SOPs and special procedures for working with M13 phage?

[Tom Hodder]

Hi All,

One of our members is looking at working with M13 phage and recently put forward a proposal for a project. When it was discussed by the safety committee, several members mentioned its (phage in general) reputation of infecting everything, and needing to nuke the lab from orbit to get rid of it.

Having no personal experience of this, I decided to do a bit of research (*), but the material I found seems to be fairly split along the lines of "phage is bad, prepare to have all your stocks destroyed" and others who said to "Use standard aseptic technique, and it will be fine".

I did find a paper on the subject, several chapters in protocol manuals, and several threads of discussion on researchgate;
https://www.researchgate.net/post/Can_anyone_suggest_how_to_keep_my_cells_safe_from_phage

There were a couple of interesting comments, for example someone mentioned its resistance to autoclaving "At least for M13 it is essential since it resists autoclaving as normally used to sterilize media, glassware etc." (Ariane Toussaint · Université Libre de Bruxelles)

and this other one... "There is really only one phage that causes problems, phage T1, a classic siphophage (flexible tail) with ~50 kb dsDNA genome. All of phage biology gets a bad rap because of T1, which can spread as a microaerosol because it is dessication resistant. T1 is virtually impossible to eradicate if you keep growing E. coli cultures that are sensitive to it." (Ryland Fletcher Young · Texas A&M University)

I was wondering whether its even reasonable to work with M13 in a diybio general lab, (rather than a dedicated phage lab), and whether there were standard SOPs that could be applied to such things.

Many Thanks,

Tom

London Biohackspace

Notes (*)

http://2014.igem.org/Team:CU-Boulder/Notebook/Safety

M13 phage - 13 phage are biosafety level 1 viruses. Phage only infect bacteria so the risk to humans is negligible. The helper phagemid have a weakened packaging signal which reduces their replication efficiency, however, these phage could infect E. coli and other bacterial species containing the F’ episome if the phage escaped laboratory containment. M13 phage are not synthetic organisms; their environmental impact if they escaped laboratory containment is small due to their disrupted packaging signal so they would be quickly outcompeted by wild-type phage. Precautions such as hand washing and use of gloves were implemented to reduce risk to lab members. Proper disposal of samples was implemented to avoid environmental contamination. 

"Bacteriophage contamination: is there a simple method to reduce its deleterious effects in laboratory cultures and biotechnological factories?" http://jag.igr.poznan.pl/2004-Volume-45/1/pdf/2004_Volume_45_1-111-120.pdf

Gertrudis Rojas · Center of Molecular Immunology:

"My experience is with M13 filamentous phage. We have been performing phage display and E.coli expression for years without serious problems. We do not separate neither labs nor materials. Contamination risk can be managed with simple measures."

Everyone hates T1 phage

(Eric Keen · University of Miami )

" One caveat, though: the one phage I'd definitely avoid at all costs is phage T1 -- it's incredibly persistent in labs and could conceivably cause problems for months afterwards.  "