https://ntrs.nasa.gov/search.jsp?R=19680021536 2020-03-12T10:15:16+00:00Z Comprehensive Biological Protocol for the Lunar Sample Receiving Laboratory Manned Spacecraft Center National Aeronautics and Space Administration Houston, Texas Prepared under Contract NAS 9-6157 to Baylor University College of Medicint:, Houston, Texas Submitted June 16, 1967 INTRODUCTORY STATEMENT In June, 1966, members of the scientific community of the Baylor University College of Medicine contracted to prepare a biological quarantine protocol for the safe handling and study of lunar material to be returned to earth from early Apollo missions--such material, the spacecraft, and crew to be isolated initially in the Lunsr Receiving Laboratory, Manned Spacecraft Center, National Aeronautics and Space Administration, Houston, Texas. This document, and its abbreviated operational counterpart, wr i tten under contract NAS-9-6157, const itute a final protocol which has been modified extensively from a preliminary document subm it ted on December 16, 1966. The goals of the Lunar Receiving Laboratory are mu1 tiple and broadly encompass the scientific disciplines of geology, geophysics, chemistry, and biology. The purpose of this protocol is to define the biological studies which might reasonably fulfill the goals of the Bioscience Working Group of the Manned Space Science Coordinating Com- mittee, as set forth here: The prime purpose of the laboratory would be to provide a formal mechanism for testing appropriate representative 1unar samples for the possible presence of agents that might be infectious or toxic for man, animals, and plants. It should be the goal of this laboratory to provide safety clearance for lunar samples, if possible, within a period of approximately 30 days. I .--National Aeronautics and Space Administration: Summer Conference on ~unar-Explorat ion and Science, NASA-SP-88, p. 234, July, 1965. An overall statement of policy has been published recently (Science B:525, Feb. 3, 1967) and is appended to this document. In preparing a protocol which would fulfill these goals, we have had the benefit of the wise and introspective counsel of many contributovs, and the guidelines set forth here reflect the judgment of a cross section of our national scientific community. The protocol, while broad in scope, \;itenpts to explore in depth the effect of the lunar material upon i-- -._, plant and animal species about which a great deal is already known.) The protocol is flexible and lends itself to easy revision as more information is accumulated concerning the lun=+rsample and as bio- logical techniques improve during the implementation of the laboratory. The biological protocol haslee main elements, the first being crew microbiology; the second, in vitro attempts to culture microorgani sms from the lunar samp:e; and the third, the direct challenge of the lunar shnple in biological systemshrew microbiology concerns itself wi th . -- - -/I the establishment of pre-flight microbiology profiles and the subsequent review of alterations in flora encountered in crew personnel following return to earth. These studies will be performed in quarantine areas of the Lunar Receiving Laboratory and will be limited in duration to the time required to establish the nature of the microbial burden carried by the crew and the assurance of their freedom from communicable disease. Since it will be impossible to test the lunar sample on all but a few earth species, portions of the sample will be tested in representative members of all major taxa. We have utilized the concept of "unity within diversity" of biological processes on earth, and the careful selection of certain key species by the scientific community provides a broad-based spectrum for testing purposes.. There exists a small but finite probability that lunar substance? ;a! be injurious to organisms on earth. Such biological injury may be due to an inherent toxicity of the material or to the capability of such material to propagate itself in earth species. The toxic materials may be classified as follows: a. Redioactive materials from the moon itself. Early studies within the Lunar Receiving Laboratory will explore this possibility. b. Unknown inorganic polymers poss'bly containing silica, boron, and other inorganic elements. c. Deleterious low-molecular-weight'compounds acting as cellu!ar and metabolic poisons, mutagens, irritants, antimetabolites, or anti- vitamirs. d. Unknown metallo-organic compounds, effects on terrestrial organlsms unknown. The replicative materials may be classified as follows: a. Organisms (viral, bacterial, or fungal) taken to the moon, subjected to high-incident radiation, and returned unwittingly to earth. Such organisms may have mutated and have no counter- part on earth. This is an unlikely event. b. Plant materials of lunar origin capable of reproducing on earth as autotrophs or heterotrophs in nutrient media (artifical or natural, as in soi 1, ponds, oceans), resulting in natural ized forms producing deleterious effects by contact or competition. c. Xerophilic life forms of lunar origin using as protoplasmic materials elements found in terrestrial organisms such as carbon, hydrogen, oxygen, sulfur, and phosphorus. d. The exijtence of living matter on the moon at an organizational level above that of the small metazoa or metaphytes has a probability considerably less than that for unicellular organisms and is virtually excluded from consideration. If one accepts these fundamental policies and the constraints Imposed upon the laboratory by space and time limitations, there are left only three points which deserve discussion here in the way of clarification. These are the philosophy of the testing process itself, the nature of the internal controls to be employed, and the statistical approach to an evaluation of a heterogeneous, unknown mixture whose toxic or micro- biological potential is unknown. In regard to the philosophy of the testing procedures, it is safe to say that never before has there existed a facility with such ambitious and demanding goals as those presented by the Lunar Receiving Laboratory. The committee responsible for the preparation of this protocol stresses the need, therefore, for high professional standards in the conduct of these studies and has encouraged the Space Agency to avail themselves of the vast technical competence existing in laboratories throughout this country. When possible, it is to be hoped that the Agency will employ outside consultation at all steps. A testing procedure of this magni- tude and scope demands a degree of supervision and insight which will be challenging in the extreme. Since the testing procedures can be no better than the skill of those employing them, and in view of the diversity of species selected here, it is anticipated that the laboratory management will utilize to the fullest the sound and competent advice of the academic community and those federal agencies (u. S. Department of the Interior, U. S. Departmet,t of Agriculture, U. S. Pub1 ic Health Service) which have been instrumental in establishing the need for this laboratory. An important point which the committee would stress concerns the sequence of events in handling the lunar material. Collection, transport, receipt, and opening of the sample, as well as its mixing, aliquoting, and distribution, constitute a part of the general introduction to this protocol. In microbiological testing, however, it will be noted that we have set up a series of Itchallengestt to host organisms, this group of hosts representing higber and lower vertebrates, invertebrates, unicellular organisms and plants. It is anticipated that, along with the initial in vivo challenge and in vitro studies using classic microbiologic techniques, parallel --in vitro challenges will be made to both animal and plant cells growing in tissue cul ture. These ttobservationalt' steps are to be fol lowed by a secondary in vivo challenge as well as in vitro classic microbiological techniques, using organic a~:d inorganic media containing such added nutrients as might be suggested by the initial elemental and orjan:c analysis of the lunar sample. This temporal order, of initial followed by secondary chal- lenges, constitutes the critical part of the microbiological protocol. If replicating forms exist, this sequence offers the greatest promise for their detection. Every system described in this protocol has as an internal control the requirement that direct challenge of in vivo systems be conducted with both untreated and sterilized lunar material. Absolute double-barrier techniques are to be used throughout the laboratory; in some cases specific pathogen- free or germ-free species will be used, and in all In vitro sys;clls absolute stsrility of the system is mandatory. Carefully controlled trial runs of a11 systems should be begun fully one year in advance of receipt of the first lunar sample, and "unknown" terrestrial soil samples should be carried through all systems to Insure the technical competence of the laboratory faci- 1i ty. Since no one can predict at this time whether or not the lunat sample will be harmful to biological systems on earth, it is impossible to predict ' precisely how much meterial will be required to carry out the testing pro- cedures outlined here. In such a sea of ignorance, we have had to rely upon an almost empiric statistical approach, which can provide answers only on a contingency basis. We have had to assume, therefore, that the lunar sample, if it contains microorganisms at all, contains them at very low conceiitrations. Frcm such a base, we have used the next assun~ption, that these low concentrations exist either at "near negligible1' or at "detectable" levels. Such assumptions have allowed us to estimate a high and low quantity of material to be employed in many of the various direct in vivo challenges. In certain smaller species, such calculations cannot be used, since one is limited in the net amount of material which the species can tolerate. Such decisions will be difficult at best, and, again, the judgment of those work- ing in the field has been used as the ultimate guideline.