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OfflineNotJonHamm
Stranger Danger

Registered: 11/14/17
Posts: 73
Loc: Canada Flag
Last seen: 5 years, 8 months
Agar Contamination or ... ?
    #24885164 - 01/02/18 02:28 PM (7 years, 15 days ago)

Hi Shroomery! I've been lurking for about a couple months now, reading as much as I can, and learning from everyone's experience first by becoming somewhat adept at using the search function. After researching, I decided to start with agar to minimize contamination and also have a pretty neat hobby. So I guess I'd like to thank everyone who has taken the time to write up teks and offer advice to the uninitiated; it's pretty impressive how big and supportive this community is! But...

However, I now find myself scratching my head after I have went from a multispore syringe to agar, I'm not having results indicative of successful inoculation that I have seen in the many agar plates I've seen on this forum. The first attempt I ended up with a lot of pink growth as shown by these plates with a PDA agar:

This growth was present on 20/30 plates, so I went back to the drawing board to up my sterile technique. After my infrared thermometer arrived, I also noticed that during my 12 hour light-on cycle, the plates were getting to upwards of 85degF, which I assume compounded any problems.
I then poured a new set of plates, this time with a MEA agar to increase clarity to better evaluate growth and changed my grow room setup so that during the light-on cycle, my max temperatures are roughly ~78degF.

When I poured/inoculated my new plates, I also made a set of control plates where I went through the motions of inoculating with a syringe, but without pushing the plunger to evaluate my technique. Over the last week, there has been zero growth of any kind on the control plates, so I feel comfortable that my sterile technique is good. Now though, I'm getting an altogether different growth types on the plates I did inoculate. The following examples are pretty representative of all 20 plates I inoculated a week ago:
- Some plates have white growths, but it appears to be neither tomatose or rhizomorphic. Just wispy white streaks going hither and thither.

- Other plates are also showing white growth, but with more solid sections (at roughly 12 o'clock on both images).

- Others have yellowish dots or areas that which once were white circular growths turning blue in color.

Just for a reference, this is representative of how all the plates looked after a day or two:

On AUS-MS-004-000-AA (3rd row, 4th column) at 12 o'clock, you can also get a pretty good look at the "thicker" white growth.

So I guess my main question is, are any of those plates indicative of mycelial growth? If so, should I try and start making some transfers to new agar dishes to isolate healthy growth from contaminated areas?
Secondly, if this is all just different forms of contamination from the MS syringes I bought, should I just throw them out and buy some new spores from a more reputable seller?
Thanks in advance for reading all this and sharing any information/experience you might have with this.

Edited by NotJonHamm (01/09/18 10:23 PM)

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OfflineShrooms4life
TrippyShaman
Male


Registered: 06/08/17
Posts: 1,376
Loc: Under the Viel Flag
Last seen: 30 days, 19 hours
Re: Agar Contamination or ... ? [Re: NotJonHamm]
    #24885169 - 01/02/18 02:32 PM (7 years, 15 days ago)

I haven't gotten into agar with practice yet but I've started doing my research because I'm about to get started and I've yet to see any growth like yours are showing, idk if it's contam or just shitty spore syringe but one of the greats will jump in here to help you soon! Best of luck brother.

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InvisiblebodhisattaMDiscordReddit
Smurf real estate agent
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Registered: 04/30/13
Posts: 61,915
Loc: Milky way
Trusted Cultivator
Re: Agar Contamination or ... ? [Re: NotJonHamm]
    #24885194 - 01/02/18 02:46 PM (7 years, 15 days ago)

Looks like you got a bad syringe. Send me a PM where you got it from.
Your agar work looks good. Those odds are more than bad technique. Besides your controls not going bad.

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OfflineShrooms4life
TrippyShaman
Male


Registered: 06/08/17
Posts: 1,376
Loc: Under the Viel Flag
Last seen: 30 days, 19 hours
Re: Agar Contamination or ... ? [Re: bodhisatta]
    #24885325 - 01/02/18 03:47 PM (7 years, 15 days ago)

Damn i feel like I'm getting good at this *grinslikehedroppeda10strip*

Thanks for the confirmation bod shit looked weird and the op seems like a very knowledgeable guy to me.

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OfflineNotJonHamm
Stranger Danger

Registered: 11/14/17
Posts: 73
Loc: Canada Flag
Last seen: 5 years, 8 months
Re: Agar Contamination or ... ? [Re: Shrooms4life]
    #24885392 - 01/02/18 04:13 PM (7 years, 15 days ago)

Yeah I took a chance on a bad reviewed supplier in favour of low prices and free shipping, but it seems to have bit me. So I think I'm going to suck it up and purchase from a sponsor to avoid wasting plates and more importantly, time.

Thanks for taking a look though, guys. I was optimistic that I would have some mycelium, but alas, success will come another day!

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OfflineNotJonHamm
Stranger Danger

Registered: 11/14/17
Posts: 73
Loc: Canada Flag
Last seen: 5 years, 8 months
Re: Agar Contamination or ... ? [Re: NotJonHamm]
    #24902036 - 01/09/18 10:18 PM (7 years, 8 days ago)

Okay, so I've given it another shot! I bought some spores from a dependable sponsor that arrived after a couple days and then made up some new plates. I got some PESA, Ecuadorian, Chitwan Nepal, and Treasure Coast. So far except for a few spots on two plates of the Ecuadorian, there hasn't been any of that white slimy growth (bacteria?). As you can see from the pictures there's a couple big slices taken out of two plates in a hail Mary attempt to save them from the slime.
So I'm still waiting on something to grown on the PESA, Chitwan, and Treasure Coast plates, but I have growth on 5/6 of my Ecuadorian plates! And dare I say, what I think is some actual non-contamination growth (hopefully)! What do you guys think?

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