, K2SO4, KH2PO4) and APFUK Micro mix.

Thanks for the offer of the ceriodaphnia - I'll hold off though until I definitely have things running smoothly with the current Moina culture, as it's just not working right at the minute. I may have to switch back to yeast to maintain the current culture until the Chlorella is ammonia free.
 

louis_last

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I don't think that the sugars from the fishmix are strictly necessary, it just allows the chlorella to continue growing during the dark period and they are included with a view to promoting microbial growth around the root system of terrestrial plants more than for their value as a fertiliser. It just happens that Chlorella can be grown mixotrophically and exploit the sugars directly.
I have literally zero experience with dry salts so I can't offer any useful advice there. I wish I could test my chlorella culture to see what the levels are like over time, what sort of tests are you using? are they expensive?
If there's something off about the amount of light and the size and shape of your containers is it also possible that some of the chlorella is dying in the actual chlorella culture before it even reaches the moina somehow and producing ammonia?
I wonder if @MirandaB has experienced anything similar with the bottles she's still growing on the windowsill?
One of my backup cultures in a plastic pint glass got fed with a huge amount of chlorella and has been totally neglected since then with no water changes and has good population growth and no signs of a crash so it seems it must be possible under some conditions to purge any harmful ammonia or nitrite.
I wonder about a drop or two of seachem prime in the chlorella before feeding as a safety measure maybe?
 

Wookii

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I don't think that the sugars from the fishmix are strictly necessary, it just allows the chlorella to continue growing during the dark period and they are included with a view to promoting microbial growth around the root system of terrestrial plants more than for their value as a fertiliser. It just happens that Chlorella can be grown mixotrophically and exploit the sugars directly.
I have literally zero experience with dry salts so I can't offer any useful advice there. I wish I could test my chlorella culture to see what the levels are like over time, what sort of tests are you using? are they expensive?
If there's something off about the amount of light and the size and shape of your containers is it also possible that some of the chlorella is dying in the actual chlorella culture before it even reaches the moina somehow and producing ammonia?
I wonder if @MirandaB has experienced anything similar with the bottles she's still growing on the windowsill?
One of my backup cultures in a plastic pint glass got fed with a huge amount of chlorella and has been totally neglected since then with no water changes and has good population growth and no signs of a crash so it seems it must be possible under some conditions to purge any harmful ammonia or nitrite.
I wonder about a drop or two of seachem prime in the chlorella before feeding as a safety measure maybe?

I've only tested for ammonia and nitrite, since they are the harmful ones, and just tested with basic API aquarium test kits.

What size and shape container are you using? If it is actually chlorella dying and causing ammonia then that will be a pain, as I wouldn't know how to avoid that. It seems unlikely, but I have no way of knowing, other than if I can culture with dry salts which I know wont contain any ammonia.

I am going to try simply culturing in some clear mineral water bottle on the window sill too like @MirandaB, though I don't have a south facing window, so they'll only get direct sunlight for a proportion of the day.

Seachem Prime only binds ammonia for up to 48 hours, so I don't think that will be a viable solution.

For the dry salts mix then, I will just try the 3 : 3 : 1 mix of N : P : K recommend in that white paper I linked to previously, or at least I'll add the same proportions of N and P, the K will have to be whatever comes out since KNO3 and KH2PO4 both contain K. I think the article suggests 100mg/litre of N from what I can decipher (my chemistry isn't that good).
 

MirandaB

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Not had any problems using the fish mix fed Chlorella as yet although I had another die off of Moina which wasn't down to that I believe as water tested fine.
Went out with the torch at night and found what appear to be Moina under the microscope in one of my tubs so collected a load popped them in the tank and they're booming.
I tend to not dump too much in at once so feeding daily as they clear it in 24 hours,I also keep floating plants in the tank too to help with water quality.
Also running a back up tray of Chlorella outside,that's just a large shallow cat litter tray and I've chucked a load of Tomato fertilizer I found in the shed in there to see how that works.
 

louis_last

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I've only tested for ammonia and nitrite, since they are the harmful ones, and just tested with basic API aquarium test kits.

What size and shape container are you using? If it is actually chlorella dying and causing ammonia then that will be a pain, as I wouldn't know how to avoid that. It seems unlikely, but I have no way of knowing, other than if I can culture with dry salts which I know wont contain any ammonia.
I've only ever used 2L and 5L bottles of coop own brand still water. At first I even used the water as a base for the fert mix but now obviously I'm using tap water without any problems. I use an arcadia jungle dawn LED bar upright and aligned with the bottles now for maximum light and it's 34w, not 34w equivalent, but actual 34w.
I've been looking very closely at all my cultures based on what you've been reporting here and I actually am experiencing something similar in some of them because some but not all of the pint glass backups and test cultures that have been fed with chlorella have a very brown tint to them even after the water has cleared. It's not quite brown like the fresh fish mix, more like tannins because the water is very clear, but in the smaller cultures it can't be - there's no leaves in the water and they weren't made with tank water that has tannins or rooibos added to it.
I just did a huge water change on the master culture because it was due anyway and looking at the water coming out it really is pretty brown. I've not had any crashes for a long time and previously I even mentioned here that the water in my culture was quite tannic but now I'm wondering whether sometimes there is something being left behind by the fish mix. I'm keen to see your results with the dry ferts but it may just be the case that sometimes we're using slightly too much fish mix?
The dosage I initially went with was just pulled straight out of my ass, it wasn't based on any real expertise - just a lockdown obsession with moina and chlorella.
The first time I ran the 5L bottles I actually used a stronger mix than usual (7.5ml fishmix as opposed to 5ml) and it didn't seem to make any difference, if anything there was more clumping and sedimentation when they were getting started. The most recent batch I used less than 5ml and it hasn't had any obvious negative effect, they may even have colonised slightly faster.
I've spent an awful lot of time tinkering about with this and I regret not being slightly more disciplined and scientific in my approach now.
Provisionally it seems the safest thing to do until we can work out whether dry ferts are a better option is just to reduce the amount of fishmix by half. I don't believe it'll have a detrimental effect on how fast the algae grows, maybe only the ultimate cell density of the cultures.
 

Wookii

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I've only ever used 2L and 5L bottles of coop own brand still water. At first I even used the water as a base for the fert mix but now obviously I'm using tap water without any problems. I use an arcadia jungle dawn LED bar upright and aligned with the bottles now for maximum light and it's 34w, not 34w equivalent, but actual 34w.
I've been looking very closely at all my cultures based on what you've been reporting here and I actually am experiencing something similar in some of them because some but not all of the pint glass backups and test cultures that have been fed with chlorella have a very brown tint to them even after the water has cleared. It's not quite brown like the fresh fish mix, more like tannins because the water is very clear, but in the smaller cultures it can't be - there's no leaves in the water and they weren't made with tank water that has tannins or rooibos added to it.
I just did a huge water change on the master culture because it was due anyway and looking at the water coming out it really is pretty brown. I've not had any crashes for a long time and previously I even mentioned here that the water in my culture was quite tannic but now I'm wondering whether sometimes there is something being left behind by the fish mix. I'm keen to see your results with the dry ferts but it may just be the case that sometimes we're using slightly too much fish mix?

The dosage I initially went with was just pulled straight out of my ass, it wasn't based on any real expertise - just a lockdown obsession with moina and chlorella.
The first time I ran the 5L bottles I actually used a stronger mix than usual (7.5ml fishmix as opposed to 5ml) and it didn't seem to make any difference, if anything there was more clumping and sedimentation when they were getting started. The most recent batch I used less than 5ml and it hasn't had any obvious negative effect, they may even have colonised slightly faster.
I've spent an awful lot of time tinkering about with this and I regret not being slightly more disciplined and scientific in my approach now.
Provisionally it seems the safest thing to do until we can work out whether dry ferts are a better option is just to reduce the amount of fishmix by half. I don't believe it'll have a detrimental effect on how fast the algae grows, maybe only the ultimate cell density of the cultures.

Lol I didn't realise you'd just randomised the quantity of fish mix. Given that, I will try halving the next lot of fish mix I use and do 1/2ml per litre. In that study I linked to back in this thread, where they were testing inorganic ferts, they did find that a certain level of ferts did produce the optimum level of Chlorella - so having too much ferts did reduce production in the same way as having too little.

I've not actually had chance to test a drop of fish mix in tap water with the ammonia test kit to double check that it does actually contain ammonia, but it seems likely as you suggested. I just can't understand, if that is the case, why you or Miranda don't have any issues with ammonia, and it is an issue isolated to me. I also can't quite understand why ammonia is an issue at all if commercially moina and daphnia are cultured over a bed of animal or chicken manure - surely the ammonia levels in water full of manure would be off the chart?

The dreaded yellow/green clouding is coming back again in my culture, so I'm starting to wonder if I need to do a full reset, and drain and clean the tank. It is getting a little frustrating.

I have three empty 2 litre bottles now, so I'm going to set them up as test cultures on the window sill. One with 1ml per litre fish mix, one with 1/2 ml per litre fish mix, and one with dry salt ferts. I'll probably set them up with a reflector behind them to to maximise the light input. Although I won't be able to run an air stone through them, I'll shake them twice a day, and see what production is like in each.
 

louis_last

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The dreaded yellow/green clouding is coming back again in my culture, so I'm starting to wonder if I need to do a full reset, and drain and clean the tank. It is getting a little frustrating.

I have three empty 2 litre bottles now, so I'm going to set them up as test cultures on the window sill. One with 1ml per litre fish mix, one with 1/2 ml per litre fish mix, and one with dry salt ferts. I'll probably set them up with a reflector behind them to to maximise the light input. Although I won't be able to run an air stone through them, I'll shake them twice a day, and see what production is like in each.
It's weird but this basically sounds like when people get cloudy water in their aquariums doesn't it? I've often seen threads where people can't work out what caused it and it keeps coming back after water changes only to eventually disappear as quickly as it started. The weird thing is that you can actually use daphnia and moina to clear algal and bacterial blooms in aquariums - I literally did recently but taking my filter out and overdosing moina for a few days and they cleared it right up.
I've had various things explode in my cultures, rotifers, and at the moment there's a lot of paramecium in my main culture, but never a free floating bacteria that the moina don't eat - yet.
It might be interesting to take two cupfulls of your culture water with some moina and the unusual yellow clouding and continue to feed one chlorella and the other yeast for a few days to see what happens.
I keep thinking about how the autotrophic cultures of chlorella are yellow too. I fed some chlorella that had definitely used up all the fishmix to a pint glass culture with clear water last night and today this morning they've cleared it and there's no sign of brown hint or anything else untoward in the water.
Do you have any idea what your tap water parameters are like? Mine is soft to the extent that when I run it through a distiller there's almost no minerals left behind. At my old house it literally left behind a thick coating of chalky, rusty gunk every time even from just 4L of water.
 

Wookii

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It's weird but this basically sounds like when people get cloudy water in their aquariums doesn't it? I've often seen threads where people can't work out what caused it and it keeps coming back after water changes only to eventually disappear as quickly as it started. The weird thing is that you can actually use daphnia and moina to clear algal and bacterial blooms in aquariums - I literally did recently but taking my filter out and overdosing moina for a few days and they cleared it right up.
I've had various things explode in my cultures, rotifers, and at the moment there's a lot of paramecium in my main culture, but never a free floating bacteria that the moina don't eat - yet.
It might be interesting to take two cupfulls of your culture water with some moina and the unusual yellow clouding and continue to feed one chlorella and the other yeast for a few days to see what happens.
I keep thinking about how the autotrophic cultures of chlorella are yellow too. I fed some chlorella that had definitely used up all the fishmix to a pint glass culture with clear water last night and today this morning they've cleared it and there's no sign of brown hint or anything else untoward in the water.
Do you have any idea what your tap water parameters are like? Mine is soft to the extent that when I run it through a distiller there's almost no minerals left behind. At my old house it literally left behind a thick coating of chalky, rusty gunk every time even from just 4L of water.

Yes, it does seem very much like a cloudy bacterial bloom some people see on a new tank, however it is not just white clouding as usually seen, it definitely has green/yellow colouration to it. It definitely grows and accumulates over time pretty rapidly. As you say, if it was a straight forward bacterial bloom, you would have thought such a high density of Moina would have cleared it up in no time.

It is quite possible that there is more than one thing going on though. There could be a growth of Chlorella in the tank at the same time as the bacterial bloom, with the high lights, and the fairly nutrient rich tap water. Though again, I don't know how it can cloud so quickly if so many Moina are actively feeding.

My tap water is fairly hard - KH 6 and GH 12 or there abouts, usually a fair amount of Magnesium in there too, not just Calcium. Some nitrates per the water report, but nothing untoward - I use it in my low tech tank without issues.

Not that I expect you to go out and buy a test kit, but it would be useful to know if your fully matured Chlorella culture still has any ammonia in. I'm assuming not, as I guess your snails would soon tell you by all sitting up above the waterline as mine did.

I haven't checked the pH of the Moina tank - given the KH of the tap water, I didn't think it worthwhile as it should be well above 7.
 

louis_last

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Not that I expect you to go out and buy a test kit,
I'm going to don't worry.
but it would be useful to know if your fully matured Chlorella culture still has any ammonia in. I'm assuming not, as I guess your snails would soon tell you by all sitting up above the waterline as mine did.
I can't say I've ever seen the snails do that, sometimes they have clustered near the surface right after a feed but that happened even with yeast.
I haven't checked the pH of the Moina tank - given the KH of the tap water, I didn't think it worthwhile as it should be well above 7.
Soon I'll be able to let you know the ph of my culture and what results im getting ammonia wise.
 

Wookii

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So I tested the Fish Mix last night - a tiny dip of a pipette tip, mixed in some tap water - for ammonia, and sure enough the colour went off the chart, so that's my ammonia source as you suspected Louis.

I have decanted my current main Chlorella culture into 2 litre plastic bottles, as even though its very dark green, its still reading ammonia. They will stay on the window sill until I no longer measure any ammonia. In the empty culture container (in which I left in about half a pint of the old culture), I've added fresh tap water with some prime and a dose of macros and micros from my high tech. The 4 litre tub has basically had what would be an EI dose for 100 litres. It does contain humic/fluvic acids etc which I add to my micros and macros, so hopefully this won't be an issue for the Chlorella - I'm assuming not as long as the pH stays above 7. I can always add some more potassium carbonate if need be. When I have more time I'll mix up a dedicated mix for the cultures.

I've also moved the light, and instead of it sitting on the top of the tall narrow culture tubs, it sits at the back shining through the middle of them, and seems to illuminate the inside of the tubs much better, so hopefully that will help going forward.
 

louis_last

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It's entirely possible that the ratio of nutrients in the fish mix is such that all of some necessary nutrient for the chlorella is used up before all of the ammonia is purged that's why I'm keen to be able to test mine but it wouldn't necessarily explain why me and miranda don't seem to be having the same issue. It would be another huge point in favour of dry salts though. Lacking any dry salts I'm going to try a bottle with tropica specialised fertiliser and see how that goes.
The main reason I initially went with fishmix was that I'd used it before and it had the sugar in it. The guy from the parosphromenus project who really seemed to know what he was talking about when it came to chlorella and moina, and was speaking from experience, was also adamant that a 6-1 N-P ratio was ideal. Fishmix is 5-1 and was the closest I could find at the time.
Another thing to consider is that being an organic product the fish mix is likely to vary slightly from bottle to bottle.
I'm not at all married to the idea of using fishmix, I'd be really happy to find something that works better.
 

louis_last

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Got my microfex from the Russian guy Alex today so I've got another variable for the moina cultures to keep things nice and confusing now. He also confirmed that in addition to maintaining several specific cultures of them (that he apparently really does harvest 500g from a day) he also keeps them on the bottom of his moina cultures. He actually mentioned this without me asking as a good way to keep a back up culture in case the main ones of worms crash.
 

louis_last

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Is that the guy on ebay @louis_last?
If so I just bought some off him too lol
There's two guys on ebay, one of them is shipping the worms from Denmark and the other is Alex the Russian guy I'm talking about.
I was a test case to see how they would survive shipping and they seem to have survived just fine. If it's Alex you've ordered them from you wont be disappointed with what he sends although the cultures do smell worse than I imagined.
He's a very helpful guy with a lot of knowledge, I've spoken to him on the phone a couple of times and he'll be very happy to help you with anything to do with the worms. I've not tried feeding any yet but I'm sure the fish will love them.
He didn't charge me for mine because it was a test so it seems like the least I can do is leave a LINK here to his listing for anyone who might want a great starter culture of these worms now that he's able to ship them. He packs them for transit really well and he's located in the UK so they're not stuck in transit for a long time, apprently they are very sensitive to being moved and don't handle shipping well at all.
 
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