What is this? Contam?

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bambalez
Fellow

Registered: 10/23/18
Posts: 6
Loc: Portugal

What is this? Contam?

#25559819 - 10/23/18 07:16 AM (7 years, 9 months ago)

Hi guys,
first and foremost i'd like to thank anyone who took their time to see this post and maybe help me out!

I've recently started working with agar and since i lack experience with it some questions came up, even tho i've tried to search answer for these.

After a few inoculated plates of agar i noticed this pattern in the growth of mycelium. Most parts of it grow white and fluffy, as i would say an healthy mycelium. But some other parts have this slightly yellow discoloration which almost blends in with the white. I can´t tell if its contamination, although i've had some, rarely, and they were quite obvious.
Anyways, i will further explain the process involved in inoculating this such as procedures and recipes, and also add a few photos from various perspectives, from the plates.

For the agar recipe i used potato dextrose agar (PDA) powder from Biolife with added peptone (also from Biolife) in the following ratio measures:

-1000 Destilled water
-42g PDA (5g potato extract, 20g glucose, 17g agar)
-0,5% peptone

The solution was put inside a glass bottle and closed with a metal lid containing a hole, micropore tape on it and aluminium foil. the bottle was sterilized in a preassure cooker at 15 psi for 25 minutes and let cool down.

Disposible gamma-radiation sterilized petri dishes were used for this.
The agar solution was poured into the dished inside a totally closed still air box (glued with sleeves and gloves) which was thoroughly clean with 1:1 water:bleach solution before hand. Also, before pouring i the agar i sprayed the bleach solution and waited to let air agents inside the box settle to the floor.

After solidifying i inoculated 2 agar plates with different strains from spore syrynges by dripping 2 drops in the middle of the agar and spreading them with a sterilized disposible inoculation loop.
One other plate was used to introduce growing mycelium from other plate which presented the same characteristic as these do now (i thought that if it was contamination then maybe i could the the sample by removing a small healthy part of the previous agar plate and putting it in a new one)

Plates were sealed with parafilm and put into a 27Cº chamber.

I think i covered most of whats relevant to this topic. But please feel free to ask for any information i might have missed.
I'd like to add also that spore syringes strains used were Cyanescens and golden teacher, and the transfer from agar to agar carried the mazapatec strain.

Again, thank you for taking your time to read and maybe help out.
I wish you all the best!

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