(1) GILSP (Good Industrial Large Scale Practice)

This refers to the application of rDNA microorganisms at the minimum level of physical containment that satisfies the following criteria.

(i) The host cells must

1 Be non pathogenic to humans;

2 Be uncontaminated by exogenous factors (viruses, etc.) that are pathogenic to humans; and

3 Have either an extended history of safe use or be subject to intrinsic environmental limitations that permit growth in an industrial setting but that permit only limited survival without adverse consequences in outside applications.

(ii) The donor DNA and vectors must be

1 Well characterized and free from known harmful gene sequences;

2 Size-limited as much as possible to only the DNA required to perform the intended function; and

3 Poorly mobilizable and unable to transfer any resistant markers to microorganisms not known to acquire them naturally.

iii) rDNA microorganisms must

1 Be non pathogenic to humans; and

2 Not have higher propagative abilities than their host cells.

(2) Category 1

This refers to the application of rDNA microorganisms at a certain level of physical containment, excluding rDNA microorganisms that fulfill GILSP conditions and that are not pathogenic for humans.

(3) Category 2

This refers to the application of rDNA microorganisms at a certain level of physical containment, including microorganisms that are capable of infecting humans but that have a minimal likelihood of being pathogenic for humans even when handled directly, and for which sufficient preventive measures and effective therapy exist for use in case of infection.

(4) Category 3

This refers to the application of rDNA microorganisms that do not meet Category 2 conditions, that are pathogenic for humans, and that require very careful handling. Such applications are to be conducted at a specified level of physical containment in case of infection, and may be conducted only if hazards are relatively minimal, and if sufficient preventive measures and effective therapies exist.

In addition, rDNA microorganisms whose pathogenicity exceeds that specified in Category 3 applications shall be classified in a special category.

(1) Application in a simulated model environment

This refers to the experimental application of rDNA microorganisms classified under GILSP or Category-1 applications in Section 3-3. 1). Applications are to be conducted in a specifically restricted area under conditions that minimize the spread of rDNA microorganisms to the outside area and that minimize gene transfers from the rDNA microorganisms to other organisms in the outside area

(2)Application in an open system

This refers to the application in an open system of rDNA microorganisms whose safety has been confirmed in the simulated model environment.

4. Facilities and equipment usedfor rDNA microorganisms

Facilities and equipment used for applying rDNA microorganisms in GILSP, Category 1, Category 2, and Category 3 usages shall satisfy the conditions listed in the Appendix.

Facilities and equipment used for applying rDNA microorganisms in a simulated model environment should satisfy the following criteria:
1) There should be a work area clearly distinguished from other areas, and biohazard signs should be posted as necessary.

2) An appropriate confined field and management facilities should be established to utilize the rDNA microorganisms in the work area, taking into account their propagative properties, actual methods to limit their propagation, their physiological characteristics, and their application in an open system of the recombinant biota as well as the surrounding biota.

5. Handling of rDNA microorganisms

In applying rDNA microorganisms to GILSP, Category 1, Category 2 and Category 3 usages, or to a simulated model environment, rDNA microorganisms should be managed in compliance with the following:

1) Control of facilities and equipment for fermentation

(1) In GILSP or Category1 applications, the leakage of rDNA microorganisms should be minimized in seeding the combination to the cultivation or fermentation equipment, in sampling, or in transferring recombinants between cultures. In Category 2 or Category 3 applications, appropriate measures should be taken to prevent leakage of rDNA microorganisms; if any such leakage occurs, disinfection should be undertaken in a speedy and approved manner. In the case of application in a simulated model environment, measures should be taken as necessary to minimize the disposal of rDNA microorganisms from the work area.

(2) The leakage of aerosols from cultivation or fermentation equipment should be minimized in GILSP or Category 1 applications, and should be prevented in Category 2 and Category 3 applications.

(3) After completion of operations for employing rDNA microorganisms under GILSP or Category 1 applications or in a simulated model environment, the facilities and equipment used should be washed and disinfected; facilities used for Category 2 or Category 3 applications should be sterilized by fully approved methods.

Wastes should be disposed after inactivating them as required to attain the prescribed safety level. They should be inactivated by approved methods in Category 1 applications (including certain applications of rDNA microorganisms in a simulated model environment), and should sterilized by fully approved methods in Category 2 or Category 3 applications.

3) Storage

(1) Containers used to store materials, including rDNA microorganisms, should be clearly labeled as containing rDNA microorganisms and should be safely stored in specific storage facilities set up in advance, and especially so in the case of Category2 and Category3 applications. Signs describing applications as required levels, i.e., rDNA Microorganisms in storage (GILSP applications), rDNA Microorganisms in storage (Category 1 applications), rDNA Microorganisms in storage (Category 2 applications), rDNA Microorganisms in storage (Category3 applications), or rDNA Microorganisms in storage (Application in a simulated model environment) should be posted in conspicuous places in each storage facility.

(2) A catalogue of the stored materials harboring rDNA microorganisms should be prepared and maintained.

(1) If materials containing rDNA microorganisms are transported outside the work area, such materials should be placed in a container of sufficient strength, and the container should be sealed to prevent leakage of the contents.

In Category2 or Category3 applications, especially precautions should be taken so that the leakage of the contents does not occur hermetically, even if the container is damaged.

(2) The container to be used to transport the material harboring rDNA microorganisms should be clearly labeled on a conspicuous part of its surface with Handle with Care in red lettering.

(1) The facilities and equipment used for handling rDNA microorganisms should be given performance inspections immediately after their installation and periodically thereafter.

Sterilization equipment used in Category2 or Category3 applications should be inspected every six months.

(2) Whenever part of equipment used for such purpose as packing and sealing is/are modified or replaced, the airtightness and other functioning of the equipment should be examined.

(3) The airtightness of fermentation or cultivation equipment and related equipment used in Category 2 or Category 3 applications should be confirmed by fully approved methods during their operation.

(4) All facilities and equipment used in Category 2 and Category 3 applications should be identified by serial number, and their use should be strictly controlled.

(5) Sterilization equipment should be disinfected by approved methods on the occasion of replacement, periodic inspection, and modification of the apparatus itself.

(1) Signs indicating GILSP Applications, Category 1 Applications, Category 2 Applications, Category 3 Applications, or Applications in a Simulated Model Environment should be posted at each work area during work periods .

(2) Work areas should be kept clean; and, especially when utilizing rDNA microorganisms in industrial manufacturing processes, insects and rodents should be exterminated.

(3) Personnel should wear designated work clothing in the work area. Particular clothing should be used exclusively in the case of Category 2 or Category 3 applications. In addition, in Category 3 applications, personnel should completely change clothing and should take a shower when entering and leaving the work area.

Section 4. rDNA small laboratory animals

1. General matters

rDNA small laboratory animals whose safety has been confirmed should be utilized as described in Sections 4-3 and 4-4. However, in certain cases, the use of rDNA small laboratory animals should be exceptionally restricted.

The persons responsible for the use of rDNA small laboratory animals must obey the laws and ordinances relating to the utilization of laboratory animals; e.g., the Law concerning the Protection and Management of Animals (Law No. 105, 1973), the Ordinance concerning the Breeding and Custody of Laboratory Animals (Notice No. 6, Prime Minister's Office, 1980), and so forth.

2. Evaluation factors

1) Type of rDNA small laboratory animals employed

2) Host animals or the same biological species to which the hosts belong

(1) Taxonomic position

(2) Background information regarding utilization by humans and distribution in the natural environments

(3) Reproductive and propagative properties, as well as genetic characteristics

(4) Survival and reproductive abilities in natural environments

(5) Other physiological characteristics

(1) Identified/unidentified

(2) Structure and origin

(3) Functions of genes

(1) Names and origin

(2) Characteristics

(1) Methods of preparing rDNA smalllaboratory animals

(i) Structure and construction methods of rDNA molecules

(ii) Methods of introducing rDNA molecules into host cells

(iii) Developmental processes of rDNA small laboratory animals

(2) Location of rDNA molecules in host cells, and stability of the expression

(3) Differences between rDNA small laboratory animals and host animals or the same biological species to which the recombinant animals are derived.

(i) Reproductive and propagative properties, and genetic characteristics

(ii) Survival and reproductive abilities in natural environments

(iii) Production of infectious viruses

(4) Other physiological characteristics

3. Facilities and equipment used for rDNA small laboratory animals

Facilities and equipment used for the employment rDNA small laboratory animals should be installed to satisfy the following criteria.

1) Facilities and apparatus for rDNA small laboratory animals should be installed in a work area clearly distinguished from other areas. A work area may be formatted as the whole area for facilities in which rDNA small laboratory animals are bred (hereinafter referred to as breeding facilities).

2) Appropriate equipment should be set up to prevent the escape of rDNA small laboratory animals through doorways, ventilation ducts, drainage locations, and windows in the breeding facilities and the work area, taking into consideration the animals, reproductive and propagative properties and their physiological characteristics.

3) Cages and other equipment used for the breeding and maintenance of rDNA small laboratory animals (hereinafter referred to as breeding container) should not be easily openable by force, vibration, or the movement of the animals.

4. Handling of rDNA small laboratory animals

1) The breeding of rDNA small laboratory animals should be conducted in accordance with the following.

(1) Breeding of rDNA small laboratory animals

rDNA small laboratory animals should be bred in breeding containers clearly distinguished from other containers used for breeding non rDNA animals. In additional, it is desirable that rDNA small laboratory animals be individually distinguished; however, if individual distinction is difficult, rDNA small laboratory animals may be managed according to breeding group.

(2) Disposal of wastes relating to rDNA small laboratory animals

Wastes relating to rDNA small laboratory animals (including the carcasses of rDNA small laboratory animals) should be disposed of after sterilization or incineration, as necessary.

(3) Transporting rDNA small laboratory animals

(i) In transporting rDNA small laboratory animals outside the work area, containers possessing sufficient strength and structure to prevent escape the animals should be used.

(ii) The words Handle with Care in red lettering should be clearly displayed on all containers that contain rDNA small laboratory animals.

(4) Maintenance and management of facilities and equipment

Facilities and equipment used for handling rDNA small laboratory animals should be performance-tested at the time of installation and periodically thereafter, and their original performance level should be maintained.

(5) Other requirements

(i) A sign stating Application of rDNA small laboratory animals should be posted at each work area being used for the handling of rDNA small laboratory animals.

(ii) The work areas should be kept clean.

(iii) Working clothes should be worn in the work areas.

If rDNA small laboratory animals are to be transferred to other facilities and to other personnel, the person responsible must notify the receiving personnel of the following:

(1) That the animals being transferred are rDNA organisms; and

(2) That experiments employing rDNA small laboratory animals are subject to regulations specified in the Guidelines for Recombinant DNA Experiment (Decision of the Prime Minister on August 27, 1979) and the Guidelines for Recombinant DNA Experiments in Universities and Other Research Institutions

Section 1. Establishment of management systems by person responsible

The person responsible (This term applies to a person who is entrusted with the operation of applications in a simulated model environment) should establish management systems in compliance with the following, so as to ensure the safe application of rDNA organisms.

1. Appoint an operations administrator and a safety operations manager for each work site or institute where work will be conducted, and appoint replacements for the above operations administrator and safety operations manager to execute their duties in case of their absence due to illness or other reasons.

2. Establish a safe operations committee and appoint the members of that committee. The committee's members shall be required to periodically review the safety of the applications of rDNA organisms.

3. Oversee the operations administrator in the execution of the duties prescribed in Section 2 below.

4. Oversee the safety operations manager in the execution of the duties prescribed in Section 3-2 below.

Section 2. Operations administrator

The operations administrator should fully understand the following guidelines and should perform the duties detailed below.

1. Observe these guidelines in planning and executing operations, and appropriately manage and supervise each entire operation in close cooperation with the safety operations manager.

2. Prior to initiating operations, educate and train operations personnel regarding safety.

3. Post in a clearly visible location within the work area and storage facilities necessary information related to the handling of rDNA organisms.

4. Require, as appropriate according to the level of operations, that persons entering a work area be accompanied by operations personnel, and ensure that such persons obey the directions of the operations personnel upon entering a work area.

5. Make, and maintain for five years from the date of completion of an operation, records of the following items:

1) Reference names of each rDNA organisms and container number;

2) Storage and history of rDNA organisms employed;

3) The biological properties of rDNA organisms employed and the dates of their inspection;

4) Name and address, and the purpose of application, of organizations from which rDNA organisms were transferred (in the case of rDNA small laboratory animals, the date, number of animals, and the places of transfer also are required);

5) Results of health examinationsof personnel;

6) Records of reviews made by the Safe Operations Committee (including documents used as a basis for confirming that operating standards conform to the Guidelines); and

7) Records of the periodic inspections of facilities and equipment.

Section 3. Safety operations manager

1. The safety operations manager who is to assist the operations administrator should be appointed from among personnel who are knowledgeable about and acquainted with rDNA technology, so as to ensure safety in the application of rDNA organisms.

2. The safety operations manager should fully understand these guidelines and should properly discharge the following duties:

1) Ensure that operations are carried out in accordance with these Guidelines.

2) Advise or report information to the administrator as necessary .

3) Conduct any operations necessary to secure safety.

Section 4. Operation personnel

Operations personnel should properly discharge the following duties.

1. Understand the operations fully and maintain safety conducting them.

2. Give appropriate instructions to ensure full safety when any persons enter the work area without operations personnel.

Section 5. Safe operations committee

1. There should be formed a Safe Operations Committee consisting of members from appropriate fields, considering that members will need to process highly specialized knowledge of recombinant technology and organisms, as well as have overall general knowledge.

2. The committee should investigate and consider the following matters and advise the responsible person at his (her) request.

1) The suitability of handling methods for rDNA organisms;

2) The suitability of education, training, and health care regarding the safety of personnel in conducting operations;

3) Treatments necessary in case of accidents, and improvements of operating procedures that are advisable to present accidents;

4) Other factors necessary to secure of operating safety; and

5) The suitability of production and sale methods concerning rDNA small laboratory animals, in the context of the Law concerning the Production and Management of Animals and other laws and ordinances concerning the applications of rDNA small laboratory animals).

3. The committee may request reports from the operations administrator or the safety operations manager as necessary.

Section 6. Education and training of personnel

The operations administrator should ensure that the operations personnel fully understand these Guidelines and should ensure that such personnel receive appropriate education and training with regard to the following matters prior to initiating applications involving rDNA organisms.

1. Knowledge of safety levels of rDNA organisms being handled;

2. Knowledge of the proper handling of rDNA organisms according to their safety evaluation;

3. Knowledge of the proper use of facilities and equipment required for handling rDNA organisms;

4. Knowledge of the safety of operations to be conducted; and

5. Knowledge of measures to be taken in case of accidents.

Section 7. Health care

1. The person responsible should ensure that health examinations of operations personnel are conducted prior to their initiation of operations and at intervals of no longer than one year thereafter.

2. The person responsible should examine the measures to prevent and treat any infection or illness that could possibly arise from the engaging of personnel in Category 2 or Category 3 applications.

3. The person responsible should require operations personnel to undergo immediate health examinations and take other appropriate measures if it is the possible that personnel have been infected in conducting of Category 2 or Category 3 applications.

Blood-sera samples should be collected for operations personnel who will be engaged in Category 3 applications prior to the initiation of operations and should be stored for two years after such personnel cease work on the operations.

1. In order to ensure the safe application of rDNA organisms, the person responsible may request the Minister of Agriculture, Forestry and Fisheries to approve the safety criteria regarding facilities, equipment, and procedures utilized, to ensure compliance with these Guidelines.

2. The person responsible should collect information relating to rDNA organisms and their applications, and should report immediately to the Ministry of Agriculture, Forestry and Fisheries any new information that might influence the safety evaluation of rDNA organisms.

1. The person responsible should make efforts to accumulate sufficient knowledge to ensure the safety of rDNA microorganisms intended to be applied in an open system.

2. rDNA organisms whose hosts are animal cells (excluding rDNA organisms that have not progressed sufficiently to be differentiated as individuals) shall be kept in a specifically controlled environment for a specified length of time. If the person responsible requests the Ministry of Agriculture, Forestry and Fisheries to approve a safety evaluation of rDNA organisms whose hosts are animal cells, the approval shall be made on a case by case basis.

The species, characteristics, and purposes of applying rDNA animals shall be taken into consideration, and the regulations concerning their usage shall be applied appropriately.

3. At present, provisions relating to rDNA microorganisms in these Guidelines shall be applied to non cellular organisms (containing rDNA molecules) that are directly injected into plants or animals. However, plants or animals into which non cellular organisms are injected shall not be classified as rDNA organisms.

4. At present, a person responsible who intends to conduct sales of rDNA plants that might be able to propagate naturally in Japan, or who intends to produce foods, drinks, oils, or fats using such rDNA plants as materials, should confirm the initial safety of such plants as described in Chapter 2, Section 2-1 1).

5. Regarding matters not specified in these Guidelines, in certain applications the director general of the relevant bureau may establish necessary additionally provisions in order to implement of these guidelines.
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For further information, please contact: Innovative Technology Division Research Council Secretariat Ministry of Agriculture, Forestry and Fisheries Kasumigaseki 1-2-1 Chiyoda, Tokyo 100 JAPAN Telephone: Int.+81-3-3502-3919 Facsimile: Int.+81-3-3502-4028