Bacteria in a bottle, Myth or Fact

[Dr. Reef]

Ah I see Dr. Reef came up with a pretty similar idea. Great minds...

This will be interesting to see!

Lol yes great minds think alike...
Now that i have pretty much come to an end of main studies testing 8 bacteria manufacturers, I have a pretty good idea which bacteria is true nitrifying bacteria, which one is heterotorph. Its time to play with other ideas soon.

 

[RamsReef]

What if it was bacteria that survived on solvent fumes?

 

[lakai]

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.

bacteria is already attatched to some type of substrate which is the reason I believe for the cloudiness. Allowing it to flow through your system and settle is important. The ammonia oxidizing bacteria in the bottle is starved and will immediately start working once they have a source of food. These bacteria are colonies and adhesion is vital to long term survivability. Again bottled bac is already adhered to substrate but needs to settle and to be able to grow onto new substrate.

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

We all know by now which one will perform the quickest so test them against each other doesn't make sense.
I think the only test that matters is how long it takes for each bottled product to be able to sustain livestock using the same dosage of each of the products. This test can even be done in a test tube or bucket of saltwater it doesn't matter as its can scale. The most fair way is all tanks setup with the same amount of substrate, flow, temperature.

1. Dose each tank to 2ppm using pure ammonium chloride, verify in real-time using seneye. ( Make sure it reads 0 nh3 before dosing)
2. Shake each bottle for 30 seconds and dose exactly 10mL to tank. Do not dos per mfg recommendation.
   
3. Monitor ammonia until it goes down to 1ppm ( This way we know it is active and working. Not waiting for it to read zero is so that the bacteria is allowed to keep feeding so that it can keep growing. We can expect certain bottles to take longer before.)
4. Once NH3 reads 1ppm, Dose pure ammonia chloride to 4ppm and add another 10mL of bacteria.
5. Wait until Nh3 level reads .5 ppm (This tells us the ammonia oxidizing bacteria has multiplied enough to be able to convert more ammonia than any livestock could create anytime. Should take anywhere between 12-24 hours.)
6. Dose ammonia chloride to 4ppm again. (This time we see how long it takes to go from 4ppm to zero.)
7. Once nh3 reads zero. Test for nitrite. (By this time we should see the tank can handle 4ppm ammonia and take it down to zero in a reasonable time 3-5 hours)
8. Test until nitrite reads zero or near zero.

Thats it. Once NH3 and Nitrite both read zero you could do a water change then dose another 4ppm ammonium and see how long until nitrite reads zero and confirm how long it takes from ammonia > Nitrate but it doesn't really matter what the nitrate reading is at this point because we know the tank is ready for fish now.

 

[MnFish1]

In my studies that I have already conducted, Fritz Turbostart 900 has always cycled my 5 gal tank in 3 days, which tells me that bacteria was probably out of water column within first 24 hrs as it would take 15-18 hrs for initial bacteria dose to double its numbers and start oxidizing till 48 hrs later it can completely consume 1 ppm or 2 ppm and in fact even 8 ppm within 3 days to 0.

I have a few ways to do this like @brandon429 wants.
These 3 will be 3 separate studies.

1. Day 1 dose ammonia and bacteria , cycle 3 days later, do 100% water change and redose ammonia.

2. Day 1 dose ammonia and bacteria, day 2 do 100% water change and redose ammonia.

3. Day 1 dose ammonia and bacteria, 24 hrs do 100% water change and redose ammonia.

From doing 100% water change on day 1, day 2 and day 3 on 3 separate studies can narrow it down to when the bacteria fell off the water column.

disagree - there is no way to tell if the bacteria have adhered to something - before they start metabolizing things...

 

[MnFish1]

bacteria is already attatched to some type of substrate which is the reason I believe for the cloudiness. Allowing it to flow through your system and settle is important. The ammonia oxidizing bacteria in the bottle is starved and will immediately start working once they have a source of food. These bacteria are colonies and adhesion is vital to long term survivability. Again bottled bac is already adhered to substrate but needs to settle and to be able to grow onto new substrate.



We all know by now which one will perform the quickest so test them against each other doesn't make sense.
I think the only test that matters is how long it takes for each bottled product to be able to sustain livestock using the same dosage of each of the products. This test can even be done in a test tube or bucket of saltwater it doesn't matter as its can scale. The most fair way is all tanks setup with the same amount of substrate, flow, temperature.

1. Dose each tank to 2ppm using pure ammonium chloride, verify in real-time using seneye. ( Make sure it reads 0 nh3 before dosing)
2. Shake each bottle for 30 seconds and dose exactly 10mL to tank. Do not dos per mfg recommendation.
   
3. Monitor ammonia until it goes down to 1ppm ( This way we know it is active and working. Not waiting for it to read zero is so that the bacteria is allowed to keep feeding so that it can keep growing. We can expect certain bottles to take longer before.)
4. Once NH3 reads 1ppm, Dose pure ammonia chloride to 4ppm and add another 10mL of bacteria.
5. Wait until Nh3 level reads .5 ppm (This tells us the ammonia oxidizing bacteria has multiplied enough to be able to convert more ammonia than any livestock could create anytime. Should take anywhere between 12-24 hours.)
6. Dose ammonia chloride to 4ppm again. (This time we see how long it takes to go from 4ppm to zero.)
7. Once nh3 reads zero. Test for nitrite. (By this time we should see the tank can handle 4ppm ammonia and take it down to zero in a reasonable time 3-5 hours)
8. Test until nitrite reads zero or near zero.

Thats it. Once NH3 and Nitrite both read zero you could do a water change then dose another 4ppm ammonium and see how long until nitrite reads zero and confirm how long it takes from ammonia > Nitrate but it doesn't really matter what the nitrate reading is at this point because we know the tank is ready for fish now.

Why 2 ppm? if you added a tang for example - to a 100 gallon aquarium - nothing else - how long would it take to get to 2 ppm? If you added 2 clownfish to a 100 gallon tank - how long would it take to get to 2 ppm? What is the rationale for this number - except for dr. Tim saying - start at 2 ppn. Which is ridiculous *(unless he has data to suggest that level) - which isnt published. In the mean time - No one knows what they are doing with regards to cycling tanks IMHO

 

[Victoria M]

Why 2 ppm? if you added a tang for example - to a 100 gallon aquarium - nothing else - how long would it take to get to 2 ppm? If you added 2 clownfish to a 100 gallon tank - how long would it take to get to 2 ppm? What is the rationale for this number - except for dr. Tim saying - start at 2 ppn. Which is ridiculous *(unless he has data to suggest that level) - which isnt published. In the mean time - No one knows what they are doing with regards to cycling tanks IMHO

Um, except Dr. Reef. He now Knows what he is doing. As do we all at this point, IMO, anyway. I have seen enough evidence to switch to Fritz. Even the Fritz 9 has been a solid performer in my last QT, even with copper.

 

[Dr. Reef]

I dont want to call it till last of my tests are done but yes you are right @Victoria M
If someone was to cycle a tank in a hurry
Fritz Turbostart 900 is the best.
If you have time to cycle then Dr Tim or Bio Spria will work just fine but would need to be shaken really well and try to pull fluid from the bottom of the bottle.
These 3 products are the only ones that have worked so far in a clean new setup.
Rest 5 will work if there were fish or some carbon source.

 

[MnFish1]

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.

This does not mean that these products contain non-nitrifying bacteria (or am I misunderstanding your point). Nitrosomonas for example also can use CO2 as their sole carbon source - so it anything both of these products are suggesting that they contain a nitrosomonas type bacteria...

"Nitrosomonas europaea can obtain the carbon that it needs to grow by getting it from the atmosphere via "carbon fixation": converting carbon in a gaseous form into carbon bound up in organic molecules"

 

[MnFish1]

Um, except Dr. Reef. He now Knows what he is doing. As do we all at this point, IMO, anyway. I have seen enough evidence to switch to Fritz. Even the Fritz 9 has been a solid performer in my last QT, even with copper.

What (we all know) - is that using Fritz Turbo at a 1 ppm and 2ppm and 8 ppm ammonia levels is that ammonia drops within 1-2 days to 0. As @Lasse and @brandon429 have said. we don't know how this translates to a tank situation. All that has been 'proven' IMHO - is that Fritz Turbo can perform well in this test - and that some of the other products work 'a little' - but not that well. It is an extremely interesting experiment - and it certainly proves the question that started the thread: Bacteria in a bottle myth or fact.

When I was saying 'we dont really know what's going on in a cycle' - I meant that for example - I have used seachem stability many times - in clean sterile aquaria - and had no issues with fish loss/poor cycling, etc. I dont know why that it doesn't work here - but works in a natural aquarium - several others have posted similar results. Many people have stated that non-cooled bacteria in a bottle doesnt work as well as 'cooled bacteria' - yet Fritz 9 is non-cooled.

All I was trying to say is that the experiments bring up many more interesting questions yet to be answered - as @Dr. Reef has said. Thanks again to him for doing all the work.

 

[MnFish1]

I'm not sure the copper thing is that important - but - I agree with @cromag27 use the instructions with the products. ie.

1 tank - water/filter/salt/bacteria/ammonia
1 tank water/filter/salt/bacteria/(copper dosed to daily dose levels with copper test kit)/ammonia
1 tank water/filter/salt no bacteria/ammonia
1 tank water/filter/salt/no bacteria/no ammonia

Sorry that contained a mistake. Should have been

Tank 1. 1 tank - water/filter/salt/bacteria/ammonia
Tank 2 1 tank water/filter/salt/bacteria/(copper dosed to daily dose levels with copper test kit)/ammonia
Tank 3. 1 tank water/filter/salt no bacteria/ammonia
Tank 4 1 tank water/filter/salt/no bacteria/no ammonia/ but add copper to daily dose levels with proper test kit.

Tank 1 is the control for tank 2
Tank 2 is the 'test tank'
Tank 3 is the standard ammonia control.
Tank 4 is to be certain that copper alone doesnt affect ammonia test kits (unlikely)

 

[Lasse]

Nitrosomonas europaea can obtain the carbon that it needs to grow by getting it from the atmosphere via "carbon fixation": converting carbon in a gaseous form into carbon bound up in organic molecules"

Where have you got this information from - I ask because I´m not sure it is right if you see it like the way plants take up inorganic carbon. IMO - the inorganic carbon that nitrifiers use comes from HCO3 or CO3. It means an indirect uptake of CO2.

Sincerely Lasse

 

[MnFish1]

Where have you got this information from - I ask because I´m not sure it is right if you see it like the way plants take up inorganic carbon. IMO - the inorganic carbon that nitrifiers use comes from HCO3 or CO3. It means an indirect uptake of CO2.

Sincerely Lasse

https://genome.jgi.doe.gov/portal/niteu/niteu.home.html

IN the last paragraph

 

[Victoria M]

What (we all know) - is that using Fritz Turbo at a 1 ppm and 2ppm and 8 ppm ammonia levels is that ammonia drops within 1-2 days to 0. As @Lasse and @brandon429 have said. we don't know how this translates to a tank situation. All that has been 'proven' IMHO - is that Fritz Turbo can perform well in this test - and that some of the other products work 'a little' - but not that well. It is an extremely interesting experiment - and it certainly proves the question that started the thread: Bacteria in a bottle myth or fact.

When I was saying 'we dont really know what's going on in a cycle' - I meant that for example - I have used seachem stability many times - in clean sterile aquaria - and had no issues with fish loss/poor cycling, etc. I dont know why that it doesn't work here - but works in a natural aquarium - several others have posted similar results. Many people have stated that non-cooled bacteria in a bottle doesnt work as well as 'cooled bacteria' - yet Fritz 9 is non-cooled.

All I was trying to say is that the experiments bring up many more interesting questions yet to be answered - as @Dr. Reef has said. Thanks again to him for doing all the work.

Yes! Agree 100% with that statement! It is interesting. I agree. I am sold. I do not need further evidence to change my practice. Already done.

 

[lakai]

Why 2 ppm? if you added a tang for example - to a 100 gallon aquarium - nothing else - how long would it take to get to 2 ppm? If you added 2 clownfish to a 100 gallon tank - how long would it take to get to 2 ppm? What is the rationale for this number - except for dr. Tim saying - start at 2 ppn. Which is ridiculous *(unless he has data to suggest that level) - which isnt published. In the mean time - No one knows what they are doing with regards to cycling tanks IMHO

Think of it like anything living in your aquarium. Everything living is competing for food in order to survive and to grow. There are multiple strains of ammonia/nitrite oxidizing bacteria in bottled bac. Each one has a different set of environment conditions that allow for optimum growth such as temp, ph, oxygen and in this case concentration of Nh3.

Dose too much ammonia and you optimize growth of bacteria that works best in high nh3 eating up all the food that can hinder growth of the nh3 oxidizing bacteria we really want to dominate in our aquariums that thrive in low (under 6-7ppm) nh3 environments. Dose too little you will need to constantly monitor and feed.

Why 2ppm? Doesn't have to be but 2ppm< might prove difficult to test for in the case of the popular ammonia test kits which you need to match the color of your sample against the color on the card where in most cases there is a dramatic color shift at 2ppm which seems to be a compelling reason to start.

 

[Dr. Reef]

Just for clarification Seneye monitors only test for NH3 levels between 0 to 0.2 which would be in pH of 8.1 to 8.4 about 1-2 ppm TAN (NH3+NH4)

 

[MnFish1]

Think of it like anything living in your aquarium. Everything living is competing for food in order to survive and to grow. There are multiple strains of ammonia/nitrite oxidizing bacteria in bottled bac. Each one has a different set of environment conditions that allow for optimum growth such as temp, ph, oxygen and in this case concentration of Nh3.

Dose too much ammonia and you optimize growth of bacteria that works best in high nh3 eating up all the food that can hinder growth of the nh3 oxidizing bacteria we really want to dominate in our aquariums that thrive in low (under 6-7ppm) nh3 environments. Dose too little you will need to constantly monitor and feed.

Why 2ppm? Doesn't have to be but 2ppm< might prove difficult to test for in the case of the popular ammonia test kits which you need to match the color of your sample against the color on the card where in most cases there is a dramatic color shift at 2ppm which seems to be a compelling reason to start.

Just that I have never ever tested for ammonia with new tanks. (truth) - I have never had problems with deaths of fish. I apologize I dont believe that cycling a tank with a given amount of ammonia makes sense. I think - you add fish - you add a product to the tank - perhaps like @brandon429 says - its nitrifiers colonizing on their own - but it also debunks the 30 day myth that Dr Tim suggests. (apologies if thats not what he suggests). but people say dose x - and wait 30 days. I think thats bs. No offense to anyone that believes it...

 

[MnFish1]

Just for clarification Seneye monitors only test for NH3 levels between 0 to 0.2 which would be in pH of 8.1 to 8.4 about 1-2 ppm TAN (NH3+NH4)

But - isnt there a simple calculation between NH4 and NH3 levels depending on pH

 

[tankstudy]

What (we all know) - is that using Fritz Turbo at a 1 ppm and 2ppm and 8 ppm ammonia levels is that ammonia drops within 1-2 days to 0.

Was a bacterial density test ever conducted in this experiment?

 

[MnFish1]

Was a bacterial density test ever conducted in this experiment?

Right thats an issue - Turbo has a huge number of bacteria compared to the others - plus the instructions are to add a considerable amount. It is frankly not comparable. Not the fault of @Dr. Reef but the fault of the instructions on the bottles (which we discussed a couple pages ago)

 

[brandon429]

I have practical use for the knowledge of deposition times, example below
(Not the 2016 OP, the recent update question page two)
https://www.reef2reef.com/threads/c...onversion-seems-to-have-stalled.263968/page-2

All my cycling thread work is time based, not titration error of the day based. I can't trust anyone's reported numbers. One guy didn't shake the reagent enough; one lady is using the fw card not the marine one. One gentleman didn't fill to the five ml mark so his concentrate is off...One young person dosed prime and forgot to mention (a known nitrite test adulterant) there's always a reason that an APi test is ballparking if we're working in bulk reef threads, and time is the universal umpire.
Clearly DR had shown good ammonia APi lab technique no debate, but it's harder to rely on bulk public web post param levels as confirmed gold / need independent measures to not kill people's tanks when telling them to avoid hesitation due to a test kit that doesn't line up with known deposition time biology

Though his tests don't agree in that example link, he's cycled due to submersion time and boosters used...knowing how fast this occurs sooner than thirty days is vital for tuning no test cycle approaches.

We use, and won't be veering from 30 days anytime soon. Even if Dr Reefs tests show a three day turnaround on fritz, the ranging BB brands used, the ranging test kits used and unspoken cycle details, about thirty days is the soonest I'm comfortable calling cycles off submersion times. It'd be neat to see if day ten was a reliable cutoff but I can't test that in people's tanks like DR can test here, this side test is incredibly helpful information. It would be nice to know if even one slower performer can pull off the oxidation quick challenge at day ten, as a representation of the various bb brands people use when they post cycle times