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Offlinecosmicjam
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Registered: 02/05/11
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Last seen: 12 years, 29 days
Persistent bacterial infections
    #15672697 - 01/16/12 08:08 AM (12 years, 5 months ago)

Hi guys,

I just wanted to run this problem past you as I'm finding it difficult to pinpoint where I'm going wrong.

A bit of background - I've successfully inoculated and fruited using pf-tek method twice in the past with relatively little difficulties. Since then I moved house (keeping the same jars & vermiculite) and decided to give it another go.

Since then I've had 2 attempts at inoculating my jars using spore syringes from a reliable UK vendor. Both times every single jar has succumb to bacterial infection (evidenced by the horrible cheesy smell and lack of mycelial growth) despite following the sterilisation techniques to the letter as I did in my previous successful attempts.

Do you think it could be that the jars or vermiculite have managed to acquire a nasty bacteria that manages to persist through the sterilisation process here?

I've now bought fresh vermiculite, jars and brown rice flour and steam sterilised them all for 90 minutes . Is there anything else I should consider before taking the plunge and inoculating them?

Cheers,
Jay.

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OfflineSpawnThis
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Registered: 10/07/11
Posts: 33
Last seen: 6 years, 4 months
Re: Persistent bacterial infections [Re: cosmicjam]
    #15673177 - 01/16/12 10:37 AM (12 years, 5 months ago)

Make sure you are flame sterilizing the needle till it's red hot. Possibly a glovebox.  Sounds like you are sterilizing correctly.

A FOAF has has lost one FULL batch of jars due to the syringe being contaminated, never lost a full batch other than that... just a jar here and there.

Get/make a new syringe and try again.

Love.

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OfflineKizzle
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Registered: 08/30/11
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Last seen: 3 months, 17 days
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Re: Persistent bacterial infections [Re: SpawnThis]
    #15673613 - 01/16/12 01:15 PM (12 years, 5 months ago)

I'm thinking it could be syringe related too. It is possible to contaminate a syringe during use so getting it from a reliable source doesn't discount the possibility. (reliable sources can occasionally screw up as well)


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Offlinebaraka
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Registered: 07/15/00
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Re: Persistent bacterial infections [Re: cosmicjam]
    #15674995 - 01/16/12 06:40 PM (12 years, 5 months ago)

Might be the syringe, my be your sterilization process.

Next round do not inoculate one jar and see if it get bacteria like the rest


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This is the only time I really feel alive.

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Offlinemikesethnobotany
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Registered: 07/29/11
Posts: 1,267
Loc: Missouri, USA
Last seen: 2 years, 11 months
Re: Persistent bacterial infections [Re: baraka]
    #15676109 - 01/16/12 11:18 PM (12 years, 5 months ago)

Let me ask you this. How LONG are you sterilizing the vermiculite? I don't do these PF Teks or what have you, but if you must pressure cook the substrate (which I would highly suspect) then you 100% need to be doing it at the correct psi for a specific amount of time. Maybe you under-sterilized?

If the bacteria keeps appearing in the same spot as where the syringe inoculation occurs, then it is def. a syringe contam.

Solution:

Try sterilizing your substrate longer, and watch your jars to figure out whether or not the bacterial growth begins at the inoculation point.

As already stated be sure you flame sterilize to red hot , and absolutely don't wipe with alcohol afterwards.

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