April 1996
Although not explicitly prohibited by the 1972 Biological Weapons Convention (BWC), the use of biological or toxin weapons by States Parties would constitute prima facie evidence of a breach of the Convention. Implicitly recognizing this, States Parties to the BWC have called for "measures for the investigation of alleged use" as part of a "coherent regime to enhance the effectiveness of and improve compliance with the Convention...to be included, as appropriate, in a legally binding instrumentŠ" (Special Conference of Parties to the Biological Weapons Convention, September 19-30, 1994: Part II, Final Declaration, paragraphs 35-6). An effective mechanism for investigating alleged use of biological and toxin weapons will not only enhance compliance with the BWC by deterring use, but will also assure States Parties that any suspicious incident occurring on their territory will be investigated at their request. It will also offer a means by which countries wrongly suspected of violation can demonstrate their compliance, and it will discourage unfounded and destabilizing accusations.
An ad hoc and unfunded mechanism currently exists for investigating the possible use of biological and toxin weapons, as violations of the Geneva Protocol. The UN Secretary-General has been authorized by the General Assembly (Resolution 42/37 C, 1987, and Resolution 43/74 A, 1988) to maintain lists of qualified experts and laboratories provided by Member States for the purpose of developing technical guidelines and procedures for investigations (issued in 1989 in Report of the Secretary-General, A/44/561) and for undertaking such investigations. However, the BWC Special Conference implicitly recognized the need for a more formal and comprehensive mechanism, with its call for measures to investigate alleged use of BW as part of a legally-binding instrument or Protocol to the BWC. Such a mechanism will require a legally binding commitment by States Parties to cooperate with investigations, and an office with a permanent, international staff to provide the leadership, backup, and services needed for rapid and effective deployment of investigation teams, evaluation of the data collected, and preservation of confidentiality. It will be important for States Parties to the Protocol to make the mechanism they adopt available to the UN Secretary General for his use in any investigation of the possible use of biological or toxin weapons that he might undertake under the authority of the General Assembly resolutions.
With the aim of contributing to the development of an effective mechanism by the Ad Hoc Group, the Federation of American Scientists Working Group on Biological Weapons Verification constituted a Subgroup on Investigation of Alleged Use or Release of Biological or Toxin Weapons Agents, composed of technical experts on biological weapons control, molecular biology, microbiology, and epidemiology.
Literature Reviewed and Experts Consulted
The Subgroup has reviewed the technical literature on the subject of investigation of use of biological or chemical weapons, and on the general conduct of epidemiological investigation of human, plant, and animal disease. Special attention was given to publications analyzing the contributions epidemiology could make to the control of biological weapons,1 and to previous proposals for specific mechanisms for the investigation of chemical, biological, or toxin weapons use, most notably: the 1994 CWC;2 the 1989 report of the Secretary General of the United Nations on Chemical and Bacteriological (Biological) Weapons;3 and two publications of the Government of Canada.4
An earlier draft of this Report was sent to a number of experts around the world for review; their comments have been considered in the drafting of the final version.
Reporting of Unusual Outbreaks of Disease
Unusual outbreaks of disease most often have fully natural causes, despite their unusual attributes. However, because such outbreaks could also result from the use or release of biological weapons agents, and because even fully natural outbreaks might provoke suspicions of non-compliance, States Parties to the BWC have agreed that information on unusual outbreaks should be exchanged as a confidence-building measure. However, experience with this confidence-building measure makes it clear that States Parties may be reluctant to report outbreaks in connection with the BWC, even when they have willingly reported to the World Health Organization (WHO) or other international health organization.
A Protocol to the BWC should strengthen the reporting expectation by incorporating a legally-binding requirement to report unusual outbreaks, and it should specify that the reports should be directed to an appropriate international health organization rather than to an arms control organization. Reporting of human, animal, and plant outbreaks should be required; examples of appropriate international health organizations would include the WHO, the Food and Agriculture Organization (FAO), and the Office International des Épizooties (OIE).
Official reports of outbreaks received by international health organizations are publically accessible and would be available to States Parties and to the office established to coordinate the implementation of a legally-binding Protocol to the BWC, which should compile and analyze them.
Recommendation 1: The Protocol should require, on a legally-binding basis, that States Parties provide prompt and detailed reports on all unusual outbreaks of disease on their territories. Reports should be directed to the WHO, the OIE, the FAO or other appropriate international health organizations.
The prompt reporting of unusual outbreaks of disease to the appropriate international health agencies will significantly contribute to the understanding of global patterns of infectious disease, as well as to building confidence in the natural etiology of reported outbreaks. However, in rare cases there may be reason to question natural causation, and to suspect that a particular outbreak is the result of deliberate use or of accidental release during the development, testing, production, transfer, or stockpiling of biological weapons agents. Under such conditions an investigatory mechanism is needed. WHO, OIE, and FAO cannot be asked to conduct investigations of alleged BW use or release; their mandate is purely humanitarian and they would be unacceptably politicized if they were to undertake investigation of compliance with a disarmament treaty. A new mechanism and organization are needed for the investigation of suspect outbreaks (and only for suspect outbreaks; the BWC and its Protocol have no legitimate role in the investigation of outbreaks not suspected to be the result of use or release of biological weapons agents).
The BWC prohibits the acquisition of biological or toxin weapons agents that has no peaceful or prophylactic purpose. As noted above, it does not directly prohibit their use (a prohibition of use in war being previously established by the 1925 Geneva Protocol, and by customary international law as well). Investigation of biological weapons use as part of a Protocol to the BWC rests on the premise that the use of biological or toxin agents as weapons would constitute prima facie evidence for prior acquisition of agents that had no justification for prophylactic, protective, or other peaceful purposes, as well as the means for their delivery. This premise should be made explicit by States Parties at the Fourth Review Conference in their discussion of Article 1.
States Parties to the BWC are engaged in drafting a legally binding Protocol, to include a regime to enhance compliance. As noted above, the September 1994 Special Conference of Parties to the Biological Weapons Convention directed that measures for the investigation of alleged use of biological weapons agents should be included in the regime. In Appendix I we suggest language for this purpose. We believe that it will be necessary for the Protocol to establish an international office to administer and coordinate the measures agreed upon. The name of this office and the title of its administrative head are yet unspecified; for the purposes of discussion, that office is herein termed the "Biological Weapons Control Office" (BWCO), and assumed to be headed by a Director-General. However, the substance of our recommendations is independent of the name chosen and of the structural details of the office as it may ultimately be established.
Recommendation 2: The Protocol should contain a section establishing a mechanism for the investigation of alleged use or release of biological or toxin weapons agents.
The treaty language proposed in Appendix 1 of this report provides language for consideration, based on the comparable section of the CWC. It is, of necessity, rather general. It has to cover a very wide range of circumstances, and it must not be made obsolete by changes in technical capabilities or epidemiological knowledge. It can not (and should not) cover most of the operational details of the program. These will have to be developed by the BWCO as operating guidelines to implement the Protocol. The operating guidelines are expected to be dynamic, changing as experience with the process accumulates and as scientific advances make possible new methods of investigation.
Initiation of an Investigation
The range of possible uses of biological weapons agents provides an indication of the types of incident that the BWCO might have to investigate. These include: (1) use in overt war between states; (2) use in a civil war; (3) use by a government against an ethnic or religious group, or against an individual; (4) use by a state against another state in the absence of overt war; and (5) use by terrorists. In addition to such incidents of actual hostile use, biological weapons agents may be accidentally (or even intentionally) released during their development, testing, production, transportation, or stockpiling. In all of these cases, the agent may target plants, domestic animals, or humans. Use of biological weapons by one State Party against another in the course of declared war may thus not be the most common allegation; rather, use in civil war, use by a government against its own citizens, or release in the course of conducting prohibited activity may be more commonly suspected. Thus a State Party as victim is only one of the possible situations that may warrant investigation.
Similarly, the sources providing information about such situations may be quite varied: a State Party; a non-member state; a UN organization; a sub-national group; a non-governmental organization; the news media; or concerned individuals. And the nature of such information may vary considerably, including inter alia: eyewitness accounts by victims, journalists, local health care or scientific professionals, or human rights observers; circumstantial evidence; medical evidence; physical evidence (recovery of alleged delivery devices); or information gathered by national intelligence means and provided to the BWCO by a State Party.
Clearly, the mechanism for initiating investigations needs to be flexible, and because there may be ongoing injury or damage, it should be able to respond rapidly upon receipt of persuasive evidence, from any source, that biological warfare agents have been used or released. Political inhibitions might delay or prevent States Parties from requesting an investigation on behalf of a subnational entity. We therefore propose that the Director-General of the BWCO should have the authority to initiate investigations on his own initiative, as well as at the request of a State Party.
To help the Director-General evaluate available information and determine whether it warrants an investigation, we propose that the executive body of States Parties appoint a Standing Expert Committee on Allegations. Review by the Standing Committee of investigation requests by States Parties could be undertaken at the request of either the Director-General or the executive body; review of proposals for investigation by the Director-General should be mandatory. After the Standing Committee has offered its advice, the subsequent decision whether to proceed or not with the investigation should rest with the requesting party (Director-General or State Party).
Judging the adequacy of epidemiological, medical, and other scientific evidence calls for technical expertise; the Standing Committee should thus be composed of technical experts in the areas of disease, epidemiology, and biological weapons defense. Since speed is essential lest additional evidence be lost or destroyed, the Standing Committee should be prepared to make a recommendation within 24 hours. A Standing Committee of technical experts would obviate the need for specific criteria to eliminate frivolous requests (which criteria would be very difficult to set in advance because of the wide range of types of incident and evidence).
The Director-General should have not only the right, but the duty to propose an investigation when persuasive evidence of use or release of biological weapons agents is obtained. This would provide a degree of automaticity to the response of the BWCO, which would not have to wait for a State Party to make a formal request. The ability of the Director-General to initiate investigations on the basis of evidence provided by, for example, victimized groups within a State Party, would deter such use, and would avoid the need for another State Party to assume the responsibility to speak for the affected group.
Recommendation 3: The Protocol should specify that investigations may be initiated by request of a State Party to the Protocol, or on the initiative of the Director-General of the BWCO, acting on the basis of evidence available to him and with the advice of a Standing Committee of technical experts.
Procedures for Investigating Outbreaks Alleged to be Caused by the Use or Release of Biological Weapons Agents
Procedures for the investigation of incidents in which suspected delivery devices have been recovered or observed would normally include inter alia: (1) a close examination of the devices and of the immediate vicinity (including interviewing possible witnesses of the arrival or discovery of the devices, and the taking of environmental samples for analysis for infectious agents or toxins); (2) the procuring of blood or other samples from persons and animals exposed to the devices to determine possible agent exposure; (3) a survey of human, animal and plant populations for possible effects of agents suspected to have been used; (4) interview of witnesses, persons exposed to the device, personnel at sites that may have been the source of the device, and other relevant persons; and (5) a search over a wide area for other suspect devices.
Investigation of outbreaks of disease suspected to be the result of use or release of biological weapons agents will normally follow standard epidemiological practices to attempt to discover the source of the outbreak. This will involve inter alia: (1) a detailed description of the disease in individuals and its pattern in the affected population; (2) collection of human, animal, plant, and environmental samples (including possible vector species); (3) isolation and identification of the infectious organism or toxin; (4) interviews with affected persons, physicians, veterinarians, public health officials, agronomists and others; (5) charting of the temporal and spatial course of the outbreak to identify its initial focus; and (6) laboratory analysis of pure cultures of infectious agent isolated from samples to identify the individual strain or genetic variant, or of toxin-containing samples to determine the precise chemical structure of the toxin and of any contaminating chemical compounds.
Biological attack or accidental release may be suspected not only in the case of suspicious human disease, but in the case of unusual animal and plant disease outbreaks as well. The process utilized must therefore be flexible and include the necessary expertise to investigate outbreaks in human, animal, and plant populations. Background information on the conduct of investigations of human, animal, or plant outbreaks is given in Appendices II and III, to acquaint non-specialist readers with the elements of typical field investigations.
Each investigation will be unique because of different local circumstances. Thus an effective and flexible investigative mechanism will require the availability of qualified international experts from many disciplines, trained and prepared to participate on very short notice. The composition of the investigating team will be chosen to match the circumstances as they are understood at the time a request for investigation is made. Of course, as the investigation proceeds, new expertise may be needed and the team may thus need to be augmented. Indeed, it may be useful in some circumstances to send a small team initially to assess the situation, and determine if a larger-scale investigation is warranted, and if so what expertise is needed for a full investigation.
Investigating teams should be led by a professional from the BWCO, and should report directly to the Director-General of the BWCO. Team members should be drawn from a list of experts accepted in advance by States Parties, and from among the professional staff of the BWCO and the OPCW. However, provision needs to be made for the occasional use of experts not previously certified, as investigations may need expertise that was not anticipated at the time lists of certified experts were compiled.
The evidence on which the investigative team will base its conclusions will include: (1) various types of samples which are analyzed for suspected agents or for other features that may suggest the presence of an agent; (2) testimony of various people whose experiences or expertise may contribute information useful to the team; and (3) epidemiological and statistical information about the outbreak. The precise nature of the samples and testimony needed will vary according to circumstances.
In many cases crucial evidence is available early but dissipates rapidly. Traces of agent contamination on delivery devices or in the environment may be washed away or degraded (for instance, by UV radiation or by soil microbes); important informants, particularly the earliest infected in an epidemic, may die or disperse; and witnesses' memories become considerably less reliable as time passes. It will be desirable to have the investigating team on site as quickly as possible, within a few days at the most, after a request for investigation is made.
The biggest obstacle to timely investigation of suspicious outbreaks has historically been refusal to allow site access. Full cooperation with investigations initiated under the authority of the Protocol must be recognized as part of the obligations States Parties assume. Cooperation should be seen as including inter alia the obligations to admit the investigating team and their equipment without delay, to help arrange transportation within the country where needed and feasible, to provide security for team personnel and their equipment, and generally to cooperate with the team to facilitate its investigation.
Recommendation 4: The Protocol should include an explicit provision requiring the prompt admission of an investigating team and its equipment by States Parties, provision of safe passage for the team to and from the site of investigation, protection of the team from interference, and cooperation with the investigation.
In some cases the investigative team will require physical access to sensitive sites and personnel. For instance, if the investigation is in response to alleged use in war, the team will require access to the site, to a range of military information on troop disposition and tactics, and to military personnel for interview. Similarly, if the investigation is of an outbreak thought to be a consequence of the accidental release of an agent, the team may need access to industrial, academic, military or other sites which may have been involved. In such cases, site access should follow the guidelines developed for on-site inspections and the protection of confidential information.
The team will need to conduct confidential interviews with involved people (victims of human disease and their relatives, health care workers, public health officials, etc., in the case of human disease; owners of affected animals or crops, animal handlers and agricultural workers, veterinarians, plant pathologists, etc. in the case of plant or animal disease). Confidentiality is necessary to ensure that witnesses or victims are not intimidated and that their testimony does not influence others who will later testify. Interviews need to follow standard protocols, and to be scrupulous in not influencing the interviewee. Interviews should normally be videotaped (or audiotaped if videotaping is unacceptable to the interviewee) for later verification of procedural integrity and accuracy of translation. In many cases, interviews may have to be conducted in a language unfamiliar to the interviewer. In such cases skilled interpretive services will be necessary; these should be provided by the investigative team to insure unbiased translation, although the host country may be requested to assist. A detailed awareness of cultural values and norms different from those of the interviewer is also necessary; when the investigation site lies in an area whose culture is foreign to the investigative team, the team should be supplemented with independent experts on the local culture, who can advise interviewers on the conduct of interviews, the construction of appropriate questions, and the interpretation of answers.
Informed consent may be necessary for obtaining human medical or necropsy samples. The host country should assist in obtaining this. There may, however, occasionally be circumstances in which cultural or religious values prevent the team from gathering certain samples; in such situations, the team should respect the sensitivities. If the team has been adequately briefed, such sensitivities will have been anticipated.
Collection of samples from crop plants or domestic animals may occasionally raise concerns about commercial proprietary information. Such information must be scrupuolously safeguarded.
Samples should be collected in amounts that allow for sample splitting for replicate analysis, and for reananalysis in case of surprising results. The collection of all samples should be recorded and witnessed. When possible, analysis should be done on-site by the team, with appropriate controls. When this is not possible, or when independent confirmation of results is important, samples will have to be transported to approved laboratories for analysis.
The transport to analytical laboratories must be conducted in such a way as to maintain a witnessed chain of custody to guarantee that samples are not tampered with, and that they do not degrade. For biological samples this will often mean that samples are kept refrigerated or frozen until their arrival at analytical laboratories. Packaging needs to be adequate to protect against accidental release of dangerous material in case of accident. Pre-established collecting permits, exit permits, and customs clearances need to be arranged in advance for the exportation and importation of samples.
The length of time necessary to complete an investigation can vary tremendously. In many cases the field component can be completed in a matter of several weeks, with follow-up laboratory work taking an additional few weeks. However, if there are particular complexities associated with an investigation, such as difficult field conditions, a particularly massive outbreak, a novel disease or agent, or the necessity to accumulate background and retrospective information, the investigation may take considerably longer.
If the field work of an investigation has not been completed after one month, the leader of the investigating team should submit to the Director-Gneral a detailed description of the work remaining, and an estimate of the time required to complete it. This should be repeated at monthly intervals if necessary.
When an investigating team arrives on site it must work rapidly and efficiently. Its ability to do so is dependent on many factors, and especially the training and experience of the team. Each team will be composed at least partly of members with previous experience in field epidemiological investigation. Even so, team effectiveness will be greatly enhanced if it has received additional training as a unit. Also, the institutional arrangements, the pre-assembled kits of field equipment, travel arrangements, arrangement for the transport of biological specimens, and all of the other details of such an undertaking will work more effectively after practice. For this reason, we recommend that the procedures be refined and practiced by applying them to natural outbreaks of infectious disease or intoxication indifferent places and circumstances. Thereafter, to maintain readiness, teams should occasionally be made available to international organizations or national governments for use in response to an outbreak on the territory of a State Party (subject, of course, to the approval of the involved State Party).
Recommendation 5: Investigation procedures should be tested and potential team members trained by the BWCO under circumstances as realistic as possible to ensure that they can perform effectively. Periodic exercises should be conducted to maintain readiness.
To perform its functions effectively the BWCO should develop cooperative arrangements with other agencies with related or overlapping interests:
Close cooperation with the Director-General of the Technical Secretariat of the Organization for the Prohibition of Chemical Weapons (OPCW) will be important for several reasons. First, the CWC and the BWC have overlapping coverage of toxin weapons, and they may wish to establish procedures for the joint investigation of alleged use of such weapons. Second, it may not always be obvious at the beginning of an investigation what type of agent is involved (for example, the 1976 outbreak of Legionella in Philadelphia was initially suspected to be due to a toxic chemical). Third, a mixed attack with both chemical and biological agents is possible. And finally, the BWCO and OPCW will maintain lists of qualified professionals and accredited laboratories useful to both organizations.
The BWCO will need to develop and maintain close connections with international organizations that monitor disease outbreaks world-wide, for instance the WHO, the FAO, the OIE, national and regional health organizations, and the Federation of American Scientists' project to promote the establishment of a global Program for Monitoring Emerging Diseases and its electronic conference network, ProMED Mail. Understanding of current patterns of natural disease outbreaks is an important element in the evaluation of possibly suspect ones, and the BWCO will need to be fully aware of the current state of the world's human, animal, and plant diseases. Ongoing investigation of outbreaks by these organizations, or information these organizations receive about national investigations of outbreaks, may also be sources of information useful to the BWCO. These organizations and the BWCO will also have to consider the nature of their overlapping jurisdictions in cases where two or more of them have field teams investigating the same outbreak.
The BWCO should cooperate closely with the Secretary-General of the UN, who is authorized by the UN General Assembly to investigate allegations of violations of the Geneva Protocol, if he calls upon the BWCO for assistance.
States Parties with relevant capabilities should be asked to undertake to maintain on standby, and to contribute for immediate use when needed: logistic support; medical, veterinary and agricultural equipment and materials relevant to investigations under the Protocol; and personnel on the list of qualified experts. Accommodations, guides, transport within the country hosting the investigation, and team security should be provided by the host country if it requested the investigation. All other costs should be borne by the BWCO.
Proposed wording for an annex to a LEGALLY-binding Protocol
Comment: The following proposal for the investigation of allegations of use or release of biological or toxin weapons agents is modeled on the analogous section of the CWC Annex on Implementation and Verification Part XI, "Investigations in Cases of Alleged Use of Chemical Weapons." Significant alterations or additions are noted in italic commentary (deletions of provisions not relevant to the investigation of biological weapons use or release are not noted). More detailed discussion of central provisions is found in the preceding sections of this report. The proposal also assumes that the BWCO will have similar administrative structures with the same titles as the OPCW; these details can be changed as appropriate.
Investigations in Cases of Alleged Use or Release of Biological or Toxin Weapons Agents
A. GENERAL
1. Investigations of alleged use or release of biological or toxin weapons agents shall be conducted in accordance with this document and with detailed procedures to be established by the Director-General of the BWCO. Any proven use of biological weapons agents, or their release during the conduct of prohibited activities, would establish a violation of Article I of the BWC.
Comment: this makes explicit the basis for including investigation of alleged use or release of biological weapons agents in the Protocol.
B. PRE-INVESTIGATION ACTIVITIES
Request for an investigation
2. A request for an investigation of an alleged use or release of biological weapons agents on the territory of any State Party to the Protocol may be submitted to the Director-General by any State Party to the Protocol, together with evidence in support of the allegation.
3. Alternatively, the Director-General shall propose an investigation on his own initiative if he is in possession of information that suggests that biological or toxin weapons agents may have been or released on the territory of a State Party.
Comment: this provides for the investigation of suspect outbreaks on the initiative of the Director-General of the BWCO, in addition to investigations requested by a State Party. Information available to the Director-General may lead him to conclude that an investigation is warranted even if one has not been requested by a State Party.
4. To the extent possible, requests by States Parties or proposals by the Director-General for investigation should include the following information:
(a) The party requesting the investigation;
(b) The State Party on whose territory use or release of biological weapons agents is alleged to have taken place;
Comment: in many cases an affected State Party affected will request an investigation on its own territory. However, it is necessary to provide for investigations on the territory of other States Parties, for instance in case of suspected accidental release, or in the case of the use of biological or toxin weapons agents in civil conflict.
(c) The point of entry or other suggested safe routes of access;
(d) Location and characteristics of the areas where biological weapons agents are alleged to have been used or released;
(e) When use or release of biological weapons agents is alleged to have occurred;
(f) Types of delivery systems believed to have been used;
(g) Nature and extent of alleged use or release;
(h) Agents whose use or release is suspected, and a history of their prior occurrence in the region;
(i) Effects on humans, animals, and plants;
(j) Basic epidemiological information on any spread of disease from the point of alleged use or release;
Comment: because biological agents can initiate epidemics that can spread from their point of origin, and may continually generate new casualties over an extended period, it is desirable for requesting Parties to describe to the extent possible the time course, geographic extent, and demographic characteristics of any epidemic spread, along with any available information on items such as incubation time, duration of illness, case fatality rate, and other epidemiological features.
(k) Request for specific assistance to control an outbreak, if needed.
Comment: the investigating team should, to the extent possible and as need dictates, assist in the control of the outbreak while carrying out its investigative work. The Director-General could also ask WHO, OIE, FAO, or other appropriate groups for assistance in controlling the outbreak if they are not already participating.
5. The party that has requested an investigation may submit at any time any additional information it deems necessary. If additional information is requested by the Director-General of a requesting State Party, it should be provided within 24 hours, or reasons given why it is unavailable, and when it will become available.
Consultation with a Standing Committee of Technical Experts
6. A Standing Expert Committee on Allegations shall be appointed by the Executive Council from a slate of experts nominated by States Parties. The Committee, composed of technical experts in the areas of disease, toxicology, epidemiology and biological weapons, shall provide advice, within 24 hours, on the evaluation of evidence submitted in support of a request for an investigation. No Committee members who are nationals of States Parties directly involved shall take part in the consideration of a particular request.
7. Before proceeding with an investigation on his own initiative, the Director-General shall consult the Standing Committee. The Standing Committee shall provide its advice within 24 hours. In the light of this advice the Director-General shall then decide whether or not to proceed with preparations for the dispatch of an investigation team.
8. Immediately upon receipt of a request by a State Party for an investigation, the Director-General shall proceed with preparations for the dispatch of an investigation team. The Director-General may then consult the Standing Committee at his discretion or at the request of the Executive Council. The Standing Committee shall, within 24 hours, provide its advice to the Director-General and to the requesting State Party, which shall then notify the Director-General whether or not it wishes him to proceed with the investigation.
Comment: The Standing Committee would provide expert technical advice on the significance of the evidence submitted. Appointment of the Standing Committee by the Executive Council insulates it from the Director-General. In case of requests for an investigation originating with the Director-General, review by the Standing Committee should be mandatory; for requests by States Parties, the Standing Committee could be consulted if the Director-General or the Executive Council has doubts about the evidence presented. In either case, after receiving the Standing Committee's advice, the decision whether to proceed would be made by the original requesting party
Notifications
9. The Director-General shall immediately acknowledge receipt of a request from a State Party and inform the Executive Council and all States Parties.
10. The Director shall immediately inform the Executive Council and all States Parties of all recommendations of the Standing Expert Committee on Allegations, and of every decision regarding the dispatch of an investigating team.
11. The Director-General shall immediately notify the State Party on whose territory an investigation is to be conducted. The Director-General shall also notify other States Parties if access to their territories might be required during the investigation.
Assignment of investigation team
12. Immediately upon receipt of a request by a State Party, or upon proposing an investigation on his own initiative, the Director-General shall select the inspection team, taking into account the circumstances and specific nature of a particular request, and alert it for possible dispatch within 24 hours.
13. The Director-General shall prepare a list of qualified and trained experts whose particular field of expertise could be required in an investigation of alleged use or release of biological weapons agents, and constantly keep this list updated. This list shall be communicated to States Parties, in writing, within thirty days after entry into force of the Protocol and after each change to the list. Any qualified expert included in this list shall be regarded as designated, and shall be eligible for any investigation, unless a State Party, not later than 30 days after its receipt of the list, declares its non-acceptance in writing. In case of non-acceptance, that expert shall not participate in investigations involving the State Party that declared its non-acceptance.
14. One or more core teams, with alternates and potential ad hoc members, shall be trained in advance and provided with immunizations and necessary papers so that they can be called upon for assignment at short notice. The leader of each team shall be drawn from the BWCO inspectorate; other team members shall be selected from the BWCO inspectorate, from the list of qualified experts maintained under paragraph 11 above, from the list of experts maintained by the Director-General of the OPCW, or from inspectors and assistants designated for challenge inspections by the OPCW.
Comment: Effective immunization may take weeks or months, but teams must be prepared to go into the field with as little as 24 hours notice. Previous training as a team under field conditions similar to those likely to be encountered in a real investigation is important: teams should be made available to international health organizations and national governments for occasional use in response to outbreaks on the territory of States Parties.
Because close cooperation between the BWCO and the OPCW is expected, and because OPCW inspectors and designated experts may well have expertise useful to a BW investigation, it will be efficient to utilize them as well as inspectors and designated experts associated with the BWCO.
15. No nationals of a requesting State Party, the host State Party (on whose territory the investigation takes place), or an accused aggressor State shall be a member of the investigation team.
Comment: this provision is drawn in part from the CWC Verification Annex Part X(2) relating to the conduct of challenge inspections.
16. If in the judgement of the Director-General the investigation team requires expertise not represented on the list of approved experts, or among the approved inspectors and assistants of the BWCO or the OPCW, he may propose to the Executive Council that the team be augmented by the addition of specified experts.
Comment: the expertise needed in an investigation of alleged use or release cannot always be anticipated. In many cases, organism-specific or place-specific expertise not anticipated at the time of compiling the list of experts may be of great importance. Some provision for calling upon experts not holding prior approval is necessary.
17. The host State Party shall have the right to attach an observer to the investigation team. The observer shall have the right to observe all activities of the investigation team, except interviews and medical examinations, unless the team leader determines that such observation would interfere with the team's work.
Comment: this provision is drawn in part from the Report of the Secretary-General of the UN on Chemical and Bacteriological (Biological) Weapons. The exception for interviews and medical examinations reflects the danger of intimidation of interviewees if they are unsure of the confidentiality of their testimony, and the necessary protection of the right to privacy of medical patients.
18. When briefing the investigation team, the Director-General shall include any additional information provided by a requesting State Party, or any other sources, to insure that the investigation can be carried out in the most effective and expedient manner.
Dispatch of investigation team
19. As soon as an investigation team has been selected, the Director-General shall, through contacts with the relevant States Parties, request and confirm arrangements for the safe reception of the team.
20. The Director-General shall dispatch the team at the earliest opportunity, taking into account the safety of the team.
21. If the investigation team has not been dispatched within 24 hours from the receipt of the request, the Director-General shall inform the Executive Council and the States Parties concerned about the reasons for the delay, and he shall continue to do so at intervals of 24 hours until the team is dispatched or the investigation is cancelled.
Briefings
22. The investigation team shall have the right to be briefed by representatives of the host State Party upon arrival and at any time during the investigation.
23. Before the commencement of the investigation the investigation team shall prepare an investigation plan to serve, inter alia, as a basis for logistic and safety arrangements. The investigation plan shall be updated as need arises.
C. CONDUCT OF INVESTIGATIONS
Access
24. The investigation team shall have the right of access to any and all areas which may be affected by the alleged use or release of biological weapons agents, including all areas to which an outbreak under investigation has spread. It shall also have the right of access to any location or facility that may have been the source of the alleged use or release, hospitals, refugee camps and other locations it deems relevant to the effective investigation of the alleged use or release of biological weapons agents. For such access, the investigation team shall consult with the host State Party.
Comment: this wording gives the investigation team the right of access to all of the territory affected by an outbreak, and to lacations that may be suspected as its source. Such access is required for thorough epidemiological study of outbreaks caused by infectious agents, unlike those caused by chemical and toxin weapons, which do not have the potential for epidemic spread.
Sampling
25. The investigation team shall have the right to collect samples of types, and in quantities it considers necessary. If the investigation team deems it necessary, and if so requested by it, the host State Party shall assist in the collection of samples under the supervision of members of the investigation team. The host State Party shall also permit and cooperate in the collection of appropriate control samples from areas neighboring the site of the alleged use or release and from other areas as requested by the investigation team.
26. Samples of importance in the investigation of alleged use include biological toxins, infectious agents, delivery devices or remnants thereof, environmental samples (air, soil, vegetation, water, snow, arthropods, rodents and other wild animals as possible reservoirs and vectors, etc.), biomedical samples from human or animal sources (blood, urine, excreta, tissue, etc.), and tissue from affected and control plants. The collection of human samples will require informed consent, which the inspected State Party shall assist in obtaining.
Comment: the list of examples of sample types reflects the greater breadth of sampling likely to be required by a biological investigation. Note that human samples require informed consent; the inspected State Party should cooperate with the investigation team to obtain such consent, or to devise alternative means of obtaining information if cultural practices or religious beliefs prevent the team from obtaining samples.
27. The Director-General shall, upon nomination by States Parties, test and accredit reference laboratories to be certified to perform analysis of samples when called upon. A list of certified reference laboratories shall be communicated to States Parties, in writing, within ninety days after entry into force of the Protocol and after each change to the list.
Comment: this provision recognizes the need for a network of independent laboratories in different contries capable of performing the microbiological and toxicological analysis that will be required to confirm the field results of investigating teams, and to perform analytic tasks too sophisticated to be performed in the field.
28. If requested by the investigation team, the inspected State Party shall provide access to all biological samples, and to microbial cultures derived therefrom, taken at any time before the alleged use or release, to serve as reference or control samples.
Comment: this provision, not part of the CWC, is necessary to allow laboratory analysis to determine if the particular strain of biological agent, or the toxic compound, has been naturally present in the area previously.
Extension of investigation site
29. If the investigation team during an investigation deems it necessary to extend the investigation into a neighboring State Party, the Director-General shall notify that State Party about the need for access to its territory and request and confirm arrangements for the safe reception of the team.
Extension of investigation duration
30. If the field work of an investigation has not been completed after one month from the date of dispatch of the team, the leader of the team shall submit to the Director-General a request for extension of the duration of the investigation, accompanied by a detailed description of work remaining to be done, and a timetable for completion of the work. Similar statement shall be submitted, if necessary, at monthly intervals.
31. If the investigation team deems that safe access to a specific area relevant to the investigation is not possible, the Director-General shall be informed immediately, and he will arrange with the host State Party for necessary security measures. If necessary, the period of investigation shall be extended until safe access can be provided and the investigation team will have concluded its mission.
Expansion of investigation team
32. If the investigation team deems that it lacks specific expertise necessary to the conduct of the investigation, it shall immediately so inform the Director-General, who will augment the team as appropriate from his list of experts, from the list maintained by the Director-General OPCW, or from inspectors and investigation assistants designated for challenge inspections by the BWCO or the OPCW. If no experts with the needed expertise are certified by the BWCO or the OPCW, the Director-General may, with the approval of the Executive Council, appoint an independent expert to the team.
Comment: this provision, not present in the CWC, reflects the complexity of epidemiological investigation, which can easily lead in unexpected directions and require unanticipated expertise.
Interviews
33. The investigation team shall have the right to interview and examine persons, and to examine animals and plants, which may have been affected by the alleged use or release of biological weapons agents, including those affected by spread of the outbreak from its initial locus. It shall also have the right to interview witnesses of the alleged use or release, and any others who might have information about the source or nature of the outbreak, including medical personnel, public health officials, veterinarians, agronomists, and other persons who have treated or come into contact with persons, plants, or animals that may have been affected by the alleged use or release, or any others who may have relevant information. The investigation team shall have access to medical records if available, and shall be permitted to participate in medical examinations and autopsies, as appropriate, of persons or animals that may have been affected by the alleged use or release of biological or toxin weapons agents.
Comment: this provision expands slightly the authority of investigation teams, for instance by giving them the right to participate in medical examinations as well as autopsies. Such participation may be critical in identifying the infectious agent or toxin and in deducing its mode of transmission.
D. REPORTS
Procedures
34. The investigation team shall, not later than 72 hours after its arrival on the territory of the inspected State Party, send a situation report to the Director-General. It shall further throughout the investigation send progress reports as necessary.
35. The investigation team shall, not later than 72 hours after its return from the investigation site, submit a preliminary report to the Director-General. A final report shall be submitted to the Director-General not more than 30 days later. The Director-General shall promptly transmit the final report to the Executive Council and to all States Parties. In drafting the final report, the investigating team may, with the approval of the Director-General, request the assistance of additional experts as necessary.
Comment: this provision differs in two substantive ways from the analogous paragraph in the CWC: it allows the preliminary report to be held in confidence by the Director-General; and it provides for the involvement of additional experts in the drafting stage of the report. The former provision recognizes that the preliminary report, lacking as it will a full analysis of all data, is not ready for release to States Parties. The latter provision recognizes that the expertise necessary for data analysis may differ from that required for field investigation.
36. If laboratory work is ongoing at the time of the submission of the final report, the report shall note that further information will be submitted, estimate the time at which such information will be available, and note its significance. When such work is completed, its results shall be reported to the Director-General in a supplementary report, including any revisions of the final report that may be necessary.
Comment: this provision recognizes that some laboratory studies and background research may require more time that is allowed by the timetable for the production of the final report.
Contents of the Reports p 37. The situation report shall indicate any urgent need for technical, medical, veterinary, or agronomic assistance and any other relevant information. The progress reports shall indicate any further need for assistance that might be identified during the course of the investigation.
38. The final report shall summarize the factual findings of the investigation, particularly with regard to the alleged use or release cited in the request. In addition, the report shall include a description of the investigation process, tracing its various stages, with special reference to:
(a) The locations and times of sampling and on-site analysis.
(b) Supporting evidence, such as the records of interviews, the results of medical examinations and epidemiological and scientific analyses, and the documents examined by the investigation team.
(c) An account of the assistance, and its timeliness, provided by the host State Party.
39. If the investigation team collects any information in the course of its investigation that might serve to identify the origin of any biological agent or toxin found at the site, that information shall be included in the report. Such evidence may include, inter alia, chemical composition and the presence of inert materials in the case of possible toxin weapons, and serological or molecular sequence evidence in the case of infectious agents. In either case, the report shall present such environmental and historical information as is available on the previous presence of the alleged agent in the region.
Comment: The finding of a chemical agent is often prima facie evidence of weapon use. In the case of infectious agents, presence alone is rarely definitive. Thus evidence of geographic origin, and analysis of prior presence or absence of the agent in the region, may be important factors in determining whether there was in fact weapon use or release. Detailed laboratory analysis of the molecular characteristics of the agent, and comparison with those of agents isolated at other times and places, may help to identify the geographic origin of the agent. Detailed interviews with public health professionals, veterinarians, agronomists, and government officials with responsibility for monitoring diseases can help develop a profile of previous presence of the agent in the area, and this can sometimes be verified by analysis of stored biological or environmental samples from before the outbreak, or from unaffected nearby areas.
E. STATES NOT PARTY TO THE BWC AND THE PROTOCOL
40. In the case of alleged use or release of biological weapons involving a State not Party to the BWC and the Protocol, the BWCO shall cooperate with the Secretary-General of the United Nations. If so requested, the BWCO shall put its resources at the disposal of the Secretary-General of the United Nations.
DESCRIPTION OF PROCEDURES COMMONLY USED IN THE FIELD INVESTIGATION OF A SUSPECT OUTBREAK OF INFECTIOUS DISEASE OR INTOXICATION OF HUMANS AND ANIMALS
Introduction
We describe here very briefly the conduct of an epidemiological field investigation of human or animal disease, as applied to a suspicious outbreak that might be investigated by the BWCO. Obviously the details may vary considerably according to whether an infectious disease or intoxication is under investigation; whether humans, animal or both are involved; whether a vector is involved; whether disease is caused by a known organism or compound, or is novel; where the outbreak is located; the season and weather; whether the investigation occurs in the context of peacetime, civil unrest, or overt war; etc. However, much of the work of an investigating team will consist of common procedures and approaches, as summarized here. Indeed, in many cases investigations may be considerably less complex than we describe here, for instance if the etiologic agent is identified in advance and the scope of the outbreak is limited. Note that this account is not intended to present guidelines, but rather to describe conduct typical of field investigations to acquaint non-expert readers with some elements of the process.
A typical general-purpose team might include a physician with expertise in infectious disease, a pathologist, a microbiologist, a toxicologist, a veterinary clinician, a veterinary pathologist, and an epidemiologist. If the etiologic agent is known or strongly suspected, an expert on it would be included. All team members would be trained in epidemiological field investigation. This general-purpose team, constituted to investigate outbreaks of zoonotic disease, could be supplemented or otherwise modified as appropriate. Interpreters, security personnel, communications experts, anthropologists, and other support personnel would accompany the team as needed.
Typically the team will set up headquarters in a site convenient to the site of the outbreak, that will allow its members ready access to victims of the outbreak, and to international transportation if possible. Here, samples can be sorted and catalogued, blood samples centrifuged and serum drawn off, aliquots prepared, tissues put in fresh fixative, and all samples packed and stored appropriately. The team's field laboratory will be located here, and many routine serological, microbiological, and other diagnostic tests will be performed immediately. Daily records of interviews, clinical findings, and laboratory results are entered into computers, and back-up copies made. Headquarters also provides a place for team members not in the field to meet daily for discussion and planning.
Particularly in the case of animal outbreaks, or in the case of widespread human outbreaks, part of the team may be travelling in the field for many days at a time. In such cases, a mobile sample processing unit is required, that can be set up at a temporary location, and quickly disassembled when the team moves to a new location. Reliable and trustworthy courier service between field sites and headquarters will often be needed.
It is also desirable for the headquarters to be convenient to local diagnostic laboratory facilities, with capabilities in such areas as hematology, biochemistry, serology, microbiology, radiology, etc., since the team's work may be aided substantially by permanent facilities to supplement their own field laboratory.
Interviews with responsible personnel from the host country, such as regional or local public health officials, hospital physicians, and veterinarians may be helpful in assessing whether the outbreak is consistent with disease patterns in the area. Such local contacts may also be helpful in generating a tentative case definition, to allow the team to determine which cases are part of the outbreak and which are due to coexisting disease. These case definitions are typically quite broad at the beginning of an investigation, to prevent relevant cases from being overlooked, but they are usually made more restrictive as the identity of the disease becomes clear.
Interviews with responsible local officials will attempt to define the approximate date and location for the beginning of the outbreak. If this is not obvious, it will be reconstructed by plotting on a map the location and time of onset of cases that fit the case definition, and by charting on graph paper the incidence of new cases as a function of time. Such geographic and temporal plotting can also reveal attributes of an outbreak that might suggest deliberate use; for instance, the simultaneous onset of pneumonic plague or pulmonary anthrax in large numbers of victims might suggest exposure to an unnatural aerosol. Interviews with patients (or farmers in the case of animal outbreaks) will attempt to define common exposures that might suggest the source of contact with the agent.
Identification of the disease or toxin involved will generally involve a combination of approaches. The first step will be a definition of the clinical features of the disease, including clinical symptoms and signs, laboratory results, X-ray results, and macroscopic and microscopic pathology. In some cases the clinical findings may be nearly definitive; in others they may be consistent with a wide variety of etiologies. Where an infectious agent is suspected, the investigation will include microscopic examination and microbiological cultures of body fluids and/or tissues, together with serological tests for changes in antibody levels (preferably with paired acute and convalescent sera). Where there have been fatal cases, post-mortem macroscopic and microscopic pathological findings may be helpful in identifying the agent. When intoxication is suspected, chemical and toxicological tests on blood, urine, stool, hair, biopsy or necropsy tissue, or other material are performed. In many cases these tests may be quite simple, within the scope of the team's field laboratory or of local diagnostic and microbiological laboratories. In other cases they will require a level of sophistication that may not be available in the field or in local facilities; in such cases, samples will need to be transported to distant laboratory facilities, and the results communicated promptly to the team.
Medical examinations will normally conducted in local clinical facilities or in the field; autopsies are generally performed in local hospitals. Veterinary examinations and necropsies are normally conducted in the field. Veterinary teams consequently need to be equipped to disinfect necropsy sites, and need to coordinate carcass disposal with local officials. Burial is often inadequate, as scavengers can exhume carcasses unless they are deeply buried. Incineration with "domestic napalm" (soap powder and gasoline) is effective where other options are not available.
Field teams, whether seeking human or veterinary cases, need to be equipped with accurate maps, satellite imagery, GPS receivers, and meterological information from the time of the beginning of the outbreak.
In addition to samples from affected humans and animals, it may be necessary for the team to sample apparently unaffected people, livestock, wild animals, plants, soil, and water as controls, and to determine the geographic limits of affected habitats. Stored biomedical samples from before the outbreak may also be tested for presence of the agent, or for other features that might indicate its previous presence. Similarly, if a toxin is identified, environmental samples are taken to determine if the toxin is naturally present.
Once the etiologic agent of an infectious disease has been identified, its more detailed characterization will require study with any of a variety of molecular approaches. Typically a region of its genome will be sequenced, or the distribution of restriction sites in its genome will be determined by using restriction endonucleases. Such techniques, performed in approved reference laboratories, will provide a level of detailed knowledge of the etiologic agent that may allow its precise geographic origin to be inferred.
Necessary Access
To perform its investigation most effectively, the investigating team will need the following access, inter alia: to medical and veterinary records in the area of the outbreak; to human and/or animal victims of the outbreak (including currently ill, convalescent, and post-convalescent humans and/or animals) for examination, for interviews, and for the taking of biological samples; to deceased victims of the outbreak for examination and sampling; to exposed but unaffected human and animal populations, and to unexposed human and animal populations, for sampling to determine the level of subclinical disease and of previous exposure; to environmental samples in the area of the outbreak and elsewhere as controls and for the detection of vector or reservoir organisms; and to biomedical, veterinary, or environmental samples taken by the host country prior to the arrival of the team (including ones taken before the outbreak). Since many diseases can affect more than one species (for instance, many human diseases affect animals as well), teams will often need to proceed with human and veterinary investigations in a parallel and coordinated fashion.
The team will not normally require access to sensitive sites, such as military installations, battlefields, prisons, or proprietary industrial or agricultural sites. However, in some cases the geographic distribution of cases, or the search for devices or other sources of infection, may lead to a need for access to such sites. Such access would also likely be required in investigations of alleged release of a biological weapons agent.
Sample Collection and Handling
Sample collection from humans requires informed consent. Samples are routinely coded so that investigators can identify their source in case additional sampling or interview is necessary, but confidentiality is otherwise scrupulously maintained. Samples taken in the field are recorded such that the exact place can be resampled if necessary. The team should be equipped with GPS receivers for this purpose.
Critical samples are taken in amounts sufficient for subdivision for replicate analysis in different laboratories. Samples are treated to reduce post-sampling degradation to a minimum: refrigeration is routine, and freezing is common; preservatives may be used as well. Such caution is necessary even if a particular robust infectious agent or stable toxin is suspected, as it may be necessary to later analyze samples for other, initially unsuspected agents. Samples are normally sealed, and they are treated with the caution appropriate to a sample that may contain an infectious or toxic agent. In the case of an investigation by the BWCO, samples would additionally have to be scrupulously protected from tampering. Thus their seals and labels would have to be tamper-proof, and they should be continuously in the possession of responsible officials from the time they are collected to the time their analysis is completed.
Samples are coded so that laboratory analysis is blind. Some samples should be supplemented with known amounts of agent as a test of the accuracy of laboratory analysis.
In order to avoid delays in transporting samples to specialized laboratories in countries other than the host country, advance clearance should be obtained from appropriate officials to avoid quarantine or other delays. Such clearance should be arranged before departure of the team, or immediately when the need arrives, and should be the responsibility of the BWCO. The team should verify clearances if they pass through involved international airports en route.
Since appropriate samples are usually critical to an effective investigation, every effort should be made to secure them. However, in some cases religious beliefs or cultural patterns may make this difficult. The investigating team must be respectful of such constraints, but should make strenuous efforts to alleviate them where possible. This may require the team to possess anthropological expertise, and to enlist the support of local religious and cultural leaders in the course of their investigation.
Conduct of Interviews
Interviews are conducted in a language in which both team member and interviewee are fluent. If this is not possible, expert translation is necessary. Since obtaining accurate answers may require a considerable level of cultural sensitivity as well as linguistic fluency, investigating teamsshould have familiarity with the culture in which they are working. If they do not, the team should be supplemented to ensure it. Interviews should be conducted by team members experienced in cross-cultural interviewing, such as medical practitioners or behavioral scientists.
Where abundant time or small numbers of victims allow, it may be feasible to interview and/or examine all victims or all affected farmers. Otherwise, the selection of victims and farmers for interview should made by the team, with the intent to sample randomly.
Interviews are normally conducted based on a standard questionnaire used by all interviewers, although there will be flexibility for individual interviewers to follow up leads as they arise, and for interviewees to volunteer information not solicited. These forms are tailored to the investigation, and are changed as needed to reflect the changing nature of the investigation. Interviews follow standard social science interview procedures, and are framed carefully to protect against leading the interviewee. To protect against potential intimidation of interviewees, interviewers should insist on privacy during the interview. All interviews should be videotaped or audiotaped for later retrospective analysis, unless doing so violates cultural or religious beliefs, or is unusually intimidating to interviewees.
Questionnaires should be carefully prepared with a view to rapid data entry and analysis either in the field or immediately upon return. Record-keeping in the field is normally on portable computers; multiple back-up copies are maintained of all files.
Necessary Field Equipment
Before arriving on site, the investigating team may have an idea of the identity of the disease they will find (it will often have been diagnosed with certainty by the host country). Even if not, the range of possibilities will often be fairly limited. This allows the team to tailor its kit of reagents and equipment appropriately.
The team will normally be equipped with sufficient field laboratory material to allow it to perform needed diagnostic tests and to culture suspected etiologic agents. Portable refrigerators and freezers are used to preserve reagents and cultures, and for temporary storage of samples. Reagents and equipment are crated to allow easy transportation, ready access, and to protect them from damage during transit. Back-up power supplies for computers, refrigerators and other critical items are included.
Team personnel will have access to field biosafety equipment, and will be equipped to decontaminate possible contaminated material. Provision for secure and safe shipment of samples at an appropriate containment level should be arranged in advance.
The team needs a secure form of communications, both locally to communicate with its members and with local officials, and globally to communicate with the BWCO and with other experts. Voice, fax, email, and data transmission capabilities are desirable. GPS units are available for precise determination of the location of any sites of importance. A full complement of maps, satellite imagery, and meteriological information in both hard copy (for posting at headquarters) and in electronic form (for portability in the field) is also needed.
Necessary Diagnostic Laboratories
In many cases, the investigating team will be able to conduct necessary laboratory work with its own field facilities. In other cases, local laboratories may cooperate in the investigation. In the latter case, it is necessary for a team member to carefully oversee all operations to prevent tampering, and to insure the provenance of all samples and the integrity of all reagents. Freezer packs and coolers for sample preservation during transport must be part of the team's equipment. However, even when field or local laboratory facilities are adequate to perform needed analysis, critical findings will need to be confirmed by laboratories certified by the Director-General.
The field team also needs rapid access to back-up laboratories that can perform sophisticated molecular tests, electron microscopy, toxicology, analytical chemistry, etc., and that can culture a range of potential agents encompassing bacteria, protozoa, fungi, and viruses. Given the range of expertise necessary, a network of existing laboratories would be established and certified. At least some laboratories need to have the capability of handling highly dangerous organisms; in some cases, BL 4 facilities may be necessary.
Introduction
We describe here very briefly the typical conduct of an epidemiological field investigation of plant disease, as applied to a suspicious outbreak investigated by the BWCO. Obviously the details may vary considerably according to the nature of the infectious agent; whether animal vectors are involved; whether the disease is caused by a known organism, or is novel; where the outbreak is located; the season and weather; whether the investigation occurs in the context of peacetime, civil unrest, or overt war; etc. However, much of the work of an investigating team will consist of common procedures and approaches, as summarized here. Note that this is not intended to present guidelines, but rather to describe conduct typical of field investigations to acquaint non-expert readers with some elements of the process.
Fungal pathogens are notorious for their ability, given appropriate environmental conditions, to spread rapidly. Fungi and viruses cause the greatest yield losses, although epidemics of bacteria and phytoplasmas can also be devastating. Fungal and bacterial diseases, much like insect pests, invade and build up in a target field, causing damage in later stages of plant development. If they are observed in time, curative measures can often be taken to minimize damage. In contrast, infections by plant viruses are usually systemic; by the time symptoms indicate that disease is present, remedial actions are worthlesse. Moreover, secondary spread may have already occurred before symptoms are noted. Viral diseases are difficult to prevent and not possible to "cure" once infection has occurred; crops may be rapidly invaded and totally lost.
Intentional plant outbreaks could be induced by the introduction, into the target region of new organisms or more damaging forms of existing organisms. In the case of plant viruses, approximately 90% of the 800 viruses currently recognized are transmitted by biological vectors, principally insects. Thus, outbreaks of plant viral disease could be effected by the introduction of infected vectors, or of more efficient vectors. Delivery systems for plant pests might include: a) propagative material, i.e. seeds or reproductive plant parts (for seed-borne pathogens or pathogens in vegetatively propagated crops); b) biological vectors (for plant viruses): c) contaminated materials such as soil (for soil-borne pathogens or weed seed), water (for water-borne viruses) or inert materials (for stable, highly contagious plant pathogens); and d) physical containers (for fungal spores or invertebrate eggs, larvae or adults).
Teams should include expertise in plant virology, botanical epidemiology (especially plant virus epidemiology), plant pathology, vector and pest entomology, and agrochemical control. The specialists should have a crop-based expertise as opposed to taxonomic expertise, i.e. a citrus virus expert rather than a potyvirus expert.
Preparation includes gathering of field equipment (including computer equipment for communication and in-field analysis of epidemiological data); organization of communication with headquarters and back-up support from colleagues; collection and review of all reference literature available on the pests, plant protection practices and ecology/environment of the region under alleged attack; and briefing on the political and socio-economic structure of the country and regions to be visited. Many of these activities can be completed in advance as part of the stand-by preparation f or investigation.
Necessary Access
It is necessary to work in the specific fields where the alleged introduction has taken place, as well as in the general region. Thus, it is necessary to acquire direct physical access to the plant communities (production fields, orchards, forests, etc.) that are infected as well as uninfected fields in the vicinity. It is also necessary to have access to individuals that must be interviewed (e.g. producers, field technical and extension personal, national professionals, government-ministry personal, etc.), and the national literature and data bases needed for the study (e.g. meteorological data, topographic maps, land-use data, etc.)
Sample Collection and Handling
Pathogen collection
Fungi and bacteria. Leaves, pods, stems or other tissue infected with fungus or bacteria are collected. Plant tissue should be young, green and fresh (not senescent). Collected tissue is stored in absorbent paper toweling placed in vented containers to avoid moisture build-up that could cause growth of saprophytic fungi and destroy samples. After initial drying, tissue is placed in airtight vials with a drying agent (silica gel, calcium chloride).
Virus, viroids and phytoplasmas. For leaf tissue from annual plants, both young and fully expanded leaves are collected from plants with initial and advanced symptoms. Leaf tissue should be dried in airtight vials containing a drying agent (silica gel, calcium chloride). Leaf material with advanced symptoms should be placed in vials containing a fixative (e.g. gluteraldehye). This tissue can later be processed for electron microscopy (cytopathology).
For collection of tissue from perennial plants, the ideal tissue is bark peeled from the stem of tender new growth at a point where the leaves are nearly fully expanded and not yet mature. Shoots from several locations around the tree are sampled. If no new growth is available, immature fruit may be collected.
Root material. Root material from infected plants, as well as soil collected from around the roots of infected plants, should be collected, placed in containers, and stored in an ice chest for subsequent processing.
Collection of seed. If still available, samples of the seeds or planting material used by producers in the infected fields, should be collected, and kept cool and dry, for subsequent pathogen analysis.
Invertebrate collection
Invertebrates may be vectors of plant pathogens. The most common in this role are aphids, whiteflies, leafhoppers, planthoppers, beetles and thrips.
Aphids are normally collected directly from the host plant, using a camel hair's brush, and placed directly into alcohol. A number of adults may be necessary for accurate identification, and where there is a choice both alate (winged) and apterous (wingless) adults should be taken. If only apterous aphids are present (and time permits), collect a leaf or twig and place in a glass bottle, covered with netting. Often winged adults will emerge, and those can then be collected into alcohol for identification.
Whitefly puparia are usually best collected whilst still attached to leaf tissue. Either the whole leaf may be excised and stored dry until required for preparation, or pieces of leaf with puparia attached may be placed and stored in alcohol prior to treatment. An advantage of dry storage is that adults may emerge inside the envelope and this be reliably correlated with the pupal cases - taxonomic keys based on adult characters are becoming available.
Leafhoppers, planthoppers and beetles should be collected using sweep nets, D-vacs, or aspirators and placed into killing jars. These insects should be pinned within 24 hours to avoid desiccation. If that is not possible, then insects should be preserved in alcohol. Thrips are collected with a sweep net and stored in preservative.
Invertebrate pests. Collection and preservation methods vary greatly. Organisms should be identified to Order, in the field, so that the appropriate collection and preservation techniques are employed. Collection of adult specimen is often necessary in order to obtain complete identifications of the organism. Collection methods include sweep and aerial netting, beating, trapping and aspiration. In general, non-insect invertebrates, soft-bodied insects and immature insects are collected into fluid, usually alcohol, to avoid desiccation. Hard bodied, adult insects are most often dry-mounted on entomological pins. Specific methodologies for collecting and preserving invertebrates are readily available in the technical literature.
Plant collection
It may be desirable to collect target plants that display characteristic symptoms or injury, or non-target plants that are hosts for suspect organisms. In such cases, multiple plant specimen are collected. If available, specimen that have flowers and/or fruits are collected, since these are often necessary for proper identification. Plant samples are dried in a plant press.
Population samples
In addition to samples of individual organisms, for the purpose of identification and genetic typing, suspect pest populations should also be sampled within the affected field/area and in nearby control areas, for the purpose of analyzing population structure. Natural populations exhibit a heterogeneous structure, whereas introduced populations would tend to be relatively homogeneous in their structure.
Conduct of Interviews
Ideally, symptoms of infection or plant injury should be described and collected by the team members, and pictures or video footage should be taken. However, if affected plants are no longer present, then the producer/manager should be asked to describe the symptoms. Additional descriptive information to be collected should include: plant variety, source of crop seed or planting material, cropping/management practices (especially use of agrochemicals), history of field (i.e. crops planted previously, and previous use of agrochemicals), planting and germination dates, and any information on weather conditions (rainfall, drought, wind patterns). If a retrospective study, then any yield data should be obtained. Identical information should be collected for non-affected fields in the area.
Interviewees should be asked when affected plants were first noticed, where in the field symptoms were first noticed, how the outbreak spread (in time and space), and the actual spatial pattern of outbreak within the field, if possible. If the outbreak is on-going, the team members should also estimate incidence and severity. Perennial crop plants should be inspected for unusual physical damage that could serve as an entryway for non-vectored pathogens.
It should be ascertained whether producers/managers observed populations of insects (potential vectors, that may no longer be present) in unusually high numbers or exhibiting abnormal behavior (e.g. unusually early or simultaneous immigration).
Interviews of the producer/managers should include their knowledge on cropping patterns and conditions in the region, i.e. what their neighbors are planting (especially upwind of their fields); if there have been any noticeable changes in cropping practices, cropping patterns or land use in the last several years; whether weather conditions were normal or abnormal; average yields for the region and for their production units.
Producers should also be questioned about unusual movement of people, vehicles or aircraft not previously observed in the area. And, the infected fields and surrounding areas should be searched for possible containers (e.g. petri plates, vials) that could have been used as part of a delivery system.
Precise latitudes, longitudes and altitudes should be recorded with a GPS unit for all field sites visited.
Field Equipment and Materials
Field equipment will be similar to that needed for human or animal outbreaks, except that team biosafety is not normally such an issue. Specialized collecting equipment (nets, aspirators, etc) is included.
Diagnostic Laboratories
In Europe and North America, research institutions are highly specialized, and often research units work on a restricted set of (pathogen or invertebrate) taxa. However, collectively, these institutions are capable of carrying out identification and analysis of most plant pests in temperate plant communities of importance.
Although few developing-economy countries possess comparable laboratory facilities, the regional International Agricultural Research Centers (IARCs) of the Consultative Group on International Agricultural Research (CGIAR) could also be utilized as reference laboratories for investigations. The CGIAR is jointly sponsored by the World Bank, the FAO, The United National Development Programme, and the United Nations Environment Programme. Ten of the CGIAR Centers and Associated Centers have mandates to work on staple food crops. Research at these IARCs covers food commodities that provide 75% of the calories and protein requirements for developing-economy countries. The scientists responsible for carrying out this regional, crop-based research include virologists, plant pathologists, entomologists and weed scientists. Facilities include laboratories and greenhouses capable of pest diagnosis and research.
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Zilinskas, R. A., 'Confronting Biological Threats to International Security: A Biological Early Warning Program', ed Zilinskas, R. A., The Microbiologist and Biological Defense Research: Ethics, Politics, and International Security, (New York Academy of Sciences: New York, 1992), 146-176
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Mark Wheelis, PhD (Chair). Section of Microbiology, Division of Biological Sciences, University of California, Davis, California, USA
Pamela Anderson, DSc. Virology Unit, Centro Internacional de Agricultura Tropical, Cali, Colombia
Peter Barss, MPH, MD, DSc. Direction de la Sante Publique, Montreal, Quebec, Canada
Martin Hugh-Jones, VetMB, MPH, PhD. Department of Epidemiology and Community Medicine, School of Veterinary Medicine, Lousiana State University, Baton Rouge, Lousiana, USA
Matthew Meselson, PhD. Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, USA
Gerald Myers, PhD. Los Alamos National Laboratory, Las Alamos, New Mexico, USA
Meryl Nass, MD. Wing Memorial Hospital, Palmer, Massachusetts, USA
Barbara Hatch Rosenberg, PhD. Division of Natural Sciences, State University of New York, Purchase, New York, USA
Jack Woodall, PhD. Griffin Laboratory, NY State Department of Health, Albany, New York, USA
Raymond Zilinskas, PhD. Center for Public Issues in Biotechnology, University of Maryland Biotechnology Institute, College Park, Maryland, USA