Cell contamination - Small rapidly moving black rods (Feb/08/2008 )

This has to be a bacterial problem. You described the contamination as quickly moving rods (bacillus). Mold doesn't move the way bacteria does and it's easy to see mold contamination as it will have a main foci of growth with tendrils moving out. You can see mold growing in the plate with the naked eye and it doesn't neccessarily make the media turbid the way bacteria does. Make sure you (or whoever cleaned) used 70% EtOH. I've found people who think if 70% is good, 100% is better...wrong! Try wiping everything down with a dilute bleach solution as well. You may be facing a bacteria that can put off spores. These can go into the air and hence into the filter (hood and incubator). Just talk to your boss about the $$$ going down the drain in cells, media, reagents, time, ect. versus the price of a new HEPA filter. I've also heard of contaimination getting into the air vents supplying the room with ventilation. You may need to clean these as well. Good luck and keep us posted!

There is a bacillus lab in the department... so that could be likely! Definitely cannot see the infection with the naked eye & from your description it definitely doesnt sound like mould. I cleaned everything down & I can guarantee that I used 70% ethanol. I also used virkon on some general surfaces. I'm pushing for the filter to be changed as there is definitely a lot of $$$ going down the drain. Since the I did a big clean of the whole cell culture room the extent of the contaminations has definitely eased off. BUT they are still some happening - myself & another colleauge are still getting random flasks going down the infection so the source is obviously still there.

I've attached a short video clip of a colleauges cells where the infection hasnt really established itself just yet. I apologize for the poor quality. If you look closely you can see the small black rods moving around rapidly.

Any ideas what they are?

If there are random flasks contaminating and not necessarily every flask you touch, then most likely there is something wrong with technique. I was having a horrible time with random contamination until I realized it was coming from my watch! My watch was not being covered by the gloves and there is a ton of yucky stuff in between the links (sorry..gross I know). Do you wear lab coats when culturing and if so have these been washed (with bleach)? If not, you might want to look into getting a couple long-sleeved lab coats that have elastic bands at the wrist. You also need to make sure that no one from the bacillus lab is entering that room. I hate it but my tissue culture room is also a storage room so I've actually caught (and yelled) at people carrying a bacterial culture into the room to get something from storage. Sounds like you are on the right track though. One last thought, and I know it's a somewhat paranoid approach, but do you have any reason to think there might be sabotage? I've had a run in myself. I was suspicious so I split my cells into two flask and put one in our incubator and another in a friend's lab's incubator. My incubator: contaminated. My friends: just fine. It's sad but there are people who feel threatened by success and will try to slow you down. Just a thought.

That's definately bacteria. Looks like two different types as well. Just a normal bacillus along with a spirillum and that's where your problem is. A spirillum contamination is just about as bad as yeast. These guys can get up and move..FAST! They can also travel very easily from one plate to another and are very difficult to get under control...especially if pen/strep resistant as you mentioned earlier. Just keep cleaning, do the best sterile technique possible and don't throw away contaminated flasks in the same room! Get these flasks out of the room as soon as you see/suspect contamination! Use dilute bleach solution to wipe everything down twice a day (opening and closing the tc room) until you have this under control.

Just saw the video. Its a bacterium.
#1 Add media to a plate with no cells.
#2 Add serum to a plate with no cells.
If no contamination it is your cells.
If both it is your technique.
I always force people to never wear lab coats OR talk (really)
in the tissue culture room.

Yep the little buggers sure can move cant they?! Disgusting!!!! Have been getting rid of contaminated flasks in the main lab (rather than cell culture room) from the get go but still they persist. I'll continue to wipe everything down with bleach solution twice a day... hopefully that'll obliterate them!

If any else has any other suggestions they'll be greatly appreciated as always!

Agree, they look like bacteria ...

Usually, the first thing I will check is the medium (with all addictives, that is the one I'm currently using) when I get contamination.