Appendix A: Biosafety in the virology laboratory

Working with viruses which are infectious to humans poses a considerable risk of laboratory-acquired infection. This was demonstrated on numerous occasions in the past, and led to the establishment of a classification of viruses and other infectious agents into four groups depending upon the hazards to the laboratory worker. A full description of the historical development and current guidelines for biosafety can be found in Biosafety in Microbiological and Biomedical Laboratories edited by JW Richmond and RW McKinney, third edition, May 1993. Published by the US Department of Health and Human Services, Publication number (CDC) 93-8395. Here we reprint from that publication the summary description of recommended safety practices at each of the four levels (Table 1). Biosafety level 4 (maximum containment) laboratories are expensive to maintain and exist only in Australia, Canada, France, Germany, Russia, South Africa, the UK and the USA. For human blood-borne pathogens, biosafety precautions have been issued to health care and clinical laboratory workers under the term 'Universal Precautions' and these need to be observed for all work involving contact with potentially infectious blood and body fluids in addition to utilization of the appropriate biosafety level laboratory. A full description of universal precautions has been published. (Occupational Exposure to Bloodborne Pathogens, Final Rule, US Federal Register 56, 64175-64182, 1991). Since the eradication of smallpox, routine vaccination with vaccinia virus has ceased throughout the world. However work with vaccinia virus continues in many virology laboratories because of its value as a gene vector, e.g. in various vaccinia-based expression systems. In persons vaccinated within the previous ten years, work with vaccinia virus poses no special risk, and the US policy is that such work can be conducted at Biosafety Level 2 by appropriately vaccinated persons in a laboratory area with appropriate warning signs to prevent entry of unvaccinated persons. However, vaccination itself can be associated with significant adverse reactions, and in the UK and other European countries vaccination of laboratory workers is not advised (and is illegal in some countries, e.g. Germany). The danger to unvaccinated laboratory workers who use vaccinia virus is that they could become infected at an inappropriate site (e.g. the eye), with the possibility of a serious adverse reaction. The laboratory director may wish, therefore, to raise work with vaccinia virus to Biosafety Level 3, where such unvaccinated persons are involved. The following classification of vertebrate viruses potentially infectious for humans indicates the appropriate biosafety level (2, 3 or 4) at which work should be performed. Note that some viruses appear twice, depending upon the type of work being undertaken. In general, work involving virus concentration to high titre, production of aerosols, or virus infection of an animal host may require a higher biosafety level than that needed for other types of work. The laboratory director is responsible for appropriate risk assessment, and on occasion may decide to select a biosafety level higher than that recommended in the following classification.

Virology Methods Manual ISBN 0-12-465330-8

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Virology methods manual

Level Two

Aabahoyo Abras Abu Hammad Acado Acara Adeno-associated virus Adenoviruses Aguacate Alfuy Almpiwar Amapari Ananindeua Anhanga Anhembi Anopheles A Anopheles B Apeu Apoi Aride Arkonam Aroa Aruac Arumowot Astrovirus Aura Avalon B19 Bagaza Bahig Bakau Baku Bandia Bangoran Bangui Banzi Barmah Forest Barur Batai Batama Bauline Bebaru Belmont Benevides Benfica Betioga Bimiti Birao Bluetongue Boraceia

Botambi Boteke Bouboui Buffalopox Bujaru Bunyamwera Bunyip Burg E Arab Bushbush Bussuquara Buttonwillow Bwamba Cacao Cache valley California encephalitis Calmito Calovo Candiru Cape Wrath Capim Caraparu Carey Island Catu Chaco Chagres Chandipura Changuinola Charleville Chenuda Chikungunya (vs 131/25)* Chilibre Chobar Gorge CIo Mor Colorado tick fever Coronavirus Corriparta Cotia Cowbone Ridge Cowpox Coxsackie Csiro Village Cuiaba Cytomegalovirus D'Aguilar Dakar Bat Dengue I t Dengue 2t Dengue 3t Dengue 4t Dera Ghazi Khan

Eastern equine encephalitis t Edge Hill Entebbe bat Enteroviruses Epizootic haemorrhagic disease (of deer) Epstein-Barr Erve Eubenangee Eyach Flanders Fort Morgan Frijoles Gamboa Gan Gan Gomoka Gossas Grand Arbaud Great Island Guajara Guama Gumbo Limbo Hart Park Hazara Hepatitis A Hepatitis B Hepatitis C Hepatitis D Hepatitis E Hepatitis F Hepatitis G Highlands J Huacho Hughes Human herpesvirus Human papillomaviruses Icoaraci leri Ilesha Ilheus Influenza A Influenza B Influenza C Ingwavuma Inkoo Ippy Irituia Isfahan Itaporanga Itaqui Appendix A

358

Appendix A Jamestown Canyon Japanaut Jerry Slough Johnstone Atoll Joinjakaka Juan Diaz Jugra Junin (vs Candid #1) Jurona Jutiapa Kadam Kaeng Khoi Kaikalur Kaisodi Kamese Kammavan pettai Kannaman galam Kao Shuan Karimabad Karshi Kasba Kemerovo Kern Canyon Ketapang Keterah Keuraliba Keystone Kismayo Klamath Kokobera Kolongo Koongol Kotonkan Kowanyama Kunji n Kununurra Kwatta La Crosse La Joya Lagos bat Landjia Langat Lanjan Las Maloyas Latino Le Dantec Lebombo Lednice Lipovnik Lokern Lone Star Lukuni Lymphocytic choriomeningitis M'poko Madrid Maguari Mahogany Hammock Main Drain Malakal Manawa Manzanilla Mapputta Maprik Marco Marituba Marrakai Matariya Matruh Matucare Measles Melao Mermet Milkers nodule Mi natitlan Minnal Mirim Mitchell River Modoc Moju Molluscum contagiosum Mono Lake Montana myotis leukoencephalitis Moriche M osq ueiro Mossuril Mount Elgon bat Mumps Murutucu Mykines Navarro Nepuyo Ngaingan Nique Nkolbisson Nola Norwalk Ntaya Nugget Nyamanini Nyando O'nyong nyong Okhotskiy Okola Olifantsvlei Oliveros Orf Oriboca Ossa Pacora Pacui Pahayokee Palyam Parainfluenza Parana Pata Pathum Thani Patois Phnom-Penh bat Pichi nde Pixuna Point Poliovirus Pongola Ponteves Precarious Pretoria Prospect Hill Puchong Punta Salinas Punta Toro Qalyub Quaranfil Rabies virus Reoviruses Respiratory syncytial Restan Retroviruses, including human and simian immunodeficiency viruses (HIV-1, HIV-2, and SIV) Rhinoviruses Rift Valley fever (vs MP-12) Rio Bravo Rio Grande Ross River Rotaviruses Royal farm Rubella Sabo Saboya 359

Appendix A: Biosafety in the virology laboratory

Virology methods manual Saint Floris Sakhalin Salehabad San Angelo Sandfly fever (Sicilian) Sandfly fever (Naples) Sandjimba Sango Sathuperi Sawgrass Sebokele Seletar Sembalam Serra do Navio Shamonda Shark River Shuni Silverwater Simbu Simian hemorrhagic fever Sindbis Sixgun City Snow Mountain Snowshoe Hare Sokuluk Soldado Sororoca Spumavirus Stratford Sunday Canyon Tacaiuma Tacaribe Taggert Tahyna Tamiami Tanapox Tanga Tanjong Rabok Tataguine Tehran Tembe Tembusu Tensaw Tete Tettnang Thimiri Thottapalayam Tibrogargan Timbo Timboteua 360 Tindholmur Torovirus Toscana Toure Transmissible spongiform encephalopathiest (Creutzfeldt-Jakob, kuru, scrapie and related agents) Tribec Triniti Trivittatus Trubanaman Tsuruse Turlock Tyuleniy Uganda S Umatilla Umbre Una Upolu Urucuri Usutu Uukuniemi Vaccinia (vaccinated persons) Varicella-zoster Vellore Venezuelan equine encephalomyelitis (vs TC-83) *t Venkatapuram Vesicular stomatitis- Indiana Vesicular stomatitis - New Jersey Vinces Virgin River Wad Medani Walla Wanowrie Warrego Western equine encephalitis Whataroa Witwatersrand Wonga Wongorr Wyeomyia Yabapox Yaquinea Head Yatapox* Yellow fever (vs 17-D) Yogue Zaliv Terpeniya Zegla Zika Zingilamo Zirqa Level Three Adelaide River Agua Preta Aino Akabane Alenquer Almeirim Altamira Andasibe Antequera Araguari Aransas Bay Arbia Arboledas Babanki Batken Belem Berrimah Bhanja Bimbo Black Creek Canal Bobaya Bobia Bozo Buenaventura Cabassue Cacipacore Calchaqui Cananeia Caninde Chikungunya Chim Coastal Plains Cocal Connecticut Corfu Dabakala Dhori t Dobrava-Belgrade Douglas Dugbe Enseada Estero Real Everglades Flexal Appendix A

Appendix A Fomede Forecariah Fort Sherman Gabek Forest Gadgets Gully Garba Germiston Getah Gordil Hantaan Herpesvirus simiae (B virus) Human polyomaviruses (JC, BK) laco Ibaraki Ife Ingangapi Inini Israel Turkey meningitis issyk- kul Itaituba Itimirim Itupiranga Jacareacanga Jamanxi Japanese encephalitis Jari Junin Kairi Kedougou Khasan Kimberley Kindia Koutango Kyzylagach Lake Clarendon Llano Seco Louping III Lymphocytic choriomeningitis (infected animals) Macaua Mapuera Mayaro M boke Meaban M iddleburg Mobala Mojui Dos Compos Monte Dourado Mopeia Mucambo Munguba Murray Valley encephalitis Nairobi sheep disease Naranjal Nariva Nasoule Ndelle Ndumu Negis hi New Minto Ngari Ngoupe Nodamura North way Odrenisrou Omo Oriximina Oropouche Oru ngo Ouango Oubangui Oubi Ourem Palestina Para Paramushir Paroo River Peaton Perinet Pery Petevo Picola Piry Playas Powassan Pueblo Viejo Purus Puumala t Radi Razdan Resistencia Rift Valley fever Rochambeau Rocio Sagiyama Sal Vieja Salanga San Perlita San Juan Santa Rosa Santarem Saraca Saumarez Reef Sedlec Semliki Forestt Sena Madureira Seoul Sepik Sho kwe Sin Nombre Slovakia Somone Spipur Spo ndwen i St Louis encephalitis Tai Tamdy Telok Forest Termei I Thiafora Thogoto Tilligerry Tinaroo Tlacotalpan Tocio Tonate Ttinga Turuna Venezuelan equine encephalitis Vesicular stomatitis (infected animals) Wesselbron West Nile Xiburema Yacaaba Yaounde Yellow fever Yoka Yug Bogkanovac Level Four Absettarov t Black Creek Canal (infected animals) Congo-Crimean hemorrhagic fever Ebola Guanarito 361

Appendix A: Biosafety in the virology laboratory

Virology m e t h o d s manual Hanzalova Hypr Junin Kumlinge Kyasanur Forest disease Lassa Machupo Marburg Omsk hemorrhagic fever Rift Valley Fever (infected animals) Russian Spring-Summer encephalitis Sabia Sin Nombre (infected animals) Tick-borne encephalitis

* Vaccine Strain t Viruses marked with this dagger are classified in Europe either one level above (for Level Two) or below (for Level Three) the US levels.

Table

1 S u m m a r y of r e c o m m e n d e d biosafety levels for infectious agents

Biosafety Level

Agents
Not known to cause disease in healthy adults Associated with human disease, hazard=autoinoculation, ingestion, mucous membrane

Practices
Standard Microbiological Practices BCL-1 practice plus: 9 Limited access 9 Biohazard warning signs 9 Biosafety manual defining any needed waste decontamination or medical surveillance policies

Safety Equipment (Primary barriers)

None required

Facilities (Secondary Barriers)

BSL-1 plus: autoclave available

Primary barriers= Class I or ii BSCs or other physical containment devices used for all manipulations of agents that cause splashes or aerosols of infectious materials; PPEs; laboratory coats; gloves; face protection as needed Primary barriers= Class i or II BSCs or other physical containment devices used for all manipulations of agents; PPEs: protective lab clothing; gloves; respiratory protection as needed Primary barriers=All procedures conducted in Class IIi BSCs or Class I or II BSCs in combination with fullbody, air-supplied, positive pressure personnel suit

BSL-1 plus: autoclave available

Indigenous or exotic agents with potential for aerosol transmission; disease may have serious or lethal consequences

BSL-2 practice plus: 9 Controlled access 9 Decontamination of all waste 9 Decontamination of lab clothing before laundering 9 Baseline serum

BSL-2 plus: 9 Physical separation from access corridors 9 Self-closing, double door access 9 Exhausted air not recirculated 9 Negative airflow into laboratory BSL-3 plus: 9 Separate building or isolated zone 9 Dedicated supply/ exhaust, vacuum, and decon systems 9 Other requirements as outlined in the recommended text

Dangerous/exotic agents which pose high risk of life-threatening disease, aerosoltransmitted lab infections; or related agents with unknown risk of transmission

BSL-3 practices plus: 9 Clothing change before entering 9 Shower on exit 9 All material decontaminated on exit from facility

Based on 'Biosafety in Microbiological and Biomedical Laborator/es' with kind permission of the Editor, JW Richmond

362

Appendix A