Bacteria in a bottle, Myth or Fact

[SDReefer]

Yes the 2 control tanks will be just simply tanks put together identically as all 4 but without the bacteria added to them. They are control theoretically they should take the longest to cycle vs tanks with bacteria added should cycle faster. if the claims from companies are true. boiling the bottle bacteria will kill the bacteria and will be no use in any way. am i missing the point? elaborate please.

I'm assuming that bottled bacteria isn't purely bacteria - it also has other compounds, trace elements, etc in the liquid itself. To make all things equal, adding an 'inactive' form of the bottled bacteria would make it purely control (the only difference is whether the bacteria is alive or dead).

I'm going to tag @Randy Holmes-Farley so that we can get input from someone who's done serious research.

 

[Dr. Reef]

Looking forward to see the result from all tests regarding Ammonia as many tanks, including my nano have shown 0.25 readings from day of set up. I used established live rock with a splash of old tank water, then new synthetic saltwater and ro washed new reeflowers sand.

Is it the bacteria in the bottle that holds the key, or the test kits not being accurate enough at hobbyist level?

@brandon429 you got me challenging this madness *grin*

A.

initially i started off with the assumption that such products were snake oil but within few posts i did realize that there is truth to some of these products. now the focus has shifted from proving if they work or not to how effective these products are. like i mentioned in previous post some products are live some spores and some blends they all should have differences in the way they cycle a tank and in time how long they take.
So now its all about length of processing ammonia and testing ammonia test kits against each other for accuracy as well.

 

[Dr. Reef]

I'm assuming that bottled bacteria isn't purely bacteria - it also has other compounds, trace elements, etc in the liquid itself. To make all things equal, adding an 'inactive' form of the bottled bacteria would make it purely control (the only difference is whether the bacteria is alive or dead).

I'm going to tag @Randy Holmes-Farley so that we can get input from someone who's done serious research.

good point.
I am not going to mention the name but one of the big name brand company has promised to provide some helpful suggestions regarding this experiment and i am awaiting to see what they have to say. maybe they can guide me and us to a perfect experiment without too many variables to alter results. i am open to all suggestions to make this as accurate as it can be,

 

[Dr. Reef]

People that think that "how can bacteria stay in a bottle, they need to eat!" lack a basic understanding of the biology of bacteria. Bacteria can stay in stasis for thousands of years, yes thousands of years. It is all about how the bacteria is processed and packaged. It is a worthwhile test for sure but you didn't get some of the best bacteria available.. biodigest from probidio. There oxygen free packaging ensures that no bacteria are lost. I just cycled my 40 gallon breeder QT tank in two days and I added 5ppm of dr tims ammonia.

its added to the list and email sent to manufacturer.

 

[Dr. Reef]

the problem with this is, i am guessing we will see some pretty good results. however the rest of us might not get the freshest bottle. would do one fresh and one from a random store. be interesting to what the results are. i know more $$ but if the companies are offering the product for free then might be worth it?

I am really looking forward to this thread!

I like that idea. You're absolutely right getting a fresh bottle from the manufacturer will definitely give us good results but I like to compare it to a random bottle purchased online or locally to see what the differences are let's see what others have to say about it

 

[hart24601]

Some interesting links on fritz site.

http://fritzzyme.com/files/Identification of nitrifying bacteria contained in a commercial i-1.pdf

 

[Diznaster]

Have you considered testing "Purple Helix"? It is sold as coralline spores with nitrifying bacteria (heterotrophic, chemolithotrophic, autotrophic). It is bottled and shipped immediately when ordered and says to use within 14 days. I started a tank a couple months ago with reef saver rock and have not added anything else that would have contributed coralline and I can confirm it works for that. My rocks are quickly turning green and purple in a good way. A friend at work also used this with similar results. I cycled my tank with Red Sea Reef Mature, it took 3 weeks, but now at 8 weeks I actually have NO3/PO4 too low. I have a couple 4" thick MarinePure ceramics in the sump, and I am understocked for fish and adding more gradually. No corals yet. So I can't really say what the efficacy of the bacteria in Purple Helix is, but I'd bet it is fresh and good since their coralline spores are legit.

 

[bluewill]

I think the test is valid .Every positive post I've read about personal experiences of great performance surely could be valid ,and it makes perfect sense that this should work .I think Dr. Reef is questioning whether the bacteria is alive when purchased (please correct me if I'm wrong ). I haven't seen any comparisons of results using the same parameters without the addition of a bacteria supplement. I see many members using "survival of fish" as the proof of the benefit of these products.Other members show actual details of ammonia reduction rates, but none with comparisons to a non supplemented equal.I definitely agree to remove the "dry rock" , it adds to many variables to the test .

 

[Diznaster]

this will take a long time to test each product but if i am not mistaken we dont need to do control tanks each time?

Probably not a bad idea to run a control tank during every round. The possible problem I see is if the environmental conditions change it could effect the results and you would not know it. Something unknown and not measured could skew the results. With a control run for every round you could look at the standard deviation of all the control runs and get a decent understanding of repeatability/uncertainty. I'd be happy to expand on the topic or try to help with that even though I really don't enjoy calculating uncertainty of measurement processes. I do it for dimensional metrology but the same concepts would apply but are going to be somewhat coarse and easier here. We certainly are not going to measure the repeatability of drips from the reagents in the test kits, etc...

Edit: Actually after thinking about this more, if you are using multiple test kits and do multiple control tanks it should be possible to get a general idea of the repeatability of each test kit and the repeatability of each control tank. If you can log the data into Excel I can do the math.

 

[SEMA]

I've used API, brightwell, and instant ocean bacterial additives in commercial applications for over 12yrs with finominal results. Brightwell has been my go to product from a market standpoint. I've been keeping my livestock loss under 1% with very heavy stocking density in the first 30 days..

 

[Dr. Reef]

Guys i just got done talking to Scott with Ecological Labs manufacturers of Microbe Lift products. It was very nice to talk to him and he provided very good information about his products and background of his company. He recommended the product NiteOut 2 for our experiment. Its is a live bacteria with shelf life of 14-16 months. They also committed to provide their product when time comes for experiment so i really like to thank Scott and Ecological Labs to reach out to us and help make this experiment possible.

 

[Dr. Reef]

So far i have commitments from: (in order they came in)

1. Fritz
2. Dr. Tim
3. Microbe Life (Niteout 2)

I recommend all hobbyist to give these manufacturers a great hand of applause and thank you for their commitments and their work in our industry and helping the environment. They all sound very confident about their products and have research and years to prove it. I myself when i started my hobby 25-30 yrs ago things were very different and i was very skeptical of such products i guess you can call me old school. This is a great learning experience for myself and i am sure the data points are going to be more interesting for all of us.
I like to add that this experiment is not to put one product down or over the other. simply want to see which product works in what length of time.

BWT most of these companies offer tours of their facilities if someone is interested.

 

[Mohammad D. ALMUTAIRI]

1st of all a big thank you to you Dr. Reef for trying this testing and giving the time and effort to see it through

2nd I'd like to give my point view to the testing method used here with 4 tanks

I'd go with
1st tank being setup the way you say and be left alone to see if it cycle as normal with time
2nd tank being setup as you said but instead of being left we add pure ammonia to it, without adding no bacteria in the bottle product to it, to see if it cycle faster than normal tank "or the old method" we used to cycle with
3rd thank being adding the bacteria into it without adding ammonia, but adding fish food "ghost feeding" or a dead shrimp. simply the old method again but with bacteria.
4th tank is where we add both pure ammonia and bacteria to the tank and see how much it gonna take to cycle

this is my take on the subject, being right or wrong, I don't know, but this is my 2cent on it

once again, thank you for doing so, it means a lot to a hobbyist like me, who want to learn a new thing everyday.

good luck and god bless

 

[brandon429]

Your thread here, after these tanks are cycled and the readings are complete can be used to branch off into powerful statements about what bacteria do and do not tolerate, which controls about 80% of the allowable and allowed actions in this hobby.

Recall in 1998 same reefs: any water change over ten pct/dangerous / not

So your cycled tanks need to be ran even still for a while, you have fallow testing (do bac require our feed) and you have hyposalinity testing here to validate

https://www.reef2reef.com/threads/cycle-during-hyposalinity.382857/#post-4821161

And there's upper and lower tank temp testing, basically you can discover what nature tolerates, after the retail portions are weighed and assessed.

This is fifty times what you perhaps wanted to do/awr = let someone else take off on the ideas. Let's stress test the final cycled tanks like myth busters does see what it takes to effect the myth

 

[Dr. Reef]

Your thread here, after these tanks are cycled and the readings are complete can be used to branch off into powerful statements about what bacteria do and do not tolerate, which controls about 80% of the allowable and allowed actions in this hobby.

Recall in 1998 same reefs: any water change over ten pct/dangerous / not

So your cycled tanks need to be ran even still for a while, you have fallow testing (do bac require our feed) and you have hyposalinity testing here to validate

https://www.reef2reef.com/threads/cycle-during-hyposalinity.382857/#post-4821161

And there's upper and lower tank temp testing, basically you can discover what nature tolerates, after the retail portions are weighed and assessed.

This is fifty times what you perhaps wanted to do/awr = let someone else take off on the ideas. Let's stress test the final cycled tanks like myth busters does see what it takes to effect the myth

Awesome idea. Thats what i will do after thr product comparisons.
Lets stress test them and break them. See what it takes to kill it. Temp salinity hyposalinity copper etc

 

[brandon429]

excellence.

and the grand one= emersion time tolerance. My own pico reef which is 12 yrs old gets drained routinely, as in about a thousand times, for half an hour to simulate tidal flows and for personal amusement.

plus from 2010 there's simon garratts intertidal reef which is drained for six hours a day, every day, with huge acropora in tow.

my half hour which is most I can stand to do lol and his six hours = established nitrifiers are darn near bulletproof regarding emersion.

I also have reports in my microbiology thread about live rock sitting in a dark closet for two months and still passing an oxidation test within 24 hours of submersion back in a test vat.




so online sources state that they don't tolerate drying, but that doesn't mean draining a tank with ironclad biofilm insulations counts as drying, nope. it means pure strain liquid suspensions of the bacteria which are not in biofilm (since they can be poured out of a bottle and not remain stuck to its sides) can be dried easily.

what it takes to kill surface area cycled nitrifiers is just astonishing. they tolerate massive amnts of peroxide and all kinds of weird things.

 

[B-ridge]

I’m just curious to know what these bacteria in a bottle are feeding on once inside the bottle?

 

[Dr. Reef]

From what ive been told they are packaged with amino acids to keep them alive.

 

[Ferrell]

I’ve cycled a 75 display and a 10 QT with dr tims and was stress free for the fish both times. Had to do an h2o change on the qt because of the number of fish and high NO3=10 but no huge ammonia spike at all

 

[DrTim]

Hello Everyone:

DrTim from DrTim's Aquatics and the developer of DrTim's One and Only. I thought I would try to answer some of the questions about how bacteria can stay alive in a bottle. I can only speak for my product and what we did at Marineland with BioSpira which I also developed. I have been studying and growing nitrifiers for over 20 years for reference so have a lot of experience with these bugs.

First and most obvious bacteria, including nitrifying bacteria, are not human. They do not have lungs or much of anything else humans or other multi-cellular organisms have in terms of needs to survive. So first, they do not need oxygen to survive. They need oxygen to convert ammonia to nitrite or nitrite to nitrate but that is to gain energy which to a bacterium is different than surviving. So the liquid in the bottle does not need a constant supply of oxygen unless one wants the bacteria to be dividing in the bottle. Thus if you don't add ammonia to the bottle there is not need to supplement with oxygen and the bacteria do quite well.

So if don't they feed (no ammonia or nitrite in the bottle) and don't require oxygen how to they survive. The answer is that all bacteria have cellular mechanisms to adapt and survive when conditions are not good for them. Many bacteria form spores. Nitrifiers do not form spores but produce a sugar inside that cell that helps them survive in poor conditions.

The other thing about bacteria is that they are hard to kill. Unless you break the cell open or disrupt the DNA/RNA or poison the cell (and some other things) the bacterium survive in a viable state. That is why they have been around for so long. Further, nitrifying bacteria live in a biofilm that acts as additional protection.

So how can they survive? Think of the nitrifier cell as a rechargeable battery. It's bottled at 100% energy level and since the organism does not have any ammonia or nitrite to feed on it does not have to spend any energy reproducing. Also since the conditions are not optimal for reproducing it goes into a resting stage and uses very little energy from its energy center waiting for conditions to improve. In this state they can last a long time in a bottle and work once poured into your tank. And the last key things is that the nitrifiers have to be the right species but that another story!

While many people think heat kills nitrifiers that is not true unless you get the liquid really hot what kills them is freezing. If the liquid is frozen solid for a period of time the ice crystals that form puncture the cell wall killing the bacteria.

So keep the nitrifying bacteria viable in a bottle you don't need secret sauce, you don't need to add amino acids or ammonia or nitrite. You need the have the right bacteria in the bottle to start (the key difference among products) and you need to keep the bottle from freezing.

I hope this explains a few things about nitrifiers - DrTim