After manipulation and processing, these get very diluted down, which basically takes almost all the color with it as these serve such a small function in Vibrant as they do not have to have a strong presence. Totally opposite of a sludgy started bacterial product or any colored bacterial supplement in regards to this.Hi! I think I have polyglutamic acid, a similar anionic amino acid polymer. I can verify if this material can produce a color change. I highly doubt they will since both polyQ and polyD are both negatively charged, chemically similar and are unlikely to chelate the negatively charged dye. Heck, I'll even check some synthetic DNA today.
Nextly, I doubt there are cyanobacteria are in vibrant since the solution is clear and are not colored as cyanobacteria tend to be. In terms of bacillus, I guess they could be in there but the inactive spores of bacillus wouldn't be undergoing metabolism and probably wouldn't be producing these polyAAs, furthermore it wouldn't explain the algaecidal properties of vibrant since flocculants already exist for aquariums and they are not algaecide products. Although, to my understanding they could aid in the clearance of green water.
The pH of the BPB solution is controlled with excess phosphate buffer such that the pH contribution of any added analytes does not shift the color. Furthermore, the BPB colorshift to basic pH does not occur. @taricha has performed this test. Finally, the pH of both algaefix and vibrant are slightly acidic not basic.
In regards to your method, the method we both used was adapted from literature. But there are multiple ways of employing bromophenol blue to the detection of quaternary amines with or without chloroform extraction of the final complex. Basically as long as you add excess buffer and apply the solution to QAC, a color change occurs. No extraction is needed unless downstream analysis of the QAC is desired. OR if the analyte is in an unfriendly solution like milk. Considering that vibrant is clear as water. I don't think this is the case. Here are some literature (you can PM me if you'd like access):
Poly-γ-glutamate-based Materials for Multiple Infection Prophylaxis Possessing Versatile Coating Performance
Poly-γ-glutamate (PGA) possesses a nylon-like backbone and polyacrylate-like carboxyl groups, and shows an extraordinary solubility in water. In this study, the effective synthesis and structural analysis of some water-insoluble PGA ion-complexes (PGAICs) using cationic surfactants...www.mdpi.com
Colorimetric detection of residual quaternary ammonium compounds on dry surfaces and prediction of antimicrobial activity using bromophenol blue - PubMed
Controlling and monitoring the residual activity of quaternary ammonium compounds (QACs) are critical for maintaining safe yet effective levels of these agents in the environment. This study investigates the utility of bromophenol blue (BPB) as a safe, rapid and user-friendly indicator to detect...pubmed.ncbi.nlm.nih.gov
Towards an antimicrobial ‘microglove’ - Scientific Reports
A large proportion of hospital-related infections are acquired and spread due to the direct contacts between patients and healthcare workers. Accordingly, proper infection prevention measures and especially hand hygiene, are key to limit the spread of infections in nosocomial settings. However...www.nature.com
In the end. Vibrant likely contains quaternary ammonia or some other unlisted, highly charged compound capable of binding anionic dyes. Either way, its activity probably has nothing to do with anything listed on the label, in my opinion
My speculation: Vibrant has some fluconazole in it...
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MnFish1
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Hi! I think I have polyglutamic acid, a similar anionic amino acid polymer. I can verify if this material can produce a color change. I highly doubt they will since both polyQ and polyD are both negatively charged, chemically similar and are unlikely to chelate the negatively charged dye. Heck, I'll even check some synthetic DNA today.
Nextly, I doubt there are cyanobacteria are in vibrant since the solution is clear and are not colored as cyanobacteria tend to be. In terms of bacillus, I guess they could be in there but the inactive spores of bacillus wouldn't be undergoing metabolism and probably wouldn't be producing these polyAAs, furthermore it wouldn't explain the algaecidal properties of vibrant since flocculants already exist for aquariums and they are not algaecide products. Although, to my understanding they could aid in the clearance of green water.
The pH of the BPB solution is controlled with excess phosphate buffer such that the pH contribution of any added analytes does not shift the color. Furthermore, the BPB colorshift to basic pH does not occur. @taricha has performed this test. Finally, the pH of both algaefix and vibrant are slightly acidic not basic.
In regards to your method, the method we both used was adapted from literature. But there are multiple ways of employing bromophenol blue to the detection of quaternary amines with or without chloroform extraction of the final complex. Basically as long as you add excess buffer and apply the solution to QAC, a color change occurs. No extraction is needed unless downstream analysis of the QAC is desired. OR if the analyte is in an unfriendly solution like milk. Considering that vibrant is clear as water. I don't think this is the case. Here are some literature (you can PM me if you'd like access):
Poly-γ-glutamate-based Materials for Multiple Infection Prophylaxis Possessing Versatile Coating Performance
Poly-γ-glutamate (PGA) possesses a nylon-like backbone and polyacrylate-like carboxyl groups, and shows an extraordinary solubility in water. In this study, the effective synthesis and structural analysis of some water-insoluble PGA ion-complexes (PGAICs) using cationic surfactants...Colorimetric detection of residual quaternary ammonium compounds on dry surfaces and prediction of antimicrobial activity using bromophenol blue - PubMed
Controlling and monitoring the residual activity of quaternary ammonium compounds (QACs) are critical for maintaining safe yet effective levels of these agents in the environment. This study investigates the utility of bromophenol blue (BPB) as a safe, rapid and user-friendly indicator to detect...Towards an antimicrobial ‘microglove’ - Scientific Reports
A large proportion of hospital-related infections are acquired and spread due to the direct contacts between patients and healthcare workers. Accordingly, proper infection prevention measures and especially hand hygiene, are key to limit the spread of infections in nosocomial settings. However...
In the end. Vibrant likely contains quaternary ammonia or some other unlisted, highly charged compound capable of binding anionic dyes. Either way, its activity probably has nothing to do with anything listed on the label, in my opinion
You're absolutely correct that QAC's will change the color as you have shown. My point was that other things I believe can do it as well. So I wondered about pH, etc - I hadn't seen where you looked at that!!
In any case - You might be absolutely correct that its present. In my experience with the use of vibrant (which I have done twice) has been long-lasting. (granted - this was the freshwater version) - I had an extreme hair algae problem in a 200 gallon discus tank - which was wiped out - and has not recurred - for months- 1 year. Given the volatile nature of those compounds - it would surprise me if that were the active ingredient - especially since filtration, etc does not need to be changed and I believe carbon (and just detritus) would remove it quickly? Yet the effects seem to last much longer than the effective half-life.
Also - aren't those compounds quite toxic - i.e. I would be surprised that one could get enough of a concentration to kill algae without wiping out bacterial filters, etc - and potentially gill tissue. Having said that - I did have problems the one time I used vibrant in my reef tank - with various Acros so who knows.
The one sure thing with all of these 'compounds' - is one never knows exactly whats inside. But - I think there is a fair bit of evidence that 'bacteria' can outcompete algae. Whether those bacteria are in vibrant IDK. I can't remember - has anyone ever sent it into Aquabiomics?
Control Nuisance Algae Growth Naturally | Algae Control Canada
Our bioaugmentation process uses beneficial bacteria and enzymes to lower the high nutrient levels in water that cause nuisance algae growth and bad odours.
www.algaecontrol.ca
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Good points.You're absolutely correct that QAC's will change the color as you have shown. My point was that other things I believe can do it as well. So I wondered about pH, etc - I hadn't seen where you looked at that!!
In any case - You might be absolutely correct that its present. In my experience with the use of vibrant (which I have done twice) has been long-lasting. (granted - this was the freshwater version) - I had an extreme hair algae problem in a 200 gallon discus tank - which was wiped out - and has not recurred - for months- 1 year. Given the volatile nature of those compounds - it would surprise me if that were the active ingredient - especially since filtration, etc does not need to be changed and I believe carbon (and just detritus) would remove it quickly? Yet the effects seem to last much longer than the effective half-life.
Also - aren't those compounds quite toxic - i.e. I would be surprised that one could get enough of a concentration to kill algae without wiping out bacterial filters, etc - and potentially gill tissue. Having said that - I did have problems the one time I used vibrant in my reef tank - with various Acros so who knows.
The one sure thing with all of these 'compounds' - is one never knows exactly whats inside. But - I think there is a fair bit of evidence that 'bacteria' can outcompete algae. Whether those bacteria are in vibrant IDK. I can't remember - has anyone ever sent it into Aquabiomics?
Control Nuisance Algae Growth Naturally | Algae Control Canada
Our bioaugmentation process uses beneficial bacteria and enzymes to lower the high nutrient levels in water that cause nuisance algae growth and bad odours.
I ordered a bottle of Vibrant and will look at how it effects algae biofilm development versus no Vibrant. And for fun, I will look for a Microbacter Clean effect. This will add nothing to the ingredient discussion but might shed some light on the question of whether it is a biocide. I will eventually get around to looking at what it does to Bio-Spira biofilms.
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AlgaeFix marine explicitly contains one of these quaternary ammonia compounds as the ingredient, and in my tank it works great at killing algae - shows no noticeable (to me) effect on fish or inverts, and an aquabiomics sample done a month or so after I did several rounds of algaefix reported that my microbiome was "textbook normal".
So, yes. It's totally possible for these compounds to be used as an algacide in a reef tank and kill algae, and not nuke everything.
Regarding pH....
Approximate pH of AlgaeFix marine was ~6.1 and Vibrant was ~6.1-6.2
The dried residues of each was also slightly acidic, not strongly basic. Here's the dried residues with Phenol Red (yellow at pH 6.8 to Red at 8.4, comes out of the bottle red)
L to R: Vibrant, tap water, Algaefix
So the shift to blue color in this test isn't from high pH in the dried drops.
MnFish1
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Excellent. Thansk for that. By any chance - did you check the bromphenol blue test - with merely a solution with pH 11 or 12? Though the test you did was 'phosphate buffered' - buffers can be overcome. And - if there is an excess of buffer (which I believe was the case) - it can affect all of the results. But - in any case - thats probably splitting hairs - would you mind using the simple test-tube protocol that I gave (I think you have all of the ingredients) - and show the results - im just curious. The paper (which was actually very interesting) - is designed to show residual amounts of QAC - after killing bacteria. I'm not sure its the same as adding the chemical to an aquarium - and my main question is 'what other chemicals can cause a similar reaction'. If there is a QAC in vibrant (in high enough concentration to cause this reaction you showed) - I believe you're entirely correct - I do not think bacteria (or spores) can survive. But - as said before - there is also the question of vibrant continuing to work weeks after dosing - as compared to immediate killing (which is what I would expect) - in any case - I think we can all agree its not fluconazoleAlgaeFix marine explicitly contains one of these quaternary ammonia compounds as the ingredient, and in my tank it works great at killing algae - shows no noticeable (to me) effect on fish or inverts, and an aquabiomics sample done a month or so after I did several rounds of algaefix reported that my microbiome was "textbook normal".
So, yes. It's totally possible for these compounds to be used as an algacide in a reef tank and kill algae, and not nuke everything.
Regarding pH....
Approximate pH of AlgaeFix marine was ~6.1 and Vibrant was ~6.1-6.2
The dried residues of each was also slightly acidic, not strongly basic. Here's the dried residues with Phenol Red (yellow at pH 6.8 to Red at 8.4, comes out of the bottle red)
L to R: Vibrant, tap water, Algaefix
So the shift to blue color in this test isn't from high pH in the dried drops.
- which was the original topic
You are smart and I am not saying that you have not already concluded this, but set aside the compound that kills the algae and take a look at what else might be in the bottle. Known by many posters to quickly drop nitrate and phosphate, fuel dinos and some cyano, clear up some water, provide some fuel for some bacterial growth and cause some extra skimmer action. Some who have overdosed have reported oxygen depletion and heavy breathing fish in a few anecdotes. What other compound does these things that is known to tick off especially sensitive acropora when used too quickly or too much? ...especially one that if you knew was in there, you might not have used in your tank without more caution.
Knowing the looks and smell of what is in the bottle, you can narrow this down even a bit more. With AlgaeFix, once absorbed even immediately, it can take a while for stuff to die. I used some Quat in my septic pond for some duckweed and it worked for like 2 years and the compound is supposed to break down faster than that (according to label - like gone in a month?), but how much of that is that there was no duckweed left to reproduce anymore? I don't have any answers and I am not smart enough... just supposing.
MnFish1
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OH I think all of youre points make complete sense. And - based on my experience - I would not use it again in a reef tank - And - as I typed the last post I wondered if I just killed 'all the hair algae' - but that seems just appear by some miracle - whether its spores in the air etc. IDK. - But yes - I totally agree. (I also broke the command rule of reefing - nothing happens fast - I overdosed by using the 'higher dose' first - as compared to waiting a couple weeks. I do think its reasonable to think that bacteria compete (outcompete) algae - based on their faster reproduction rates - BUT - whether that can be bottled and reliably sold - that I'm not as sure. And PS - I'm not smart.You are smart and I am not saying that you have not already concluded this, but set aside the compound that kills the algae and take a look at what else might be in the bottle. Known by many posters to quickly drop nitrate and phosphate, fuel dinos and some cyano, clear up some water, provide some fuel for some bacterial growth and cause some extra skimmer action. Some who have overdosed have reported oxygen depletion and heavy breathing fish in a few anecdotes. What other compound does these things that is known to tick off especially sensitive acropora when used too quickly or too much? ...especially one that if you knew was in there, you might not have used in your tank without more caution.
With AlgaeFix, once absorbed even immediately, it can take a while for stuff to die. I used some Quat in my septic pond for some duckweed and it worked for like 2 years and the compound is supposed to break down faster than that (according to label - like gone in a month?), but how much of that is that there was no duckweed left to reproduce anymore? I don't have any answers and I am not smart enough... just supposing.
MnFish1
10K Club member
OH - BTW - all of the 'symptoms' you mentioned - suggesting oxygen depletion suggest 'bacteria' as compared to chemical. Not that chemical is impossible - but if it was 'chemical' - it would seem to me that as algae is rapidly killed (picture roundup on a lawn killing weeds), nitrate and Perhaps PO4 would significantly rise. Right?You are smart and I am not saying that you have not already concluded this, but set aside the compound that kills the algae and take a look at what else might be in the bottle. Known by many posters to quickly drop nitrate and phosphate, fuel dinos and some cyano, clear up some water, provide some fuel for some bacterial growth and cause some extra skimmer action. Some who have overdosed have reported oxygen depletion and heavy breathing fish in a few anecdotes. What other compound does these things that is known to tick off especially sensitive acropora when used too quickly or too much? ...especially one that if you knew was in there, you might not have used in your tank without more caution.
With AlgaeFix, once absorbed even immediately, it can take a while for stuff to die. I used some Quat in my septic pond for some duckweed and it worked for like 2 years and the compound is supposed to break down faster than that (according to label - like gone in a month?), but how much of that is that there was no duckweed left to reproduce anymore? I don't have any answers and I am not smart enough... just supposing.
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It's worth asking how much of this stuff is being detected in Vibrant, as compared to AlgaeFix where it is the main ingredient.
So I did dilutions of AlgaeFix and Vibrant each in my tank water (then did dried residues of 1 drop of each dilution) to see how low it had to go for each before the Bromphenol purple to blue effect stopped.
This is not a precise titration (which could be done), but part of the paper I'm working off of did use something similar to estimate a concentration.
The detection for both AlgaeFix and Vibrant occured at dilutions of 1/3, 1/10, 1/30, and 1/100....
So I went lower.
The effect was also detected at 1/100 (again), and 1/300, but not at 1/1000, 1/3000, or straight tank water. It appears the amount of quaternary ammonium compounds in vibrant and algaefix may be similar to within a factor of 3.
(Obviously diluting with tank water like this, pH is essentially the same across all of these except the pure AlgaeFix & vibrant.)
So I did dilutions of AlgaeFix and Vibrant each in my tank water (then did dried residues of 1 drop of each dilution) to see how low it had to go for each before the Bromphenol purple to blue effect stopped.
This is not a precise titration (which could be done), but part of the paper I'm working off of did use something similar to estimate a concentration.
The detection for both AlgaeFix and Vibrant occured at dilutions of 1/3, 1/10, 1/30, and 1/100....
So I went lower.
The effect was also detected at 1/100 (again), and 1/300, but not at 1/1000, 1/3000, or straight tank water. It appears the amount of quaternary ammonium compounds in vibrant and algaefix may be similar to within a factor of 3.
(Obviously diluting with tank water like this, pH is essentially the same across all of these except the pure AlgaeFix & vibrant.)
Unless there is both, right?
You can get those same bacteria based "effects" with sugar water, vodka, vinegar, ethanol and a myriad of other things that do not kill algae. My point was not that the bacterial based stuff did not make your acropora mad, but it was unlikely that the algae killing part did it. Does there have to be actual bacteria in a mixture for it to fuel some?
MnFish1
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Awesome experiment !!! - but imho its not obvious that diluting with tank water would mean the pH is essentially the same. Can you do me a big favorIt's worth asking how much of this stuff is being detected in Vibrant, as compared to AlgaeFix where it is the main ingredient.
So I did dilutions of AlgaeFix and Vibrant each in my tank water (then did dried residues of 1 drop of each dilution) to see how low it had to go for each before the Bromphenol purple to blue effect stopped.
This is not a precise titration (which could be done), but part of the paper I'm working off of did use something similar to estimate a concentration.
The detection for both AlgaeFix and Vibrant occured at dilutions of 1/3, 1/10, 1/30, and 1/100....
So I went lower.
The effect was also detected at 1/100 (again), and 1/300, but not at 1/1000, 1/3000, or straight tank water. It appears the amount of quaternary ammonium compounds in vibrant and algaefix may be similar to within a factor of 3.
(Obviously diluting with tank water like this, pH is essentially the same across all of these except the pure AlgaeFix & vibrant.)
. Use that protocol using bromphenol blue - and show the results? I am not totally sure that the 'dried' version is the same as detecting liquid versions. If you don't want to do it its fine - I was just interested in the result - and I dont have access to the chemicals
MnFish1
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Yes!! I guess I would HOPE - that when a company states '
95% Cultured Bacteria Blend
1% Amino Acids (Aspartic Acid)
0.5% Vinegar
3.5% RO/DI Water"
that in fact those are the ingredients. It seems to me if that NOT the case - that would be wrong. IMHO - I do not think that QAC are the active ingredient - but rather based on your own comments - some bacteria - that (as is common knowledge) - quickly outgrows algae - that is a spore-former - and rapidly grows in aquarium (especially dirty aquariums) conditions. Which to me is why vibrant takes 'days/weeks' to work as compared to hours/days. But - anything is possible. I do not automatically assume that published ingredients are a 'lie' - but - as I said before - anyone can say anything. What I dont understand (frankly) is the percentages used - 95 percent cultured bacteria blend - this does not make sense to me. If 95 percent was bacteria - there would be a lot more than 3.5 percent RODI water etc - (or it would be a dry cube) - so I don't completely understand the ingredients myself. No clue as to how they got there - and I'm not assuming they are made up - just dont understand
I am happy to be wrong given that my speculation kicked off this enlightened and friendly discussion.But - as said before - there is also the question of vibrant continuing to work weeks after dosing - as compared to immediate killing (which is what I would expect) - in any case - I think we can all agree its not fluconazole
I keep weekly nutrient measures and Vibrant certainly does lower them. The mix of bacteria vs carbon source versus anything else I cannot speak to. I was speculating off of what my experienced eyes told me regarding fluc and the need for photosynthetic activity. The shaded stuff stuck around for an month or more.
FWIW, I was actually dosing Vibrant to see if it would control a chrysophyte outbreak. It did that. But GHA then replaced that. I kept up only the maintenance dose afterward and it largely controlled that was well. Except for the shaded area. I've since replaced a few missing tangs/rabbits and all is clear now.
jeffww
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Here you go: ph9-6 and 0.1 M naoh(ph>12) oh and huge excesses of glutamic acid and aspartic acid. Also the panel I did the other day….seems pretty telling! Vibrant probably has quaternary ammonium! Also, polyquat is used to flocc water. Which explains the clarifying.
yesterday:
Ill also add on that if its something that “outcompetes” the algae but somehow also simultaneously doesnt grow in sea water seems contradictory. Lets be real here the makers of vibrant are not being totally truthful in their labeling one way or another. I would not recommend anyone put something in their tank like that.
yesterday:
Ill also add on that if its something that “outcompetes” the algae but somehow also simultaneously doesnt grow in sea water seems contradictory. Lets be real here the makers of vibrant are not being totally truthful in their labeling one way or another. I would not recommend anyone put something in their tank like that.
After seeing Boyd on RC a decade or more ago trying to mislead as to what was in Chemiclean and even going so far as to explicitly say that a similar compound Eurthromycin Succinate was not in the product to distract or mislead (or something like that), I don't trust anybody anymore. Nobody has to tell us anything, but I just wish that they would not lie about it. This kind of stuff can make people mad and nobody gets mad at the ones who don't say what is in their stuff. Is anybody mad at Kz because they don't tell us what is in Coral Booster?
Just a general, generic question to the masses... in a mixture with this amount of quat and some source of organic carbon in quantity to have an actual effect on a tank, could a live bacteria blend live in such a mixture?
MnFish1
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Excellent work - my eyes are not good enough to see the exact results (and lets face it your writing is close to mine - ie. hard to read lolHere you go: ph9-6 and 0.1 M naoh(ph>12) oh and huge excesses of glutamic acid and aspartic acid. Also the panel I did the other day….seems pretty telling! Vibrant probably has quaternary ammonium! Also, polyquat is used to flocc water. Which explains the clarifying.
yesterday:
Ill also add on that if its something that “outcompetes” the algae but somehow also simultaneously doesnt grow in sea water seems contradictory. Lets be real here the makers of vibrant are not being totally truthful in their labeling one way or another. I would not recommend anyone put something in their tank like that.
- But the colors I cannot really see. BUT - I would suggest - one problem is - that it could still be bacteria in the bottle - that reproduces once in the tank - the (excellent) chemistry aside - the chemistry results could be a result of another inert additive. I'm not saying it is - just that its possible. I do not see (from a common sense perspective) - how a QAC work given what I have read to be an extremely rapid decomposition in a dirty tank. Interesting results though
Spare time
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It doesn't have fluconazole. There isn't a reason to suspect it. The manufacturer thinks one of the bacterial strains may eat algae. Other than that its just a bunch of bacteria meant to outcompete algae and scavenge.
MnFish1
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Right - I would say 'no' - but I haven't researched it entirely. Spores of bacteria can be quite resistant. But - the issue here is we don't know the concentration of QAC - its a dehydrated version. (At least thats my reading) - that it may very well contain QAC - but its impossible to know 'how much'. What I still cant get my head around is how would QAC work - it is a general disinfectant - it is also quickly neutralized by organics. So - IDK - The chemistry is interesting. I might present the data to Vibrant and ask them? But - as I said before I do not understand the 'bacteria blend' of 95 percent. that do me does not make common sense either.
The quat in AlgaeFix has nearly the same instructions, results and timeline of killing and reducing algae. It is effective at that. Quat works in marine tanks, as @taricha indicated above. You can probably use this as a baseline fact.
jeffww
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The qac in algae fix is polyqac which is more specific to algae rather than bacteria. I don’t think I could do much more if the final argument is “I can’t see it” regarding the images. Ask yourself as a logical person. Whats more likely: some guy in a garage purifies some exotic strain of algae eating bacteria? Or just spikes it with polyqac, a common algaecide with proven efficacy and safety for aquariums.
