Dear colleagues,

A colleague recently recommended I post some ranavirus questions I have to
this listserv.

With some collaborators I have been working with two ranaviruses, a local
isolate of largemouth bass virus (LMBV) and frog virus 3 (FV3) with my
undergraduate research students.

First, what do you consider the biosafety level of ranaviruses in general or
these viruses in particular to be? ATCC lists FV3 as BSL-2, but an
experienced ranavirus colleague says ranaviruses are BSL-1.

Second, what are your recommendations for biosafety/biocontainment? We  only
work with these viruses in cell culture (using FHM, BF-2 and occasionally
BHK-21 cells). Our titers are typically 10e4 to 10e5 TCID50/mL. All work is
done in a biocontainment hood that is disinfected with 2% bleach and 70%
ethanol before and after work. UV sanitizing lights are left on for 20 min.
after virus work as well. All glassware, spent media, and disposables
leaving the lab are autoclaved. Everyone working on cell culture washes
his/her hands upon entering and leaving the lab, wears nitrile or latex
gloves when working in the hood and a lab coat that remains in the lab.
Unfortunately, we do not have space for each researcher to have their own
research lab, so this lab is shared with two faculty who use it for several
weeks each semester for teaching. I also use the lab for teaching cell
culture techniques. That said, virus infected cultures are kept in a
separate incubator in the lab not used by my colleagues and their students.
Virus stocks are stored in a separate refrigerator as well. We keep
uninfected, highly susceptible cell lines in the same incubator as our
infected cultures, and we have never observed transfer of viral CPE from our
infected to our uninfected cultures.

Finally, I should add that my colleagues who share this lab for their
teaching maintain colonies of zebrafish and Ambystoma tigrinum (tiger
salamanders) on separate floors of the building. Other than the biosafety
measures described above and avoiding traffic from the cell culture lab to
their colonies, is there anything else we should do minimize risks to their
colonies? Thanks in advance for your help.