flask contamination treatment

hi guys,
i have a question for those who does orchid germinatin from seeds.

I sowed some phal seed, it propageted just fine. few months later protocons were replated and here week later i have a contamination with fungi in one jar and other type of fingi in other jar.
I tried to save them buy replating not contaminated portion into new jar. I treated contaminated portion with fungicide (not nessecary specific for those kind of fungi) and than replated to fresh jar.
And still i have some contamination.
yes i did not use laminar flow hood (just yesterday i decided to ask our other department at work if they do have that kind of hood. and they do have! if only i knew it before!), since its home project and its my first attempt.

so here are questions.
Could those young very little babies be saved by putting them in moss maybe or any other methods?! If to treat jar with conatmination with UV light in laminar hood will it kill fingi in the jar?

Any advised are welcome!

Interesting question Svetlana. I don't have experience in flasking but would certainly like to know. :bump:

I've been considering trying X-ing's myself. Probably die of old age by the time they flower but:dito:

I put some in moss... They died :(. One single one, biggest i put separately its still looks ok though.

Other jars got their UV portion. But i read it only kill bacteria etc on surface so probably it did not affect gungi incide the jars. Plus I'm not sure when they change bulbs in that hood....
So very little hope :(
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At least you are trying!

Gosh, I hope you can salvage something out of this! Maybe the big one will make it.

The single one in pot died too :(. Base got rotten even though tiny leaves were fine.

Other portion from other jar was washed with other antibacterial stuff and places in Petri dish, with no media at all. Lets see how long they will last.
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Sorry about that Svetlana. Keeping my fingers crossed for these to survive

I would also like to share my experience.

Several factors can contirbute to fungi, unclean utensils, workplace, airflow...the main culprit are the dust in the walls, ceiling, tables, etc.

In short dust are everywhere, that is why most are sowed under sterile condition, or workplace.

And how you sow, sterilze while sowing, and even capping can contribute to the success.

In my experience, when contamination have occured, saving the protocorns, are just futile....but i still do try to save them...but they usually die. Maybe by some chance you may have some luck...GOOD LUCK.

Replating contaminated protocorns, in my experience are just a waste of media and bottle.
(I have also did what you have done in saving before, but alas no success also)

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Sterilizing bottles/petri dish and utensils...after cleaning (i only use dishwashing liquid), you can just subject it to UV light (NOT THE SAME AS THE BLACK LIGHT) for half an hour to 1 hour, depending on the amount of utensils to be sterilize.

But be VERY CAREFUL In dealing with UV light, do not expose yourself directly for a very long time.

I have a homemade UV plastic box lined with aluminium foil (reflection), with outer switch.
After using it, i usually wait for 15-30min before opening it, so as not to be affected by the fresh heat inside the box.

I took classes for flasking seeds and my instructor has been a God-send. I recently had contamination that was also fungus. My instructor scooped out all of the protocorms and mixed a bottle of 100 mls water and added 1 ml straight Clorox and 1 drop of dish soap. Swirled them around for about 3 minutes. Then rinsed them by pouring them into a strainer and let the soapy water drain off, then rinsed very well with sterile water. Put them into a new sterile flask and added about 3 mls sterile water and a couple of drops of PPM (You can also get this from Phytotec under the name of PTC3) Saved my flask.