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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 allinclusive.� 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 HYPERLINK "http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_0 2.htm" \l "_SECTION_V._FOOTNOTES" Sections V-C, V-D, V-E, and VF , Footnotes and References of Sections I through IV.� Further guidance on agents not listed in Appendix B may be obtained through:� HYPERLINK "http://www.cdc.gov/" \n 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; National Animal Disease Center, U.S. Department of Agriculture, Ames, Iowa 50010, Phone:� (515) 862-8258.� 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. Appendix B - Table 1. Basis for the Classification of Biohazardous Agents by Risk Group (RG) Risk Group 1 (RG1) Agents that are not associated with disease in healthy adult humans Risk Group 2 (RG2) Agents that are associated with human disease which is rarely serious and for which preventive or therapeutic interventions are often available Risk Group 3 (RG3) Agents that are associated with serious or lethal human disease for which preventive or therapeutic interventions may be available (high individual risk but low community risk) Risk Group 4 (RG4) Agents that are likely to cause serious or lethal human disease for which preventive or therapeutic interventions are not usually available (high individual risk and high community risk) Appendix B-I.�� Risk Group 1 (RG1) Agents RG1 agents are not associated with disease in healthy adult humans.� Examples of RG1 agents include asporogenic Bacillus subtilis or Bacillus licheniformis (see HYPERLINK "http://www4.od.nih.gov/oba/rac/guidelines_02/APPENDIX_C.htm" \l "_Appendix_C-IV-A._Exceptions" \n Appendix C-IV-A , Bacillus subtilis or Bacillus licheniformis Host-Vector Systems, Exceptions); adeno- associated virus (AAV) types 1 through 4; and recombinant 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 HYPERLINK "http://www4.od.nih.gov/oba/rac/guidelines_02/APPENDIX_C.htm" \l "_Appendix_C-II-A._Exceptions" \n 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. Appendix B-II.�� Risk Group 2 (RG2) Agents RG2 agents are associated with human disease which is rarely serious and for which preventive or therapeutic interventions are often available. Appendix B-II-A.�� Risk Group 2 (RG2) - Bacterial Agents Including Chlamydia --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, Cl. chauvoei, Cl. haemolyticum, Cl. histolyticum, Cl. novyi, Cl. septicum, Cl. tetani --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 --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 HYPERLINK "http://www4.od.nih.gov/oba/rac/guidelines_02/APPENDIX_B.htm" \l "_Appendix_B-III-A._Risk" Appendix B-III-A (RG3)) including M. avium complex, M. asiaticum, M. bovis BCG vaccine strain, M. chelonei, 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 --Rhodococcus equi --Salmonella including S. arizonae, S. cholerasuis, 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. parahemolyticus, V. vulnificus --Yersinia enterocolitica 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. peruvania, 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. cynomologi, 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 --Eastern equine encephalomyelitis virus --Venezuelan equine encephalomyelitis vaccine strain TC-83 --Western equine encephalomyelitis virus Arenaviruses --Lymphocytic choriomeningitis virus (non-neurotropic strains) --Tacaribe virus complex --Other viruses as listed in the reference source (see HYPERLINK "http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_0 2.htm" \l "_SECTION_V._FOOTNOTES" 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 HYPERLINK "http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_0 2.htm" \l "_SECTION_V._FOOTNOTES" \n Section V-C , Footnotes and References of Sections I through IV) Caliciviruses Coronaviruses Flaviviruses (Togaviruses) - Group B Arboviruses --Dengue virus serotypes 1, 2, 3, and 4 --Yellow fever virus vaccine strain 17D --Other viruses as listed in the reference source (see HYPERLINK "http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_0 2.htm" \l "_SECTION_V._FOOTNOTES" \n 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 HYPERLINK "http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_0 2.htm" \l "_Appendix_B-IV-D._Risk" \n 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 --Other tick-borne orthomyxoviruses as listed in the reference source (see HYPERLINK "http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_0 2.htm" \l "_SECTION_V._FOOTNOTES" \n Section V-C , Footnotes and References of Sections I through IV) Papovaviruses --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 HYPERLINK "http://www4.od.nih.gov/oba/rac/guidelines_02/APPENDIX_B.htm" \l "_Appendix_B-III-D._Risk" \n Appendix B-III-D , Risk Group 3 (RG3) - Viruses and Prions) and restricted poxviruses including Alastrim, Smallpox, and Whitepox (see HYPERLINK "http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_0 2.htm" \l "_SECTION_V._FOOTNOTES" 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 - laboratory adapted strains including VSV-Indiana, San Juan, and Glasgow Togaviruses (see Alphaviruses and Flaviviruses) --Rubivirus (rubella) Appendix B-III.��� Risk Group 3 (RG3) Agents 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 --Francisella tularensis --Mycobacterium bovis (except BCG strain, see HYPERLINK "http://www4.od.nih.gov/oba/rac/guidelines_02/APPENDIX_B.htm" \l "_Appendix_B-II-A._Risk" Appendix B-II-A , Risk Group 2 (RG2) Bacterial Agents Including Chlamydia), M. tuberculosis --Pasteurella multocida type B -"buffalo" and other virulent strains --Rickettsia akari, R. australis, R. canada, R. conorii, R. prowazekii, R. rickettsii, R, siberica, R. tsutsugamushi, R. typhi (R. mooseri) --Yersinia pestis 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 --Semliki Forest virus --St. Louis encephalitis virus --Venezuelan equine encephalomyelitis virus (except the vaccine strain TC-83, see HYPERLINK "http://www4.od.nih.gov/oba/rac/guidelines_02/APPENDIX_B.htm" \l "_Appendix_B-III-D._Risk" Appendix B-II-D (RG2)) --Other viruses as listed in the reference source (see HYPERLINK "http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_0 2.htm" \l "_SECTION_V._FOOTNOTES" \n 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 Flaviviruses (Togaviruses) - Group B Arboviruses --Japanese encephalitis virus --Yellow fever virus --Other viruses as listed in the reference source (see HYPERLINK "http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_0 2.htm" \l "_SECTION_V._FOOTNOTES" \n Section V-C , Footnotes and References of Sections I through IV) Poxviruses --Monkeypox virus Prions --Transmissible spongioform encephalopathies (TME) agents (Creutzfeldt-Jacob disease and kuru agents)(see HYPERLINK "http://www4.od.nih.gov/oba/rac/guidelines_02/NIH_Guidelines_Apr_0 2.htm" \l "_SECTION_V._FOOTNOTES" \n 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 Appendix B-IV.�� Risk Group 4 (RG4) Agents 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 --Machupo virus --Sabia Bunyaviruses (Nairovirus) --Crimean-Congo hemorrhagic fever virus Filoviruses --Ebola virus --Marburg virus Flaviruses (Togaviruses) - 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 Hemorrhagic fever agents and viruses as yet undefined Appendix B-V.��� Animal Viral Etiologic Agents in Common Use The following list of animal etiologic agents is appended to the list of human etiologic agents.� None of these agents is associated with disease in healthy adult humans; they are commonly used in laboratory experimental work. A containment level appropriate for RG1 human agents is recommended for their use.� For agents that are infectious to human cells, e.g., amphotropic and xenotropic strains of murine leukemia virus, a containment level appropriate for RG2 human agents is recommended. Baculoviruses Herpesviruses --Herpesvirus ateles --Herpesvirus saimiri --Marek's disease virus --Murine cytomegalovirus Papovaviruses --Bovine papilloma virus --Polyoma virus --Shope papilloma virus --Simian virus 40 (SV40) Retroviruses --Avian leukosis virus --Avian sarcoma virus --Bovine leukemia virus --Feline leukemia virus --Feline sarcoma virus --Gibbon leukemia virus --Mason-Pfizer monkey virus --Mouse mammary tumor virus --Murine leukemia virus --Murine sarcoma virus --Rat leukemia virus Appendix B-V-1.�� Murine Retroviral Vectors Murine retroviral vectors to be used for human transfer experiments (less than 10 liters) that contain less than 50% of their respective parental viral genome and that have been demonstrated to be free of detectable replication competent retrovirus can be maintained, handled, and administered, under BL1 containment. % ! > v | } � � ' A B � � � @ � � � � � � � ' ( � � d � ���뺴����笧 ��������������� e � f � � " � # hp � R W b c � � ������������뺴� h?0� CJ aJ h?0� CJ aJ hp � hp � 6 � h?0� 6 � hp � 6 � hp � h?0� 6 � h?0� 0J j h?0� U h?0� 6 �] � h?0� hp � CJ aJ h?0� CJ aJ h?0� h?0� CJ h?0� 5 �CJ OJ QJ \ �^J aJ aJ 3 hp � h?0� A B � � - e f � � � � � gdp � � � � � � $ If � y � � � � $ � � [$ \$ a$ gdp � � � [$ \$ gdp � gdp � $ � � [$ \$ a$ gdp � �I � � � N � � � � � T kd� $ $ If T �� �0 ��C 8" �������� � ���� � 6 � � � � � � � �������� � ���� �2� 2� x 4� a� �T $ If gdp � T kd $ $ If T �� �0 ��C 8 " �������� � ���� � 6 � � � � � � � �������� � ���� �2� 2� x 4� a� �T ' ( ; � � � � # � � � N B B B � � [$ \$ gdp � T kdV $ $ If T �� �0 ��C 8 " ���� � � 6 � � � � � � � � � � ���� �2� 2� x 4� a� �T $ If gdp � T kd� $ $ If T �s �0 ��C 8 " �������� � ���� � 6 � � � � � � � �������� � ���� �2� 2� x 4� a� �T # $ S T S T • � ^ _ � � � d � � � � x � � F � � � � � � � � � � � � � � � � � � � � � � � � � � � � gdp � � � [$ \$ gdp � # $ � � � � � � 4 5 D E G X \ r � � � � $ % ' 7 � � � � S T S T ~ • � � ] ^ _ x � � � � � � � � � � ; F b d w � � � � � � � � � ) 4 ? x ������������������ ���������������������������������������� �������������������� b c � v ' h?0� CJ h?0� 0J aJ j hp � h?0� CJ h?0� U aJ hp � h?0� 6 �] � h?0� hp � hp � CJ aJ Nx � � � � � � � � � � � � E F � � � � � � � � � K S [ • � � � � � � � � � � � # $ / 1 @ Y Z � � � � � � � !- �- �- �- �- �- �- �3 5 D F �� � � � � * 2 4 B D N Y t { � � � � ��������������������������������������� ���������������������������������������� ����������� hp � h?0� 0J j h?0� U h?0� - h?0� 6 �] �Z� � � � [ � � � � $ 1 �F � � D � � � � ! =! r! �! �! �! � � � � � � � � � � � � � � � � � � � � � � � � � � � �� �L�^�� `�L�gdp � gdp � � � � � � � ! ! ! <! =! q! r! �! �! �! �! �! �! �! 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