Effects of tap water on Nitrifying during Rip-Clean method: Experiment

[ApoIsland]

Gotcha I guess that could be it. Was hoping for maybe a paper or article showing something more complex explaining how it could live. I have scrubbed areas of coraline where it looked like color came off a bit but bounced back pretty quickly. And the areas where I've scrubbed and quick rinsed algae away coraline under or around scrubbed areas covered with coraline pretty quickly.

An easy test for you would be to pull one of those beautifully purple covered rocks you have and then dip half the rock in peroxide. See if you can tell the difference between dead and live coraline after a few days.

 

[taricha]

Most things will be similar if not the same - except:

1. Will use 2 liquid tests (One an api)
2. Will add a saltwater solution with 2 PPM ammonia at the start of each test.
3. Will use more gentle types of 'rinse and scrub' first and extend onto more 'aggressive'

As I said will be out of town this weekend - so it will start next week - so anyone with comments feel free to make them.

PS - so - even though the rocks have been in the tank for multiple years - my strong guess is that they are not going to be able to process all a full amount (2 ppm) ammonia on day 1. That it will need to build up.

here are a couple questions though - up front:

1. Use rock from the tank (which Is lit about 12 hours/day) or Use the rock from the sump - which is basically always in the 'dark'
2. Here is another Step 1, 2, and 3

Step 0. Baseline measurements from the tank (No2, No3, ammonia, PO4, temp, Specific Gravity, pH)
Step 1a. Day 0 - add New Saltwater, Rock (1 lb/gallon) powerhead for flow, Same temp and lighting as the tank. Day 1- add .25 ppm ammonia Day 2 - add .5 ppm ammonia Day 3 add 1 ppm Ammonia Day 4 add 1 ppm ammonia. Determine how quickly the ammonia rises (or is processed). Perform the measurements - same as step 0 - each day. Wait until all ammonia is processed to 0. Hopefully by day 5.
Step 2 (Same as old step 1) Add 2 ppm ammonia with fresh water and keep repeating this until 2 ppm is processed in about 24 hours.

Step 3 (Same as old Step 2) - same as before - same measurements

Some questions - should phosphate be added if at a certain level?
Should pH be adjusted?

I'll address quick Qs first.
@Dan_P looked for low PO4 effect in halting nitrification. My over-the shoulder interpretation of his data is that It was present but really insignificant. As long as PO4 is not zero, should be okay. 0.10ppm PO4 fine.

pH will be fine on its own. It'll be stabilized by the aragonite buffer of the rock in the test. Nitrification works just fine at 8.4 or ~7.6 (at the aragonite buffer). Biospira still nitrified for me just fine at pH of closer to 7.0

Why do this as a lighted test? That conflates nitrification and photosynthesis. If you want to test literal nitrification, leave it in the dark (regardless of where the rock to be tested came from). If you want to test just general ammonia uptake, then lighting the same as the tank is fine - but you won't be looking at "nitrification" in the strict sense. And the photosynthetic processes may dominate.

Getting further into the weeds, if you are trying to distinguish between dead nitrifiers and merely salinity-shocked nitrifiers, I don't think you can with porous reef rock. Because whether the biofilm is just salinity shocked, and then recovers or is dead and reseeded by deeper bacteria that weren't killed - it'll look the same: Short term total halt to nitrification and a recovery to whatever the target level is over some days.


(If these still aren't the things you were looking for feedback on then point me in the right direction)

 

[sixty_reefer]

I'll address quick Qs first.
@Dan_P looked for low PO4 effect in halting nitrification. My over-the shoulder interpretation of his data is that It was present but really insignificant. As long as PO4 is not zero, should be okay. 0.10ppm PO4 fine.

pH will be fine on its own. It'll be stabilized by the aragonite buffer of the rock in the test. Nitrification works just fine at 8.4 or ~7.6 (at the aragonite buffer). Biospira still nitrified for me just fine at pH of closer to 7.0

Why do this as a lighted test? That conflates nitrification and photosynthesis. If you want to test literal nitrification, leave it in the dark (regardless of where the rock to be tested came from). If you want to test just general ammonia uptake, then lighting the same as the tank is fine - but you won't be looking at "nitrification" in the strict sense. And the photosynthetic processes may dominate.

Getting further into the weeds, if you are trying to distinguish between dead nitrifiers and merely salinity-shocked nitrifiers, I don't think you can with porous reef rock. Because whether the biofilm is just salinity shocked, and then recovers or is dead and reseeded by deeper bacteria that weren't killed - it'll look the same: Short term total halt to nitrification and a recovery to whatever the target level is over some days.


(If these still aren't the things you were looking for feedback on then point me in the right direction)

Are you aware if there is any po4 in a fresh batch of salt mix?

 

[taricha]

Do you know if there is any po4 in a fresh batch of salt mix water?

In fresh Instant Ocean it's low but not zero (on hanna ULR P) and the nitrification of biospira proceeds just fine without any P addition.
Additionally @Dan_P fed biofilms some ammonia water that had 0.0 or 0.2ppm PO4 and barely detected a difference in the ammonia processing rate. (Maybe Dan will slap me around if I'm mischaracterizing that summary.)

The idea of P starvation halting nitrifiers turns out to be hard to reproduce.

 

[DrZoidburg]

Are you aware if there is any po4 in a fresh batch of salt mix?

Name of product tested: Red Sea Salt Batch number:**********

Tested element Units Acceptable Range Batch Analysis pH 8.2-8.5 8.28 Total Alkalinity meq/l 2.7 - 3 2.95 Calcium | Ca mg/l 415-445 428 Magnesium | Mg mg/l 1240-1320 1270 Sulfur | S mg/l 870-930 890 Potassium | K mg/l 375-405 376 Bromine | Br mg/l 60-70 65.04 Strontium | Sr mg/l 8-9 8.42 Boron | B mg/l 4-4.4 4.27 Total ammonium | NH4/NH3 mg/l ≤0.1 ≤0.1 Nitrate | NO3 mg/l ≤0.3 ≤0.3 Phosphate| PO4 mg/l ≤0.03 ≤0.03

 

[DrZoidburg]

Everyone involved looks good. I wonder if this is a scrub, rinse, or city tap water issue. I said way way before I used certain bacteria to cycle rocks. They were started at 0.00 well water, and never exposed to anything crazy except gradual salinity increase. Not the other way around like this thread. This is how I started almost every cycle I've done. Response to a question earlier. Any coralline I've hit with anything aggressive always turns white, then eventually come back. I always assumed it died, then gets used as a substrate for new spores. Agree no definition to "rip clean" as some one said earlier it is not a new idea. Been doing this for a while also but very differently.

 

[Coxey81]

Everyone involved looks good. I wonder if this is a scrub, rinse, or city tap water issue. I said way way before I used certain bacteria to cycle rocks. They were started at 0.00 well water, and never exposed to anything crazy except gradual salinity increase. Not the other way around like this thread. This is how I started almost every cycle I've done. Response to a question earlier. Any coralline I've hit with anything aggressive always turns white, then eventually come back. I always assumed it died, then gets used as a substrate for new spores. Agree no definition to "rip clean" as some one said earlier it is not a new idea. Been doing this for a while also but very differently.

You reminded me, someone wanted to see what happened to the coralline... was it you @Garf?

Before first, after second

I'm not sure it changed a ton. Tilting the camera can take it from a pretty bright green to a faded one.

 

[DrZoidburg]

@Coxey81 That green stuff maybe just a type of boring algae though. Literally every dry rock start I've done gets that then it slowly goes away. Can see a difference though

 

[Coxey81]

@Coxey81 That green stuff maybe just a type of boring algae though. Literally every dry rock start I've done gets that then it slowly goes away. Can see a difference though

Pretty sure it's green coralline. It doesn't scratch off and the guy I got the tank from said he dosed it and that's what it was.

Purple is over taking it in my tank though.

 

[DrZoidburg]

The green stuff plates out kind of though, and likes lower salinity. Where as boring types thrive in newer tanks, or older tanks with imbalances. It could be other things too just a guess I guess.

 

[Coxey81]

The green stuff plates out kind of though, and likes lower salinity. Where as boring types thrive in newer tanks, or older tanks with imbalances. It could be other things too just a guess I guess.

Eh, I don't really care either way. It's getting covered up anyway, lol.

 

[Dan_P]

In fresh Instant Ocean it's low but not zero (on hanna ULR P) and the nitrification of biospira proceeds just fine without any P addition.
Additionally @Dan_P fed biofilms some ammonia water that had 0.0 or 0.2ppm PO4 and barely detected a difference in the ammonia processing rate. (Maybe Dan will slap me around if I'm mischaracterizing that summary.)

The idea of P starvation halting nitrifiers turns out to be hard to reproduce.

Yes, the PO4 effect was small. I had assumed it was really important. It might be a case that there was enough PO4 in Instant Ocean to support nitrifying bacteria growth. I am repeating these experiments with free flowing aragonite sand and small aragonite slices. I should take a look at the tiny PO4 effect again.

 

[Lasse]

day 1

08-11-21
Rock submerged
Tank cleaned
85% water change
24:00 local time 2.2 ppm ammonia added

day 2
09-11-21
24:00 local time tank tested 0.2ppm ammonia

24:05 local time 6 drops added
24:20 tested 2.4 ppm

10-11-21

17:00 local time 0.1ppm
18:00 local time 0 ppm
19:00 local time 10 drops added
19:30 tested result approx 3.6


11-11-21

19:00 local time 0ppm

total ammonia over 66 hours

22 drops or 8ppm approximately

hope it helps tried to be the most precise as possible

OK - you have added total 8 ppm NH4 -> 14/18 * 8 = 6.22 ppm NH4-N (ammonium nitrogen) if all should be converted to nitrate nitrogen NO3-N it should be 6,22 ppm (NO3-N) or as NO3 -> 62/14 * 6,22 = around 27 ppm NO3.

When your NO2 is close to 0 - measure your nitrate concentration and calculate how much less NO3 you have. Let us say that your measurements show 5 ppm Nitrate -> 5*14/62 = 1.13 ppm NO3 - N. If all your NH4-N should been converted into NO3-N it should have been 6.22 ppm NO3 -N. You got 1.13 ppm NO3-N - it means that nearly 5 ppm NH4-N has not been converted into nitrate instead probably have been taken up by photosynthetic organisms - as NH4 it will be 18/14*5 ppm NH4 -> 6.4 ppm NH4 that have been taken up by organisms different from the nitrification - probably by your aiptasia and micro algae

Sincerely Lasse

 

[MnFish1]

I'll address quick Qs first.
@Dan_P looked for low PO4 effect in halting nitrification. My over-the shoulder interpretation of his data is that It was present but really insignificant. As long as PO4 is not zero, should be okay. 0.10ppm PO4 fine.

pH will be fine on its own. It'll be stabilized by the aragonite buffer of the rock in the test. Nitrification works just fine at 8.4 or ~7.6 (at the aragonite buffer). Biospira still nitrified for me just fine at pH of closer to 7.0

Why do this as a lighted test? That conflates nitrification and photosynthesis. If you want to test literal nitrification, leave it in the dark (regardless of where the rock to be tested came from). If you want to test just general ammonia uptake, then lighting the same as the tank is fine - but you won't be looking at "nitrification" in the strict sense. And the photosynthetic processes may dominate.

Getting further into the weeds, if you are trying to distinguish between dead nitrifiers and merely salinity-shocked nitrifiers, I don't think you can with porous reef rock. Because whether the biofilm is just salinity shocked, and then recovers or is dead and reseeded by deeper bacteria that weren't killed - it'll look the same: Short term total halt to nitrification and a recovery to whatever the target level is over some days.


(If these still aren't the things you were looking for feedback on then point me in the right direction)

These were the comments I was looking for

 

[Coxey81]

Taken 48 hours after rinse and .75ml(3ppm) dose

NH4+NH3 = ~1.5 ppm

NO2 = zero ppm

 

[sixty_reefer]

Taken 48 hours after rinse and .75ml(3ppm) dose

NH4+NH3 = ~1.5 ppm

NO2 = zero ppm

At this rate is looking at 4 days do deplete, I believe your first run depleted in 3 days wasn’t it? But you had a larger amount of ammonia I believe it was 1.1ml (16 drops) and you used 0.75ml on this run. it’s a bit earlier for maths but isn’t this looking to go towards the same results as your first 3 day run?

 

[sixty_reefer]

@Lasse thank you for the calculation is good to know, as you pointed out earlier my experience doesn’t fully mimic @Coxey81 experience, I’ve made that decision because I was interested in a more real life test under extreme conditions. I always had concerns about cleaning a full tank for things that can be resolved without having to do so, I am from a believer that you don’t have to break down a tank to eradicate dinoflagellates, pest algae’s, Aipatasia etc… we’ve been dealing with them inside our tank for years. There for my choice, I never intend to evaluate one aspect of a largerl picture. I now believe I got enough information to explain why Breakin down a tank is not always successful, there is things that need looking at to avoid dismantling a full tank just to encounter another problem once re assembled. Will give my final update in 11.30 hours.

 

[Lasse]

@Lasse thank you for the calculation is good to know, as you pointed out earlier my experience doesn’t fully mimic @Coxey81 experience, I’ve made that decision because I was interested in a more real life test under extreme conditions. I always had concerns about cleaning a full tank for things that can be resolved without having to do so, I am from a believer that you don’t have to break down a tank to eradicate dinoflagellates, pest algae’s, Aipatasia etc… we’ve been dealing with them inside our tank for years. There for my choice, I never intend to evaluate one aspect of a largerl picture. I now believe I got enough information to explain why Breakin down a tank is not always successful, there is things that need looking at to avoid dismantling a full tank just to encounter another problem once re assembled. Will give my final update in 11.30 hours.

You have to take my calculations between the thumb and index finger- but it shows how you do a mass calculation. What´s unchanged in this chain is the amount of N atoms - they do not disappear. Hence the conversion to N atoms (or their equivalent). The atomic weight of NH4 is 18 (N=14; H4 = 4*1) - the amount of N in NH4 is therefore 14/18 * X g, atomic weight of NO3 is 62 (N=14; O3 = 3*16) -> The amount of N in NO3 is 14/62 * X g

If you have 1 ppm of NH4 in water (ppm ≈ mg/L) the mg/L of N is 14/18*1 ≈ 0.78. You express this as NH4-N. Now - if all of this N goes further to NO3 it is still 0.78 N and in order to get NO3 we have to multiply 0.77 with 62/14 which give us 3.41 mg/L of NO3

Sincerely Lasse

 

[Coxey81]

At this rate is looking at 4 days do deplete, I believe your first run depleted in 3 days wasn’t it? But you had a larger amount of ammonia I believe it was 1.1ml (16 drops) and you used 0.75ml on this run. it’s a bit earlier for maths but isn’t this looking to go towards the same results as your first 3 day run?

First run was 16 drops which should be close to 4ppm based on 1ml for 16 drops for me.

It took 99.5 hours, or 4 days and 3.5 hours.

This run is .75ml (3ppm)

So, right now, not looking as good. But not a whole lot worse. I think this will take over 3 days, but not 4. I'll try to get it down to the hour.

 

[sixty_reefer]

You have to take my calculations between the thumb and index finger- but it shows how you do a mass calculation. What´s unchanged in this chain is the amount of N atoms - they do not disappear. Hence the conversion to N atoms (or their equivalent). The atomic weight of NH4 is 18 (N=14; H4 = 4*1) - the amount of N in NH4 is therefore 14/18 * X g, atomic weight of NO3 is 62 (N=14; O3 = 3*16) -> The amount of N in NO3 is 14/62 * X g

If you have 1 ppm of NH4 in water (ppm ≈ mg/L) the mg/L of N is 14/18*1 ≈ 0.78. You express this as NH4-N. Now - if all of this N goes further to NO3 it is still 0.78 N and in order to get NO3 we have to multiply 0.77 with 62/14 which give us 3.41 mg/L of NO3

Sincerely Lasse

First run was 16 drops which should be close to 4ppm based on 1ml for 16 drops for me.

It took 99.5 hours, or 4 days and 3.5 hours.

This run is .75ml (3ppm)

So, right now, not looking as good. But not a whole lot worse. I think this will take over 3 days, but not 4. I'll try to get it down to the hour.

According to my math 3 days and 2:55 or 74.55 hours total to match your first run. Not sure what that it means but your rinse looks close from the first test so far in ammonia depleted. Has no2 and no3 been giving you similar results from first test?
It does look like your rocks went back to original capacity from wend they were removed from the sump.