Bacteria ratios, putting a end to it!

[MnFish1]

How many of those thousands do we typically see in marine aquariums?

Correct, Heterotrophic bacteria can divide in 20 minutes and would outcompete the other bacteria.

This is an unanswerable question. I mean google - the number of bacteria species in a pond - I dont know. No one knows - even @AquaBiomics. Because I believe he measures certain bacteria. i.e. you cant measure every possible bacteria

 

[MnFish1]

I see we’re it went wrong, I didn’t account for the o2 depleting and the hole system become anoxic.
we’re do I go from here? Repeat the experience? The first 10 hours were still anaerobic.

no2 is between 0.6 and 0.8 ppm
Ammonia is at zero ppm
Phosphates are still at zero even after a additional 0.9 ppm after the 18 hour mark, we’re the phosphates going?

IMHO - you are trying to herd cats. It has nothing (and everything) i.e. - no one knows) - whether oxygen ws the issue - or surface area or anything else not specifically related to your experiment.

IMHO - You need to know what bacteria - is supposed to be doing the 'workload' - we do not. You should add bacteria - IMHO - at your next experiment. You also do not know if/whether the whole system became anoxic - in fact it likely did not unless you had an entirely cloudy tank. (which is the definition of a bacterial bloom).

 

[MnFish1]

I say that NO3 measurement "rebound" after an initial drop is possible for the O2 limiting reasons that others mentioned earlier.

Because 1mL vodka = .32 g ethanol = .17g Carbon, so 1mL vodka in 10L = 17mg/L Carbon. If half (ballpark guess) of that Carbon were oxidized to CO2, that would take 44 mg/L O2. Saltwater will have ~7 mg/L O2 max. So it's enough Carbon to deplete all the O2 like 6 times over. I'm guessing that the small pump may not keep up with the oxygen consumption once the bacterial bloom gets rolling.
If that happens, and O2 is gone in some part of the water while organic C remains, then NO3 could be reduced to NO2, and if that happens, then the NO3 test (Nyos?) may be very sensitive to the NO2 produced and generate a false high NO3. It was made even more likely since Sixty bumped the vodka up to 5mL.

(I've confused myself doing this sort of thing in a bottle of tank water before.)




But that is exactly what I do say. Tank water bacteria under high Carbon and low O2 de-nitrified NO3->NO2 and gave a false NO3 "increase"

Pull out an NO2 test kit to see if nitrite was in fact generated. Test for ammonia too, just for kicks.

I'm sorry this could be babble. there are multiple variables that are not accounted for. Since you admit yourself that you confused yourself before with doing 'this sort of thing' in a bottle of tank water before - I'm now confused. There is no evidence that this 10L experiment has low O2 IMHO. The problem here - is the tests being used are not designed to show increases from 10 to 13 or 20 to 25. This is not @Sixty_reefers issue - it's an issue of the tests being used.

 

[sixty_reefer]

IMHO - you are trying to herd cats. It has nothing (and everything) i.e. - no one knows) - whether oxygen ws the issue - or surface area or anything else not specifically related to your experiment.

IMHO - You need to know what bacteria - is supposed to be doing the 'workload' - we do not. You should add bacteria - IMHO - at your next experiment. You also do not know if/whether the whole system became anoxic - in fact it likely did not unless you had an entirely cloudy tank. (which is the definition of a bacterial bloom).

I see what you mean, the initial thoughts were mainly to try and illustrate that phosphates can be depleted using bacteria, I will be limited on my local resources to try and isolate the bacteria responsible for it although I believe it will already be known to many in the field.
At 10 hours into the test the tank wasn’t anoxic yet only a small haze was observed.

at 18 hours it’s clearly anoxic or in the region I may need to purchase a o2 test also

the bacteria responsible for this last bloom is unknown although is still consuming phosphates like crazy, I’ve added since the 18 hours mark 2.1ppm of phosphates and it fully depleted almost instantly. I would suspect that nitrifying bacteria couldn’t use this much phosphates

 

[MnFish1]

I see what you mean, the initial thoughts were mainly to try and illustrate that phosphates can be depleted using bacteria, I will be limited on my local resources to try and isolate the bacteria responsible for it although I believe it will already be known to many in the field.
At 10 hours into the test the tank wasn’t anoxic yet only a small haze was observed.

at 18 hours it’s clearly anoxic or in the region I may need to purchase a o2 test also

the bacteria responsible for this last bloom is unknown although is still consuming phosphates like crazy, I’ve added since the 18 hours mark 2.1ppm of phosphates and it fully depleted almost instantly. I would suspect that nitrifying bacteria couldn’t use this much phosphates

THIS is excellent - This is a bloom. . that was what I was getting at. If you had an oxygen source - you would have delayed this by xx hours - but - it would have inevitablely been the case. This the the bacterial stages I was talking about earlier - hopefully this will help

Bacteria - Reproduction, Nutrition, Environment | Britannica

Bacteria - Reproduction, Nutrition, Environment: Growth of bacterial cultures is defined as an increase in the number of bacteria in a population rather than in the size of individual cells. The growth of a bacterial population occurs in a geometric or exponential manner: with each division...

www.britannica.com

 

[sixty_reefer]

THIS is excellent - This is a bloom. . that was what I was getting at. If you had an oxygen source - you would have delayed this by xx hours - but - it would have inevitablely been the case. This the the bacterial stages I was talking about earlier - hopefully this will help

Bacteria - Reproduction, Nutrition, Environment | Britannica

Bacteria - Reproduction, Nutrition, Environment: Growth of bacterial cultures is defined as an increase in the number of bacteria in a population rather than in the size of individual cells. The growth of a bacterial population occurs in a geometric or exponential manner: with each division...

www.britannica.com

Thanks for the link, this is what brings more confusion to the table, nitrifying bacteria takes 16 hours to multiply I can’t see a possibility we’re I could provoke a nitrifying autotrophic bacteria bloom in 18 hours.

 

[MnFish1]

Thanks for the link, this is what brings more confusion to the table, nitrifying bacteria takes 16 hours to multiply I can’t see a possibility we’re I could provoke a nitrifying autotrophic bacteria bloom in 18 hours.

I do not think you could!!!!

 

[taricha]

I see we’re it went wrong, I didn’t account for the o2 depleting and the hole system become anoxic.
we’re do I go from here? Repeat the experience? The first 10 hours were still anaerobic.

no2 is between 0.6 and 0.8 ppm
Ammonia is at zero ppm
Phosphates are still at zero even after a additional 0.9 ppm after the 18 hour mark, we’re the phosphates going?

Yeah, simply for the sake of sanity of trying to measure NO3, you'll need to keep it well aerated. It's going to be hard to do that if you have a really high amount of Carbon.

Next time, tune back the vodka or crank up the aeration or both. Should keep the NO2 zero which will make NO3 tracking possible.

The PO4 is of course gone into biomass of that big cloudy bloom.

Here's what happened one time when I did some lowering of high NO3 and PO4 with vodka.

I started with a gallon of tank water, added Fritz F/2 media at half strength and half a salifert scoop of crushed fish flake then bubbled the result for a day or two to get the water nutrients semi-stable.

Starting nutrient level: 51ppm NO3, 2.61ppm PO4
I then split it into 3 bottles that I bubbled continuously in the dark. Control got nothing, 2nd got double recommended doses of Waste Away, and the 3rd bottle got vodka that was 1/6th of the WA (approximately carbon equivalent based on earlier O2 consumption measurements)

So here's what it looks like

Stars indicate which days WA and Vodka were added.
The control became more Carbon Limited around day 5 and nutrient reduction slowed a lot.
The Waste Away and Vodka treatments behaved really similarly to each other. In Phosphate, after day 6, they stayed within about one test error of each other, though the WA was almost always the lower one.
In nitrate, the difference was a little more clear - WA was consistently lower and became (and stayed) zero/undetectable on days 11&12.

and here's the N/P atom ratios that were generated from these measurements.

So I split the mole ratios of N, P, and C reductions into before and after day 5.

	N/P drop			C/P drop		C/N drop
	Control	W.A.	Vodka	W.A.	Vodka	W.A.	Vodka
Day 0-5	25.0	26.4	24.9	52	53	2.0	2.1
Day 5-12	31.0	162	135	1870	1430	11.5	10.6

Caveat on the C ratios, I don't know how much available C was in the samples to start with. I calculated based on the C that I added. So for days 0-5 there's way more C consumed within the sample than just what I added. But after day 5, probably the Carbon I added to the bottles was most of the carbon available.

I got an initial N/P ratio of about 25:1 for the decrease of NO3 and PO4 with nothing added, vodka added, (or Waste Away as carbon) for the first 5 days.

Days 5-12 the ratios of N/P consumption changed a lot.

 

[sixty_reefer]

Yeah, simply for the sake of sanity of trying to measure NO3, you'll need to keep it well aerated. It's going to be hard to do that if you have a really high amount of Carbon.

I’m going away for the weekend hence the overdosing, with more time just a drop of vodka would of done the job better although I tough if I was to create a bloom the main goal would of been shown as the bacteria multiply phosphates would of go down in a way that could be visualised with my current trst

Next time, tune back the vodka or crank up the aeration or both. Should keep the NO2 zero which will make NO3 tracking possible.

you right I’m my aquarium I just dose 2 ml per 100 litres to control phosphates

The PO4 is of course gone into biomass of that big cloudy bloom.

Here's what happened one time when I did some lowering of high NO3 and PO4 with vodka.


and here's the N/P atom ratios that were generated from these measurements.


I got an initial N/P ratio of about 25:1 for the decrease of NO3 and PO4 with nothing added, vodka added, (or Waste Away as carbon) for the first 5 days.

Days 5-12 the ratios of N/P consumption changed a lot.

I agree the real ratio for bacteria would only be found on a continuous and prolonged test, the first few days would always show a greater increase although there is a catch with my test and yours that needs to be addressed. You only added phosphates and nitrates at the beginning of the experiment, to fully mimic a aquarium environment a daily dose of nitrogen and phosphorus would have to be added to mimic the daily nutrients import added by food and fish waste to our systems. Would you not agree that if you had a daily import of nutrients to a systems that you would have to increase the amount of waste away or vodka to create a balance in residual? Not necessarily taking them dow to zero as that’s not the goal in our hobby, the goal is to have nitrates and phosphates at a constant residual. A experiment carried out this way would show that the vodka (carbon) is affecting directly the amount of residual nitrates and phosphates. It would also show that you would need to keep a greater number of bacteria available in the tank to keep that balance that could be similar to day 1-5 in your test.
To complicate things even further if the phosphates and nitrogen sources were randomly chosen you could observe during the test I just referred a increase or decrease on the residual in one or another the only way to achieve balance in a test like this would of been by manipulating the amount of nitrogen or phosphorus added to the system.

eventually this is we’re I want to be with this sort of test as this is what I wrote, unfortunately I am having problems as some folks don’t understand that wend you manipulate Carbon you can reduce nitrates and phosphates. Because most only read into nitrifying bacteria they don’t understand that this is possible to do fairly easily. Wend I speak of fixing ratios in other threads I am referring at adjusting nutrient to create a balance.

 

[jrmailo]

While outside the scope of this experiment, I would be interested in seeing a DNA sequencing identification of the vodka-induced bacteria. This would generally require a pure culture for precise identification, and here I would assume that your bacterial bloom only contains one or a few (at most) of dominant bacteria strains that took advantage of the organic C availability.

It would also be interesting to see if the pre-dominant growth of the bacterial specie(s) change with different organic C dosing (vodka/vinegar/glucose/etc).

I think that the identification of these bacteria will give us a better understanding of the ratio of N and P reduced via organic C dosing by bridging our understanding with existing scientific literature.

 

[sixty_reefer]

While outside the scope of this experiment, I would be interested in seeing a DNA sequencing identification of the vodka-induced bacteria. This would generally require a pure culture for precise identification, and here I would assume that your bacterial bloom only contains one or a few (at most) of dominant bacteria strains that took advantage of the organic C availability.

that would be ideal, I need to research if there is facilities similar to aquabiomics in the United Kingdom to be able to identify at list the species involved in the bloom

It would also be interesting to see if the pre-dominant growth of the bacterial specie(s) change with different organic C dosing (vodka/vinegar/glucose/etc).

I would imagine that it would I have read that glucose and molasses would be more effective on denitrification.

I think that the identification of these bacteria will give us a better understanding of the ratio of N and P reduced via organic C dosing by bridging our understanding with existing scientific literature.

The identification of the species could be beneficial to the hobby, and we all should have them in our systems although I read that most of the pelagic bacteria can be removed via skimming, finding a balance here could be difficult

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

 

[taricha]

I agree the real ratio for bacteria would only be found on a continuous and prolonged test, the first few days would always show a greater increase although there is a catch with my test and yours that needs to be addressed. You only added phosphates and nitrates at the beginning of the experiment, to fully mimic a aquarium environment a daily dose of nitrogen and phosphorus would have to be added to mimic the daily nutrients import added by food and fish waste to our systems.

I agree that the bacterial removal of NO3 and PO4 from tank water (and the N/P ratios) will depend greatly on the conditions present. And experiments that mimic aquarium conditions more closely will generate more relevant data.

 

[sixty_reefer]

I agree that the bacterial removal of NO3 and PO4 from tank water (and the N/P ratios) will depend greatly on the conditions present. And experiments that mimic aquarium conditions more closely will generate more relevant data.

The Problem I see is how important would that data be?

for example in one experiment were 3ppm nitrogen and 0.001 ppm phosphorus daily import could generate a balance at 5ppm nitrates and 0.01 phosphates with a X amount of vodka

and in other experience I could use 6ppm nitrogen and 0.002 ppm phosphorus daily import and generate also a balance at 5ppm nitrates and 0.01 phosphates with a Y amount of vodka

Could I generate a useful ratio using this two anecdotal experiments? We’re the daily import is different and the same residual balance is achieved?

I would imagine that I would be using more bacteria and vodka in the experiment with more nitrates and phosphates added daily although I’m finding confusing how I could create a useful ratio under different conditions of import that will have similar residual as a balance.

Do I need to use my daily import of nutrients to generate a useful ratio or would I have to use the residual nutrients of 5ppm and 0.01 phosphates to create a useful ratio in this anecdotal results of the possible experiments?

@MnFish1 and @Dan_P id like your views on this question also please

 

[Reefahholic]

I see we’re it went wrong, I didn’t account for the o2 depleting and the hole system become anoxic.
we’re do I go from here? Repeat the experience? The first 10 hours were still anaerobic.

no2 is between 0.6 and 0.8 ppm
Ammonia is at zero ppm
Phosphates are still at zero even after a additional 0.9 ppm after the 18 hour mark, we’re the phosphates going?

Put an air-stone in there. Prolly won’t be enough if the dosing is too aggressive. Why not run it out longer instead of trying to get instant results. Maybe add a skimmer too and let it overflow.

 

[taricha]

Sixty, fundamentally you want to know how much N/P (as no3 and po4) that carbon dosing removes from the water.
You can't anticipate every system, so if I were you, I'd run a test or two at reef plausible parameters, and if I'm lucky, maybe vodka dosing in other systems will consume nutrients similarly - to within a factor of two. That's probably as useful as the data can get.

 

[sixty_reefer]

Put an air-stone in there. Prolly won’t be enough if the dosing is too aggressive. Why not run it out longer instead of trying to get instant results. Maybe add a skimmer too and let it overflow.

I only had one day available to try and do some preliminary findings before I shootout for the weekend, will most likely pick it up again from Monday, restart the test with a more balanced doses for the system size.

 

[Reefahholic]

I got to a point that in tank experience they do drop just after the phosphates bound to the substrate and rock are depleted, I think this is something I need to know whatever way it goes.

What did you find that depleted the PO4 bound to the calcium carbonate surfaces and how long did that take? I have a very interesting story to share on that particular subject that will leave you scratching your head or be a huge piece to the puzzle.

 

[sixty_reefer]

What did you find that depleted the PO4 bound to the calcium carbonate surfaces and how long did that take? I have a very interesting story to share on that particular subject that will leave you scratching your head or be a huge piece to the puzzle.

The phosphates in my system were bound to a different surface, I used aquatic compost to start my filter and there was phosphates bound to it, it took me 2.6 lbs of captive phos to remove 0.3 ppm of bound phosphates in a 26 gallon with no liverock over a 2 week period. I am curious about your experience.

 

[sixty_reefer]

Sixty, fundamentally you want to know how much N/P (as no3 and po4) that carbon dosing removes from the water.
You can't anticipate every system, so if I were you, I'd run a test or two at reef plausible parameters, and if I'm lucky, maybe vodka dosing in other systems will consume nutrients similarly - to within a factor of two. That's probably as useful as the data can get.

Thank you, where I’m getting is that the practicality of findings that ratio wouldn’t be very useful, the only useful thing I can see here is the fact that some bacteria in our tank will remove phosphates by assimilation from our tanks. The research you shared before also illustrate this.

 

[Reefahholic]

The phosphates in my system were bound to a different surface, I used aquatic compost to start my filter and there was phosphates bound to it, it took me 2.6 lbs of captive phos to remove 0.3 ppm of bound phosphates in a 26 gallon with no liverock over a 2 week period. I am curious about your experience.

Long story short (2 story’s)

First let me say I’ve typically had PO4 leech for up to 6 months, but never once have I personally had a system bind it.


This system has been binding my PO4 for 5+ months now. I’m currently dosing 0.14 ppm PO4 daily. Yes…that “isn’t” a typo! .14 ppm daily. It’s pretty crazy! The rock is sucking it up like Kool-Aid.


Here’s my only logic as to why I think this is happening:

The rock (Marco dry rock) was cured (in a Brute can) for just over 1 year. The first 6 months I treated it like a reef tank. Fed it, dosed it, kept the N&P around 5 and .05 ppm. I added multiple species of bacteria, live sand, pods, coralline algae, etc.

Here’s where I think it went wrong;

The second 6 months I got hit hard in the hospital with Covid. I’m a frontline healthcare worker. So obviously I was coming home beat down day in and day out. I let it go to poop. Quit testing, didn’t dose it, no more N&P, no more nutrition,
Nothing.

I think the second six months the Bacteria completely depleted the N&P and maybe even ate into the rock surface a bit tying to get food. Not sure what happened, but it had to be something similar, because the rock is taking so much PO4 it’s insane. No matter what I dose it’s sucking it down like it wants to forever drink. I’m sure it will hit a saturation point soon, but wow…I thought I have seen it all until now. I thought this tank would bypass every other system because I cured the rock for so long, and it probably would have had I not added that bacteria.

It’s either that, or I got some completely F ***ed up rock.

What do you guys suspect is going on here.?