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DropDemSh1tz
Stranger

Registered: 09/05/06
Posts: 47
Last seen: 11 years, 2 months
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Questionable Liquid Culture Contamination. Day 3
#6406215 - 12/28/06 07:30 PM (18 years, 24 days ago) |
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Damnit. I already posted this but my post isn't coming up. So again. Possibly more brief.
I have never grown so I do not know what ANY of this is supposed to look like. And there aren't many pictures of the initial growth from liquid culture.
My Procedure: A few days prior to inoculation I created 3 Liquid culture jars. Orissa India, Tasmanian, A Strain. After day 2 Orissa and Taz showed no growth so I decided to throw them out and remake them. A strain by this time showed vigorous white looking growth. A majority of my jars were populated with this A strain liquid. The liquid was sterilized for 30 minutes at 20psi prior to being inoculated.
Over the last 3 days I have inoculated ~20 jars. Sterilized in batches of 6-7 at 20psi for 50-70 minutes. The jars have polyfil filters in the middle and 2-4 inoculation points covered with duct tape. This tape went through the sterilization process and was merely lifted then replaced when the inoculation hole was exposed. the substrate is either Brown rice with Flax meal or Rye Berries soaked for ~25 hours and cleaned using ~10 gallons of "PuR" water.
My needle sterilization was done as follows 1.) Take needle and heat to red hot using a torch along the entire length of the needle, almost to the plastic (last .5-1 cm or so). 2.) Needle cooled using an alcohol prep pad 3a.)Immediately put into inoculation hole 3b.) Put into liquid culture to pull out some myc then put into inoculation hole immediately 4a.) For Spores 1cc or so per jar was used (Orissa, taz, BPK, F+) 4b.) For Liquid 10cc or so were used per inoculation point 5.) Cover inoculation point w/ fresh duct tape
This was all done on the top of the stove. Stove off, but the range air sucker was on. I had cardboard on the sides to produce an upwards draft. All of the work was done as far back on the stove as possible so that the largest possible draft was in front of it. The tin foil on the top of the caps was not removed until just before the needle was sterilized. Tin foil was not replaced.
So now the meat.
All of my A strain culture jars are showing the same type of growth. This is day 1 or 2 for most of them. However all of the growth is tinged slightly green/blue. There is definitely white growth around this green area. The only areas that are greenish are the areas directly in contact with the mycelium liquid they were inoculated with.
Note, the other BR&F and Rye show only slight signs of growth at this point, 1-2 days into their cycle. So whatever it is was not endemic in the substrate prior to inoculation as the growth here is small and white, no green, which would lead me to believe the sterilization of the substrate was done properly.
I just went back and opened one of them up that I thought had the most vigorous growth. It was definitely green as green can be, slight tinge of blue. I wiped it lightly with a paper towel and the green came off. Looks like I am fucked w/ trich or something similar. It looked VERY similar to the powdery looking growth that gets on oranges at my grandmas. I think the initial syringe or the liquid was contaminated, however I followed the same procedure for the liquid that I did the other stuff. Possibly the honey I used was contaminated and it was not killed in the PC process, however the other two jars grew essentially nothing over the same time period? So thatโs probably not likely.
There are only a few possibilities
A.) The "hood" setup I used was ineffective and introduced something to my needles when they were being transferred from culture to spawn jars. The others got lucky since they were in the minority.
B.) Initial culture was contaminated. However this seems unlikely as the myc in the culture jar is growing very very white, with no green in it to speak of. Would it show in a liquid culture?
C.) This is an initial phase of the take over and it will fade and turn totally white like I am wanting...
Can someone with experience please elaborate?
I am feeling based on gut reactions that these jars are fuct. I think I am just going to preemptively make 12 more and just wait it out to see what happens with these and get some input from you guys.
I will make self-sealing innoc hole and poly fill filtered air hole ones for the next batch.
pictures to follow
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DropDemSh1tz
Stranger

Registered: 09/05/06
Posts: 47
Last seen: 11 years, 2 months
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Re: Questionable Liquid Culture Contamination. Day 3 [Re: DropDemSh1tz]
#6406968 - 12/29/06 01:02 AM (18 years, 23 days ago) |
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I just bought some more pint jars (others were quart). I am going to experiment with the A Strain solution again. See if the contamination arises once again. If it does I will determine the solution to be void. I am also going to sput 1cc into a pint jar to see if it goes through. I am also making a new solution that I will try out and if it yields the same result in new ones I will pronounce the syringe contaminated.
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hightimesreader
Half assed question asker


Registered: 07/18/06
Posts: 2,543
Loc: In the air conditioning
Last seen: 12 years, 9 months
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Re: Questionable Liquid Culture Contamination. Day 3 [Re: DropDemSh1tz]
#6412888 - 12/31/06 08:28 AM (18 years, 21 days ago) |
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Lol, breif? Well, lets see if I can help... Alright first off, don't let it make you think your contamed yet becuase without pics, I can't be sure but the flow hood is somtimes not a good thing to have on while doing needle work or transfering things becuase it moves spores around and if it happens to come out of airflow then it might land where you dont want it. A good idea that I usually go by is use a small bathroom that isnt used much if possible. Liberally apply lysol/bleach+water/oust w.e and then close the door with your stuff in there and the fan on. 5 minutes later, come back and turn off the fan and go in and do your buiseness with the door closed. Secondly, what does your stuff smell like? If it smells like anything other then what mushrooms smell like (you can smell packaged mushrooms from the grocery store if you need to) then it's contamed. Pictures would help diagnose your stuff better. Also, you might want to make it practice to use 15 psi for an hour to an hour and a half if possible. 30 min can work but it's also kind of sketchy. Hope it helps man!
-Hightimesreader
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RLmushrooms420
Stranger

Registered: 07/03/23
Posts: 17
Last seen: 1 year, 2 months
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Re: Questionable Liquid Culture Contamination. Day 3 [Re: DropDemSh1tz]
#28383345 - 07/03/23 08:59 AM (1 year, 6 months ago) |
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I canโt figure out how to start a thread so Iโm looking for a little help with liquid culture also
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SirPsycho
Purple Belt in Google-Fu



Registered: 01/01/20
Posts: 7,340
Loc: Rent free in your head
Last seen: 14 hours, 49 minutes
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Re: Questionable Liquid Culture Contamination. Day 3 [Re: RLmushrooms420]
#28383349 - 07/03/23 09:00 AM (1 year, 6 months ago) |
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There's a button labeled "post" at the top of the page here:
https://www.shroomery.org/forums/postlist.php/Board/2
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