Sphagnan Experiment

[Mok]

I'm about to conduct an experiment with Sphagnan, which is a polysaccharide  commonly found in Sphagnum moss.  Some research has been conducted on this substance in the nordic countries, the first one that brought this substance to my attention involved putting zebrafish on moist sphagnum, an acetone-extract of sphagnum, and the purified sugar (I still don't know how they extracted it).  The control fish putrefied in less than two weeks, however the other three remained fresh for a much longer period of time.  It somehow inhibits bacterial activity via a highly reactive carbonyl group (it's somewhat unstable, too).  

Anyhow, I ground up some commercially available "green moss" (dried sphagnum) and am soaking it in acetone, which is slowly turning green as it extracts chlorophyll (and hopefully sphagnan) from the moss.  Once a randomly chosen point has been reached I will filter the moss and allow the acetone to evaporate, then collect the residue.  I will then extract the residue with warm distilled water and use this water to make MEA, which will be placed in two baby-food jars (I will also make two jars with regular MEA as a control). One sphagnan jar and one control jar will go into a pressure cooker at 15psi for 1 hour, and the other two will remain unsterilized.  

Once cooled and gelled, all jars will be inoculated with a speck of topsoil, covered with saran wrap, and incubated.  Photographs will be taken daily.

My expectations are that the control jars will quickly be overrun with growth while the cooked jar of sphagnan MEA will grow slightly less quickly,  while the uncooked jar of S-MEA will take the longest to show signs of growth.

If this experiment pans out I'll then attempt a similar experiment, innoculating the jars with unsterile edible mushroom prints.


I just figured I'd tell you guys, since it's mildly interesting in my opinion.

[Mok]

I'm done extracting the undetermined quantity of dried sphagnum moss.  I dried the acetone outside on a pan with a heating pad underneath it.  It evaporated quickly even though it's winter.

Anyhow, what was left was a thick lime green liquid (most likely water) that smelled very much like fish body oil.  Yecchh.  I took this up in a small quantity of Deer Park brand drinking water fresh from the bottle.  I'm fairly sure the lime green component is suspended chlorophyll, I'm betting it'll settle overnight.  

I may try extracting it with some kind of oil after I experiment with it a bit, to see if I can get rid of that nasty smell and color.  Anyhow, next step is the microbial tests, which I'll conduct tomorrow.  Night world!

[Mok]

Ok

4.5g agar
5g light malt extract

added to

240ml boiled Deer Park bottled water

Stirred, cooled slightly, then the 10ml or so of lime-green sphagnum water was added and stirred more.

(The green part did not settle out)

Poured enough to cover the bottom of a 1oz babyfood jar, they're now cooling on top of the microwave.

Can't wait to inoculate!

[Mok]

Ok, I deviated from my original experiment plan.  Perhaps at a later date I'll come back to it.  Anyhow, here's the update:

I deposited a small chunk of dried Pleurotus tuberregium sclerotia into the center of one of the jars on Jan. 7 (it was given to me by an old friend over 5 years ago and has been in storage since then).  This chunk had been amateurishly rehydrated by holding it between my fingers and placing it under a running tap.  Today, Jan. 10 I have observed a growing ring of mycelium surrounding the chunk and fluffy white fibers on the chunk itself.  No other growth has been observed in this jar.


The control jar is showing no growth, nor is the jar that has been inoculated from a 4 year old unsterile agaricus print.  We will have to wait on those, I have been told that dried spores can take up to 2 weeks to sprout.  I should have rehydrated them... oh well.

I would take pictures but I haven't figured out how to make them nice and clear.

Science, ho!

[Mok]

The sclerotia chunk appears to be burrowing into the agar.  It is covered in a clear gel, there is about a 2mm ring around the gelatinized chunk that rises up almost 1mm, it is covered in what appear to be fine white fibers, although not numerous enough to appear as fur.  It appears to be burrowing into the agar.  A thin ring of textured surface surrounds it, I can't make out fibers but I'm guessing that's the advance guard.  No other growths can be observed.

I have replaced the baby-food-jar lid with a single layer of saran film for more satisfying observation.

Status unchanged on jars B and C.

I wonder if the sphagnum prevents spores from sprouting and/or single-celled organisms proper operation.

[Mok]

I took the jar out today and was shocked to discover the mycelium glowing fiercely!



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Just kidding, trick of the light.


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Sorry about the crappy image quality, I'm a terrible photographer.

[Mok]

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it would be really cool if it grew to occupy the entire jar.


I think the next step will be to extract more sphagnan, then coat a bunch of cooked rye grains with it, then inoculate with a chunk of this.  

Science, ho!

[Mok]

The motherjar is infected!  Quite stinky, too.



This jar is what I mixed the agar in and poured from, it was sealed the day it was made and hasn't been opened (or looked at) since.  "kewl" in chat observed that it could be anaerobic bacteria.  The aforementioned fuzzy was aerated for several minutes when the first pictures were taken, that's how I got the close-up (with my dis-assembled webcam).

The control jar and the sporulating jar are both infected with stinky bacteria.  I'll let it grow out so we can see what it is.  (Neither has been opened since poured)

[Mok]

Alas, my experiment has failed.  The mycelium has succumbed to the deadly grey spiderweb as legions of yeast surrounded it.  

1. The sphagnan may have been destroyed by the heat from the agar.

2. The sphagnan may have been destroyed during the processing of the product obtained.

3. The contaminants were specifically immune to sphagnan's effects.

4. The ability of sphagnan to prevent decay may be limited to specific physical configurations, excluding those created for this experiment.

5. The moss utilized did not contain sphagnan to begin with, being that it was a different variety of sphagnum.

6.  The moss was not sphagnum at all, but one with similar appearance.


I may have to ask the norweigan researcher to supply me with the pure substance.

I bet he'd do it.