Nitrogen cycle - where are them nitrites and nitrates?!

[Ross Petersen]

Hey everyone,

I'm perplexed. I've got a small 10-gallon quarantine tank that I've set-up with a hang-on-the-back filter with filter pad, 5 ceramic bio-media balls as a substrate for nitrifying bacteria, and a small wave pump.

I can't seem to get the nitrogen cycle going. It's been 15 days (still early). However, 0 detectable nitrite and nitrates. This is peculiar given the following:
-I added ammonium chloride (Dr. Tim's) as per the instructions to get to 2 ppm ammonia
-I then dosed Microbacter7 following bottled instructions.
-After 10 days, ammonia was still at 2 ppm and there was no detectable nitrite.

Thinking the Microbacter7 bacteria might be a bad batch/overheated, I have supplemented with Seachem Stability from days 10-15. I also added a few mysis shrimp on day 10 of dosing at the recommendation of a few reputable reefers to provide a carbon source for the nitrifying bacteria.

Day 16 now and 2 ppm ammonia with no nitrite or nitrate.

Any ideas? Is it possible both nitrifying bottles are inactive? I purchased Seachem locally and Microbacter7 on Amazon.

Thanks,
Ross

 

[Quietman]

You should have seen some nitrites even without the Microbacter7. I personally didn't see any decrease in cycling time using the Mb7 over letting the naturally occurring bacteria populate. I did use the Dr Tim ammonia and it acted just like any other ammonia source (dead shrimp decay). There should also be more than enough nutrients in your system for a cycle without carbon adds.

The only things I can think are: more time, (16 days should have been enough to see nitrites though), test kit (always), antibacterial agent contamination (soap/filter pad), low temps might be retarding the bacterial life cycle...that last is a stretch.

 

[Idoc]

I've heard of cycles stalling if over 5ppm ammonia. Does your ammonia test kit max out at 2ppm readings possibly and there was actually much more ammonia in the system?

Without the MB7, you should have had something by now... even without it there should have been some movement toward nitrites by now.

 

[Quietman]

I've heard of cycles stalling if over 5ppm ammonia. Does your ammonia test kit max out at 2ppm readings possibly and there was actually much more ammonia in the system?

Without the MB7, you should have had something by now... even without it there should have been some movement toward nitrites by now.

Right...I've heard of that before...Too much ammonia can kill anything but not sure exact levels. 5 ppm is what i think the Dr Tim's bottle said.

 

[Lasse]

It sounds rather odd - first step should have started a long time ago (NH3/NH4 -> NO2) - both with and/or without bottled bacteria as other have stated. What do you use to test with?

Sincerely Lasse

 

[brandon429]

agreed. this is not a calibrated zero test, and if it becomes one, I bet we can get some proof of bacterial action. here's how to get it to work:

only ammonia factors-

change out all your water and take an ammonia readings at known zero (all clean new water)
take a pic of that color, its calibrated zero.

use dr tims liquid ammonium chloride and add into the tank at 1/4th doses, ultra tiny amounts, and take an ammonia measure and stop adding when the slightest color change for positive ammonia occurs.

not 2 ppm

not even 1 ppm, the first increment of change, about half ppm is right.

wait 24 hours and test ammonia, its back to calibrated zero color is the bet.

*it is fully ok to toil over nitrate and nitrite, hoping your 3x testing has no confounds or procedural errors to get a trusted reading (how are we calibrating the other two for accurate zero?) but you can also forego them, only ammonia matters.

The other two are always working when ammonia oxidation occurs, our ability to measure all three simultaneously is whats in constant challenge.

if your tank cant pass calibrated ammonia test after this long, then that bottle of bac was indeed sterile.

 

[Ross Petersen]

You should have seen some nitrites even without the Microbacter7. I personally didn't see any decrease in cycling time using the Mb7 over letting the naturally occurring bacteria populate. I did use the Dr Tim ammonia and it acted just like any other ammonia source (dead shrimp decay). There should also be more than enough nutrients in your system for a cycle without carbon adds.

The only things I can think are: more time, (16 days should have been enough to see nitrites though), test kit (always), antibacterial agent contamination (soap/filter pad), low temps might be retarding the bacterial life cycle...that last is a stretch.

Thanks. Temp has ran at 78-80F. I’m using a brand new Red Sea test kit.

I’m a high school chemistry teacher and going to go in tonight and use our Vernier electronic ammonia sensor. I have a hunch it’s going to read high ammonia, TBD.

Really wondering about antibacterial contamination, or some trace heavy metal in there. The quarantine tank did come with a cheap HOB that seemed to leach some plastic (I did a 100% water change at the get go to mitigate this).

So perplexing.

 

[Ross Petersen]

I've heard of cycles stalling if over 5ppm ammonia. Does your ammonia test kit max out at 2ppm readings possibly and there was actually much more ammonia in the system?

Without the MB7, you should have had something by now... even without it there should have been some movement toward nitrites by now.

Precisely. So at day 10 I did a 50% water change just in case the ammonia was too high. Ammonia has since dropped to below 2ppm. Still no nitrite via Red Sea.

RODI water via spectra-pure and using the basic instant ocean salt.

 

[Ross Petersen]

It sounds rather odd - first step should have started a long time ago (NH3/NH4 -> NO2) - both with and/or without bottled bacteria as other have stated. What do you use to test with?

Sincerely Lasse

Red Sea kit but going to verify tonight with a brand new $600 laboratory ammonia detector.

Thanks for the insights

 

[Quietman]

Whoa! High School Chemistry teachers don't muck around! Let us know what you find. Curious problem.

 

[Ross Petersen]

agreed. this is not a calibrated zero test, and if it becomes one, I bet we can get some proof of bacterial action. here's how to get it to work:

only ammonia factors-

change out all your water and take an ammonia readings at known zero (all clean new water)
take a pic of that color, its calibrated zero.

use dr tims liquid ammonium chloride and add into the tank at 1/4th doses, ultra tiny amounts, and take an ammonia measure and stop adding when the slightest color change for positive ammonia occurs.

not 2 ppm

not even 1 ppm, the first increment of change, about half ppm is right.

wait 24 hours and test ammonia, its back to calibrated zero color is the bet.

*it is fully ok to toil over nitrate and nitrite, hoping your 3x testing has no confounds or procedural errors to get a trusted reading (how are we calibrating the other two for accurate zero?) but you can also forego them, only ammonia matters.

The other two are always working when ammonia oxidation occurs, our ability to measure all three simultaneously is whats in constant challenge.

if your tank cant pass calibrated ammonia test after this long, then that bottle of bac was indeed sterile.

Thanks for the insights. I’m going into the lab tonight to get some serious accurate chemistry tests going (I’m a chemistry teacher).

I’m going to do some controls (as you’ve noted) and experiment with the gradient of sensitivity of the Red Sea kit as well.

Right now my leading hypotheses are:
-Bad bacteria (both bottles)
-Trace contamination from the cheap Petsmart quarantine tank I bought new.

Kind thanks

 

[Idoc]

Right now my leading hypotheses are:
-Bad bacteria (both bottles)
-Trace contamination from the cheap Petsmart quarantine tank I bought new.

Well, I'm not sure it's bad bacteria. Even without this, the bacteria should have colonized somewhat already naturally.

I would start to suspect a bad nitrate/nitrite test kit... or testing technique possibly. Look up your test kit on YouTube to ensure testing is being done precisely. I use Salifert for nitrate testing. I use Red Sea for other things, and they need to be done exactly as directed or else the results are off. I watched some youtube videos and figured out i was doing one of the tests (Mg) incorrectly.

 

[Ross Petersen]

Well, I'm not sure it's bad bacteria. Even without this, the bacteria should have colonized somewhat already naturally.

I would start to suspect a bad nitrate/nitrite test kit... or testing technique possibly. Look up your test kit on YouTube to ensure testing is being done precisely. I use Salifert for nitrate testing. I use Red Sea for other things, and they need to be done exactly as directed or else the results are off. I watched some youtube videos and figured out i was doing one of the tests (Mg) incorrectly.

Just ran ammonia and nitrite tests on RODI water: both are reading 0.

Just re-ran both on the quarantine tank. Ammonia about 1.2-2.0 and nitrite and nitrate 0. Followed each step religiously (and double-checked youtube).

Dr. Tim's ammonium chloride advertises to 2 ppm. The advertised concentration could be off; I just read a forum on this - apparently they printed old labels on some new bottles. Purely anecdotal.

The tank water is mildly cloudly. Bacterial bloom perhaps.

Either A) my ammonia was too high (I'd guess at 3.2 ppm until my 50% water change on day 10) or B) the bacteria - both bottles - are sterile. Or C) is it possible storing bacteria in the fridge is a bad move? I'm highly doubtful of this.

That plastic rock in the tank is apparently reef-friendly but I'm tempted to remove it as a triple safety.

 

[Idoc]

I'm baffled... maybe clear out the tank, clean it well with bleach or vinegar and start over?

Maybe the bacterial bloom is depleting oxygen or something that is taking away what is needed by the MB7 nitrifying bacteria?

I use biospira in my quarantine tanks without any problems. But, i don't cycle them before use... just fill with saltwater, add biospira a day or 2 before fish arrive... then add some biospira each day or 2 for the first week after fish arrive in the tank. I use a seachem ammonia alert badge and monitor it that way. Never a problem and i sometimes keep fish 3 months in quarantine.

 

[Ross Petersen]

I'm baffled... maybe clear out the tank, clean it well with bleach or vinegar and start over?

Maybe the bacterial bloom is depleting oxygen or something that is taking away what is needed by the MB7 nitrifying bacteria?

I use biospira in my quarantine tanks without any problems. But, i don't cycle them before use... just fill with saltwater, add biospira a day or 2 before fish arrive... then add some biospira each day or 2 for the first week after fish arrive in the tank. I use a seachem ammonia alert badge and monitor it that way. Never a problem and i sometimes keep fish 3 months in quarantine.

Baffled as well.

Biospira has outstanding reviews so I might give that a try.

Lots of water mixing so oxygenation problems unlikely.

I’m hesitant to change the water and clean the tank (right now) so will give it a week to play it’s course.

Thanks for the feedback

 

[Hermie]

maybe there was some bleach in the tank?

 

[juanrmattos]

I had a similar scenario with my 5 Gallon, cycled using Dry Rock, live sand with Aquaforest Bio S, Bio F (Which is basically bacter and a medium to feed the bacteria) and Life Bio Fil which is a biological media already loaded with dormant bacteria of the same strand as the Bio S; zero ammonia until day 10, bloom at day 15 (After adding Helix coralline bacteria boost the cycle), Diatoms on the rock and glass at day 17 and dots of green on the rock replacing the orange diatoms at day 22... Still as of today zeroing on Nitrites, Nitrates or Phosphates... Green is already receding and coralline freckles are starting to show up... I'm still expecting to see some bubble or hair algae some day soon... Don't want to jinx it but it seems is not going to get any worse (The rock remains super clean and both the Diatoms and the algae dots are so minor that my three snails take care of it fairly easy... Reef already has two tiny fish and one coral and they are all doing great.

I did some research on the subject at the beginning due to the lack of ammonia, the explanation was basically that the ammonia, the nitrites and nitrates were concentrated in the sand and rock and because the quantity & quality of the bacteria they were able to keep up with them in the water column (Same as the algae which is an indication of nitrites turning into nitrates and then dying off to complete the cycle). Right or wrong, it made sense to me.

Best of luck with your reef, I hope you cycle successfully

 

[Lasse]

Its a mystery - I do not trust NH3/NH4 test - by I usually trust NO2 tests. NO3 tests in the start is normally not needed - if there is NO2 in the water - NO3 tests will give a false reading. With as much as over 2 ppm NH3/NH4 added - you should have NO2 readings sometimes during this time, regardless addition of bacteria or not.

The only explanations I can see is that something in your water kill your nitrification bacteria or your oxygen levels is too low. Both of this explanations comes very low on the probability scale for me - but of course it can be true once in a while. The oxygen track demands that your oxygen levels is below 3 ppm for the first step (NH3/NH4 -> NO2). The second step demand more than 5 ppm oxygen to work satisfactorily.

About the Nitrospira genus (probably the active genus in biospira) - for long time it has only been seen as a nitrite oxidizer (NOB) - therefore not active in the first step of ammonia oxidizers (NH3/NH4 -> NO2) - AOB. However - lately it have been discovered that the Nitrospira genus contain species in the comammox (COMplete AMMonium OXidation) group and therefore some species/clades can work the full cycle. Because I do not know what Biospira contain - i can´t say if it is active in the first step, however IME - the first step normally start by itself. Its important to know that ammonia oxidation not only happens with help of bacteria - even Archean organism is active in this - normally refereed as ammonia-oxidizing archaea (AOA). The article linked describe the environmental conditions for ammonia oxidation very well.

Back to your problem. Because it is a quarantine tank - I suppose you have a working tank. If you have a filter in your main tank - I would clean it and take the waste in a bottle together with saltwater. Shake well and dose some every day. Put the bottle in the refrigerator. If you do not have any filter - just wash up some sand and take the sludge instead. But I want a clean start - not contaminate from my old water you maybe say. But in this case - your new fish will sooner or later meet the microorganisms of your DT and to introduce a limit amount of them in your quarantine may help their immune system and if they can´t stand your DT´s fauna of microorganism - you can treat already in the QT.

The trick with soil - outlined in this article - can work too

Sincerely Lasse

 

[JCOLE]

Is the Red Sea kit for Ammonia? If so, what kit do you use for Nitrate?

 

[juanrmattos]

Is the Red Sea kit for Ammonia? If so, what kit do you use for Nitrate?

The maturity kit includes all components to cycle your reef (Bacter, Nitro Bacter, No3.PO4.X and KH).