An Open Challenge To Ianho -

[Dieses Madchen]

I know this isn't quite the discussion but I wanted to share as it contributes to the bottled bacteria discussion. I cultured a bottle of Biozyme, which is supposed to be used as an aquarium starter. The first pic was taken of the plate that was incubated at 30C.

This plate was incubated the same amount of time as the other, only the temp was 37C

Even thouhg I did not have time to get a definitive ID on the colonies that grew, I can tell based on the plate it grew on as well as the morphology (gram negative bacilli) that it is a nitrifyer. I do not know if it is the same as what grows naturally in the filter. I would like to, over the next few months, do more studies on these products to find out more about them and how they compare to what naturally occurs. I do know that the bacteria in a bottle seem to act the same as natural filter bacteria in the conditions it grows, higher temp equals higher growth rates and what not. When I get more time to do some more research I will post more. But I hope this is of some interest.

 

[atlas101]

The bacteria are in continuous production, they are not just whipped up. They are grown under controlled conditions. I have read descriptions of how it is done and what parameters are maintained.

I folks in general were a bit more open minded about Dr. Hovanec's background and what he does beyond this one product, they would find there is a wealth of information available from him. You can learn about the biofilms in which the bacteria live, why bio-wheels are good hosts for them. You can learn things about cycling and bacteria that have nothing at all to do with bottled bacteria. It is a mistake to merely dismiss any information that he provides in this area simply because he happens to sell a product.

The fact that the bacteria live in a biofilm has nothing to do with a product, the fact that elevated ammonia and/or nitrite levels inhibit nitrifying or kill bacteria has nothing to do with bottled bacteria. The fact that nitrifying bacteria do have strategies for surviving times of little or no food or oxygen exists independently of the existence of bacteria in a bottle products, though it does explain why they can last in a bottle.

One can do a Google Scholar search for Nitrospira and get back some 3600+ studies and books. You will read lines like this:

Nitrospira are barely studied and mostly uncultured nitrite-oxidizing bacteria, which are, according to molecular data, among the most diverse and widespread nitrifiers in natural ecosystems and biological wastewater treatment.

From A Nitrospira metagenome illuminates the physiology and evolution of globally important nitrite-oxidizing bacteria (incidentally this 2010 study is edited by one of the et als. of the Hovanec et als. I often mention.)
or

Chemolithoautotrophic nitrite oxidizers of the genus Nitrospira are a monophyletic but diverse group of organisms, are widely distributed in many natural habitats, and play a key role in nitrogen elimination during biological wastewater treatment. Phylogenetic analyses of cloned 16S rRNA genes and fluorescence in situ hybridization with newly developed rRNA-targeted oligonucleotide probes revealed coexistence of uncultured members of sublineages I and II of the genus Nitrospira in biofilm and activated sludge samples taken from nitrifying wastewater treatment plants.

From Nitrite concentration influences the population structure of Nitrospira-like bacteria

So atlas101 you don't need dirt- you can use waste water sludge. But I think you would much prefer to have the bacteria without all the other stuff.

I dont think people specifically doubt this Dr. Hovanec's research or the science behind his papers, given there is alot of collaboration and other authors who support the identification of the nitrospira bacteria. I think the main problem people have is once you have this bacteria rich liquid, how long can it last? in the literature review you posted earlier on in this thread, they came to the conclusion that these bacteria can last upto several months but the two papers they drew this conclusion from were not really the same anaerobic conditions that would be present in a bottle, in one study they used an aeration period of 15 mins per 6 hours and in the other the bacteria were stored on something called aerobic granules, not in liquid form in a bottle.

I think an experiment directly testing and measuring the survival rate over a given period of time and reactivation period of the bacteria in a liquid bottled form would be worth its weight in gold to the companies producing these products it would seriously sway the doubters opinions and probably boost sales of these products aswell. If such research exists already I would love to see it, it would convince me. There is alot of research out there on how many kinds of bacteria can survive many adverse situations and periods of starvation but there are always slight differences to the research, definitive research using the actual bottled bacteria products themselves would help the cause no end.

I have never personally used one of these products, but I think I am going to try it next time I have to cycle a filter, I have always felt that in theory this type of thing should work, the only thing that has put me off is the large number of people for whom these products didnt work, reading across many forums these products only seem to work around 50% of the time. Maybe these people just did something wrong, I dont know but at least you have convinced me to give it ago myself.

 

[TwoTankAmin]

DM--> Biozyme is a revolutionary product utilizing dried culters of hetrophic bacteria and enzymes fixed on bran.
http/theaquariumwiki.com/Biozyme

Here is a hoot. If you use One and Only to do a fishless cycle and you follow the instructions on DrTims site, you should be able to have a tank fully cycled in a week give or take 2 days.

All you need need is his bottled bacteria and his ammonium chloride (normally included w/ the bacteria), a tank with some substrate, a filter that also aerates, a heater and a pH over 7.0 (and higher is better up to about 8.3ish).

To know if it really works or not takes about a week. But you must do it properly, i.e. according to the instructions.

And you are correct, the major cause of failure using Dr Tims or Tetras Safe start is user error. it is impossible to put full information on the bottle. So one should use the respective sites to find in depth information and directions. DrTims label should direct you to his site. I know the ammonium chloride one does as I still have 2. I threw out the empty bacteria bottle. You can use the ammonium chloride to cycle with seed material from other tanks too.

Many people throw in the product to save a cycle out of control and have ammonia and or nitrite levels over 5 ppm. That usually insures it wont work. Many people throw it into tanks over dosed with ammonia binding products which is also not good for the bacteria either and can inhibit them.

 

[Dieses Madchen]

DM--> Biozyme is a revolutionary product utilizing dried culters of hetrophic bacteria and enzymes fixed on bran.
http/theaquariumwiki.com/Biozyme

Most bacteria in a bottle is a hetrophic nitrifyer.

 

[TwoTankAmin]

And to paraphrase Will S.- "Therein lies the rub
For in that bottle of bacteria we know not what nitrifyers may come...."

Please circle back to the original post.........

 

[Dieses Madchen]

And to clarify, i was just showing some of the research i have done on my own with these insta-cycle products, whether its liquid bottle or freeze-dried. Also stated that it was quite the discussion. I just know a common question is "does it actually have bactera that is live."

Sorry that my comment didn't relate to the original post. I believe I stated that it didn't. Dont have to be so condescending

 

[TwoTankAmin]

DM- you misunderstood my point. I was trying to point out why there is a difference between One and Only or Tetra Safe Start and most other products. Your post was great because it showed that the bottles do contain culturable and/or live bacteria. But it is not the heterotrophs that we want, but rather the autotrophs identified in the Hovanec at al. studies. Which is why I said lease circle back to my original post."

Your post probably did more than almost any on this thread to illustrate my point. (It definitely relates to the topic at hand as well.) And where it leads, imo is back to the studies I referenced at the outset.

There are a lot of products out there that make all sorts of claims. Very few will tell you exactly what is in the bottles. OaO and TSS both tell you. Even the Biozyme description only says they are heterotrophic but not which ones. And this was what I was trying to point out in my paraphrase of the bard. I wasn't talking to you, it was meant for readers in general.

 

[Dieses Madchen]

DM- you misunderstood my point. I was trying to point out why there is a difference between One and Only or Tetra Safe Start and most other products. Your post was great because it showed that the bottles do contain culturable and/or live bacteria. But it is not the heterotrophs that we want, but rather the autotrophs identified in the Hovanec at al. studies. Which is why I said lease circle back to my original post."

Your post probably did more than almost any on this thread to illustrate my point. (It definitely relates to the topic at hand as well.) And where it leads, imo is back to the studies I referenced at the outset.

There are a lot of products out there that make all sorts of claims. Very few will tell you exactly what is in the bottles. OaO and TSS both tell you. Even the Biozyme description only says they are heterotrophic but not which ones. And this was what I was trying to point out in my paraphrase of the bard. I wasn't talking to you, it was meant for readers in general.

Thank you for clearing that up

 

[Prime Ordeal]

I know this isn't quite the discussion but I wanted to share as it contributes to the bottled bacteria discussion. I cultured a bottle of Biozyme, which is supposed to be used as an aquarium starter. The first pic was taken of the plate that was incubated at 30C.

This plate was incubated the same amount of time as the other, only the temp was 37C

Even thouhg I did not have time to get a definitive ID on the colonies that grew, I can tell based on the plate it grew on as well as the morphology (gram negative bacilli) that it is a nitrifyer. I do not know if it is the same as what grows naturally in the filter. I would like to, over the next few months, do more studies on these products to find out more about them and how they compare to what naturally occurs. I do know that the bacteria in a bottle seem to act the same as natural filter bacteria in the conditions it grows, higher temp equals higher growth rates and what not. When I get more time to do some more research I will post more. But I hope this is of some interest.

Dieses Madchen, we've covered this one already. Biozyme contains ONLY heterotrophic bacteria and so your plates are meaningless.

I'll try to find the post where you freaked out the last time I challenged you on this.

DM--> Biozyme is a revolutionary product utilizing dried culters of hetrophic bacteria and enzymes fixed on bran.
http/theaquariumwiki.com/Biozyme

Most bacteria in a bottle is a hetrophic nitrifyer.

NO.

I must also admit that there MAY be something in this product (swallows pride lol), but i would like to try it myself.

Pretty much says it all.

 

[ian]

Say's all what Prime?

TTA has presented some decent evidence in this thread, some not so good...but enough for me to query the product a little more.

 

[TwoTankAmin]

Ianho- it is very easy to check the product. On DrTim;'s site are directions for doing a fishless cycle using his One and Only. It says is will take between 5 and 7 days to have a tank fully cycled. Of course you must follow his instructions there for doing the cycle. One can't change the methods, then get different results and then claim it doesn't work.

He used to, and may still, include a bottle of ammonium chloride designed to give 2 ppm of ammonia when dosed at one drop/gallon.

You will note you do not dose much ammonia and then you only dose twice with dose 2 being 1/2 the amount of the initial dose. The third dose it to "prove" the cycle done.

 

[Dieses Madchen]

I'm not hear to argue with you prime ordeal. Me and the OP have had discussion off of this thread about the plates and the pictures were more so directed to him that anyone else. Whether or not your think they are meaningless was not really what I was after. But thank you. This WAS a civilized thread

 

[FishFanatic04]

We need an experiment. People from different countries starting on the same day and doing the exact same things to their tanks. It would be more informative if it were done by hobbyists, though perhaps less scientific. I can continue reading the linked articles until my eyes cross but as Ianho said: I would like to see for myself.

I have a scrubbed out tank and fresh filter sitting around with nothing to do.

That will also leave room to weed out user error.

 

[Dieses Madchen]

I started to do that a while back, Fishyfriends. Never had time to finish it...

I think its a great idea.

 

[TwoTankAmin]

So it seems to me now that the issue is not one of what to put into the bottle, but rather can they really live for 6-12 months and still function well in fairly short order or not. A while back I came across a description of the bio-farms Dr. Hovanec's company uses to produce the bacteria, unfortunately I did not bookmark it and have been unable to find it again. It involved pure water, a carbonate to buffer and to provide inorganic co2 and hold the pH up and circulation for O. Temp. is kept optimal and the bacteria are kept well fed to maximize reproduction. The idea is based on the research which indicates not only do different bacterias have different lasting abilities, but also that the condition they are in when they go "dormant" has a big effect on the lasting times as well.

Before moving on to the next thread on how long do they last, I would like to propose the following:

I have two empty 10 gallon tanks. I am prepared to purchase two more. I have sufficient air pumps to drive a sponge filter for each. I have loads of used small inert gravel I can bleach to sterilize and which should neither contribute to nor impede a cycle in any way beyond providing a surface on which the bacteria can attach. I will have lids for all the tanks and will put heaters into each as well. Moreover, I have an RO/DI unit, so I will fill the tanks with water as pure as possible. Since we have our own private well, I have never used dechlor, and so will not here. I will buffer the water using sodium bicarbonate (NaHCO[sub]3[/sub])*. I have a digital TDS/Temp. tester and will get similar brand thermometers to go inside each tank. * To raise KH without raising the GH, add sodium bicarbonate (NaHCO[sub]3[/sub]), commonly known as baking soda. 1/2 teaspoon per 100 Liters raises the KH by about 1 dH. Sodium bicarbonate drives the pH towards an equilibrium value of about 8.2.

I will use multiple brand test kits. To measure ammonia, nitrite and nitrate I will use both API and Seachem kits. To measure KH and pH I will use API kits. I will use the following parameter guidelines and will work to keep each tank as close as possible within these ranges: Temp. 82-84F/27.77-28.88C pH 8.2-8.3 KH 3-4dg or above Initial ammonia dose to 2 ppm TAN. 1. I will purchase four 2 ounce (29.57 ml) bottles (sufficient to cycle one 30 gal tank) of Dr Tim's One and Only from four different sources including one from Dr. Hovanec and the rest from:
http/www.drsfoster.../ps/c/3578/3975
PetStore.com
Ebay Seller
Since I can't control the dates on any vendor's bottle, if any are over 6 months, I am willing to use them but not to be held to the below defined schedule of achieving a full cycle in about 5-7 days. However, using an out of date bottle can also provide useful information so I feel all bottles should be used regardless. Further, since I have purchased from DrTim's site in the past and have corresponded with him, I will not order the bottle from him myself. I will have a friend in Danbury, CT order it and I will get it from her (she is about a 30 minute drive from me). This should give us an idea what the average person trying this product might buy.

2. I will sterilize the 2 used tanks and lids and sponge and airline parts (not including the new sponges themselves) in a strong chlorine bleach solution. Say 20-25%? and let them be rinsed well then and air dried. I wiil thoroughly rinse new tanks and equipment.

3. I will first rinse the gravel and put a thin layer (.5-1.0 in./1.3-2.5 cm) into the tanks to cover the bottom glass. I will then add the RO/DI water, measuring it to know the exact amount and endeavor to use the same volume in each tank. I will add the filters and heaters and turn them on.

4. When the water comes up to temp. I will test all the parameters and then I will add the buffer to achieve the pre-dermined levels as measured by pH and kH.

5. When all the parameters are at acceptable levels, I will add the ammonia to dose the tank to 2 ppm. I would prefer to do this using the ammonium chloride available from Dr. Tim's. It should provide a known strength and should produce 2ppm/gallon of TAN when dosed at the rate of 1 drop/gallon. His site suggests dosing between 1 and 3 ppm when doing a fishless cycle. Since a 10 gal tanks actually hold less than 10 gallons, dosing to 3 ppm will result in a greater overage potential than 2 ppm as fractional drops are not possible.
6. I will wait 30 minutes and test the ammonia. If it is close to reading 2 ppm, I will begin the adding the bacteria. If it is not close I will add more ammonia or change enough water to bring it into range. I will add between as close to 10ml (1/3 of a bottle for a 30 gal and the smallest size bottle available) of Dr. Tims One and Only. I will shake the bottle well before measuring the ml. 10ml is the minimum needed for a 10 gallon tank. Each tank will be dosed from a different bottle. I will then continue to follow the method as described by Dr. Hovanec on his site[sup]1[/sup] for doing a fishless cycle using his product. He suggests an initial dose of ammonia between 1 and 3 ppm. I chose 2+ as it makes the whole thing simpler with drops that - 2ppm/gal. My initial dose will be 10 drops. (I am assuming the tanks will not hold a full 10 gallons so the dosing will amount to a tad over 2 ppm.)

7. The next day as close to 24 hours later as is possible, I will dose 5 drops of ammonium chloride- 50% of the original dose from the previous day.

8. No more ammonia will be dosed until day 5. At this time I will dose ammonia equivalent to 2 ppm (10 drops).

9. On day 6 I will test for ammonia, nitrite and nitrate 24 hours after the day 5 dosing of 2 ppm (1- drops). If these read 0/0/and show nitrates, we will assume the tank is cycled. If there is over 0.5 ppm of either ammonia or nitrite I will agree the product has failed. If there is a little bit of ammonia/nitrite, 0.5 or less of either or both, then I will dose 2 ppm (10 drops) again and measure in 24-48 hours. If tanks are not at 0/0/nitrates risen, then I will agree the product has failed to perform as advertised.
[sup]1[/sup] http/www.drtimsaqu...ng/how-to-start Scroll down to the section entitled "Using DrTim's Aquatics One and Only Live Bacteria"
Since I can not control the age of the bottles as noted above. Any tank that proceeded using an out of date bottle will not count as a failure if the tank doesn't cycle on schedule. It will count if it does meet the cycled criteria in the specified time frame. I will keep trying to get any out of date tank fully cycled to determine if it goes any faster than a non-seeded fishless cycle, that is, fewer than 30-35 days or so.

To allow for the possibility that one of the three outside vendors might have killed the bacteria off due to mishandling, there should be a 1 tank failure permitted without prejudicing the study.

Feel free to suggest modifications to the above (except for the actual ammonia dosing regimen) as long as it is not an absurd condition or requirement.
[sup]1[/sup] http/www.drtimsaqu...ng/how-to-start Scroll down to the section entitled "Using DrTim's Aquatics One and Only Live Bacteria"

FYI- My tap water has a pH of 7 to 7.2. It has a GH of about 4-5dg and a KH of about 3-4dg and the TDS can range from a low of 58 ppm to a high of 85 ppm depending on how recently it has rained and how much it rained. Running it through the RO/DI should result in pH 7.0 and both GH and KH between 0 and 2 ppm which might read 1 dg on the test kits but which, in actuality, would be virtually nil.

Of course I would post all the ongoing results and pictures. The nice thing about all this is it should only take about a week once it starts.
What do folks think about this? Any suggested changes, with an explanation for making them will be considered.