EMeyer

This forum needs a "mind blown" reaction. Cause, /mind blown My dry rock + bacteria in a bottle experimental tanks have very low scores, despite strong nitrification My gut says there is more to your success than dry rock + Dr Tims, but can't presume to know what!

One person can... Share away, by all means. My tendency is to share everything. My wife and various employers have informed me thats not always appropriate, but what do they know?

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Thanks, as I prepare descriptions of the results I will reach out as needed to ask about permission to share. As a default company policy it seems appropriate to preserve privacy of everyone's data, although when people are happy to share data that always helps! There are a lot of questions we can ask anonymously; e.g. many of the samples came from tanks with cyanobacteria, we can ask about signature of cyano problems without disclosing anyone's specific data. But there are a few samples that may be useful examples to discuss so I will reach out to the owners when I get to that step in writing. I greatly appreciate peoples' willingness to freely share data, that is (supposed to be ) how science progresses!

Ladies and Gentlemen, Your reports are ready - to view your own report(s), please see this link (you'll need to log in to view your report) https://aquabiomics.com/view-reports/ I will do the "your data are ready" individual emails through my website as soon as I set that up. Its just that analyzing these data and writing programs to generate these reports has been interesting and all-consuming. Web development stuff is far less interesting although necessary too. I will make a series of posts in the near future describing these reports and what we've learned. Specifically A guide to interpreting your report What have we learned about the microbiome of a typical reef tank? What have we learned about various methods for establishing the proper microbiome in a reef tank? For now I will just say this -- please remember that for this first batch of samples, the balance and diversity scores are based on comparing you all with each other. (None of my tanks were included in the "typical reef tank microbiome" database). Diversity is pretty easy to understand. Its the number of microbial types in your tank. I've also expressed this as a percentile (the diversity score) comparing your tank with everyone elses (1=most diverse tank, 0=least). Balance scores are based on comparing the abundance of different bacterial types. If your balance score is high, that means your tank's microbiome is like most other tanks. If your score is low, it means your tank is different. It is too early to say whether that is good or bad -- just different from the typical pattern. Maybe your tank is right and the typical tank is wrong! I think it will be important to consider the balance scores in the context of the health of your tank. And as we continue to build a database of healthy tanks these scores will become more diagnostic. There is some interesting stuff here. I have been too busy with coding to write up scientific stories about it, but thats coming next... for now, please enjoy your data and don't hesitate to ask questions especially if you have any difficulties accessing them... I will appreciate any and all feedback! -Eli

I've got one thats drilled in the bottom on one end. If that works for you, I'm close to Corvallis.

Hi all, What a summer. I hope yours has been less hectic than mine. But despite having to invent several non trivial things along the way, and everything taking longer than it should (the sequencing facility took over 1 month to complete a 48 hour job!)... I have very good news. Every sample provided by the group worked. By which I mean, I was able to extract bacterial DNA, prepare sequencing libraries, produce high-quality sequences, and identify the bacteria they contained. Controls were clean - no detectable amplification. This gives us confidence the DNA we're analyzing came from your tank, not my hands. At every stage these samples and sequencing libraries passed quality control with flying colors; in several ways they were the highest quality amplicon libraries I've ever analyzed. The kinds of bacteria present in these samples were as expected. Most of the tanks share a common bacterial profile.... but there are a few interesting outliers, including a couple tanks that were noted as having problems (e.g. cyano) I'm out of town for a couple days but I'll post results to the website shortly after I get back. Thats how each participant will get their results. I'll also make some posts here discussing specific questions about the data.More soon!

Anyone got a dead or just unused black box you'd like to get rid of? I need some parts for modifying and repairing black box LEDs. Specifically I need the dimmer right now, but decided to just buy old black box units to cannibalize for parts since the right dimmer is apparently not available for sale. The light doesnt have to be functional although of course I'd like to pay less if thats the case Thanks!

Sent a PM - I'd love to get my hands on any fry that die, to test out a new genetic test... I'm not far away, please PM if its something you'd be up for discussing!

Rather than comparing credentials perhaps I can explain this better with data. I often find clarity in exploring data rather than verbal arguments. If the true b=0, there is no difference between your suggestion and the way I'm handling this. (subtracting zero is the same as not subtracting zero) In the real world there is always measurement error, so the intercept is basically never zero. In these cases, the handling of the data matters. For an example, consider a linear relationship with m=6.12, b=-0.025, r2=0.999. Anyone would accept this is a good fit and useful for quantifying the concentration. If you attempt to back calculate the known concentrations from observed absorbance while ignoring the intercept, you'll make bigger errors than if you consider the intercept. You can easily demonstrate this yourself with some toy data. I've pasted some example data below if you'd care to explore this. In this example dataset, ignoring the intercept (your suggestion) leads to a 6% error at the low end and a 2% error at the high end. If you include the intercept you make <3% error at the low end and just under 2% error at the high end (overestimating it in both cases, since the intercept is negative in this example). The effects of ignoring intercepts are always most pronounced at the low end. (Fun debate but lets also keep in mind we are quibbling over 3% vs 6% errors while reading these things with human eyeballs will lead to probably 25% errors ) Example data for fun: C A 2 12 1.5 9.5 1 6.1 0.5 3.05 0.25 1.4 0.125 0.72 0 0 I'll be focusing on barbecuing and stuff like that today but always happy to discuss data analysis later! Have a good 4th, -Eli

You are right of course that in theory the intercept is zero. In theory there is no difference between practice and theory. In practice there is Apologies if this appears argumentative, but in all absorbance or fluorescence based assays it is critical to consider the intercept. It is often not exactly zero for a variety of reasons. Not sure where this misunderstanding arose from. This is the case for literally every chemical assay I've ever worked with or published, e.g. Bradford assay for proteins or fluorometric dye-binding assays for DNA concentrations. Forcing the intercept to zero would be an error that any reviewer would rightly call us out on; you always have to report both slope and intercept. We report and include the intercept because it is often not exactly zero, and forcing it to zero would result in errors by changing the slope. But you are correct that the intercept should be very very close to zero as it is here. So in practice there is little to no difference between our positions, even if there is in theory.

You might want to check out the update - a second dose produced clear evidence of nitrification. The first dose did absolutely nothing but the second worked great. I speculate the first failed due to undetectable residual chlorine from bleach sterilization. Frankly I've always been skeptical of these products on the basis that 1. most (>99%; basically all) marine bacteria cannot be cultured 2. bacteria arent immortal, and the number of viable cells inoculated makes a difference but I cant argue with the round 2 results. It definitely established a nitrifying community. I am curious now to know how that community compares to live rock communities. Extracted DNA today to start finding out...

Its a model 710 from Ebay (i.e. from China). I've just described a bunch of details here

Hi all, I want to share with you how I'm measuring water chemistry for my experiments at AquaBiomics. I've found an affordable instrument that in principle can measure any color-based test kit more precisely and objectively than the human eye. It cost me about $230, so while I won't say its cheap, I think its fair to say its within the budget of the a dedicated hobbyist. The instrument I'm using is the generic "Model 721" widely available on Ebay and AliExpress. https://www.ebay.com/itm/Visible-Spectrophotometer-721-Lab-Equipment-350-1020nm-110V-tungsten-lamp/201962694258 For some reason every picture I've ever taken of mine is absurdly blurry Considering the source I suspect there may be substantial IP violations making this instrument so low priced. But I've worked extensively with several different lab-grade spectrophotometers in my research career. At a basic, functional level, this thing compares to the lab instruments. It has NO bells or whistles. No sipper, no automation, it doesnt even connect to a computer to output data (it has a digital readout, and you write down the number). But it appears to be a fully functional visible wavelength spec. So far I have used this to measure Ammonia, Nitrites, and Nitrates. But it should work for all the usual endpoint color-based tests. (In principle, you could do titrations too, but why bother? The precision of those tests is determined by the accuracy and size of the drops more than visual evaluation of the color) Here I will share details of developing and using those tests. Ammonia I've mostly worked with the Red Sea kit but have recently switched to API. I'll show those API results at the end of this section. The first step for any colorimetric (absorbance based) assay is to identify the wavelength of max absorbance. Make a standard solution of ammonia sulfate, run a test according to the instructions, and run another one on an ammonia-free solution (for the blank; I used NaCl at seawater equivelent concentrations). Then measure its absorbance at a range of wavelengths (blanking the instrument at each wavelength with the blank test). This is a sensible result - there is an obvious peak at 680 nm. I like sanity checking everything, including lab work. The solution is green, and 680 is red, we expect a green solution to absorb in the red. Next, I prepared a stock solution with a known concentration of ammonium sulfate using a precision lab-grade balance. I made a series of dilutions from this to prepare several tubes with known concentrations ranging from 0-2 ppm ammonia. These look like this Blanking the instrument on the 0 ppm solution, I read the absorbance of the others. This relationship is shown here This is a sensible result. The relationship is linear throughout this range of concentrations, and the intercept is nearly zero (i.e. when the concentration is zero, the absorbance is approximately zero). These are good features for a quantitative test. The coefficients for this test are: m=0.59, b=-0.02. So to use the test: I simply follow the instructions with the kit, then put the solution in a cuvette and read it on the spec. I use coefficients from the linear regression to calculate the concentration (sounds harder than it is). i.e. if Absorbance=0.5, I calculate concentration as y = mx + b x = (y-b)/m x = (0.5 - (-0.02)) / 0.59 x = 0.95 (This is all easily handled in a Gsheet spreadsheet with little typing) This has three major advantages over using my eyeballs. 1. It's more objective and consistent. The lighting of the room, how much blue light my eyes have recently been exposed to, the amount of whiskey or coffee I've recently consumed... all of these are likely to influence my brain's subjective evaluation of a color. Not the spec. The spec is a sober and emotionless robot. Given the same solution, it will always give you the same number. 2. Its more precise. The color scales provided with these tests have 6 or so levels (0, 0.25, 0.5, 1, etc.) A human reading the test has to match the color to one of these levels. So you have a precision of ~0.25. Or perhaps at most you allow for "this color is between those two colors", so you can get a precision of half that (~0.13). The spec can tell the difference between 0.102, versus 0.103, versus 0.104, etc. It reports 3 digits and based on repeated readings of the same solution I observe consistent readings to 3 decimal places. So this can detect smaller changes in the substance than reading the test by eye. 3. Its more sensitive. Look at that image. To my old-man eyes, E and F look identical. Honestly, so does D. Maybe a little tiny bit but I'm not sure. The spec is sure, though. Those are the 3 lowest points on the curve shown above. So this can detect lower levels of the substance than reading the test by eye. Complications The ammonia test has an annoying complication. It works great in NaCl solutions but not as great in actual seawater. One of the reagents is a strong base, used to increase the pH for the reaction. In seawater, this leads to precipitation, or cloudiness (you see the same thing when you dose carbonate in your tank). Cloudiness interferes with the spec and leads to weird results. I've been fixing this by putting the samples in the centrifuge for a few minutes before reading them. But this costs time and a disposable centrifuge tube. And few hobbyists have high-speed centrifuges in their fish rooms. I mean, I am a huge geek, but I dont have one at home either. So I recently developed a new tweak to fix this thats quick and easy. Acid. Add 7 drops of 2 M hydrochloric acid after allowing the color to develop. This clears up the cloudiness without any need for centrifugation, and the test remains linear. API test kit This kit is cheaper, so I've recently switched after testing. It has similar properties to the Red Sea kit shown above (although the relationship is different because of the acidification step). I'm acidifying the tests before reading as described above to remove cloudiness. Here is the standard curve for this test; again, nice and linear throughout the range, with a low intercept So thats what I'm using for ammonia from here on. The coefficients for this test are: m=0.29, b=-0.02. Nitrites I used the Red Sea test for this one initially, and have recently started exploring Seachem as an alternative. These data are for the Nitrites test. I'll show the same info as above, without all the text. I used sodium nitrite for the standard curve, prepared with an analytical balance (1 mg precision). The test has an optimum absorbance at 550 nm The tests prepared for a standard curve from 0-2.5 ppm look like this. Check out those invisible colors at the low end. Again the spec has no problem detecting them. Here is the standard curve -- the coefficients for this one are: m=0.25, b=-0.01. My only complaint about this test is that it maxes out the absorbance pretty low (~6 ppm) so different dilutions are required to measure higher values, which is a pain. Of course, its rare NO2 reaches anything higher than "undetectable" in a mature tank so a minor issue. Nitrates Finally NO3. The only complication here is that as far as I understand it, this test actually measures the sum of NO2 + NO3 -- it converts existing NO3 into NO2 then measures the sum of NO2 + NO3. So I run this test as instructed, using a standard curve as above, then subrtract the NO2 data The absorbance spectrum matches NO2, as you would expect (since its actually measuring NO2). Peak at 550 Tests prepared on a series of known concentrations (0-8 ppm) of sodium nitrate looked just like the NO2 tests above, except paler. The standard curve looks like this, with coefficients: m= 0.03, b = 0.00. ---- So thats how I'm measuring water chemistry in the lab. I'll be adding other tests as time goes on. I'm especially curious to see how sensitive phosphate tests are using this instrument, since this is one of the few materials present at low levels where we really care about the concentration.

I have an update that totally changes my conclusions about the bacterial product. I couldn't believe the total lack of an effect the first time. So I added an additional dose of bottled bacteria to both of the B tanks, and sampled the microbiome again in each of the B tanks and the D tanks (dry rock controls). Then I measured water chemistry as usual. I only have ~4 days of data in hand but its clear that it worked this time, very convincingly. Here are the ammonia consumption data for the new trials, plotted along with the previous live rock data for comparison. I calculate nearly identical rates of ammonia consumption for bottled bacteria vs live rock (~45% per day over the first 4 days of data). Like before, all data shown here represent the average from duplicate identical tanks in each group. And there is convincing evidence this consumption results from nitrification. Here are the data for nitrite and nitrate accumulation. The bottled bacteria produce even more than the live rock tanks (again, I've plotted the new B and D trials alongside the older live rock data) Its always frustrating when you try to repeat an experiment and get different results. In my research life we used to say "thats why you never repeat an experiment!" [This is a scientist joke; reproducibility is obviously important but theres this annoying tendency for things to go differently the second time... ] But I am relieved. So many people are using these products, they couldnt be totally snake oil. They obviously worked great in my second trial. So this is good, I can focus on the original question for those treatments instead of having to work hard to prove the product didnt work So why did the first inoculation fail while second inoculations from the same bottle worked? I think there may have been residual chlorine in the tanks from bleach sterilization. I neutralized with thiosulfate and tested to confirm the absence of chlorine, but using a fairly insensitive kit. My sensitive kit arrived 2 days into the experiment and also showed zero. So I thought all was OK, but I cant rule out the presence of tiny amounts of chlorine for the first 2 days... it would make sense that this would affect suspended bacteria more than biofilms, and the product instructions do emphasize the sensitivity to chlorine. In conclusion - I now have three groups of functional nitrifying communities to analyze, in addition to the control. Next steps are to finally analyze these samples I've been accumulating! -Eli

I'm skeptical enough about the negative result on the bacterial product that I went ahead and dosed those tanks again with bacteria and ammonia. I'll run another time course with this fresh dose. I will absolutely be including both these tanks and the bottle itself in my first batch of samples (preparing this batch now). I can say for sure the bottle and cold pack were cold on arrival and I've kept them cold since arrival... but cant vouch for its history beyond that. If I get another round of negatives I will buy another bottle to triple check.

Yeah , it surprised me too. I think its because I took such pains to avoid contamination. The tanks are covered, and I only work on them with sterilized gloves. Hard to argue with 4 tanks that completely lack nitrification after a month, though! I was equally surprised that bacteria in a bottle did absolutely nothing. My expectation was that these would promote nitrification but do little for bacterial diversity. SO far all the evidence in hand points to a placebo effect... And thanks for your comments on NO2 toxicity or the lack thereof, that matches my understanding. Seems to me thats an argument in support of considering NO3 appearance as the endpoint rather than NO2=0.

Thanks, you bring up another endpoint I should measure - returning to zero nitrites. One of my 4 live rock tanks has returned to zero... I'll go ahead and quantify that difference too. Perhaps that endpoint will show a difference between LR sources. (The timing of NO2 and NO3 appearance was pretty much identical in both, but I'm skeptical they're really gonna behave the same).

Fixed now as far as I can tell! These icon-driven interfaces always mess me up.

Hi everyone. I've just completed a month-long experiment in my new lab at AquaBiomics at thought you might be interested in reading about it. The subject is a familiar one: cycling a tank. In other words, establishing a nitrifying community of bacteria (capable of converting toxic ammonia into nitrites, and then into the relatively non-toxic nitrates). Hobbyists have been doing this for decades, and in recent years have been measuring seawater chemistry to indirectly measure the development of these communities. I've repeated this, following more or less standard practices, in order to directly measure the changes in microbial communities at each stage. Along the way, I've dosed with ammonia and measured water chemistry to monitor development of the tanks. These are the results I have to share. My study includes several features not often included in the "forum literature": 1. replication - I prepared all experimental tanks at the same time using identical materials, and set up duplicate identical tanks in each group 2. controls - normal people don't invest time and resources setting up tanks in a way they know won't work. I did this here, because controls are important if we want to draw confident conclusions about the effects of our practices. 3. semi-sterile environment - I set up my experimental tanks in a way that minimizes contamination from environmental bacteria, and took great pains to avoid any contamination of the tanks during sampling and maintenance. What I did Built experimental tanks For this and other studies I set up an array of 12 experimental tanks. For these experiments I built custom all-in-one nano tanks (20 gal), designed to minimize cross contamination between systems. They're not designed to be pretty display tanks, but a couple months in I'm happy with their performance. Here are shots of one tank during testing, with a top view during construction to show how it works. Its an all-in-one design, with a two-chambered sump in the back. Water flows from display, then down through live rock chamber, then up into the return pump/heater chamber, then back into the display. The design reflects an effort to minimize costs so I could afford 12 independent systems, minimize contamination from the environment into the tank, and to fit the essential reef keeping functions in a small space. Here are the tanks on shelves (dry in the pictures but they've actually been wet almost 2 months). I fit 12 of these mini reefs into my little lab in Junction City. Started tanks four different ways To measure the effects of different practices for cycling a tank (here, I'll use this to mean "establishing a community of bacteria capable of metabolizing ammonia into nitrate"), I set up duplicate tanks in each of four different ways: F - maricultured rock from Fiji (artificial base rock) T - maricultured branch rock, Tonga (real coral skeletons) D - dry Pukani base rock (acid washed and neutralized then soaked in hydrogen peroxide) B - dry rock (as D) with a popular "bacteria in a bottle" product I first sterilized each tank and all pumps. etc with alcohol and bleach, then filled each tank with sterile-filtered (0.2 µm) UV-sterilized artificial seawater. I added 4 lbs sand to each tank, which I sterilized using hydrogen peroxide prior to adding to the tank. I used the bacterial inoculant product (treatment B) according to the manufacturers instructions. I'm not gonna name the product here because my results are not terribly flattering, but it is advertised as establishing and maintaining a biofilter capable of converting ammonia into nitrate. Here are images of the rock I used. I went to great pains to locate something as close as possible to real live rock (which is sadly very hard to come by). The Fiji rock had absolutely beautiful growth. The rock underneath is kind of garbage; its a mixture of cement, sand, and a little shells and rubble... with that ugly artificial purple color added. I'd rate this rock A for growth, F for the rock itself. Very pretty and very real in terms of the biology, but very fake in terms of the rock itself. All the visible growth died shortly after arrival (even though I maintained the rock at temperature with circulation and skimmer, with regular water changes to minimize dieoff). But presumably the bacteria remain. The Tonga rock had a mixture of coralline ranging from pale to bright purple. No other visible life, but I consider live coraline a good indicator of live bacteria. Its very, very porous stuff, real branch rock from old dead coral branches. Its also pretty small and brittle, would be challenging to aquascape with in a big tank. But great for my purposes. Really, I'd consider this ideal sump rock material: porous, and relatively fresh from the ocean. The dry Pukani was also hard to come by. I ordered all in stock that I could find online, then a little more from local stores. I sterilized this stuff pretty rigorously with first and acid wash (neutralized with baking soda), then soaking for 2 weeks in hydrogen peroxide, with new H2O2 added several times. I doubt it was surgical-level sterile when I added it to the tanks, but it was as close as I could make it without an autoclave. Added ammonia to fuel bacterial growth I dosed ammonia to 1 ppm in each tank using ammonia sulfate. I measured the initial levels after dosing, then measured ammonia levels weekly and dosed additional ammonia to maintain 1 ppm in each tank throughout the experiment. Monitored changes in water chemistry I measured ammonia, nitrite, and nitrate levels throughout the experiment using hobbyist kits, and read the results using a spectrophotometer. This is a subject I'll write up in a separate post, because it deserves the space. For now, I'll keep it brief. The spec works as an electronic eye, and reads the kits in the same way as a human. Except it can tell the difference between fine shades of green or pink that are invisible to the human eye. I ran these tests according to the kit instructions, then transferred the solution into a cuvette for reading in a spectrophotometer. I used Red Sea kits for all three. [After using these for a month or so, I can say I trust the data from these kits but am exploring other tests currently. The ammonia test has more cloudiness than I would like, and there are cheaper alternatives for NO2/NO3. ] In a subsequent post I'll describe the details of these measurements for those who are into that sort of thing. Collected samples for microbiome analysis I collected samples from each tank weekly throughout the experiment, using the same process and supplies included in the AquaBiomics sampling kit. I haven't analyzed these yet; that'll be part 2. What I found Falling Ammonia levels are an unreliable indicator of nitrification This is a minor point, but maybe worth noting. During the first few weeks of the experiment, ammonia was consumed in all tanks. Once a week I measured ammonia levels in all tanks and dosed additional ammonia to maintain levels at 1 ppm. Based on these data, we can estimate ammonia consumption rates for each tank in each week, revealing that a large fraction of the ammonia was consumed in each tank (43-92%). For example, the data for week 2 are shown in Figure 1. Despite the consumption of ammonia during the early weeks, there was no evidence of nitrite or nitrate production in most tanks (Figure 2-3). Although initially confusing, this actually makes sense if we remember that ammonia is a valuable nutrient and is rapidly taken up by lots of organisms in the ocean. Its not all oxidized into nitrite and nitrate by bacteria. Some of the ammonia went into algal biomass, because during the first week I foolishly left the lights on a 12:12 cycle. This promoted a little bit of microalgal growth in the LR tanks, and algae consume a lot of ammonia. And some of it went into bacterial biomass - I observed visible bacterial film growth in most tanks. In the final week of the experiment, I measured NH4 every 2 days after dosing the tanks at ~1 ppm. Figure 4 illustrates how rapidly ammonia is consumed in tanks with nitrifying bacterial communites (F, T) and how slowly it's consumed in dry rock tanks (D, B). It may also be useful to view the complete time series for NO2 and NO3 (Fig 5 and 6). Live rock rapidly establishes a nitrifying community My results demonstrate what we already knew -- but I like having numbers and controls supporting this conclusion. The tanks started with live rock clearly established nitrifying communities within the first 2 weeks, based on the production of NO2 and NO3 along with consumption of NH4 (Figs 2, 3). I'm not adding livestock to anything yet, but these tanks would be ready to stock within 2 weeks of starting the tank (and likely within the first week). I see no convincing evidence of a difference between these live rock sources at the level of nitrification. This surprises me considering the difference in quality of the base rock. I anticipate their microbiomes will be very different based on their sources; it will be interesting to see what they have in common. Without environmental contamination, dry rock cycles very slowly This one was a little more surprising. Many of us have started tanks with live rock and successfully cycled tanks started with dry rock. I've always seeded it with a little live rock, but I know many have started them without any live rock source. And eventually (weeks to months) these tanks are expected to cycle. In contrast, I find no evidence of nitrification in my dry rock tanks. With the lights off, ammonia levels remain nearly constant in these tanks, and after a month of maintaining 1 ppm ammonia there is no evidence of NO2 or NO3 in the tanks (Fig 5, 6). How can we explain this contrast? I believe the explanation is that I took great care to avoid environmental contamination in these tanks. I sterilized the tanks, the water, the sand, and the rocks prior to starting each tank. The tanks are covered to minimize dust and aerosols. And I only handle the tanks with bleach-sterilized gloves or tubes. No sensible hobbyist starts a tank like that! But it provides an important control for my experiments, since it demonstrates that bacterial communities in my other treatments were produced by the treatments themselves rather than environmental contamination. The implication for the hobby is - its important to get a new tank dirty. You really can make them too clean - I've just done it. Some bottled bacterial products provide no measurable benefit This one will seem obvious to some and complete nonsense to others. But the data in hand are pretty convincing for me. Although I used this product as directed by the manufacturer, and set these up side by side with dry-rock controls, the tanks started with a bacterial product also show no evidence of nitrification at all, after 1 month (Fig 4, 5, 6). Just like the dry rock tanks, the bacteria in a bottle tanks did not deplete ammonia in my time series, and show no evidence of NO2 or NO3 after a month with NH4 at 1 ppm. I don't want to over-generalize. But I have to conclude that in this case, the product appeared to have no measurable benefit for establishing a nitrifying community. Why did it have no benefit for me, while many have successfully started tanks with these products? I would gently suggest that many peoples' experiences with these products were based in non-sterile environments, and lacked replication or controls (otherwise identical tanks started without the product). I suspect that the bacterial communities established in standard hobbyist environments (like my own tanks in my living room) are seeded from environmental sources, whether bottled bacteria are added or not... It will be interesting to see what the microbial analysis of these tanks over the past month reveals! And I'm starting additional experiments now to follow up on these results. I'll be curious to hear how these results compare with your own experiences.

Hi all, Anyone here currently breeding clownfish? If so I would like to discuss 1. asking for samples of culled/dead fry for a new genetic test I've developed 2. buying 12 young clownfish (my goal is to minimize price here, since I need 12. neither breed nor sex matters, just that they're from the same clutch). Thanks for looking, -Eli

I'm selling a standard 55g tank purchased from Petco about 4 years ago. No problems, just upgraded. Looking for $50.

Sorry for the slow updates, been very busy. I believe I have all samples in hand for people from this forum. Still waiting on a few others. My best update is - I am nearly done with the big cycling experiment, and have a bunch of samples in hand from that! The good news thats obvious right away is that the tanks behaved very differently in terms of nutrient cycling, so it appears I was able to avoid major problems with cross contamination. I'll post real updates in the next few days...

This stuff is like magic. I had a batch of zoa frags I'd neglected that were totally covered in blue cloves -- couldnt even see the zoas anymore. I dipped them in a strong dose of this product for about an hour, then rinsed and returned to the tank... I checked them today and theyre clean again, all the BCP died and the zoas are happy. I've been too nervous to treat a whole tank because I have lots of weird inverts I dont want to kill. But the dip worked so well, I think I'm gonna start dipping rocks. (I am one of those foolish people who actually bought BCP once upon a time... )

I have an educational account with a wholesaler, who imports this from a mariculture facility in Tonga. I have also been frustrated by the lack of actual live rock in the market currently! On that note, I recently also bought some of this stuff https://www.liverocknreef.com/catalog/live-rock-specials-with-shipping-included./fiji-saltwater-live-rock-7915.html Its by far the nicest live rock I've seen in the last five years, it has all the growth of real live rock, thick sponges and coralline and tunicates. The only downside is that its fake rock (cement / rubble / shells) instead of actual dead corals. If anyone else is looking for a bunch of live rock with actual life on it, that stuff is great. What I am selling here is genuine live rock (dead corals with encrusting growth on it), but not as much growth as the Fiji stuff. Still, beautiful colors and lots of life. I have several potential buyers lined up in PMs (first come first serve), so I am pretty sure this is sold.