An Open Challenge To Ianho -

[OldMan47]

I have not read through the references you gave but am the first to agree that Tim Havonec at one time had the definitive research into the identity of the predominant AOB and NOB species. What in the world does that have to do with anything you are supporting today?
Around the time he first published his reviewed results, he entered into a manufacturing agreement that led to production of products called Nitrospira. It was a short shelf life refrigerated product that failed to perform well in about half the cases that we documented with actual experiences of people, after they had obtained the product commercially, not in lab conditions, but in the rest of the cases it seemed to provide some improvement over a simple fishless cycle the way we generally conduct one. I immediately concede that is true since we have plenty of evidence right here on TFF to support its partial success. Unfortunately, the failure in close to 50% of the cases led Marineland to stop producing the freshwater product because it was not a commercial success. People simply would not pay the high costs that were being charged to improve their cycle time marginally in large enough numbers to support its continued production. I hope this discussion is not about Nitrospira because I will end it right now by conceding that such a product did indeed once exist and did indeed provide a 50% probability of improving cycle times if it was cared for properly throughout its process from manufacture to final use. In fact I just bottled and sent off, today, a sample from my own filter that someone will try to use to jump start their cycle. I enclosed it in a double bag with a very small amount of water and lots of oxygen and sent it priority mail to minimize the time it needs to survive before being used. I fully expect it to survive the time of less than a week to reach its destination and am also ready to conceded immediately that a bacterial sample can survive in adequate numbers to be useful for a week if it is given enough oxygen to continue alive, even at room temperatures. Again, if that is your contention, I concede that you are right.
On the other hand you seem to be supporting nostrums like Cycle and Dr Tim's One and Only which are not refrigerated, do not get packaged with large amounts of oxygen and minimal amounts of water and have reputed long shelf lives. I await any evidence that you have that supports such a contention. The short quote you provided that we should be aware of from Cycle gives an example of adding a "bacterial starter" for 4 consecutive weeks. Any organic material added to a tanks filter for 4 consecutive doses at one week intervals will indeed promote some cycling activity, it is equivalent to using fish food to cycle a tank. I find it a bit less accurate way to cycle than using straight ammonia doses since the decay rates are less predictable but it will supply the needed organic nutrients. We get repeated reports from people who try use just Cycle that they experience a slow build in ammonia with no fish present, so I assume it contains some sort of organic material that decays and may even be a substitute for the ammonia that we would normally recommend.

 

[shrimply]

OK so I haven't really read the papers, I don't really have the time or motivation right now. Once I finish learning stuff I actually need to I might have a proper look through them. But few points coming from no ones side.

Firstly Ianho, One of your main arguments seems to be the funding/motivation for research, now leaving aside the file drawer problem which can only really be identified by a meta-analysis. It really doesn't matter who carried out the research or why, if the papers are published in a scientific journal they will be thoroughly peer reviewed. There for short of them actually making up the results its reasonable to assume the conclusions they come to are correct, or at least conclusions that may be wrong are worded in a way that they are personal speculation not fact.

Secondly I agree with previous comments the papers seem to be about identifying the bacteria, while the argument resolves around the successful long term storage of the bacteria.

And finally, for now, I need to go learn about deep homology (Yay), these papers are pretty old, I haven't read through the techniques much, but I imagine giving the advances of PCR and genotyping and the tree of life project, You could carry out a really nice and much more detailed study to find out exactly what is in filters. Taking samples from filters around the country and simply sequencing everything in them, then comparing to the online databases. In much the same way as environmental sampling works.

This still wouldn't tell you whether or not they survive in a bottle though... And although maybe not a great comparison when you consider the hassle that comes when trying to use a live vaccine and keeping the product alive long enough for it to be affective you do have to wonder how long bacteria, which as far as I'm aware does not have a dormant state can survive in a bottle with limited food etc.

 

[ian]

Firstly Ianho, One of your main arguments seems to be the funding/motivation for research, now leaving aside the file drawer problem which can only really be identified by a meta-analysis. It really doesn't matter who carried out the research or why, if the papers are published in a scientific journal they will be thoroughly peer reviewed. There for short of them actually making up the results its reasonable to assume the conclusions they come to are correct, or at least conclusions that may be wrong are worded in a way that they are personal speculation not fact.

good point Shrimly, but i have to disagree regarding the 'who did the research' comment. It makes all the difference to who actually does and funds research. Theres quite a famous article about cranberry juice and the releif in cyctitis. This article was taken on bored by medics and the likes (even myself working in a GP practice, not as a gp i add)...the research was later to be found that it was funded by a subsidery of ocean spray, the biggest harvester of cranberries in the world. Hence the sale of cranberry juice went through the roof. When it was found that the subsidery was actually owned by the Ocean spray, subsiquent research was done 'independently' and it was found that in fact Cranberry juice was no better than plain old orange juice. This is why (especially for me) you should approach ALL research with a very open mind, and look around the research first before diving staright in and believing everything thats written.

This is also the way you 'should' be tought to approach research at university level. I have tought student nurses in the past and always advised this approach as does univerity scholers and professors.

This is why when i see Dr Tims name against any research, you have to think of the gains he's going to get or getting from the research proposed. I'm also unsure whether the research is peer reviewed?? I can't see that it is?? It doesn't have to be peer reviewed to go into a journal (it always helps though)

as said i don't 'not' believe the research done in the article i have read, it just doesn't back up any previous discussion had between myself, oldman and TTA.

right oh back to work...

 

[shrimply]

Indeed, at least in biology a large part of the teaching is how to analyse papers, and to actually think about what is being talked about not just take what is written as fact.

Regarding Peer Review, my understanding (which maybe well be wrong) was that all journals at least in the scientific community would not publish papers without peer review.

Nature's website says
"Peer review is commonly accepted as an essential part of scientific publication"

That being the case, all methods and analysis/interpretation should be scrutinised by other independent scientists before it makes publication. But I don't know exactly what the situation is here or the peer review policies ( if any) of the journals in questions.

 

[Noahs ark6]

Who else is hoping Ianho leaves a conclusive comment then locks it that would be using mod privileges to there full potential

Bae

It would also be shying away from the confrontation, and i'm quite enjoying this, please don't lock it

 

[TwoTankAmin]

I think it would help this thread stay on topic a lot better if folks who have absolutely nothing to contribute would refrain from cluttering up the thread with irrelevant comments.


Atlas, I think you have missed the impetus of the research as well as the results. The use of Cycle was a secondary part of the research. They ran a bunch of tanks with the intent of trying to identify the bacteria doing nitrite oxidation in aquariums. They added Cycle, which they knew contained no Nitrospira but did contain Nitrobacter to one set of tanks. The testing showed there was some difference in how fast nitrite spiked in all the tanks not dosed with Cycle vs the 3 with it. But they also showed that despite a difference in how fast nitrite levels peaked, nitrite hit 0 at about the same time for both and both had similar nitrate levels.

By day 22, the nitrite value had reached a maximum in the tank which received the additive. Nitrite concentrations reached maxima in the nonadditive aquarium on about day 32. By day 38, the nitrite levels in both aquaria were essentially below our limits of detection, and nitrate levels were equivalent in the treated and nontreated aquaria (Fig. 8).

They merely speculated that since it was not the Nitrobacter which were doing the nitrite oxidation, that there must have been another reason and that maybe it was something else in the Cycle bottle. Lets bear in mind that in 1998 there was no Bio-Spira, no Safe Start and no One and Only. Any bottled bacteria back then would only have the actual strains by accident not by design.

Now, you and some other folks asked to see any other evidence that bacteria can survive in a bottle. Here is a 21 page review on this topic. It is the work of a consortium of 7 European companies in businesses related to aquaculture etc. This paper Section 2 - Methods for preserving bacterial cultures. Section 3- Storage of nitrifying bacteria. Section 4- Factors affecting the survival of nitrifying bacteria during storage (4.1 Ammonia-oxidizing bacteria - 4.1 Nitrite-oxidizing bacteria). This paper includes references to some 87 other studies, one of which is Hovanec et al. This can be opened in Microsoft word. It clearly shows that there are a variety of working methods for storing nitrifyiers as well as other bacteria.
http/adapond.eii.e...eliverable.docx

These are the organizations/companies in the consortium
Remedium (Coordinator) Estonia
Consul AR Switzerland
Aquarius Marine Group UK
Biopharma Technology UK
Aquamyk Estonia
CGS Italy
Biomar Denmark
Aqua Consult Ingenieur Germany
Eesti Innovatsiooni Instituut Estonia
Teknologisk Institutt Norway
AquaOptima Norway
From http/adapond.eii.e...out-the-project


Ianho- firstly

as, as above i have read the thread and i'm still unsure you're able to critique research properly as Atlas appears to have found loads of flaws already...

No, Atlas did not find flaws, as I explained above. But nice try, Ianho. Moreover if you do not know what a bioreactor is, I think maybe I am not the one not able to critique this sort of research. But let me help you out.

A bioreactor is a vessel in which a chemical process is carried out which involves active substances derived from organisms. This process can either be aerobic or anaerobic.
A bioreactor may also refer to a device or system meant to grow cells or tissues in the context of cell culture.
There are many research applications of a bioreactor and the environment inside may be controlled to attempt gain reaction or specific reaction depending on the research.
In simplest terms it is a complex terrarium to cultivate organisms for research.

From http/wiki.answers....n_of_bioreactor

And to give you an idea how common a bioreactor is in research, if you Google Scholar the word "bioreactor" (excluding patents), you will get back about 228,000 studies.

Plus, if you read the paper you will see this out the outset:

Operation of bioreactors.Laboratory-scale bioreactors were used to produce the enriched nitrifying biomass. The bioreactors were circular columns of clear polyvinyl chloride pipe 198 mm in diameter and 600 mm tall for a maximum volume of 18.5 liters. Each reactor had a lid to minimize aerial contamination and aerosol production. A magnetic stirrer and air diffuser served to maximize mixing and maintain the dissolved oxygen concentration above 5 mg/liter. The bioreactors were kept in darkened cabinets at 26°C. The influent comprised a simple autotrophic medium, free of organic carbon, consisting of potassium phosphate (0.5 mg/liter) and ammonium chloride. The ammonia N concentration in the enrichment medium was kept in the range of 5 to 10 mg/liter for the low-concentration ammonia reactors and 40 to 60 mg/liter for the high-concen-tration ammonia reactors (Table 1). Bioreactors were monitored daily and maintained at their predefined ammonia N concentrations by feeding the autotrophic media when required. Ammonia, nitrite, and nitrate concentrations were rou-tinely monitored byflow injection analysis and ion chromatography (12). The pH of the bacterial suspensions was kept at or above a pH of 8.0 through the addition of sodium bicarbonate. Fifty percent water changes were performed weekly by allowing the nitrifying biomass to settle, decanting and discarding the appropriate volume of supernatant, and replacing it with the equivalent volume of deionized water.

Now lets depart from the science for a second to address your comments regarding potential research bias because a product results. I am aware of this phenomenon. We know Bio-Spira was the first product to contain the bacteria from the Hovanec et al. research and that it was released in June 2002. But Dr. Hovanec et al. published their first work in August 1996 entitled "Comparative Analysis of Nitrifying Bacteria Associated with Freshwater and Marine Aquaria. Applied and Environmental Microbiology Vol. 62, No. 8: 2888-2896. Hovanec, T. A. and E. F. DeLong. 1996." (Note this paper was published before Dr. Hovanec had earned his PhD. but was employed by Aquaria, Inc. and the co-author E.F. Delong was a Ph.D Professor at UCSB). A full 12+ prior to Bio-Spira.

Just so you get a feel for the whole process which ends up with Dr Hovanec and One and Only, here is a timeline from Marineland showing chronology http/web.archive.o...ra_timeline.asp

More interesting is the purpose of the 1996 study was to take samples from fish tanks and find the "wrong" bacteria that everyone believed were at work in tanks- Nitrosomonas europea and Nitrobacter wyinogradskyi. They took samples from 38 fw and 14 sw tanks. A first for this type of research as it was directly related to aqquaria. What Hovanec and DeLong discovered was that, in fw tanks, the bacteria assumed to be the nitrifyers were not there. And the ones assumed to oxidize nitrite were nowhere to be found in any tanks. And this was the beginings of identifying the exact strains as well as showing the ones believed to be there, actually were not.

Ianho are you saying that in the early 1990s Dr, Hovanec was doing biased research so he could be selling One and Only starting in 2007 or so? Or are you saying he was began doing this in the 1996 paper? Or perhaps you believe this plot began with the 1998 study. And if not it has to be by the time the work for the 2001 study was being done as Bio-Spira came to market the following year. But of course he did not own/sell the product, Marineland did. Dr. Hovanec was an employee.

I just want to know, according to you, what parts of the research with Dr. Hovanec's name on it should be deemed suspicious and not valid due to the fact that the very end result after 15 years of work is his current product. I would also like to know how you are so savvy about this while dozens and dozens of subsequent researchers who cite these works were completely fooled. I want to hear from you why most of the names as co-authors who have remained in academeia or non commercial research are also guilty of biasing the results. These are mostly Ph.D.s, who, to the best of my knowledge have neither repudiates their work with Dr. Hovance nor have they published subsequent research which supplants their earlier studies. And almost none of them has any commercial interest. Only Phalen is a co-patent holder for one of the bacteria and methods for detecting it.

Finally, can you please explain how Dr. Hovanec managed to subvert other independent researchers who have confirmed his work such as Blackall, 2000; Okabe et al., 1999, Schramm et al. 1998. Or "Nitrosomonas Nm143-like ammonia oxidizers and Nitrospira marina-like nitrite oxidizers dominate the nitrifier community in a marine aquaculture biofilm" a 2007 study by a world class team of researchers' You can read the full study here http/onlinelibrary.wiley.com/doi/10.1111/j.1574-6941.2007.00418.x/full

1. Bärbel U. Foesel[sup]1,2[/sup],
2. Armin Gieseke[sup]3[/sup],
3. Carsten Schwermer[sup]3[/sup],
4. Peter Stief[sup]3[/sup],
5. Liat Koch[sup]4[/sup],
6. Eddie Cytryn[sup]4,5[/sup],
7. José R. De La Torré[sup]6[/sup],
8. Jaap Van Rijn[sup]5[/sup],
9. Dror Minz[sup]4[/sup],
10. Harold L. Drake[sup]2[/sup],
11. Andreas Schramm[sup]1,2[/sup]

[sup]1[/sup]Department of Biological Sciences, Microbiology, University of Aarhus, Aarhus, Denmark
[sup]2[/sup]Department of Ecological Microbiology, University of Bayreuth, Bayreuth, Germany
[sup]3[/sup]Microsensor Group, Max Planck Institute for Marine Microbiology, Bremen, Germany
[sup]4[/sup]The Volcani Center, Institute for Soil, Water and Environmental Sciences, ARO, Bet-Dagan, Israel
[sup]5[/sup]Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, Rehovot, Israel
[sup]6[/sup]Department of Civil and Environmental Engineering, University of Washington, Seattle, WA, USA


Ianho, this has to be some conspiracy spanning multiple decades, assorted universities and research organizations and literally many 100s of scientists. All designed to make Dr. Hovanec a few dollars from selling his bacteria.


Here is how I see it. On one side are decades of research by countless institutions and scientists all who have contributed along the way to get to the end result, viable bacteria in a bottle. On the other side I find you and some anecdotal results which involve mostly newbie fishkeepers who have more than likely failed to follow simple directions and thus failed to achieve the results the makers claim you should see for their products. I also find that similar anecdotal research which says the product worked for them is totally discounted by you. Double standards are certainly not scientific and are not even anecdotally supportable.

Oh yes, you also have some forums trolls who are supporting you with snide comments in this thread but who are contributing absolutely nothing scientific nor relevant to the discussion. It seems to me a moderator on this site should be working to keep such stuff out of scientific threads. I have read the guidelines for posting in this Scientific Section and try to adhere to them. I assume that these guidelines no longer apply now?

Lastly, you are being lazy and trying to cast odubts on the research when you state:

I'm also unsure whether the research is peer reviewed?? I can't see that it is??

All you need to do is Google a publication to determine if it is peer reviewed or not. Anybody reading my posts on these topics is always provided with ample information/links to see what I have seen or to answer some of the simpler easy to answer ones like this.
Guidelines for Reviewers of ASM Journals This is from the American Society from Microbiology Journals site which published the 3 papers mentioned in this thread by Hovanec et al.


OldMan47-

I have not read through the references you gave but am the first to agree that Tim Havonec at one time had the definitive research into the identity of the predominant AOB and NOB species. What in the world does that have to do with anything you are supporting today?

First, whose research has supplanted Dr. Hovanec et al. (you too seem to forget there are other names on everyone of the studies in question). Surely if the research has been supplanted by newer research you should be able to link us all to this research? Secondly, how dare you ask what this has to do with what I am supporting today in view of the fact that you issued this challenge to me in the thread from which this one was spawned:

Do you have any scientific evidence at all that the Safe Start contains the right bacteria or any live bacteria TTA

That is the whole point of this thread. If you want to bottle bacteria which perform nitrification in aquariums, don't you think that the very fist step would be to identify the bacteria? There are two reasonsfor this thread. The first is that Ianho has called into question the validity of Dr. Hovanec's research because the ultimate result was his bacteria in a bottle. Ianho claims this fact invalidates the research works of Dr. Hovanec et al. The second is because you issued the above challenge and was the reason I added your name as part of my challenge.

This thread, and the links it contains, is how I know the bacteria in TSS are the right ones because they are basically the same ones as in One and Only since Hovanec is responsible for both. If you doubt this then do the homework yourself to find out the history of the patents, the research companies and how Tetra ended up being the inheritor of Marineland and Aquaria Inc.'s labs and shares in the right to use the bacteria which are patented. It is also the reason why Dr. Hovanec has his own company. His labs are the old Aquaria Inc./Marineland facilities. I have found this information, so why can't you? Perhaps it is a good idea to check the facts first and then to post instead of the other way around?

In order to come to any reasonable opinion if the bacteria in the bottle are the right kind, you should know how and why they are in the bottle. The next step is to know whether non-sporulating bacteria can survive in there. I have provided ample research links (in other posts as well as this reply) totally unconnected to Hovanec et al. that indeed they can. And once we work through phase one here, accurately Identifying the bacteria in aquariums that do the nitrification, I will organize a thread which deals with the preservation of the bacteria with all the relevant scientific research. It too will be link rich and scientific in nature.

Moreover, you attribute a statement to me which is not true:

On the other hand you seem to be supporting nostrums like Cycle and Dr Tim's One and Only which are not refrigerated

Nowhere on this site nor anywhere else on the net can you find me ever having said I believe Cycle does anything. However, I have said in various threads here that I have personally used One and Only twice and it worked as advertised for me. But his is not science.

 

[lloyd]

i happen to find this topic interesting and i really didnt want to post on this topic because i have nothing scientific to add.

if people like tom bae and mint took the time to grow up and read the rules they will see that they have broke many of them so im asking if they'll just go away and let the topic run its course as its obvously above them and there just annoying the people reading it.

here is a link to the rules if just incase you cannot locate them

http/www.fishforums.net/index.php?/topic/205832-guidelines-for-posting-to-the-science-forum/

i know posting this technically has broke the rules but i have the interests of science in mind

 

[atlas101]

@twotankadmin, I have just had a quick read through that literature review type paper you posted and the part where they discuss the viability of starved bacteria in a refrigerated state seems to be drawn from two papers:

Wang X, Zhang H, Yang F, Wang Y, Gao, M. (2008) Long-term storage and subsequent reactivation of aerobic granules. Bioresour Technol 99:8304-8309

and

Yilmaz G, Lemaire R, Keller J, Yuan, Z. (2007) Effectiveness of an alternating aerobic, anoxic/anaerobic strategy for maintaining biomass activity of bnr sludge during long-term starvation. Water Res 41:2590-2598

I was just wondering if you happen to know of any free versions of those papers, as I would like to have a read myself but dont fancy paying between $30-$50 a piece to read them!

 

[OldMan47]

I find it interesting that we are still discussing bacteria when it is gradually being shown that archaea, most often called AOA in the literature I have found, are the dominant ammonia processors, not bacteria. The effective populations of AOA are usually at over 10 times the population of AOB in most of these studies. I have been searching this evening for good references and am finding it difficult since most research has actually been done on marine environments, not freshwater ones. However the articles posted here, here and here have plenty of clues to help us find the most recent research findings. In the meantime I am engaged in trying to follow some of those clues to get more definitive, or at least supportive, research results. As stated in many places, research that has not been confirmed by independent research is at the least suspect unless no attempt to replicate it has been made.

Once we truly understand what is going on, we may go in a whole new direction, but without independent analysis of "bacterial supplements" I will continue to doubt their value. Has any scientific evidence been presented yet that supports commercially purchased TSS or Dr Tims that was not conducted by the respective companies or an industry expected to benefit from them being found acceptable? I could easily bottle my filter contents, remove them from the bottle 5 minutes later and cycle a filter in less than a week. It is equivalent to what I do all the time now that I have well established filters to use as a cloning source. Going to several local shops, not chains that buy in bulk, and buying the product off the shelf without sorting out just the most recently bottled sample would at least be a step in the right direction, if and only if it was not done by the respective company.

 

[TwoTankAmin]

The evidence you speak of is far from conclusive. I have seen and read more studies than the three you linked. I had already read two of them. You are correct that it seems to apply only to sw. Moreover there is a lot of debates as to what the respective roles of AOA vs AOB may be. One of the confounding problems is that since the initially discovery of AOA in a public salt water aqaurium in 2005 there have been numerous studies but none to date has found any nitrite oxidizing archaea. Even when AOA are found in numbers it appears that it is NOB that handle the nitrite oxidation.

Another issue to be resolved is how the AOA process ammonia vs the AOB. Whereas the AOB rely on iron as an essential part of the process, the AOA utilize copper. In the open ocean up from the depths the earth spews some pretty nasty stuff. Copper in its toxic form is one of them. It has been speculated that the AOA may play a role in removing the copper on the way up from the depths. Algae wont grow until copper levels are low and any remaining copper has been converted to its less toxic form. This opens the entire Archaea issue to a whole other area of study yet to be done. It may explain why the AOA have turned up in public aquarium's sw and not fw tanks. Fw life forms are much more senstive than their sw counterparts to copper and in general natural fresh water copper levels are lower than those for sea water. These lower levels may prevent AOA from being in fw tanks.
Here is the research on this Nitrosopumilus maritimus genome reveals unique mechanisms for nitrification and autotrophy in globally distributed marine crenarchaea (Much of this study left me in the dust, it is mostly way too technical for me.)

Moreover, the iron the AOB require is highly variable in different areas of the ocean which would favor the copper using AOA over the iron using AOB in open oceans. In aquariums everything is pretty evenly distributed throught the water. Finally, when you are talking about the open ocean, there is no hard surface for bacteria to attach to. In aquariums the AOB are on the hard surfaces inside biofilms. So I know how the nitrifying bacteria in tanks are still mostly there after a 50% water change. But if some portion of the AOA are free swimmers, what happens to them after a 50% water change?

Research also indicates that the AOA can be more photosensitive than the AOB. This may explain why the initial discovery was on a rock from the bottom of a public aquarium sw tank. (A tank which gets its water from the open sea.) It appears as if in most tanks the light levels could inhibit the AOA. Sorry its only the abstract, but Differential photoinhibition of bacterial and archaeal ammonia oxidation

Here is a paper which studied the ammonia oxidizing communities in a marine aquaculture system. This is much closer to an aquarium than soil studies, open ocean studies, estuary and coastal water studies etc. They describe the AOA contribution to nitrification the bio-filtration system is "negligible". And this time it is a full study that one can read Nitrosomonas Nm143-like ammonia oxidizers and Nitrospira marina-like nitrite oxidizers dominate the nitrifier community in a marine aquaculture biofilm

Finally, another interesting fact about the AOA. They have the ability to survive and thrive at lower ammonia levels than any of the ammonia oxidizers. The AOB are unable to establish/thrive at these low levels. At higher levels the AOB tend to out compete the AOA. Finally, most of the AOA research which refers to aquariums is not about fish tanks but about large public aquariums. Many of these are on the coast and get their water from the the sea. Most sw fish tanks mix up their water, it doesn't come from the sea.

So now that we can see when it comes to the whole Archaea issue, especially as it relates to fish tanks, that it is far from

"gradually being shown that archaea, most often called AOA in the literature I have found, are the dominant ammonia processors, not bacteria."

I suggest you might want to read a bit more on this topic. But until then lets stay on the topic of this thread, which you are still artfully dodging. The identification of the predominent AOB and NOB in home aquariums as the initial step in being able to create a bottled bacteria. If you want to start a separate thread on AOA, I would be happy to participate.

As to this:

Has any scientific evidence been presented yet that supports commercially purchased TSS or Dr Tims that was not conducted by the respective companies or an industry expected to benefit from them being found acceptable?

We have had this discussion over and over. There is no reason for somebody to do such reseach and even less for somebody to fund it. But in terms "the expected to benefit" group. I posted about several major public aquariums that have used his product to fast cycle huge tanks in a hurry and then placed decent fish loads in the tanks. These aquariums did not all do this on the same day or month. So lets assume for a minute that you are correct and One and Only can not and does not work. So they poured this stuff into a huge tank, then they dropped in a good stocking of inhabitants (the dolphins in the Georgia Aquarium are not fish they are mammals, but they make ammonia ). What should have happened on these tanks. Since they are well stocked and have no biofiltration, what happened?

And don't try and say they did massive water changes on 1/2 to 2 million gallon tanks for weeks on end. Especially if they don't have access to sea water as in Atlanta, Georgia; Tempe, Arizona or London, England. Sea Life at Legoland in California would have had that choice. But what makes you think three different large public aquariums all run by the same corporation would have one after the other all used a product? Surely they would have known it did not work right after the first one used it? Does that make any sense to anybody??? Plus the Georgia Aquarium would surely have gotten wind of all this too so why did they use it as well? The financial losses, the bad publicity all would have been a big deal. Many millions of dollars wasted, many millions in lost revenue. And yet nobody angry enough about it all to publicly skewer Dr. Hovanec or at least to sue his butt off?

I would suggest this anecdotal evidence very strongly suggests the product must work, at least in salt water. And if it works in salt water, doesn't that get one pretty close to being able to believe the right bacteria might also be in the fw product and could also be alive?

And to loop back to the AOA, if none of the three land locked Aquariums above uses sea water in their systems, exactly how do you think AOA would have gotten into those systems and then reproduced sufficiently to be the dominant ammonia oxidizers?

 

[OldMan47]

I can agree with your anecdotal evidence that the additives used by the commercial aquaria were effective to some degree, what in the world does that have to do with 8 oz samples found on the shelf of my local fish shop? These people order directly from large corporations and can expect far superior responses than you or I would get buying the stuff after it had been on the LFS shelf for 6 months. I am afraid that we are comparing apples and oranges. Go back to my last post. Do we have any real world research that draws on available "off the shelf" bacteria in a bottle that supports its effectiveness? If we have that, I would be thrilled to accede to your prejudice that the stuff must work because the company days it does.

 

[ainsy]

I completely agree that that is apples to oranges. It also strikes me that there is no comparison between a 1 million gallon aquarium and the average sized one found at home - there is obviously a dilution difference there. The fish load to volume ratio of a HUGE public aquarium can't compare to that of the average home aquarium.

I am sceptical of any research conducted by someone who stands to gain financially from the success of a product. It isn't that long ago that cigarettes were being advertised as a cure for all kinds of illness.....

 

[ian]

Now, you and some other folks asked to see any other evidence that bacteria can survive in a bottle. Here is a 21 page review on this topic. It is the work of a consortium of 7 European companies in businesses related to aquaculture etc. This paper Section 2 - Methods for preserving bacterial cultures. Section 3- Storage of nitrifying bacteria. Section 4- Factors affecting the survival of nitrifying bacteria during storage (4.1 Ammonia-oxidizing bacteria - 4.1 Nitrite-oxidizing bacteria). This paper includes references to some 87 other studies, one of which is Hovanec et al. This can be opened in Microsoft word. It clearly shows that there are a variety of working methods for storing nitrifyiers as well as other bacteria.
http/adapond.eii.e...eliverable.docx

^^now this is probably the most interesting article in this thread TTA. This is getting more to the point of what we are getting at with regards of shelve lifes etc etc. I mean this when i say thanks for posting. I'm struggling to find the way Dr Tims's and Tetras bottles preserve, do you know?

Ianho, this has to be some conspiracy spanning multiple decades, assorted universities and research organizations and literally many 100s of scientists. All designed to make Dr. Hovanec a few dollars from selling his bacteria.

again, if you bothered to read my replies, you know why you must always approach research this way. It's not conspiracy, its the way academia teach's you.

I did find an old thread of yours, and you did state that you had no formal training at reading and critiquing research, which does show in the way you post and link things, it's really not very user friendly.

I also found a thread of yours from not so long back with your emails to Dr Tim regarding bottled bacs and his explanation to you was poor...

this was his explanation to you when you asked 'why don't bacs die in the bottles?'

Again this is semantics. The cell may be alive but it is not useful to use in the sense that it cannot rapidly (in hours/days) convert ammonia to nitrite. Bacteria in a bottle can be revived and even an old bottle of bacteria is better than no bacteria (as long as they were the right bacteria in the first place). But people expect things in different time periods.

The reason for the "old wives tale" that bacteria in a bottle don't work is due more to the fact that the bacteria in the bottle were the WRONG bacteria in the first place so they didn't even work before placed in the bottle.

But for the cells to stay active they need to be fed and this cannot happen in a bottle. So once placed in the bottle the bacteria activity slowly fades - they are not dead but they are not real active hope that makes sense

^^this is a poor poor explanation and i'm unsure why you didn't rip it apart!!

again thanks for the word doc as it is the only link that has any bearing on what we are talking about.

 

[shrimply]

An interesting paper that may contribute if you can get your head round it( going over mine right now, scrambled from the exam this morning)

http/www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0023281

Looks at both Archea and Bacteria and Bottled products, also uses smaller type Freshwater aquaria so may be more valid.

Just a quick note on the comment regarding the "rules", I actually think they are a bit ridiculous, are you trying to tell me all scientific discussion takes the form of a scientific paper. Scientists bounce emails back and forward, suggest ideas, agree, disagree, change their mind, argue, exchange interesting papers. That's what scientific discussion is and what it should be. Free exchange of ideas that leads to testable hypothesis.

 

[TwoTankAmin]

That Archaea paper should be looked at with a lot of skepticism. I can tear it apart on several fronts. But lets start with a few quick easy issues. Read the introduction and you will find very early on the reference all 3 Hovanec et al. papers. Just to remind you, paper one in 1996 discovered the aquarium nitrifyers were not what they expected to find, which were the traditional ones. Because the ones everyone expected to find were not there, they concluded the actual bacteria doing the work were not yet known. The second paper from 98 was the one in which the nitrite oxidizers were identified and the third paper in 2001 identified the ammonia oxidizers. So the authors of the study reference the paper entitled "Identification of Bacteria Responsible for Ammonia Oxidation in Freshwater Aquaria." Now here is what the Archaea paper says:

Based on the ubiquity and high abundance of AOA in natural environments, the inability of Hovanec and DeLong (1996) to detect AOB in freshwater aquaria, and the isolation of the first ammonia-oxidizing archaeon from aquarium substrate [10], we hypothesized that AOA dominate freshwater aquarium biofilters and play an important role in aquarium nitrification.

So they knew Hovanec et al had identified both the ammonia and nitrite oxidizers in fw and sw in aquaruims and yet they make that statement. That set off my first alarm. Their hypothesis was based on ignoring the info they referenced. I would suggest they were biased from the sart.

In the introduction they also reference two other studies, both of which I have read and which you can find in their full form
Low Temperature Decreases the Phylogenetic Diversity of Ammonia-Oxidizing Archaea and Bacteria in Aquarium Biofiltration Systems[down-pointing small open triangle]
Nitrosomonas Nm143-like ammonia oxidizers and Nitrospira marina-like nitrite oxidizers dominate the nitrifier community in a marine aquaculture biofilm
They fail to mention two things in their introduction, which also references the initial study in 2005 about the discovery of the first confirmed strain of AOA. First they simply refer to the discovery taking place in an aquarium. What isn't mentioned is these are not fish tanks, they are public aquariums on the coastlines (on in California and the other in Japan). The odds are very high both get their salt water from the sea. I thing, but am not sure the California one said they do but I could not find that info regarding the one in Japan, but is seems highly likely they do. So is any surprise that the Archaea which are massively present in the oceans would end up in huge tanks filled from the sea? The cultured used in the aquaculture study came from a fish farm in Israel and it makes its saltwater rather than taking it from the sea.

The next thing to look at is their information regarding the 35 tanks they used in their study. A number of the fw and sw tanks sampled came from the same two retail fish store's display tanks. Exactly how hard is it to imagine the potential for cross contamination between them? This is far from a controlled scientific source. And this brings me to another issue. You can see the parameters and other related data in Figure 1. It shows tank by tank information for gene copies for bacteria (none of which is ever identified by name anywhere in the study), Archaea and for the amo for each. it also lists info on tank parameters etc. They used sophisticated equipment to test for ammonia and for pH, but they state they performed the tests for nitrite, nitrate, hardness and alkalinity using Jungle Test Strips. Hmmmm.

Finally, for now, take a look at Figure 1 on the tank info and if you work at it, you will see some strange inconsistencies.. You will also see they report 4 tanks with ammonia between .33 and .56 ppm. 7 tanks have nitrite readings of .5 to 3 ppm. 1 tank had been dosed with antibiotics sometime in the preceding 6 months. Not counting any overlaps, there are 9/38 tanks which have questionable parameters in terms of what they may or may nor represent or what effect any of this might have had on results.

This is just a start on what I see as some major issues in the report that that caused me to question it. Considering how hot a topic Archaea have become in the discussions of nitrification, why is it that this study has only been cited once since it was published last August? Also take a look at the level of detail this study provides on its methods vs those in any of the 3 Hovanec studies. This one seems to be fairly short on information by comparison.

Ianho, yes, its fairy obvious if one researches how non-sporulating bacteria survive hard times. There is a wealth of research on this topic. This research has nothing to do with Hovanec et al. But again all of this is again off topic. The issue to be resolved here is are the bacteria that go into the bottle the proper ones for our tanks or are they not. If they are, then we can move on to discussing how they can last when in the bottle. If they are not the right ones, it doesn't much matter even if they could survive for 1,000 years exposed on the moon does it?

I am skeptical of any research conducted by someone who stands to gain financially from the success of a product.

Even if that research was performed years before the product was developed? Hovanec et al. (and again the et al. people had/have no financial interest in the products save perhaps for Ms. Phalen) researched what bacteria was doing the work. And once they found that answer they, bottled it? Not the other way around.

Oldman you are some piece of work. Partially worked? I was unaware that bacteria partially work. So does that mean they work 1/2 days? They only nitrify so much and then go home? If they partially worked, the tank inhabitants would have been in trouble. How much bio-load is produced by an adult dolphin and what it eats and what fish etc. they surround it the with. Big tanks are for big size and big numbers of fish. They produce big bioloads. Nitrification issues in tanks are all about ppm not the amount of gallons of water.

More later, life has become overly busy with other stuff so I will post more as time permits.