Fritzyme 9 modified cycle

[Garf]

Thought I’d go “live bacteria” route for cycling a QT as the wife is itching to get a few fish in her tank. The most available bacteria by me was Fritzyme 9, so that’s what I went with.

Tank, ceramic rock, heater, Circulation pump were all bleached for 24 hrs then allowed to dry for 24 hrs prior to start up.

1) 40 liters of RODI / saltwater (Tropic Marin Classic) was added to the tank at 30ppt.
2) Tank heated to 26C
3) Added 50mls white vinegar (tank pH at 7.4 to 7.6ish), a bit vague but didn’t fancy messing with pH probes.
4) Added TriSodium Phosphate to 0.5ppm tested Hanna/API combo
5) Added Fritzyme 9 (8 ounce) as suggested by fritz for new startups.
6) Added 2ml Dr Tim’s ammonia.

The TAN according to API was good enough on 2ppm (photo is next to a freshwater card so you can ignore the color chart)

Tank tested this morning, a little over 18 hrs after bacterial addition has showed a 50% reduction in NH3/NH4, an increase in Nitrite and pH. In particular pH is now above 7.8
The Seachem ammonia alert has only now started to indicate a slight tinge, presumably due to the pH increasing, but still safe.

Tank is cloudy.

Fuzzy pics;

 

[Garf]

22 hrs;

 

[MnFish1]

Nice - it will be interesting to see what happens today. BTW - why the vinegar? Thats one thing thats not particularly 'standard' AFAIK. Additionally, why the phosphate?

 

[Garf]

why the vinegar?

Figured these are heterotrophic and required a carbon source, I think this is evidenced by the bloom.

why the phosphate

Also figured heterotrophs require phosphate and didn’t want to deprive them of such. As it happens though, I can’t detect any phosphate consumption.

26 hrs;

 

[Garf]

Took the opportunity to test my reef tank at the same time, it’s slightly different to the results I’m getting on the QT;

I’m making the assumption that the bacterial bloom in the QT sample is messing with the test somehow.

 

[flashsmith]

I used fritz 9 for several tanks over the last 3 years or so. Usually add livestock within 24-36 hours. Never an issue. I trust it to the point that I don't even test anymore. Instacycle works for me.

 

[Garf]

Been reading up on what actually causes bacterial settlement and although I don’t claim to fully understand all of it, it appears mainly driven by stressors, such as running out of nutrients, extreme pH, UV light etc;

Beyond Risk: Bacterial Biofilms and Their Regulating Approaches

Bacterial biofilms are complex surface attached communities of bacteria held together by self-produced polymer matrixs mainly composed of polysaccharides, secreted proteins, and extracellular DNAs. Bacterial biofilm formation is a complex process and can be described in five main phases: (i)...

www.frontiersin.org

 

[MnFish1]

Figured these are heterotrophic and required a carbon source, I think this is evidenced by the bloom.

Also figured heterotrophs require phosphate and didn’t want to deprive them of such. As it happens though, I can’t detect any phosphate consumption.

26 hrs;

Actually, you I do not believe are correct. Fritz 9 - is designed to be used without carbon - as the experiments from Dr. Reef showed. Thus you're right - you probably induced a bloom because of the carbon - but there is no reason to think that bloom was the bacteria in Fritz 9 (obligate nitrifiers do not have that quick of a doubling time). As to the phosphate consumption - my guess is that the tests are not sensitive enough to see it.

 

[MnFish1]

Took the opportunity to test my reef tank at the same time, it’s slightly different to the results I’m getting on the QT;

I’m making the assumption that the bacterial bloom in the QT sample is messing with the test somehow.

This would be at most a 0.25 ammonia - and probably a 0 - depending on lighting. Which test result is this? (Your tank or the QT tank - it looks like the tank)? I would read this a 0 ammonia and 0 nitrite.
PS - when you're testing your tank for the experiment -it's important to test at the same time each day.

 

[Garf]

obligate nitrifiers do not have that quick of a doubling time

Indeed

it looks like the tank

i wrote TANK on the pic from the display so I didn’t get confused, so that’s the display tank.

 

[taricha]

Thank you, @Garf this is a nice, fast result that interests me.
Since you posted it in the research and experiments forum, I have an unnecessary request.
Could you now add the same dose of ammonia again, this time without the vinegar.
I am interested in whether your initial combination of ammonia, phosphate, and vinegar quickly scaled up the cycling population, and now they are primed to go, or if the work was being done by heterotrophs that lose their effectiveness once the carbon source - vinegar - is exhausted.

 

[Garf]

Thank you, @Garf this is a nice, fast result that interests me.
Since you posted it in the research and experiments forum, I have an unnecessary request.
Could you now add the same dose of ammonia again, this time without the vinegar.
I am interested in whether your initial combination of ammonia, phosphate, and vinegar quickly scaled up the cycling population, and now they are primed to go, or if the work was being done by heterotrophs that lose their effectiveness once the carbon source - vinegar - is exhausted.

Done.
I’ve added another 2 ppm ammonia to the QT and will retest in about 12 hrs.

 

[Garf]

I’ve been googling furiously to try and explain the fast nitrite reduction with relation to heterotrophs, unsuccessfully, so far. Any guidance welcome
I hope @Lasse isn’t watching his windmills again.

 

[Garf]

Relevant or not?

“In addition to chemolithoautotrophic nitrite oxidation, some NOB can grow heterotrophically (Bock, 1976; Starkenburg et al., 2008), and nitrite oxidation by some Nitrobacter and Nitrospira species is accelerated when the medium is supplemented with simple organic compounds like formate (Bock, Koops, Moller, & Rudert, 1990; Daims, Nielsen, Nielsen, Schleifer, & Wagner, 2001; Watson et al., 1986).

Nitrite Oxidizing Bacterium - an overview | ScienceDirect Topics

www.sciencedirect.com

 

[brandon429]

I like how controlled this setup is against photosynthetics vs bacterial processing of test load ammonia. how controlled the surface area is compared to a reef tank where surface area is well overdone six times over... from these doses and resolve rates we're able to advise others on quarantine setups simply by them copying the general layout and contact areas in the flowpath here, even if they don't have as good of control over api. a bare bones new setup like this is transferable to other people's copied setups= handy.

 

[taricha]

With that high C/N ratio, there's a bunch of possibilities.
"Simultaneous heterotrophic nitrification/denitrification" is a thing that comes up in research papers.
Also, you grew a bunch of biomass, all forms of N are fair game if that much C is around.

 

[Garf]

I like how controlled this setup is against photosynthetics vs bacterial processing of test load ammonia. how controlled the surface area is compared to a reef tank where surface area is well overdone six times over... from these doses and resolve rates we're able to advise others on quarantine setups simply by them copying the general layout and contact areas in the flowpath here, even if they don't have as good of control over api. a bare bones new setup like this is transferable to other people's copied setups= handy.

I can confirm the tank only receives a tiny amount of ambient light, photo follows of ammonia alert 3 hrs after adding 2 ppm ammonia, (taken with the aid of a torch ) . Hasn’t budged.

you grew a bunch of biomass,

i think everyone should experience a bacterial bloom, lol

 

[MnFish1]

Thank you, @Garf this is a nice, fast result that interests me.
Since you posted it in the research and experiments forum, I have an unnecessary request.
Could you now add the same dose of ammonia again, this time without the vinegar.
I am interested in whether your initial combination of ammonia, phosphate, and vinegar quickly scaled up the cycling population, and now they are primed to go, or if the work was being done by heterotrophs that lose their effectiveness once the carbon source - vinegar - is exhausted.

heterotrophs do not just die when carbon is gone. they - depending on the environment can be Dormant. Pseudomonas, for example can live in bottles of betadine (the iodine based antiseptic). You cannot tell much from this experiment as to the population of bacteria.

 

[MnFish1]

I can confirm the tank only receives a tiny amount of ambient light, photo follows of ammonia alert 3 hrs after adding 2 ppm ammonia, (taken with the aid of a torch ) . Hasn’t budged.

i think everyone should experience a bacterial bloom, lol

I think you are ascribing things to things that do not relate - and IMHO - you have too many variables to tell anything except - that your tank can process ammonia quickly (which is good)- IMHO - nothing can be judged as to what bacteria, etc are doing the job. One issue with experimental design - promoted in this forum - is giving your thesis - and performing the experiment - as compared to adding multiple more variables after the fact.

 

[Lasse]

I´m awaiting your next test result. If you read my 15 steps - you will see that I suggest a PO4 dose too. (not vinegar though) Even the autotroph nitrificators need PO4 !

Generally adding of DOC will favour heterotrophs and disfavour the autotrophs. However the second stage bacteria - when they kick in - will convert produced NO2 quickly into NO3. I have seen 1- 2 mg/L NO3 been converted into NO3 in less than 10 hours when it kicks in

Sincerely Lasse