Acacia phlebophylla TC contam disaster

[Darklight]

Just got back from a few days away, and as usual ran straight to the TC lab to check on any changes in my babies

Most things are fine. Some of the new species that got initiated into culture in the last batch are contam, and some aren't. You get that with new species in culture.

But I looked at the phlebs. And heartwrenchingly almost all of them have succumbed to a severe bacterial infestation. I'm so sad. Excuse me while I say "oh fuck" a whole lot. Is there an emoticon for copius weeping?

We were getting such good results. We were planning to deflask some and plant them out to try to establish a general deflasking protocol. We were planning to try to increase the multiplication rate. Now I can't be certain how many we have left, and how many parent plants this will leave us with- I'd consider 20 individual parent plants to be the minimum base for a good stable genetic program but it doesn't look like we even have that many

The work can continue but I'm not sure at what rate- certainly it will take us a year to get back to where we were, but at least now we have a working protocol.

The bacterial infestation is uniform in type and of plant origin ( starts at the base of TC plants on the media and works it way out ). It's also of a type that has been seen recurring in this species and nowhere else on any other species. It isnt the media sterilisation regime, not only is all contam of plant origin, but another species I am using an identical media for show no contamination at all.

From this I am trying to decide whether the contam came from a cultured plant early in the run and is pervasive and survives the rigorous sterilisation process I put forceps etc through to make its way through the rest of the batch, or that it is a systemic contam used by the species in vivio and pops up after time in all cultures when cultures are at a certain age

Re-sterilisation of plants isn't something I have found successful in TC. It's hard enough disinfesting parent plants where the contam isn't visible, by the time you can see a contam in TC its usually pretty rampant and re-disinfestation is harsh on the plants, has a high mortality rate, and is terribly laborious for nil reward in my experience.

Whatever it is, the sitch isn't looking good. We'll get back here, and maybe we can save a few of the older bacto plants. But shit it makes me miserable. I could weep

[planthelper]

i realy feel for you dl, and i wish you all the best.

let's cheer her up,

ogi, ogi, ogi, hoy, hoy, hoy,

ogi, hoy, ogi, hoy,

darklight you rock,

yeaaaahh....

my thought voice tells me phlebos dislike, the stagnant conditions inside flask's and ask for clean air movement around the plants.

[ 27. January 2005, 07:33: Message edited by: planthelper ]

[Stonehenge]

I'm sure any suggestion I make will be obvious and have already been considered, but what about using an antibiotic? Would it be too late to save some of the contaminated ones or might it help with future work?

Stoney

[Darklight]

Stonehenge:

but what about using an antibiotic? Would it be too late to save some of the contaminated ones or might it help with future work?

Shit Stoney, thanks for that. I can be such a bimbo... I do have a couple of antibiotics in stock and I'll look into their specific applications. They can apparantly be a bit harsh on plants, but hey not as harsh as bacto contam

Yer a goddamn legend there cob :D

[friendly]

Deepest condolences. I hope you can save some if not most of them.

[Darklight]

friendly:

Deepest condolences. I hope you can save some if not most of them.

Have found out what the problem is, it's a change in the media cooling method which caught up the contam biostat and sent it precipitating to the bottom with some other stuff. There were a few flasks which I hadn't done this to, and they're fine.

We have sufficient plant material so far to continue, and that's exactly what I'm doing today

[gerbil]

you're an absolute champion darklight, fantastic work!

It's good to hear things are working out for you, can't imagine how shattered i'd be in your situation, keep it up mate

[ 11. February 2005, 07:34: Message edited by: gerbil ]

[devance]

Silver Nitrate?

[Darklight]

Thought I'd better post that we got past the TC contam disaster.

Some seriously shonky advice from the ppl who installed the aircon unit caused hidjus amounts of contam to build up in the overflow. It was taking them months to send anyone out and they charged me a fair shitload to sort problems out, but this was way too critical to wait for

Fortunately the lovely mescalito, himself a recovering aircon dude of some note, came to our rescue with some excellent, timely and very accurate advice on what the problem was, how to fix it, and generally how to handle the aircon system overall. No idea if it was a coincidence or seasonal variation, but our power bill dropped by over 1/2 since the aircon was fixed

To give you an idea as to how bad the problem was, once I got around to sorting out the overflow tray, I found 8L of sludgy bacto water that had been sitting near the ceiling of my lab while I worked, and contaminating the fuck out of everything, which came pouring over my head once I shifted the tray to check it. Lots of language and panicking. And more language. But mesc's repair instructions were very clear and there hasn't even been any water sitting in the tray since ( and yes I do keep it clean now! )

So I'd like to say a big public and warm thanks to mesc, without whom everything would be dead and very smelly

OK now I have to go back in there and acutally do some work...

[ 20. June 2005, 11:48: Message edited by: Darklight ]

[hebrew]

thats what i like about this community, there are people with all sorts of knowledge and backgrounds and are willing to help each other out

on ya mesc!

[AndyAmine.]

Magic!

Three cheers for mesc!

Im so happy to hear things werent as bad as first thought.

[ 20. June 2005, 15:56: Message edited by: AndyAmine. ]

[Darklight]

AndyAmine.:

Im so happy to hear things werent as bad as first thought.

Actually they were worse than we first thought, the aircon was actively circulating contaminants in the transfer/ grow room which overloaded the media biostat's capacity to deal with them. In all species I'd worked on over a month. And if mesc hadn't helped I would have had to wait ages to get the aircon looked at. Mesc made it simple and it got sorted. :cool:

[nabraxas]

Three cheers for mesc!

I'll second that!!! :D

Well done people!!

[AndyAmine.]

Hmm, very lucky it was caught.

I was meaning the Phelb project in particular, that you didnt loose everything and have to start from scratch again.