Bacterial Analysis.

[ShaunW]

Hi, I am looking for one/two specifically established reef tank(s) that I would like to analyze the bacterial populations within. This would be a pilot study, toward a much greater future in depth analysis, including skim-mate analysis. I am presently doing the analysis on my own tank, however, any result is uninterpretable, since my sample number is one (acts of GOD can happen with just one sample).

The reef tank(s) in question would have to have a few criteria met.
1. More than one year old, preferably 3-4 years old.
2. Unchanged over time, except for growth.
3. Growing and healthy, no large corals deaths in the last 9 months.

In terms of husbandry, i.e. DSB, BB, SSB, zeovit, etc, it doesn't matter for this pilot.

What is required:
Water samples from the water column (50mL) daily for five days taken at the same time. Then twice weekly samples, one during lights on and the other 1-2 hours after lights out. Two samples a day, two times a week for two weeks.

I would provide all the tubes to collect samples, and you would have to meet me to give them back.

 

[cali_reef]

I'll do it and I can provide water from two independent tanks, one is about 2.5 years old(mostly fish, very little coral) and the other full blown SPS reef is about 1.5 years old.

 

[prattreef]

I'm in too. My 125 is nearly 10 years old ( same rock and sand) and the bowfront is approaching 4 years. RD

 

[ShaunW]

DONE! Thanks guys!

Randy, providing you tubes would be easy, since I could just swing by Pratt.

Pierce, I would make the drive to visit you. Let's work out a date/time in the next couple of weeks.

THANKS GUYS, .
Shaun.

 

[ulmo]

That would be a very interesting study. It'll be great to see what types of bacteria are isolated. What media are you culturing these bacteria on?

 

[prattreef]

Maybe you could swing by pratt and we can both visit Pierce

 

[BroRage]

..

has it ever been known for fungal infections to come from our fish tanks...for example ring worms and stuff like that...

 

[Deanos]

has it ever been known for fungal infections to come from our fish tanks...for example ring worms and stuff like that...

Yes. A once active member was forced to quit the hobby after being infected by his tank. :shhh:

 

[spykes]

shuan will you be showing any pictures of culture plates or media? i like to see the works in action =D

 

[Josh]

Sounds like a cool project.

 

[ShaunW]

That would be a very interesting study. It'll be great to see what types of bacteria are isolated. What media are you culturing these bacteria on?

Hi Peter, ! Any input you would like to provide would be extremely helpful in terms of experimentation, interpretation, etc.

My first goal is to achieve a quantitative analysis of the amount of bacteria present per mL of tank water in each tank tested. Please keep in mind that I will be measuring planktonic (free swimming) bacterial populations. Biofilms, adherent/attached bacterial numbers will be left for another day.

It is well documented how many bacteria per mL are present on natural reefs. This free swimming, planktonic bacterial biomass represents an important part of the food web. Many of the species of bacteria are uncultureable due to their specific nutrient requirements, etc, or lack thereof. So to begin, I am going to use a bacterial cell counter to measure bacterial counts from mine, Randy's and Pierce's tanks. This will allow me to include many of the uncultureable bacteria in the counts and will give me a N=5 value such that deviations between different tanks will be seen.

After this part is done, with the help from Christine, we are going to isolate and characterize some of the species present.

Finally skimmate analysis will occur.

 

[ShaunW]

Maybe you could swing by pratt and we can both visit Pierce

Sounds like a good plan to me. Weekends would work best for a road trip, .

 

[ShaunW]

shuan will you be showing any pictures of culture plates or media? i like to see the works in action =D

Of course Dave, :iamwithst ! What's a scientific study without pretty pictures!

I plan on doing this with the same rigor that I would perform in my professional microbiological career, with all the nessesary controls and bells and whistles.

 

[spykes]

Of course Dave, :iamwithst ! What's a scientific study without pretty pictures!

I plan on doing this with the same rigor that I would perform in my professional microbiological career, with all the nessesary controls and bells and whistles.

are we gonna see any pics of DNA fingerprinting? (electrophoresis pics) sounds like fun stuff. i wonder what kinda taxomy you'll find. This would be awsome for me because this year im learning this stuff.

 

[cali_reef]

Weekends are mostly good for visits, or I can also bring them to work in midtown.

 

[Spracklcat]

Hi guys,

First Shaun I have those media recipes for you--found a few more as well so I can get them to you soon.

Pierce and everyone--thanks for the help! This is going to be good actually. Tag team labs

Christine

 

[ShaunW]

Hi guys,

First Shaun I have those media recipes for you--found a few more as well so I can get them to you soon.

Pierce and everyone--thanks for the help! This is going to be good actually. Tag team labs

Christine

Thanks Christine. That would be great.

So here are the six tanks to be analyzed:

Shaun - mixed reef, 80% SPS system, 10 years old.
Randy - 125 gallon, 10 years old (same rock and sand) and the bowfront approaching 4 years.
Pierce - two tanks, 2.5 years old (mostly fish, very little coral), and the other full blown SPS reef 1.5 years old.
Kedd - Mixed Reef that is mostly SPS.

 

[Spracklcat]

Shaun,

I have two of mine that might be good for comparison--a 29g mixed reef (LPS, SPS, softie, fish) that's been moved a bunch, last move 5 months ago (call this the "instability tank" ) and a 15g macroalgae/fish tank with refugium (I know it's bizarre, but this is a holding tank for two fish of my brother's that don't play nicely with the other fish and have to be isolated).

Also, I have sample tubes and everyhting all set up that I can mail out and have mailed back if we want to go that route--some are plain, some have some marine media (set up like human culture tubes)

 

[ulmo]

Shaun,

I know that it is very difficult or nearly impossible to culture some of the marine species of bacteria. In the past, I have tried using sea water with the addition of agar as a culture media but with limited success. I did manage to grow some ocean psychrophiles(cold-loving bacteria). The one source I can recommend, if you haven't check already, is the Difco media guide. They have some selective media for culturing marine bacteria. Good luck with the research

 

[Spracklcat]

Hi guys,

With regard to fungal infections from marine, like ringworm: I have never heard of, nor can I find reference to, people contracting fungal disease from an aquarium. Bacterial infection, absolutely YES and I urge everyone to wear gloves when handling animals and to avoid sticking your hands in your tank if you have open cuts (and of COURSE I always follow my own advice....um, well....yeah). I've isolated Shewanella putrefaciens and Shewanella algae from home aquaria, both of which have been known to cause infection in humans when given a portal of entry into the body. Additionally I know of more than one aquarist that has been hospitalized after just handling fragged anemones or corals--both cases doctors never isolated a causative pathogen (not surprising) and attributed the illness to toxins, but I am not so sure there wasn't a bacterial component. And there weren't quickie illnesses either--these were several day hospitalizations.

Media: Difco makes a dry "Marine agar" and a broth version, both of which I have used and like. They are not minimal nutrient media though, so I don't think we will be able to grow all of the pelagic bacteria on those. I am going to try making a low nutrient media, something like R2A, but with salt water instead of distilled. We'll see if that pulls out more/different species.

One other thing I will be looking at (with Shaun? ) is the comparison of species on coral surfaces to that in the water, and to that on other species. These species will grow more easily in typical nutritive media because the coral surface, being mucus-covered, has a lot more food and harbors more heterotrophs.

There are also quite a few recipes for media for specific marine species we can try. I'll forward them along to anyone who is interested.