APPENDIX B. CLASSIFICATION OF HUMAN ETIOLOGIC AGENTS ON THE BASIS OF HAZARD

This appendix reflects the current state of knowledge and should be considered a resource document. Included are the more commonly encountered agents and is not meant to be all-inclusive. Information on agent risk assessment may be found in the Agent Summary Statements of the CDC/NIH publication, Biosafety in Microbiological and Biomedical Laboratories (see Sections V-C, V-D, V-E, and V-F, Footnotes and References of Sections I through IV. Further guidance on agents not listed in Appendix B may be obtained through: Centers for Disease Control and Prevention, Biosafety Branch, Atlanta, Georgia 30333, Phone: (404) 639-3883, Fax: (404) 639-2294; National Institutes of Health, Division of Safety, Bethesda, Maryland 20892, Phone: (301) 496-1357; Biosafety Manager, National Animal Disease Center, U.S. Department of Agriculture - ARS, Ames, Iowa 50010, Phone: (515) 337-7772.

A special committee of the American Society for Microbiology will conduct an annual review of this appendix and its recommendation for changes will be presented to the Recombinant DNA Advisory Committee as proposed amendments to the NIH Guidelines.

RG1 agents are not associated with disease in healthy adult humans. Examples of RG1 agents include asporogenic Bacillus subtilis or Bacillus licheniformis (see Appendix C-IV-A, Bacillus subtilis or Bacillus licheniformis Host-Vector Systems, Exceptions); adeno- associated virus (AAV - all serotypes); and recombinant or synthetic AAV constructs, in which the transgene does not encode either a potentially tumorigenic gene product or a toxin molecule and are produced in the absence of a helper virus. A strain of Escherichia coli (see Appendix C-II-A, Escherichia coli K-12 Host Vector Systems, Exceptions) is an RG1 agent if it (1) does not possess a complete lipopolysaccharide (i.e., lacks the O antigen); and (2) does not carry any active virulence factor (e.g., toxins) or colonization factors and does not carry any genes encoding these factors.

Those agents not listed in Risk Groups (RGs) 2, 3 and 4 are not automatically or implicitly classified in RG1; a risk assessment must be conducted based on the known and potential properties of the agents and their relationship to agents that are listed.

--Acinetobacter baumannii (formerly Acinetobacter calcoaceticus)
--Actinobacillus
--Actinomyces pyogenes (formerly Corynebacterium pyogenes)
--Aeromonas hydrophila
--Amycolata autotrophica
--Archanobacterium haemolyticum (formerly Corynebacterium haemolyticum)
--Arizona hinshawii - all serotypes
--Bacillus anthracis
--Bartonella henselae, B. quintana, B. vinsonii
--Bordetella including B. pertussis
--Borrelia recurrentis, B. burgdorferi
--Burkholderia (formerly Pseudomonas species) except those listed in Appendix B-III-A (RG3))
--Campylobacter coli, C. fetus, C. jejuni
--Chlamydia psittaci, C. trachomatis, C. pneumoniae
--Clostridium botulinum, C. chauvoei, C. haemolyticum, C. histolyticum, C. novyi, C. septicum, C. tetani
--Clostridium difficile - C.diff
--Coxiella burnetii - specifically the Phase II, Nine Mile strain, plaque purified, clone 4
--Corynebacterium diphtheriae, C. pseudotuberculosis, C. renale
--Dermatophilus congolensis
--Edwardsiella tarda
--Erysipelothrix rhusiopathiae
--Escherichia coli - all enteropathogenic, enterotoxigenic, enteroinvasive and strains bearing K1 antigen, including E. coli O157:H7
--*Francisella tularensis specifically *F. tularensis subspecies novicida [aka F. novicida], strain Utah 112; *F. tularensis subspecies holarctica LVS; *F. tularensis biovar tularensis strain ATCC 6223 (aka strain B38) *For research involving high concentrations, BL3 practices should be considered (see Appendix G-II-C-2. Special Practices (BL3)).
--Haemophilus ducreyi, H. influenzae
--Helicobacter pylori
--Klebsiella - all species except K. oxytoca (RG1)
--Legionella including L. pneumophila
--Leptospira interrogans - all serotypes
--Listeria
--Moraxella
--Mycobacterium (except those listed in Appendix B-III-A (RG3)) including M. avium complex, M. asiaticum, M. bovis BCG vaccine strain, M. chelonae, M. fortuitum, M. kansasii, M. leprae, M. malmoense, M. marinum, M. paratuberculosis, M. scrofulaceum, M. simiae, M. szulgai, M. ulcerans, M. xenopi
--Mycoplasma, except M. mycoides and M. agalactiae which are restricted animal pathogens
--Neisseria gonorrhoeae, N. meningitidis
--Nocardia asteroides, N. brasiliensis, N. otitidiscaviarum, N. transvalensis
--Pseudomonas aeruginosa
--Rhodococcus equi
--Salmonella including S. arizonae, S. choleraesuis, S. enteritidis, S. gallinarum-pullorum, S. meleagridis, S. paratyphi, A, B, C, S. typhi, S. typhimurium
--Shigella including S. boydii, S. dysenteriae, type 1, S. flexneri, S. sonnei
--Sphaerophorus necrophorus
--Staphylococcus aureus
--Streptobacillus moniliformis

--Streptococcus including S. pneumoniae, S. pyogenes
--Treponema pallidum, T. carateum
--Vibrio cholerae, V. parahaemolyticus, V. vulnificus
--Yersinia enterocolitica
--Yersinia pestis specifically pgm(-) strains (lacking the 102 kb pigmentation locus) and lcr(-) strains (lacking the LCR plasmid)

Appendix B-II-B. Risk Group 2 (RG2) - Fungal Agents

--Blastomyces dermatitidis
--Cladosporium bantianum, C. (Xylohypha) trichoides
--Cryptococcus neoformans
--Dactylaria galopava (Ochroconis gallopavum)
--Epidermophyton
--Exophiala (Wangiella) dermatitidis
--Fonsecaea pedrosoi
--Microsporum
--Paracoccidioides braziliensis
--Penicillium marneffei
--Sporothrix schenckii
--Trichophyton

Appendix B-II-C. Risk Group 2 (RG2) - Parasitic Agents

--Ancylostoma human hookworms including A. duodenale, A. ceylanicum
--Ascaris including Ascaris lumbricoides suum
--Babesia including B. divergens, B. microti
--Brugia filaria worms including B. malayi, B. timori
--Coccidia
--Cryptosporidium including C. parvum
--Cysticercus cellulosae (hydatid cyst, larva of T. solium)
--Echinococcus including E. granulosis, E. multilocularis, E. vogeli
--Entamoeba histolytica
--Enterobius
--Fasciola including F. gigantica, F. hepatica
--Giardia including G. lamblia
--Heterophyes
--Hymenolepis including H. diminuta, H. nana
--Isospora
--Leishmania including L. braziliensis, L. donovani, L. ethiopia, L. major, L. mexicana, L. peruviana, L. tropica
--Loa loa filaria worms
--Microsporidium
--Naegleria fowleri
--Necator human hookworms including N. americanus
--Onchocerca filaria worms including, O. volvulus
--Plasmodium including simian species, P. cynomolgi, P. falciparum, P. malariae, P. ovale, P. vivax
--Sarcocystis including S. sui hominis
--Schistosoma including S. haematobium, S. intercalatum, S. japonicum, S. mansoni, S. mekongi
--Strongyloides including S. stercoralis
--Taenia solium
--Toxocara including T. canis
--Toxoplasma including T. gondii
--Trichinella spiralis
--Trypanosoma including T. brucei brucei, T. brucei gambiense, T. brucei rhodesiense, T. cruzi
--Wuchereria bancrofti filaria worms

Appendix B-II-D. Risk Group 2 (RG2) - Viruses

Adenoviruses, human - all types

Alphaviruses (Togaviruses) - Group A Arboviruses
--Chikungunya vaccine strain 181/25
--Eastern equine encephalomyelitis virus
--Venezuelan equine encephalomyelitis vaccine strains TC-83 and V3526
--Western equine encephalomyelitis virus

Arenaviruses
--Junin virus candid #1 vaccine strain
--Lymphocytic choriomeningitis virus (non-neurotropic strains)
--Tacaribe virus complex
--Other viruses as listed in the reference source (see Section V-C, Footnotes and References of Sections I through IV)

Bunyaviruses
--Bunyamwera virus
--Rift Valley fever virus vaccine strain MP-12
--Other viruses as listed in the reference source (see Section V-C, Footnotes and References of Sections I through IV)

Caliciviruses

Coronaviruses

Flaviviruses - Group B Arboviruses
--Dengue virus serotypes 1, 2, 3, and 4
--Japanese encephalitis virus strain SA 14-14-2
--Yellow fever virus vaccine strain 17D
--Other viruses as listed in the reference source (see Section V-C, Footnotes and References of Sections I through IV)

Hepatitis A, B, C, D, and E viruses

Herpesviruses - except Herpesvirus simiae (Monkey B virus) (see Appendix B-IV-D, Risk Group 4 (RG4) - Viral Agents)
--Cytomegalovirus
--Epstein Barr virus
--Herpes simplex types 1 and 2
--Herpes zoster
--Human herpesvirus types 6 and 7

Orthomyxoviruses
--Influenza viruses types A, B, and C (except those listed in Appendix B-III-D, Risk Group 3 (RG3) - Viruses and Prions)
--Tick-borne orthomyxoviruses

Papilloma viruses
--All human papilloma viruses

Paramyxoviruses
--Newcastle disease virus
--Measles virus
--Mumps virus
--Parainfluenza viruses types 1, 2, 3, and 4
--Respiratory syncytial virus

Parvoviruses
--Human parvovirus (B19)

Picornaviruses
--Coxsackie viruses types A and B
--Echoviruses - all types
--Polioviruses - all types, wild and attenuated
--Rhinoviruses - all types

Poxviruses - all types except Monkeypox virus (see Appendix B-III-D, Risk Group 3 (RG3) - Viruses and Prions) and restricted poxviruses including Alastrim, Smallpox, and Whitepox (see Section V-L, Footnotes and References of Sections I through IV)

Reoviruses - all types including Coltivirus, human Rotavirus, and Orbivirus (Colorado tick fever virus)

Rhabdoviruses
--Rabies virus - all strains
--Vesicular stomatitis virus non exotic strains: VSV-Indiana 1 serotype strains (e.g. Glasgow, Mudd-Summers, Orsay, San Juan) and VSV-New Jersey serotype strains (e.g. Ogden, Hazelhurst)

Rubivirus (Togaviruses)
--Rubella virus

Gastroenteritis causing viruses
Note: These were not on the NIH Guideline list.
The first 2 are on the Macquarie University (Australia) list
--Noroviruses
--Sapporo-like Viruses
--Group A Rotavirus

RG3 agents are associated with serious or lethal human disease for which preventive or therapeutic interventions may be available.

Appendix B-III-A. Risk Group 3 (RG3) - Bacterial Agents Including Rickettsia

--Bartonella
--Brucella including B. abortus, B. canis, B. suis
--Burkholderia (Pseudomonas) mallei, B. pseudomallei
--Coxiella burnetii (except the Phase II, Nine Mile strain listed in Appendix B-II-A, Risk Group 2 (RG2) - Bacterial Agents Including Chlamydia)
--Francisella tularensis (except those strains listed in Appendix B-II-A, Risk Group 2 (RG2) - Bacterial Agents Including Chlamydia)
--Mycobacterium bovis (except BCG strain, see Appendix B-II-A, Risk Group 2 (RG2) - Bacterial Agents Including Chlamydia), M. tuberculosis
--Orientia tsutsugamushi (was R. tsutsugamushi)
--Pasteurella multocida type B -"buffalo" and other virulent strains
--Rickettsia akari, R. australis, R. canada, R. conorii, R. prowazekii, R. rickettsii, R, siberica, R. typhi (R. mooseri)
--Yersinia pestis (except those strains listed in Appendix B-II-A, Risk Group 2 (RG2) - Bacterial Agents Including Chlamydia)

Appendix B-III-B. Risk Group 3 (RG3) - Fungal Agents

--Coccidioides immitis (sporulating cultures; contaminated soil)
--Histoplasma capsulatum, H. capsulatum var. duboisii

Appendix B-III-C. Risk Group 3 (RG3) - Parasitic Agents

None

Appendix B-III-D. Risk Group 3 (RG3) - Viruses and Prions

Alphaviruses (Togaviruses) - Group A Arboviruses
--Chikungunya virus (except the vaccine strain 181/25 listed in Appendix B-II-D Risk Group2 (RG2) - Viruses)
--Semliki Forest virus
--St. Louis encephalitis virus
--Venezuelan equine encephalomyelitis virus (except the vaccine strains TC-83 and V3526, see Appendix B-II-D (RG2) - Viruses)
--Other viruses as listed in the reference source (see Section V-C, Footnotes and References of Sections I through IV)

Arenaviruses
--Flexal
--Lymphocytic choriomeningitis virus (LCM) (neurotropic strains)

Bunyaviruses
--Hantaviruses including Hantaan virus
--Rift Valley fever virus

Coronaviruses
--SARS-associated coronavirus (SARS-CoV)
--Middle East respiratory syndrome coronavirus (MERS-CoV)

Flaviviruses - Group B Arboviruses
--Japanese encephalitis virus (except those strains listed in Appendix B-II-D Risk Group2 (RG2) - Viruses)
--West Nile virus (WNV)
--Yellow fever virus
--Other viruses as listed in the reference source (see Section V-C, Footnotes and References of Sections I through IV)

Orthomyxoviruses
-- Influenza viruses 1918-1919 H1N1 (1918 H1N1), human H2N2 (1957-1968), and highly pathogenic avian influenza H5N1 strains within the Goose/Guangdong/96-like H5 lineage (HPAI H5N1).

Poxviruses
--Monkeypox virus

Prions
--Transmissible spongiform encephalopathies (TSE) agents (Creutzfeldt-Jacob disease and kuru agents)(see Section V-C, Footnotes and References of Sections I through IV, for containment instruction)

Retroviruses
--Human immunodeficiency virus (HIV) types 1 and 2
--Human T cell lymphotropic virus (HTLV) types 1 and 2
--Simian immunodeficiency virus (SIV)

Rhabdoviruses
--Vesicular stomatitis virus (except those strains listed in Appendix B-II-D Risk Group2 (RG2) - Viruses)

RG4 agents are likely to cause serious or lethal human disease for which preventive or therapeutic interventions are not usually available.

Appendix B-IV-A. Risk Group 4 (RG4) - Bacterial Agents

None

Appendix B-IV-B. Risk Group 4 (RG4) - Fungal Agents

None

Appendix B-IV-C. Risk Group 4 (RG4) - Parasitic Agents

None

Appendix B-IV-D. Risk Group 4 (RG4) - Viral Agents

Arenaviruses
--Guanarito virus
--Lassa virus

--Junin virus (except the candid #1 vaccine strain listed in Appendix B-II-D Risk Group2 (RG2) - Viruses)
--Machupo virus
--Sabia

Bunyaviruses (Nairovirus)
--Crimean-Congo hemorrhagic fever virus

Filoviruses
--Ebola virus
--Marburg virus

Flaviruses - Group B Arboviruses
--Tick-borne encephalitis virus complex including Absetterov, Central European encephalitis, Hanzalova, Hypr, Kumlinge, Kyasanur Forest disease, Omsk hemorrhagic fever, and Russian spring-summer encephalitis viruses

Herpesviruses (alpha)
--Herpesvirus simiae (Herpes B or Monkey B virus)

Paramyxoviruses
--Equine Morbillivirus (Hendra virus)

Hemorrhagic fever agents and viruses as yet undefined Source Appendex B