Bacteria in a bottle, Myth or Fact

[Brew12]

Not sure if this question is answered anywhere in this thread, as I did not read all of the posts, but have you viewed any of the bacteria under a microscope? I'm not sure of the size of the particular bacteria used in these bottles, but I would like to do a lab in class viewing them if our microscopes are capable of doing so. I tried finding someone online with this, but most results were from a research lab rather than a high school/college course, and had images from Electron Microscopes. I wasn't sure if this was due to the size of the bacteria requiring the more powerful microscope, or if there was just a lack of interest at a lower level to view these bacteria.

Ok, got some pictures for you. I won't claim to be an expert with a scope.

Biospira at 100x magnification Lots of bacteria and other "stuff"

Taking it up to 250x gets us this

And at 1000x we get this:

 

[brandon429]

oil immersion microscopy req'd

we had to use leica $1500 scopes to see bac colonies if im not mistaken the resolution was 500 or a thousand x, 100 will not do even for colonies unless 1998 recollection is far off, its been a while but those pics made me recall how hard it was to place that drop, drag that slide, twist the resolution wheel within a micrometer of what it takes to go from totally blurry to a true bacteria/colony ~ it was hard work to get there.
I remember the prof directly telling me oil immersion was required to see all manner of mixed and generalized aerobic bacteria
.


are we to discern those might not be pure strain solutions in tow, what kind of solids or precipitates or algae or actual reef diatoms can be in the bottle after the production/purification process?

that picture is fascinating from a non bacteria perspective 110% I believe your photograph shows proof these are not isolated pure strains of bacteria because we're looking at substrates that wouldn't be sterile upon addition to that mix. those are cells of living animals or plants in some way, can't wait to hear from the retailers following along.

 

[CDavmd]

Yes as stated by Brandon ...bacteria require high magnification with a good scope. At least 1000x and an oil immersion lens. Even at those powers they are difficult to see without a gram stain.

 

[Mr.Asbury]

Thanks for the information, everyone. Gram staining wouldn't be a problem, but I was hoping to avoid the oil immersion scopes that we have - they're pretty much only used by the AP level kids and are, like you said, very expensive. I know we've looked at strep & staph under our scopes, so I was hoping the nitrifying bacteria were as large as they are. I don't want to risk my students that aren't as familiar with using the scopes to damage them from trying to turn the adjustment knobs past their limit - we have issues with this fairly regularly in the general level sophomore classes where all students use the scopes. Our general ones are a little less expensive, and we usually have to have someone come in every other year to fix them all up.

I'll just save microscope work for when we look at creek/pond water & look at some of the protists there - they're a lot larger and easier to see. I had already planned on looking at inverts from our creek with dissecting scopes, so I can go with our regular light microscopes at that point too. I believe they're 430x max, so there's almost no chance of seeing the bacteria if you're saying they require 1000x.

Thanks again for taking the time to check!

 

[Dr. Reef]

Okay guys I just now got home and at about 4 a.m. would have been the 24 hour mark but it's about 8 a.m. when I'll be posting the results. I know I'm 4 hours late but I'm sure four hours is not that big of a deal to show a whole lot of difference. Anyways stay tuned.

 

[Dr. Reef]

I tested all 5 tanks. As of 8am central time. (28 hrs after dosing bottle bacteria)
at 12 hrs mark i had seen no change in color or reduction in any of the tanks. All 5 showed 2 ppm.
At 28hr mark i have a conclusive result. I am uploading the video at youtube so everyone can see the test as it progressed.
As soon as its up i will post the results along with the video.

 

[Dr. Reef]

At 28 hrs after dosing bacteria (at strength manufacturer recommend) i have conclusive result.

1. Control tank is still 2 ppm.

2. Seed, Prodibio and Dr Tim products were able to pull ammonia down to about half way at 1 ppm.

3. While Fritz Turbo Start seems to have cut the 2 ppm ammonia down to 0.5 ppm.


From left to right
Seed, Prodibio, Dr Tim, Fritz

 

[Dr. Reef]

 

[Dr. Reef]

For better results and second opinion i am testing all samples with Red Sea kit as well. stay tuned.

 

[4FordFamily]

Wow impressive! Good work boss!

 

[Dr. Reef]

Halfway through Red Sea ammonia test (takes 15 min) (at 7min)
I can already see the same results developing like API kit.

From left to right
Dr Tim , Fritz turbo start 900

 

[Dr. Reef]

Final results on Red sea confirms reduction from 2ppm to 1.2ppm on Seed, Prodibio and Dr Tim.
While Fritz show darker than 0.4 but lighter than 0.8ppm

Dr. Tim (right at 1.2 ppm)
Fritz Turbo Start 900 shows about 0.5 ppm

 

[DieHardPhotog-Reefer]

Final results on Red sea confirms reduction from 2ppm to 1.2ppm on Seed, Prodibio and Dr Tim.
While Fritz show darker than 0.4 but lighter than 0.8ppm

Thanks for taking all of this effort! Greatly appreciated. And thanks to all the vendors who have shared in the journey too!

 

[Dr. Reef]

Sorry guys in this pic. Red Sea test for ammonia at 28 hrs after dosing bacteria.
I was so focused on posting results and uploading videos i forgot to take a pic of all samples at 15 min mark and let it sit longer than 25 min.
Anyways i observed
at 15 min Seed Prodibio Dr Tim were all at 1.2'ish
while Fritz was 0.5-0.6ppm

Here is a pic after 25 min but still you can clearly see first 3 to be around 1.2-2 ppm while last vial Fritz to be much lighter than other at about 0.5ppm

 

[Dr. Reef]

On with the study, Seachem Seed requires daily dosing so i dosed bacteria again today.
Will continue to test at 24 hr mark for next few days. After a tank reach 0 i will dose 0.5ppm ammonia daily and continue to monitor making sure ammonia does not return.

 

[Brew12]

Great work! It's nice when results are repeatable!

 

[Dr. Reef]

Yes sir, will test for next 5-7 days then bleach and reset again.
Going to run this 2 ppm study total of 3 times before i call out a better product, So far Fritz is looking good.
Once done with this then Study #3 will start for 3 rounds, that study will include products that didnt make it on this test.
I am excited to announce Bio Spira and MicrobeLift has sent their samples while ATM colony has not responded and Fluval Cycle flat out refused to provide sample and did not respond to me buying a fresh bottle from them. Shame.....

 

[Dr. Reef]

I think evaporation that we noticed last time and what i am seeing this time is completely random. Even though all tanks are setup in same area and 6" apart from eachother, there is no set pattern neither last time nor this time and it seems all random how and which tanks evaporate more or less.
Here is a pic as of today

 

[Dr. Reef]

Ok guys at 48 hrs i have the final results.
Fritz turbo start 900 hit 0ppm.
Others still seem to be stuck at almost 2 ppm. Hard to say on API kit but its definitely more closer to 2 than 1ppm. I will give it a few more days to see its bacteria starts to colonize. If not i will bleach and reset again for round 2 of same test.

Here is a video and pics of the testing as of today (48hr after dosing bacteria)

 

[Dr. Reef]

Evaporation seems to be in a random but similar pattern in last 2 days. Almost kicking on and off within same hours of the day but which tank goes first is still random.