Bacteria in bottle, busting myth, Seneye style.

[MnFish1]

so guys i came up on this test. how can this help us in our study.
https://www.hach.com/bart-test-for-nitrifying-bacteria-pk-7/product?id=7640250882

I looked this up in general - it looks like it tests bacteria in suspension - so theoretically you could add 'fritz' or whatever to a tank in the proper dilution and then immediately add the sample to the test vial - that would tell you the number of nitrifies in suspension at that time. With this warning:

1. 
   What about bacteria that are not active, aren't they important too?
   
   
   Bacteria can easily 'fall asleep' (become inactive) and it takes a while for these bacteria to wake up and become active again. In general the BART tests are not long enough to trigger these inactive bacteria to become active again. Because the cells are in the sleep mode then it is unlikely that their presence will have any impact on test and the results.

SO - maybe you add ammonia, add the fritz - (dont add substrate) - only a powerhead - and then measure after 12 and 24 hours (of course with 2 controls - 1 tank with fritz and no ammonia and one tank with neither)

 

[Dr. Reef]

Today I met with 3 manufacturers that I tested.
Fritz, Dr Tim, Brightwell
I had an excellent time at aquashella talking to many different manufacturers and vendors along with @brandon429 .
Thank you Brandon for meeting up with me. It was a pleasure.

Dr Tim shared a bunch of material that I will be posting soon.
@mikeyn at Fritz was very welcoming and shared number of details and behind the scene stuff.
I appreciate everyone for their time and information they shared with me.

 

[brandon429]

so happy to meet you guys!

 

[MnFish1]

Any updates?

 

[SciGuy2]

Yes, who won? Which product tastes best?

 

[Dr. Reef]

Ok guys i have a new take on this testing. I know we keep changing things but honestly i am changing things around as i am learning new things and from previous testing and the data collected from them.
Anyways, In my previous testing i used sterile tanks to test bacteria and from which i noticed that some bottles are True nitrifying bacteria and some are hetrotrophic.
Biggest problem with previous testing and i believe was a somewhat flaw, is that most all the manufacturers recommended adding fish then bacteria or vice versa. I used sterile tanks with pure ammonium chloride, Hetrotrophic bacteria do not work in sterile tank with only ammonia (atleast that what it showed in my testing) they require carbon source which fish produce.

So here is my new thinking on this next round of testing.

We dont know how much ammonia a fish produces every min every hour or every day so i have decided to run 5 tanks.
1 control and 4 bacteria tanks
Each tank will be 5 gal with 1 cup of sand and 500g of ceramic media.
I like to place a pair of clownfish in each tank and using 5 seneye monitors log how much ammonia each tank is producing.
For the 4 subject tanks i will dose ammonia just as the bottle suggests and in quantity it suggests.
Add fis then bacteria or add bacteria then fish etc.
I am sure after a day or 2 Control tank fish will die but i wont take them out and let ammonia spike and log all the data. while bacteria tanks will show a spike or not in ammonia and if those fish live or die etc i think would be more realistic and more colse to reality and more on manufacturers recommendation.
your thoughts?

 

[MnFish1]

Ok guys i have a new take on this testing. I know we keep changing things but honestly i am changing things around as i am learning new things and from previous testing and the data collected from them.
Anyways, In my previous testing i used sterile tanks to test bacteria and from which i noticed that some bottles are True nitrifying bacteria and some are hetrotrophic.
Biggest problem with previous testing and i believe was a somewhat flaw, is that most all the manufacturers recommended adding fish then bacteria or vice versa. I used sterile tanks with pure ammonium chloride, Hetrotrophic bacteria do not work in sterile tank with only ammonia (atleast that what it showed in my testing) they require carbon source which fish produce.

So here is my new thinking on this next round of testing.

We dont know how much ammonia a fish produces every min every hour or every day so i have decided to run 5 tanks.
1 control and 4 bacteria tanks
Each tank will be 5 gal with 1 cup of sand and 500g of ceramic media.
I like to place a pair of clownfish in each tank and using 5 seneye monitors log how much ammonia each tank is producing.
For the 4 subject tanks i will dose ammonia just as the bottle suggests and in quantity it suggests.
Add fis then bacteria or add bacteria then fish etc.
I am sure after a day or 2 Control tank fish will die but i wont take them out and let ammonia spike and log all the data. while bacteria tanks will show a spike or not in ammonia and if those fish live or die etc i think would be more realistic and more colse to reality and more on manufacturers recommendation.
your thoughts?

This should help:

https://www.researchgate.net/public...icity_to_false_clownfish_Amphiprion_ocellaris

As we discussed a while ago - and especially given this information in the article (where they recommend no ammonia higher than .57 ppm) and since you're using a Seneye -I think you could justify taking the fish out if the ammonia is higher than 0.7 or so (they used concentrations up I think 1.6 ppm) That said - I also think checking mortality is worthwhile. So either way - I thought you would be interested in the article. In another thread on another site - they calculated the amount t of ammonia produced by a clownfish -

I ran a Mr. Aqua 7.5 for QT with a pair of juvenile clowns, and had to do 5-gallon water changes daily to keep the ammonia levels down. That's with no live rock, so there was virtually no biofilter, but I was getting over 0.25 ppm in 24 hours. That would work out to about 0.05ml of 10% ammonia solution per day, if my use of online converters is accurate, plus any quantity that was successfully being converted to nitrite.

BTW - another key variable - is you should weigh out the food - to make sure that each tank is getting exactly the same amount of food - I think the food should also go into the control tank. (i.e. you should add one tank with no fish and just food added - to see how fast ammonia is building up just by feeding alone).

 

[hart24601]

While it messes with the controls I feel leaving fish in crosses an ethical line for me. Yes they are just fish but deliberately exposing to ammonia poisoning is where I would draw the line. Of course sometimes the information is valuable for aquaculture when done in a peer reviewed study, but I don't think it's a good idea here.

 

[MnFish1]

While it messes with the controls I feel leaving fish in crosses an ethical line for me. Yes they are just fish but deliberately exposing to ammonia poisoning is where I would draw the line.

Yet - isn't it true - that these products all recommend putting fish into the tank - with the bacteria? So - if the question is 'does bacteria in a bottle work' - you have to use fish. Fish are also exposed to ammonia poisoning every time they are shipped.

Having said that - Its likely - that the only tank that will have a problem is the control tank - (if the products work) - and we could pick a level at which the fish will be removed. it would still be a valid study - i.e. if the control fish needed to be removed after 1 day and bacteria XXX never had fish removed from that tank and in another tank YYY the fish needed to be removed after 6 days etc - it would still give meaningful information.

This study will also need to be repeated - so it might make sense the first time at least - to remove the fish if you're getting very accurate ammonia levels. JMHO

 

[Dr. Reef]

@hart24601 I agree with you 100% i dont want fish to die or to be subjected to ammonia but that leaves me with a problem,
without a control tank we wont know how much ammonia is being produced and if the other tanks are really working with bacteria added to them.
If anyone can come up with another way to test this, i can change the testing procedure.

 

[MnFish1]

@hart24601 I agree with you 100% i dont want fish to die or to be subjected to ammonia but that leaves me with a problem,
without a control tank we wont know how much ammonia is being produced and if the other tanks are really working with bacteria added to them.
If anyone can come up with another way to test this, i can change the testing procedure.

Check the study I posted on ammonia toxicity in ocellaris clowns... That post also has an estimate of ammonia production by a pair of clowns in a 7.5 gallon tank

 

[hart24601]

Check the study I posted on ammonia toxicity in ocellaris clowns... That post also has an estimate of ammonia production by a pair of clowns in a 7.5 gallon tank

Aye, what about it?

Ah are you suggesting to just add that ammonia then and not the fish?

 

[shred5]

@hart24601 I agree with you 100% i dont want fish to die or to be subjected to ammonia but that leaves me with a problem,
without a control tank we wont know how much ammonia is being produced and if the other tanks are really working with bacteria added to them.
If anyone can come up with another way to test this, i can change the testing procedure.

You said they need a carbon source and that comes from fish? Why not just add the carbon to the water?
Bacteria require nitrogen, carbon and phosphorus correct? All three should not be to hard to add..

Adding fish is going to turn this thread into a mess with people against doing that!.

 

[hart24601]

@hart24601 I agree with you 100% i dont want fish to die or to be subjected to ammonia but that leaves me with a problem,
without a control tank we wont know how much ammonia is being produced and if the other tanks are really working with bacteria added to them.
If anyone can come up with another way to test this, i can change the testing procedure.

Yes it does mess up the controls, however I feel we should keep in mind the purpose and context of these experiments. Hobby level to show if bottled bacteria work. Is this really needed and valuable to the point of killing fish? As I said this isn't going to be peer reviwed aquaculture study that have ethics committee. I can see a lot of people not liking this approach.

 

[Dr. Reef]

You said they need a carbon source and that comes from fish? Why not just add the carbon to the water?
Bacteria require nitrogen, carbon and phosphorus correct? All three should not be to hard to add..

Correct and that i have already tested in my other thread and study but most manufactorers stalled in sterile tanks and they said it stalled because i didnt add fish to the system like bottle suggests. so to be fair to the companies i wanted to design a test that would be more realistic and close to what bottle suggests.

 

[MnFish1]

Aye, what about it?

Ah are you suggesting to just add that ammonia then and not the fish?

No the study suggests the 'toxic level' of ammonia for clownfish. If the level in a tank got up to (pick your level) - the fish would be removed. Remember in this study - there will be continuous ammonia monitoring.

 

[MnFish1]

Correct and that i have already tested in my other thread and study but most manufactorers stalled in sterile tanks and they said it stalled because i didnt add fish to the system like bottle suggests. so to be fair to the companies i wanted to design a test that would be more realistic and close to what bottle suggests.

Dr. Tim's worked at 2 and 4 PPM didnt it (though slower than fritz) with just ammonia? (It must have - since Dr. Tims recommends adding only ammonia)

 

[hart24601]

No the study suggests the 'toxic level' of ammonia for clownfish. If the level in a tank got up to (pick your level) - the fish would be removed. Remember in this study - there will be continuous ammonia monitoring.

That seems to be a reasonable approach to me.

 

[Dr. Reef]

The only other way around is to put a pair of clownfish in a sterile tank (uncycled) and using Seneye monitor log ammonia production and pull the fish out after few hours or at/about NH3 reaches 0.2. As we know 0.2 and up ammonia will start to damage organs.
Once i have the time and ammonia produced in that time from i can calculate ammonia production per hour and then simply using a doser add that much every hour.

 

[MnFish1]

Yes it does mess up the controls, however I feel we should keep in mind the purpose and context of these experiments. Hobby level to show if bottled bacteria work. Is this really needed and valuable to the point of killing fish? As I said this isn't going to be peer reviwed aquaculture study that have ethics committee. I can see a lot of people not liking this approach.

The instructions for these products suggest adding the bacteria - and fish at the same time so - how is this unethical. The control tank could be an issue - and there is a suggestion on 'what to do' without harming the fish.

A lot of people dont believe in using fish with cycling or bacteria at all. So - IMHO this is the only way to get the answer. My guess is that it will prove many thoughts that are 'common sense' wrong concerning fish and bottled bacteria.