Dear Casey:
I agree with your colleague that the FV3 ATCC isolate should be considered Biosafety Level I for the following reasons:
(1) This isolate is native to North America, and
(2) Ranaviruses are not zoonotic (which generally is reserved for BSL2 organisms unless they’re non-native and represent a significant risk to endemic wildlife).
I forwarded your email to the University of Tennessee Biosafety Officer (Brian Ranger); his response is below and he agrees with BSL-1 classification. He also mentions that it isn’t uncommon for different organizations to rank a pathogen at different biosafety levels based on perceived risk to humans or spillover to native wildlife populations. For example, colleagues in Europe might classify the ATCC FV3 strain as BSL2 in countries where it is not known to occur. At UT, Brian classifies all FV3-like ranaviruses as BSL1, but we follow BSL2 SOPs when working with Batrachochytrium salamandrivorans (Bsal), which is not known to occur in the USA. Brian thoughts on ATCC’s BSL2 classification are below, which may be a result of them exercising additional caution for liability purposes. Also, it is possible that ATCC could sell FV3 to consumers located outside of North America. Ranaviruses that infect amphibians (including FV3) are listed as notifiable by OIE. The biosecurity and isolation procedures you’re following are good and should be sufficient to ensure containment.
There may be others on the LISTSERV that have additional insight. If you have any questions regarding my thoughts, please do not hesitate to give me a call.
All the Best, Matt
______________________________
______________________________ __
Matthew J. Gray, Ph.D., ProfessorCo-chair, PARC National Disease Task Team
Chair, Research Working Group of the North American Bsal Task Force
Past Director, Global Ranavirus Consortium
Associate Director, UTIA Center for Wildlife HealthUniversity of Tennessee Institute of Agriculture (UTIA)
274 Ellington Plant Sciences BuildingKnoxville, TN 37996-4563
865.974.2740 [ofc] [log in to unmask]
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For more information about Forestry, Wildlife and Fisheries at UTK,
Please visit http://fwf.ag.utk.edu/or call 865.974.7126
____________________________________________________________ _
From: "Ranger, Brian Shayne" <[log in to unmask]>
Date: Thursday, March 29, 2018 at 4:42 PM
To: Matt Gray <[log in to unmask]>
Subject: RE: Ranavirus biosafety question
Hi Matt,
Welcome to the subjectivity of biosafety! Agents are often evaluated for risk from different perspectives, so the murky answer we always go to is ‘it depends’. I think in this case ATCC, as a public repository, has erred on the conservative side potentially due to concern related to ecological risk to indigenous fauna (not human risk). ATCC likely is encouraging good containment practices by setting the level at BSL-2; i.e., they want to ensure that they document that the materials are to be handled competently. When you look into their specifications sheet they basically outline good lab hygiene, wearing PPE, proper disinfection, and collection and inactivation of waste. They refer to the BMBL for other instructions, but do not specify primary containment devices/engineering controls or other specialized facility needs. ATCC does disclose in the specs that the agent is not a human hazard.
On my own soapbox, this is the result of trying to categorize bioagents into neat ‘buckets’ even though these are often highly subjective and don’t take into account the continuum of risk and/or multiple (and acceptable) approaches to risk mitigation. If I were to write a formal assessment on fv3 from indigenous sources, I would designate it as risk group 1/biosafety level 1 (with a few enhanced precautions…much like how you operate). If it were more exotic, and the risk to local fauna was deemed to be high/serious, then the stringency of practices and containment would increase.
Based on the description provided by your colleague on the listserv, I think his/her (?) approach is adequate. The shared lab does complicate the situation, but it looks like appropriate precautions (particularly dedicated PPE and traffic flow control) are in place.
Hope this helped some.
Brian S. Ranger, MS, SM(NRCM), CBSP
Biological Safety OfficerThe University of Tennessee, Knoxville
Office of Research & Engagement-Biosafety Program
UT Drive Services Bldg. A, Room 134
2233 Volunteer Blvd.
Knoxville, TN 37996-3000[log in to unmask]
o:(865) 974-1938c:(865) 230-0389
f:(865) 974-5416
http://biosafety.utk.edu
Big Orange. Big Ideas.
From: Gray, Matt
Sent: Thursday, March 29, 2018 2:16 PM
To: Ranger, Brian Shayne <[log in to unmask]>
Subject: FW: Ranavirus biosafety question
Brian:
Please see the question below. What BSL would you classify ranavirus? I don’t know why ATCC would classify FV3 (a species of ranavirus native to North America) as BSL 2. Thanks much for the feedback!
All the Best, Matt
From: "Finnerty, Casey M" <[log in to unmask]>
Date: Thursday, March 29, 2018 at 1:58 PM
To: Matt Gray <[log in to unmask]>
Subject: Ranavirus biosafety question
Dear Dr. Gray,
I posted the message below to the Global Ranavirus Consortium listserv, but I have not received any reply. If possible, I would very much appreciate hearing your thoughts on the two ranavirus biosafety questions I have below. I need to classify the BSL level with my department, and it would help to have your thoughts.
Thank you,
Casey Finnerty, Ph. D.
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. 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. Casey
Casey M. Finnerty, Ph. D.Associate ProfessorWinona State UniversityBiology Department - Pasteur 236Office: 507-457-5855Fax: 507-457-2599