- Joined
- May 22, 2016
- Messages
- 6,970
- Reaction score
- 10,747
[Edits for clarity - 4/21 - shown in brackets]
I've been looking into debris that accumulates in the sand (Dan's Thread for that discussion) in our tanks and ways to "clean it up" whatever that means. In the process Waste Away by Dr Tim and similar "grunge-eating" products became of interest.
Although it "works", it does not seem to behave as many would expect - like a bacterial additive - and so the way we use it ought to reflect that.
I'll present two hypotheses of what Waste Away is and how it functions, then lay out the experiments done that might shed light on the question.
Hypothesis 1: "Waste Away is a bacterial additive that contains 8 strains/species [Dr Tim] that consume uneaten food and waste in a system. Adding it is effective because the system lacks significant numbers of these bacteria."
Hypothesis 2: "Waste Away is a growth medium for bacteria that is rich in Carbon and contains biologically significant amounts of Phosphorus. Adding it is effective because it allows existing bacteria in a system to grow and multiply, in the process consuming Nitrogen sources where the bacteria multiply."
I'll just do barebones experiments and headlines to try to condense this somewhat.
Experiment 1:
10mL test tubes of aquarium grunge had various bacterial products added to them consistently over a month with or without carbon dose.
Result: neither Waste Away or any other product or combination caused this material to decrease in any measurable way. Some of the tubes - WA and all tubes with carbon added - clouded and went anaerobic.
Maybe they needed aerobic digestion conditions to consume the grunge (nope).
[edit: 4/21 - the most likely interpretation of experiment 1 & 2 is that this material is mostly indigestible]
Experiment 2: A tube of grunge had air slow bubbled through it, WA and over 60+ days every combination of C, N, and P were rotated through to see if any aided reduction of material. Material amount and various water properties were measured.
Neither W.A. alone or supplemented with any nutrient (C,N,P) or any combination caused the material to decrease in any measurable way.
WA addition caused NO3 in water to disappear, and PO4 went up. Maybe this was due to a breakdown of PO4-containing material in the grunge (nope).
[edit: 4/21 - the most likely interpretation of experiment 1 & 2 is that this material is mostly indigestible]
Experiment 3: Direct N & P tests of what comes out of the Waste Away Bottle.
4ml/L of Waste Away (16x the recommended dose concentration)
PO4 = 0.5ppm PO4 (implies a +0.03ppm PO4 dose to a system when using recommended dose)
NO3 = undetectable
Ammonia = undetectable
these results were consistent on 2 different bottles and whether the test was done on cloudy solution straight from the bottle, or when the contents were filtered through a 0.22micron syringe filter to remove bacteria first. The media is a small phosphate dose.
Experiment 4: measure the oxygen consumed (and therefore organic carbon oxidized) in a bottled aquarium sample when Waste Away is added.
If WA bacteria were consuming organics in the aquarium sample, then repeated doses of WA would yield smaller and smaller O2 consumption as it "cleaned" the sample of organics, but that doesn't happen. If on the other hand, the bacteria in the aquarium sample were consuming organic carbon in the Waste Away media, then it would look exactly like the above data - every additional dose of WA starts a new round of rapid O2 consumption.
Experiment 5: Separate Waste Away by centrifuging, see which causes the oxygen consumption, the bacterial pellet at the bottom of the centrifuge tube, or the sterilized media.
The bacterial pellet from WA seemingly did nothing. The media with the bacteria removed, and sterilized by boiling was responsible for all the consumption of organics that is seen in whole Waste Away. (Difference between whole [blue] and sterilized [yellow] is that boiling the WA media slightly increased its concentration above that of the raw whole WA.)
Experiment 6: direct microscope observation of WA bacteria.
There they are! wiggling and alive? (nope)
except then I boiled, froze, and bleached the sample. And under the microscope they looked exactly the same! This motion is simply the random motion of inert particles.
Experiment 7: Direct chemical analysis of WA from @Dan_P
PO4: at recommended dose of 1mL /Gal adds +0.032ppm (126ppm in bottle itself)
NO3: undetected
NH3: undetected
Amines / Other Nitrogen species: Low (insignificant compared to PO4 numbers)
Organic Carbon: very high - a tentative ~5000ppm Carbon
This tracks my results using a a different meter stick, based on 3 day oxygen consumption from my multipole rounds of tests, Waste Away seems to have in the ballpark of 1/6th of the organic carbon of pure vodka.
Finally experiment 8: Can Waste Away bacteria be cultured?
Created a bacteria-freindly medium of new salt water + f/2 + tiny pinch of fish flake + Carbon dose. Split the media in 10 containers - four 60mL flasks and six 10mL test tubes. Boiled all to sterilize and stoppered them while hot. After cooling to room temp I added drops of WA to the containers. Half of them got WA that had been boiled to sterilize and the other half got raw WA out of the bottle. Cultured in the dark.
After 5 days there was no difference in the bactierial growth/cloudiness between the sterilized WA and the raw WA. Then to prove the media would happily grow live bacteria, on day 5 I added a drop of aquarium water to one container of each treatment kind - 4 total containers.
As you can see, there was no difference in the bacterial growth/cloudiness between sterile, boiled Waste Away (Blue) and Raw Waste away (Red). The dashed lines that showed dramatic cloudiness and bacterial growth were the 4 samples that got drops of aquarium water on day 5.
Summary:If there's viable effective bacteria in Waste Away that do anything important in an aquarium context, I have no idea how to prove it. All the tests that I can come up with show the opposite.
[Edit: 4/21 - These tests showed some large effects from Waste Away in the context of aquarium water, but the bacteria in WA were not responsible for any of these easily measurable effects.]
On the other hand, the effects of Waste Away are easily demonstrated, but a close analysis shows that all those effects can be accounted for by the chemistry of the Waste Away media.
(I have a hunch the same is true for some other bacteria products)
I like and will continue to use Waste Away, but as a carbon source / nitrogen reduction / bacterial growth medium, not as a source for bacteria themselves.
Did I miss anything, @Dan_P ?
Edit 4/18: Dr. Tim response is here (post 80)
Edit 4/18 pm: My response to Dr. Tim's criticisms is here (post 90)
I've been looking into debris that accumulates in the sand (Dan's Thread for that discussion) in our tanks and ways to "clean it up" whatever that means. In the process Waste Away by Dr Tim and similar "grunge-eating" products became of interest.
Although it "works", it does not seem to behave as many would expect - like a bacterial additive - and so the way we use it ought to reflect that.
I'll present two hypotheses of what Waste Away is and how it functions, then lay out the experiments done that might shed light on the question.
Hypothesis 1: "Waste Away is a bacterial additive that contains 8 strains/species [Dr Tim] that consume uneaten food and waste in a system. Adding it is effective because the system lacks significant numbers of these bacteria."
Hypothesis 2: "Waste Away is a growth medium for bacteria that is rich in Carbon and contains biologically significant amounts of Phosphorus. Adding it is effective because it allows existing bacteria in a system to grow and multiply, in the process consuming Nitrogen sources where the bacteria multiply."
I'll just do barebones experiments and headlines to try to condense this somewhat.
Experiment 1:
10mL test tubes of aquarium grunge had various bacterial products added to them consistently over a month with or without carbon dose.
Result: neither Waste Away or any other product or combination caused this material to decrease in any measurable way. Some of the tubes - WA and all tubes with carbon added - clouded and went anaerobic.
Maybe they needed aerobic digestion conditions to consume the grunge (nope).
[edit: 4/21 - the most likely interpretation of experiment 1 & 2 is that this material is mostly indigestible]
Experiment 2: A tube of grunge had air slow bubbled through it, WA and over 60+ days every combination of C, N, and P were rotated through to see if any aided reduction of material. Material amount and various water properties were measured.
Neither W.A. alone or supplemented with any nutrient (C,N,P) or any combination caused the material to decrease in any measurable way.
WA addition caused NO3 in water to disappear, and PO4 went up. Maybe this was due to a breakdown of PO4-containing material in the grunge (nope).
[edit: 4/21 - the most likely interpretation of experiment 1 & 2 is that this material is mostly indigestible]
Experiment 3: Direct N & P tests of what comes out of the Waste Away Bottle.
4ml/L of Waste Away (16x the recommended dose concentration)
PO4 = 0.5ppm PO4 (implies a +0.03ppm PO4 dose to a system when using recommended dose)
NO3 = undetectable
Ammonia = undetectable
these results were consistent on 2 different bottles and whether the test was done on cloudy solution straight from the bottle, or when the contents were filtered through a 0.22micron syringe filter to remove bacteria first. The media is a small phosphate dose.
Experiment 4: measure the oxygen consumed (and therefore organic carbon oxidized) in a bottled aquarium sample when Waste Away is added.
If WA bacteria were consuming organics in the aquarium sample, then repeated doses of WA would yield smaller and smaller O2 consumption as it "cleaned" the sample of organics, but that doesn't happen. If on the other hand, the bacteria in the aquarium sample were consuming organic carbon in the Waste Away media, then it would look exactly like the above data - every additional dose of WA starts a new round of rapid O2 consumption.
Experiment 5: Separate Waste Away by centrifuging, see which causes the oxygen consumption, the bacterial pellet at the bottom of the centrifuge tube, or the sterilized media.
The bacterial pellet from WA seemingly did nothing. The media with the bacteria removed, and sterilized by boiling was responsible for all the consumption of organics that is seen in whole Waste Away. (Difference between whole [blue] and sterilized [yellow] is that boiling the WA media slightly increased its concentration above that of the raw whole WA.)
Experiment 6: direct microscope observation of WA bacteria.
There they are! wiggling and alive? (nope)
except then I boiled, froze, and bleached the sample. And under the microscope they looked exactly the same! This motion is simply the random motion of inert particles.
Experiment 7: Direct chemical analysis of WA from @Dan_P
PO4: at recommended dose of 1mL /Gal adds +0.032ppm (126ppm in bottle itself)
NO3: undetected
NH3: undetected
Amines / Other Nitrogen species: Low (insignificant compared to PO4 numbers)
Organic Carbon: very high - a tentative ~5000ppm Carbon
This tracks my results using a a different meter stick, based on 3 day oxygen consumption from my multipole rounds of tests, Waste Away seems to have in the ballpark of 1/6th of the organic carbon of pure vodka.
Finally experiment 8: Can Waste Away bacteria be cultured?
Created a bacteria-freindly medium of new salt water + f/2 + tiny pinch of fish flake + Carbon dose. Split the media in 10 containers - four 60mL flasks and six 10mL test tubes. Boiled all to sterilize and stoppered them while hot. After cooling to room temp I added drops of WA to the containers. Half of them got WA that had been boiled to sterilize and the other half got raw WA out of the bottle. Cultured in the dark.
After 5 days there was no difference in the bactierial growth/cloudiness between the sterilized WA and the raw WA. Then to prove the media would happily grow live bacteria, on day 5 I added a drop of aquarium water to one container of each treatment kind - 4 total containers.
As you can see, there was no difference in the bacterial growth/cloudiness between sterile, boiled Waste Away (Blue) and Raw Waste away (Red). The dashed lines that showed dramatic cloudiness and bacterial growth were the 4 samples that got drops of aquarium water on day 5.
Summary:
[Edit: 4/21 - These tests showed some large effects from Waste Away in the context of aquarium water, but the bacteria in WA were not responsible for any of these easily measurable effects.]
On the other hand, the effects of Waste Away are easily demonstrated, but a close analysis shows that all those effects can be accounted for by the chemistry of the Waste Away media.
(I have a hunch the same is true for some other bacteria products)
I like and will continue to use Waste Away, but as a carbon source / nitrogen reduction / bacterial growth medium, not as a source for bacteria themselves.
Did I miss anything, @Dan_P ?
Edit 4/18: Dr. Tim response is here (post 80)
Edit 4/18 pm: My response to Dr. Tim's criticisms is here (post 90)
Last edited: