contaminations - pink mould

[Guest electro]

the gnome down the road has asked me to post on his behalf ...

He has found that: of the 3 brown rice grain jars and the 5 wbs jars he innoculated - all 3 rice jars have become contaminated with pink mould within a week @ 16c and 3 days @ 27c (forgot to switch heat on lol .. no growth appeared until heat turned on)

The wbs is only just beginning to show signs of colonisation, so pink colours will not be detected in that for at least another 3 days.

He is wondering if the source of the infection is evident from the pictures (below) (he hopes it is the substrate coz if it was the spore syringe that would mean there are 5 more jars of pink mould growing [unless pink mould doesnt like eating wbs while myc does .. hrm ])

the rice was cooked for 25mins as per absorbtion method, loaded into 300ml dolmio jars, fitted with polyfil filters then pc'd for 1.5 hours at 12.6 psi.

The jars were innoculated in an ethanol soaked glovebox, injected through silicone sealed injection points in the lids (sealed prior to pc'ing) .. the silicone was wiped down with 95% ethanol and more silicone applied to seal the injection site.

The spore syringe should have been sterile, though this gnome did not make it so cannot be sure.

the wbs was soaked for 12 hours prior to pcing and was treated the same as the rice from there on in.

also how "dangerous" to health is this contam ? i mean some produce deadly / carcinogenic componds, but i couldnt find much on this pink one ...

Anyhow, comments greatly appreciated.

[ 14. June 2004, 14:45: Message edited by: electro ]

[mescalito]

From the shroomery..

Pink Mold; Red Bread Mold – Neurospora

Commonly to occasionally seen on agar and grain. Neurospora is fast growing, sometimes taking only 24 four hours to totally colonize a media filled petri dish. It is ubiquitous in nature, occurring on dung, in soils and on decaying plant matter. Since this fungus grows through cotton stoppers or filter discs, a single contaminated jar, though sealed, can spread spores to adjacent spawn jars within the laboratory. This condition is more likely if the filter discs or cotton plugs are the least bit damp; or if the external humidity is high. Furthermore, Neurospora spores germinate more readily at elevated temperatures. The pink mold seen in mushroom culture is most frequently Neurospora sitophila, a pernicious contaminant that is difficult to eliminate. All infected cultures should be removed as soon as possible from the laboratory and destroyed. A thorough cleaning of the laboratory is absolutely necessary. If contamination persists, remove all spawn and start anew.

[Rev]

bummer

[Guest electro]

lol it is a bummer.. but where is it from ?

is this some nasty spores that lived through an hour and a half of pressur cooking ? or is this a dirty syringe ?

[mescalito]

Could it be wicked in through the polyfill as the temperature/pressure in the jars decreases?

[ 14. June 2004, 22:37: Message edited by: mescalito ]

[Guest electro]

mescalito:

Could it be wicked in through the polyfill as the temperature/pressure in the jars decreases?

probably not .. the pink is appearing from the bottom up, and the jars were cooled in the incubator, being sprayed with 60% ethanol at 1/2 hourly intervals. The polyfil filters were also wet down with 95% ethanol at each of the spraying intervals .. lol even the incubator should have been sterile with the amount of ethanol being sprayed & dabbed in it

(almost a whole can of glen 20 was sprayed over 2 hours in a 30cm by 20 cm by 50 cm box & 200ml of denatured ethanol was used in the soakings ...)

The pink jars have been left to colnise outside the incubator, but ontop of excess heat wire (to maintain a decent temp) ... the pink has not grown substantially but there is more white myc . no idea if the white myc is the pink moulds mycellium without coloured spores (similar to the green mould that6 only gets its colour from spores ??)))..

will leave it to colonise n see

so far there is no pink in the wbs jars which leads me to believe that pink mould lives in the wholgrain brown rice from coles and is not killed by pcing (ive seen it before with rice cooks of only 30 mins and 45 mins pcing and almost always with just steaming)

[Rev]

im holding back final judgemnet

but 1 1/2 hours should kill all in jars that size especially with the pre-cook

Im leaning towards the syringe but well have to wait n see

[spiders]

ive had that one - thought it was mycelium piss at first - i thought wrong upon smelling it...

[Guest electro]

there is definatley more nasties in there now ..

i Really would like to know how all this crap got in those nioce sterile jars *scratches head*

lol there is some bacillus aswell as the pink stuff. ..

the rice jars were emptied out, the crap discarded & the remaining good (nice smelling) myc broken up & cased with verm ... (why not hey .. nothing to lose lol)

wbs jars still seem okish .... myc creeping between most grains now ... some small white patches ... should know soon if these are free from pink or not (they are beginning to show signs of bacillus now with some grains touching glass getting very wet .. if they go white then i wont be happy lol... hopefully it is just condensation settling on those grains ... either way though, the myc seems to have taken a decent hold so the bacillus shouldnt spread too much)

thanks all for the helpful replies, they are appreciated.

also bm .. mycellium piss ?

or larger version ...

http://www.shaman-australis.com/publicpics/wbs.JPG

[ 15. June 2004, 21:49: Message edited by: electro ]

[Rev]

Mycelium piss

coloured exudate from overaged or challenged mycelium.Sometimes foul smelling (and tasting - i shouldnt i know!!) but normally innocuous and completely sterile, usually coloured but transparent. cloudy piss in mushrooms as in humans is not welcome and often indicates an infection, in this case yeast or bacteria. Ganoderma often has Fanta Orange, shiitake brown, oysters yellow.

Very likely to have intereting antimicrobial/ medicinal properties in some species

[mescalito]

Gnome entry #702.

I've had a couple of goes at wbs and don't really like the multicolour spectacle much!

The best results for me so far have been with brown rice as thus:

per pint jar-

1/3pt brown rice was run through a food processor until the husks are noticably removed and the resulting flour was winowed and discarded.

1/3pt vermiculite.

1/4pt brf.

mixed together with enough water to 'just' wet.

loaded in 1pt jars with a foil/verm cap under lid and 4 injection holes poked through both..foil over top.

PC'd 15psi for 2hrs with 2 HOT water refill intervals.

holes taped and 1pt jars put away for 1 week in a warm semi-dark shelf for observation.

innoculated in glovebox and into incubator @25'c 3 weeks.

colonised cake crumbled and cased with 50/50 verm/coir.

I've had zero contams through to fruit so far using this method which is interesting considering the grain wasn't soaked at all :confused:

[ 16. June 2004, 03:38: Message edited by: mescalito ]

[Rev]

You dont need to soak rice

its pretty clean - such that id be suprised if the rice was the problem

Electro are you sure your PC is running at pressure? any way to tell?

The soaking merely encourages dormant bacilli to germinate thus making them easily killed by heat

If moulds are your problem then i dont know if soaking will help, only longer sterilisation times

I have had mould spores make it through polyfill occasioanlly but its usually obvious - like blue green mould and always starting at the top, and when i innoculate polyfill jars i do it on the kitchen benchtop with nothing but a spirit lamp to flame the ends - no glovebox, HEPA etc and very rarely does the polyfill let me down

Anything on the bottom is usually indicative of improper sterilisation or innoculant ferried contam

if the WBS do not contam then id support the notion that the PC wasnt operating at enough pressure. Time was long enough even at 12psi.

[Guest electro]

Electro are you sure your PC is running at pressure?

---no idea .. the heat gets turned full ball until aabout 10 mins after boil, then turn the heat bback to the lowest setting, so it keeps just letting steam out.. if it stops hissing it gets the heat turned up a bit.

any way to tell?

--the metal sitting thing stops boucning about & hissing ... its one of those cheap $90 weighted ones ..

if the WBS do not contam then id support the notion that the PC wasnt operating at enough pressure.

---shouldnt the wbs be more prone to contam if there wasnt enough pressure >? (also cooking was done in 2 lots ... 4x wbs, then 3xrice + 1X wbs) so if it was a lack of pressure issue the single wbs jar from the rice cook should also be very contamd ...

[mescalito]

Whatever pressure the weight exerts due to gravity is the pressure you're getting out to cause steam release.ie.newtons first law of physics..so is there an easy way to calculate the psi/kpa pressure it would take to lift 'X'grams weight

[Guest electro]

re "so is there an easy way to calculate the psi/kpa pressure it would take to lift 'X'grams weight"

im sure there is as the weight itself is what has the pressure rating on it .. measured in kpa... 80 kpa which i thought (though I could have been very wrong) was around 12psi

i wish it was the pc .. it would explain SO much ... except why stuff non innoculated jars last weeks w/out contam).

Does anyone agree that this here is bacillus ? Or is it just condensation & me being paranoid ...

if it is bacillus the last 5 grain jars all went today will leave in the incubator for another week and see later if the whole lot is just a pool of goo or if the myc has fought it off ...

also, if it is bacillus that means that

8/8 jars all contamed with bacillus

3/3 rice jars contamed with bacillus and pink mould

please place your bets on the source so i can stop wasting money on irrelevant stuff .. thanks all ....

overview of procedure

cook rice 25 mins, drain for 2hrs, load into jars polyfil lids added & injection point siliconed

soak wbs for 8hrs, drain for 2 hrs, load into jars, polyfil lids added & injection point siliconed

2 pressure cooks, 1.5hrs each with 4 X 300 ml jars in each.

Cooker has 80kpa weight, which hissed for the whole time.

jars transferred while hot to incubator & swabbed with alcohol every 1/2 hr paying close attention to the polyfil filters.

when cool jars tranferred to alcohol & bleach swabbed glove box & injected once each through ethanol wiped silicone injection point. needle reflamed inbetween each injection.

jars reswabbed with ethanol & transferred to incubator.

[ 16. June 2004, 21:42: Message edited by: electro ]

[Rev]

? you have poyfill lids but inject through silicone port?

With mine i inject through the polyfill - easier i think

re rice

I cook using absorbtion method. 1 cup rice to 1 3/4 caupswater then 1:2 ratio for every extra cup. Add a decent pinch of lime or gypsum. Bring to boil them reduce temp right down and leave covered 20 mins till all water absorbed. works well especially with a rice cooker:)

After cooking i let pressure equalise then take ot the fars and put on the bnch to cool and do the next lot. Do you cap jars with alfoil? I do

My jars work fine this lazy way so i doubt your technique with greater care is the downfall

I also dont innoculate in the glovebox, or wipe them down after

you are right about teh rice wbs contam thing - i missed that detail

hmmm bad luck?

Bacillus? i think thats early to tell

it may just be the extra moisture from the syringe added to condensation. Youll know if its bacillus when the growth stalls the grain darkens and the ultimate teller - the fruity turning sour smell. So far growth looks healthy and active. youll know soon

I dont trust my eyes as much as i trust my nose

i sniff all my filter patches and polyfills regularly and this measure weighs heavy on my diagnostic scale

Another thing that has often run me to ruin has been incubation at high temps as it seem sto wake up any remaining dormant spores. Try incubating at a stable 23 - 25 it might be better even though its not the optimum temp

Oh another thing - my biggest bugbear of all time ever since i started has been and continues to be - Dirty stinking Trichoderma on bulk susbtrates

If not for that it would be a breeze

The species i grow now are not as troubled by it like oysters as they are pretty fast and faily immune to its invasion but other species are really susceptible to it

[ 17. June 2004, 00:36: Message edited by: reville ]

[mescalito]

Electro-yes 80kPa is 12psi.

My gnomes best jobs so far have been through the method I mentioned above^ basically he thought they were WAY too dry but inocculated them anyway and away they went.

He has a 1 pint jar atm that's 98% colonised after 4 weeks (it is a pint jar) @ 20-25'C

He tends to err on the dry side now

[Guest electro]

re mescalito:

"My gnomes best jobs so far have been through the method I mentioned above^ basically he thought they were WAY too dry but inocculated them anyway and away they went." --- yea thats how the wbs was .. it looked drty, but the heat of the incubator forced more water out

re rev:

"you have poyfill lids but inject through silicone port?" -

-- yes, didnt want to take ANY chances disturbing polyfil to let contams through it ...

re rice - yea thats how this lot got cooked (apart from the pressure equalising step)

re "Bacillus? i think thats early to tell

it may just be the extra moisture from the syringe added to condensation." -

- thankyou SO much for saying this .. these would normally have been thrown that out for fear of growing more bacteria that could spread .. not wanting nto contam everything it would have been huled into the bin with a whole bunch of bleach .... you were right .. this WAS just condensation .. i wonder how many good jars have been thrown because of this id mistake LOL ..

thanks all for your imput, advice and help .. thankyou all so much

Below, happy wbs ( i think)

and the straight verm casing of the non contam'd rice (the "it couldnt hurt" idea was a good one)

.. i still have no idea how the rice got so badly contamed though ...

Happy WBS

vermiculite casing (1/2 cm verm ontop, 1cm verm below, 3cm substrate)

[ 20. June 2004, 16:16: Message edited by: electro ]

[Auntyjack]

a person once told me he found this bright pink contam on his casing and after a few flushes, didn't seem to be too agressive tho and things continued on around it as normal . Anyone else run into this problem?

AJ

[mec]

lol it is a bummer.. but where is it from ?

I believe the pink mould comes from inside the grain, thats my bet.

[Auntyjack]

weird thing is the pink mold was eventually overtaken/overlayed with a fluffy dense white mold, I don't think it was cobweb or the myc??..hmm wish I had taken photo's sorry guys.

AJ

EDIT: also mec if the punk mold comes from inside the grain that would suggest that PC'ing does not sterilse totally, and why did it not show until the last flushes? Surley the other contams that hit at this time is a factor of exposure and dunking?

Edited July 14, 2007 by Auntyjack

[soma_seeker]

i've battled with lipstick mould quite a bit. my initial grow attempts were innoc'd with LC and EVERY single jar became contam'd with it every time. I left one sterile jar aside and it didn't contam so it was obviously from innoculation

I then moved to multi-spore (and innoc'd in the exact same way) and did not see it again.

Lipstick mold can be easily overtaken by myc if it has already somewhat colonised the jar. i'm deff not endorsing playing around with contams considering the risks involved - however i have on numerous occasions succesfully fruited jars that were contam'd with lipstick in their early stages of growth.

it is a mold and as such (opposite to bacteria contams) is introduced at innoc and not due to incomplete sterilization.

i'd work on your syringe prep and innoc techniques.

(sorry if i've repeated anything previously mentioned, i only quickly scanned thru past posts)

peace

[Auntyjack]

hi soma,

yeah this is the strange thing, it didn't appear until about the 5 flush in the casing....well past the innoc stage. I think that friend is purty finniky with syrnges and spawn so has only lost one or two jars to contams (mainly too wet to start with).

AJ

[soma_seeker]

ah, my bad - guess i should have spent a bit more time reading

i was told that lipstick mold spores are often present in ould wooden cupboards and the like, maybe since ur casings had been around so long they picked some up from the surrounding environment.

5 flushes on a casing, dam

[Auntyjack]

friend said yeah they just kept comming and comming, he didn't even recase.

I don't think winter will be as kind tho...

AJ

Edit:spelling

Edited July 15, 2007 by Auntyjack