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RG-1 Regulatory guidance:
Chapter 2 - Data requirements for single ingredient approval and feed registration

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2.1 Process to streamline approvals for category 1 feed registrations

Purpose

The Canadian Food Inspection Agency (CFIA) and the feed industry continue to seek ways to streamline processes to help reduce the total time required to approve feed products requiring registration.

Background

Category 1 Feed Registrations are standard feeds that do not require safety or efficacy data review. These have relatively simple requirements and a straightforward approval process; however, they can take significant time to be evaluated due to the quality of submissions and subsequent interactions to ensure that information needed to evaluate the product is complete. This results in delays which affect all other registration submissions in the queue and contributes to a backlog.

Preliminary screening steps are currently built into the feed registration approval process so that when CFIA reviewers receive application packages, they are complete. These additional steps in the process can increase the overall time to register an application.

The streamlined process described in this document focuses on increasing the quality of registration submissions for Category 1 Feed Registrations to decrease the overall time it takes to review these products. Applications identified as streamlined will skip the preliminary technical screening, and proceed directly to the technical review to ensure the streamline requirements are met.

Benefits

Product eligibility

Products eligible for the streamlined process include:

The following product types are not eligible for the streamline process:

Requirements

The process is designed to meet the requirements of the Feeds Regulations, 1983. Eligible Category 1 feeds, must be labelled and contain the nutrient guarantees per Schedule I Table 3. Vitamin and mineral guarantees will be compared with minimums and maximums provided in Schedule I Table 4.

The requirements for the streamlined process have been organized into checklists found below:

Requirements and Checklist for Streamlined Approvals of Category 1 Feeds

Tools have been developed to assist the industry in demonstrating that requirements have been met and are available on the Animal Nutrition Association of Canada (ANAC) website at Streamline Approvals for Category 1 Feed Registrations

Note: applicants must self-identify that their application meets the requirements of the streamlined process. Identified applications that fail to provide all necessary information or that do not meet the requirements for the streamlined process, will be returned to the applicant and the applicant must begin a new registration application. Applications will not be placed on hold pending additional or revised information.

Process

The registration process would be as follows:

  1. registration package is completed and verified by the registrant against the requirements
  2. this includes verifying that every required element (as per CFIA's checklist) is complete, meets regulatory requirements, and is included in the package in the same order as proposed on the checklist
  3. the registrant must:
    • apply via the online service My CFIA
    • submit the application package as per chapter 1.3
    • select the permission type in My CFIA: Registration or approval
    • write in the online application name "Streamlined category 1 application…"
      • omitting to add this information to the application name may result to a longer evaluation process
    • upload the label, the completed and signed checklist, calculation sheets and all other required information in the appropriate sections
  4. the CFIA will review the file and either
    • issue a registration for applications that are complete and meet the requirements of the streamlined process
    • return the file if the application is incomplete
  5. the registration decision will then be sent to the applicant and the application fee will be retained by the CFIA

Note: this is a voluntary process. Products can still be submitted for registration without using the streamlined process, in which case traditional approval processes and timelines will apply. Before being submitted to CFIA for evaluation, ANAC Members can choose to have their registration packages pre-screened for completeness by ANAC – visit Streamline Approvals for Category 1 Feed Registrations.

Requirements and checklist for streamlined approvals of category 1 feeds

1. New and significant changes to standard mixed feeds listed in items 1-6 of Schedule I table 3 of the Feeds Regulations, 1983 - complete feeds (including milk replacers), supplements, macro-premixes, mineral feeds, micro-premixes, trace mineralized salts, or converter feeds

Requirements Page # or N/A CFIA use

Completed application package is submitted in the same order as per this checklist and as per RG-1 Chapter 1 – Administrative Requirements for Pre-market Assessment and Product Registration of Livestock Feed

  • Cover letter– indicating that this is a streamlined application
  • Application form
  • Application fee
  • Checklist

Label (3 copies) containing the information as per Section 26 of the Feeds Regulations, 1983 (see list in Appendix 1)

Note: for mixed feeds listed in items 1-6 of Schedule I Table 3, if label guarantees that are not indicated in Schedule I Table 3 appear on the label or if the label highlights the inclusion of another mixed feed, these feeds are not eligible for streamlined review

For example statements like – "Contains Moore's Mineral Complex" or "with Smith's Vitamin D".

Claims that are part of the Permissible claims policy for livestock feed are allowed on the label if the information to substantiate or support the permissible claim is included in the application package.

For complete feeds:

calculations showing that the nutrients requiring labelling guarantees as per Schedule I Table 3 meet or exceed the Schedule I Table 4 minimums and do not exceed the maximums

For mixed feeds other than complete feeds:

  • calculations showing that the nutrients requiring labelling guarantees as per Schedule I Table 3, when fed according to the directions for use, provide at least 10% of the Schedule I Table 4 nutrient minimum and do not exceed the nutrient maximums
  • if a nutrient does not provide at least 10% of the minimum value in Schedule I Table 4, the nutrient should not appear as a guarantee on the label
  • a calculation must still be included to demonstrate the rationale for excluding the guarantee

For medicated feeds:

calculations showing that the level of medication supplied is in accordance with the Compendium of Medicating Ingredient Brochure (CMIB) directions for use associated with the appropriate claim

Note: imported medicated feeds are not eligible for streamlined review

For new feed applications and all imported feeds:

  • a complete list of ingredients identified by their generic name and item number as listed in the Administrative Schedule IV or V of the Feeds Regulations, 1983, including Canadian Feed Registration Numbers for Part II ingredients or registered mixed feeds, must be included with the submission (on a separate sheet or on the label)
  • the complete list of ingredients should include all of the ingredients used in any premix or supplement used to make the category 1 feed (for example, an in-house premix)
  • if that premix or supplement is registered, the list of its ingredients is not required, only the registration number and a copy of the label
  • provide a copy of the label of any other registered feeds included in your mixed feed
  • the list of ingredients is not required if the streamlined application is for a renewal or an amendment to an already registered feed.

2. Simple mixtures and dilutions of single ingredient feeds only containing ingredients listed in schedule IV part I classes 1 to 7.1

Products admissible for streamlined approvals are:

Requirements Page # or N/A CFIA Use

Completed application package is submitted in the same order as per this checklist and as per RG-1 Chapter 1 – Administrative Requirements for Pre-market Assessment and Product Registration of Livestock Feed

  • Cover letter– indicating that this is a streamlined application
  • Application form
  • Application fee
  • Checklist

Label (3 copies) containing the information as per section 26 of the regulations (see list in Appendix 2)

Note: for simple mixtures and dilutions of single-ingredient feeds from Schedule IV part I Classes 1-7, only label guarantees indicated in the individual single-ingredient feed definitions should be present on the label

  • some single ingredient feeds allow for identifying the names of pelleting aids, mould inhibitors, anticaking or flow agents used in their manufacture
  • the names of these may be transferred to the label of these simple mixtures or dilutions of single-ingredient feeds
  • if they are registered, provide a copy of the label of any registered feeds included in your simple mixture
Calculations
  • calculations showing the amalgamation of guarantees being transposed from the single ingredient feeds

Appendix 1 – Label requirements for new and significant changes to standard mixed feeds listed in items 1-6 of schedule I table 3 of the Feeds Regulations, 1983

Proposed label

Appendix 2 – Label requirements for simple mixtures and dilutions of single-ingredient feeds only containing ingredients listed in schedule IV part I classes 1 to 7.1

Proposed label

2.2 Registration requirements for specialty products

For further guidance on some specialty feed products' registrations, refer to Chapter 3.

Administrative requirements

1. Standard registration requirements as indicated in Chapter 1.

2. Proposed labels must reflect the purpose of the product and the standards, packaging and labelling requirements of the Feeds Act and Regulations. Guarantees for nutrients and/or active ingredients must appear on the product label. Five copies of the proposed label and a copy of the product formulation (for example, percentage composition) are required for submission.

Product description and supporting information

3. A complete list of ingredients identified by generic name is required, as listed in Schedule IV or V of the Feeds Regulations. Registration numbers for Part II ingredients, mixed feeds, and other registered products must also be included.

Mixed feeds may contain ingredients that are not listed in either Schedule IV or V of the Feeds Regulations. If the applicant wishes to use a currently unapproved feed ingredient as part of the product formulation in question, it is also necessary to submit an application for registration to have the ingredient approved for use in livestock feeds in Canada prior to submitting the application package for the specialty product in question. Information concerning the application process for new feed ingredients can be found in section 2.3. "Single Ingredient Feed Evaluation Requirements". Note that the application for registration for the specialty product cannot be approved unless all of the ingredients used in the product formulation are approved and listed in either Schedule IV or V of the Feeds Regulations (that is, the application will be placed on hold).

4. The intended purpose of the feed (to assist in designating the name of the feed, for example, anion/cation balance feed, flavouring agent, antioxidant, forage additive) is also required.

5. Scientific studies must be presented in support of each label claim. These investigations must meet the following criteria:

6. Certificates of analysis are required from three different and recent lots of product, in addition to an accompanying analytical methodology to substantiate the guarantee(s) that appear on the product label.

7. Quality control procedures demonstrating product consistency are required, if applicable.

8. Product sample(s) (for products such as for forage additives, direct-fed microbials and enzyme supplements) may be requested by the evaluation officer or Feed Laboratory to verify analytical methodologies and/or label guarantees.

9. Stability (that is, shelf life) information must be submitted, if applicable. To support product stability, it will be necessary to submit certificates of analysis substantiating the guaranteed shelf life on the label (that is, at the date of manufacture and at the expiry date) for a minimum of three different lots of the product. The certificates of analysis for the level/concentration of the active ingredient(s) at the date of manufacture and at the expiry date must be for the same lot of product. The storage conditions used in the shelf life study should reflect the recommended storage conditions as stated on the product label.

Safety information

10. As outlined in sections 2.3 and 2.4 scientific investigations supporting product safety may include, but are not limited to: chemical analyses and/or harmful residues and/or toxicological evaluation and/or animal feeding studies and/or tissue residue analysis. The scientific studies presented in support of product safety must be:

11. Suitable methodologies for the detection of significant amounts of any ingredient, nutrient, compound, substance, or organism intentionally incorporated into the feed or occurring as a contaminant must also be provided.

Requirements for amendments (significant change) to specialty product registrations

Applications for significant change will be evaluated on a case by case basis. Changes including, but not limited to, the following product characteristics require the submission of efficacy and/or safety data (which meet the criteria previously outlined), as part of the application for registration amendment:

Please refer to Chapter 1 for additional information regarding significant changes to product registrations. The application fees for these types of changes are the same as those for a new application for registration for a Category 2 product.

Formulation changes involving ingredients that are not considered to be active ingredients or a significant source of nutrients (for example, flavours, carriers), will be evaluated on a case-by-case basis and may or may not require the submission of efficacy and/or safety data. However, it is recommended that the Feed Program be contacted ahead of time in order to determine what type of information and documentation will be required for submission with the proposed application for registration amendment.

Registration requirements for private labelled products

The private label registration process has been put in place to allow companies to commercialize a product that is already registered by another company under a different brand name. Such applications are considered standard feeds and do not require any additional safety or efficacy evaluations. Applications for registration of private labels must also include a letter from the company holding the original registration, signed by a person having signing authority for this company, allowing the private label applicant to use the information on file to register their product as a private label.

As the formulation of the private label product must correspond exactly to the formulation originally approved for the parent product, any mandatory information present on the original label (for example, list of ingredients, guaranteed analysis, directions for use, caution statements) must be present on the private label. If the private labelling company also manufactures the feed, the formulation is required and certificates of analysis may be requested.

Applications for amendments to private label registrations will be evaluated on a case-by-case basis according to the following criteria:

  1. Mandatory amendments

    For mandatory registration amendments (that is, amendments initiated by the Feed Program), the Feed Program will inform all holders of a registration for the type of product affected by the amendment, including private label registrants. Mandatory amendments must be made to both private labels and parent product labels. There will be no charge for such amendments.

    In cases where an application is made for a private label for which the status of the original product registration would not satisfy new mandatory requirements (due to concerns with efficacy and/or safety data), the private label application will be refused until the original product meets all registration requirements. The holder of the original registration will be notified of the situation.

    Minor mandatory amendments required by the Feed Program will be made to a private label product immediately upon registration or registration renewal, even though the label for the parent product may not yet comply with the new amendment. In these cases, the private label will be corrected before the original product label. Minor mandatory amendments to the original label will be made at the time of renewal or when an application for significant change is submitted to the Animal Feed Division.

  2. Voluntary amendments

    When the original registrant is granted an amendment (for example, revised guarantee, addition of a claim, changes in the formulation), the original registrant is responsible for notifying the private label registrant(s) of these amendments. In such cases, private label registrants are not obliged to update their registration to reflect the change(s) made to the original registration as long as the private label continues to comply with the Feeds Regulations. However, should the private labelling company chose to include the new information, an application package for a significant change must be submitted.

  3. Registration renewal

    A copy of the product formulation (including registration numbers for Part II ingredients, flavours, registered mixed feeds, etc.), and a letter from the company holding the registration of the parent product allowing the private label company to continue to use the information on file to renew the private label product, must be submitted with each application for registration renewal.

    Note: If a parent product is issued a temporary registration, the private label product will only be granted temporary registration status. The expiry date for the private label product will be the same as that for the parent product. The granting of permanent registration status to a private label product that was previously given a temporary registration is contingent upon the parent product receiving permanent registration status. If the application for permanent registration is submitted at the time of renewal, the registration fee will be in addition to the renewal fee. If the application is submitted prior to the expiration of the temporary registration, the registration fee is the same as that for the consideration of a new application for registration.

2.3 Single ingredient feed evaluation requirements

Single ingredient feed evaluation may require (but is not limited to):

2.4 Generic data requirements for safety evaluations of single ingredients

In order to facilitate the health & safety assessments of single ingredients, the following list of data requirements has been developed to clarify section 2.3.

These requirements will apply to all single ingredients which are not already approved for use in Canada. It is expected that all manufacturers/registrants will be able to supply the information listed if applicable. In the event that a preliminary evaluation identifies concerns, more specific information may be requested.

Single ingredients already cleared may be subject to a similar assessment if any doubt as to their safety or improper use is suspected.

Please note that information is required of active ingredients, major metabolites, and any contaminants of concern.

  1. Identification and Use of the Product
    • Name
    • Other names (i.e. international feed name, chemical name, botanical name, etc.)
    • Suggested rates, timing, intervals of feeding, withdrawal times
    • Identification of species of intended use
    • Unit amount
    • Product Label and Material Safety Data Sheet (MSDS) for product or ingredients
    • Outline of manufacture
  2. Chemical Components Including Impurities
    • Chemical name and synonyms
    • Chemical formula (molecular and structural)
    • Chemical Abstract Service (CAS) Number
    • Concentrations in the finished product (% or ppm)
    • Criteria of chemical identity and purity
    • Estimated shelf life
  3. Method(s) of Analysis, Recovery and Detection Limit Data for the Analyses (per section 6.2)
  4. Physicochemical Data (if applicable)
    • Molecular weight
    • Physical state
    • Density/specific gravity
    • Refractory index
    • Melting point
    • Boiling point
    • Flash point
    • Auto-ignition temperature
    • Vapour pressure
    • Vapour density
    • pH
    • Solubility in water
    • Octanol-water partition coefficient
    • Solubility in other solvents
    • Incompatibility
    • Polymerization
  5. Pertinent Livestock Toxicity Data
  6. Livestock Metabolic Fate and Residue Studies
    • Metabolic fate studies, to include analytical methods for recovery and detection limits
    • Residue studies for the parent compound and its possible metabolites, to include analytical methods for recovery and detection limits
    • Excretion data
    • Suggested Maximum Residue Limit(s) or tolerance
  7. Mammalian Toxicological Data

    Typical minimum mammalian toxicology data requirements are bolded. The additional studies or data may be required of particular products or constituents if deemed necessary.

    • Routes of entry
    • Rate and degree of absorption
    • Estimation of exposure
    • Acute toxicity: acute median lethality, skin/eye irritation, skin sensitization, mutagenicity (with and without activation)
    • Short-term toxicity (e.g. 28-day study)
    • Teratogenicity
    • Carcinogenicity
    • Developmental toxicity
    • Reproductive toxicity
    • Epidemiological studies
    • Chemical interactions
  8. Environmental Fate and Effects
    • Vapour pressure and volatilization
    • Hydrolysis
    • Photodegradation
    • Solubility in water
    • Octanol-water partition coefficient
    • Adsorption-desorption
    • Leaching
    • Biotransformation in soil (aerobic/anaerobic)
    • Biotransformation in aquatic systems (aerobic/anaerobic)
    • Toxicity to wildlife
    • Toxicity to aquatic organisms
    • Toxicity to soil organisms

Note: If tissue residue levels above background are evident, the Food Directorate of Health and Welfare Canada may require a more extensive review before approval for use is granted.

2.5 Data requirements for product safety evaluations: Explanatory notes

This section provides an explanation of the data requirements for safety evaluations of livestock feeds. It is to be used in conjunction with Section 2.3 "Single Ingredient Feed Evaluation Requirements" and Section 2.4 "Generic Data Requirements for Safety Evaluations of Single Ingredients" of the RG-1 Regulatory Guidance document.

We have attempted to make these notes useful to those who wish to obtain a general understanding of the data requirements, and to aid in the development of a complete data package. A listing of standard protocols used to develop the technical information is also provided for your convenience.

Questions should be forwarded to:

Animal Feed Division
Canadian Food Inspection Agency
59 Camelot Drive
Ottawa ON  K1A 0Y9
Fascimile: 613-773-7565 or 613-773-7566

1.0 Identification and use of the product

For each feed ingredient or product, please identify the following:

A) Name and Synonyms

The name of the product as it appears on the product label or bill of lading. Commonly used alternates or acronyms, including names used in other countries, most common chemical name, a botanical or species identification, etc.

B) Description of Use

Describe the intended use of the product in feed. The type of information required includes:

C) Species of Intended Use

Identify all livestock for which the product is intended.

D) Unit Amount

Identify what quantity is the product to be sold, transported and stored (e.g., 50 L metal drum, 20 kg plastic-lined canvas sack).

E) Product Label

Five (5) copies of the product label or bill of lading must be provided. Guidelines for labelling requirements of livestock feed can be found in Section 4.1 of the RG-1 Regulatory Guidance document.

F) Material Safety Data Sheet (MSDS) for Product and Ingredients

A MSDS is a comprehensive technical bulletin containing detailed information on a substance or product. It is a basic source of information for preliminary safety evaluations identifying items which may need to be examined in greater detail. The Workplace Hazardous Materials Information System (WHMIS) provides criteria for developing a MSDS. Please refer to Section 7.1 of the RG-1 Regulatory Guidance document "Responsibilities Regarding the Labelling of Feed Products under the Workplace Hazardous Material Information System (WHMIS)".

G) Outline of Manufacturing Process

Please provide a detailed description of the production and formulation processes, identifying all raw materials, processing times and temperatures, chemical and physical treatments including further processing steps, and other parameters which may influence the specifications, quality, or safety of a product.

A flow-chart diagram accompanying the description is recommended.

2.0 Chemical components including contaminants and impurities

A) Name and Synonyms

The chemical name and synonyms of all ingredients should be listed.

Resources

B) Contaminants and Impurities

Contaminants and impurities that are inherent to this product or introduced through processing should be identified and characterized.

C) Chemical Abstracts Service Registry Number

An identification number is assigned by the Chemical Abstracts Service (CAS) to differentiate between known chemicals. This is a useful reference when searching for technical information and should be provided for each ingredient.

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D) Chemical Formula (Molecular and Structural)

The molecular formula identifies the basic elements (atoms) of molecules. A structural formula is a diagram of the bonds between atoms. Comparing these with the formulae of substances whose properties are known is useful in anticipating hazards of substances which have not been tested. If isomeric mixtures exist, the ratio of isomers should be included, since different isomers may have different toxicities. For polymers (long chains of molecules), the structural formulae should show the repeating units along with the identification of links and cross-links.

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E) Concentration of Components

The amount of all ingredients, contaminants and impurities should be expressed as a percent by weight of the final product or by parts per million (ppm).

F) Criteria for Identity and Purity

The exact formulation of a particular product may vary, depending upon the manufacturer. A precise description of ingredients, including physical/chemical properties and contaminants or impurities, is needed to properly assess product safety. The methods for measuring the ingredients and/or contaminants in the product must be stated. Examples of acceptable analytical methods for component detection include proximate analysis, nuclear magnetic resonance (NMR), mass spectrometry, chromatography, etc.

Please refer to Section 6.2 of the RG-1 Regulatory Guidance document for the information required about the analytical methods being applied by the applicant/registrant.

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G) Estimated Shelf Life

This is the length of time the product can be stored without significant alterations to its chemical or biological composition. This includes specified storage times under ideal conditions, a description of the factors affecting shelf life, what happens when the product undergoes degradation or transformation, how one can tell if degradation has occurred, hazards associated with exceeding shelf life, and how the manufacturer has substantiated its estimation of shelf life.

Shelf life can be determined based upon industry standards, quality assessment, and compositional assessment.

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3.0 Method(s) of analysis, recovery, and detection limit data for the analyses

Acceptable analytical methods are required both to establish the ingredient purity (including contaminants), and for the ability to detect the ingredient in feed at its intended use rate. Methods must establish statistically relevant recovery and detection limits, and must allow for third-party laboratories to identify and quantify the active ingredients. For specific requirements of analytical methods, please refer to Section 6.2 of the RG-1 Regulatory Guidance document.

Other Resources

4.0 Physicochemical data

Physicochemical data is used to identify and differentiate between substances or products and to assist in predicting or determining the behaviour of substances in the human body, target and non-target organisms and the environment. Please note the reference protocols associated with each data point.

A) Molecular weight

Every chemical has a characteristic molecular "weight". In the case of polymers (long repeating chains of molecules), an average molecular weight is given.

Resources

OECD Guidelines for the Testing of Chemicals

B) Physical State

This indicates whether a substance or product is solid, liquid or gaseous at room temperature.

C) Appearance

This describes the physical form of the product. Examples include granular solid, gelatinous liquid or fine powder.

D) Particle Size

If the substance is solid, please describe the average particle size. Give the range of particle sizes and their proportion of distribution. This property may be a significant factor in determining the physical distribution and biological uptake of a substance.

Resources

OECD Guidelines for the Testing of Chemicals

ASTM International

E) Colour and Odour

Include this description for each ingredient as well as for the formulated product.

F) Olfactory Detection Limit

This is the minimum concentration (e.g., parts per million by volume in air), at which one can identify the substance by smell. This is important in determining whether or not smell can serve as an appropriate warning of the presence of the substance.

G) Density or Specific Gravity

Density of solids is measured as mass per volume (i.e., g/mL). For liquids, specific gravity compares density to that of water, which is assigned a value of 1 (i.e., a substance with a specific gravity of 2.0 is twice as dense as water). Specific gravity has no units and is measured at temperatures of 15.6, 20 and 25°C. These measurements are used in several ways, such as when predicting the behaviour of a substance in the environment and accounting for the inclusion concentrations in feed.

Resources

OECD Guidelines for the Testing of Chemicals

H) Refractive Index

As light passes from one medium to another the speed at which it moves changes. Changing the speed of light may cause it to "bend" or refract. This property varies from one substance to another and is sometimes useful in identifying substances through a simple accurate technique.

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I) Melting Point

This is the temperature (at a given pressure, 101.325 kPa: 1 atm) at which a solid becomes a liquid. In cases where the substance undergoes a chemical reaction (e.g., degradation, decomposition, rearrangement) other than melting, then the temperature at which the reaction occurs must be reported. As alternatives to the melting point, a pour point, softening point or sublimation point could be provided when appropriate.

Resources

OECD Guidelines for the Testing of Chemicals

J) Boiling Point

This is the temperature (at a given pressure, 101.325 kPa: 1 atm) at which a liquid becomes a gas. In cases where the substance undergoes a chemical reaction (e.g., degradation, decomposition, rearrangement) other than boiling, the temperature at which the reaction occurs must be reported.

Resources

OECD Guidelines for the Testing of Chemicals

K) Flash Point

Flash point is the minimum temperature at which a liquid can ignite upon contact with a flame or spark in the presence of oxygen. This has obvious implications for worker safety during handling and storage.

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L) Auto-ignition Point

The auto-ignition point is the minimum temperature at which a flammable liquid will ignite spontaneously without an ignition source.

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M) Vapour Pressure

This is a measurement of a liquid's ability to evaporate, or give off vapours at specific temperatures. It is usually expressed in millimetres of mercury (mmHg), and is a crucial indicator of the behaviour of a liquid product, indicating whether it will tend to escape to the atmosphere or remain in soil. The vapour pressure is not required if the chemical has a standard boiling point less than 0°C.

Resources

OECD Guidelines for the Testing of Chemicals

N) Vapour Density

The vapour density is a comparison between the mass of a gas and that of dry air which is assigned a value of 1, (for example, the substance with a vapour density of 2.0 is twice as dense as air). Less dense gases tend to rise and are quickly transported throughout the environment; heavier gases remain closer to the ground and dissipate less rapidly.

O) Henry's Law Constant

This is a measure of the solubility of a gas in liquid. It is indicative of a substance's tendency to move from water to air or vice-versa.

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P) Potential of hydrogen (pH)

This property indicates whether a substance is acidic, neutral or basic and quantifies the strength of acidity or alkalinity. The pH of a substance can have a major effect on its interactions with living organisms, such as the degree to which it is absorbed or taken up as well as contact exposure for workers.

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Q) Solubility

i) Water

Solubility describes the amount a substance will dissolve in water at a given temperature. Since many chemicals dissolve in water to a significant degree, it is often the route by which chemicals are taken up by organisms. It is also used in predictions of environmental fate.

Resources

OECD Guidelines for the Testing of Chemicals

ii) Other Solvents

Solubility describes the amount a substance will dissolve into solvents other than water. This provides a guide for choosing a solvent to extract a substance from complex matrices (i.e., tissue or soil).

Resources

OECD Guidelines for the Testing of Chemicals

R) Octanol-Water Partition Coefficient

This parameter measures the tendency of a substance to partition, either into organic solvents or water. It is used in predicting whether a substance may accumulate in the fatty tissues of an organism and predicts its tendency to adhere to soil particles.

Resources

OECD Guidelines for the Testing of Chemicals

S) Dissociation Constant

Dissociation is a specific type of chemical decomposition in which a molecule breaks up into charged particles called ions. Ions are often involved in further chemical reactions and may be absorbed or distributed at a different rate. For given conditions, the rate of dissociation (dissociation constant, or Kd) is constant.

Resources

OECD Guidelines for the Testing of Chemicals

T) Incompatibility

Dissociation is a specific type of chemical decomposition in which a molecule breaks up into charged particles called ions. Some substances react with other substances. If the formulated product or any of its ingredients come in contact with certain substances, this must be reported.

U) Polymerization

Some substances may spontaneously polymerize (form long molecular chains) under certain conditions. Many of these reactions can produce dangerous or explosive amounts of heat. Any such substances contained in the formulated product must be identified along with a description of conditions under which spontaneous polymerization is known to occur.

5.0 Mammalian toxicological data

In order to assess potential harmful effects of a substance to mammals, data from laboratory tests is necessary. Test animals must be dosed using the same route or routes of exposure that are anticipated to be the most significant route or routes for livestock and potential human exposure (e.g., oral, dermal or inhalation). Please note that the most significant route of exposure to a substance for the general human population may be different from exposures for workers in an occupational setting.

The type of toxicity data generally considered is outlined below.

A) Toxicokinetic Studies (Absorption, Distribution, Metabolism and Elimination)

For each route by which a substance can be taken into the body (oral, dermal, respiratory, etc.), absorption tests determine how extensively and how rapidly it can be absorbed.

Distribution, metabolism and elimination data describe the fate of a substance once it is absorbed into the body. Questions answered by this information include: What is the metabolic fate of the substance? Does it accumulate? How is it broken down or transformed? What metabolites are formed? By what route and how quickly are the substances eliminated? This information provides an indication of an organism's ability to tolerate short- or long-term exposure, either at high or low concentrations. It is also useful in designing toxicity tests (e.g., dose selection), and the extrapolation of data from animal models to human exposure.

Resources

OECD Guidelines for the Testing of Chemicals

B) Acute Toxicity Studies

Acute exposure tests examine the effects of single exposures to high concentrations of a substance. Exposures in these tests are typically 24 hours or less, with effects being monitored for up to two weeks. This is the first step in establishing the crucial relationship between the dose and the response, for ranking substances according to their relative acute toxicity, and for classification and precautionary label statements. Acute toxicity data is used to obtain preliminary information on specific toxic effects of substances and how these may be produced (mode of action). Some specialized acute tests are described below.

Resources

OECD Guidelines for the Testing of Chemicals

United States Code of Federal Regulations

i) Acute Median Lethality Studies

The concentration of a substance, when administered once to a group of animals for a short time, will cause death in half of the animals. This is expressed as an LD50 (lethal dose, in mg/kg of body weight) or the LC50 (lethal concentration, in parts per million).

Resources

OECD Guidelines for the Testing of Chemicals

ii) Skin/Eye Irritation Studies

This determines if a substance has the potential to cause irritation or cell death (necrosis) when in contact with the skin or eyes.

New methods must be assessed by the CFIA to determine whether they provide sufficient information, therefore, we advise that you contact the Animal Feed Division regarding the validity of methods that are alternatives to animal testing.

Resources

OECD Guidelines for the Testing of Chemicals

United States Code of Federal Regulations

iii) Skin Sensitization Studies

This evaluates the potential of a substance being able to "sensitize" organisms. Organisms may become more sensitive to a substance after an initial exposure and develop allergic reactions in subsequent exposures.

Properly conducted patch tests (positive or negative response) may be an acceptable alternative to animal testing. The concentration of a substance to exposed subjects will be a critical factor regarding the acceptability of the patch tests. In addition, information for the assessment of skin or ocular irritation may be obtained from Quantitative Structure-Activity Relationships (QSARs), with adequate scientific justification provided by the notifier regarding the validation and applicability domain of the model.

Resources

OECD Guidelines for the Testing of Chemicals

United States Code of Federal Regulations

C) Mutagenicity/Genotoxicity Studies

Mutations to DNA may lead to cancer or gene-based malformations in offspring. Screening tests are used in determining a substance's potential for causing genetic mutations. At least two types of tests are performed, conventionally one with bacteria and one with mammalian cell cultures. It is also necessary to perform tests with and without metabolic activation, that is, to determine if interactions with metabolic processes in the body results in a substance being mutagenic.

Resources

OECD Guidelines for the Testing of Chemicals

Health Canada

United States Code of Federal Regulations

D) Short-Term Toxicity Studies

Short-term toxicity studies involve repeated exposure to substances over a longer period of time (i.e. 14-90 days). They are useful for detecting long-term adverse health effects, for establishing a threshold level at which no effects are observed, for establishing possible cumulative effects of exposure, species variation, and suggesting appropriate conditions for chronic tests, if deemed necessary. A 90-day oral study is typical, and inhalation or dermal studies may be appropriate, depending on typical human exposure parameters.

Resources

OECD Guidelines for the Testing of Chemicals

E) Chronic Toxicity Studies

In some cases, chronic studies may be necessary to evaluate the safety of a substance over a longer period of time, such as the lifetime of the animal (e.g. 120 days to 1-2 years). These studies are practical for detecting subtle, adverse health effects, such as carcinogenesis and reproductive effects. Some of the chronic studies are outlined in the following sections.

Resources

OECD Guidelines for the Testing of Chemicals

F) Teratogenicity/Developmental Toxicity Studies

The teratogenicity studies investigate adverse effects induced during development from conception until birth.

The developmental toxicity tests examine adverse effects during the lifetime of an organism prior to conception, during pre-natal development or until sexual maturity, resulting from exposure of either parent to a substance.

Resources

OECD Guidelines for the Testing of Chemicals

United States Code of Federal Regulations

G) Reproductive Toxicity

These studies investigate the adverse effects of substances on male or female reproductive systems and thus their reproductive capacity from mating through to lactation. Typically, they are multi-generational studies.

Resources

OECD Guidelines for the Testing of Chemicals

United States Code of Federal Regulations

H) Neurotoxicity

These studies investigate the adverse effects of substances on the nervous system, (central nervous system and peripheral nervous system).

Resources

OECD Guidelines for the Testing of Chemicals

I) Carcinogenicity

When indicated by earlier tests or other data, a substance will be examined for its ability to induce cancer (tumours) in animals. Tests are typically conducted over a major portion of the animal's life span, often combined with chronic toxicity tests.

Resources

OECD Guidelines for the Testing of Chemicals

United States Code of Federal Regulations

J) Estrogenicity Studies

Resources

OECD Guidelines for the Testing of Chemicals

K) Epidemiological Studies

This type of study contains a compilation and analysis of information on humans who have been occupationally or accidentally exposed to a substance.

6.0 Suggested maximum residue limit (MRL) or tolerance

A Maximum Residue Limit (MRL) governs the concentration of a chemical that may accumulate in tissues of livestock without causing harm to the animal, or to humans that consume their products. This limit should be based upon an evaluation of information such as mammalian toxicity data, dietary intake estimates, and livestock metabolism data.

7.0 Human exposure data and exposure estimation

The following information is important in assessing the degree of exposure to workers and users of particular substances.

A) Major Routes of Exposure

Human exposure to a substance depends, in part, on how it can be taken up by the body, that is to say via the respiratory route, skin absorption, oral ingestion, etc. Based on the ingredients, its formulation, and its methods of use, please describe how users might be exposed.

B) Amount of Product Handled by Workers and Consumers

Give the total quantity of product that would typically be used by workers or those who have consumed the product on a daily basis (e.g. 50 litres/day).

C) Frequency and Duration of Exposure

State how often and with what frequency the product will normally be used. Indicate the duration of exposure for each use period.

D) Exposure Concentrations

State the concentration of the product when transported, stored and used. Please describe any intermediate preparation steps, such as mixing or dilution.

E) Exposure Studies

Include all data on human exposure, uptake into the body, and medical studies of workers who have been exposed to the product over long periods of time. In some cases exposure studies may have to be conducted to establish how much absorption and distribution takes place with intended conditions of use.

8.0 Livestock metabolic fate and residue studies

Mammalian toxicity data derived from tests on mice and rats offer limited information on the risk of a particular substance to livestock species. The following data requirements involve direct testing with livestock animals.

Resources

A) Toxicokinetic Studies in Livestock Species

These tests describe what happens to a substance once it is ingested by livestock. The objectives are to establish the rate and mode(s) of absorption into systemic circulation, the distribution in the body, the rate of metabolism, possible metabolic pathways and all metabolites. Metabolic parameters to be evaluated include the rate and routes of elimination and the biological half life (t½). These data requirements should also include statistically significant analytical techniques for recovery and detection limits in all tissues evaluated.

Resources

OECD Guidelines for the Testing of Chemicals

B) Residue Studies for the Parent Compound and Metabolites

Residues in livestock tissue have the potential to enter the human food chain. Thus, it is important to determine how much of a substance actually remains in exposed animals and in which tissues it is found. This data requirement should also include statistically significant analytical techniques for recovery and detection limits in animal tissues.

Resources

OECD Guidelines for the Testing of Chemicals

9.0 Environmental fate and effects

The following data requirements, in combination with the physicochemical data, form the basis for predicting or determining which environmental compartments will be exposed to a substance following its release (or the release of its metabolites). It will also assist in the determination of how non-target organisms may be affected.

A) Vapour Pressure and Volatilization

This is a measure of a liquid's ability to evaporate or give off vapours at specific temperatures. Usually expressed in millimetres of mercury (mmHg), it is a crucial indicator of the behaviour of a liquid product, for example whether it will escape to the atmosphere or remain in soil.

Resources

OECD Guidelines for the Testing of Chemicals

United States Code of Federal Regulations

B) Hydrolysis

One of the main ways in which substances break down in the environment is by hydrolysis. The extent to which this reaction takes place under normal conditions, and the products of this type of reaction should be described. Please note that not all substances undergo hydrolysis reactions.

Resources

OECD Guidelines for the Testing of Chemicals

C) Photodegradation

Interaction with light is another important way in which substances are degraded in the environment. Photodegradation tests determine the potential for light-induced changes, as well as possible transformation products formed following this type of reaction.

Resources

D) Adsorption-Desorption

The ability of a substance to adsorb ("attached") or to desorb (detach) from other particles or molecules affects how quickly it may move through a particular environmental compartment. Soil is a major adsorption medium for many substances.

Adsorption and desorption are often mediated by pH. Please account for the pH of the soil or other media.

Resources

OECD Guidelines for the Testing of Chemicals

United States Code of Federal Regulations

AOAC Methods of Analysis

E) Simple Biodegradation Tests

Resources

OECD Guidelines for the Testing of Chemicals

F) Biotransformation in Soil

Some substances undergo changes as a result of transformation by micro-organisms (i.e., microbial activities). These tests determine the potential for biodegradation, bioactivation or biotransformation taking place. Data for both aerobic and anaerobic conditions (in the presence or absence of oxygen) are useful because different micro-organisms are involved in biotransformation processes.

Resources

OECD Guidelines for the Testing of Chemicals

AOAC Methods of Analysis

G) Biotransformation in Aquatic Systems

In addition to soil biotransformation, substances may be transformed by the microbes in the aquatic environment. Data for both aerobic and anaerobic conditions are useful because different micro-organisms are involved in biotransformation processes.

Resources

OECD Guidelines for the Testing of Chemicals

AOAC Methods of Analysis

H) Biochemical Oxygen Demand

Oxygen demand is the quantity of oxygen required by micro-organisms to oxidize organic compounds in water. Results are measured as dissolved oxygen (DO) in mg of oxygen per litre (mg O2/L) or mg of oxygen per gram (mg O2/g) of compound. Organic contaminants can impair water quality by reducing oxygen levels which in turn can adversely affect aquatic organisms.

Resources

OECD Guidelines for the Testing of Chemicals

I) Toxicity to Aquatic Organisms

It is important to establish the adverse effects of substances on aquatic organisms. Often the mechanisms of these effects are unique to aquatic environments and toxicity cannot be predicted using laboratory mammals. Also, the repercussions of disturbing organisms at the bottom of the food chain can be of great concern to the entire ecosystem.

Resources

OECD Guidelines for the Testing of Chemicals

United States Code of Federal Regulations

J) Toxicity to Soil Organisms

The adverse effects of a substance on soil organisms are important because the long-term health of agricultural soils can be compromised by disturbing populations and communities.

Resources

OECD Guidelines for the Testing of Chemicals

ISO documents

K) Toxicity to Birds

The adverse effect of substances on birds can be important. Birds may be more susceptible to certain substances than laboratory animals, and therefore specific tests may be required. The difference in tolerance is partly due to differing environmental stresses and conditions as well as to metabolic and behavioural differences.

Resources

OECD Guidelines for the Testing of Chemicals

L) Toxicity to Wildlife

These toxicity tests are valuable for indicating the adverse effects a substance will have on animals in real environmental conditions. In the case of mammalian wildlife, livestock data may be suffucient.

Resources

OECD Guidelines for the Testing of Chemicals

ASTM International

2.6 Guidelines for the assessment of novel feeds: Plant sources

1.0 Introduction

1.1 Scope

These guidelines (Section 2.6 of Regulatory Guidance: Feed Registration Procedures and Labelling Standards) apply to novel feeds from plants, or parts or products thereof, which have not previously been used as livestock feed in Canada, i.e. not listed in Schedules IV or V of the Feeds Regulations, and/or have a novel trait making them not substantially equivalent to an approved product already on the market in Canada.

Prior to the release of a novel feed (i.e., the manufacture, sale, import or use as livestock feed) authorization from the Animal Feed Division, CFIA, is required.

These guidelines are a guide for the preparation of an application for the authorization of the release of a novel feed. It includes criteria that will be considered in the assessment of safety and efficacy of a novel feed. This directive is not intended to explicitly define all of the data that could be required in the course of the assessment.

More information on novel feeds, application procedures and Animal Feed Division activities is available on the CFIA Web Site.

These guidelines have been prepared to provide guidance regarding the submission of an application for the authorization of the release of a novel feed, as may be required under the Feeds Regulations. Please be aware that the information provided in these guidelines is not exhaustive and will be updated as appropriate to reflect current scientific knowledge. For further clarification, applicants are strongly recommended to consult with the CFIA's Animal Feed Division. For all purposes of interpreting and applying the law, applicants are invited to consult the official versions of the relevant Acts and Regulations.

1.2 Legal Authorities

The manufacture, sale and import of livestock feeds are regulated in Canada under the Feeds Act and Regulations. All feeds must be safe, to livestock; to humans (via the potential transfer of residues into human food, i.e., meat, milk and eggs, and via worker/bystander exposure); and to the environment. Feeds must also be shown to be effective for their intended purpose. Approved feed ingredients are listed and defined in Schedules IV and V of the Feeds Regulations, with appropriate guarantees, standards, and label requirements. All imported feeds must meet the same standards as domestic feeds.

The legislative authorities under which the CFIA regulates novel feeds are as follows:

For the regulation of novel feeds:

The Feeds Act, R.S. 1985 chapter F-9
The Feeds Regulations, 1983 F-9 SOR/83-593

For the collection of fees:

Canadian Food Inspection Agency Fees Notice, Canada Gazette, Part I (05/13/2000)

1.3 Regulation of Novel Feeds

The CFIA evaluates and regulates all feed ingredients, including novel feeds, in the same manner. Any feed ingredient that is new (i.e., is not already listed in Schedules IV or V of the Feeds Regulations), or has been modified such that it differs significantly from a conventional ingredient, is required to undergo a pre-market assessment and approval. The purpose of all feed assessments is the same: to ensure that the feed ingredient is safe (in terms of animal health, human health via residues in food and worker/by-stander exposure, and the environment) and effective for its intended purpose prior to marketing. The evaluation also ensures that the feed is accurately defined in the Feeds Regulations and is labelled appropriately for its safe, effective use and for consumer protection.

Novel feeds are considered to be those derived from an organism or organisms, or parts or products thereof that:

  1. are not approved as livestock feed in Canada (i.e. are not listed in Schedule IV or V of the Feeds Regulations), and/or
  2. contain a novel trait.

Schedules IV and V of the Feeds Regulations list feed ingredients approved for use in livestock feed in Canada. Schedule IV comprises a range of ingredients such as forages and roughages, energy feeds, protein sources, vitamins, minerals, fermentation products and other products, while Schedule V is restricted to flavouring ingredients.

A novel trait is a heritable characteristic of a feed that is not substantially equivalent in terms of its specific use and safety to a characteristic of a similar feed that is set out in Schedule IV or V of the Feeds Regulations. In other words, for novel feeds derived from plant sources, a novel trait is a heritable characteristic that is new to a plant species, or is an endogenous trait that has been modified such that it differs from conventional parameters for that plant species.

Note that for feeds with novel traits, it is the presence of the novel trait that triggers regulatory oversight under the Feeds Regulations, and not the method used to introduce the trait; hence the plant, rather than the process, is subject to regulatory oversight. Novel feeds may be created by such methods as traditional breeding, mutagenesis, cell fusion, or recombinant DNA techniques.

It is also important to note that the trigger for regulatory oversight is novelty, not risk. Any potential risk, in terms of safety and efficacy, is determined during the assessment of the product.

Proponents seeking guidance with respect to determining whether or not their product is a novel feed can refer to Appendix II of these guidelines or consult directly with the Animal Feed Division. Consultation with the Animal Feed Division is highly recommended for any proponents experiencing difficulty in determining novelty.

1.4 Assessment Principles

Feed assessments are conducted to ensure that feed ingredients are safe (in terms of animal health, human health via residues in food and worker/by-stander exposure, and the environment) and effective for their intended purposes prior to marketing. The Animal Feed Division conducts livestock feed assessments based on several internationally recognized principles.

  1. Assessments are conducted on a case-by-case basis.

    Novel feeds can vary greatly in terms of their individual characteristics. Thus the prescription of one set of data requirements to demonstrate efficacy and safety for all assessments would be impractical and onerous for proponents. Data requirements are, instead, determined on the basis of the characteristics of the product in question. This allows proponents to concentrate on meeting those data requirements relevant for their particular product.

  2. Familiarity

    The degree of scientific support required in a data package is based on the complexity of the product in question and the degree of familiarity associated with the product/trait. If the Animal Feed Division has a high degree of familiarity with respect to the characteristics of a particular product and/or trait, in terms of livestock feed use, the level of data required to demonstrate safety and efficacy may be reduced or more narrowly defined.

  3. Comparative Approach

    Novel feed assessments are done using a comparative approach. A comparison of a novel feed against products with a history of use as livestock feed highlights the similarities and differences that the novel feed has when compared to feeds already present in the marketplace.

    Comparisons of data made within the submitted data package should make use of an appropriate counterpart or counterparts. Appropriate counterparts have a history of livestock feed use within Canada. For feeds with novel traits, proponents should typically make comparisons using the unmodified counterpart. In other words, proponents should use conventional parental varieties and/or, in some cases for plants derived using recombinant DNA techniques, negative segregants when making comparisons.

  4. Valid Scientific Rationale

    Provision of a valid scientific rationale (supported by appropriate data or literature references) may negate requirements for certain studies detailed in these guidelines or allow bridging to other valid scientific studies.

  5. Weight of Evidence

    A weight of evidence approach is used when performing assessments. This means that the sum of the overall data submitted provides the context for determining whether or not a novel feed should be authorized.

  6. Feed Definitions & Labelling

    A component of the approval process for novel feeds is to ensure that they are accurately defined within the Feeds Regulations and are labelled appropriately for safe, effective use and for consumer protection. In some instances a novel feed may be considered as meeting an existing definition, while in other instances an existing definition may need to be modified, or a new definition may be created.

    Additional principles applying to feeds with novel traits:

  7. Substantial Equivalence

    Substantial equivalence is based on the comparison of properties between the modified plant from which a feed with a novel trait is derived and an appropriate counterpart. Taking into consideration both intended and unintended effects, similarities and differences between the modified plant and its counterpart can be identified. Those aspects in the modified plant which are determined to be equivalent to the counterpart are accepted and the assessment then focuses on the differences. The concept of substantial equivalence is endorsed by international groups such as the OECD and FAO/WHO.

    In the feed assessment, molecular, nutritional and toxicological data for the novel feed are compared to those of an appropriate counterpart. The identification and subsequent determination of levels of key nutrients, antinutrients and endogenous toxins and allergens is an important aspect of the determination of substantial equivalence. The OECD Consensus documents on individual crops may be of assistance as a reference to aid in the determination of the key elements to be examined.

  8. Unintended Effects

    The assessment process considers the likelihood that unintended effects may be present in the modified plant and that these unintended effects may impact efficacy and/or safety of feed derived from the plant.

1.5 Determining Data Requirements

Section 2-8 of these guidelines outline the categories of data requirements that proponents should consider when compiling a novel feed submission. Proponents should address each section, but the manner in which the requirements can be satisfied will vary. In some instances collection and analysis of raw data is necessary, while in other instances providing a valid scientific rationale supported by existing scientific publications is sufficient.

Applicants are encouraged to consult with the Animal Feed Division in the early stages of the development of a novel feed for clarification on the specific information that is required to conduct a safety and efficacy assessment. Please note that additional regulatory requirements exist if a proponent wishes to make claims associated with their feed once it is authorized. Guidance for feed claims can be found in Chapter 2, Section 2.2, of Regulatory Guidance: Feed Registration Procedures and Labelling Standards.

1.6 Data Quality

Data provided as part of an application must be of a standard that would be acceptable for publication in a peer-reviewed scientific journal. Studies must be designed and conducted in accordance with sound scientific concepts and principles and, where appropriate, Good Laboratory Practices. All test substances used in the generation of data should be representative of the final feed form. Controls should also be appropriate to meet the objective(s) of the study. Studies must be carried out by qualified research personnel using validated methods. Complete descriptions of the methods used to generate submitted data must be provided, including the sensitivity and quality control/quality assurance procedures. When applicable, data must be analyzed using appropriate statistical analysis techniques. Primary data must be made available upon request. All parts of the application must be fully legible, with particular attention paid to analytical gels and blots (e.g., SDS-PAGE gels, Northern blots, Southern blots, and Western blots). Any scientific references used to support the data package should be provided with the application.

A reviewer's checklist for analytical techniques, developed in collaboration with Health Canada and the United States Department of Agriculture's Animal and Plant Health Inspection Services, providing guidance on the preparation of quality data.

In light of the rapid ongoing adoption of whole genome sequencing (WGS) in pre-market submission packages, a guidance document was developed describing the principles and good practices that proponents should consider in organizing and presenting their data. Proponents are encouraged to consult with "Guidance for submitting whole genome sequencing (WGS) data to support the pre-market assessment of novel foods, novel feeds, and plants with novel traits" when submitting WGS-based analysis in their submission package.

1.7 Presentation of Information

Proponents should organize their submissions in a manner similar to that of a scientific paper suitable for publication. A typical submission format will consist of:

Note that within the concluding statements proponents should clearly indicate how the results relate to product safety and/or efficacy with respect to use as livestock feed.

1.8 Procedures for Submitting an Application

Applicants are strongly encouraged to apply for authorization of the release of the novel feed well in advance of the anticipated time of commercialization. Applicants are required to follow the procedures for submitting an application as outlined in Chapter 1 of Regulatory Guidance: Feed Registration Procedures and Labelling Standards.

Please note that these guidelines are not intended to explicitly define all of the data that might be required in the course of the assessment. Additional information may be requested by the Animal Feed Division on a case by case basis. Applicants are encouraged to consult with the Animal Feed Division in the early stages of the development of a novel feed for clarification on the specific information that is required by the Animal Feed Division to conduct an assessment.

Applicants must submit a hard copy version of the application and are encouraged to also submit an electronic (compact disc) copy of the application.

1.9 Harmonization With Other Regulatory Parties

Where a novel feed is derived from a plant that could reasonably be expected to be released into the Canadian environment and/or used as human food, the Plant Biosafety Office, CFIA, and/or Novel Foods Section, Health Canada may also require applications for authorization of the Plant with a Novel Trait (PNT) and/or novel food, respectively. It is the responsibility of the applicant, and not of the Animal Feed Division, to contact these offices and determine if an application is required.

Applicants seeking authorization for the release of a novel feed in Canada are encouraged to seek authorization for their product in the United States and Canada simultaneously, if the product will be regulated in the United States. Obtaining such authorizations should minimize the movement of unauthorized material across the border.

When a novel feed is derived from a plant that is considered to also be a PNT and/or a source of a novel food, authorizations for use as a novel feed, novel food and environmental release will be granted in a harmonized fashion. This will minimize the potential for unapproved products to enter the Canadian environment or food or feed supplies. Proponents and importers should be aware that, where applicable, approvals are required from all three regulatory parties prior to environmental release, food use or feed use. Note that harmonization of authorizations is only required when a product triggers more than one type of assessment. In other words, novel feeds that are not novel foods or PNTs do not require harmonized authorizations.

Where plant-derived products are intended for exclusive use as food, or feed or for plant molecular farming, consultations among regulatory authorities will still be required to assess any potential risks, and management thereof, associated with release of the product into a commodity stream for which it is not intended.

Please note that once the assessments have been completed, the applicant is notified in writing by the CFIA and Health Canada (in separate letters) regarding their respective decisions regarding the application.

1.10 Confidential Information

Information provided to the CFIA by applicants for the purposes of obtaining authorization for the release of novel feeds can only be accessed by authorized personnel and is protected under the federal Access to Information Act, Section 20.

All third party requests for information are subject to the federal Access to Information and Privacy (ATIP) Acts. For further information, please consult with CFIA's ATIP service, at 613-773-2342.

2.0 Characterization of the plant source

Sufficient data must be submitted to characterize the novel feed, and the plant from which a novel feed is derived, to permit comparison with appropriate counterparts.

Most of the questions regarding product characterization can be addressed by data generated in the developmental stage.

2.1 Source, Host and Donor Organisms

For feed derived from a plant source with no history of use as feed, characterization of the plant source is essential to assess the efficacy and safety of the novel feed.

For feeds with novel traits, information regarding the host and any donor organisms used in the development of a modified plant can, in part, lead to a better understanding of what needs to be assessed. In some cases, characterizing these organisms is quite straightforward, e.g. for those that are conventional crop species. Conversely, donor organisms that have no history of use as livestock feed, or those known to contain toxins, allergens or antinutrients, will require a more detailed characterization.

Required information for applicable source, host and donor organisms includes, but is not restricted to, the following:

  1. Common or usual names; scientific name and taxonomic classification.
  2. A critical assessment of the ability of the organism to produce/contain potentially toxic compounds (e.g., heavy metals, pesticides), anti-nutritional compounds and endogenous allergens.
  3. Available toxicology data for the organism.
  4. Information on history of use of the organism, or any related organisms, in livestock feed.

2.2 Plant Characterization

Information regarding the characteristics and use of a plant as a feed source are core components of an application.

Provide the following information:

  1. All designations for the plant used in the application.
  2. A description of the intended feed use of the plant.
  3. List of other government agencies, in Canada or abroad, which were notified of the development of the novel feed, and description of the purpose of the notification; and the countries in which the novel feed has received regulatory approval.

    For a feed with a novel trait also provide:

  4. A description of the novel trait.
  5. A description of the intended purpose of the novel trait.
  6. A description of how the proponent analyzed the host plant to determine if any anticipated secondary effects or unintended effects were present.

    This assessment takes into consideration the possible presence of unintended effects in the modified plant; whether present as a result of the modification process or due to the modification itself. Overall phenotypic characterization of the modified plant can be used as evidence to show a lack of unintended effects or to identify any such effects.

    There are two ways in which plants are often characterized:

    1. compositional profile comparisons with an appropriate counterpart, and
    2. agronomic/phenotypic parameter comparisons with an appropriate counterpart.

    Note that proponents are not required to provide the Animal Feed Division with agronomic/phenotypic data if this information has already been provided to the CFIA as part of a PNT assessment.

    To satisfy this requirement, proponents may refer to data they have provided to address Section 3 of these guidelines (compositional data) and data they have provided as part of the evaluation of environmental safety of the modified plant.

  7. A characterization and evaluation of any secondary effects and unintended effects that are identified in the host plant (in terms of the safety and efficacy of the derived feed).

2.3 Modification/Development History

For feeds with novel traits, provide the following information:

  1. Describe the method of trait introduction/selection.

    Different methods of trait introduction have different characteristics associated with them with respect to both intended and unintended effects. Knowledge of the method used to introduce the modification can, in part, lead to a better understanding of what needs to be assessed in the modified plant.

  2. An outline of the pedigree of the modified plant. Please indicate which generations were used to produce supporting data.
  3. Describe the selection criteria that were employed at both the initial stages of development and during the subsequent breeding process.
  4. Evidence to support the inheritance and stability of each genetic modification in the plant.
    • For plants which are either male- or female-fertile, or both, data that demonstrates the pattern and stability of inheritance and the expression of the new traits is required. If the new trait cannot be directly measured by an assay, it may be necessary to examine a) the inheritance of the DNA insert directly, and b) RNA expression.
    • For plants which are either infertile, or for which it is difficult to produce seed (such as vegetatively propagated male-sterile potatoes), data to demonstrate that the trait is stably maintained and expressed during vegetative propagation over a number of cycles appropriate to the crop are required.
  5. In the case of an allopolyploid plant, name the parental genome in which the genetic modification occurred.

2.4 The Novel Trait(s)

For feeds with novel traits, provide the following information in regards to the novel trait and any associated newly expressed material that is either exogenous or modified native material:

  1. A description of the history of use of the novel trait.
  2. Characterize how the novel trait operates.

    Information regarding how a novel trait operates can, in part, lead to a better understanding of what needs to be assessed. Define the genetic basis of the trait (e.g. the inserted gene, mutation) and describe the downstream effect(s) of this genetic basis that result in the novel trait. For products of rDNA, consider all genetic material potentially transferred to plant host genome and characterize DNA shown to be transferred (see Appendix I).

  3. Detailed information on the gene product(s) including their activity, breakdown products, byproducts and their affected metabolic pathways (including altered accumulation and storage patterns), resulting from the novel trait.
  4. Indicate if gene expression is constitutive or inducible, with a description of the inducing agents. Provide information demonstrating that the trait is expressed in the appropriate tissues in a manner and at levels that are consistent with the associated regulatory sequences driving the expression.
  5. Characterize the expression level of the gene product in all plant parts that may be fed to livestock. A determination of the levels of the metabolites of the gene product may be required.
    • If the protein concentration is below the limit of detection, mRNA data may be substituted.
    • For virus resistant plants where the transgenes are derived from a viral genome, transgene RNA levels in tissues consistent with the associated regulatory regions driving expression of the transgene are required in addition to transgene protein analysis.
    • For plants modified to specifically alter the accumulation of a specific mRNA or protein, data on the level of the target protein only may be provided. If the target protein levels are below levels of detection, mRNA levels may be provided.
  6. The likelihood of exposure to livestock, humans and the environment, including the estimated level and most likely route of exposure to the novel products, breakdown products, and by products.
  7. The potential for adverse health effects to humans and livestock, including exposure to toxins and anti-nutritional factors, as well as irritation and allergenicity to humans (in terms of bystander/occupational exposure).
  8. The potential of carry over of any novel product to manure and the potential impact of this carry over.

2.5 Marker Genes

Any transformation markers used in the development of a plant must be identified and characterized as per the novel trait (Section 2.4 of these guidelines). Any secondary effects of these markers on the biochemistry, physiology and secondary metabolism of the host plant, must also be characterized and discussed with respect to consequences regarding efficacy and safety.

Because antimicrobials are routinely mixed with livestock feed, a determination of the potential for their inactivation during storage as a result of the activity of any expressed antimicrobial resistance marker gene should be conducted.

The use of marker genes that confer resistance to clinically important antimicrobials should be avoided.

3.0 Nutritional data

Novel feed ingredients derived from the plant should be described, including information on processing and estimated feeding rates of the ingredient and by-products.

Nutrient and antinutrient composition of the major plant components and byproducts used in livestock feed must be considered. For example, composition of whole plant corn or corn silage in addition to grain should be determined. Consideration must also be given to by-product streams in which nutrients or antinutrients are concentrated.

A comparison of the composition of the novel feed ingredient to that of the same feed ingredient derived from an appropriate counterpart must be undertaken. These comparisons should be based on analysis of representative samples derived from appropriate growing areas over one or more growing season(s). Comparison to literature ranges for nutritional composition of nearest genetic equivalents or counterparts may be an acceptable alternative. Data should demonstrate the uniformity or variability of the composition of the final product and include the analysis/characterization of the gene products or any metabolites that may have been altered (e.g., in the case of proteins, post translational modifications are of interest).

Scientific investigations presented in support of nutritional evaluation must be: designed to facilitate statistical analysis; analyzed by appropriate statistical methods; and, for those feeds which are able to grow in Canada, conducted under growing/environmental conditions similar to those expected to occur in Canada. As a minimum, the experimental design should include three locations, two treatments (novel feed vs. control), and three replicates. Representative samples should not be pooled for analysis. Nutrient and antinutrient analysis should be done in duplicate, according to accepted AOAC International or other recognized methods.

The raw data and printouts of statistical analysis must accompany the application. Statistical differences in composition should be addressed in terms of biological significance.

3.1 Nutrient Composition

Provide, at a minimum:

  1. Crude protein
  2. Crude fat and ash
  3. Crude fibre or acid detergent fibre (ADF) and neutral detergent fibre (NDF).
  4. Levels of naturally occurring or adventitious antinutritional factors which could reasonably be expected to be present (e.g., phytates, trypsin inhibitors, alkaloids, pigments, etc.)

    Based on the preceding evaluation and the intended use of the feed, the following information may also be required:

  5. True protein, non-protein nitrogenous material, and amino acids
  6. Quantitative and qualitative composition of total lipids, i.e., saponifiable and non-saponifiable components, and fatty acids
  7. Composition of the carbohydrate fraction, e.g., sugars, tannins, non-starch polysaccharides and lignins
  8. Vitamins
  9. Minerals

For feeds with novel traits, the OECD consensus documents, which detail the key nutrients and antinutrients to be analysed in various crop species, should be consulted for guidance.

Unusual or unanticipated results in nutrient or antinutrient analysis must be investigated further.

In the case of feeds with novel traits which have been intentionally modified in terms of nutritional composition (e.g., altered amino acid or fatty acid profile) or functionality (e.g., digestibility, bioavailability), nutrient and antinutrient analysis and comparison with the appropriate controls should be conducted as described above. Data to support any claims associated with the modification must also be provided. A discussion of the potential consequences to safety and efficacy of the intentional nutritional modification should be provided.

4.0 Dietary exposure

Complete details should be supplied on the predicted amount of the novel feed in the complete livestock diet. Estimates of dietary exposure to the novel feed may play a key role in determining the data required for a safety and efficacy assessment. In addition, data specific to animal age and species with respect to the intended use of the novel feed may be necessary. Feeds that are present at low levels in a complete diet may be of less concern than those intended to be significant components of the diet. Proponents should also discuss any impact the novel feed may have on conventional feed ration formulation.

For feeds with novel traits, the predicted amount of the feed in the complete livestock diet is typically based on the traditional use of the unmodified counterpart. Estimates of dietary exposure should be conservative in that they should consider situations whereby 100% replacement of the unmodified counterpart occurs.

If a feed with a novel trait will, or should, be used in a way that will result in dietary exposure that is different from that of the unmodified counterpart, this should be noted and the significance of the changed pattern of use discussed.

If a feed with a novel trait has intentional changes in a compositional value, discuss how this change will affect animal health and production.

5.0 Toxicology data

The toxicity of the host and donor organisms and/or the introduced novel traits to livestock through feed intake and humans through ingestion of livestock-derived food products (i.e., meat, milk, and eggs) and bystander/occupational exposure should be considered.

5.1 Endogenous Toxins

  1. For novel feeds that are new feed ingredients, provide data on the level of endogenous toxins known to be present in the plant from which the novel feed is derived and compare these with levels found in approved feed ingredients.

    For feeds with novel traits, provide a comparison of the levels of endogenous toxins known to be present in host or donor organisms between the modified plant and the unmodified counterpart.

5.2 Newly Expressed Material

Provide the following information for feeds with novel traits in regards to newly expressed material, which is either introduced exogenous material or modified native material:

  1. A comparison of the amino acid sequence of the newly expressed protein to amino acid sequences of known toxins to determine any potential homology. Specify the database(s) used to carry out the search.
  2. Description of any similarity of newly expressed material resulting from the modification to products having a history of use.
  3. The digestive stability of newly expressed protein(s).
  4. The heat stability of newly expressed protein(s).
  5. If concerns arise from assessment of the data submitted, additional toxicity studies may be required.

If bacterially produced protein is used in place of plant produced protein in studies of the potential toxicity of the novel protein, biochemical, structural and functional equivalency of the two proteins must be demonstrated.

Please refer to Chapter 2, Sections 2.3 to 2.5, of Regulatory Guidance: Feed Registration Procedures and Labelling Standards for further information on traditional toxicity study requirements for feeds.

6.0 Allergenicity data

The allergenicity potential of the host and donor organisms and/or the introduced novel traits to humans through ingestion of livestock-derived food products (i.e., meat, milk, and eggs) and bystander/occupational exposure should be considered.

6.1 Endogenous Allergens

  1. For novel feeds that are new feed ingredients, provide data on the level of allergens known to be present in the plant from which the novel feed is derived and compare these with levels found in approved feed ingredients.

    For feeds with novel traits, provide a comparison of the levels of endogenous allergens known to be present in host, parental lines, or donor organisms between the modified plant and an appropriate counterpart.

6.2 Newly Expressed Proteins

At present there is no definitive test that can be relied upon to predict allergic response to a newly expressed protein. A case-by-case, step-wise approach (such as the FAO/WHO 2000 or 2001 decision tree) should be applied in the assessment of possible allergenicity of newly expressed proteins.

Provide the following information for feeds with novel traits in regards to newly expressed proteins which are either introduced exogenous material or modified native material:

  1. A comparison of the amino acid sequence of the newly expressed protein to amino acid sequences of known allergens. The size of the contiguous amino acid search should be based on scientifically justified rationale to minimize the potential for false negative or false positive results. Validated search and evaluation procedures should be used. Specify the database(s) used to carry out the search.
  2. If the novel protein is derived from a known allergenic source or it has significant sequence homology with a known allergen, the reactivity of the novel protein with immunoglobulin-E (IgE) from the blood serum of appropriate allergic individuals should be evaluated.
  3. The digestive stability of newly expressed protein(s).
  4. The heat stability of newly expressed protein(s).

7.0 Laboratory animal/livestock feeding trials

Laboratory animal and/or livestock feeding trials may be necessary to address safety and efficacy assessment requirements for plant-derived novel feeds. These studies should be conducted using materials representative of the material that will be used in Canada. In some cases, conducting whole plant feeding trials for the safety assessment may be impractical due to variations in composition and difficulty in feeding sufficient quantities of the substance in question. As such, every effort should be made to fully characterize the plant, plant parts, or in vitro produced proteins with respect to appropriate counterparts and equivalent comparators.

Laboratory animal and/or livestock feeding trials in support of the safety of the plant derived novel feed may be required to address any toxicological concerns arising from an assessment of data submitted under Sections 2.1 to 6.0 of these guidelines, including acute toxicity data.

Feeding trials may also be needed to address issues related to efficacy of the plant derived novel feed such as evidence of the nutritional adequacy and nutrient bioavailability. The necessity of these studies would be determined based on an assessment of data submitted under Sections 2.1 to 6.0 of these guidelines.

In the case of intentional modification of the nutritional composition or functionality, in vitro and/or in vivo studies to support the safety and efficacy of the feed are required.

Authorization of the release of a novel feed for research purposes must be obtained from the Animal Feed Division prior to conducting livestock feeding trials with novel feeds in Canada. For guidance on applying for authorization of the release for research purposes, please refer to Chapter 5 of this document (Regulatory Guidance: Feed Registration Procedures and Labelling Standards).

8.0 Evaluation of environmental safety

An evaluation of the potential impact of the novel feed on the environment is required if any component of the novel feed is released, including excreta from the livestock consuming the feed.

This evaluation may also consider the following criteria, if not reviewed by the CFIA as part of a PNT assessment:

  1. Potential of the plant to become a weed of agriculture or be invasive of natural habitats.
  2. Potential for gene flow from the plant to wild relatives whose hybrid offspring may become more weedy or more invasive.
  3. Potential of the plant to become a plant pest.
  4. Potential impact of the plant or its gene products on non-target species, including humans.
  5. Potential impact of the plant on biodiversity.

9.0 Method of detection and identification requirements

In certain situations where it is required to verify compliance with the conditions of authorization, the CFIA may request the following information be submitted:

If the CFIA identifies that a method of detection and identification is required as part of the Novel feed submission, the CFIA will provide guidance on the information to be provided.

Certain information regarding the detection and identification of a novel feed may be protected under the provisions of the Access to Information Act. All requests for such information are not only subject to the Access to Information Act but to the Privacy Act as well.

10.0 Regulatory decision

Based on the information received, and after evaluating the potential impact on livestock, humans and the environment, the Animal Feed Division will:

  1. Authorize the release of the novel feed where there is minimal risk (note that conditions may be imposed on authorizations to manage risk);
  2. Refuse to authorize the release of the novel feed where its use poses an unacceptable risk, including when there is insufficient data to demonstrate safety and/or efficacy; or
  3. Follow up with the applicant for any additional information that the Animal Feed Division requires to complete the assessment ("deficiency letter"), followed by one of the decisions above. The Animal Feed Division will consider the application closed if no response to the deficiency letter is received within one year from the date of the deficiency letter.

Once a novel feed (including those with a novel trait) has received authorization (i.e., it gets listed in Schedule IV or V or is determined to be substantially equivalent to an ingredient already listed in the Schedules), it is no longer considered to be novel.

11.0 Posting of decision

After a regulatory decision has been made to authorize a novel feed that is a new feed ingredient, a definition for the ingredient will be added to Schedule IV or V of the Feeds Regulations.

After a regulatory decision has been made concerning a feed with a novel trait that is not a new feed ingredient, the Animal Feed Division posts a decision document, which is a record of the assessment, on the CFIA website. The decision document explains the regulatory decision reached by the Animal Feed Division, and the Plant Biosafety Office (PBO) in cases where the novel feed is from a plant source that is also a PNT, following their evaluations.

12.0 New information requirements

If at any time the proponent becomes aware of any new information regarding risk to the environment, livestock or human health that could result from the release of the novel feed the proponent must immediately provide such information to the CFIA.

On the basis of new information, regardless of source, the CFIA will re-evaluate the potential impact on and risk to the environment, including the potential impact on and risk to human or animal health with respect to the livestock feed use. The CFIA may impose additional conditions on the release or use as livestock feed, may change the conditions on these authorizations, or cancel the authorizations and require the proponent to stop the release or use as livestock feed and take any appropriate action necessary to eliminate or minimize the risk.

13.0 Definitions

Biotechnology:
The application of science and engineering in the direct or indirect use of living organisms or parts or products of living organisms in their natural or modified forms (Canadian Environmental Protection Act).
Carrier DNA:
DNA used to expedite the preparation or the transformation of genetic material into a plant but which is itself not part of the construct.
Coding region:
A DNA sequence which can be translated to produce a protein.
Construct:
An engineered DNA fragment (e.g., plasmid) which contains, but is not limited to, the DNA sequences to be integrated into a target plant's genome.
Counterpart:
A feed or feeds selected by the applicant, to which the novel feed will be compared. The chosen counterpart should be, if possible, the closest genetic equivalent to the novel feed and may be a previously assessed and authorized novel feed. Since there will be a range of characteristics among varieties within a species, comparison with several counterparts may show that the novel feed has characteristics within the normal range exhibited by that species. If the closest genetic equivalent to the novel feed is not similar to the novel feed (e.g., in the case of a plant intentionally modified for composition), consideration may also be given to comparison with other products the novel feed was designed to emulate.
Database Citations:
Publicly available sources of nucleotide or protein sequence information. Five commonly used databases and their website addresses are found in the references section.
Environment:
The components of the earth (including air, land and water, all layers of the atmosphere, all organic and inorganic matter and living organisms) and all interacting natural systems that include components referred to above (Feeds Regulations). This includes natural and managed ecosystems which include agricultural ecosystems.
Familiarity:
The knowledge of the characteristics of a plant species and experience with that plant species in Canada.
Feed:
Any substance or mixture of substances containing amino acids, anti-oxidants, carbohydrates, condiments, enzymes, fats, minerals, non-protein nitrogen products, proteins or vitamins, or pelletizing, colouring, foaming or flavouring agents and any other substance manufactured, sold or represented for use:
  1. for consumption by livestock,
  2. for providing the nutritional requirements of livestock, or
  3. for the purpose of preventing or correcting nutritional disorders of livestock, or any such substance for use in any such substance or mixture of substances (Feeds Act).
Gene flow:
The transfer of genetic material by interbreeding from one population of a species to another population (same or related species), thereby changing the composition of the gene pool of the receiving population.
Genetic Modification:
Any intentional change to the heritable traits of an organism. This includes, but is not limited to: traditional breeding, recombinant nucleic acid techniques, somaclonal variation, electroporation, mutagenesis, and similar techniques.
Insert:
That part of the construct which is integrated into the recipient plant's genome.
Irritant:
Any agent capable of eliciting an abnormally excited or sensitive condition in a body part of a human or other animal.
Livestock:
Means horses, cattle, sheep, goats, swine, foxes, mink, fish, rabbits and poultry and includes such other creatures as may be designated by regulation as livestock for the purposes of the Feeds Act.
Noncoding region:
DNA sequences which lie outside of an open reading frame and which are not translated to become part of a protein. These might include scaffold attachment regions, promoters, leader sequences, enhancers, introns, terminators, and any other sequences that are used for gene expression either in the plant or other hosts, such as origins of replication, transposon elements, T-DNA borders, lox sequences, etc.
Novel Feed:
Means a feed comprising an organism or organisms, or parts or products thereof, that:
  1. is not set out in Schedule IV or V, or
  2. has a novel trait (Feeds Regulations).
Novel Trait:
Means a characteristic of the feed that:
  1. has been intentionally selected, created or introduced into the feed through a specific genetic change, and
  2. based on valid scientific rationale, is not substantially equivalent in terms of its specific use and safety both for the environment and for human and animal health, to any characteristic of a similar feed that is set out in Schedule IV or V (Feeds Regulations).
Plant with Novel Traits (PNT):
A plant containing a trait not present in plants of the same species already existing as stable, cultivated populations in Canada, or containing a trait present at a level significantly outside the range of that trait in stable, cultivated populations of that plant species in Canada.
Safety Assessment:
Encompasses hazard identification, risk estimation, and risk evaluation and management.
Stability:
The ability of the trait to be expressed in the modified plant line and plant lines derived there from in a consistent, reliable, and predictable manner.
Substantial Equivalence:
Equivalence of the feed from a plant, within a particular plant species, in terms of its specific use and safety to the environment and human and animal health, to those in that same species, that are in use and generally considered as safe in Canada, based on valid scientific rationale.
Trait:
The phenotypic characteristic(s) conferred to the recipient plant by specific genetic changes.
Transgenic Plant:
A plant in which one or more genes, genetic constructs, or traits have been introduced using recombinant DNA techniques, which could be considered to include the insertion of genetic material from the same or different species.
Unintended Effects:
In addition to intended changes caused by the insertion/mutation of defined DNA sequences during preparation of a genetically modified plant, additional unintended traits may be acquired or existing traits could be lost or modified. These unintended effects may be deleterious, beneficial or neutral with respect to health of plant and safety to livestock and humans.
Vector:
An autonomously replicating DNA molecule into which foreign DNA is inserted and then propagated in a host cell.

14.0 References

GenBank: An annotated collection of all publicly available DNA sequences maintained by the National Institute of Health (NIH).

DNA Data Bank of Japan: The officially certified DNA bank of Japan, which collects DNA sequences from researchers.

EMBL Nucleotide Sequence: A database of DNA and RNA sequences collected from the scientific literature, patent applications, and directly submitted from researchers and sequencing groups.

FAO: The Food and Agriculture Organization of the United Nations leads international efforts to defeat hunger. The FAO acts as a neutral forum where nations meet as equals to negotiate agreements and debate policy. The FAO is also a source of knowledge and information.

FARRP Allergen Database: A database containing a list of unique known and putative allergens that were identified by searching publicly available protein databases.

OECD: The Organisation for Economic Co-operation and Development is an international organisation that accepts the principles of representative democracy and free-market economy. The OECD publishes reports that contain technical information for use in the regulatory assessment of products of biotechnology. These documents are mutually recognized among member countries of the OECD.

SWISS-PROT Protein Sequence Data Bank: A database of protein sequences produced collaboratively by Amos Bairoch (University of Geneva) and the EBI.

Appendix I: Molecular characterization requirements for feeds with novel traits derived from plants developed using recombinant DNA techniques

For plants developed using recombinant DNA techniques, please provide the following:

  1. For each event derived through transgenic methods, a unique identifier for each line must be designated according to the "OECD Guidance for the Designation of a Unique Identifier for Transgenic Plants" - PDF (40 kb).
  2. A description of the genetic element responsible for the novel trait.
  3. State the objective of the modification, e.g., novel herbicide tolerance, male sterility/restoration, etc.
  4. A description of, and references for, the method used to introduce the novel trait(s).
    1. Information on the nature and source of the DNA used to modify the plant (e.g., carrier DNA, vectors, helper plasmids).
    2. For Agrobacterium-mediated transformation, the strain designation of the Agrobacterium used during the transformation process, and the method by which the Ti plasmid based vector was disarmed, and whether Agrobacterium was cleared from the transformed tissue.
    3. For transformation systems other than Agrobacterium:
      • Whether the system utilizes a pathogenic organism or nucleic acid sequences from a pathogen, and, if so;
      • how any pathogenesis-related sequences were removed prior to transformation.
  5. A description of the genetic material potentially delivered to the host plant.
    1. A summary of all genetic elements which comprise the vector including coding regions, and non-coding sequences of known function. For each genetic element a citation where these functional sequences were described, isolated, and characterized (publicly available database citations are acceptable), indicate:
      • The size and identity of the genetic element;
      • The location and orientation in the vector;
      • The function in the plant;
      • The source (scientific and common name of the donor organism);
      • Whether the genetic element is responsible for disease or injury to plants or other organisms, or if its products are known toxicants, allergens, pathogenicity factors, or irritants.
    2. If there has been a significant modification that affects the amino acid sequence of the protein to be expressed in the plant, the specific amino acid changes should be identified. Indicate whether the modifications are known or expected to result in changes to post-translational modification or affect sites critical to the structure or function of the gene product.
    3. A map of the vector with the location of sequences described above that is sufficient to be used in the analysis of data supporting the characterization of the DNA, including, as appropriate, the location of restriction sites and regions used as probes and/or primers used for polymerase chain reaction (PCR).
  6. A characterization of the DNA inserted in the plant, including:
    1. Number of insertion sites;
    2. Data that demonstrate if complete or partial copies are inserted into the plant's genome
    3. For noncoding regions associated with the expression of coding regions:
      • Whether or not plant promoters are inserted intact with the coding regions whose expression they are designed to regulate;
      • DNA analysis may be necessary for introns, leader sequences, terminators, and enhancers of plant-expressible cassettes;
      • DNA analysis may be necessary for promoters and other regulatory regions associated with bacteria-expressible cassettes
    4. For noncoding regions which have no known plant function and are not associated with expression of coding regions:
      • DNA analysis may be required for some sequences of known function (e.g., ori V and ori-322, T-DNA borders of Agrobacterium, and bacterial transposable elements)
      • DNA analysis is not required for any remaining sequences of the plasmid backbone when the plasmid is well characterized
    5. Where appropriate, sequence data of the inserted material and of the surrounding regions (sequencing information may be informative in some cases, i.e., to fully characterize a partial or rearranged DNA insert).

      Protein or RNA characterization may not be required for fragments of genetic constructs not expected to be functional in the plant (e.g., fragments of selectable marker genes driven by bacterial promoters).

    6. Consider if a fusion protein could be produced if a fragment of a coding region designed to be expressed in the plant is detected, and in which tissues it may be located.

Appendix II: Novelty determination guidance

It is the responsibility of the proponent to consider whether they have developed a novel feed or not. When proponents are determining whether a plant-derived feed is novel or not, there are some basic questions they can consider.

Q1. Is the plant-derived feed listed in Schedule IV or V of the Feeds Regulations?

No, novel feed.

For feeds with novel traits:

Q2. Is the feed derived from a plant that has one or more characteristics that are new or outside of the conventional range for that plant species?

Yes, novel feed

Proponents should use their expertise and relevant scientific literature to determine the range of a selected trait based on Canadian experience to determine if they have created a feed with a novel trait. A substantive change in agronomic, nutritional or compositional characteristic(s) of a plant that are new or outside the accepted range for a given species would constitute a novel trait, thus triggering the requirement to apply for authorization of the release of feed derived from the plant. For example, feed derived from a new wheat line with an increase in yield that is similar to increases seen historically in wheat lines cultivated in Canada would not be considered novel. Feed derived from a new insect-resistant canola line with resistance many fold higher than that displayed by plants currently grown in Canada would be a feed with a novel trait. Feed derived from a new soybean line with a substantive increase in methionine would be a novel trait.

Q3. Is the feed derived from a plant that is a descendant, produced through intraspecific crossing, of a plant previously authorized for livestock feed use?

Yes, not novel

After a plant is authorized for use as livestock feed, all progeny of the modified plant and sister lines, which have been derived from the original modified plant, are also authorized for livestock feed use provided that the proponent has determined that:

The CFIA may ask the proponent to provide evidence supporting these conclusions.

Q4. Is the feed derived from a plant that is a descendant, produced through interspecific crossing, of a plant previously authorized for livestock feed use?

Yes, likely novel

For feeds with novel traits, it is the presence of the novel trait that triggers regulatory oversight under the Feeds Regulations, not the method used to introduce the trait. As such, the transfer of a previously assessed and authorized trait into a recipient plant species through interspecific crossing can result in the creation of a novel feed. This means that the recipient species could require assessment and authorization prior to use as a livestock feed.

Subsequent to authorization by the Animal Feed Division for livestock feed use, the proponent may not be required to apply to the Animal Feed Division for authorization of additional lines derived through similar means (i.e. interspecific crossing), provided that the proponent has determined that:

The CFIA may ask the proponent to provide evidence supporting these conclusions.

Q5. Is the feed derived from a plant possessing previously authorized traits that have been intentionally stacked?

Yes, notification of the Animal Feed Division required

The Animal Feed Division is to be notified at least 60 days prior to the intended livestock feed use of plants having stacked traits resulting from intentional intraspecific or inter-specific crosses between novel plants already authorized for livestock feed use. Following the proponent's notification, the Animal Feed Division will issue a letter (within 60 days of notification) informing the proponent of any concerns the Animal Feed Division may have regarding the livestock feed use of the plant with stacked traits. The Animal Feed Division reserves the right to request and review data to support the safe use of the modified plant in livestock feed. Stacking of traits with potential incompatible management requirements, possible negative synergistic effects, or where feed use of the plant is altered may result in the requirement for an assessment. Until all livestock feed concerns have been resolved, the modified plant must not be used as livestock feed.

Q6. Is the feed derived from a plant that is a re-transformation or re-mutation of a plant previously authorized for use as a livestock feed?

Yes, may be novel

Re-transformation is considered to be the transformation of a plant with the identical construct(s) as a previously authorized plant of the same species and confers the same trait. Re-mutation is considered to be the mutation of the same gene in a plant as a previously authorized plant of the same species which confers the same trait. Such plants may not require an application to the Animal Feed Division, provided that:

Note that if a proponent determines that a feed is not novel the information to support the determination must be retained and presented to the CFIA on request.

Consultation with the Animal Feed Division is highly recommended for any proponents experiencing difficulty in determining novelty.

2.7 Guidelines for the assessment of novel feeds: microbial sources

1 Introduction

1.1 Scope

These guidelines (Section 2.7 of RG-1, Regulatory Guidance: Feed Registration Procedures and Labelling Standards) apply to novel feeds composed of microorganisms, or products derived from them, which have not previously been authorized by the Canadian Food Inspection Agency (CFIA) for use as livestock feed in Canada and/or have a novel trait.

Prior to the release of a novel feed (i.e., the manufacture, sale or import or use as livestock feed), authorization from the Animal Feed Division (AFD) of the CFIA is required.

This section of the RG-1 provides guidance for the preparation of an application for the authorization of the release of a novel microbial feed. It includes criteria that will be considered in the assessment of the safety and efficacy of a novel feed. It is not intended to explicitly define all of the data that might be required in the course of the assessment.

Learn more about novel feeds, application procedures and AFD activities

Please be aware that the information provided in these guidelines is not exhaustive and will be updated as appropriate to reflect current scientific knowledge. For further clarification, applicants are strongly encouraged to consult with the AFD. For all purposes of interpreting and applying the law, applicants are invited to consult the official version of the relevant Acts and Regulations.

1.2 Microbial feed products

Microbial products commonly used in livestock feeds are classified into one of two categories:

1.3 Classification of viable microbial products (feeds, drugs, or veterinary biologics)

A viable microbial product or products thereof for direct livestock consumption is classified as a feed, a drug, or a veterinary biologic, based on the product label claims. The onus is on the company producing the product to determine, and support, the appropriate classification of its product in a satisfactory manner.

When product label claims, company advertising claims, or scientific literature indicate that the viable microbial feed product is a drug or a veterinary biologic, applicants are advised to consult with the appropriate government body (i.e., the Veterinary Drugs Directorate at Health Canada, or the Canadian Centre for Veterinary Biologics at the CFIA). For more information about the classification of viable microbial products, including the types of claims associated with each classification and how to contact the appropriate organization, please consult Section 3.22, Viable Microbials and Yeasts.

More information about the classification of drugs vs. feeds is also available in the Guidance document on classification of veterinary drugs and livestock feeds, available on the Health Canada website.

1.4 Regulation of novel feeds

The AFD evaluates and regulates all feed ingredients, including novel feeds, in the same manner. Any feed ingredient that is new (i.e., is not already listed in Schedules IV or V of the Feeds Regulations), or has been modified such that it differs significantly from a conventional ingredient, is required to undergo a pre-market assessment and approval. The purpose of all feed assessments is the same: to ensure that the feed ingredient is safe (in terms of animal health, human health via residues in food and worker/by-stander exposure, and the environment) and effective for its intended purpose prior to marketing. The evaluation also ensures that the feed is accurately defined in the Feeds Regulations and is labelled appropriately for its safe, effective use and for consumer protection.

1.5 Research with novel microbial feed products

If novel microorganisms, or parts or products derived from them, are not ready for commercialization but will be used in livestock feeding trials or in pilot scale feed development research trials, the AFD must be notified and an application for Authorization for the Release of Novel Feeds for Research Purposes must be submitted. Please refer to RG-1, Chapter 5 – Research with Livestock Feeds for more information.

1.6 Assessment principles

For general information about how applications for registration for feed products are assessed, please refer to Chapter 1 of the RG-1.

The AFD conducts novel livestock feed safety assessments based on familiarity and substantial equivalence. Familiarity and substantial equivalence are two key concepts developed by the OECD (1993) and further elaborated by the FAO/WHO (2000).

Familiarity can be defined as the knowledge of the properties and characteristics of a microbial product and its uses and applications in livestock feeds in Canada, i.e., those listed in Schedules IV or V of the Feed Regulations. Substantial equivalence is based on the comparison of a novel product to an appropriate counterpart, taking into consideration both intended and unintended effects. A Degree of Equivalence can be established for novel microbial products if the intended uses, composition and functional properties of the product are substantially equivalent and the production strains are substantially equivalent in terms of safety to animal health, human health, and the environment. Once familiarity and a degree of equivalence are established, the safety assessment process focuses on the differences between the novel product and its counterpart and their impact on the safety of the novel product.

Essential considerations in the assessment of novel feeds from microbial sources include the safety of the production microorganism and microbial products to humans, animals and the environment. Some areas of consideration during the assessment include:

Where microorganisms may contaminate meat, Health Canada may require an assessment of the safety of the product as a novel food.

1.7 The application process

General administrative information regarding the procedures for application for feed registration can be found in RG-1, Chapter 1, Administrative Requirements for Registration and Approval of Livestock Feed, Specific requirements are found in Part 2 below, and in the attached checklist.

Certain microbial products or ingredients have specific data requirements necessary for supporting safety or efficacy claims. Please refer to the appropriate guidance documents for additional information:

2 Characterization of the microbial product

The submission package must include a summary to provide the reader with an overview of the product, its purpose, its applications and effects on the target species, the identity of the production organism, and, where applicable, the identity of the donor micro-organism and the modification method(s).

All experimental work must be conducted in accordance with sound scientific principles and Good Laboratory Practices. The methods used to generate the experimental data must be submitted, or referenced if published.

All data submitted in support of the safety assessment, including tables, maps, figures, graphs, analytical gels and blots, should be of high quality, comparable to that accepted by reputable peer-reviewed scientific journals. Particular attention should be paid to the sensitivity and specificity of analytical methods. Appropriate controls and reference material used in the analyses and data demonstrating the reproducibility, sensitivity or precision of a method must be submitted. Statistical analyses must be conducted using appropriate techniques and raw data must be made available upon request. Literature reviews submitted in support of the safety of a product must include all references cited in the text. Where data from published literature is cited in support of the safety of the product, a copy of the publication or a journal Web site for the full text article must be provided.

Reviewers' checklists (see Appendix II of this document) provide guidance to applicants in the preparation of quality data. It is recommended that applicants review the appropriate checklist(s) for analytical techniques while assembling a data package in support of a submission.

The manufacturing process must ensure the maintenance of a pure and genetically stable strain, and the production of a consistent product, free from contamination.

The toxicological and physicochemical properties of the microbial product must be well-characterized, since the microbial preparation may range from highly purified compounds to crude fermentation product, containing media residues, cellular components and various products and by-products of microbial metabolism.

Oral toxicity studies and genotoxicity tests may be required, to confirm the absence of harmful substances that may be produced as co-products during the fermentation process or may be present as contaminants.

When the feed microbial product consists of, or is derived from, a genetically modified microorganism, additional considerations would include: the safety of the donor organism (where applicable); the characterization of the genetic modification and the novel trait; the properties of the modified organism; any unintended effects and/or secondary effects; and the disruption or alteration of metabolic pathways induced by the genetic modification. These alterations may be beneficial to the feed, may have no effects, or may introduce toxic or allergenic com­pounds and changes in safety characteris­tics of the feed. For a live microorganism, the properties conferred as a result of the genetic modification must be evaluated, and the effect of these properties on the survival, competitiveness and survival fitness of the microorganism in the GI tract and in the environment must be determined.

2.1 Product description and use

The following lists the criteria that are considered in the assessment process, and is intended to provide guidance for the preparation of an application for authorization. Please submit all applicable information:

  1. The identity of the product and a description, including:
    • proposed name(s)
    • type of product and identity of the active ingredient
    • product formula, if consisting of a mixture, listing the ingredients and their proportion by weight
    • physical state of the product, including any processes such as granulation or encapsulation
    • purpose of the active ingredient/product
    • mode of action of the active ingredient
    • taxonomic identity of the production microorganism, including strain number
  2. Unit amount/package size
  3. Product label
  4. Directions for use, including:
    • mixing information, if used with other products
    • suggested rates, timing, intervals of feeding
    • identification of species of intended use
    • distribution, storage and handling information
    • strategies for reuse, resale and disposal of unused product
  5. Notifications and/or regulatory approvals of other government agencies and international agencies
  6. A list of all ingredients, including those used in manufacturing, and a description of their mode of action or purpose
  7. Material Safety Data Sheets (MSDS) for the product and the ingredients
  8. A detailed description of the manufacturing process, including:
    • a flow chart of the manufacturing process, from strain revival to product packaging, describing the steps in the preparation of the microbial product
    • the composition of the growth media, from the initial inoculation steps to the production level bio-reactor
    • the fermentation conditions
    • a detailed description of the separation and purification steps, including all processing aids that are used during the manufacturing process
    • the batch-to-batch variation in the concentration of the active ingredient and the impurities
    • quality control/quality assurance procedures and protocols for purity of the production microorganism prior to and during manufacturing, production parameters, contamination, consistency and conformity of the final product, and the methods used to monitor genetic drift of the production organism
    • methods for treatment and/or disposal of by-products and waste products resulting from the fermentation process. Data supporting the efficacy of the treatment and/or disposal method should be provided. This requirement is specific to microbial fermentations conducted in Canada
    • a statement confirming the application of Canadian Biosafety Guidelines for lab use, and National Institutes of Health (NIH)/Good Large Scale Practices (GLSP), for large scale manufacturing, where applicable. A description should be provided on how the facility meets every point described in CBSG or NIH to ensure adequate containment
  9. The composition of the final product, including:
    • the proportion and concentration of ingredients used to formulate the final product, including the amount of source material derived from the fermentation process, (example, 90% rice hulls (carrier), 10% fermentate)
    • the percentage purity of the source material derived from the fermentation process, example, fermentate is composed of 70% active ingredient and 30% media residue. The purity of other ingredients in the product must also be provided
    • the amount of active ingredient in the final product in ppm (parts per million) or per cent, or in colony forming units (CFU)/g or viable cells/g (depending on the enumeration method) when the active ingredient is a live organism. When the product is composed of a mixture of microorganisms, each strain must be described separately, and the numbers of each strain in the product must be provided
    • identification of any extraneous material likely to occur in the final product
  10. Certificates of Analysis for three production batches or pilot scale batches for the active ingredient, demonstrating identity and concentration. Appropriate controls or standards must be included, if applicable. The methods of analysis must be described, or referenced if published. As an example, for an amino acid, the data should demonstrate the concentration and identity of the amino acid as compared to a mixture of amino acid standards
  11. Data supporting the presence or absence, or viability or non-viability, of the production strain in the final product, depending on the nature of the product and the specifications
  12. Data supporting a shelf life of at least 12 months from the date of manufacture, as shown for a minimum of three production batches of the product
  13. A list of all heavy metal, chemical and biological contaminants, either inherently present or introduced via the process, including:
    • certificates of Analysis for biological contaminants in three production batches of the product for total plate counts, pseudomonads, Salmonella, Staphylococcus, E. coli, coliforms, yeasts and moulds (with no Aspergillus flavus or Fusarium spp. detected)
    • certificates of Analysis for heavy metals, chemicals and for mycotoxins where applicable
    • analytical methodologies used to quantify contaminants

2.2 Characterization of the production microorganism

The following information must be provided:

  1. Taxonomic classification based on a recognized nomenclature organization or refereed scientific publications, to allow for the complete identification of the microorganism, including the following: genus, species, subspecies, strain and/or type, as well as any substantiated changes in classification, and internal identification numbers if applicable. Information on the methods used to substantiate the identification along with all test data should be provided (details explained in paragraph D below). A Certificate of Analysis should be included, confirming the identification of the microorganism from three recent production batches
  2. Common name(s), including abbreviations or acronyms, as well as names used in other countries
  3. Accession numbers or other information from a recognized culture repository, if available, and a copy of the deposit certificate for the organism (and/or for the parental strain if the production organism has been genetically modified)
  4. Criteria, methodology and analytical data to substantiate identification and taxonomic assignation of the microorganism, including phenotypic traits (morphology, substrate usage, fermentation products, etc.) and phylogenetic analysis (16S or 23S ribosomal RNA sequencing, or other molecular taxonomic methods). Immunological tests (example, radioimmunoassay [RIA]) may be necessary depending on the source of the microorganism. For pure cultures, the information submitted should be sufficient to distinguish the microorganism from related microorganisms, and to describe its relationship to pathogens. For mixed cultures, it is necessary to provide the identification and characterization information for each strain present in the mixture along with the methods and criteria used to select and enumerate each strain. In exceptional circumstances, complex mixtures may be considered on a case-by-case basis
  5. The strain's origin and following history from point of isolation to acquisition (example: original habitat, environmental isolate, certificate of origin, clinical isolate, culture collection) and development (example: selection procedures, culture maintenance)
  6. Reports and documentation on any history of use or hazards of the strain in agriculture, food production and other industrial practices
  7. Other documented uses of the strain and closely related strains or species and information on exposure routes
  8. Any reports of disease, opportunistic pathogenicity, virulence factors, and/or toxin production associated with the strain or closely related species. Where related strains or species of microorganisms are capable of producing adverse effects, the absence of such effects should be demonstrated in the production organism; this may include data to demonstrate that genes responsible for the adverse effects in related species have been identified and their absence or suppression in the production strain has been shown
  9. Detailed description of any inherent antimicrobial activity
  10. Detailed description of any inherent antimicrobial resistance factors present in the strain and, when necessary, the characterization of the genetic determinants of the antimicrobial resistance, example, whether they are associated with mobile genetic elements or integrated in a stable manner into the chromosome. Scientific rationale, with supporting evidence, is to be provided on the safety concerns, or lack thereof, of resistance elements present in the strain

    If any antimicrobial resistance is detected, the potential impact on human and animal health must be described. A review of the scientific literature should be conducted to document any observed resistance in other strains of the same species for target antibiotics

    A national standard should be followed (example,European Food Safety Authority) when establishing minimum inhibitory concentration (MIC) values for the notified strain. Control strains should be included with average MIC values for the species and pathogenic members of the genus

  11. Information on the presence of mobile genetic elements (example, insertion sequences, transposons, plasmids, prophages)
  12. If the organism has not been genetically modified, a statement to that effect must be submitted

2.3 Development of the modified microorganism

This subsection is for consideration only if the organism has been genetically modified by recombinant nucleic acid techniques, deletion, rearrangement or suppression of native DNA, classical mutagenesis techniques, selective pressure, or other techniques.

Sufficient information on the process used to effect the genetic modification should be submitted to enable the characterization of the modified microorganism, and permit comparison with the conventional or unmodified counterpart. The identification of all genetic material introduced, modified or deleted in the recipient organism and scientific analysis of the data supporting the molecular characterization should allow an assessment of both safety and potential secondary and unintended effects. These effects could be caused by insertional activation or inactivation of genes, transcription of vector sequences, and/or the impact of foreign gene products on the metabolism of the host, leading to altered levels of existing gene products or metabolites.

Newly expressed material, resulting from introduced genetic material or mod­ified native material, should be fully characterized, and similarity to products from traditional sources should be described where appropriate. If novel constituents (other than those resulting from the intentional modification of genetic material) are identified, further studies are required to characterize the unintended effects and their impact on the safety of the production organism and the microbial product. It should be demonstrated that introduced changes do not cause adverse effects, changes are stable, and potential gene transfer to other species is not enhanced. Where genetic modifications alter the expression of a traditional metabolite or constituent, sufficient information on the anabolic or catabolic pathways should be provided to enable an assessment of possible secondary effects on related pathways and metabolite production.

The following information must be provided:

  1. Description of the Donor Organism(s) and Intermediate Organism(s) (where applicable), including:
    • taxonomic names, based on the most current recognized nomenclature organization or refereed scientific publications, to allow for the complete identification of the donor organism, including any substantiated changes in nomenclature and any previous nomenclatures
    • common name(s), including alternatives, abbreviations or acronyms, as well as names used in other countries
    • accession numbers or other information from a recognized culture repository, if applicable, and the deposit certificate
    • criteria and methodology to substantiate identification and classification
    • origin of the donor organism(s) and intermediate organism(s), including history during development (example, selection procedures, culture maintenance)
    • reports and documentation on any past and present uses of the donor organism, or closely related organisms in feeds, foods or other applications, and information on exposure routes, hazards, etc
    • any reports of disease, pathogenicity, including opportunistic pathogenicity, virulence factors or toxin production by the donor organism or by closely related organisms
  2. Description and Characterization of the Genetic Modification, including:
    • description of the genetic modification, including a list of the genes introduced, modified or deleted, their function and the resultant novel traits
    • purpose of the modification and the intended properties conferred to or lost by the genetically modified microorganism as a result of the modification
    • detailed description of the recipient strain construction/modification process, including:
      • a) a flow diagram, including the names of the donor, recipient and vector DNA and the steps (linked in the order of their development) used to construct the final modified microorganism
      • b) a description of the method(s) used to introduce, delete or modify the genetic material and the resultant novel trait(s), including references or citations
      • c) when vectors are used during the genetic modification, the following must be provided:
        • the name and source of the vector(s)
        • the origin and characterization of the sequences of the vector(s), including regulatory elements (example, promoters, modifiers, enhancers, signal peptides and terminators), replicons, selective marker genes, linkers and any foreign DNA, in case the vector or DNA sequences from the vector are part of the genetically modified microorganism. Citations or references should be included on the development of the vector
        • the natural pathogenicity and infectivity of the vector
        • the method used to disarm the vector, such as removal of antimicrobial resistance markers or inactivation of transposons and other mobile elements, if applicable
      • d) a description of the intermediate recipient organisms used to produce or process DNA prior to introduction into the final recipient organism; and
      • e) a physical map of each genetic construct, detailing size and positions of all coding and non-coding sequences, all regulatory elements, origins of replication and transfer, marker genes, location of restriction sites, primers used for PCR analysis, and regions used as probes. A table identifying and referencing each component should be included with the map. Appendix III is provided as a model to follow for the submission of vector characterization data

2.4 Characterization of the novel trait(s)

Provide the following information about the new modification, whether introduced or modified native material:

  1. a description of the morphological and physiological characteristics, growth patterns and growth rates of the modified microorganism as compared to the unmodified parental (or isogenic) strain
  2. data that demonstrate the genetic stability of the modified organism and that all novel traits are expressed and inherited in a stable manner
  3. the gene product(s), breakdown products, by-products and their metabolic pathways, and the analysis of transcripts or expression products to identify any new substance that may be produced
  4. the function of the gene product(s)
  5. the phenotypic description of the new trait
  6. the level and the site of expression of the gene product(s) including those of the marker gene(s), if applicable
  7. a determination whether gene expression is constitutive or inducible, with a description of the inducing agents
  8. the activity of the gene product(s), breakdown products and by-products in the host microorganism, and any resulting changes to existing metabolic pathways (including altered accumulation and storage patterns)
  9. for microorganisms used for the processing or treatment of feeds (example, forage inoculants, acidifiers), the activity of the gene product(s), breakdown product(s), and by-product(s) should be examined under the conditions of use
  10. for all coding regions inserted or modified, expressed protein characterization data, including:
    • any modifications to the amino acid sequence and properties of the protein, or post-translational modifications to the expressed protein, such as glycosylation or phosphorylation, which may have occurred when the coding sequence has been modified or transferred from one organism to another
    • where protein structure has been modified, data to demonstrate how the amino acid modifications affect sites critical to structure or function, and amino acid sequence homology of the novel protein to the amino acid sequences of closely related proteins from the same family and to native proteins, indicating the locations of the amino acid differences and whether they occur in variable or conserved regions
    • The following data may be required for novel enzymes:
      • a) chemical name, enzyme commission classification number, and Chemical Abstract Service (CAS) registry number
      • b) a description of the catalytic functions and mode of action
      • c) the substrate specificity for each catalytic function
      • d) known co-factors required for enzyme activity
      • e) pH optimum and profile
      • f) temperature optimum and profile
      • g) kinetic properties for each substrate assayed, including Vmax, Km, and Kcat. Where the catalytic properties of a novel enzyme differ from the properties of the native enzyme, a valid scientific interpretation must be provided, explaining the significance of this difference in activity, based on the variation in the range of activity exhibited by enzymes belonging to the same family
      • h) for structurally modified enzymes, the impact of the modification on the activity of the novel enzyme, as compared to the native enzyme, at a range of pH, temperature and other characteristics that may affect the activity of the enzyme under the conditions of usage
      • i) the stability and metabolic fate of the novel enzyme

3 Inactivation of antimicrobials in feed

Antimicrobials are routinely mixed into livestock feeds. A determination of the potential for their inactivation during storage in a mixed feed, as a result of inherent, acquired or incorporated antimicrobial resistance, should be conducted. Antimicrobial efficacy, in the presence of the microbial product, should be monitored over a reasonable length of time. If biochemical parameters such as pH or moisture content of the feed preclude the expression or activity of the antimicrobial resistance protein, this study may be waived.

4 Animal and human safety

Safety information must be provided to identify the type and severity of potential health risks that the microorganism(s) or product(s) may pose to humans or animals. Where no reported adverse effects are found, describe any steps taken to ascertain the lack of reports of adverse effects (example, the databases searched, the period search, the search strategy and keywords used). Required information may include but is not limited to, the following:

5 Environmental safety

This part of the guidelines describes the requirements concerning biological and ecological characterization of the novel feed. The information should provide sufficient data to evaluate the potential environmental effects, including the environmental fate of the microorganism(s), its metabolites, breakdown product(s), and any by-product(s). The goal is to identify and assess the extent of any potential hazards, i.e., the short and long-term presence of the novel feed in the environment. If factors that control the survival and multiplication of the microorganism are known, it is easier to predict that organism's survival and multiplication under its condition of use. Where no reported adverse effects are found, please provide reasons for potential false-negatives or a lack of information, including a lack of published research, limitations to the databases searched, limitations to the search parameters including timeframes, the search strategy, or the keywords used. Results must be interpreted using a valid scientific rationale. Information should include but is not limited to:

6 Regulatory decision

Based on the information received, and after evaluating the potential impact on livestock, humans and the environment, the AFD will:

Once a novel feed (including those with a novel trait) has received authorization (i.e., it gets listed in Schedule IV or V or is determined to be substantially equivalent to an ingredient already listed in the Schedules), it is no longer considered to be novel

7 New information requirements

If at any time the proponent becomes aware of any new information regarding risk to the environment, to livestock, or to human health that could result from the release of the novel feed, they must immediately provide this information to the CFIA. On the basis of any new information, regardless of the source, the CFIA will re-evaluate the potential impact on and risk to the environment, including the potential impact on and risk to human or animal health, with respect to the livestock feed use. The CFIA may impose additional conditions respecting the release of the novel feed or its use as livestock feed, change the conditions respecting these authorizations, or cancel the authorizations and require the proponent to stop the release of the novel feed or its use as livestock feed and take any appropriate action necessary to eliminate or minimize the risk.

These Guidelines for the Assessment of Novel Feeds: Microbial Sources, have been prepared to provide guidance regarding the submission of an application for the authorization of the release of a novel feed, as may be required under the Feeds Regulations. Please be aware that the information provided in these guidelines is not exhaustive and will be updated as appropriate to reflect current scientific knowledge. For further clarification, applicants are strongly recommended to consult with the CFIA. For all purposes of interpreting and applying the law, applicants are invited to consult the official versions of the relevant Acts and Regulations.

8 Definitions

Biotechnology:
The application of science and engineering in the direct or indirect use of living organisms or parts or products of living organisms in their natural or modified forms (Canadian Environmental Protection Act).
Carrier DNA:
DNA used to expedite the preparation or the introduction of genetic material into a microorganism but which is itself not part of the construct.
Codex Alimentarius:
(Latin for Food Law): International organization created in 1963 by the UN's Food and Agriculture Organization (FAO) and World Health Organization (WHO) to develop food standards, guidelines and related texts, such as codes of practice, under the Joint FAO/WHO Food Standards Program. The main purposes of this Program are protecting the health of consumers and ensuring fair trade practices in the food trade, and promoting coordination of all food standards work undertaken by international governmental and non‑governmental organizations (Codex alimentarius)
Coding region:
A DNA sequence which acts as the template on which mRNA is synthesized during transcription – the sequence can be translated to produce a protein. This region is composed of the coding strand (the sense strand, or the plus (+) strand) whose sequence is complementary to that of mRNA, and the non-coding strand (complementary to the coding strand) which is also called the antisense strand or the minus (-) strand.
Construct:
An engineered DNA fragment (example, a plasmid) which contains, but is not limited to, the DNA sequences to be integrated into a target microorganism's genome.
Counterpart:
A microbial product selected by the applicant to which the novel product will be compared. The chosen counterpart should be, if possible, the closest genetic equivalent and may be a previously assessed and authorized novel feed. If the closest genetic equivalent is not similar to the novel feed, consideration may also be given to comparison with those products the novel feed was designed to emulate.
Database citations:
Publically accessible sources of nucleotide or protein sequence information. Five commonly used databases and their website addresses are:
GenBank:
An annotated collection of all publicly available DNA sequences maintained by the National Institute of Health (NIH).
DNA Data Bank of Japan:
The officially certified DNA bank of Japan, which collects DNA sequences from researchers.
EMBL Nucleotide Sequence:
A database of DNA and RNA sequences collected from the scientific literature, patent applications, and directly submitted from researchers and sequencing groups.
UniProtKB/Swiss‑Prot Protein Knowledgebase:
A database of protein sequences produced collaboratively by Amos Bairoch (University of Geneva) and the EBI.
FARRP Allergen Protein Database:
A database containing a list of unique known and putative allergens that were identified by searching publicly available protein databases. This database is managed by a panel of scientists and clinicians who are actively involved in reviewing data for inclusion of proteins in the database by comparing peer reviewed publications supporting the classification of the proteins as allergens or putative allergens following predetermined guidelines.
Drug:

Includes any substance or mixture of substances manufactured, sold or represented for use in

  • the diagnosis, treatment, mitigation or prevention of a disease, disorder, abnormal physical state, or its symptoms, in human beings or animals
  • restoring, correcting or modifying organic functions in human beings or animals
  • disinfection in premises in which food is manufactured, prepared or kept (Food and Drugs Act)

A nutritionally-enhanced feed is considered a feed when it is for the purpose of providing nutritional requirements or preventing or correcting nutritional disorders. A nutraceutical or "functional feed" with health benefit claims may be considered a feed or a drug based on the mode of action and the nature of the active ingredient. In this case, the Feed Program assesses the registration application in the preliminary review stage and a decision is made based on the information provided by the applicant.

Environment:

Means the components of the Earth and includes

  • air, land and water
  • all layers of the atmosphere
  • all organic and inorganic matter and living organisms
  • the interacting natural systems that include components referred to in paragraphs a to c. (Feeds Regulations)
Familiarity:
The knowledge of the characteristics of a feed and experience with the use of that feed in Canada.
Feed:

Any substance or mixture of substances containing amino acids, anti-oxidants, carbohydrates, condiments, enzymes, fats, minerals, non-protein nitrogen products, proteins or vitamins, or pelletizing, colouring, foaming or flavouring agents and any other substance manufactured, sold or represented for use

  • for consumption by livestock
  • for providing the nutritional requirements of livestock
  • for the purpose of preventing or correcting nutritional disorders of livestock, or any such substance for use in any such substance or mixture of substances (Feeds Act)
Genetic Modification:
Any intentional change to the heritable traits of an organism. This includes, but is not limited to, change brought about by: recombinant nucleic acid techniques, somaclonal variation, electroporation, artificially induced mutagenesis, and similar techniques.
Genetically Modified Microorganism (GMO):
For the purposes of this document, means bacteria, yeast and filamentous fungi in which the genetic material has been changed through a genetic modification.
Insert:
That part of a construct which is integrated into the recipient microorganism's genome.
Irritant:
Any agent capable of eliciting an abnormally excited or sensitive condition in a body part of a human or other animal.
Livestock:
Means horses, cattle, sheep, goats, swine, foxes, mink, fish, rabbits and poultry and includes such other creatures as may be designated by regulation as livestock for the purposes of [the Feeds Act]. (Feeds Act).
Microorganisms:
Commonly understood to include algae, bacteria, fungi (yeasts and moulds), protozoa and viruses.
Noncoding region:
DNA sequences which lie outside of an open reading frame and which are not translated to become part of a protein. These might include scaffold attachment regions, promoters, leader sequences, enhancers, introns, terminators, and any other sequences that are used for gene expression either in the microbe or other hosts, such as origins of replication, transposable elements, T-DNA borders, lox sequences, etc.
Novel Feed:

Means a feed, comprising an organism or organisms, or parts or products thereof, that:

  • is not set out in Schedule IV or V [of the Feeds Regulations]
  • has a novel trait. (Feeds Regulations)

All feed ingredients approved for use in Canada are listed in Schedules IV or V of the Feeds Regulations. A feed not originating from an organism or products thereof, and not listed in Schedule IV or V of the Feeds Regulations, is considered "Novel" (example, a flocculant chemical ingredient, or tree bark powder ingredient), and must be assessed for both safety and efficacy. However, a feed is considered "New" in cases where it originates from an approved ingredient (example, a purified source of an existing ingredient). In these cases a safety assessment may not be required for ingredient approval. The Feed Program assesses whether a feed is Novel or New based on the information provided by the applicant.

Novel Trait:

In respect of a feed, means a characteristic of the feed that

  • has been intentionally selected, created or introduced into the feed through a specific genetic change
  • based on valid scientific rationale, is not substantially equivalent, in terms of its specific use and safety both for the environment and for human and animal health, to any characteristic of a similar feed that is set out in Schedule IV or V. (Feeds Regulations)
OECD Consensus Documents:
Reports published by the Organisation for Economic Co‑operation and Development (OECD) that contain technical information for use in the regulatory assessment of products of biotechnology. These documents are mutually recognized among member countries of the OECD.
Release:
Means any discharge or emission into the environment of a feed or livestock product produced from the feed, or exposure of a feed or livestock product produced from the feed to the environment. (Feeds Regulations)
Stability:
The ability of an introduced trait to be expressed in the genetically modified microorganism through successive generations in a consistent, reliable, and predictable manner.
Substantial Equivalence:
Equivalence of a novel feed from a microorganism, in terms of its specific use and safety to the environment and human and animal health, to that of a feed from the same species that is in use and generally considered as safe in Canada based on valid scientific rationale.
Trait:
The phenotypic characteristic(s) conferred to the recipient microorganism by a genetic modification.
Unintended Effect:
Unintended characteristic, property or trait acquired, modified or lost by a microorganism as a result of a genetic modification. Unintended effects may be beneficial, neutral or deleterious with respect to the properties of a microorganism and its safety to livestock, humans and the environment.
Vector:
An autonomously replicating DNA molecule into which foreign DNA is inserted and then propagated in a host cell.
Veterinary Biologic:

Means

  • a helminth, protozoa or microorganism
  • a substance or mixture of substances derived from animals, helminths, protozoa or microorganisms
  • 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. (Health of Animals Act)

Veterinary biologics include vaccines, bacterins, bacterin‑toxoids, immunoglobulin products, diagnostics kits, and any veterinary biologic derived through biotechnology.

Appendix I: Description of toxicity/infectivity/pathogenicity tests

The purpose of these tests is to assess the capability of novel microbial feeds to elicit toxicity and to cause disease and tissue injury, by invasion of, and multiplication within, animal or human tissues or the environment.

1. Toxicity tests

The type of toxicity data generally considered is outlined below. Please be advised that appropriate positive and negative controls are required for assays when such controls are applicable to the test. The best way to ensure the tests are conducted appropriately, and using the correct in vitro or in vivo model, is to ensure they are done in accordance with standard toxicity test methods as published by the U.S. Food & Drug Administration, the Organisation for Economic Co-operation and Development (OECD), or ASTM International.

2. Acute bacterial or viral infectivity

This test involves intravenous injection of a range of single doses of the active ingredient into an appropriate animal subject. This study provides information on health hazards likely to arise from a single high dose exposure when the skin is bypassed as a barrier. It is useful as an indication of the inherent infectivity of an organism. It would generally only be used for bacteria of unknown infectivity and viruses

3. Acute fungal or protozoan infectivity

This test involves intraperitoneal (into the peritoneum, i.e., the abdominal cavity) injection of a range of single doses of the active ingredient into an appropriate animal subject. This study provides information on health hazards likely to arise from a single high dose exposure when the skin is bypassed as a barrier. It is useful as an indication of the inherent infectivity of an organism. It would generally only be used for fungi and protozoa. If, based on the physical characteristics of the microorganism, testing by the intravenous route is possible, then the intravenous route should be considered for fungi and protozoa instead of the intraperitoneal route

Appendix II: Molecular data checklists

Checklist for northern blot data
Checklist Included?
The Northern blots have a figure number and title.
Lanes are labelled on the blot.

The figure legend describes each lane of the blot, including a description of the following for the RNA that was loaded:

  • the type of material loaded, example, total purified RNA, poly-A RNA, crude prep, total microbial extract
  • the growth conditions of the cells prior to preparation for loading
  • the quantity of material loaded in each lane
The text or the figure legend describes how RNA was extracted prior to Electrophoresis.
The blot has appropriate positive and negative control lanes (positive control might demonstrate hybridization of the probe with itself; negative control might be the unmodified isogenic microorganism).
The gel system and Northern hybridization protocol are described in the text or in a cited literature reference. Any modifications of the cited protocols are described in the submission text.
The position of molecular size standards on the gel is indicated and they cover an appropriate size range for the fragments that are expected to be detected on the blot.
The probe used for the hybridization is described adequately (in the text or in a figure) to enable interpretation of the results.
The methodology or citation for the quantitative analysis (if done) has been provided, and a sufficient number of replicates or samples have been tested to determine whether there are significant differences between samples or treatments.
Any superfluous bands or background signals are properly explained.
Checklist for southern blot data
Checklist Included?
The Southern blot has a figure number and title.
The lanes are labelled on the blot.

The figure legend describes each lane of the blot, including the following for the DNA that was loaded on the gel:

  • type of DNA loaded (example, entire plasmid, restriction fragment)
  • source of DNA loaded
  • restriction digestions of DNA prior to loading gel
  • quantity of material loaded in each lane
The gel has appropriate positive and negative control lanes (positive control might demonstrate hybridization of the probe with itself; negative control might be the unmodified isogenic parent microorganism).
The gel system and Southern hybridization protocol are described in the text or in a cited literature reference, and any modifications of the cited protocols are described in the petition text.
The positions of the molecular size standards are indicated, and they cover an appropriate size range for the fragments that are expected to be detected on the blot.
If an entire plasmid was used as the probe for the hybridization, it must be described adequately in the text or in a figure to enable interpretation of the results.
If a restriction fragment was used as the probe for the hybridization, it must be described adequately in the text or in a figure to enable one to interpret the results.
Any superfluous bands or background signals are explained.
Checklist for RNA dot blot data
Checklist Included?
The dot blots have a figure number and title.
The lanes are labelled on the blots.

Each figure legend describes each lane of the blot, including a description of the following for the RNA that was loaded:

  • the type of material loaded (example, total purified RNA, crude prep, total microbial extract)
  • the source of the material loaded
  • the quantity of material loaded in each lane
The text or figure legend describes how RNA was extracted prior to blotting onto the solid support.
The blot has appropriate positive and negative control lanes (positive control might demonstrate hybridization of the probe with itself; negative control might be the unmodified isogenic parent microorganism).
The dot blot system and hybridization protocols are described in the text or in a cited literature reference, and any modifications of the cited protocols are described in the submitted text.
The probe used for the hybridization is adequately described (in the text or in a figure) as to enable interpretation of the results.
The methodology or citation for quantitative analysis (if done) has been provided and a sufficient number of replicates or samples have been tested to determine whether there are significant differences between samples or treatments.
Checklist for western blot data
Checklist Included?
Figure number and title are included.
Lanes are clearly labelled on the blot.

The figure legend describes each lane of the blot, including the following for the protein loaded:

  • the type of material loaded (example, crude, pure, total extract)
  • the source of material loaded
  • the quantity of material loaded
The protein extraction method is adequately described in the text or in the figure.
The antibody or antiserum preparation protocol is adequately described in the text, including an adequate description of the antigen and its purity. The specificity of the antibody or antiserum has been determined and is described in the text or in a cited literature reference.
The gel system and the blotting protocol are adequately described in the text or in a cited literature reference.
The position of the molecular weight standards is indicated, and the standards cover the appropriate range for the proteins expected to be detected on the blot.
The blot includes appropriate positive and negative controls.
A normal serum control was conducted.
Superfluous bands and background signals are properly explained.
The methodology or citation for the quantitative analysis (if done) has been provided, and a sufficient number of replicates or samples have been tested to determine whether there are significant differences between samples or treatments.
Checklist for PCR data
Checklist Included?
The PCR gel has a figure number and title.
The lanes are labelled on the gel.

The figure legend describes each lane of the gel, including a description of the following for the DNA that was loaded:

  • the type of material loaded (example, plasmid fragment, amplified DNA)
  • the source of the material used in each reaction loaded
  • the quantity of material loaded
The position of the molecular weight standards is indicated and covers an appropriate size range for the fragments that are expected to be detected on the gel.
The text or figure legend describes the PCR amplification method performed prior to electrophoresis.
The primers used for amplification are described in the text or in the figure sufficiently, to enable interpretation of the results.
The gel has appropriate positive and negative control lanes (positive control might demonstrate specificity of the primers and the ability to amplify the appropriate size band; negative controls might include amplification with DNA from the unmodified isogenic microorganism and amplification in absence of DNA template).
The plasmid or restriction fragment used as the positive control template, is clearly described in the text or in the figure legend, so as to enable interpretation of the PCR results.
The gel system and the PCR protocol are described in the text or in a cited literature reference, and modifications to the cited protocol (if applicable) are described in the text.
Checklist for ELISA data
Checklist Included?
The table has a number and title
All entries are clearly identified in the table and are described in the text or table legend.
The sample preparation method is described.
The antibody or antiserum preparation protocol is adequately described in the text, as well as the antigen and its purity. The specificity of the antibody or antiserum has been demonstrated and described in the text or in a cited literature reference.
The ELISA protocol used is described in the text or cited in the scientific literature, and any modifications to a cited protocol are described in the text.
The positive controls (example,purified protein) and negative controls (example, normal or preimmune serum, non-transformed material) were used.

When ELISA is being used to quantify protein expression in genetically modified microorganisms:

  • the method for the determination of protein expression by the GMM is presented in the text or in a cited literature reference
  • a standard curve was prepared and the limit of detection indicated
  • a sufficient number of replicates or samples have been tested to determine whether
  • there are significant differences between samples or treatments, and statistical analysis was performed
Checklist for enzyme assays
Checklist Included?
The figure (or table) has a title and a legend.
For graphical representations or tables, the axes or columns are labelled and the units indicated.
The scale of the figure accurately represents the data and allows interpretation of the data.

The legend or text describes:

  • the substrate and amount used for the reaction
  • the quantity and origin of the enzyme
  • the temperature and pH
  • other applicable conditions
The text or legend describes the extraction and purification of the enzyme and the degree of purification achieved.
The assay method and relevant information concerning the enzyme have been provided in the text or in a cited literature reference.
Appropriate controls are included in the assay.
The stability of the enzyme has been taken into account in the design of the assay or the interpretation of the data.
The kinetics of the enzyme have been calculated and, where possible, compared to published data.
If quantitative analysis was performed, a sufficient number of replicates or samples have been tested to determine whether there are significant differences between samples or treatments, and statistical analysis was performed.

Appendix III: Vector description

This information is taken from the document Molecular Genetic Characterization Data, prepared jointly by the CFIA, Health Canada and USDA-APHIS in 1998. It describes the critical elements to be considered during the review of the molecular genetic characterization data of transgenic plants. The same format should be followed for the submission of molecular characterization data of microbial products.

Table 1: Example of a table describing the DNA components of a PV-STBT02 vector (from APHIS petition # 94-257-01p)
Genetic
Element		 SizeTable Note 1
Kb		 Function and Source
RB 0.36 A restriction fragment from the pTiT37 plasmid containing the 24 bp nopaline-type T-DNA right border used to initiate the T-DNA transfer from Agrobacterium tumefaciens to the plant genome (Depicker et al., 1982)
E35S 0.62 The cauliflower mosaic virus (CaMV) promoter (Odell et al., 1985) with the duplicated enhancer region (Kay et al., 1987).
cryIIIA 1.8 The gene which confers resistance to CPB. The gene encodes an amino acid sequence identical to the CPB control protein (referred to as the B.t.t. Band 3 protein) found in B.t.t. as described by Perlak et al. (1993).
E9 3' 0.63 A 3' nontranslated region of the pea ribulose-1,5-bisphosphate carboxylase small subunit (rbcS) E9 gene (Coruzzi et al., 1984), which functions to terminate transcription and direct polyadenylation of the cryIIIA mRNA.
NOS 3' 0.26 A 3' nontranslated region of the nopaline synthase gene which functions to terminate transcription and direct polyadenylation of the nptII mRNA (Depicker et al., 1982; Bevan et al., 1983).
nptII 0.79 The gene isolated from Tn5 (Beck et al., 1982) which encodes for neomycin phosphotransferase type II. Expression of this gene in plant cells confers resistance to kanamycin and serves as a selectable marker for transformation (Fraley et al., 1983).
35S 0.32 The 35S promoter region of the cauliflower mosaic virus (CaMV) (Gardner et al., 1981; Sanders et al., 1987).
LB 0.45 A restriction fragment from the octopine Ti plasmid, pTi15955, containing the 24 bp T-DNA left border used to terminate the transfer of the T-DNA from Agrobacterium tumefaciens to the plant genome (Barker et al., 1983).
ori V 1.3 Origin of replication segment for ABI Agrobacterium derived from the broad-host range plasmid RK2 (Stalker et al., 1981).
ori-322/rop 1.8 A segment of pBR322 which provides the origin of replication for maintenance of the PV-STBT02 plasmid in E. coli, the replication of primer (rop) region and the bom site for the conjugational transfer into the Agrobacterium tumefaciens cells (Bolivar et al., 1977; Sutcliffe, 1978).
aad 0.93 A fragment isolated from transposon Tn7 containing a 0.79 kb gene which encodes for the enzyme streptomycin adenylyltransferase that allows for bacterial selection on spectinomycin or streptomycin (Fling et al., 1985).

Table Notes

Table Note 1

Sizes are approximations.

Return to table note 1  referrer

Figure 1: Example of a detailed map of a plasmid vector
(from APHIS petition #94-257-01p)

Detailed map of a plasmid vector. Description follows.
Description for images – Figure 1. Plasmid map of PV-STBT02.

Restriction sites, and their locations in base pairs, utilized during Southern analyses are shown. The region which served as the T-DNA is marked and it's delineating right and left borders are denoted by open arrows. The blackened regions denote positions of homology for PCR probes used during Southern analyses as described in Section V.A. Cleavage sites for HindIII, EcoRl, Xhot and Notl restriction endonucleases are shown. A description of the genetic elements appears in Table 1.

References

FAO/WHO (2000). Joint FAO/WHO Consultation on Foods Derived from Biotechnology: Safety aspects of genetically modified foods of plant origin. World Health Organization 29 May – 2 June 2000, WHO Headquarters, Geneva, Switzerland

FAO/WHO (2001). Joint FAO/WHO Consultation on Foods Derived from Biotechnology: Evaluation of the allergenicity of genetically modified foods, 22‑25 January 2001, Rome, Italy

OECD (1993). Safety evaluation of foods derived by modern biotechnology: concepts and principles. Organization for Economic Development and Cooperation, Paris.

2.8 Guidance on bridging an application to data from publicly available literature and previously approved feed applications

Data to support the efficacy or safety of a feed can be provided by the applicant from in-house studies, peer-reviewed scientific literature, or in some cases, by reference to previous applications to the Canadian Food Inspection Agency (CFIA). When applicants refer to data other than from complete in-house studies, a "bridge" is required to demonstrate the studies' relevance to the product in the current application.

Bridging, when appropriate, is carried out to support the efficacy and/or safety requirements for the approval and registration of a single ingredient feed (SIF) or registration of mixed feeds. The guidance on information requirements to support the registration/approval of feeds is found in "RG-1 Regulatory Guidance: Feed registration procedures and labelling standards".

The CFIA can consider data from peer-reviewed scientific literature or previous applications only if the bridging requirements below are met:

A. Requirements for bridging publicly available data

Only peer-reviewed scientific literature will be considered acceptable as a source of publicly available data. All information, data and results in the research article will be assessed and considered during the evaluation. The research article does not need to be published by the applicant. However, the study's relevance in the context of the feed under review must be explained in the application, as described below.

For each scientific paper used for bridging data, the applicant must provide a copy of the paper and a summary that includes an explanation of all the relevant information and results. This summary must include substantiation or a rationale:

For example, a beef cattle study may be used to support feed applications for other meat- producing ruminants. However, it may not be relevant to support feed applications for lactating or reproducing ruminants and typically cannot be used to support a feed application for monogastric livestock.

The source of the data from the peer-reviewed scientific literature must be cited in the application (author/source, year), and a list of the complete references must be provided. Each reference must include authors, document title, source, date and page numbers.

B. Requirements for bridging to data in previous applications

Applicants can also bridge data from previously approved applications by referencing their own data that were previously submitted and accepted for approval or registration.

  1. The applicant must include a summary of the data and a scientific rationale explaining why the data from a previous application applies to the ingredient or product under review.
  2. Bridging of data from a previous application will be considered acceptable when the experiments were designed and performed for the same purpose as in the new application or, if not, would still allow conclusions on the feed under application to be made.
  3. The applicant must provide evidence to demonstrate that the previously assessed ingredient or product is equivalent to the ingredient or product under review. For example, the ingredient form (physical and chemical), manufacturing processes and the feed previously assessed use rate must be relevant to the feed being assessed. If not identical, the bridging rationale must be sufficient to still allow conclusions on the feed under application to be made.
  4. For the CFIA to consider applicable data from other applications, the applicant may either:
    • Provide all of the bridged information that would apply to the new application in the current application package as an appendix, or
    • Provide the reference number or feed registration number and original registration date corresponding to the application for which the original data were supplied so the CFIA may efficiently locate it

C. Requirements for bridging to data belonging to a third party company

Where appropriate, an application for a feed ingredient or product may be bridged to data submitted by a third party company that previously received approval or registration for a comparable ingredient or product; or, under certain conditions, for the same ingredient or product included in the new application. Along with the information in Section B, the applicant must:

Checklists

Checklist for bridging to peer-reviewed scientific literature

The checklist below can be used to verify that all aspects of bridging are included in a submission whenever peer-reviewed scientific literature is used to provide information relevant to submission requirements

Checklist 1
Location in submission
(page/file name)		 Check or N/A
1.

Provide reference citation:

Authors. (Year) Title. Journal Title. Volume (Issue): page-page.

Example:

Hyde, M. L., M. R. Wilkens, and D. R. Fraser. (2019) In vivo
measurement of strontium absorption from the rumen of
dairy cows as an index of calcium absorption capacity. Journal of
Dairy Science. 102(6):5699–5705.
2.

A copy of the peer-reviewed scientific publication has been included

  • electronic Portable Document Format (pdf) preferred
3. A summary of the relevant information in the publication related to the data requirement it is intended to support has been provided addressing items 4 to 6 below
4.

Information to demonstrate the equivalence of the ingredient or product used in the study to the product under review.

  • If not identical, the bridging rationale must be sufficient to still allow conclusions on the feed under application to be made
5.

The ingredient or product use rate in the published study must be  compared to that of the ingredient or product under review and must be:

  • equivalent when used to support the efficacy
  • equivalent or higher when used to support the safety
6.

The equivalence of the animal in the study to those intended to be fed the ingredient or product under review in consideration of:

  • species, type, and phase of production

If necessary a rationale for extrapolation to physiologically related species and production purposes

Checklist for bridging to data in previous application or third party information

The checklist below can be used to verify that all aspects of bridging are included in a submission whenever an applicant is referencing data previously submitted and accepted for approval or registration to provide information relevant to submission requirements.

Checklist 2
Bridging rationale elements Location in submission
(page/file name)		 Check or N/A
1. A summary of the previously approved data
2. Include a scientific rationale explaining why the data from a previous application applies to the ingredient or product under review addressing items 3 to 5 below
3.

Information that experiments were designed and performed for the same purpose (including use) as in the new application

  • If not the same, the rationale must be sufficient to still allow conclusions on the feed under application to be made
4.

Provide evidence to demonstrate that the previously assessed ingredient or product is equivalent to the ingredient or product under review

  • If not identical, the bridging rationale must be sufficient to still allow conclusions on the feed under application to be made
5.

The equivalence of animal in the study to those intended to be fed the ingredient or product under review in consideration of:

  • species, type, and phase of production
  • if necessary, rationale extrapolation to physiologically related species and production purposes
6.

All of the bridged information that would apply to the new application in the current application package as an appendix

  • The reference number or feed registration number and original registration date corresponding to the application for which the original data was supplied
7.

If bridging to data from a third party, an attestation from an individual with signing authority from the company holding the registration/approval is provided and indicates:

  • the third party company does not object to having their file and data being used by the CFIA to support data requirements for feed ingredient/product approval/registration for the current application

2.9 Data flexibility for production performance endpoints and associated claims

The CFIA is introducing flexible approaches to the data required to fulfill the efficacy portion of a pre‑market submission. This guidance explains the flexibility being allotted with regards to the studies required to support production performance claims.

Previously, in order to make a specific production performance claim for a feed, an applicant had to provide data to support the claims from at least 3 independent studies per species and production class. Some examples (not an exhaustive list) of specific endpoints for which production performance claims may be made are:

There are now three ways where flexibility may be gained to support these types of claims without compromising the safety of a product.

  1. Combining specific endpoints to make a general production performance claim
  2. Extrapolating data between similar physiological species
  3. Strength of claim – adding more flexibility with the p-value

In all cases, the applicant is responsible for clearly describing to the CFIA how their efficacy data will utilize these flexibilities. This means that the submission package must include a summary detailing any linkages between the studies submitted and the proposed claims. In no case can these flexibilities be considered in place of submitting required safety studies/data.

I. Combining specific endpoints to make a general production performance claim

Consistency in the reporting of, or significance of, specific endpoints across different studies (in-house or published) in relevant species and/or production classes of livestock can make it difficult to meet the 3-study threshold for a specific claim. However, it is now possible to make a general production performance claim. Where at least 3 independent studies demonstrate significant beneficial endpoints, even if they are different, a general production performance claim can be made. Some examples of how specific and non-specific/general claims may be assessed are presented in Table 1.

Table 1: Examples of non-specific and non-specific/general improved production performance claims
Efficacy data provided Claim

An application for a dairy feed supplement

  • 3 studies show increased milk production (p ≤ 0.05) and increased feed intake (p ≤ 0.05)
 Increased milk production and feed intake in dairy cows

An application for a dairy feed supplement

  • 3 studies show increased milk production (p ≤ 0.05)
  • 2 studies only show increased feed intake (p ≤ 0.05)
 Increased milk production in dairy cows

An application for a dairy feed supplement

  • 2 studies show increased milk production (p ≤ 0.05)
  • 1 study only shows increased feed intake (p ≤ 0.05)
 Increased production performance in dairy cows

An application for a growing swine feed supplement

  • 3 studies show increased average daily gain (p ≤ 0.05) and increased feed intake (p ≤ 0.05)
 Increased average daily gain and feed intake in growing swine

An application for a growing swine feed supplement

  • 3 studies show increased average daily gain (p ≤ 0.05)
  • 2 studies showed increased feed intake (p ≤ 0.05)
 Increased average daily gain in growing swine

An application for a growing swine feed supplement

  • 2 studies show increased average daily gain (p ≤ 0.05)
  • 1 study showed increased feed intake (p ≤ 0.05)
 Increased production performance in growing swine

II. Extrapolating data between similar physiological species

An approach for some feed claims where the extrapolation of efficacy data from certain species to other physiologically related species could be used to reduce the total number of studies needed to substantiate a claim for multiple physiologically similar species.

In general, conclusions from studies in animals raised for meat can be extended to include animals of the same species that are reared for reproduction but before reproduction occurs (e.g., from broiler chickens to replacement chickens reared for laying/breeding (pre-layers/replacement hens pre-breeding), from beef cattle to replacement heifers intended for breeding or milking, but before reproduction or milk production occurs). Efficacy data cannot generally be extrapolated between categories of the same species at different production classes (e.g., from broiler chickens to laying or breeding hens).

When a feed application covers several target species in the same production class, it is recognized that it may be unrealistic to expect studies in all potential target species for which an application is made. Therefore, interspecies extrapolation of data can be applied.

In principle, data can be extrapolated between physiologically similar species (Table 2) in the same production class. The degree to which species are physiologically related is judged predominantly in terms of gastrointestinal physiology and function. Similarities in metabolism are also considered. Interspecies extrapolation can be applied in cases where:

  1. the animals are raised for the same purpose, i.e., meat production or reproduction (including milk or egg production)
  2. the product mode of action can reasonably be presumed to be the same between species
  3. the effect(s) claimed is(are) the same
  4. the minimum effective in-feed concentration in the physiologically related species must be the same as that established in the species/category from which data was extrapolated
Table 2: Extrapolation of efficacy data from certain species to other physiologically related species
Major species Characteristics Physiologically related species
Broiler chickens Chickens raised for meat production Other poultry (for example, turkeys, ducks, geese, pheasants, quail, guinea fowl etc.) raised for meat and laying/breeding poultry prior to laying (replacement hens pre-breeding/pre-layers)
Laying hens Productive female birds held for egg production purposes Other laying poultry used for egg production and breeding (for example, turkeys, ducks, geese, pheasants, quail, guinea fowl etc.)
Weaned piglets Young animals having completed the suckling period Other weaned Suidae
Growing pigs Animals intended for meat production until the day of transport to the slaughterhouse Other growing Suidae
Sows Female animals having been inseminated or mated Other reproductive Suidae
Calves Calves which are reared for reproduction, veal production or beef production prior to rumination Other young ruminants prior to rumination (for example, sheep (lambs), goats (kids), buffalo)
Growing cattle Bovine animals that have completed the weaning period with fully developed rumen Other growing ruminants (for example sheep, goat, buffalo)
Dairy cows Lactating or prepartum female bovine Other dairy ruminants (for example, goat, sheep, buffalo) at the corresponding developmental stage
Salmon fish Growing salmonids Other salmonids (for example, trout, arctic char)
Horses Horses kept for performance or meat production Other Equidae
Breeding Mares Mares that have become pregnant at least once Other breeding Equidae
Rabbits Rabbits that are reared for reproduction or meat production Other Leporidae at the corresponding developmental stage at the corresponding developmental stage
Breeding Doe Rabbits Does that have become pregnant at least once Other breeding Leporidae

When the application covers multiple species/classes, the minimum number of independent studies showing the intended effect is shown in Table 3. For applications covering all food-producing animal species, efficacy should be demonstrated in species with different digestive systems. Therefore, studies should be provided to support efficacy for all pigs, all poultry, all ruminants and all fin fish, according to Table 3. Companies can apply to have applications expanded over time to add additional species/classes of livestock as data becomes available.

Table 3: Minimum number of independent studies required for the assessment of efficacy in applications intended to multiple species/categories
Application For Number of studies required and species
All meat and growing poultry species 3 in broiler chickens Table note a
All laying and breeding poultry species 3 in laying chickens Table note a
All poultry species 3 in chickens raised for meat Table note a
+ 3 in laying hens Table note a
All growing pigs 3 in weaned piglets Table note a
+ 3 in growing pigs Table note a
All pigs 3 in weaned piglets Table note a
+ 3 in sows Table note a
All growing ruminants (see below) 3 in calves (pre-ruminants) Table note a
+ 3 in grower cattle Table note a
All ruminants 3 in calves Table note a
+ 3 in cows Table note a
All fin fish 3 in salmonids
+ 3 in (1 each) in other species
All horses 3 covering both growing and reproductive animals
All rabbits 3 covering both growing and reproductive animals

III. Strength of claim – adding more flexibility with the p-value

Historically, the CFIA has followed the standard used by peer-reviewed scientific journals for research when defining the statistical significance p ≤ 0.05 (α=0.05). Considering the following points will allow for flexibility in the significance level and reduced endpoint specificity and allow a predictable strength of claim approach.

Table 4: Strength of claim options
p-value 3 Studies for specific production performance endpoints 3 Studies with different production performance endpoints (see Section I above)
p ≤ 0.05 Improves specific endpoint Improves production performance
0.05 < p ≤ 0.10 May improve specific endpoint May improve production performance
p > 0.10 Not acceptable Not acceptable
Table 5: Examples of strength of claim options
Study overview p-values Claim
3 Studies showing improved feed conversion ratio in beef cattle All studies p ≤ 0.05 Improves feed conversion ratio in beef cattle
3 studies showing increased feed intake in growing and finishing swine All studies p ≤ 0.10 (some could be ≤ 0.05) May increase feed intake in growing and finishing swine
2 studies showing increased daily gain in growing swine and
1 study showing increased feed intake growing swine		 All studies p ≤ 0.05 Improves production performance in growing swine
2 studies showing increased daily gain in beef cattle and 1 study showing increased feed intake in beef cattle All studies p ≤ 0.10 May improve production performance in beef cattle
3 studies showing increased feed intake in laying hens One study p ≤ 0.05
Two studies p ≤ 0.10		 May increase feed intake in laying hens
2 studies showing increased daily gain and
1 study showing increased feed intake in growing swine		 Two studies p ≤ 0.05
One study p ≤ 0.10		 May improve production performance in growing swine
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