The Science Of Bottled Bacteria For "cycling" Aquaria
He makes a few good points in this interview:
  • The main one is the comment on the species of bacteria.
  • He also talks about heating and freezing. Here in Arizona it regularly gets 117 in the summer. A delivery truck that isn't refrigerated will easily exceed 140 inside the box. In such a delivery method the bacteria would be dead.
  • The third is that the bacteria needs to be fed but that doesn't address how they are fed in the bottle.
  • Finally, it's not an instant cycle. He states it takes 5-6 days which is more realistic than the claims some make of instant cycling.
  • Post Script: He challenges people to set up a tank and give it a shot. That's really the thing to the video (at the end) he does just that and tests it. The only question I have is that he puts 2 ocellaris in a 130 gallons (which I hope he pre-acclimated). Even without any additive those two fish would be fine in that size of tank because the PPM would be so small. least he experimented on it. I would be more interested in doing it on a 10 gallon tank.
I'm curious about this ABIL product by Avecom.
TTA has provided lots of links to scientific research which acknowledge ABIL as their starter inoculum to introduce nitrifying bacteria for various experiments.  In particular I like this one:
Page 1233:  "As nitrifying inoculum we used ABIL, in which the AAOB constituted on average 35.8% of the total DNA"
I find it surprising that they are using a product as a basis for their experiment seemingly without knowing exactly what's in it? 
Page 1229: "The presence of AAOB in ABIL and planctomycetes in the granular anaerobic sludge was confirmed by the real-time PCR count of both groups of bacteria"
...which gives me the impression they weren't entirely sure if ABIL contained any AOB to start with!
Presumably Avecom does not publish the contents of their product or the species of bacteria within it (unlike Dr Tim).  Yet it is still being used as a foundation for serious academic research, why?  I assume because it's already been proven to work in previous research papers, yet somebody must have been first to try it out!
Meanwhile Dr Tim has patented his breed of bacteria, which he has published research to prove they are the 'right' ones, and here we have ABIL which evidently also works yet neither of them are pressing lawsuits.  The bottles must contain different strains of bacteria which are equally (or equivalently) effective.  How then can Dr Tim claim that his are the right kind?
ABIL has left me rather confused so please shed light if you can! :)
Sorry daize- I posted the reply in the other thread. I had not seen your post here. So I am doing a copy paste-
daize- there is research re ABIL. Many research projects start off by using it to get a big jump on establishing the bacterial colonies they need for the research.

Improved performance of an intensive rotifer culture system by using a nitrifying inoculum (ABIL)

An improved nitrifying enrichment to remove ammonium and nitrite from freshwater aquaria systems
from http://www.sciencedi...044848601008833
Elevated salinity selects for a less diverse ammonia-oxidizing population in aquarium biofilters
from http://onlinelibrary...004.10.001/full
(this study identifies the bacteria)
2.4DNA sampling

Approximately 1 cm3 of polyester cotton from the middle part of the biofilters was removed and put in a sterile 10 ml screw cap vial. For the inoculum, 1 ml of nitrifying sludge was sampled. DNA samples were taken 1, 7, 14, 28, 35, 49 and 56 days after inoculation. DNA extraction  was performed immediately after sampling.
The fingerprint of the AOB community from the inoculum was dominated by one single band (Fig. 2, band a). The 16S rRNA gene fragment of 465 bp corresponding to this band has 99% (461/465 bp) similarity with Nitrosomonas sp. Is343 (AJ621032, data not published). In Fig. 2,
the DGGE fingerprints of the AOB community of all the freshwater and the artificial seawater biofilters at the start and at the end of the
experiment are shown. By sequencing it was confirmed that the dominant band from the inoculum remains present in all the aquaria for the entire time span of the experiment and even remains dominant in the DGGE fingerprint of the artificial seawater biofilters

Characterization of an Autotrophic Nitrogen-Removing Biofilm from a Highly Loaded Lab-Scale Rotating Biological Contactor
Start-up of autotrophic nitrogen removal reactors via sequential biocatalyst addition
from https://biblio.ugent...file/869933.pdf
Development of nitrifying bacterial consortia for immobilizing in nitrifying bioreactors designed for penaeid and non-penaeid larval rearing systems in the tropics
from http://nopr.niscair....(3) 240-248.pdf
Dimethyl sulfide removal from synthetic waste gas using a flat poly (dimethylsiloxane)-coated composite membrane bioreactor
from http://www.aseanbiot...ct/21015452.pdf

If you want to find more references do a Google Scholar search using "nitrifying bacteria inoculum + ABIL" and "nitrifying bacteria inoculum + Avecom"
It is a rare occasion where I post stuff that I have not already done a fair amount of reading of the available research so that I have a reasonable certainty that I am on solid ground.. There is no way I am going to mention a product like ABIL without having done so

The thing is, despite the research of Dr. Hovanec et. al., there appear to be a number of Nitrosomonas strains which are not identical but still seem to be able to do the same job.  if you look at the taxanomy here you will spot IS 134 and then look for Nm143- this is another one found to do a similar job. My impression is the bacteria are highly adaptive to specific locations and environments and that there are likely a lot more strains out there that have yet to be identified.
However, because it is possible to patent strains, for there to be more bacterial starter products out there they will need to utilize similar but not identical strains. This appears to the be the case with ABIL for example.

I understand, so it is possible for other products to effectively cycle a filter as long as they contain similar strains of bacteria.  If ABIL has a different 'right' kind of bacteria then there is potential for other existing or future products to work effectively as well.
Curiously, Bio-Spira has been around since before most of those articles were written so I wonder why they chose ABIL over a product whose contents were known.
The answer is simple really. Many of the studies were done in Europe which is where Avecom is located. In today's global world quality research is being conducted throughout the world.
Bio-Spira is now only a sw product. Remember, the AOB differ in sw and fw. That study which identifies the ABIL bacteria was done to determine just what bacteria worked at different salinity levels. And I am sure there are also patent and similar legal issues involved.
Yes- I am sure there will be more strains of Nitrosomonas discovered which will do a similar job to the ones in DrTim's product or Avecom's.
Consider the acid water situation. We know under normal circumstances the tank bacteria do not do well when the pH starts to drop under 7.0 and that somewhere they will stop working and even die. We also know if you pour the effective live bottled nitrifiers into water with an acid pH, the odds are good it will kill them. Yet we also know that "cycling" can and does occur at pH levels just under 4.0 and that similar bacteria are found to be doing the job.
There are a few things that make this pretty fascinating. For one, at those pH levels there is virtually no NH3 which is the preferred substrate for those AOB. The research indicates that when the bacteria adapt down to the really acid levels, they seem to be able to process either ammonium or perhaps urea. they are still the same strains but they seem to function differently. What all of this tells us is that the bacteria are pretty adaptive on both a macro and micro level.
And finally, lets not ignore the ability of species to adapt/change over generations. For people that means 20 years or so but for bacteria its much shorter. In fact we tend to forget when we discus this topic that bacteria is a collective term. The colonies are made of a numerous individual bacterium. Now while any individual bacterium may have a relatively short lifespan, the colony itself is durable over time. This means bacteria are able to adapt or evolve over many generations but only a very short amount of time.
Then lets consider the research that Dr. Hovanec et. al. have done. They took sample from tanks and went from there. They did not start out by saying this is what is there and lets go find it, they said something is there lets go find out what it is. But, realistically speaking, what they found was what predominated in the tanks they sampled at the outset. What I wonder is had it been possible to clone the researchers and to create several identical groups and then disperse them to different countries around the world, would they all have discovered identical strains or would there have been variation?
As we can see from DrTims and ABIL- two different but closely related strains are being used. What I draw from these sort of things is that even before one needs to identify the specific strain doing the job, one looks at more macro issues. For example, we know there are a variety of micro-organisms that can oxidize ammonia. Some are autotrophic, come heterotrophic, some are aerobic while others are anaerobic. So, even before one tries to identify the specific species and strains, there is the issue of determining which of the more macro qualities are relevant.
In tanks, it is obvious, to me at least, that the primary nitrifyers handling cycling are the autotrophic non-sporulating bacteria. They are what is in Bio-Spira, DrTims, SafeStart, ABIL etc. They are what are in the media we transfer for a fishless cycle. Now this doesn't mean there are not other bacteria in an established tank. Nor does it mean that products which claim to help establish the cycle do not contain bacteria that will initially control ammonia and nitrite thus giving the impression of establishing a cycle. But this omits the other piece of research need to prove that what is in the bottle is what predominates in the tank once it is established and running well.
There are only two studies I have found that show the bacteria at the start were still there down the road are Burrell et al in 2001 and the study on ABIL above identifying the species and strain. All of the other supporting data the makers provide is not lab grade research pulished in a peer reviewed journal. They are short term studies which only measure ammonia and nirtrite over the initial cycling period. They do not test for the presence of any other nitrifying bacteria during the few weeks involved and they do not test for anything beyond that. How does anybody have a clue if what is in Stability, API's or various other products is actually thriving in tanks and being the predominant ammonia and nitrite oxidizers?
What I can see is researchers using ABIL in the lab, large public aquariums using DrTim's in huge mission critical situations. I can not find this being done using Stability, Quick Start, and the other products for the Aquarium industry. While it may be easy to convince fish keeping hobbyists a product will help, it is another to get huge scale multimillion dollar businesses to do so easily. These rely on their own highly trained in-house experts or similarly skilled consultants for this.
Lets face it. If folks like us spend a few bucks on the product and use it with fish in and the worst happens- the ammonia and/or nitrite spikes and kills our fish, are we going to sue? It isn't worth it and would be absurdly expense to prove the product was at fault. On the other have if a major Public Aquarium or a huge multimillion dollar aquaculture relies on a product and the result is a massive fish kill, the outcome is going to be way different. And this difference is easy to see. There are a lot of small fish sites all over the net and fish keepers galore are posting on them that bacteria starts do and don't work. But I defy folks to find anything out there where an aquarium or fish farm has spent big money on bacteria, had if fail which resulted in massive losses.
When a huge public aquarium says they used the stuff and then added pricey livestock right away and that the ammonia and nitrite levels were minimal and gone very fast, and that no damage was done to the the tank inhabitants, are they lying? When multiple researchers state they used ABIL to jump start bacteria colonies needed for their study, is this a conspiracy or have they all been hoodwinked? Are they all misreading their instruments? Because if not, then there has to be something to the products being used over and over.
When I see the director of a major public aquarium say the product works, when I see credentialed researchers use it repeatedly and then I see some screen name on a fish site assure me it can not and does not work, guess who I am going to believe. I think it takes a complete lack of common sense to believe the reverse.
TwoTankAmin said:
When I see the director of a major public aquarium say the product works, when I see credentialed researchers use it repeatedly and then I see some screen name on a fish site assure me it can not and does not work, guess who I am going to believe. I think it takes a complete lack of common sense to believe the reverse.
The idea of good research is that we can ignore who is saying it and focus on what is being said, how it is being said, and how that conclusion was reached. Many great discoveries have been made by aquarists and many aquarists have gone on to be leaders in the field.  Think Martin Moe, Julian Sprung, Takashi Amano, Eric Borneman. The focus should be on the science not the personalities. "Some screen name on a fish site" can often have more practical experience and research time than someone with a degree in marine biology and there's no telling just who they are and what they know.
We all know that there are differing levels of experience on message boards like this one but I think it's a mistake to dismiss their input out of hand. Breeding techniques, new lifeforms like the water bear, have been discovered by mere hobbyists. Remember it was Norbert Tunze, working as a repairman who created the first powerhead. He was turned down for the idea by Eheim who saw no future in powerheads but did it on his own.
Anyway, my point is I don't like to see the hobbyist so easily dismissed.
I'm not interested so much in the fact that major public aquaria are using a product but why they are using it? why they chose it over other products? and what are the results? For example, they may have chosen it over another product due to cost or availability. Perhaps there is a better product but it costs more or because the ability of the company to manufacture it on a large scale was not in place so they could not meet the needs of a large facility. Perhaps there were import restrictions.
It's not enough to just say so-and-so uses brand-X so the other products must be crap. Perhaps there were other reasons.
I believe it's time to test this out for myself. I'll see if I can find an empty tank and give it a go. Keeping the above soapbox in mind as to reliability of the results. ;)
D.r Hovanec was a hobbyist who was so interested in the hobby that he got graduate degrees in related fields. Nobody on any forum where he posts nor at any function at which he speaks doesn't know his credentials. Anybody on this forum who has similar credentials and research experience is certainly permitted to say so. I know that Chris Cow another early proponent of fishless cycling has  Ph.D. in chemistry. I know that Stephan Tanner who sells the Poret foam among other things is a Ph.D, microbiologist. There are other hobbyists who have pretty impressive credentials and are not ashamed to say so on forums where they participate. Credentialed experts who actually post on forums do not hide the fact that they have their degrees and experience.
The thing you hit on the head is the part about the science standing alone. Go read the research papers and then decide if they stand alone. You will see the methods, the equipment, the results and the analysis of them spread out in plain site. And then you will see the conclusions of the research and a list of cited references. Where is all that from the forum members posting something works or does not, where are their methods, controls and multi-million dollar pieces of equipment? Laboratory research is conducted in a very controlled and precise way which is why mere hobbyists do not make these sort of scientific discoveries. For one to able to say a product does not work, they must be able to prove this is the case every time it is used. Further, nobody else should be able to have it work for them either.
Martin Moe Jr. is a credentialed scientist whose entire life was dedicated to the field of marine biology. It is his profession, not his hobby. If he comes here and posts on those topics, I sure as heck would listen, and if a user disagreed with him, do I have to tell you who I would  believe? Julian Sprung is in a similar vein, he has the undergraduate credentials and hands on experience. And he makes his living from his training and experience. It is his profession not his hobby. Eric Borneman also has the sw credential having been on his way to his Ph.D. But then he got arrested. I never saw how that ended up. He seems to have dropped out of sight? But until then he made decent money from his formal education and experience in sw. It was his profession not his hobby. Amano is a photographer, aquatic gardener and a designe. But if he comes here and wants to talk about microbiology and nitrification or fish biology, his credentials are mostly worthless in that respect. But he is also a professional in the aquarium trade, he is not simply a hobbyist.
While you are correct that hobbyists may have some extensive knowledge based on experience, to be a trained Ph.D. in chemistry or microbiology is not something one can do in their basement. Most of the people you called hobbyists above are actually trained professionals in fields directly related to various areas of the hobby. There is a great deal of difference between being a hobbyist and being a trained experienced professional.
If anybody on this site has a graduate degree in chemistry or is a microbiologist and they havein depth first hand experience with these things, they would be posting a lot differently than most of us can or do. None of the people you mention above did anything serious while they were only hobbyists, they did them after much schooling and years of hands on experience. Saying they are mere aquarists is basically a canard. So I will stick by what I said. Those people I choose not to believe on this (or any other) site are those saying things they do not back up with references. They simply state them as fact and offer no proof. I will only accept this from somebody with the credentials to make such statements legitimately. And credentials mean both degrees earned plus the hands on experience over time.
I'm not interested so much in the fact that major public aquaria are using a product but why they are using it?
One thing  should be obvious. The people who chose to use it must have believed it would work. These people are well trained and highly experienced in their field. All the other factors you try to bring in are not relevant. I do not think that you are suggesting that such extraneous factors would cause these people to spend one penny on anything they felt did not work. Nobody says the product that works costs too much, so instead we will save money and buy something much cheaper that does not work. We can't import the stuff that works, so we will buy the local product that doesn't. That would be beyond silly.
What we do know is what they actually did choose and that it did work. We know this because it is not hard to find out if that huge display tank went haywire and there were issues with cycling that harmed the dolphins or other tank inhabitants. The press would have been all over that as would a variety of animal activists etc. And then lets not forget that its not just one public aquarium that used the product, its is multiple facilities in more than one country. That doesn't make much sense if the stuff did not work. Sooner or later the word gets out among the industry insiders and nobody would be using it.
I can find references to people using Dr Hovanec's bacteria, I can find references to people using Avecom's ABIL, I can find references to folks using a few other things. But I can not find anything similar for Stability, for API Quick Start for example. I can research commercial starters for aquaculture and find even more bacterial products that folks in this hobby have never run across such as Novozymes PondProtect®.
And who claimed that just because a given product works it means that all others do not? Please reread what I have said and you will see that my beef with Stability, for example, is based on the fact that I believe the research which says that what handles ammonia and nitrite oxidation in tanks are autotrophic bacteria. Since I believe this, I can not then think that a product which states it contains none of these can be efficacious.
TwoTankAmin said:
One thing  should be obvious. The people who chose to use it must have believed it would work. These people are well trained and highly experienced in their field. All the other factors you try to bring in are not relevant. I do not think that you are suggesting that such extraneous factors would cause these people to spend one penny on anything they felt did not work.
We'll just have to disagree on the "knowledge of amateurs" for much great study, invention, and discovery have been done by them in all the disciplines. Though I am very much an academic I do not fall into the trap of "academic hubris" which brushes aside experience and wisdom as somehow less if it comes from one without letters.
As for the point of my latter statement, that the user believes the product works is a given in my comment above. I was speaking comparatively. If there are 5 products that work then we cannot assume that the one which was chosen was chosen because it was the "best". A product that is less effective could still be chosen based on a number of other business considerations. My point was to illustrate that while we may personally feel comforted by the fact that so-and-so uses a product this is not a scientific criteria that should be used when determining efficacy. Like my question #6 of page 1 I feel who uses a product is more akin to a celebrity endorsement rather than a genuine scientific aspect of the product. We may buy a set of golf clubs because Tiger Woods uses them but that doesn't necessarily translate the way we hope it will.
tcamos said:
I believe it's time to test this out for myself. I'll see if I can find an empty tank and give it a go. Keeping the above soapbox in mind as to reliability of the results.
I can add my own recent anecdotal experience with Dr Tim's One and Only.  I had a tank which I was unable to get a cycle started for about 10 weeks (I still don't know why, possibly because I used the wrong products or dosed the wrong amounts).  I then switched to Dr Tim's ammonium chloride and One and Only and the tank was cycled from start to finish in about two weeks.  That's with some difficulties with me reading results and following instructions along the way.  So from my personal experience I would definitely recommend Dr Tims as a good product.

It's the reason I now have fish in my signature :D
I am not trying to endorse Dr. Hovanec's products over others simply because he is the manufacturer. I am not trying to say that that experience has no value nor that merely having the letters next to one name in and of itself says anything about that persons specific skills vs those of a person without the letters but with a lifetime of hands on experience. But you are talking about a very small number of people. Moreover, the people I am quoting are not just people with a few letters after their name. Anybody who wants to can research them. I have not only looked into the education, public research, university affiliations etc. for Dr, Hovanec, but I have also done so for the other names on the three papers Dr H co-authored that I often reference.
So let me challenge you on this in a different fashion. There are private commercial labs and university labs and even some charitable organization labs all over the world that do genetic testing for research. They do the most sophisticated research into micro-organisms. So if your point is valid, you should be able to name at least one legitimate operation where just one of the senior researchers has no formal education and no degree in a relevant field. They are in that position solely based on experience. Or show me the gifted amateur who has spent several million dollars to purchase the same equipment so they can learn on their own in their basement or garage.
There is a big difference between somebody who spends years in the wild studying fish and habitats vs the trained biologist who takes some of those fish into a lab and studies the chemistry and biological processes of how ammonia effects the brains of those fish. There is a difference between somebody like me who has fishlessly cycled about 60-70 tanks over the years and a trained microbiologist who has done the genetic research to determine what specific bacteria were involved. I can tell you a lot about the mechanics of doing the cycle, I can suggest products to use and products to avoid based on my experience. But I still can not tell you what the very specific chemical interactions and genetics are. For that I turn to the experts.
I would ask you this. This site indicates it has close to 70,000 members. Lets say that only 25% of them are actually active for the sake of argument (it could be more or less). So that would mean around 17,500 actually stop in at some point in the year and read and maybe post. So of those 17,500 exactly how many have a graduate degree and/or work extensively in one of the scientific fields most relevant to our discussion? How many are professional chemists, biologists, zoologists etc. with or without the degree? How many of them are actually qualified based on the education and/or profession to have the experience to make the statements they do?
But lets consider your point. You gave me a bunch of names in your post. Almost all were experts in sw issues, a side of the hobby in which I do nothing and know very little. So I was not familiar with any of the names besides Amano. However, I was able to research each and every name. I could see what degrees they had, where they got them, I could see their work experience or the aquatic business they own as well. All it takes is a real name and maybe a tad more info, like a study or book they authored etc. I can not do this with a screen name. However, on more than one occasion on this site I have challenged folks on this very point. I have said I am basing what I say on this expert and that expert and then given links to their work. I have then asked the poster to offer to tell what degree or hands on experience they have that would make their statement to the contrary carry equal weight. It is simple for one to say I have a masters in biology or I have worked as a lab assistant for 25 years etc etc. I am still waiting for any such "closet scientist" to step up here. Nor would such a member have to post in an open thread if they are reticent, they could drop me a PM and tell me something like: "I am Dr. John Doe, I have degrees in biochemistry and have conducted both field and lab research into ....."
Last question for you tcamos. Who do you honestly think can most likely conduct a controlled experiment to determine what exactly is in say Satbility, Dr Tim's and API Quick Start in order to compare them? Who is able to say with the certainty that comes from doing the actual testing, what is in the bottles and if worked and then how effectively- lab trained, degreed scientists in a research setting or anybody you can select from the member list on this site?
As you said research isn't about the names, it is about the validity of the methods and results. At its heart what makes any results obtained in a controlled lab experiment valid is the fact that the methods are correct and that if somebody else replicates the work, they will get the same results every time.
I have to agree with TwoTankAmin.  Meaning no offence to experienced hobbyists who are undoubtedly very knowledgeable and have an accurate 'gut feeling' on many things, but I suspect on the subject of product efficacy hobbyists have nothing to offer except anecdotal evidence which cannot stand up to scientific scrutiny.   That includes me and my own anecdotal tale too! 
I'll also add that having a relevant degree is a very poor indication of expert knowledge and I would only really put faith in a professional or researcher in the field.  For example, I have a degree in engineering but I never pursued a career in that field.  My husband has a degree in chemistry, but he didn't even know what the nitrogen cycle was when I first asked him. :)
The thrust of my point is that any a focus on endorsement leaves off from science and into marketing.
The focus of this thread should be the studies done, how they were conducted, and who has repeated the work for verification rather than who has done it and who is using it.
The "cause" would better be served simply by labeling each product with a simple letter, Product A, Product B, and leaving off personality all together.
Tcamos- I posted a video on this site which was shot by the Georgia Aquarium. AT&T has their name on the opening credits. A part of this video shows the aquarium staff pouring very large buckets of Dr.Tim's One and Only into their immense dolphin tanks. They explain they are setting up the bio-filtration system which is crucial. So lets start with the assumption that the video is a not fake and that the events it depicts actually happened. Then let me ask you a couple of Qs relative to that video:
1. Why do you think the aquarium is putting a biological starter of any sort into the system?
2. Why do you think they are specifically using Dr Tim's. What does it say about his product?
3. Do you think the product worked for them as expected or do you think it failed to work for them with all the concomitant consequences?
daize- I would bet dollars to donuts that despite the fact you never became an engineer by profession that you know more about the subject than 99% of the membership on this site, including me. Also, the nitrogen cycle is more of a biological process than a chemical one at its heart. Give your husband a break.But he should be able to explain water chemistry to you and be a big help regarding test kits too.
I believe you are missing the point I am making. I am not sure I can make it more clearly than I did in my previous post.
I can see both sides I think.  Tcamos' point is that we should rely on the scientific research and not be concerned with who has used the product.  TTA's point is that large-scale commericial endorsement is a much better indicator of how well a product works than individual consumer anecdotes.
I'd like to agree with Tcamos.  It would be nice if there was valid independent research conducted that compared all the available products under controlled conditions to find out how well each one works.  If such research existed we wouldn't be having this discussion at all!  Instead the next best thing we've got is performance of the product under high-stakes commercial usage.  It's not ideal but it is certainly a more reliable indicator than the experience of individual hobbyists, so cannot be completely dismissed.
TwoTankAmin said:
Give your husband a break.But he should be able to explain water chemistry to you and be a big help regarding test kits too.
Haha the only help he's been is to express horrified concern that I'm not wearing goggles, gloves and lab coat while handling test chemicals! :lol:
But to illustrate my point, I told you he has a degree in chemistry and you've instantly credited him with expert knowledge in the field.  What if I now told you that his degree is actually Chemistry with Archaelogy and he only took the chemistry because it was the only way he could get on an archaeology course?  He never really had much interest in chemistry and can hardly remember any of it after 18 years, whereas I was very keen on chemistry at A-level grade.  I can still often put him to shame on chemistry general knowledge.

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