Waste Away: Is it really bacterial? Or chemical? What does it do?

[taricha]

@taricha Any updates? I'm really curious.

I should have updates on my nutrient reduction bottles tomorrow!

Creating aquarium-ish conditions that culture up WA bacteria from a bottle is more challenging to demonstrate.

 

[taricha]

Follow-up on this nutrient reduction exercise...

Here's a quick demonstration that anyone on this chem forum could replicate.
Four identical 500mL bottles get tank water enriched* with NO3 and PO4, I went to around 50ppm NO3 and 1.5ppm PO4 so roughly 10x my typical tank levels.

Bottles equally aerated in the dark at room temp. After a couple of days, make sure nutrients are still tracking similarly in the 4 bottles, then add the different treatments.
1 gets nothing, 2 gets recommended dose (1/8mL) of normal Waste-Away, 3 gets sterilized WA (frozen/boiled), 4** gets same dose of WA bacteria/spores - no WA media.

every 2 to 3 days Check NO3, PO4 and add another recommended dose of each treatment.

Results? Will the NO3 and PO4 consumers show effects?
We'll see in a week!

*Micro-Algae Grow from Florida aqua farms Guillard "f/2" medium + ascorbic acid
**(the only part most people can't do) centrifuged 5 min 4000rpm, supernatant poured off, pellet of solids/bacteria/spores resuspended in equal volume tank water.

here's some data on this.

This set got the treatments added at Day2. The nutrient reductions are small (just one dose and just 2 days), but the recommended dose of WA shows same nutrient reduction profile as the sterilized media. 10% error bars show that NO3 may not be significant, but PO4 probably is. This test had to be stopped early because my 10 month old managed to topple the control bottle. So I literally have no control for that one. oops.
And thus here's the re-do...

Two treatment doses at day 1.5 and 3.5. The WA and the sterilized media of WA show the same large PO4 decrease. The separated pellet of WA spores shows no nutrient reduction compared to the Control, difference could be measurement error.
Interestingly, the NO3 doesn't seem to budge much. This is out of sync with my NO3 reduction results from post 47 [edit: although in post 47, bottles got double dose, there were more treatment additions, and experiment went longer so differences more dramatic than the current]

So What DOES Waste Away Do?
....

So here's a demo of what WA (a carbon-rich media) does in terms of nutrient reduction and comparing that with a straight carbon source, vodka.
...

Stars indicate which days WA and Vodka were added....

The difference [other than amount and duration of dosing] between these two tests was that in the one that showed NO3 drop, I "skimmed" the bottles with an airstone and paper towels at the bottle opening. I didn't think It was important, so I omitted it from the current run, initially.

To check and see if skimming the bottles can bring the two sets of NO3 results back in closer agreement, I started skimming the bottles at day 5.... and voila.

the treatment that got a carbon dose via WA and the one that got the boiled/frozen WA media both showed faster NO3 drops when skimming started at day 5. Additionally, the water cloudiness clearly shows the effect of the skimming at removing the cloudy bacterial bloom that the WA and the WA-media caused.

Summary: Overall, the results are pretty much as expected, the addition of carbon dose from WA and the boiled/frozen media of WA each caused a bacterial bloom that clouded the water, and reduced PO4. Unexpectedly, skimming was required for NO3 decrease to show up.
The treatment that contained only a recommended dose of the WA bacteria/spores (with the carbon-rich media removed) showed no detectable nutrient reduction relative to the control. This may be because the carbon sources in the water sample were not easily available (in form or amount) to the WA bacterial spores.

 

[Michael Gray]

I think I'm like day 10 of dosing wa for Dino's. Now that I'm dosing 80ml. My po4 tanks roughly 4 hours. From 47ppb to about 6ppb dose at 11am skimmer/uv off. And 3pm test 6ppb. Quickly dose to raise to .1ppm again

 

[RMS18]

Follow-up on this nutrient reduction exercise...


here's some data on this.

This set got the treatments added at Day2. The nutrient reductions are small (just one dose and just 2 days), but the recommended dose of WA shows same nutrient reduction profile as the sterilized media. 10% error bars show that NO3 may not be significant, but PO4 probably is. This test had to be stopped early because my 10 month old managed to topple the control bottle. So I literally have no control for that one. oops.
And thus here's the re-do...

Two treatment doses at day 1.5 and 3.5. The WA and the sterilized media of WA show the same large PO4 decrease. The separated pellet of WA spores shows no nutrient reduction compared to the Control, difference could be measurement error.
Interestingly, the NO3 doesn't seem to budge much. This is out of sync with my NO3 reduction results from post 47


The difference between these two tests was that in the one that showed NO3 drop, I "skimmed" the bottles with an airstone and paper towels at the bottle opening. I didn't think It was important, so I omitted it from the current run, initially.

To check and see if skimming the bottles can bring the two sets of NO3 results back in closer agreement, I started skimming the bottles at day 5.... and voila.

the treatment that got a carbon dose via WA and the one that got the boiled/frozen WA media both showed faster NO3 drops when skimming started at day 5. Additionally, the water cloudiness clearly shows the effect of the skimming at removing the cloudy bacterial bloom that the WA and the WA-media caused.

Summary: Overall, the results are pretty much as expected, the addition of carbon dose from WA and the boiled/frozen media of WA each caused a bacterial bloom that clouded the water, and reduced PO4. Unexpectedly, skimming was required for NO3 decrease to show up.
The treatment that contained only a recommended dose of the WA bacteria/spores (with the carbon-rich media removed) showed no detectable nutrient reduction relative to the control. This may be because the carbon sources in the water sample were not easily available (in form or amount) to the WA bacterial spores.

The bacterial Bloom you refer to in the summary, where did that bacteria that bloomed originate from? WA or the tank water?

 

[taricha]

The bacterial Bloom you refer to in the summary, where did that bacteria that bloomed originate from? WA or the tank water?

Good Q.
The bacterial bloom appeared to happen equally in the treatment that got actual Waste Away and the one that got the WA media only, having been centrifuged, boiled, & frozen. So seems like the cloudiness in that case was caused by the media triggering aquarium bacteria to bloom.

This test at least didn't show signs of WA bacteria blooming or reducing nutrients.

Currently messing around with conditions that ought to cause the WA bacteria to show some effects. We'll see...

 

[Grigs]

I don't really want to subject everyone to re-read my original post which is #80. Did you get a chance to read it?

A quick summary:
Waste-away was used a too high a concentration - more is not better.
There is no evidence that the 'grunge' used in the test was biodegradable - no controls.

How Waste-Away works -
Waste-Away is a combination of several heterotrophic bacteria that I isolated from aquariums and I found are good at consuming nitrate and phosphate and degrading organics. I do not buy bacteria from the ACME Bacteria Company - I isolated these.

Organic material is constantly added and/or produced in an aquarium. It can be dead organisms, feces, dead algae and bacteria, fish food, etc. Bacteria degrade or mineralize this material. If nature did not have this process nothing would decay. But some bacteria will produce exopolymer susbtances (EPS) etc and form a biofilm - that stick coating or slime on the tubing, pipes, filter pads etc. clogging these. Other bacteria (like some in Waste-Away) can be used to degrade this EPS and reducing clogging. But as I mentioned in the another post - not 100% of the material can be degraded that is why you get a build of material over time in the substrate and other areas.

As Sanjay and others showed years ago in an article in Advanced Aquarist on-line - your skimmer removes a significant percentage of bacteria from the water and it seems to be selective. As you reduce the bacteria in the water column you give bacteria/other organism that live on surfaces a better chance to survive because your have reduce their competitors. This can lead to cyano and GHA etc because they are not removed by the skimmer.

The general idea behind bacteria additives is to add back the bacteria the skimmer has removed. Now that assumes there is enough carbon in water which maybe is not always the case.

A product like RedSea NOPOX (or vodka dosing) assumes that there is not enough carbon in the water and that by adding carbon you will promote the bacteria already in the water to grow and assimulate the NO3 and PO4.

The conclusion of the OP was that my claim that Waste-Away worked by adding bacteria to the water which remove NO3 and PO4 was wrong based on his tests and that Waste-Away works solely because it acts as a carbon source like vodka or NOPOX.

Regards

This post was the response that I had hoped to read from you. I REALLY appreciate you taking the time to write it. Thank you.

 

[SMSREEF]

@taricha What do you think about doing this test with new saltwater rather than tank water?

the reason I’m thinking this may show results is because I recently am getting over Dino’s and wanted to do a large water change. However, I wanted to make sure I replaced the old water with new water loaded with bacteria to hopefully keep Dino’s under control as I pulled my UV sterilizer from the tank.

I made the water and heated to 82, dosed Dr Tim’s WA, Microbacter 7, ZEObac, and phosphate, nitrate and some vinegar for a carbon source. I cooked it for 2 days and looked under a microscope. I did see microbes (I am not a microbiologist so I have no clue what kind I saw). This was in just a drop of the 5 gallon bucket.

this leads me to think you may be able to take out the confounding factor of tank bacteria by using newly mixed saltwater. You could use a carbon source as a variable.

Another thought, if the bacillus bacteria Dr Tim has in Waste away is a spore former, it possibly can live through the freezer and less likely but possibly boiling...

Boiling and Bacillus Spores

www.ncbi.nlm.nih.gov

 

[RMS18]

Good Q.
The bacterial bloom appeared to happen equally in the treatment that got actual Waste Away and the one that got the WA media only, having been centrifuged, boiled, & frozen. So seems like the cloudiness in that case was caused by the media triggering aquarium bacteria to bloom.

This test at least didn't show signs of WA bacteria blooming or reducing nutrients.

Currently messing around with conditions that ought to cause the WA bacteria to show some effects. We'll see...

Understood... At this point in testing and future tests may show different results, WA is a $$ carbon source.?

 

[biom]

this leads me to think you may be able to take out the confounding factor of tank bacteria by using newly mixed saltwater. You could use a carbon source as a variable.

the newly mixed saltwater isn't sterile, one of the reasons is the use of natural sodium chloride in mixtures, which contains lots of bacteria. Actually they say this is the reason why Red Sea's salts are so successful, because they are using natural salt from Red Sea.

Another thought, if the bacillus bacteria Dr Tim has in Waste away is a spore former, it possibly can live through the freezer and less likely but possibly boiling...

Correct, if I remember correctly sterilization should meet some standard parameters - autoclaving in 120 degrees Celsius for 15 minutes or so. Not sure how OP sterilized media, but this is not scientific paper and we should not expect "materials and methods" section .

 

[SMSREEF]

the newly mixed saltwater isn't sterile, one of the reasons is the use of natural sodium chloride in mixtures, which contains lots of bacteria. Actually they say this is the reason why Red Sea's salts are so successful, because they are using natural salt from Red Sea.


Correct, if I remember correctly sterilization should meet some standard parameters - autoclaving in 120 degrees Celsius for 15 minutes or so. Not sure how OP sterilized media, but this is not scientific paper and we should not expect "materials and methods" section .

Oh!!! I do use Red Sea salt. There goes that idea...
But even more bacteria for my tank so at least that’s good.

 

[taricha]

@taricha What do you think about doing this test with new saltwater rather than tank water?
...
this leads me to think you may be able to take out the confounding factor of tank bacteria by using newly mixed saltwater.

If we're trying to say something about what happens when WA is added to an aquarium, then aquarium bacteria are not a confounding variable, but a necessary condition to that sort of question.
On the other hand, when I'm focusing on simply trying to demonstrate that WA is being cultured, I don't make them compete with aquarium microbes.

I made the water and heated to 82, dosed Dr Tim’s WA, Microbacter 7, ZEObac, and phosphate, nitrate and some vinegar for a carbon source. I cooked it for 2 days and looked under a microscope. I did see microbes (I am not a microbiologist so I have no clue what kind I saw). This was in just a drop of the 5 gallon bucket.

Jut for curiosity sake (I'm curious) do two 5 gal buckets side by side - spike each with N, P, and vinegar, add the recommended dose of Bac bottles to one, and not the other. To replace the carbon content in WA, add an extra ~1/6mL of vodka (or 1.3mL vinegar) per mL of WA. That's my best estimate of the carbon content of WA. Since MB7 heavy doses aren't known (as far as I am aware) to cause cloudy water events, then it's likely not a significant carbon dose. Zeo, I have no guidance on. Maybe leave it out.
See what grows and clouds up in two days.
Biom suggests here how it might go.

Actually they say this is the reason why Red Sea's salts are so successful, because they are using natural salt from Red Sea.

But even more bacteria for my tank so at least that’s good.

If you were really curious, you could add a 3rd bucket - Instant Ocean and see if the clearly evident bacterial bloom is observably different between the salt mixes and the new SW with bac bottles.
(my guess is they might all go cloudy).

Another thought, if the bacillus bacteria Dr Tim has in Waste away is a spore former, it possibly can live through the freezer and less likely but possibly boiling...

Boiling and Bacillus Spores

www.ncbi.nlm.nih.gov

they are described as spores of bacillus.

Correct, if I remember correctly sterilization should meet some standard parameters - autoclaving in 120 degrees Celsius for 15 minutes or so. Not sure how OP sterilized media,

yeah, pasteurization is more accurate. I centrifuged them to concentrate the spores in a visible pellet, poured off the supernatant "media" boiled/froze the media, then boiled again just before addition.
I certainly didn't get them all, but it should have created dramatically different amounts of bacteria between the boiled/frozen media (clear) and the raw WA (quite cloudy).

 

[taricha]

Understood... At this point in testing and future tests may show different results, WA is a $$ carbon source.?

The bacteria in WA are likely viable, culture-able and perhaps even important in aquaria, but in tests so far, whatever effect they have is obscured by the fireworks of a carbon dose, and the response of native aquarium bacteria. Future tests will try harder to look past the fireworks.

 

[biom]

If you were really curious, you could add a 3rd bucket - Instant Ocean and see if the clearly evident bacterial bloom is observably different between the salt mixes and the new SW with bac bottles.

Here is a very good reading about bacteria in reef, reef tanks and salts

Bacterial Counts in Reef Aquarium Water: Baseline Values and Modulation by Carbon Dosing, Protein Skimming, and Granular Activated Carbon Filtration

What are the bacteria populations in the water column of reef tanks, and how does that value compare with bacterial counts in authentic reef water? Does carbon dosing indeed increase water column bacteria populations (i.e., is growth carbon limited)? Does mechanical filtration (protein skimming...

reefs.com

from this article: bacteria count of different brands of salts

Bacteria/mL count of six different brands of salt mix made up to 35 ppt salinity using RO/DI/0.2 uM filtered water in capped and foil-wrapped sterile flasks

 

[biom]

Thank you @taricha for your curiosity and all experiments you are doing. The results are really as expected but the graphs are bit confusing because of the different colors you have used for same parameters for example WA is once red and then green, media is yellow , then red, etc. And at the Cloudiness graph is probably mistake, because the pellet gives greatest bacterial cloudiness.

Interestingly, the NO3 doesn't seem to budge much. This is out of sync with my NO3 reduction results from post 47

in experiment from post #47 you were using double recommended dose of Waste away.

 

[taricha]

from this article: bacteria count of different brands of salts

I forgot how much ground is covered in the Feldman article - that they did salt mixes too.

The results are really as expected but the graphs are bit confusing because of the different colors you have used for same parameters ...
And at the Cloudiness graph is probably mistake, because the pellet gives greatest bacterial cloudiness.

Thanks. That was messy. Edited the post and cleaned up the graphs.

in experiment from post #47 you were using double recommended dose of Waste away.

and did more doses and longer duration. so a few reasons in addition to skimming that NO3 decrease showed up better in the earlier exercise.

 

[RevMH]

Thank you for this. I'm finding it extremely interesting and informative.

 

[wackyreefer]

Thank you for this. I found this very informative and well timed for me since I am dealing with Dino's and am busy dosing WA.

I myself have noticed that after the first dose (half dose) of WA, it has plummeted my phosphate to zero within hours (Which I currently dose daily to maintain a reading of 0.03-0.1) and ever so slightly reduced nitrates, but not completely (which matches WA product marketing and @taricha 's findings) which is great.

I do have a question from a hobbyist point of view:

Considering WA is labeled to "Lowers phosphates & Nitrates ", " Combats Slime" and " Dissolves sludge and dirt " , and many stores market it on their websites as a goto for Dinoflagellates removal. (now I am unsure if DrTims is marketing it as a Dinoflagellates solution or if the stores selling it are adding this, regardless this is merely for context).

If your Dinoflagellates are as a result of zero nutrients (nitrate and phosphate), and according to Dr Tim WA reduces nutrients (nitrate and phosphate - which @taricha 's test has also shown), then my simpleton brain starts wondering if it's the right product for you to combat Dinoflagellates?
Now I am happy to be educated / corrected here, as I want to ensure I understand this correctly.

Would WA not continuously keep lowering your nutrients creating the same environment that helped establish the Dinoflagellates?
Or is the consumption of nutrients 'activating spores of bacteria', developing a community of bacteria that eventually over time will out compete the Dinoflagellates for available nutrients forcing the Dinoflagellates to slowly die off?

 

[Zerobytes]

I can only speak on results but I dose WA every 3-4 months (2 1/2 dosages over 3 or 4 days). I get a reduction in NO3 and PO4, but nothing drastic. I don’t dose anything else but Alk and Cal. Doesn’t seem to cause any issues but the tank and sand appear cleaner. I do turn off the skimmer for 8 hours or so. It goes a bit nuts when turned back on, even with a water change.

 

[salty150]

I can only speak on results but I dose WA every 3-4 months (2 1/2 dosages over 3 or 4 days). I get a reduction in NO3 and PO4, but nothing drastic. I don’t dose anything else but Alk and Cal. Doesn’t seem to cause any issues but the tank and sand appear cleaner. I do turn off the skimmer for 8 hours or so. It goes a bit nuts when turned back on, even with a water change.

So, in other words, it works for you.

 

[Dan_P]

I myself have noticed that after the first dose (half dose) of WA, it has plummeted my phosphate to zero within hours (Which I currently dose daily to maintain a reading of 0.03-0.1) and ever so slightly reduced nitrates, but not completely (which matches WA product marketing and @taricha 's findings) which is great.

WasteAway probably has nothing to do with the PO4 depletion.

This viewpoint is based on experiments (Inspired by @taricha ) I ran in which WA, added at the recommended dose to a sample of aquarium water, showed no signs of growth after 5 days.