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OfflineYSJ
ThoughtShopper

Registered: 01/31/03
Posts: 57
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Last seen: 2 years, 3 months
A contam question which I cannot find addressed anywhere....
    #1265178 - 01/31/03 10:52 AM (13 years, 10 months ago)

To Whom It May Concern,

First, I'd like to offer thanks to those who have made this place possible - it's content-rich, nicely designed, and both user-friendly and reasonably organized (N.B.:  No, not trying to kiss any ass -- my personal arrogance forbids it, sorry :wink:  but having designed/developed/maintained enough sites professionally in my life, I know the sort of work which can go into such things, and simply wanted to express an honest note of appreciation.

Oh, and lest there be any doubt:  none of the text cited below has anything to do with me; it all actually pertains to Someone Who Isn't Me (or, shall we say, 'SWIM', for short?? :wink:

So here's my contam question.  I really hope someone can give me a positive answer, as I cannot find ANY definite answers or identifying images, and that includes scouring your FAQs, your site, doing loads of W3 searching, as well as scouring archives of appropriate areas of the alt.* hierarchy.

Short version:  I know it is possible to have contams after-birthing of a cake; that granted, are there any which A) only show up on the inside of the stipe, and show up nowhere else, and B) do not appear to inhibit fruiting of said cake?

Long version:  (more info).

I know it is possible (if less likely) for a fully mycelium colonized cake to succumb/be infected by either  competing/antagonistic fungii or bacilli, via any number of vectors; however, what I cannot seem to understand (or find any identifying pictures of, or information regarding) is something which has occurred in one cake (further details about prep below).

While a mycological neophyte, I'm reasonably adept with inorganic & organic chem,  as well as proper lab procedures, and the like. 

I'm torn between trying to keep this very brief, as it is my first post, and my desire to give as much detailed info as possible, in the hopes of someone being able to positively ID/diagnose/help out with this.  I suppose those who are uninterested (or if the Mods decide to eliminate this post as being too OT), then so be it, and I'll err on the side of the latter in the hopes that someone with some crucial bit of knowledge will read it.

Suffice to say, SWIM managed to do the following to near-textbook-perfection:

- substrate prep w/proper ratios et al;
- substrate/utensil/workspace sterilization (done _WAY_ to the point of overkill - while I don't have a $10,00,000 clean room a la IBM's lithography labs, I possess both the skills and the resources to make what amounts to the best possible clean room one could possibly make in a non-corporate setting);
- proper substrate inoculation procedure without (AFAIK) introduction of any contams whatsoever;

?To cut to the chase, SWIM found himself with the happily achieved end-result of being in possession of 12 successfully colonized half-pint jars (tapered as appropriate, natch), and with the judicious use of some thermolyne heat tape, a rheostat, and a couple of lab-grade thermometers, watched as all but two of the twelve successfully colonized in record time at ~83 deg. F.  (Of the two which never fully colonized, I did a post-mortem, and it was presumably due to poor moisture ratio - the substrate was too dry, so that part needs work, obviously :wink:

The other ten became white as driven snow within an appropriate amount of time, and, with no uncolonized spots left for contams to take root in, and with primordia appearing, were incrementally birthed not long ago.  FYI, these were inoculated/colonized with B+.

Anyhoo, SWIM having some experience designing digitally controlled, high-precision controlled-climate environments before, SWIM designed his own fruiting setup, keeping the rH between 95% and 99%, along with mist shielding/direct air-flow-baffling, etc., with cleaned, prehumidified 02 being gently & periodically forced in, the CO2 having its own exit near the bottom along with an excess water drain.

In any case, it appears that SWIM's setup is at least ideal enough to have initiated plentiful fruiting, all going well in that regard, but for that one mentioned before?..

While *NO* contams were witnessed during colonization, and every precaution was taken (both before/during/after all stages, and even just doing maint. work on the terrarium setup occasions SOP sterilization procedures - doing it by the book & all that :wink: -- after the initial flush & timely removal of same, SWIM found his puzzler:

Inside of the stipes of a set of fruits (all from the same cake) was a bright green filling, and it appears the entire stem is filled with it.  SWIM's initial thought was "Uh?.Bruising should have been more blueish, and on the OUTSIDE, not the inside, so this *must* be a contam?"

SWIM having read horror stories (one in particular from A.D.P. comes to mind) of someone who ate blackened, obviously rotten fruits with the inevitable result of violent illness, SWIM will naturally stick by the "if in doubt" methodology. 

While isolation of the offensive party has already occurred, and SWIM is fine just throwing it out, SWIM is really curious about this brilliant green found *ONLY* inside of the stems, and the mycelium where SWIM cut it (with a scalpel, after harvesting).  The cake has not yet been dissected, on the offchance someone might reply that this is normal for B+, which SWIM has had no prior experience with.

Further info for those interested:  upon exposure to oxygen, the light green quickly faded to dull green.  Again, having re-read your FAQ on contams, SWIM can honestly say that this doesn't appear to be Trichoderma spp. Or the like, nor was anything like that witnessed during colonization (which went flawlessly).  While all other contams that remotely fit this description are all notably surface-dwellers/infectors first, I'm currently doing further research on Pseudomonas aeruginosa, though I think I may be grasping at straws here (or, being such a neophyte, missing something incredibly obvious to everyone else :wink:

I welcome any replies to this, and SWIM would naturally be eternally grateful to anyone helping to solve this mystery, and offers advance thanks to those who have read this far.

Please either reply here and/or email me ( ysj -= at-symbol =- indesp.com )

Regards,


~YSJ~
--
post scriptum:  I'll shortly break out the Digital Cam & take some pix, in the vain hopes that might help identification.


--------------------
"There are no angels unless they are Angels of Death...And I would never again doubt my place among them or lose my resolve to serve in their wild ranks."         
                                                            -T. Ligotti


Edited by YSJ (01/31/03 10:56 AM)


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InvisibleChibboleth
member
Registered: 10/14/02
Posts: 232
Re: A contam question which I cannot find addressed anywhere [Re: YSJ]
    #1265947 - 01/31/03 03:12 PM (13 years, 10 months ago)

That's extremely strange, and odd green filling is certainly not normal for B+. I would guess it to be a mold of some kind. What is the texture like?

I'd dissect the cake and post pictures.


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Offlinebowling-name
sleuth

Registered: 09/24/02
Posts: 282
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Re: A contam question which I cannot find addressed anywhere.... [Re: YSJ]
    #1266283 - 01/31/03 06:30 PM (13 years, 10 months ago)

One possibility is to take a tissue culture from the infected stipe and grow it out on agar (assumming SWIY has the capacity/equipment/time) for this to see what the outcome will be for this potentially infected mycelium. This would yield interesting data about what this 'infection' might be if its full life cycle could be observed.

Mold growth seems unlikely as it is usually a direct competitor and not a parasite of fungal fruit bodies. My first thought would be that there was an environmental contaminant or VIRAL infection, but the strict and sterile controls of the experimenter and non-uniformity of contamination seem to preclude this conclusion. Quite mysterious. Post pics!


Edited by bowling-name (01/31/03 06:32 PM)


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OfflineYSJ
ThoughtShopper

Registered: 01/31/03
Posts: 57
Loc: Undisclosed at this time.
Last seen: 2 years, 3 months
Re: A contam question which I cannot find addressed anywhere [Re: bowling-name]
    #1271902 - 02/02/03 07:24 PM (13 years, 10 months ago)

>One possibility is to take a tissue culture from the infected stipe and grow it out on agar
>(assumming SWIY has the capacity/equipment/time)

Funny you'd mention that?.SWIM was actually thinking the same thing, but for the time/concentration factor?.Time?.Aye, there's the rub:  SWIM is currently facing down some major DB development, as well as some serious Apache module development (mostly mod_perl stuff) for a production site, so time is the thing I've got the least of :frown:

>will be for this potentially infected mycelium. This would yield interesting data about
>what this 'infection' might be if its full life cycle could be observed.

Totally.  However, whilst scouring the archives on unrelated topics, I came across the following post, which gives me pause (be sure and read the followup threads & tell me what you think:

http://www.shroomery.org/forums/showthreaded.php?Cat=&Board=Forum2&Number=14306&Forum=All_Forums&Words=mildew&Match=Entire%20Phrase&Searchpage=0&Limit=50&Old=allposts&Main=14306&Search=true#Post14306

In the event that link gets fux0red somehow, the gist of the post(s) are as follows:

"So two of the [B+] cakes have finally fruited, and I harvested 3 shrooms when the veils separated.
Their stalks, if viewed in cross-section, are white on the outside, green on the inside, and hollow in the very middle. Is the green coloration normal?"

Reply from Mycelium5150:  "totally normal for b+, there hollow and the color is from you cutting them, they turn before you can even see it. there fine, eat away."

Just offering that bit for whatever it may be worth

>Mold growth seems unlikely as it is usually a direct competitor
>and not a parasite of fungal fruit bodies.

My thoughts exactly?So, thank you for letting me know my reality check hasn't completely bounced :wink:

From SWIM's (admittedly limited) understanding, he figured that (to grossly oversimplify, and not counting *all* contams, obviously), if the mycelium gets "sick", it usually diminishes fruiting, or shows other signs of illness, low yield, etc.

>My first thought would be that there was an environmental contaminant
>or VIRAL infection,

My fear as well.

>but the strict and sterile controls of the experimenter and non-uniformity of
>contamination seem to preclude this conclusion.

Of SWIM's many qualities, lab sloppiness isn't one of them (being *really* anal when it comes to things like that, believe me), which is why SWIM was baffled, just from what little SWIM knows of mycology.

>Quite mysterious. Post pics!

SWIM definitely will be doing so, but it won't be for another few days, just due to time concerns.

And SWIM is kicking himself now for not having taken pix the moment he harvested those in question?.I think that would have been the most helpful, probably.  Next time, he'll tripod the Nikon and have it ready to go, just in case.

Again, thank you for your reply - I honestly appreciate it - and as I can, I'll post more info/pictures in the hopes that others may benefit.



~YSJ~
 


--------------------
"There are no angels unless they are Angels of Death...And I would never again doubt my place among them or lose my resolve to serve in their wild ranks."         
                                                            -T. Ligotti


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OfflineYSJ
ThoughtShopper

Registered: 01/31/03
Posts: 57
Loc: Undisclosed at this time.
Last seen: 2 years, 3 months
Re: A contam question which I cannot find addressed anywhere [Re: Chibboleth]
    #1271918 - 02/02/03 07:35 PM (13 years, 10 months ago)

>That's extremely strange, and odd green filling is certainly not normal for B+.

That was what SWIM was thinking, as well; however, in a somewhat unrelated search through the site, SWIM did come across a post from a couple of years ago:

http://www.shroomery.org/forums/showthreaded.php?Cat=&Board=Forum2&Number=14306&Forum=All_Forums&Words=mildew&Match=Entire%20Phrase&Searchpage=0&Limit=50&Old=allposts&Main=14306&Search=true#Post14306

The gist of the post(s) are as follows:

"So two of the [B+] cakes have finally fruited, and I harvested 3 shrooms when the veils separated.
Their stalks, if viewed in cross-section, are white on the outside, green on the inside, and hollow in the very middle. Is the green coloration normal?"

Reply from Mycelium5150:  "totally normal for b+, there hollow and the color is from you cutting them, they turn before you can even see it. there fine, eat away."

Just offering that bit for whatever it may be worth.  SWIM honestly DID scour the search engines before, but had been unable to find that one; so, huh.  Beats hell outta me what to think on this one without dedicating some serious research, which I'm DYING to do, to be honest. . . But then there comes that damned RL bullshit, what with work & all ;(

>I would guess it to be a mold of some kind. What is the texture like?

Running water test = negative.  No flaking off, no rubbing off, no nothing.  Just as if it were a bruise, but not blue. 

[sung to the tune of "I think I'm turning Japanese"]:

~/ I think I'm going colorblind /~

:wink:

> I'd dissect the cake and post pictures.

Yeah, that's a definite.  SWIM will do so, the moment he digs himself out of a MOUNTAIN of work looming this week for a production level site - total nightmare deadlines.

More info as I have time to post it. 

Oh, and BTW:  thank you very much for your reply to my posting - I honestly do appreciate it :smile:


~YSJ~
 


--------------------
"There are no angels unless they are Angels of Death...And I would never again doubt my place among them or lose my resolve to serve in their wild ranks."         
                                                            -T. Ligotti


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Invisiblezeta
Stranger

Registered: 05/25/02
Posts: 3,972
Re: A contam question which I cannot find addressed anywhere [Re: YSJ]
    #1272244 - 02/02/03 10:28 PM (13 years, 10 months ago)

You seem like a very valuable addition to the community. Welcome!  :smile:
I am puzzled about this: natch?


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OfflineMAIA
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Re: A contam question which I cannot find addressed anywhere [Re: zeta]
    #1272407 - 02/03/03 12:37 AM (13 years, 10 months ago)

Yep, welcome !
It also puzzles me....
Did the green had any sporulation whatsoever. Grab a wet paper and rub that area, you'll see then.

MAIA


--------------------
Spiritual being, living a human experience ... The Shroomery Mandala



Use, do not abuse; neither abstinence nor excess ever renders man happy.
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OfflineYSJ
ThoughtShopper

Registered: 01/31/03
Posts: 57
Loc: Undisclosed at this time.
Last seen: 2 years, 3 months
Re: A contam question which I cannot find addressed anywhere [Re: MAIA]
    #1272507 - 02/03/03 02:55 AM (13 years, 10 months ago)

>Yep, welcome !

Well, thank you :smile:

>It also puzzles me...
>Did the green had any sporulation whatsoever. Grab a wet paper and rub that area, you'll see then.

Well, I did try the running water test, with negative results.  I should have added that yes, I also did the rub - again, negative.  No flaking off, no rubbing off, no nothing.  Just as if it were a bruise, but not blue (perhaps I've gone colorblind? 

More news as I have time to make it available.


~YSJ~ 


--------------------
"There are no angels unless they are Angels of Death...And I would never again doubt my place among them or lose my resolve to serve in their wild ranks."         
                                                            -T. Ligotti


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OfflineYSJ
ThoughtShopper

Registered: 01/31/03
Posts: 57
Loc: Undisclosed at this time.
Last seen: 2 years, 3 months
Re: A contam question which I cannot find addressed anywhere [Re: zeta]
    #1272514 - 02/03/03 03:07 AM (13 years, 10 months ago)

>You seem like a very valuable addition to the community. Welcome!

Gosh!  If I have to say thanks for the many warm welcomes I've received, I'm going to start sounding like a kiss-ass ;D

But seriously, thank you :smile:

I honestly hope I can be of service to this community.  I've been lurking a bit, and like everything I see so far (the grow logs are a *great* idea, btw).

>I am puzzled about this: natch?

I'm sorry, perhaps I'm misreading the above, but do you mean you're puzzled by the use of that word, or puzzled by the whole green-inside-stipe issue?  If the latter, well, I'm working on it?.if the former, I was just using slang for 'naturally'.

Well, work is already starting to kick my ass, so I have to get up on?.

Again, thanks for your kind words, and taking the time to reply.



~YSJ~
 


--------------------
"There are no angels unless they are Angels of Death...And I would never again doubt my place among them or lose my resolve to serve in their wild ranks."         
                                                            -T. Ligotti


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OfflineBigJohnson
Whoa! You guysneed to lightenup!

Registered: 02/04/03
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Re: A contam question which I cannot find addressed anywhere [Re: YSJ]
    #1290259 - 02/08/03 06:45 AM (13 years, 9 months ago)

Perhaps a population of algae is living symbiotically with your fungus. It makes sense.


--------------------
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http://www.lp.org/issues/relegalize.html


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