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wbs peroxiode and no heat

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Guest electro

a mate of a mate is conducting an experiment with a dung lover

wbs has been soaked in water (water filled to 1 inch above the seed level in the soaking pot).

To this was added 200 ml 3% peroxide, and the seed left overnight in a pot.

The hope is that all iving things within will be oxidised by the h202, leaving a sterile substrate to innoculate.

After much bubbling and swelling of the seeds excess water was drained off and the seeds scooped with sterile spoons into plastic bags.

Filters were added to the bags for air exchange and the bags were innoculated with a spore syringe each.

no results yet, but he says he'll get back to us.

[ 30. March 2005, 14:07: Message edited by: electro ]

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Guest electro

updates:

seeds are sprouting .. oops

mycelium appears to be forming in some spots

also one bag had some mycellium transferred from an outdoor straw and dung mushie patch to compare with bag innoculated with spores

the best news, as yet no infections... slight smell but most likely due to sprouts beginning to decompose.

[ 06. November 2003, 20:59: Message edited by: electro ]

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How is it that you were able to innoculate h202 seed with spores? Doesn't h202 prevent spore germination?

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Guest electro

coz the h2o2 was left to chew on the seeds overnight...

by the time of innoculation there "shouldnt" have been any peroxide left .. it should have donated it's extra oxygen to everything it touched ( seed husks existing spores, bacteria etc)

[ 30. March 2005, 14:09: Message edited by: electro ]

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ahh hence the bubbling. Hopefully there were no established moulds on the WBS?. Nice idea, good for PC'less folk.

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sorry, i'm a little slow, what is 'wbs'? bird seed?

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this will be interesting

have you read up about the peroxide tek?

namely what peroxide does and does not do?

after this expt i strongly recommend you divert research to the '10 minute' spawn concept

using highly processed materials as i think these have much more potential for this kind of researchg and there are so many

grain can be hard to sterilise at the best of times using a pc

peroxide is not a sterilant

peroxide will be most useful when dealing with wood decaying fungi and for them peroxide wood based spawns are great. wood decayers seem to have better tolerance to peroxide than compost and dung dwellers - of course there re the indestructible exceptions

the funky smells due to arise will be peroxide tolerant aerobic bacteria making a lovely WBS soup for you :)

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Guest electro

-->have you read up about the peroxide tek?

namely what peroxide does and does not do?

no he hasnt

--> the funky smells due to arise will be peroxide tolerant aerobic bacteria making a lovely WBS soup for you :)

yup, he has noticed them (they got very bad at one stage). Some vermiculite was added to soak up excess moisture to help prevent rotting & bacterial breeding. this did help and the smells are also diminishing as the mycellium takes over though (3/4 colonised now - no competing fungus seems to be aparant though - which is amazing as green and cobweb mould even grows happily in pasturised cow dung in my mates neighbourhood).

he has addressed this (also experimentally) by testing clearasil's antibacterial claims. The main ingredient besides water is sodium laural sulphate which theoreticly kills bacteria on conact and is not "too much" of an antifungal agent. He is expecting mycelial growth to come almost to a hault then slowly recover / recolonise and for the smell to hopefully dissapear.

To one bag was added 1 ml clearasil complete deep clensing daily facewash with microbead formula diluted in 90 ml H2O.

To another slightly less colonised bag was added 10 ml of clearasil diluted in 150 ml of water.

After thorough mixing the clearasil additions were followed by 300 ml and 600 ml (by volume) additions of vermiculite to "soak" up the excess moisture.

notes if this is successful the addition of clearasil will be peformed prior to innoculation or 3 days after (depending on how fungacidal it turns out to be).

if this step fails it will be repeated with 20 ml cream, failing this either a KMNO4 + dH2O (aqueous condies crystal) solition to wipe out everything bar some mycellium or hitting it with metho in hope that the ethanol will kill the bacteria and hopefullly spare some of the strongest mycellium .... perhapse after the soap and before the metho and oxidisers some antibacterial spray (glen 20 ?) might do the trick a tad more selectively...

As for growth it is been phenomenal .. 2 kilos (dry) of wbs almost completley colonised in under 2.5 weeks .. estimated 3.5 week complete colonisation at 22.5 C with 4 * 1/2 hr fanning + us humidification.

conclusion thus far ...

very interesting project, needs modification though to stop bacteria .. possibly antibacterials, kmno4 or ethanol to begin with or (shudder) perhapse some heat treatment, hopefully the antibacterial (to be added shortly) doesnt kill mycellium

also rev, he will focus his attn on that tek next, once refining this into a do able tek... wbs was specificly chosen for being so difficult to sterilise, if this tek can address that (ie the HUGE amts of peroxide and the possible introdution of an antibacterial once smells are noticed (or prior) then all will be good and well and plentiful.

[ 30. March 2005, 14:11: Message edited by: electro ]

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Guest electro

my mate has given up :/

trying to kill an bacterial infection is much more dangerous to ones health than preventing it (even if the battle is won, who knows what fruits will carry with them of the antibacterials)

my mate is reverting to pf for a while..

The pf will however not be without experimentation, 6 jars will be innoculated and boiled (steamed for 1 hr). the last 3 to be boiled will have had peroxide added to the mixture (10 ml in total). innoculation will follow 24 hrs later when the jars have cooled.

the three will be compared in growth speed and in purity (peroxide %contams vs plain pf)

The current (grain) project will be grown until maturity and added to pre prepared straw & cow dung (substrate that was intended for the dry and slow colonising sock tek).

Finally if all 6 jars colonise properly an experiment will be done with one jar, adding it to 4 litres of cardboard (some refs indicate that cub can grow on cardboard but not much info boiled(100 c 10-20 mins) in a pot, drained and will have peroxide then added.

[ 30. March 2005, 14:17: Message edited by: electro ]

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trying to kill an bacterial infection is much more dangerous to ones health than preventing it

definitely - i have also come upon some seriously funky things that no doubt do not naturally occur in concentrations as big as they do when cultivation goes wrong - be careful there

The pf will however not be without experimentation, 6 jars will be innoculated and boiled (steamed for 1 hr).

Fractional sterilastaion is a technique that needs refining and expanding on

particularly

- a reproducible protocol for larger jars (> half pint)

- Microwave oven fractional sterilsation

the last 3 to be boiled will have had peroxide added to the mixture (10 ml in total).

is this so the freed oxygen plus heat plus pressure gives 3 way killing power? ive often wondered whether adding peroxide before sterilsatiuon to enhance this process was viable

innoculation will follow 24 hrs later when the jars have cooled.

the three will be compared in growth speed and in purity (peroxide %contams vs plain pf)

The current (grain) project will be grown until maturity and added to pre prepared straw & cow dung (substrate that was intended for the dry and slow colonising sock tek).

if you get any decnt growth on grain at all ive seen it best handled by moving to a dry and well aerated mix

It seem sthe way in which bacteria dmage mycelium is by changing the pH and souring of the media by metabolic wastes

however it is the confined and poorly aerated containers that actually help finish off the mycleiums resistance

If you put grain onto a selective (poor) pasteurised substrate with good aeration and abrorptive capacity the pendulum quickly swings in favour of the mushroom. Volatile metabolites disappear or are broken down by resident aerobes and the mycelium often just eats the remaining bacteria as it grows over them

amazingly ive seen the same with moulds

mould s grow quick ;ly and ibve seen bags and plates where i though the mushroom lost. however after the moulds cover the plate the myclijm adajusts from being a sparophyte to being a predator and eats the moulds. Agaricus are classic like this - very slow on agar unless teh agar become s contaminates then they take off.

Enoki and stropharia rugoso did the same

it seems theres 2 ways to get to the top of the compost heap

If in doubt throw it out(side)

Finally if all 6 jars colonise properly an experiment will be done with one jar, adding it to 4 litres of cardboard (some refs indicate that cub can grow on cardboard but not much info boiled(100 c 10-20 mins) in a pot, drained and will have peroxide then added.

b plus does ok on paper and cardboard but youre courting with danger as even in the best situation trichoderma is ALWAYS better adapted and will get there sooner or later

rev.. sawdust isnt a viable option for the gnome down the road.

I find the paper pellets that they make for kitty litter to be a staple in spawn production- look in the pet food aisle. Blend perfectly with the peroxide method and are a great option for woodlover spawn. The urban mans sawdust

Have you tried the fermnetation/conditioning tek?

i didnt make it up but ive been playing with it while it seems neglected by others

using straw, bagasse, soybean mulch, chaff , dung etc - whatever you need for your saprophyte

soak ingredients overnight to ghet water content up, drain for a few hours

miz well and sprinkle in a handful of gypsum or lime

pack into a sealable clear tub and place with lid on tight in a spot receiving full sun all day

the temp will rise quickly

leave for 3 days

it seems most fermentable sugars are gone by then and the mix packed with benign bacteria

So now the media is selective and there is little room for competitors to re-establish

as the mycelial netyworks colonise the mix the fermented smell is replaced by the sweet mycelial smell.

very low incidence of trichoderma

suitas all straw species and may even be good for agaricus outdoors (mound culture)

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Guest electro

double post.. sorry ...

[ 15. November 2003, 14:32: Message edited by: electro ]

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Guest electro

rev: the 20 l bags of cardboard kittlylitter pellets is exactly what my mate was planing on using as cardboard, though hes not sure if cubs like cardboard (as they are dung dwellers, but reports suggest that they can eat damp hemp string so .... if they will eat it though its worth a shot - it's cheap, easily accessable and very absorbant (it will suck peroxide up well and possibly retain shape to keep the mix areated).

Updates:

WBS

The two bags were used to inoculate two cow dung/ straw patches mulched with tree fern & palm leaves.

Both were completley colonised (1 day shy of 3 weeks from spore injection IN PEROXIDE till colonised)

The bag treated with 1 ml clearasil did stink less the following day, but 1 day later stank just as bad as ever

The bag treated with more water and 10 ml clearasil had a slightly sweet (slightly infected) mushroomy smell.

Myco growth was only affected for 1 day, but shot back the next (from 80% colonised to 100% after thorough mixing-it seems the mycellium liked the bacteria much better dead than alive, more food maybe or less competition?)

NEW experiments

EXPERIMENT 1

6 jars were prepared pf for simple minds style (shroomery.org)

3 after steaming for 1 hr were injected while still warm with 6 ml (one ml in each of the 6 injection points) of 1 part 3% peroxide to 2 parts water.

The jars were left overnight to cool.

EXPERIMENT 2

3 liters of the pf mix was mixed up and placed into a glad snap lock bag. to this was added 10 ml 3% peroxide.

the snap lock bag was left open and placed into a $6 (coles) microwave vegetable steamer and microwaved for 60 mins, 10 to boil 50 of boiling.

A terrible smell filled the air but the source of the "burning smell" was not discovered until the steamer was opened.

The pf cake was dry as a rock and the plastic bag had shriveled up and melted away to nothing bar a shiny coating on parts of the cake ...

Total Failure: lucky not all the verm has been used up

EXPERIMENT 2.5

1.5 liters of the pf mix was mixed up and placed into a glad snap lock bag.

the snap lock bag was left open and placed into a $6 (coles) microwave vegetable steamer and microwaved for 20 mins (10 mins to get to boil, 10 mins of boil).

The bag suffered but was still air tight.

WHile still boiling hot the bag was placed inside a larger glad snaplock bag (which had been soaked in 3% peroxide [ie still wet with the stuff on the inside]).

The top of the iner bag was left open for air exchange between it and the outer bag (which will be opened for 2 secs daily).

The 1.5 L pf cake was injected with 20 ml of a 1:2 3% peroxide:water solution in approx 20 points.

The bag was left to cool overnight.

EXPERIMENT 3

In the morning the microwave steaming dish (which fits inside the steamer) was lined with baking paper.

1.5 L of long grain rice was placed into this and covered liberally with water.

The water formed a pool as it soaked through the baking paper into the steam tray.

To this rice filled water pool was added 10 ml 3% peroxide.

The lid was placed ontop and the water allowed to soak through.

After 1 hr soaking the rice was moved to the microwave and steamed (in the mwsteamer) for 20 mins.

The hot rice was removed from the MW.

To my mates surprise it was almost solid again (dry despite the steam) .. too much heat ?

100 Ml water with 10 ml 3% peroxide was added to the rice and the lid replaced.

When cool inoculation of all experiments was peformed.

inside a hot oven 3 syringes were prepared from 1/4 glass spore print.

All 12 ml syringes had been mwaved with water and peroxide in them to steam, all tips were burnt in natural gas flame until all metal glowed and all water was pre boiled (taken from the pot the boiled the pfjars the night prior [ the lid was never opened until the syringe removal of the water - needle tip sterilised inbetween each water removal).

The pfjars each recieved 6 1ml injections 4 around outside and 2 in middle.

The pfcake was injected 6 times with 1ml spore solution as was the rice cake (which is still in the mw steamerdish with peroxide & water underneath [in steamtray] for extra humidity)

[ 30. March 2005, 14:18: Message edited by: electro ]

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cardboard spawn works very well for the woodloving species you might want to grow and yes ive heard b plus CAN use it though its not preferred

BTW when i referred to 'tubs' for the ferment tek i meant the big rolly bins ("rubbermaids")

interesting micro-teks

Microwaving cleaned syringes fully drawn with only a mL of peroxide water inside while in a zip lock bag is enough to sterlise (3 to 5 min on high)

I clean my needles by forcing hot water through them, then i boil them and drain, recap and wrap in foil. I pressure cook them but steaming them on high in a steamer basket on the stove for 15 mins is just fine as well

I tried a paper/bran microwave tek once but it failed.I tried inoculating with mycelial suspensions but the lack of sterility caused bacteria to reclaim the bran fraction (5%) the souring must have inhbited the fragments

funnily enough i then spawned it with a particularly vigorous wood mulch species via transferring a few pieces of colonised paper spawn to the top and resealing the lid (non-sterile).

the spawn i though was lost recovered and colonised very well it seems the mature and largere pieces of maycelium hasd the strength to overcome and consume the bacteria

- i then used it to inoculate some woodpiles where it is still moving fast

[ 15. November 2003, 23:58: Message edited by: reville ]

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Guest electro

updates:

WBS experiment ... no longer smells and is thriving in straw + dung outsdoor substrate .... mmmmmm mushie smells .... :)

5 day mark

Microrice 1.5l Peroxy tek

Pink AND green mould contamination in rice ... this definatley got there when extra water was added after sterilisation in an attempt to rehydrate the hardened steamed rice

Micropf 1.5l Peroxy tek

No aparentContamination

Slight (not fully positive yet) possible mycelial activity

PF Standard

No Contamination aparent

1 jar with definate mycelial growth

1 jar with possible mycelial growth

Peroxy PF

No Contamination aparent

2 jar with definate mycelial growth

1 jar with possible mycelial growth

Current temp 21.5c

4 x 1/2 hr portable fannings of container (to completley replace air 4 times a day)

after notations made .....

heater moved into a water bottle sitting in flat wide water tray along with all jars and microperoxypf experiment.

1 inch water filled

current temp approx 25-29 constant (though not measured yet)

[ 30. March 2005, 14:21: Message edited by: electro ]

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Guest electro

contamination source found -

upon fetching the spore print saddy to try to scrape together some "last hope" spores a 2 cm hole was discovered in it's lower side ... letting lovely contams in to bypass any sterilisation procedures applied to substrates --- the saddy was filled with peroxide, left sit for ten mins, then dumped into a sterilised peroxide malt jar ... hrm damn fragile plastic ! :/

New Updates:

Micropf 1.5l Peroxy tek

No aparentContamination

Mycelial activity

PF Standard

3 jar with definate ( 20 - 40 % colonised) mycelial growth

3 x Contaminated jars (100% failure) - Bluegreen mould - penacillium ? (down fall of being near a major pharma production area ?... antibiotic makings ????)

Peroxy PF

3 jar with definate ( 20 - 80 % colonised) mycelial growth

2 Contaminated jars (one only very slightly)

Current temp 24.5c

4 x 1/2 hr portable fannings of container (to completley replace air 4 times a day)

Anti mould experiment .....

The 6 jars were injected with more 3% peroxide in an attempt to give the mycellium a chance to beat the agressive bluegreen mould.

amounts of injected peroxide are given below

the non peroxie pf jars:

1:17ml (worst blue green infection)

2:15ml

3:14ml (3rd worst infection)

peroxie pf jars

1:10ml (slight infection)

2:5ml (3mm diameter infection)

3:5ml (no infection - added as insurance)

results:

Huge amounts of bubbling upon injections

Mycellial growth unaffected

blue green mould growth appears to have haulted in all cases, and being overgrown by fresh white mycellium in some cases [namely in the original peroxie pf jars])

conclusion: much more peroxide was needed initially to stop blue green mould appearing.

even more up to date results

1.5l mv pf cake colonising nicely - no contam, slow growth (but picking up)

all other cakes infected, bg mould picked up speed 2 days after peroxide treatment ... it won

some mycellium was transferred from the 1.5 l cake to herb jars filled with light malt and water (which had been boiled in a steam bath for 1 hr on highest gas flame available on stove) in an attempt to save the mycellial culture.

2 ml peroxide was added to 3 of the jars once cool (along with the mycellium) and 3 ml to the other two.

[ 28. November 2003, 12:16: Message edited by: electro ]

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"ingredient besides water is sodium laural sulphate which theoreticly kills bacteria "

SLS doesn't kill bacteria.

We use it to select for Enterobacteria, faecal coliforms, E.coli etc..

Kai.

If Laural equivalent to lauryl, otherwise just ignore.

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Guest electro

lol, that will "learn" him for trusting net sources .... heh

still, clearasil (which claims it is an antibacterial was used instead ..)

thx for the correction though.

updates

the final (1.5l) pf cake contaminated but not before healthy mycellium was transferred to peroxide malt jars.

2.5L (volume) of wet kitty litter was loaded into the 3l microwave steamer dish.

the dish was filled with water (covering all recycled cardboard kitty litter). To this water was added 1g condies crystals (potassium permangenate).

The soltion went bright purple then almost black as the crystals dissolved...

the cardboard was massaged so that the solution would be sucked up into the cardboard.

the solution was left for 20 mins then poured out leaving brownish purple cardboard (the purple turns brown as the KMNO4 oxidises the hell out of everything it touches ... the cardboard browns both through oxidation and the depositing of the non water soluble metal (manganese (the m) i think).

to the wet cardboard mush was added 5 desert spoons of light malt.

another .5 gram of permanganate was added to the mix and the dish refilled with water.

this water was stirred for 5 mins (to distribute the malt and permanganate evenly) then left for a futher 20 mins (after which time the purple had turned brown with malt and (manganese ? suspended in solution).

the excess liquid was poured off and the dish microwaved (with a little water in the water holder for the steamer) for 15 mins ... 10mins to get to boil, 5mins at boil.

while still untouchably hot the malty cardboard was transferred to a large glad snaplock bag and sealed in.

3 ml of peroxide was added to one of the malt liquid mycellium jars prepared several days earlier and the entire jar (150 ml max) dumped into the bag once the temp dropped to 35C.

(the jar has lots of white fluffy stuff creeping around all the sides esp on all floating chunks of pf mix [that was used to inoculate the jar])

the bag was transferred to an incubator (@25C) and left to see what happens.

[ 30. March 2005, 14:22: Message edited by: electro ]

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