Bacteria in a bottle, Myth or Fact

[Dr. Reef]

Update
Day 10

Side study: After cycle dosed 1ppm

1. Fritz Turbostart 900 (cold) show 0-0.25 ppm
2. Fritz Turbostart 900 (Room temp) 0-0.25 ppm


Main Study:

1. Bio spira refused after cycle shows 0 ppm
2. Microbelift shows 1 ppm
3 ATM colony shows 0-0.25 ppm
4. Fluval cycle shows 1 ppm
5. Control shows 0.75-1 ppm

From left to right
Fritz cold, Fritz rm temp, bio spira, microbelift, ATM colony and Fluval cycle

 

[Dr. Reef]

@brandon429
I am about to reset this study. well in 2 more days or so. I dropped 10 pellets of fish food in all 6 tanks. If ammonia starts to drop in the one's that are still 1ppm this will prove that they carry heterotorph bacteria.
Once this study is reset, i know that Fritz cold bottle will drop ammonia to 0 in 3-4 days. Soon as it hits 0 i will do a 100% water change and dose 1 ppm again and start monitoring reduction in ammonia and how long it takes.

 

[Dr. Reef]

Day 12:

So i think i have a smoking gun. My theory seems to be working.
I placed 15 pellets of fish food in Microbelift and Fluval tanks yesterday. Both tanks have been holding steady at 1 ppm ammonia with multiple bacteria dosing as per instructions on the bottles.
After putting 15 pellets of fish food in yesterday, if tank had no bacteria to process the added food, ammonia should go higher as the food breaks down or other scenario would be hetertorphic bacteria will take up carbon from the decomposing food and also utilize ammonia to basically break it down for their food. Well here are the results.
After 1ppm ammonia for 10-11 days, adding few pellets of food which added carbon to the tank tell me these bacteria are heterotorph in nature as ammonia is 0.25-0.5 range today.
I cant call this conclusive but i am 90% sure. I will have to perform this test with 6 tanks to be 100% positive.
1. MicrobeLift
2 Microbelift + food
3. Fluval cycle
4. Fluval cycle + food
5. Prodibio
6 Prodibio + food
This will give us a better understanding of what type of bacteria we have in these bottles.

Pic from today
Left to right Microbelift, Fluval

 

[cromag27]

Day 12:

So i think i have a smoking gun. My theory seems to be working.
I placed 15 pellets of fish food in Microbelift and Fluval tanks yesterday. Both tanks have been holding steady at 1 ppm ammonia with multiple bacteria dosing as per instructions on the bottles.
After putting 15 pellets of fish food in yesterday, if tank had no bacteria to process the added food, ammonia should go higher as the food breaks down or other scenario would be hetertorphic bacteria will take up carbon from the decomposing food and also utilize ammonia to basically break it down for their food. Well here are the results.
After 1ppm ammonia for 10-11 days, adding few pellets of food which added carbon to the tank tell me these bacteria are heterotorph in nature as ammonia is 0.25-0.5 range today.
I cant call this conclusive but i am 90% sure. I will have to perform this test with 6 tanks to be 100% positive.
1. MicrobeLift
2 Microbelift + food
3. Fluval cycle
4. Fluval cycle + food
5. Prodibio
6 Prodibio + food
This will give us a better understanding of what type of bacteria we have in these bottles.

Pic from today
Left to right Microbelift, Fluval

do any of these products instruct you to use a carbon source as part of the cycle process?

 

[Dr. Reef]

yes and no, I'll explain.
Do they instruct carbon source out right?, No.
where did i get this idea then? Yes in a way that these products ask us to dose and add fish immediately. With addition of fish, comes addition of food.
Even if you were to not feed the fish there will be waste which will add carbon to the tank. So this theory of Heterotorph vs nitrifying bacteria is getting more clearer to me.

 

[Dr. Reef]

Look what Microbe Lift says on their website about carbon:

MICROBE-LIFT/Nite-Out II comprises select microorganisms that are autotrophic – able to use carbon dioxide as the sole source of carbon – and are relatively slow growing, requiring specific conditions for optimum growth with typical cell divisions rates from 8 to 16 hours. Their performance and rate growth is impacted by the environmental parameters required for nitrification.

From BioconLab:

True nitrifying bacteria are strictly aerobic autotrophs. They can only use nitrogen from inorganic sources such as ammonia and nitrite. Nitrosomonas (ammonia-oxidizers) and Nitrobacter (nitrite-oxidizers) are the most common.

 

[Dr. Reef]

going to bleach/breakdown tanks for round 2 of the study. Stay tuned.

 

[AngryOwl]

yes and no, I'll explain.
Do they instruct carbon source out right?, No.
where did i get this idea then? Yes in a way that these products ask us to dose and add fish immediately. With addition of fish, comes addition of food.
Even if you were to not feed the fish there will be waste which will add carbon to the tank. So this theory of Heterotorph vs nitrifying bacteria is getting more clearer to me.

Just started a cycle myself.. looking to boost it with carbon dosing. Can you tell me how to proceed? How much vinegar or vodka would I dose? Should I add a shot glass of Titos? Maybe a chaser as well?

 

[Victoria M]

Dr. Reef, a while back we discussed briefly how would Fritz hold up in copper. I just wanted to share that I switched from Stability to Fritz #9 for my copper QT and my seachem badges have not onced moved towards the green zone. I am very close to two weeks now. just wanted to share this info.

 

[Dr. Reef]

@AngryOwl
With carbon dosing you will need to start slow. Maybe 2.5-5ml per day for weeks then increase.
Personally I never cared for carbon dosing as it left a sludge everywhere in the system which overtime clogged my pipes/tubes and in rare case slowes the impeller of pumps.
Maybe someone else that has more experience with carbon dosing can chime in.

 

[Dr. Reef]

Dr. Reef, a while back we discussed briefly how would Fritz hold up in copper. I just wanted to share that I switched from Stability to Fritz #9 for my copper QT and my seachem badges have not onced moved towards the green zone. I am very close to two weeks now. just wanted to share this info.

Great info. I have been struggling to come up with some way to test it and so far I have not been able to come up with a conclusive process to test this. Maybe others can help me find a way.

 

[User]

Great info. I have been struggling to come up with some way to test it and so far I have not been able to come up with a conclusive process to test this. Maybe others can help me find a way.

My QTs are cycled with biospira and I’ve never seen a blip on my badge at any point using copper power at 1.75ppm for 14 days, or after during GC and observation fit 2 more weeks

10g tanks with maybe 3-3.5” of fish (eg 2x 1.5” clowns) at a time

That was some run on sentence. My apologies

 

[lakai]

I estimate 12 days average across strains. CroMag says four.

I agree with CroMag that it takes 4days IF there is sand. The tank can fully handle up to 4ppm ammonia and bring it down to zero or near zero on its own in 8-12 hours. I've been able to replicate the same results across 3 smaller tanks monitoring with a seneye.

 

[brandon429]

Interesting it requires sand, as this is a surface area test as much as it is a bacteria vitality one. Glad we'll get to see results here

So you did two tests, one without sand. Everyone but me requires nitrite compliance to stamp a cycle complete, are you getting full nitrite compliance as well? (Both ammonia and nitrite comply in four days total=new substrates formerly dry/inactive, dosed with ammonia and bottle bac only for four days, then full water change, then redose 2 ppm add no bac, both params comply within 24 hours)

The reason the unnamed test is so fascinating is because it measures not just the vitality/activation level of the initial dose, but it measures how fast the extra surface area is actually colonized over four days. If there is literally no difference in metabolic activity every 24 hours between the two systems that'll be fantastic.

The side test here shows if the initial dose recommended is active enough/enough cells per dose to adhere to all surfaces without requiring time to multiply

If removing sand causes the backup, then it's a surface area issue failing the test and this is rare as sand surface area isn't required in typical reefing (in the sand rinse thread we remove whole beds at once, without ramp down, using this rule) Live rock and or siporax should be enough.

 

[MnFish1]

which would smash the whole “suspension cycle” claim.

Im not quite sure what the 'suspension cycle claim' is. @brandon429 It seems logical to me that when you add bacteria to a system that until it attaches to a surface it will still remove ammonia. If you add bacteria and take out all the water 10 minutes later - its not likely that the tank will cycle. If you do it an hour later it probably still won't work. to do this test properly - you'd need to do lets say 5 tanks where you put bacteria into all of Them on day 0 - and do a water change lets say on day 1, 2, 3, 4, and 5 or (1,3,5,7,9). My understanding is that (at least some) Bacteria rapidly can attach with no 'biofilm' and rapidly detach.

In any case - It seems like its an academic question - because whether the bacteria is 'working' while attached or 'detached' - its still working - And again Im not asking this to critique - I just want to understand the point.

 

[MnFish1]

To clarify I'm highly interested in how long it takes a brand new set of substrates+ tank water to be able to demonstrably reduce ammonia due to bottle bac additions, endure a full water change, and still reduce same ammonia without re dosing bottle bac

It's testing actual deposition times for bac, and more active strains should adhere faster as my guess.

We can dose active bottle bac to a cup of sw, input tiny amnt of ammonia, and watch that system reduce ammonia because of what's dosed in suspension only.

But it takes a real amount of surface area, and adhesion by added bac, to then endure a full water change and be able to repeat the process without dosing new bac

I really did try and condense it MN is that clearer

We can make good use in cycling threads the knowledge of finding out how fast surfaces area coated and activated by bottle bac, and this water change test is the best way to know. I estimate 12 days average across strains. CroMag says four.

There is a period just after bottle bac dosing where params are changed, but not due to active substrates. Then the system changes into real substrate cycling given time (?)
That little spread is new data awaiting for the hobby and useful as well if it doesn't wreck Dr Reefs original goals.

Yes - its clearer - Im just not sure that there is any such thing as 'substrate cycling' and 'suspension cycling'. Its well known that nitrifiers attach and detach and become free-swimming in nature and in our tanks. They do not just 'plop down' and never move. So - Im just not sure that this will tell anyone anything - except perhaps what 'percent' of bacteria are 'adhered' vs 'in suspension'.

 

[MnFish1]

So you did two tests, one without sand. Everyone but me requires nitrite compliance to stamp a cycle complete, are you getting full nitrite compliance as well? (Both ammonia and nitrite comply in four days total=new substrates formerly dry/inactive, dosed with ammonia and bottle bac only for four days, then full water change, then redose 2 ppm add no bac, both params comply within 24 hours)

The reason the unnamed test is so fascinating is because it measures not just the vitality/activation level of the initial dose, but it measures how fast the extra surface area is actually colonized over four days. If there is literally no difference in metabolic activity every 24 hours between the two systems that'll be fantastic.

The side test here shows if the initial dose recommended is active enough/enough cells per dose to adhere to all surfaces without requiring time to multiply

Sorry if I missed the answer - I haven't been getting notifications on my computer for any threads for a couple days

1. What is 'nitrite compliance?'
2. What evidence do you have that bacteria that are in suspension aren't just as actively metabolizing as those attached to substrate?
3. What evidence do you have that bacteria can't multiply when in suspension?

 

[hart24601]

This is really interesting stuff, really appreciate you doing these tests. You have done a great job. I am also curious about bacterial adhesion time which is similar to what I think Brandon is asking. Now clearly this isn’t going to be terribly important for someone setting up a tank as they are not likely doing large W/C but just something more personally interesting between products and how long the microorganisms take to attach to substrate.

 

[brandon429]

Good summary MN

I'm lucky to have him testing any aspect, if it's only fritz, the most likely to adhere in four days due to performance here, that's well enough.

It would be neat to see at least one contrast, the lowest or slowest performer here.

If it can't do both nitrite and ammonia at day four, post water change then I get to name it. We'll call it cro mag cycling phase if suspension cycling sounds too bad. After that point to be termed, cycle can be deemed complete.

The hallmark sign of the completed cycle is complete independence of oxidation ability to water changes...we're simply testing how fast that can be produced.

 

[mikeyn]

Good summary MN

I'm lucky to have him testing any aspect, if it's only fritz, the most likely to adhere in four days due to performance here, that's well enough.

It would be neat to see at least one contrast, the lowest or slowest performer here.

If it can't do both nitrite and ammonia at day four, post water change then I get to name it. We'll call it cro mag cycling phase if suspension cycling sounds too bad. After that point to be termed, cycle can be deemed complete.

The hallmark sign of the completed cycle is complete independence of oxidation ability to water changes...we're simply testing how fast that can be produced.

There are factors that determine how quickly the nitrifiers settle in and begin colonizing the media/surfaces. Temperature is a big factor, so cooler systems (like in Aquaculture, Lobster Systems, cold water exhibits in public aquaria) take longer. Water quality, does the water contains the things the nitrifiers need (alk, phosphates, trace elements, etc.) and nothing inhibiting them (Copper, etc.). Other things like flow, dissolved oxygen, amount of suitable media, etc. No two systems are exactly the same.