Veterinary Biologics Guideline 3.2
Regulation of Biotechnology-Derived Veterinary Biologics

I. Introduction

The purpose of this document is to provide guidance to researchers, manufacturers and importers of biotechnology-derived veterinary biologics concerning the regulation of these products in Canada.

The Canadian Centre for Veterinary Biologics (CCVB) of the Canadian Food Inspection Agency (CFIA) is responsible for regulating the manufacturing, testing, importation and use of veterinary biologics in Canada, including veterinary biologics produced using modern techniques of biotechnology, under the authority of the Health of Animals Act and Health of Animals Regulations.

Some specific provisions concerning biotechnology-derived veterinary biologics are provided in section 120.4 of the Health of Animals Regulations. Prior to the "release" of a live genetically modified veterinary biologic, the researcher, manufacturer or importer is required to provide the CCVB with detailed information about the origin and nature of the novel biotechnology-derived veterinary biologic, in addition to the information required of conventional veterinary biologics. The release of a veterinary biologic is usually associated with the licensing of a product, but it might also be related to conducting a research trial under confined field conditions, or the emergency use of an unlicensed product.

To conform to Canada's environmental protection legislation, the Canadian Environmental Protection Act (CEPA 1999) and New Substances Notification Regulations, as well as the Health of Animals Regulations, before any novel veterinary biologic may be released from laboratory containment into the environment, the CCVB must assess all potential animal health, human health, and environmental aspects of the proposed release. For products of biotechnology, this regulatory review process involves the preparation of an environmental assessment (EA), a document that contains information on the molecular and biological characteristics of the recombinant organism, target and non-target animal safety, human safety, environmental considerations and risk-mitigating measures. Section V of the present guideline provides more information about the EA process, and provides an outline for preparing an EA.

Biotechnology-derived veterinary biologics are licensed in Canada based on the same four principles as conventionally produced veterinary biologics: purity, potency, safety and efficacy. However, due to the inherent variability of veterinary biologic products, and the added uncertainty surrounding a live genetically modified organism's characteristics, the specific studies that are required to demonstrate each of these fundamental properties depends on the product, its intended use, and the associated risks.

It is important to determine that the genes that are added, mutated, or deleted do not confer changes that compromise the safety characteristics of the vaccine organisms. A genetically manipulated microorganism might not always exhibit the expected characteristics; however, well-designed laboratory studies and contained animal trials should reveal any unplanned outcomes. Precautions must be taken to ensure that the genetic manipulations do not impart increased virulence, pathogenicity, or survival advantages in these organisms, beyond those found in natural or wild-type forms. The genetic modifications must not impart undesirable new or increased adhesive or invasive factors, colonization properties, different survival within the host, oncogenic properties, or other deleterious effects.

More information on the documents and data required to license a veterinary biologic in Canada, to conduct a field trial in Canada, or to obtain authorization to import or release an unlicensed veterinary biologic for emergency use, is available in other guidelines on the CCVB website.

A. Legal Authority

The Health of Animals Act and Health of Animals Regulations administered by the CFIA give authority to the CCVB to regulate the manufacturing, importation, release and use of veterinary biologics in Canada.

The Health of Animals Act provides the following definition:

• "veterinary biologic" means
  1. a helminth, protozoa or micro-organism,
  2. a substance or mixture of substances derived from animals, helminths, protozoa or micro-organisms, or
  3. a substance of synthetic origin
  • that is manufactured, sold or represented for use in restoring, correcting or modifying organic functions in animals or for use in the diagnosis, treatment, mitigation or prevention of a disease, disorder, abnormal physical state, or the symptoms thereof, in animals.

This definition provides the mandate to regulate any product that is represented as a veterinary biologic, whether produced by modern techniques of biotechnology, or by conventional methods.

Section 64(1)(s) of the Health of Animals Act provides the Governor in Council the authority to make regulations for prohibiting or regulating the importation, preparation, manufacturing, preserving, packing, labelling, storing, testing, transportation, sale, conditions of sale, advertising for sale, use and disposal of veterinary biologics and regulating their purity, potency, efficacy and safety.

In the Regulations, the provisions specific to live genetically modified veterinary biologics are the following:

• 120.3 (1) Subject to subsection (2), no person shall release a veterinary biologic unless the person
  1. submits an application for a permit for the proposed release to the Minister, accompanied by sufficient information, including the information referred to in section 120.4, to enable the Minister to determine whether the proposed release is
     1. unlikely to result in the introduction into Canada or the spread within Canada of any vector, disease or toxic substance, and
     2. unlikely to pose a risk of harm to the environment or to human or animal health; and
  2. is issued a permit for the proposed release under section 160.
• 120.4 (1) The accompanying information that is required to be provided by a person to the Minister pursuant to paragraph 120.3(1)(a) includes the following:
  1. in the case of a live genetically modified veterinary biologic
     1. a description of the donor organism and the methods of incorporation of the genes from the donor organism into the host, and
     2. a description of the live genetically modified veterinary biologic, including details relating to expression of the new gene and the stability of the incorporation of the new gene, and a comparison of the characteristics of the live genetically modified organism with those of the unmodified organism.

B. Definitions

Confined Release: a release of a veterinary biologic outside of containment where physical, chemical, operational or biological controls, or any combination thereof, are employed to restrict the exit or dispersal of the organism, or material from the organism, beyond a specified area.

Containment: a condition under which the movement of an organism is limited by one or more of the following mechanisms:

1. a set of standard practices that are generally used in microbiological laboratories;
2. special procedures, equipment and laboratory installations that provide physical and other barriers, which are applied in various degrees according to the estimated biohazard; and/or
3. biological barriers that limit either the infectivity of a vector or vehicle (plasmid or virus) for specific hosts, or its dissemination and survival in the environment.

Containment ensures that there is no release of an organism, or material from the organism, from a research facility to the environment.

Environment: the components of the Earth and includes the following:

1. air, land and water;
2. all layers of the atmosphere;
3. all organic and inorganic matter and living organisms; and
4. the interacting natural systems that include components referred to in paragraphs a. to c.

Live genetically modified veterinary biologic: a live veterinary biologic that contains or is made from an organism and is produced by recombinant DNA technology.

Release: any discharge or emission of a veterinary biologic into the environment.

II. Examples of Biotechnology-Derived Veterinary Biologics

Examples of biotechnology-derived veterinary biologics include the following:

• inactivated genetically engineered viral vaccines;
• bacterins produced from genetically engineered bacteria;
• viral or bacterial protein subunit vaccines purified from recombinant organisms or expression systems;
• plasmid DNA vaccines (non-replicating in eukaryotic cells);
• vaccines using a live viral vector to carry foreign genes (coding for immunizing antigens and/or immune stimulants);
• vaccines containing live organisms modified by gene mutation or deletion (no introduction of foreign DNA);
• vaccines containing live organisms modified by gene insertion or mutation (with the introduction of foreign DNA); and
• monoclonal antibody (hybridoma) products used prophylactically, therapeutically or as components of diagnostic kits. Note: The CCVB does not prepare a formal EA for monoclonal antibodies for in vitro use (e.g. in diagnostic kits), due to the limited risk associated with this type of biotechnology-derived veterinary biologic.

Note: Novel veterinary biologics produced by co-culturing organisms, serial passage using selective culture conditions to promote mutagenesis, or other less direct methods of genetic manipulation, may be regulated in a similar manner as biotechnology-derived veterinary biologics produced by direct, intentional genetic manipulation.

III. Licensing of Biotechnology-Derived Veterinary Biologics

As noted above, all veterinary biologics, including those produced by biotechnology, must be shown to be pure, potent, safe, and efficacious in order to become licensed for use in Canada. The assurance of safety for any veterinary biologic requires that the product does not harm animal or human health, and does not have adverse effects on the environment. The rigorous review of the methods used in the production and testing of the veterinary biologic, including the construction of any recombinant organisms, and the preparation of the EA can be considered to constitute part of the supplemental safety assessment for biotechnology-derived products.

The supplemental information required by the CCVB as a prerequisite to licensing a biotechnology-derived veterinary biologic product may include (but is not necessarily limited to) the following:

• information about the parental and donor organisms, including their origin/isolation history, pathogenicity, host range, tissue tropism, and natural environmental distribution;
• genetic modification procedures used to construct the altered organism, including information on any intermediate cloning vectors and screening techniques for identification;
• data on the molecular and biological properties of the master seed;
• sequencing data for the recombinant organism, including sequence alignment (percent identity) with parental and donor sequences as appropriate;
• description of any selectable markers, including antibiotic resistance genes;
• phenotypic characterization of the genetically engineered organism;
• in vitro and in vivo data to demonstrate the genetic and phenotypic stability of the construct;
• data demonstrating the purity of the master seed;
• information on the engineered organism's virulence traits, replication competency, infectivity, tissue tropism, and other factors affecting pathogenicity;
• information on the fate of the product when injected into the vaccinee, including its ability to multiply and be shed from the organism;
• an assessment of the recombinant organism's ability to affect non-target species;
• data on the ability of the organism to spread from vaccinated animals to other organisms and maintain itself in target and non-target animal populations;
• potential for horizontal gene transfer or recombination;
• survivability of the shed organism or spilled vaccine in the environment;
• data to support the safety of the genetically engineered products in target and non-target animal species;
• considerations and mitigative actions with respect to human safety;
• non-reversion to virulence studies examining the effect of multiple back-passages in the target animal species;
• data from immunogenicity and/or vaccination-challenge studies demonstrating efficacy; and
• results of studies validating the proposed test procedures for potency testing.

The specific data requirements will depend on the nature of the product, its intended use, and the associated risks. Due to the potential risks associated with the release of live biotechnology-derived veterinary biologics compared to inactivated biotechnology-derived organisms, plasmid DNA or purified recombinant protein-based vaccines, the CCVB requires more extensive documentation regarding the origin and nature of these live products when evaluating requests for authorization to release this type of veterinary biologic into the environment.

Information about the process for licensing veterinary biologics in Canada is available on the CCVB website. In particular, manufacturers are advised to review Veterinary Biologics Guideline 3.1: Guidance for Preparation of New Product Licensing (Registration) Submissions for Veterinary Biologics.

IV. Environmental Release of an Unlicensed Veterinary Biologic

The CCVB additionally evaluates requests to release unlicensed veterinary biologics in Canada for restricted use in field studies and emergency situations, as noted below.

A. Field Trials

In order to generate the required information for product licensing, in the latter stages of the product evaluation process, most veterinary biologics developed in Canada will need to be tested outside of the contained laboratory in a confined field trial. CCVB approval is required for product testing studies that involve the release of an unlicensed experimental veterinary biologic from laboratory physical containment. The manufacturer should provide the CCVB with a product licensing submission before proceeding to a stage where approval for a limited field trial outside the controlled containment facilities is sought. The licensing submission should include data from laboratory research and development studies, and from controlled experiments performed in containment, which fully characterize the recombinant organism and demonstrate its safety in target and key non-target animal species.

Veterinary Biologics Guideline 3.29: Safety Requirements for Veterinary Biologics contains information regarding field trials conducted to generate safety data for product licensing, including a list of the documents and data to be included in an application for a field trial.

Following receipt of a submission made by a researcher or manufacturer for permission to conduct a field trial in confinement, the CCVB evaluates the proposed release activity and undertakes a comprehensive consideration of the potential environmental effects and risks to human and animal health. The CCVB then prepares an EA summarizing the molecular and biological characteristics of the biotechnology-derived organism, and the target and non-target animal safety, human safety, and environmental considerations (refer to Section V). Any risk-mitigating measures are additionally described in the EA. Prior to authorizing the field trial, the CCVB may require additional assurance tests on the product to be performed by a laboratory acceptable to CCVB. The CCVB will also review and provide comments on the study protocol.

If the proposed field trial is approved, CCVB issues the manufacturer or researcher a Permit to Release Veterinary Biologics.

Occasionally, veterinary biologics produced in a foreign country also undergo field testing in Canada. In this case, the CCVB will evaluate the proposed confined release before allowing the importation of the test product.

An inspector of the CFIA may inspect the testing facility and may also monitor the study to verify compliance and ensure that permit conditions are being met.

Confined field trials are to be conducted under quarantine conditions acceptable to the CCVB, where there is adequate evidence of biological and/or physical control of the genetically engineered organism. Facilities shall be maintained and operated in an appropriate manner, to prevent the dissemination of the unlicensed veterinary biologic or animals that were exposed to the product. Test animals receiving any experimental veterinary biologic must not enter the food or feed chain without permission from the CCVB.

B. Emergency Use

Under certain circumstances (e.g. if there is an outbreak of a disease in Canada for which there is no effective licensed veterinary biologic available), the CCVB may consider an application for the release of an unlicensed biotechnology-derived veterinary biologic for emergency use. In these situations, the CCVB will require information on the origin and nature of the product, together with data demonstrating its purity, potency and safety, in order to fully evaluate the proposed release and prepare an EA (refer to Section V). In addition, the CCVB may request data supporting the efficacy of the veterinary biologic in order to be confident in its risk-benefit analysis. If the emergency use of the unlicensed veterinary biologic is approved, the CCVB will issue a Permit to Release Veterinary Biologics and/or a Permit to Import Veterinary Biologics, as applicable, with any necessary conditions and restrictions listed on the permit(s).

V. Environmental Assessment

A. General

The environmental assessment (EA) contains information on the molecular and biological characteristics of the biotechnology-derived organism, target animal and non-target animal safety, human safety, environmental considerations and risk mitigation measures. The manufacturer must submit all relevant data to aid the CCVB reviewer in preparing the EA. The CCVB reviewer will also independently research any potential safety issues, and may consult other federal and provincial government departments with expertise in areas of concern. For instance, potential human health and safety risks may be assessed in collaboration with Health Canada. An EA must be completed before the CCVB will authorize the release of a novel organism into the Canadian environment.

The scheduling of the Health of Animals Act and Health of Animals Regulations under Schedule 4 of the Canadian Environmental Protection Act, 1999 (CEPA 1999) exempts new biotechnology-derived veterinary biologics regulated by the CCVB from additional notification and assessment under the CEPA 1999. This scheduling is recognition that the environmental assessment authorities within the Health of Animals Act and Health of Animals Regulations provide an adequate level of regulatory control. Consequently, the CCVB is fully responsible for the environmental, human health and animal health assessment of a novel veterinary biologic product.

After the CCVB has completed the draft EA for a biotechnology-derived veterinary biologic, the manufacturer is provided a copy of the document and given the opportunity to identify any confidential business information. A publicly available version of the EA, which omits this confidential business information, is subsequently posted on the CFIA website, where it is accessible to the general public.

The public versions of previously completed EAs can be found on the CCVB's Environmental Assessments web page.

B. Outline for the EA

I. Introduction

• Proposed action - objectives, location and protocols
• Background - information on problem and action

II. Purpose and need for proposed action

• Significance - agricultural, scientific
• Rationale

III. Alternatives

• Available choices and their relative scientific merits
• Selection criteria and weight given to each alternative
• Justification for selection of proposed action

IV. Molecular and biological characteristics of parental and recombinant organisms

• Identification, sources, and strains of parental organisms
• Source, description, and function of foreign genetic material
• Method of accomplishing genetic modification
• Genetic and phenotypic stability of vaccine organism
• Potential for recombination and horizontal gene transfer
• Host range/specificity, tissue tropism and shed/spread capabilities
• Comparison of the modified organism to parental properties
• Route of administration/transmission

V. Human Safety

• Previous safe use
• Probability of human exposure
• Possible outcomes of human exposures
• Pathogenicity of parent microorganisms in humans
• Effect of gene manipulation on pathogenicity in humans
• Risk associated with widespread use of the vaccine

VI. Animal Safety

• Previous safe use
• Fate of the vaccine in target and non-target species
• Potential for shed and/or spread from vaccinate to contact target and non-target animals
• Reversion to virulence resulting from back passage in animals
• Effect of overdose in target and potential non-target species
• The extent of the host range and the degree of mobility of the vector
• Relative safety when compared to conventional vaccines
• Safety in pregnant animals and to offspring nursing vaccinated animals

VII. Affected Environment

• Extent of release into the environment - identify site
• Persistence of the vector in the environment/cumulative impacts
• Extent of exposure to non-target species
• Behaviour of parent microorganisms and vector in non-target species
• Physical and chemical factors which can affect survival, reproduction and dispersal of the vector

VIII. Environmental Consequences

• Risks and benefits - analysis of potential risks compared to benefits of proposed action/vaccine
• Relative safety compared to other vaccines

IX. Mitigative measures

• Worker safety
• Non-worker human safety
• Handling of vaccine
• Handling vaccinated or exposed animals
• Other

X. Monitoring

• General
• Human
• Animal

XI. Consultation and Contacts

XII. Conclusion and Actions

XIII. References

• List of all references cited or relied upon, including personal communications

XIV. Appendices

• Tables, figures and maps
• Reports