I have bacterial contamination in my incubator and I want anything that may help me to get rid of this contamination because it's bothering me in my culture or any work inside incubator.
hello Biswas,
Bacterial contaminations(except mycoplasma) are easy to eliminate.Just try to apply an anti-bacterial agent or an ethanol solution to desinfect your incubator surface.Fungus contamination is not to be excluded. You might check it also.
Hope it helps.
Best,
Hamadi
Hello Dr. Biswas, Have you done any culture to confirm its only bacterial infection?? If so, wipe the incubator with 70% ethanol (preferably 2-3 days continuously) and air dry. If you still notice the growth, go for a fumigation. Good Luck.
Is your contamination come from the incubator , from your BSC or your media and reagents ?
Are yu working with one or diffrenent cell lines that are contaminated ?
First, you have to decntaminate your incubator with 0.5% bleach or hydrogen peroxyde 0.5% fo at least 10 minutes, clean with alcohol after. You can do the same for your BSC.
And change your media and solutions.
If you have intracellular contamination of your cell line, you have to change it, or add antibiotics in your media.
Good luck
Do Fumigation with Potassium permanganate and Formaldehyde solution (In Glass Petri plate keep 20-30g of kmno4 plus add 5-10 ml Formaldehyde: Be-careful about fumes, keep in incubator chamber for 1-3 days : then you can do ur work without contamination
With Good Luck
Dear Friends,
Thanks a lot for all your suggestions.
Initially, we, too, had a feeling that our stocks might have got contaminated as we are dealing with a variety of strains at a time. But subsequently this was also ruled out.
My clean room, media and reagents are contamination free and that we have confirmed through various methodical culture process. We had kept a few nutrient media plates open in various parts of the room, biosafety cabinet and incubator. The plates placed in the clean room and biosafety cabinet did not grow any organism even after keeping them open for a week. But those plates which were placed inside the incubator had 1 single strain in all. So, finally, we have arrived at the conclusion that our incubator is infected with 1 certain species of bacteria which is infecting almost all our cultures.
I've already cleaned the incubator with ethyl alcohol and had kept the incubator running overnight at its highest temperature, assuming that the heat / temperature might eradicate the spores of the bacteria, but it did not happen so.
Possibly, I've to go with the Fumigation process as suggested by Dr. Sethi or follow Dr. Hafid's process of decontamination.
Thank you Friends once again....
Regards,
Sudeepta
Hello, I recommend iodine tincture for cleaning the incubator. Moreover, using antibiotics in medium, will prevent many contamination
Spray alcohol inner surface, all the racks after dismantling those parts. Then let it dry itself. Don't wipe. Give contact time alcohol to kill microbes. Next thing is to add CuSO4 into water pan of the incubator. You may add a small amt of disinfectant too.
37 degree C waterbath is another source of contaminations in labs. You can use 10 fold diluted bleach made from bleach (normally 5 %) for laundry. Just take 1 part of this mix with 9 parts of DW and use this water for your water bath. Since this is diluted, this will not be corrosive to S/S waterbath.
Some incubator has it own Decon button to press and kill bacteria. However, this decon reach upto 90 degree C only. If this function works, it is OK. If not, you may use H2O2 to disinfect the incubator. There are commercial mini apparatus to use H2O2 6% as disinfectant. Alternatively, as mentioned above, we can use K2MNO4 and formaldehyde 37%. However, I am not sure how to get rid of pungent smell later after fumigation.
- Alcoohol iis not effective for eliminating Gram+, mycobacteria, some yeat and Spores.
- Bleach at the standard concentration 5000 ppm is not effective against spores and endospores, You have to increase the contact time for some bacteria , but it's corrosive
-H2O2 (at 0.5)is more effective , We are using the Accel TB that contains accelerated particles. but is not effectiveagainst endospores, unless using a concentrated one at 6 % at at leat 6 hours contact. (Be aware to eliminates traces of H2O2 by cleaning with alcohol/water).
- fumigation wth paraformaldéhyde/potassium permanganate, with hydrogen peroxyd vapor, or with chlorine dioxide (We used all methods) is a hard way to get rid of contaminaton, it's more recommended for high risk group bacteria, aerosol contamination, room contaminaton, it done also for BSC or HEPA filters décontamination / certification in BSL-3 facilities...etc you can do it for at least overnight, and you have to seal the room (to prevent risk for others), and extract the air before working in the room. very toxic method and must bedone by a trained persons.
Do a simple method by desinfecting your incubator, if does not work, I will segguest to send this bacteria for identification, at least you will know the species and if it produces endospores ?
I do believe the focus on fungal spores and endospores might be where the problem lies. Both can be in the room air and within the incubator. Check the room for moist areas as the fungi grow there, and dust can often contain endospores. The sporocides might be the best cleaning agent.
You could wash the inside of the incubator with bleach or something similar, or use shortwave UV lights inside the incubator for a few hours.
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