My speculation: Vibrant has some fluconazole in it...

[nereefpat]

Here is one of the posts - there was a lot of discussion about this. There are a couple other experiments that were also done using broth - and the broth samples were sent to aquabiomics - and the bacteria genus that were found were posted (i.e. the report from Aquabiomics).

My speculation: Vibrant has some fluconazole in it...

So... Why does everyone care so much? It works wonders in my tank... I hesitate to even post on the subject because it seems like vibrant is a product people love to hate. I started using vibrant and my tanks have literally never been cleaner. I am very interested in the rise of microbial...

www.reef2reef.com

The Aquabiomics sample was a mixture of 7 different 'additives,' not a sample of Vibrant, it looks like. That doesn't tell us much about bacteria in Vibrant.

As for the culturing, that is interesting. To me, the curve for optical density does not track as a bacterial culture. A couple of the other additives don't look so hot either. It isn't sterile (or at least isn't kept sterile during the test), which isn't surprising. Also there aren't enough cells there, which aligns with the centrifuge pictures. This isn't a bacterial culture. 3 1/2 days before log growth? Come on.

 

[MnFish1]

The Aquabiomics sample was a mixture of 7 different 'additives,' not a sample of Vibrant, it looks like. That doesn't tell us much about bacteria in Vibrant.

True - it was somewhat unclear. Now I have to re-read it lol
EDIT - you are correct - it was just 'a pooled sample'

As for the culturing, that is interesting. To me, the curve for optical density does not track as a bacterial culture. A couple of the other additives don't look so hot either. It isn't sterile (or at least isn't kept sterile during the test), which isn't surprising. Also there aren't enough cells there, which aligns with the centrifuge pictures. This isn't a bacterial culture. 3 1/2 days before log growth? Come on.

UWC also posted a bacterial curve - on the thread - I'll try to find that. But - I think EDIT - THE ARTICLE BELOW answers your concerns about optical density. It certainly depends on whether you're starting out with 'bacteria' or with 'spores'. If you just go to the conclusions part of the article - it explains why the OD method - in this situation - may not be 'the best' (or even good) - at estimating bacterial number. 1) the OD depends not just on number but also 'type' of bacteria.
2) the OD depends on the bacteria in the suspension being in the same 'growth phase'.
3) The growth medium may favor one over another bacteria. I would imagine - that these products 'if they work' - attack algae, etc on solid surfaces as compared to 'seawater nutrients'. Thus - the comparison to seawater may not be accurate either.

http://microbiologynetwork.com/measurement-of-cell-concentration-in-suspension-by-optical-density.asp

 

[nereefpat]

It certainly depends on whether you're starting out with 'bacteria' or with 'spores'. If you just go to the conclusions part of the article - it explains why the OD method - in this situation - may not be 'the best' (or even good) - at estimating bacterial number.

OD600 is not for exact CFUs/mL, I agree. The best method for that would be to plate it. The exact number isn't really that important to me here. There is so little, that I don't consider it a bacterial additive. It's certainly not a culture. There is so little that you can't see cells in a centrifuged sample, and so little that when cultured it takes several days to see any growth.

1) the OD depends not just on number but also 'type' of bacteria.

Yes, but we're not looking for exact numbers here. I know we strive for quantitative when we can. But if there are 2 cfus per 100mLs or 20, that's still 'too few to count.'

2) the OD depends on the bacteria in the suspension being in the same 'growth phase'.

When making stocks of bacteria (or yeast) used to inoculate a flask, fermenter, or whatever, if you make the stocks from a culture during the log-phase growth, they will take off faster in the new system. If that's what you're saying, I have seen that.

3) The growth medium may favor one over another bacteria.

Yes. But the Vibrant eventually had the expected curve, after enough days. I just think there isn't enough there.

For the company to call this stuff "95% cultured bacteria blend" is laughable. The directions call for 1 mL per 10 gallons of water. For a 100 gallon aquarium, you would dose 10mLs. Look at the 10 mL centrifuge sample. There aren't enough cells to inoculate anything.

 

[MnFish1]

OD600 is not for exact CFUs/mL, I agree. The best method for that would be to plate it. The exact number isn't really that important to me here. There is so little, that I don't consider it a bacterial additive. It's certainly not a culture. There is so little that you can't see cells in a centrifuged sample, and so little that when cultured it takes several days to see any growth.

Yes, but we're not looking for exact numbers here. I know we strive for quantitative when we can. But if there are 2 cfus per 100mLs or 20, that's still 'too few to count.'

When making stocks of bacteria (or yeast) used to inoculate a flask, fermenter, or whatever, if you make the stocks from a culture during the log-phase growth, they will take off faster in the new system. If that's what you're saying, I have seen that.

Yes. But the Vibrant eventually had the expected curve, after enough days. I just think there isn't enough there.

For the company to call this stuff "95% cultured bacteria blend" is laughable. The directions call for 1 mL per 10 gallons of water. For a 100 gallon aquarium, you would dose 10mLs. Look at the 10 mL centrifuge sample. There aren't enough cells to inoculate anything.

I am hoping the company will give out more Information as they did in the beginning of the theead

 

[taricha]

I had wanted to revisit my bacterial growth results posted around page 3 of this thread to see if I did it wrong or if in fact bacteria can survive in the quat-like chemical detected in Vibrant and the known quat in AlgaeFix. The short version is that random contamination in the bottles can explain everything I've seen so far just fine.
This is instant ocean at 1.026 enriched with LB broth + glucose (same media that I cultured up products on in page 3.) Picture after 7 days.

I added 1 drop (0.05mL) of each of the following
1 & 2 duplicates - Distilled water (stayed clear for 7+ days)
3 & 4 duplicates - Vibrant run through an 0.22um syringe filter (stayed clear for 7+ days)
5 & 6 duplicates - Vibrant the same, except syringe filter removed (cloudy by 57hr)
7 AlgaeFix (cloudy by 45hr)
8 MicroBacter7 (cloudy by 48 hours)
9 Aquarium water (cloudy by 18 hrs)

Takeaways:
1) I'm not crazy nor was the culture-up necessarily flawed. Vibrant did contain viable bacteria, which could be excluded by a syringe filter.
2) AlgaeFix also cultured up bacteria just the same, so bacteria can survive in the polyquat in question.
3) This kind of culture up does not distinguish between bona fide spore containing products (MB7) and irrelevant contamination in non bacterial products (AlgaeFix). AlgaeFix clouded up faster in fact.
4) therefore nothing can be concluded from my previous culture-up of vibrant (and the other products) except that they are not sterile. Contamination can exist in vibrant just as easily as in algaefix.
5) point 4 about random contaminants is further evidenced by the aquabiomics report in post 58 that showed some of the most prevalent bacteria from my combined cultures to be strains that no one would want and have no business in bottled products. Aquabiomics told me that by email, many of the strains found are not ones that would be added intentionally (most prevalent was a genus associated with fish disease).

 

[MnFish1]

I had wanted to revisit my bacterial growth results posted around page 3 of this thread to see if I did it wrong or if in fact bacteria can survive in the quat-like chemical detected in Vibrant and the known quat in AlgaeFix. The short version is that random contamination in the bottles can explain everything I've seen so far just fine.
This is instant ocean at 1.026 enriched with LB broth + glucose (same media that I cultured up products on in page 3.) Picture after 7 days.

I added 1 drop (0.05mL) of each of the following
1 & 2 duplicates - Distilled water (stayed clear for 7+ days)
3 & 4 duplicates - Vibrant run through an 0.22um syringe filter (stayed clear for 7+ days)
5 & 6 duplicates - Vibrant the same, except syringe filter removed (cloudy by 57hr)
7 AlgaeFix (cloudy by 45hr)
8 MicroBacter7 (cloudy by 48 hours)
9 Aquarium water (cloudy by 18 hrs)

Takeaways:
1) I'm not crazy nor was the culture-up necessarily flawed. Vibrant did contain viable bacteria, which could be excluded by a syringe filter.
2) AlgaeFix also cultured up bacteria just the same, so bacteria can survive in the polyquat in question.
3) This kind of culture up does not distinguish between bona fide spore containing products (MB7) and irrelevant contamination in non bacterial products (AlgaeFix). AlgaeFix clouded up faster in fact.
4) therefore nothing can be concluded from my previous culture-up of vibrant (and the other products) except that they are not sterile. Contamination can exist in vibrant just as easily as in algaefix.
5) point 4 about random contaminants is further evidenced by the aquabiomics report in post 58 that showed some of the most prevalent bacteria from my combined cultures to be strains that no one would want and have no business in bottled products. Aquabiomics told me that by email, many of the strains found are not ones that would be added intentionally (most prevalent was a genus associated with fish disease).

Agree you're not crazy. . This is a great experiment!

1. Not all bacteria are excluded by a .22 micron filter. It would have been nice to see the same results with the other bacteria products including algaefix. But - it seems that the 'bacteria' in Vibrant is excluded by a .22 micron filter. - What would have been extremely interesting - is if the same thing happened with algaefix? The types of bacteria NOT excluded by a 22 micron filter - are multiple - but - not common to use (I would think) in this situation
2. I believe that the products - no matter what they are - are not completely 'the desired bacteria'. Again here - a quantitation of those bacteria is important. I.e. - How sensitive is aquabiomics.
3. This makes me curious. I have no idea - whether spores - have the same DNA fingerprint as the whole organism. Thats a question for @AquaBiomics

See the next post for more information (if you're still not geeked out with microbiology)

 

[MnFish1]

OD600 is not for exact CFUs/mL, I agree. The best method for that would be to plate it. The exact number isn't really that important to me here. There is so little, that I don't consider it a bacterial additive. It's certainly not a culture. There is so little that you can't see cells in a centrifuged sample, and so little that when cultured it takes several days to see any growth.

Yes, but we're not looking for exact numbers here. I know we strive for quantitative when we can. But if there are 2 cfus per 100mLs or 20, that's still 'too few to count.'

When making stocks of bacteria (or yeast) used to inoculate a flask, fermenter, or whatever, if you make the stocks from a culture during the log-phase growth, they will take off faster in the new system. If that's what you're saying, I have seen that.

Yes. But the Vibrant eventually had the expected curve, after enough days. I just think there isn't enough there.

For the company to call this stuff "95% cultured bacteria blend" is laughable. The directions call for 1 mL per 10 gallons of water. For a 100 gallon aquarium, you would dose 10mLs. Look at the 10 mL centrifuge sample. There aren't enough cells to inoculate anything.

Sorry I was a little lazy - I quoted the whole thing. Im not going to argue (right or wrong) - the 95% xxxxxx. I do not know the rules for active ingredients, etc.

Think about this perhaps- remember - 'In reefing - nothing happens fast (I don't agree with this) - and 'stability is key''

If I were to manufacture a product - I would want that product to ramp up slowly - Otherwise - lets be real - its true - there would be bacterial blooms in all the tanks - stability affected, ETC ETC ETC. I think you make good points. Like I said - Nothing I have seen (and I and probably no one else here) - is expert enough in all of these testing procedures to make an honest comment - since they cover so many disciplines,

AFAik vibrant - does not claim to kill algae on day 1. or 2 - which is good. (To me) -

@taricha - thanks for repeating your bacterial experiments!

 

[MnFish1]

Agree you're not crazy. . This is a great experiment!

1. Not all bacteria are excluded by a .22 micron filter. It would have been nice to see the same results with the other bacteria products including algaefix. But - it seems that the 'bacteria' in Vibrant is excluded by a .22 micron filter. - What would have been extremely interesting - is if the same thing happened with algaefix? The types of bacteria NOT excluded by a 22 micron filter - are multiple - but - not common to use (I would think) in this situation
2. I believe that the products - no matter what they are - are not completely 'the desired bacteria'. Again here - a quantitation of those bacteria is important. I.e. - How sensitive is aquabiomics.
3. This makes me curious. I have no idea - whether spores - have the same DNA fingerprint as the whole organism. Thats a question for @AquaBiomics

See the next post for more information (if you're still not geeked out with microbiology)

I had wanted to revisit my bacterial growth results posted around page 3 of this thread to see if I did it wrong or if in fact bacteria can survive in the quat-like chemical detected in Vibrant and the known quat in AlgaeFix. The short version is that random contamination in the bottles can explain everything I've seen so far just fine.
This is instant ocean at 1.026 enriched with LB broth + glucose (same media that I cultured up products on in page 3.) Picture after 7 days.

I added 1 drop (0.05mL) of each of the following
1 & 2 duplicates - Distilled water (stayed clear for 7+ days)
3 & 4 duplicates - Vibrant run through an 0.22um syringe filter (stayed clear for 7+ days)
5 & 6 duplicates - Vibrant the same, except syringe filter removed (cloudy by 57hr)
7 AlgaeFix (cloudy by 45hr)
8 MicroBacter7 (cloudy by 48 hours)
9 Aquarium water (cloudy by 18 hrs)

Takeaways:
1) I'm not crazy nor was the culture-up necessarily flawed. Vibrant did contain viable bacteria, which could be excluded by a syringe filter.
2) AlgaeFix also cultured up bacteria just the same, so bacteria can survive in the polyquat in question.
3) This kind of culture up does not distinguish between bona fide spore containing products (MB7) and irrelevant contamination in non bacterial products (AlgaeFix). AlgaeFix clouded up faster in fact.
4) therefore nothing can be concluded from my previous culture-up of vibrant (and the other products) except that they are not sterile. Contamination can exist in vibrant just as easily as in algaefix.
5) point 4 about random contaminants is further evidenced by the aquabiomics report in post 58 that showed some of the most prevalent bacteria from my combined cultures to be strains that no one would want and have no business in bottled products. Aquabiomics told me that by email, many of the strains found are not ones that would be added intentionally (most prevalent was a genus associated with fish disease).

Curious - how do you know its not bacteria in the salt>?

 

[Ghost25]

Ultimately I think only LC/MS will resolve this to the satisfaction of the doubters. In the meantime it has been established that AlgaeFix and Vibrant behave very similarly. Falsification is the core of the scientific method.

Hypothesis: there is no significant difference between AlgaeFix and Vibrant.

So to those who doubt the above hypothesis: demonstrate an experiment showing a significant difference between the two. I really don't have a dog in the fight, but what I don't like, and what is an unfortunate consequence of human psychology is the tendency to latch on to an idea and seek to validate it at all costs, discarding any evidence that contracts their idea in order to rescue it.

Seriously, I don't care if Vibrant has QAC or not, but given the evidence that AlgaeFix and Vibrant are very similar, I am inclined to think that it does. Demonstrate the two products are significantly different and you will be on the way to falsifying that claim.

 

[MnFish1]

Ultimately I think only LC/MS will resolve this to the satisfaction of the doubters. In the meantime it has been established that AlgaeFix and Vibrant behave very similarly. Falsification is the core of the scientific method.

Hypothesis: there is no significant difference between AlgaeFix and Vibrant.

So to those who doubt the above hypothesis: demonstrate an experiment showing a significant difference between the two. I really don't have a dog in the fight, but what I don't like, and what is an unfortunate consequence of human psychology is the tendency to latch on to an idea and seek to validate it at all costs, discarding any evidence that contracts their idea in order to rescue it.

Seriously, I don't care if Vibrant has QAC or not, but given the evidence that AlgaeFix and Vibrant are very similar, I am inclined to think that it does. Demonstrate the two products are significantly different and you will be on the way to falsifying that claim.

But - in fact, there are significant differences. I'm sorry if you haven't been following along - I'm not inclined to list all of them 'again'. However - you are indeed correct - in the sense that the differences - like the similarities - are more correlation - than causative. Lets start here:

1. There is bacteria in vibrant - is it more or less than algaefix (we don't know that experiment was not done)
2. The pH are the same? Is the buffering capability the same ( a known interference point for bromphenol blue testing of Quats) - IDK
3. A precipitate formed during one of the methods - Which - IMHO invalidates that method for this topic - but it was continued - with different parameters. For anyone who cares - there are multiple different methods for measuring Quat's - in milk, on clothing, ETC ETC ETC. They all differ.
4. The original method for measuring QUATS - was used in a study for residual Quat's - that were on table tops after wiping and drying. I.e. very small amounts. If I remember correctly , UWC has already said there are small amounts of Quat's in Vibrant - So - the test is positive. There is no evidence - and IMHO - there cannot be - that the blue color visible to the eye - is x shades darker with vibrant than algaefix.

So - In the end I agree with you completely - its the companies turn to give the proof in the pudding. The amateur scientists (myself included) - want to know - whats going on here?

 

[taricha]

Curious - how do you know its not bacteria in the salt>?

...then it would have grown bacteria in the distilled and filtered vibrant samples. All were added to the same enriched sterilized sw media.

I know that you tested the pH of Vibrant and Algaefix - did you happen to test the alkalinity?

At the amounts added in the test, (<0.5uL product per mL) maybe something in the hundredths of a dKH. 20x that concentration came out barely detectable, at like 0.5dKH or so.

Demonstrate the two products are significantly different and you will be on the way to falsifying that claim.

this is my question right now as well.
I have not come up with any test that distinguishes them.

Here's another fun coincidence.
Dried residue. Each sat overnight on a coffee warmer.

3.00mL AlgaeFix = 0.177g
3.00mL Vibrant = 0.182g
(3.00mL water for comparison and uncertainty scale = 0.003g)

 

[MnFish1]

...then it would have grown bacteria in the distilled and filtered vibrant samples. All were added to the same enriched sterilized sw media.


At the amounts added in the test, (<0.5uL product per mL) maybe something in the hundredths of a dKH. 20x that concentration came out barely detectable, at like 0.5dKH or so.


this is my question right now as well.
I have not come up with any test that distinguishes them.

Here's another fun coincidence.
Dried residue. Each sat overnight on a coffee warmer.

3.00mL AlgaeFix = 0.177g
3.00mL Vibrant = 0.182g
(3.00mL water for comparison and uncertainty scale = 0.003g)

Maybe then the postulate should be that algaefix also contains bacteria??? IDK. Since (which I thought you had done before - maybe that was your Prime experiment) - you compared bacteria - in both products.

 

[ScottB]

This is certainly true. BUT - one thing you said is a common misconception. There are plenty of 'harmful' bacteria. Vibrio, for example among many others. I'm almost just as paranoid about adding a bacterial product as I am a known chemical - with a known concentration. Why? Because bacteria can grow much differently in 1 tank (with one set of nutrients) as compared to another tank with a different set of nutrients, etc, etc. At least with a 'chemical' its possible to remove it with water changes.

Which vendor is selling vibrio as a bacteria? Enquiring minds want to know. Did Aquabiomics turn up with vibrio in their tests of the "popular" bacteria bottles? Maybe I missed that. Or I don't understand what you are suggesting.

We don't know what is in any of them. I quit using any of them. Live rock works just fine.

 

[MnFish1]

Which vendor is selling vibrio as a bacteria? Enquiring minds want to know. Did Aquabiomics turn up with vibrio in their tests of the "popular" bacteria bottles? Maybe I missed that. Or I don't understand what you are suggesting.

We don't know what is in any of them. I quit using any of them. Live rock works just fine.

Read what I said in context - I'm not going to repeat it and go through a bunch of other stuff. My statement stands in response to what I said

A hint - there are a lot of harmful bacteria - that was one example - You(someone) said - I'm nervous about adding chemicals - but - bacteria are safe - paraphrased. Go read what you or whoever it was that wrote it It was not an endorsement of vibrio - I could have picked numerous bacteria - that are a lot more harmful than Algaefix

 

[taricha]

Maybe then the postulate should be that algaefix also contains bacteria???

Right. That was my conclusion.

2) AlgaeFix also cultured up bacteria just the same, so bacteria can survive in the polyquat in question.
3) This kind of culture up does not distinguish between bona fide spore containing products (MB7) and irrelevant contamination in non bacterial products (AlgaeFix). AlgaeFix clouded up faster in fact.
4) therefore nothing can be concluded from my previous culture-up of vibrant (and the other products) except that they are not sterile. Contamination can exist in vibrant just as easily as in algaefix.

 

[MnFish1]

Right. That was my conclusion.

No I meant functional bacteria - not bacterial contaminants. -perhaps you can look at which bacteria survive in 4.5 % QUAT - and I'm sure there are some - I mentioned peudomonas earlier in this thread surveying in betadine - which is routinely used as a surgical scrub in the US.

 

[ScottB]

I did not agree with @jda that this discussion would bifurcate this way but sigh, he was correct.

Absent any information coming from the manufacturer, it is my opinion -- based on the evidence delivered -- if it waddles, swims, quacks, flies and looks like a duck then lets just treat it like a duck. (Or AlgeaFix in this example.)

I happen to like duck, but only when it is professionally prepared. I am equally sure AlgaeFix/Vibrant has a proper place in this hobby. We only should attach to it the proper risks that a stated/probable algaecide should be assigned.

My initial speculation in this thread was shredded early so I take no offense at tests (not conjecture) that prove a different thesis.

Occam's razor anyone?

 

[MnFish1]

I did not agree with @jda that this discussion would bifurcate this way but sigh, he was correct.

Absent any information coming from the manufacturer, it is my opinion -- based on the evidence delivered -- if it waddles, swims, quacks, flies and looks like a duck then lets just treat it like a duck. (Or AlgeaFix in this example.)

I happen to like duck, but only when it is professionally prepared. I am equally sure AlgaeFix/Vibrant has a proper place in this hobby. We only should attach to it the proper risks that a stated/probable algaecide should be assigned.

My initial speculation in this thread was shredded early so I take no offense at tests (not conjecture) that prove a different thesis.

Occam's razor anyone?

Well - I guess I could say - you were the one that thought it was fluconazole... But - In any case - in absence of comment from the manufacturer - There is no way to know - and the debate is tiring - and useless. The testing done is laudable. But - flawed. The company - which responded earlier - and not now - is flawed. So - I agree with you leave it this lol.

 

[taricha]

The testing done is laudable. But - flawed.

Look, if any test ever actually convinced you of anything, I think I'd disappear from from the forum due to heart attack.
It is unnecessary to update us like 5x on every page that you wish to discount all the testing done and you remain unconvinced. We know that already.

 

[taricha]

This next result is a bit of a surprise (depends on your expectations at this point I guess) and has implications for the question of why some tanks do poorly with Vibrant (It rules out a possible explanation).
@jda and others have suggested that Vibrant probably contains a strong dose of some organic carbon. The label aspartic acid and Vinegar amounts are not really enough of a carbon dose to do much of anything.

This is a Chemical Oxygen Demand (COD) test [hach has better documentation for the interested]. It uses strong oxidizing agents and heat to break down essentially any organic carbon. Any organic carbon that could have any effect on lowering nutrients in a system (carbs, alcohols, amino acids, proteins, etc) would all be easily broken down. So would any foods or whole bacteria etc.

AlgaeFix in Blue, Vibrant in Red, yellow is theory of the label carbon sources claimed in vibrant. (error bars are hanna stated uncertainty)

AlgaeFix and Vibrant have indistinguishable amounts of (chemically) digestable organics. For comparison, the yellow line is the theoretical result of what a solution of 0.5% Vinegar + 1% aspartic acid would give.
If Vibrant was an algaefix-like quat in a similar amount + any siginificant amount of organics, those lines would not agree so tightly.
Vibrant is not a significant organic carbon source. At this point it seems possible and even likely that it contains no carbon except something indistinguishable from the quat that is in algaefix, and the vinegar and aspartic acid is not actually present. This would be consistent with all the data produced so far.

(to be clear this chemical digestion can break down way more stuff than bacteria can, so just because it digests the quat in algaefix doesn't mean your bacteria can eat that as a food source. The algaefix polyquat is not a bacterial food source. A follow-up test can confirm this with letting bacteria "eat" each product and measure O2 consumption)

For those wondering why Vibrant causes bad results in some tanks, unfortunately - heavy carbon source is ruled out. It seems some tanks / organisms may simply have unpredictable sensitivities to the active ingredient. I got no better explanation for you at this point.