Phage outbreak and response measures

what we're doing is E. coli's
genetic
engineering bacteria. Some time ago even three batches were infected with
phage
, one batch a day, we do is 60T fermentation tank.
of phage outbreaks:
1, PH has been rising, will be relatively high (we normally 7.00, rebound to 7.8).
2 and DO will soon pick up, rising to more than 130%.
3, the fermentation liquid thinned, the OD value quickly decreased.
4, the mirror can not see any bacteria.
5, the outbreak time is earlier than once, are pre-fermentation.
phage outbreak is very fast, then 30 minutes or so, when we just moved the fermentation cycle is so fermentation cycle 3 to 5h, too terrible;
outbreak after the whole plant bleaching powder spraying, new Jieer spraying, formaldehyde fumigation, for several days in a row every day to do hygiene;
personal views:
phage outbreak cause analysis
1, when the workshop replaced with a new leader, did not pay attention to environmental health;
2, when the workshop also carried out transformation, health is only slightly done, for 4 months in a row;
3, workshop transformation, disinfection of the whole gas, some small exhaust are discharged in the workshop, but also directly discharged into space, resulting in the entire workshop humidity is
relatively large
, laboratory personnel are not trained in place, sampling directly discharged, without going through the waste tank directly discharged into the sampling tank;
6, the duty room staff also did not have training in place, for the treatment of foam sometimes leads to escape fluid,
7, just started infecting phages, did not attract enough attention, did not take appropriate measures;
people think that the prevention of phages is the first important health environment, as well as workshop personnel management, in strict accordance with
GMP
implementation. I hope we can all discuss the treatment of phages.
measures to be taken after the phage outbreak:
discontinuation, cleaning and disinfection of the room with sodium hypochloride, high temperature sterilization of all appliances or soaking with sodium hypochloride.
: effective chlorine concentration of 2500 ppm, greater than 30min;
shaker
fumigated with formaldehyde plus potassium permanganate, and then treated with ultraviolet radiation for half an hour;
If the concentration of phages in the air is also high, it can be fumigated with formaldehyde, 3g/m2, or with ultraviolet radiation, but be careful, as both methods can cause corrosion to the instrument. Water should also be treated with sodium hypochlorate. Air phage detection: double-layer flat, the lower layer is the production of bacteria or sensitive bacteria
culture
-base, the upper layer is nutritious broth soft
agar
, double-layer agar plate open cover put 30min, 30 degrees culture overnight. See if there are phage spots.
't work, we'll have to stop production. Or change to
strain of
, because phages have very good specificity. And the most fundamental is to develop laboratory safety norms, to eliminate the spread of live bacteria.