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If his ppm is 30, it matters little whether it is GH or KH. The water is soft. Chestnut bark is acidic, so I think that's the reason.
Did you make sure that the chestnut stump has been dead for some time (completely dry)? You want to ensure there are no sugars from a recently living plant...
Sure-- I wasn't making a claim about whether or not there would be aggression with this species or all species. Just that adding an additional female introduces variables that are often not taken into account when the general advice is given to keep trios to avoid too much attention/stress on...
I've kept trios of apistos (not agassizii) in a 20-long that was planted and scaped such that you could not see the other side of the aquarium either from the front or from either side. In my experience, having a third fish present makes the social dynamic more complicated (not necessarily...
I’ve used air hose before to keep plants separated from the air pump output. But I always had protein scum accumulate on the still portion, so I eventually stopped doing it.
Here are things I've used my 2.5 gallon for:
Breeding pairs of small fish, like many killifish or parosphromenus. They go back to the larger tank after spawning, however.
Medication, recovery, or separation of small fish (less than 2" in size).
Inverts (asellus, shrimp, or blackworms)
It's possible if you're using leaded crystal (quite unlikely) or painted glass (possible). But the amount of lead that would leach into the water would only be a few ppm, and it might take weeks to get to that point.
I don't think I could conclude anything from the results you've given, but I also don't think the substrate was leaching (independent of your test). If there's a diffrence with fresh substrate, I would think it's from debris that will eventually be rinsed away.
In the future, it’s good to do all the treatments at the same time so the only thing that’s different is the thing you’re varying. i.e., the water starts the exact same, temp is always the same, etc.