OVERVIEW

Little guidance is currently available on biosafety considerations for conducting experimental field studies with microorganisms. While laboratory biosafety guidelines have been developed over a number of years in countries like Canada and the United States, these guidelines apply to the use of microorganisms in contained facilities. Less attention has been focused on biosafety guidance for environmental introductions of microorganisms in experimental field studies,

The following project was designed to develop generic biosafety guidance that could assist researchers when preparing for experimental field studies with microorganisms. The project reviewed available Canadian and international biosafety guidance pertaining to experimental field studies with microorganisms, and sought to develop a series of safe practices that could assist researchers in conducting field studies. A summary of the biosafety guidance, and examples of some safe practices, are provided in this poster.

INTRODUCTION

Safe practices are important when handling microorganisms to ensure protection of human health and the environment and to prevent damage to equipment and facilities and lost time due to accidents. Assessment of experimental field studies with microorganisms by regulatory agencies can ensure that the overall impact of a study will not cause harm to humans or the environment. However, regulatory assessment does not automatically ensure human and environmental safety from unexpected events that can occur during a field study. For example, if workers do not properly clean or sterilize utensils, equipment or clothing before leaving the site, a microorganism may be transported for long distances creating a potential for harm at new locations.

Responsibility for safety begins with the policies and attitudes of an institution which are reflected in the project director and workers. The project director must display appropriate concern for safety issues. Available biosafety guidance is essential for this purpose. The purpose of biosafety guidance is to identify safe practices related to conducting experimental field studies with microorganisms, which when properly considered, may prevent harm to workers, the general public, and the environment.

This project developed generic biosafety guidance that could apply to the conduct of a wide variety of experimental field studies with a microorganism including naturally occurring or genetically modified microorganisms, as well as complex mixtures of unidentified microorganisms in a consortium. Experimental field studies are studies conducted outside contained facilities for the purposes of research and development. These studies could be for a variety of purposes, such as to obtain data for efficacy tests or for environmental and human safety assessment purposes. While the guidance could apply to both small scale (less than 10 hectares terrestrial, less than one hectare aquatic) and large scale experimental field studies with microorganisms, experience should first be gained with a microorganism in small scale studies to provide a basis for safely conducting large scale studies.

It was determined that all experimental field studies will] have common features such as the need for proper planning and preparation, qualified and well trained workers, and a familiarity with the characteristics of the microorganism and the introduction site. The biosafety guidance developed in this project addresses safety issues in sequential stages from initial planning and laboratory activities supporting field studies, to microorganism introduction procedures and final field study termination procedures (Figure 1). The guidance draws upon the Laboratory Biosafety Guidelines (1996) developed by Health Canada and the Medical Research Council of Canada and extends some of the concepts and practices to field situations. It also draws upon other guidance prepared by various agencies in the United States and the United Kingdom.

SAFE FIELD STUDY PRACTICES

The safe conduct of any experimental field study with a microorganism requires attention to many attributes of the project. A thorough understanding is required of the characteristics of the microorganism, the introduction environment, and the method of application. As well, a thorough understanding is required of safety considerations at each stage in the conduct of an experimental field study. The safe practices in this biosafety guidance are organized around the following field study stages:

planning and preparation

transport of the microorganism to the site

Application of the microorganism

monitoring

security

contingency planning

termination of the study

Since the safe practices in this biosafety guidance could apply to a wide variety of potential microorganisms, applications and introduction environments, they are outlined in generic terms. However, certain safe practices may not always be strictly applicable. Experience and good judgment should be used in considering the safe practices for specific experimental field studies.

SAFE PRACTICE EXAMPLES

A. PREPARATION FOR FIELD STUDY

Related Laboratory Practices. The appropriate safe laboratory practices outlined in the Laboratory Biosafety Guidelines (1996) published by Health Canada should be followed.

Choice of Microorganism. Careful consideration should be given to avoid choosing a microorganism known to exhibit hazard characteristics such as pathogenicity unless this is absolutely necessary to fulfill the objectives of the field study. Any potential hazards and specific decontamination requirements associated with the microorganism should be well understood and taken into consideration when planning the field study.

Site Selection. The site should chosen to minimize exposure of the microorganism to humans, plants and animals around the site. Sites should be easily demarcated from surroundings.

Equipment Selection/Handling. Bioreactors, tanks, and other equipment containing the microorganism should be impermeable, resistant to corrosion, easy to clew and sterilize, and fittings should be well sealed.

Statistical Design. A statistician should be consulted in development of the experimental design to avoid poor designs necessitating repetition of experiments, and potential additional human and environmental exposure.

Site Preparation. An appropriate buffer zone should be established around the site. The site should be fully prepared prior to transport of the microorganism to site, and any necessary bioreactors or other equipment (liners etc.) should be in place and tested to meet manufacturer's specifications.

Personnel. Before the beginning of the study, a project director with appropriate qualifications, training, and experience should be designated to oversee the safety of the field study.

Record Keeping. All records should be kept in bound notebooks, with permanent black pen.

Public Notice. Persons situated close to the site should be advised of the study to be conducted and any restrictions on the site. Local authorities should be contacted to determine that appropriate regulatory requirements will be followed.

B. TRANSPORT OF MICROORGANISM

Microorganisms must be packaged, labeled and transported in accordance with the Transportation of Dangerous Goods Act and Regulations. Microorganisms should be transported in the safest means possible and by the shortest and most direct route to the field study site.

C. APPLICATION OF MICROORGANISM

Microorganism Introduction Procedures. An introduction protocol for the microorganism should be carefully followed and any deviation recorded. The microorganism should be applied in the minimum amount, frequency and duration necessary to meet the objectives of the study.

Equipment Operation and Maintenance. All equipment that comes into contact with the test microorganism should be stored separately from equipment used for other purposes. Depending on the hazard characteristics of the microorganism, appropriate disinfection practices should be followed before removing equipment from the field.

Operation of Bioreactors and Tanks. Operation of bioreactors and tanks should use the minimum aeration necessary and should avoid producing aerosols. Protective materials (liners, leachate control system) should be placed under bioreactors or tanks in order to contain releases and prevent leaching into groundwater.

Applications in Greenhouses. Access to the greenhouse should be limited and controllable. Windows should be kept closed and/or screened.

Protection Of Workers. Workers should wear protective clothing such as boots, gloves, and lab coats or suits, as appropriate to the potential hazard characteristic of the microorganism. Workers should not eat, drink, smoke, store food or utensils, apply cosmetics and insert or remove contact lenses around the site, especially at the time of microorganism application, tilling or sampling.

Storage of Microorganism. The storage area for the microorganism should be locked with limited access to the area.

Decontamination and Microorganism Disposal Procedures. All equipment, supplies etc., contacting the test microorganism should be sterilized no later than the end of the day. Efficacy of the sterilization procedure should be verified.

D. MONITORING

On-Site Monitoring. Regular observations should be made and recorded to determine adverse effects on workers and the environment at the site. Waste streams from the site should be monitored for the microorganism and any toxicant being treated and/or toxic by-products.

Off-Site Monitoring. Monitoring should be conducted periodically to determine if the microorganism is dispersing from the site. Any persons, plants or animals surrounding the site which could be adversely affected by the microorganism should be carefully monitored throughout the duration of the field study.

E. SECURITY

Access to the study site should be limited to authorized personnel only. Appropriate fences or other barriers to site access for unwanted visitors and animals should be considered for the field site.

F. CONTINGENCY PLANS

A contingency plan to handle responses to accidents and emergencies should be available to personnel at the field study site. Contingency plans should specify responses to potentially adverse situations including inadvertent spills or releases of the microorganism, weather conditions or flooding. The contingency plan should designate a Coordinator responsible for directing any emergency response measures and should specify arrangements for contacting local police, fire, hospital and regulatory agency authorities. All accidents should be reported immediately.

G. TERMINATION OF THE FIELD STUDY

Concentrations of the microorganism at the site should be determined at the end of the field study and, if necessary, attempts should be made to eliminate the microorganism from the site. An accurate and detailed final report summarizing the results and activities of the field study should be maintained on file.

CONCLUSION

The purpose of this project was to develop biosafety guidance for ensuring the safe conduct of experimental field studies with microorganisms. It was determined that all experimental field studies will have certain features in common, and that generic biosafety guidance could be developed to assist researchers in conducting safe experimental field studies. This poster provides a summary of the biosafety guidance and examples of safe practices. For more information, please contact Tom Edge, Institute of Biology, Carleton University, Ottawa, Ont., Canada K1S 5B6; (613) 520-2600 x3650.

Figure 1. Experimental field study stages.

i. Related laboratory practices

ii. Choice of microorganism

iii. Site selection

iv. Equipment selection/handling

A. Preparation for field study v. Statistical design

vi. Site preparation

vii. Personnel

viii. Record keeping

ix. Public notice

B. Transport of microorganism to site

i. Microorganism introduction procedures

ii. Equipment operation/maintenance

iii. Operation of bioreactors and tanks

C. Application of microorganism iv. Applications in greenhouses

v. Protection of workers

vii. Decontamination and disposal procedures

i. On-site monitoring

D. Monitoring ii. Off-site monitoring

E. Security

F. Contingency plans

G. Termination of study