The September 2009 revisions of the NIH Guidelines for Research Involving Recombinant DNA Molecules specify practices for constructing and handling: (i) recombinant deoxyribonucleic acid (DNA) molecules, and (ii) organisms and viruses containing recombinant DNA molecules. The following Appendices of the NIH Guidelines are of special interest:1.Appendix G – Physical Containment – Appendix G specifies physical containment for standard laboratory experiments and defines Biosafety Level 1 through Biosafety Level 4. For large-scale (over 10 liters) research or production, Appendix K (Physical Containment for Large Scale Uses of Organisms Containing Recombinant DNA Molecules) supersedes Appendix G. Appendix K defines Good Large Scale Practice through Biosafety Level 3 - Large Scale. For certain work with plants, Appendix P (Physical and Biological Containment for Recombinant DNA Research Involving Plants) supersedes Appendix G. Appendix P defines Biosafety Levels 1 through 4 - Plants. For certain work with animals, Appendix Q (Physical and Biological Containment for Recombinant DNA Research Involving Animals) supersedes Appendix G. Appendix Q defines Biosafety Levels 1 through 4 - Animals.2.Appendix I – Biological Containment (See Appendix E, Certified Host-Vector Systems) – Appendix I-I Levels of Biological Containment – In consideration of biological containment, the vector (plasmid, organelle, or virus) for the recombinant DNA and the host (bacterial, plant, or animal cell) in which the vector is propagated in the laboratory will be considered together. Any combination of vector and host which is to provide biological containment shall be chosen or constructed so that the following types of "escape" are minimized: (i) survival of the vector in its host outside the laboratory, and (ii) transmission of the vector from the propagation host to other non-laboratory hosts. The following levels of biological containment (host-vector systems) for prokaryotes are established. Appendices I-I-A through I-II-B describe levels of biological containment (host-vector systems) for prokaryotes. Specific criteria will depend on the organisms to be used.3.Appendix E. Certified Host-Vector Systems – While many experiments using Escherichia coli K-12, Saccharomyces cerevisiae, and Bacillus subtilis are currently exempt from the NIH Guidelines under Section III-F, Exempt Experiments, some derivatives of these host-vector systems were previously classified as Host-Vector 1 Systems or Host-Vector 2 Systems.4.Appendix K – Physical Containment for Large Scale Uses of Organisms Containing Recombinant DNA Molecules – Appendix K specifies physical containment guidelines for large-scale (greater than 10 liters of culture) research or production involving viable organisms containing recombinant DNA molecules. It shall apply to large-scale research or production activities as specified in Section III-D-6, Experiments Involving More than 10 Liters of Culture. It is important to note that this appendix addresses only the biological hazard associated with organisms containing recombinant DNA. Other hazards accompanying the large-scale cultivation of such organisms (e.g., toxic properties of products; physical, mechanical, and chemical aspects of downstream processing) are not addressed and shall be considered separately, albeit in conjunction with this appendix.All provisions shall apply to large-scale research or production activities with the following modifications: (i) Appendix K shall supersede Appendix G, Physical Containment, when quantities in excess of 10 liters of culture are involved in research or production. Appendix K-II applies to Good Large Scale Practice; (ii) the institution shall appoint a Biological Safety Officer if it engages in large-scale research or production activities involving viable organisms containing recombinant DNA molecules. The duties of the Biological Safety Officer shall include those specified in Section IV-B-3, Biological Safety Officer; (iii) the institution shall establish and maintain a health surveillance program for personnel engaged in large-scale research or production activities involving viable organisms containing recombinant DNA molecules which require Biosafety Level (BL) 3 containment at the laboratory scale. The program shall include: preassignment and periodic physical and medical examinations; collection, maintenance, and analysis of serum specimens for monitoring serologic changes that may result from the employee's work experience; and provisions for the investigation of any serious, unusual, or extended illnesses of employees to determine possible occupational origin.5.Appendix P – Physical and Biological Containment for Recombinant DNA Research Involving Plants – Appendix P specifies physical and biological containment conditions and practices suitable to the greenhouse conduct of experiments involving recombinant DNA-containing plants, plant-associated microorganisms, and small animals. All provisions of the NIH Guidelines apply to plant research activities with the following modifications:Appendix P shall supersede Appendix G (Physical Containment) when the research plants are of a size, number, or have growth requirements that preclude the use of containment conditions described in Appendix G. The plants covered in Appendix P include but are not limited to mosses, liverworts, macroscopic algae, and vascular plants including terrestrial crops, forest, and ornamental species.Plant-associated microorganisms include viroids, virusoids, viruses, bacteria, fungi, protozoans, certain small algae, and microorganisms that have a benign or beneficial association with plants, such as certain Rhizobium species and microorganisms known to cause plant diseases. The appendix applies to microorganisms which are being modified with the objective of fostering an association with plants.Plant-associated small animals include those arthropods that: (i) are in obligate association with plants, (ii) are plant pests, (iii) are plant pollinators, or (iv) transmit plant disease agents, as well as other small animals such as nematodes for which tests of biological properties necessitate the use of plants. Microorganisms associated with such small animals (e.g., pathogens or symbionts) are included.The Institutional Biosafety Committee shall include at least one individual with expertise in plant, plant pathogen, or plant pest containment principles when experiments utilizing Appendix P require prior approval by the Institutional Biosafety Committee.6.Appendix Q – Physical and Biological Containment for Recombinant DNA Research Involving Animals – Appendix Q specifies containment and confinement practices for research involving whole animals, both those in which the animal's genome has been altered by stable introduction of recombinant DNA, or DNA derived therefrom, into the germ-line (transgenic animals) and experiments involving viable recombinant DNA-modified microorganisms tested on whole animals. The appendix applies to animal research activities with the following modifications:Appendix Q shall supersede Appendix G (Physical Containment) when research animals are of a size or have growth requirements that preclude the use of containment for laboratory animals. Some animals may require other types of containment (see Appendix Q-III-D, Footnotes and References for Appendix Q). The animals covered in Appendix Q are those species normally categorized as animals including but not limited to cattle, swine, sheep, goats, horses, and poultry.The Institutional Biosafety Committee shall include at least one scientist with expertise in animal containment principles when experiments utilizing Appendix Q require Institutional Biosafety Committee prior approval.The institution shall establish and maintain a health surveillance program for personnel engaged in animal research involving v
