New Nitrifying Bacteria Experiment.

[MnFish1]

SO - TODAY'S UPDATE - if anyone cares lol

A bit of user error. (well technology error) - ATO Mess up - I have no saltwater to make a new experiment (Failed float switch)

IE - NO start of experiment today.

 

[MnFish1]

Because of technology issues - I've had a couple days to think on the experiments, etc thus far, and I am going to change focus a bit - Because after looking at all the data - it is clear that (in this set up):

1. Rock taken from a tank (whether from a dark sump - or from a lit area of a display tank) - still has residual capacity to process ammonia over a week or so. I.e. the rock is not saturated with bacteria or ammonia consuming organisms.
2. Rock from a tank containing rock from a lit tank processes ammonia more quickly than rock from a dark sump.
3. The Rock from a lit tank does NOT process ammonia more quickly due to photosynthesis. EDIT - Though there may be differences in surface area on the rock in the formerly lit tank as well.
4. Multiple salt-water rinses with fresh water do NOT affect the ability of the rock to process ammonia
5. The glass, HOB filter, powerhead do not seem to be responsible for the increase in ammonia processing capability.
6. The 'Density'/'Porosity' of the rock MAY play a role in the differences seen between tanks in item 2 above - BUT - over time, the sump rock and lit rock both can process the same amount of ammonia over the same time period.
7. The current 'rock/water' is adequate to process 2 ppm ammonia in 24 hours repeatedly. Thus, meeting the criteria for Dr. Tim's 'cycle'. And thus - also in all likelihood the ratio of rock to water is similar to amounts in many reef tanks (about 1.5 lbs/gallon). Thus - at least in part - simulating a 'rip-clean'.
8. What amazes me is that despite multiple complete tank water changes - the water is crystal clear. No blooms, etc - and I mean - it is totally crystal clear. This was even when the light was on for multiple days in tank 2.
9. The Seachem alert badge continues to work 'appropriately'. The API tests - have not shown any 'false 0.25 ppm ammonia readings. In each case (1) - where I thought this was the case - 12 hours later it read 0.

So - now for the changes:

Todays experiment (#7) Protocol:

1. Empty tanks, wipe sides.
2. Scrub rock in tank water - for 5 minutes. Rinse rock with clean salt water Once
3. Add Rock and Clean water back to tank - do baseline measurements (because its a new batch of Saltwater)
4. Add 2 ppm ammonia to water - and do baseline measurements.
5. Measure ammonia as before

If 2 ppm is processed in 24 hours - will repeat the experiment again - to see if doing it more than once will affect processing capability (and also act as a replicate.

Thereafter - will skip RODI rinsing - and go directly to Tap water.

In the meantime - if anyone has any comments - suggestions - requests - feel free. I have not been able to obtain a low nitrite reader - but working on it @Lasse.

@Lasse, @Coxey81, @sixty_reefer, @Dan_P, @Garf, @brandon429, @Rmckoy, ETC ETC

 

[Garf]

Because of technology issues - I've had a couple days to think on the experiments, etc thus far, and I am going to change focus a bit - Because after looking at all the data - it is clear that (in this set up):

1. Rock taken from a tank (whether from a dark sump - or from a lit area of a display tank) - still has residual capacity to process ammonia over a week or so. I.e. the rock is not saturated with bacteria or ammonia consuming organisms.
2. Rock from a tank containing rock from a lit tank processes ammonia more quickly than rock from a dark sump.
3. The Rock from a lit tank does NOT process ammonia more quickly due to photosynthesis.
4. Multiple salt-water rinses with fresh water do NOT affect the ability of the rock to process ammonia
5. The glass, HOB filter, powerhead do not seem to be responsible for the increase in ammonia processing capability.
6. The 'Density'/'Porosity' of the rock MAY play a role in the differences seen between tanks in item 2 above - BUT - over time, the sump rock and lit rock both can process the same amount of ammonia over the same time period.
7. The current 'rock/water' is adequate to process 2 ppm ammonia in 24 hours repeatedly. Thus, meeting the criteria for Dr. Tim's 'cycle'. And thus - also in all likelihood the ratio of rock to water is similar to amounts in many reef tanks (about 1.5 lbs/gallon). Thus - at least in part - simulating a 'rip-clean'.
8. What amazes me is that despite multiple complete tank water changes - the water is crystal clear. No blooms, etc - and I mean - it is totally crystal clear. This was even when the light was on for multiple days in tank 2.
9. The Seachem alert badge continues to work 'appropriately'. The API tests - have not shown any 'false 0.25 ppm ammonia readings. In each case (1) - where I thought this was the case - 12 hours later it read 0.

So - now for the changes:

Todays experiment (#7) Protocol:

1. Empty tanks, wipe sides.
2. Scrub rock in tank water - for 5 minutes. Rinse rock with clean salt water Once
3. Add Rock and Clean water back to tank - do baseline measurements (because its a new batch of Saltwater)
4. Add 2 ppm ammonia to water - and do baseline measurements.
5. Measure ammonia as before

If 2 ppm is processed in 24 hours - will repeat the experiment again - to see if doing it more than once will affect processing capability (and also act as a replicate.

Thereafter - will skip RODI rinsing - and go directly to Tap water.

In the meantime - if anyone has any comments - suggestions - requests - feel free. I have not been able to obtain a low nitrite reader - but working on it @Lasse.

@Lasse, @Coxey81, @sixty_reefer, @Dan_P, @Garf, @brandon429, @Rmckoy, ETC ETC

Excellent stuff. So by previous tests, existing literature, this scrubbing of everything should have the greatest effect (apart from scalding hot water), unless your test throws us a curve ball. Can’t wait

 

[MnFish1]

EXPERIMENT 7 BASELINE

1. Tank 1 Sump rock, Dark, Rinsed scrubbed x about 2-3 minutes/rock (entire surface)
2. Tank 2 Tank Rock, Dark, Rinsed, scrubbed x 2-3 minutes on entire surface.
3. Both tanks drained, detritus removed
4. Rocks Rinsed with FSW then New FSW added to each tank - Baseline measures taken (Below)
5. 2 ppm ammonia added - Post ammonia pictures below.

Sump Rock




Tank Rock


Tanks Post rinsing

Tanks after ammonia

Ammonia Pre-addition

Ammonia after addition

 

[Dan_P]

Because of technology issues - I've had a couple days to think on the experiments, etc thus far, and I am going to change focus a bit - Because after looking at all the data - it is clear that (in this set up):

1. Rock taken from a tank (whether from a dark sump - or from a lit area of a display tank) - still has residual capacity to process ammonia over a week or so. I.e. the rock is not saturated with bacteria or ammonia consuming organisms.
2. Rock from a tank containing rock from a lit tank processes ammonia more quickly than rock from a dark sump.
3. The Rock from a lit tank does NOT process ammonia more quickly due to photosynthesis. EDIT - Though there may be differences in surface area on the rock in the formerly lit tank as well.
4. Multiple salt-water rinses with fresh water do NOT affect the ability of the rock to process ammonia
5. The glass, HOB filter, powerhead do not seem to be responsible for the increase in ammonia processing capability.
6. The 'Density'/'Porosity' of the rock MAY play a role in the differences seen between tanks in item 2 above - BUT - over time, the sump rock and lit rock both can process the same amount of ammonia over the same time period.
7. The current 'rock/water' is adequate to process 2 ppm ammonia in 24 hours repeatedly. Thus, meeting the criteria for Dr. Tim's 'cycle'. And thus - also in all likelihood the ratio of rock to water is similar to amounts in many reef tanks (about 1.5 lbs/gallon). Thus - at least in part - simulating a 'rip-clean'.
8. What amazes me is that despite multiple complete tank water changes - the water is crystal clear. No blooms, etc - and I mean - it is totally crystal clear. This was even when the light was on for multiple days in tank 2.
9. The Seachem alert badge continues to work 'appropriately'. The API tests - have not shown any 'false 0.25 ppm ammonia readings. In each case (1) - where I thought this was the case - 12 hours later it read 0.

So - now for the changes:

Todays experiment (#7) Protocol:

1. Empty tanks, wipe sides.
2. Scrub rock in tank water - for 5 minutes. Rinse rock with clean salt water Once
3. Add Rock and Clean water back to tank - do baseline measurements (because its a new batch of Saltwater)
4. Add 2 ppm ammonia to water - and do baseline measurements.
5. Measure ammonia as before

If 2 ppm is processed in 24 hours - will repeat the experiment again - to see if doing it more than once will affect processing capability (and also act as a replicate.

Thereafter - will skip RODI rinsing - and go directly to Tap water.

In the meantime - if anyone has any comments - suggestions - requests - feel free. I have not been able to obtain a low nitrite reader - but working on it @Lasse.

@Lasse, @Coxey81, @sixty_reefer, @Dan_P, @Garf, @brandon429, @Rmckoy, ETC ETC

Nicely done. I hope you have energy at the end of your study to write an article. The information that comes from you, @Coxey81, @LRT , etc. would be a wonderful resource. Maybe multiple authors would be a way to go.

Some observations and comparisons. I have not been able to establish a nitrifying biofilm on a glass or aragonite surface just by storing it in an unlit sump. I am not surprised your aquarium surfaces were duds.

When I do establish such a biofilm on a glass microscope slide with Bio-Spira, it has less capacity to oxidize ammonia than a glass slide coated with a single layer of aragonite sand. Is this because of a difference in surface area? Another possible reason why your surfaces appear to be duds.

Up to five minutes in a powerful vortex mixer in tap water or Instant Ocean did not affect ammonia oxidation of my Bio-Spira biofilm on aragonite sand a jot. It seems these bacteria biofilms are tough. The only time I killed such a biofilm was using a power drill set on high with a paint mixer attachment on a slurry of aquarium sand. I guess at some level of abrasion, a nitrifying biofilm can be damaged.

Looking forwards to the results of vigorous rock scrubbing. I will be moving from sand to aragonite rock slices to establish and study nitrifying biofilms. I will give them a good scrubbing to see if they are as tough as sand biofilms.

 

[MnFish1]

Nicely done. I hope you have energy at the end of your study to write an article. The information that comes from you, @Coxey81, @LRT , etc. would be a wonderful resource. Maybe multiple authors would be a way to go.

Some observations and comparisons. I have not been able to establish a nitrifying biofilm on a glass or aragonite surface just by storing it in an unlit sump. I am not surprised your aquarium surfaces were duds.

When I do establish such a biofilm on a glass microscope slide with Bio-Spira, it has less capacity to oxidize ammonia than a glass slide coated with a single layer of aragonite sand. Is this because of a difference in surface area? Another possible reason why your surfaces appear to be duds.

Up to five minutes in a powerful vortex mixer in tap water or Instant Ocean did not affect ammonia oxidation of my Bio-Spira biofilm on aragonite sand a jot. It seems these bacteria biofilms are tough. The only time I killed such a biofilm was using a power drill set on high with a paint mixer attachment on a slurry of aquarium sand. I guess at some level of abrasion, a nitrifying biofilm can be damaged.

Looking forwards to the results of vigorous rock scrubbing. I will be moving from sand to aragonite rock slices to establish and study nitrifying biofilms. I will give them a good scrubbing to see if they are as tough as sand biofilms.

we will see... honestly its getting old - the testing

 

[MnFish1]

EXPERIMENT 7 BASELINE

1. Tank 1 Sump rock, Dark, Rinsed scrubbed x about 2-3 minutes/rock (entire surface)
2. Tank 2 Tank Rock, Dark, Rinsed, scrubbed x 2-3 minutes on entire surface.
3. Both tanks drained, detritus removed
4. Rocks Rinsed with FSW then New FSW added to each tank - Baseline measures taken (Below)
5. 2 ppm ammonia added - Post ammonia pictures below.

Sump Rock




Tank Rock


Tanks Post rinsing

Tanks after ammonia

Ammonia Pre-addition

Ammonia after addition

EXPERIMENT 7 - Partial results:

So - at about 25 hours the Seachem alert showed 'safe' in both tanks. The API ammonia and nitrates suggest that tank 1 is lagging behind - so will check those tests again in 4 hours. BUT - in general - the scrubbing did not seem to show a big difference (with saltwater). with regards to ammonia processing capacity. Depending on the results later today - I will either scrub again today - with saltwater - using the same protocol as yesterday - OR wait until tomorrow. Note - with the pictures - the difference between tank 1 and tank 2 are subtle - but I think are still there. I.e. Tank 1 continues to process less ammonia.

BTW - in looking at the rock - in general - tank 2 contains more surface area - that is more difficult to scrub - its dead coral as well as rock. This could be causing the very slight difference. IDK. But - according to water volume and weight the 2 tanks are identical. This is just my observation. In any case - there is not a huge difference after scrubbing IMHO.

 

[Dan_P]

EXPERIMENT 7 - Partial results:

So - at about 25 hours the Seachem alert showed 'safe' in both tanks. The API ammonia and nitrates suggest that tank 1 is lagging behind - so will check those tests again in 4 hours. BUT - in general - the scrubbing did not seem to show a big difference (with saltwater). with regards to ammonia processing capacity. Depending on the results later today - I will either scrub again today - with saltwater - using the same protocol as yesterday - OR wait until tomorrow. Note - with the pictures - the difference between tank 1 and tank 2 are subtle - but I think are still there. I.e. Tank 1 continues to process less ammonia.

BTW - in looking at the rock - in general - tank 2 contains more surface area - that is more difficult to scrub - its dead coral as well as rock. This could be causing the very slight difference. IDK. But - according to water volume and weight the 2 tanks are identical. This is just my observation. In any case - there is not a huge difference after scrubbing IMHO.

Something I debate when optimizing the number of tests to conduct during an experiment: how few is enough to answer the question.

When thinking about the observations about the two different rocks, even though both met the criterion of so much ammonia processed in 24 hr, one could have done it in 12 hours, the other in 24 hr, making one actually better rather than the same. Maybe just pointing out the obvious

 

[MnFish1]

Something I debate when optimizing the number of tests to conduct during an experiment: how few is enough to answer the question.

When thinking about the observations about the two different rocks, even though both met the criterion of so much ammonia processed in 24 hr, one could have done it in 12 hours, the other in 24 hr, making one actually better rather than the same. Maybe just pointing out the obvious

Yes - you're exactly correct. I do check the Seachem alerts periodically through the day - and they seem to match - so if there is a difference - it seems like it would be small.

 

[MnFish1]

EXPERIMENT 7 - Partial results:

So - at about 25 hours the Seachem alert showed 'safe' in both tanks. The API ammonia and nitrates suggest that tank 1 is lagging behind - so will check those tests again in 4 hours. BUT - in general - the scrubbing did not seem to show a big difference (with saltwater). with regards to ammonia processing capacity. Depending on the results later today - I will either scrub again today - with saltwater - using the same protocol as yesterday - OR wait until tomorrow. Note - with the pictures - the difference between tank 1 and tank 2 are subtle - but I think are still there. I.e. Tank 1 continues to process less ammonia.

BTW - in looking at the rock - in general - tank 2 contains more surface area - that is more difficult to scrub - its dead coral as well as rock. This could be causing the very slight difference. IDK. But - according to water volume and weight the 2 tanks are identical. This is just my observation. In any case - there is not a huge difference after scrubbing IMHO.

Experiment 7 - Final results

After 27 hours both tanks were at 0 Ammonia on API - and 'Safe' on Seachem. As I said before - I might have been 'over-reading' Tank 1 Ammonia - both could have been 0 at 24 hours.

So - the conclusion - if anything - there is no affect rinsing and scrubbing the rock with salt water.
I will repeat Experiment 7 - to see if scrubbing a second day makes a difference. (with salt water)

 

[MnFish1]

PS - am going to try to time the start of the next experiment for later tonight - so I can do a 12 hour measurement. @Dan_P

 

[MnFish1]

@Dan_P. @brandon429. The hourly - or whatever measures with a seneye is what would be beneficial here.

 

[MnFish1]

Experiment 8. Replicate of Experiment 7.
2 ppm ammonia after scrubbing and rinsing rock.

Baseline:
EDIT (I accidentally deleted the Pre and post ammonia pictures) - they were 0 and 2ppm as before. Experiment Started at 1600

 

[MnFish1]

Experiment 8. Replicate of Experiment 7.
2 ppm ammonia after scrubbing and rinsing rock.

Baseline:
EDIT (I accidentally deleted the Pre and post ammonia pictures) - they were 0 and 2ppm as before. Experiment Started at 1600

FYI - at 12 hours (yes I got up at 4AM lol) - the tanks were both = on the Seachem alert - with them being somewhere between safe and alert - so it does not seem that one tank is processing ammonia at a 'faster' rate'. More updates later this afternoon. @Dan_P

 

[MnFish1]

Experiment 8 results

1. When I pulled the rocks out a day or so ago - they 'smelled'. before the first scrubbing - suggesting some 'stuff' was killed.
2. I really scrub this last time
3. Here are the results:

 

[MnFish1]

Experiment 8 results

1. When I pulled the rocks out a day or so ago - they 'smelled'. before the first scrubbing - suggesting some 'stuff' was killed.
2. I really scrub this last time
3. Here are the results:

This is more a lighting problem - the ammonia was definitely yellow

 

[MnFish1]

So the question for today is - try the tap water rinse? - or the RODI rinse - or another experiment - you guys tell me @Coxey81, @sixty_reefer, @brandon429, @Dan_P, @Garf, @Rmckoy, @Lasse - we now have 2 tanks that can process repeatedly - 2 ppm Ammonia.

 

[Dan_P]

So the question for today is - try the tap water rinse? - or the RODI rinse - or another experiment - you guys tell me @Coxey81, @sixty_reefer, @brandon429, @Dan_P, @Garf, @Rmckoy, @Lasse - we now have 2 tanks that can process repeatedly - 2 ppm Ammonia.

Here I come from left field: hydrogen peroxide or ChemiClean.

 

[MnFish1]

Here I come from left field: hydrogen peroxide or ChemiClean.

Great - well one comment was - chlorinated tap water rinse after scrub. But - if you wanted to use H2O2 - why? and what concentration - for how long? I mean we have 2 tanks of rocks behaving the same way thanks to @Lasse - saying 'keep both in the dark'

 

[Dan_P]

Great - well one comment was - chlorinated tap water rinse after scrub. But - if you wanted to use H2O2 - why? and what concentration - for how long? I mean we have 2 tanks of rocks behaving the same way thanks to @Lasse - saying 'keep both in the dark'

Both chemicals are used quite a bit and believed/hoped not to affect the nitrifyers. Your test bed might confirm the no harm conjecture. I think a popular dose of Hydrogen peroxide is 1 mL 3% in 10 gallons but @brandon429 is closer to this subject.