Brighteyes' Lab Notebook: Pony Mold*Update 5/31 Cleaning Results & Microscope

[BlueyBirdy]

This is awesome! I'm really excited to see how this progresses. The petri dishes are already looking so gnarly!

[BlushingBlue]

Wow, Majesty's plate is really something! The pink one reminds me of Fusarium, but I don't know why a pony would've picked that up.

I agree that the blue-green mold looks very much like Penicillium, but that's saying even less.
Very exciting stuff! I'm looking forward to what Phase Two brings!

[brighteyes]

Wow, Majesty's plate is really something! The pink one reminds me of Fusarium, but I don't know why a pony would've picked that up.

I agree that the blue-green mold looks very much like Penicillium, but that's saying even less.
Very exciting stuff! I'm looking forward to what Phase Two brings!

It does look like a Fusarium!  We have a Fusarium oxysproum in culture at my lab but it's definitely a deep purple, not pink.  I'm going to grow these in mold chambers so I can look at them under the microscope so we should be able to get better at IDing them after that.  It's possible the ponies picked up the Penicillin from the air in the lab.  Our air sampling plates are often replete with greenish molds (I assume are Pen. but who knows).  I always thought of Fusarium as more of a plant pathogenic mold so maybe the Fusarium is a transfer from some fruit.  I did the swabs under a BioSafety Cabinet and wore gloves but there is so much mold in this old building, anything is possible. 

So, next week I'll make a coctail of these molds and yeasts and swab our test subjects with it and store them away in sterile jars.  I'll pull the jars out every week or so and take and post photos.  Super fun science time

Thanks everyone for following along with the mad science project!

[Mirnyj]

Oh my goodness, this is so fascinating! I never thought of yeasts, what kind of effect they might have on ponies. It'll be interesting to see if your results will match the common consensus or if we'll learn something completely undreamed.

[Sweet_Stuff]

I'm so grossed out by all those photos...bleh.. but yet, I can't turn away! 

Same thing that happened to me in college when we did a fun experiment. The class (I think it was an environmental science class??) got one dish, swabbed anywhere they wanted to and waited to see if there was any growth. . . Mine was the worst by far.. I swabbed the classroom's outer door handle! It looked like some kind of yeast.??

Hehe...someone was nibbling some fruit!?! That penicillin looks so cool and gross..

[brighteyes]

Okay- I finally had time to inoculate the ponies today (wow that sounds really terrible!).

I vigorously washed all the plates of mold cultured from each pony with sterile saline and collected the wash in a sterile bottle.  The result was a yellowish, brownish nasty looking liquid filled with small mold chunks and hopefully many spores and hyphae:

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Prior to making the mix, I had boiled and soaked the test ponies in Oxyclean over night and then washed them thoroughly to *hopefully* remove oxyclean.  I then took photos of every angle of the pony test subjects so we can compare the existing spots and marks to any that develop or change during the course of the experiment.  All are available for examination in my photobucket folder Lab Notebook: http://s14.photobucket.com/user/brighteyespony/library/Lab%20Notebook

I did all the inoculations in a biosafety cabinet to reduce exposure of the pony samples to airborne mold and used gloves to reduce ponies' exposure to any molds or yeasts on my hands.  The general procedure was to put either 2mL (for Dollar General fakies) or 5mL (G1 ponies) of mold/yeast inoculum or sterile saline inside the pony and put the head back on and rotate the pony to ensure the inoculum reached all inside areas of the pony.  Some ponies were then surface inoculated by dipping a sterile swab into the mold/yeast inoculum and then swabbing the surface of the pony to distribute the culture.  Some ponies had additional sterile water placed in the jar to simulate a humid environment.  All ponies were then placed into a sterile jar and put in a cabinet in my lab to simulate a dark environment such as a basement. 

Here are the treatments I did:
The two Dollar General Fakies in each pack were inoculated with either 2mL sterile saline or 2mL mold mix and placed in sterile jars. No surface swab was done on the dollar general fakies.   Sterile water was also added to the jar to simulate a humid environment:

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Majesty had 5mL mold/yeast inoculum added and a surface swab with NO water to see if additional humidity makes a difference in mold growth.
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This Majesty had the exact same treatment as above except she had an additional 10mL sterile water added to the jar to increase humidity to differentiate the effect of added humidity on mold growth in ponies
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This Sundance had the same treatment as the above Majesty including 5mL inoculation, surface inoculation and 10mL water to increase humidity in jar.  She was used as a replicate for the general experiment.
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This Snuzzle had 5mL mix internal inoculation and 10mL water but no surface swab.  I did this treatment to see what effect mold growing inside the pony may have if no inoculation is made externally.  I.E. can mold growing inside a pony only make viable marks on the outside of the pony?
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[Sparkle Pony]

So excited to see the results!  I've never been this excited by mold before.

[kissthethunder]

This is fascinating!!! Please keep us posted!!

[slyons]

Nobel prize......NOBEL PRIZE!!!

[brighteyes]

Update in first post.  It really looks like we've got mold already. 

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The black dots were not present on the pony before inoculation. 

[Snapdragon]

AAAAAAAA!!! @_@ This is horrifying yet so interesting!! Keep us updated!! Poor ponies!

[Griffin]

How fascinating! I'll have to keep an eye on this thread.

[brighteyes]

Okay, so in a little over a week the mold growth has greatly increased.  I've got comparison photos showing the pony the day of inoculation (12/14) and the pony on Monday Jan 4th 2016.  Some of the molds are mimicking the halo effect we see with certain marks on ponies.  However, we don't know if the mold is only present on the surface and will be easily washed off or if it will invade or otherwise stain the vinyl and make marks like we often see on ponies.  I do know that I frequently get mold stains on test tube racks that have been left in the cold room too long and these can only be removed by soaking the rack in bleach.  So, I do think given enough time the mold will leave permanent marks.

Some of the marks look like raised pimples, as the one on Majesty shows.  It's difficult to tell from the photo, but it's definitely raised and wasn't there prior to inoculation. 

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Snuzzle is a pony that was only inoculated with mold inside and was not subject to surface swab.  The first visable mold growth I'm seeing is around her tail area and this raised bump.  Mold is also growing in the liquid at the bottom of the culture jar.

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The question now is, how long should I leave them before trying to clean off the mold to see if if left a mark?  What do you all think? A month, three months, six months, a year, five years?  I feel so bad for them- part of me wants to take them out now.  Once the experiment is over with, they will get completely cleaned and then customized as cool science-y versions of their original pony.  Majesty is going to look sweet with a labcoat and goggles

Here are the Majesty before and after photos for additional water.  I'll try and add the before and after without water tomorrow.

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Here is a close up of a colony that has a halo.  It's very difficult to take photos with through the glass. I hope you can see well enough.  If not I can try to pull the ponies out under the safety cabinet to get better photos.  It's similar to the halos we've seen, but perhaps more irregular.

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Other photos:

Spoiler

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[Sparkle Pony]

Hooooooly smokes!  This is just crazy, keep it up!  I'm not sure how long I would leave them though personally...

[Karebearcntdwn]

The mold probably won't stain, I think it might just be that the mold is in the pores of the plastic then leaches through the breakdown  in it. I would love to see a slide slice of one of these cultures under a microscope, it might get you closer to finding out the best way  to getting rid of the mold!