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InvisibleHyphai
Registered: 04/03/14
Posts: 517
Bacteria in Spore Syringe.
    #20346667 - 07/29/14 12:56 PM (10 years, 5 months ago)

So I have some spore syringes from a reputable source. Lately I've been using nothing but prints, but occasionally I will use spore syringes to start something new off.

When I was first starting off I never used agar, just inoculated grain from the getgo and never had much contamination. Since then I've moved to agar and noticed that whenever I inoculate a plate from spore syringe there is a much higher rate of contamination, often as a splotch in the very center. Since I never ran into this issue when going straight to grain, I am starting to assume that this bacteria was always there and the mycelium was somehow able to outrun it?

So basically the question is, is there almost always bacteria in these syringes, or have I just been getting some bad luck with acquiring spore syringes?

Edited by Hyphai (08/15/14 07:38 AM)

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Invisiblemushmagic
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Re: Is This to be Expected? [Re: Hyphai]
    #20346687 - 07/29/14 12:59 PM (10 years, 5 months ago)

How much spore solution are you dropping into each plate? You only want a like a drop or so. If you have prints you could just swipe that on the plate.

Either way just transfer away from the bacteria.


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Edited by mushmagic (07/29/14 01:00 PM)

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OfflinePsiloBeast
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Re: Is This to be Expected? [Re: Hyphai]
    #20346692 - 07/29/14 01:01 PM (10 years, 5 months ago)

Well if you haven't had problems in the past I'd chalk it up to contam during inoculation of your agar OR the agar itself is contaminated. Are you flaming your needle red? Working in a SAB?


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InvisibleHyphai
Registered: 04/03/14
Posts: 517
Re: Is This to be Expected? [Re: mushmagic]
    #20346696 - 07/29/14 01:01 PM (10 years, 5 months ago)

I am only using a drop, or two if the second one sneaks out with the first. It is always bacteria also.

I've also done many tests. I will inoculate say 3 plates with syringe A, and 3 plates with Syringe B. All 3 of syringe A will contaminate, but B will be just fine. I am pretty confident in my technique, so I will assume that this is the fault of the syringes themselves.

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Invisiblemushmagic
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Re: Is This to be Expected? [Re: Hyphai]
    #20346705 - 07/29/14 01:03 PM (10 years, 5 months ago)

Yea it's likely the inoculant then.


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InvisibleHyphai
Registered: 04/03/14
Posts: 517
Re: Is This to be Expected? [Re: PsiloBeast]
    #20346716 - 07/29/14 01:06 PM (10 years, 5 months ago)

Quote:

PsiloBeast said:
Well if you haven't had problems in the past I'd chalk it up to contam during inoculation of your agar OR the agar itself is contaminated. Are you flaming your needle red? Working in a SAB?




I'm using a flow-hood and a bacti cinerator. Lysol before and after each use and every surface is wiped with iso-propyl alcohol. I use gloves and tyvek sleeves and wipe my hands down with alcohol religiously.

Again, I am certain the syringes are contaminated. Spore prints have never given me this issue, (though molds have appeared right on the streak pattern, i chalk that up to a dirty print cause it has only happened with one print). Transfers have never given me an issue as well. It's only spore syringes, yet I never had contamination's going straight to grain as a beginner. Then again I was nowhere near as active in the hobby as I am now.

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Invisiblemushmagic
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Re: Is This to be Expected? [Re: Hyphai]
    #20346723 - 07/29/14 01:08 PM (10 years, 5 months ago)

Does the bacteria begin to appear right where the site of inoculation is?


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InvisibleHyphai
Registered: 04/03/14
Posts: 517
Re: Is This to be Expected? [Re: mushmagic]
    #20346730 - 07/29/14 01:10 PM (10 years, 5 months ago)

Yep right in the middle. One time within 24 hours, but most of the time I can see it after 48. I really wonder if these syringes would work going to grain though.

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Invisiblemushmagic
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Re: Is This to be Expected? [Re: Hyphai]
    #20346748 - 07/29/14 01:14 PM (10 years, 5 months ago)

Is it maybe always the same syringe re-used that the bacteria results from?

EDIT: Like possibly a compromised syringe?


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Edited by mushmagic (07/29/14 01:15 PM)

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OfflinePsiloBeast
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Re: Is This to be Expected? [Re: Hyphai]
    #20346754 - 07/29/14 01:15 PM (10 years, 5 months ago)

It going to grain and working makes me wonder if it's not the agar...

I've had tons of (self made) spore syringes work fine. No contams. I'm just wondering if you're having contam issues from your reputable spore vendor...hmm. Don't know man :shrug: hope you find out though.


--------------------
Whosoever is delighted in solitude is either a wild animal or a god...
                                    -Aristotle
              A COMPLETE GROW LOG (ATL# 7)
                          BEAST STYLE LINKS
Water Extraction Tek,Frank's Monotub Tek ,Do What He Does...

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Invisiblemushmagic
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Re: Is This to be Expected? [Re: PsiloBeast]
    #20346762 - 07/29/14 01:16 PM (10 years, 5 months ago)

Wait so you have tried the syringe on grain OP?


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InvisibleHyphai
Registered: 04/03/14
Posts: 517
Re: Is This to be Expected? [Re: PsiloBeast]
    #20346769 - 07/29/14 01:17 PM (10 years, 5 months ago)

Well I don't know if it would work. I don't really want to waste any more resources with these few suspect syringes.

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InvisibleHyphai
Registered: 04/03/14
Posts: 517
Re: Is This to be Expected? [Re: mushmagic]
    #20346776 - 07/29/14 01:18 PM (10 years, 5 months ago)

Quote:

mushmagic said:
Wait so you have tried the syringe on grain OP?




Well I have one grain jar ready to go with a SHIP and all that so I could test it out on and see what happens :shrug:.

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OfflinePsiloBeast
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Re: Is This to be Expected? [Re: Hyphai]
    #20346793 - 07/29/14 01:22 PM (10 years, 5 months ago)

Well good luck man.

Try some more on agar too if you've got extra.


--------------------
Whosoever is delighted in solitude is either a wild animal or a god...
                                    -Aristotle
              A COMPLETE GROW LOG (ATL# 7)
                          BEAST STYLE LINKS
Water Extraction Tek,Frank's Monotub Tek ,Do What He Does...

Extras: Filter Print Post Top
Invisiblemushmagic
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Re: Is This to be Expected? [Re: Hyphai]
    #20346795 - 07/29/14 01:23 PM (10 years, 5 months ago)

It's worth a shot to help you get an idea. You can always make more grain.


--------------------


Trade list in journal (partially under construction; more to be added)


Don't judge a man by what kinda shoes he in, judge a man on where that man's shoes been.

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InvisibleHyphai
Registered: 04/03/14
Posts: 517
Re: Is This to be Expected? [Re: mushmagic]
    #20410692 - 08/12/14 09:03 PM (10 years, 5 months ago)

Just got around to testing it on grain, I'll update in a few days.

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InvisibleHyphai
Registered: 04/03/14
Posts: 517
Re: Is This to be Expected? [Re: Hyphai]
    #20421357 - 08/15/14 07:27 AM (10 years, 5 months ago)

I see definite germination and no sign at all of bacteria, something I never got to see when I tested it on agar. So I'm assuming that the mycelium is able to outrun the contaminant when going straight to grain. Cool. That means I can use this PE syringe :thumbup:.

Edited by Hyphai (08/15/14 07:40 AM)

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InvisibleSwampEyes
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Re: Is This to be Expected? [Re: Hyphai]
    #20421391 - 08/15/14 07:43 AM (10 years, 5 months ago)

the liquid in the syringe could possibly be incubating the bacteria on agar moreso than on the grains since the liquid just sits there when you put it on agar as opposed to dispersing amongst the grains. you could always put one of those colonized grains to agar and isolate from there to get a cleaner PE culture


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OfflineQuexl
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Re: Is This to be Expected? [Re: Hyphai]
    #20422066 - 08/15/14 10:43 AM (10 years, 5 months ago)

Grain prep decides if grain goes bacterial more than the syringe does. Even perfectly prepared pressure cooked grain has bacteria spores all over it, but when it's dry enough its abiotic and doesn't grow. Sounds like you did that right :thumbup: nice.


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InvisibleHyphai
Registered: 04/03/14
Posts: 517
Re: Is This to be Expected? [Re: Quexl]
    #20422811 - 08/15/14 01:29 PM (10 years, 5 months ago)

Very cool. I hope this works out.

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