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Veterinary Biologics Guideline 3.17E

Guide for Reporting Laboratory and Field Efficacy Trials

INTRODUCTION

These guidelines have been prepared by the Veterinary Biologics Section (VBS) in cooperation with the Biologics Evaluation Laboratory (BEL). Their function is to standardize the submission of efficacy data submitted by a manufacturer in support of a request made in Canada for a Veterinary Biological Product Licence or a Permit to Import a Veterinary Biologic. Because of the large number of biological products imported from the United States, these guidelines take into account the formats used by the Code of Federal Regulations 9 (9CFR) of the United States.

Under the Canadian Health of Animals Act (64.(1)(s)) and regulations, veterinary biological products must be demonstrated by the manufacturer to be efficacious according to the label claim in the target animal species. The efficacy of a veterinary biological product is evaluated using laboratory-based vaccination-challenge trials. Field trials with natural challenge are only used to make licensing decisions, if laboratory-based disease models in the target animal species are not available for study. Products are not released for field efficacy or safety trials without the previous submission and acceptance of the following by VBS: 1) an Outline of Production, 2) a summary of purity, potency and safety test results from the experimental serial(s), and 3) data from a laboratory-based vaccination-challenge efficacy trial.

Protocols for both laboratory and field efficacy trials should follow the format of these guidelines. If the efficacy trial is also to be used in support of product safety, then a description of the responses etc. used to measure safety should be included in the protocol for the efficacy trial.

Copies of all efficacy trial results (trials to select dose, antigen level, adjuvant, definitive or repeated trials) relating to a veterinary biological product submission (whether or not the product was shown to be effective) should be maintained on licensed premises and be available for inspection at all times.

Part I (A-L): Experimental Protocol, describing the proposed trial protocol, together with the Outline of Production should be sent for review to VBS prior to the trial, allowing a maximum of 60 days for review. The trial should not be started until Part I (A-L): Experimental Protocol has been accepted by VBS. VBS may arrange to monitor various parts of the trial. Copies of the purity, potency and laboratory safety results of the experimental serial to be used in the efficacy trial should also be sent to VBS prior to the start of the trial.

Part 11 (M-O): Results and Conclusions, describing the summary, results and conclusion/discussion together with a final copy of the experimental protocol for the trial should be submitted upon completion. Protocols for both laboratory and field efficacy trials should be submitted to:

Veterinary Biologics Section
Canadian Food Inspection Agency
2 Constellation Crescent (Floor 8)
Ottawa, Ontario K1A 0Y9

GENERAL REQUIREMENTS

The submission of the pre-trial efficacy protocol or data from an efficacy trial should be submitted in the following recommended format.

1. The submission should include the following sections. For some products, it may be necessary to include additional headings:

   Part I (A-L): Experimental Protocol
   Title Page, Table of Contents, Participants, Description of Disease and Rationale for Vaccination, References, Objective of the Trial, Experimental Design, Trial Schedule, Description of Test Materials, Description of Test Animals, Target Animal Challenge Model, Data Collection Procedures, Statistical Methods, Tables, Figures and Appendices.

   Part II (M-O): Results and Conclusions
   Title Page, Table of Contents, Summary, Results, Conclusion/Discussion, Tables, Figures, and Appendices.

2. The manufacturer's assigned scientific name and trade name (if known) of the biological product (or component), the establishment licence number, and the date prepared should appear on the title page, and each page of the efficacy trial.

3. Within each Part (I and II) of the efficacy trial, number the pages consecutively, beginning with the title page. Amended pages should be numbered the same as those being superseded. They should bear the date prepared, and the date of the pages being superseded. If one page is superseded by more than one page, this page number should be followed by letters.

CANCELLATION / CHANGES

1. Should changes to the experimental protocol describing the efficacy trial be necessary, the manufacturer should notify VBS in writing, including the following: a) a summary of the necessary changes with reference to each part, page, paragraph, or subparagraph being changed, b) an explanation of the reason for the change(s), and c) a copy of the amended page(s).

2. If the submission is a draft proposal for an efficacy trial, and for some reason the trial does not take place as indicated, contact VBS:

   Veterinary Biologics Section
   Canadian Food Inspection Agency
   2 Constellation Crescent (Floor 8)
   Ottawa, Ontario K1A 0Y9

LABORATORY BIOSAFETY

All aspects of the proposed efficacy trial must meet the standards and regulations for laboratory safety described in the most current edition of the Laboratory Biosafety Guidelines (Health and Welfare Canada and Medical Research Council of Canada), relevant federal, provincial legislation and local safety authorities.

The guidelines do not reference all animal pathogens. However, the manufacturer is responsible for ensuring that the appropriate containment level is used to handle the live organisms used in the efficacy trial.

Laboratory Biosafety Guidelines
or
Material Safety Data Sheets

Office of Biosafety
Laboratory Centre for Disease Control
Public Health Agency of Canada
Ottawa, Ontario K1A OL2 Canada

USE OF TARGET ANIMALS IN THE EFFICACY TRIAL

All aspects involved in the proposed use of animals in the efficacy trial must meet the standards and regulations for the care and maintenance of experimental animals as described by the Canadian Council on Animal Care, relevant provincial legislation and local animal care authorities.

Guide to the Care and Use of Experimental Animals: Volume I and II
Canadian Council on Animal Care
1105 - 151 Slater Street
Ottawa, Ontario K1P SH3 Canada

PART I (A-L): EXPERIMENTAL PROTOCOL

A. TITLE PAGE

The title page should contain the following:

1. Scientific name, and if known, the trade name of the veterinary biological product (or component)
2. Establishment licence number
3. Name of the manufacturer (or foreign manufacturer)
4. Trial title
5. Anticipated start and completion dates of the trial
6. Preparation date of the manuscript
7. Name, address, telephone, fax, and internet numbers of the person to contact regarding the trial
8. Proposed label claim
   (The proposed label claim should be clearly stated. The efficacy trial should be focused toward the support of this specific label claim.)

B. TABLE OF CONTENTS

This section should outline the contents of the manuscript.

C. PARTICIPANTS

This page should list the names, addresses, phone numbers, fax numbers, internet addresses, and the responsibilities of the principle participants in the trial. It should also include a list of the collaborators (producers, veterinarians, etc.) participating in the trial, if this is relevant. The principle participants should confirm their participation. The qualifications and experience of the key personnel should be outlined in Appendix I.

D. DESCRIPTION OF DISEASE AND RATIONALE FOR VACCINATION

Summarize the scientific medical rationale for using the product under review to control or prevent a disease in the target species. This should be supported by publications in the scientific literature, and should include a brief description of the current knowledge of the disease (clinical signs, pathogenesis, epidemiology). Give only key references, and do not review the subject extensively. Appendix II should contain copies of the key papers.

E. OBJECTIVE OF THE TRIAL

The objective of an efficacy trial is to demonstrate that a veterinary biological product is efficacious in the target animal species, according to the label claim made by the manufacturer. Efficacy data should be established for each clinical form of a disease, each route of administration (intramuscular, subcutaneous, intranasal, etc.), and each species (bovine, porcine, canine, feline, equine, etc.) recommended in the label claim. The efficacy trial should be conducted in fully susceptible animals of the youngest age for which the product is recommended. If the youngest age recommended for vaccination is still expected to have levels of maternal antibody, then the efficacy of the product should be established in the face of the expected levels of maternal antibody. If this data is not available, labelling should indicate that the product is for the vaccination of susceptible animals of the minimum age used in the efficacy trial, and recommend re-vaccination at appropriate intervals until such animals reach an age when interfering levels of maternal antibody would no longer be present.

Specific label claims concerning the onset of immunity should also be supported by acceptable efficacy data.

When recommending vaccination of adults to provide passive immunization of offspring, the efficacy must be established in the offspring of a significant number of vaccinated adults.

Serological data are generally not acceptable for establishing the efficacy of a product. They should only be used when reasonable data exists to demonstrate that the serological results are indicative of protection.

The objectives of the trial should be described in detail. This should include: 1) a general overview, and 2) a specific description of the efficacy objectives in terms of the experimental response(s) to be measured, the times of measurement, and the methods. For many products, these are listed in the Code of Federal Regulations 9 (9CFR) of the United States. For information on products not listed here, the specific clinical signs, pathologic, serologic, and/or immunologic responses required for licensing, as well as, the time periods over which they must be measured can be obtained from VBS veterinary reviewers for each product.

For example:

General Objective:
To demonstrate the efficacy / immunogenicity of an infectious bovine rhinotracheitis vaccine, killed virus.

Specific Objective:
In vaccinate and control animals to describe the daily changes in clinical signs of respiratory disease and body temperature on each of 14 days following challenge with virulent infectious bovine rhinotracheitis virus.

If the efficacy trial is also to be used in support of the product safety, then this section should also include a detailed description of the safety objectives of the trial, including: 1) a general overview, and 2) a specific description of the objectives in terms of the experimental response(s) to be measured, the times of measurement, and the methods.

F. EXPERIMENTAL DESIGN

This section should include the following:

1. General Overview of the Experimental Design
   Give a general outline of the trial, describing the treatments, test animals, sampling, timing of events, etc.

2. Allocation of Animals to Treatment Groups
   This section should include: a description of the treatment groups (including any stratification or blocking factors used in the experimental design, such as: antibody titre, age, sex, weight, litter, etc.), their sample sizes, the alpha and power of the trial, the method used to randomly allocate the experimental animals to the treatment groups, and the randomization plan for the trial. The statistical formulae used to estimate the sample sizes should be referenced. For many products the method of allocating animals to treatment groups are specified in the 9CFR.

3. Response Variables and Scoring Method
   For many products, the response to be measured, and the definition of a positive response are outlined in the 9CFR. For products in which this is not the case, this section should describe the biological responses and the scoring systems that are to be used to measure the efficacy of the product, including the range of values to be considered a positive or negative response.

   If the manufacturer decides to use a composite score or index of clinical or pathological signs to demonstrate a significant difference between the vaccinate and control animals, they should also describe and statistically evaluate each response making up the score or index, separately.

   If the efficacy trial is also to be used in support of product safety, then this section should describe the biological responses and scoring systems being used to measure the safety of the product. As with the response variables for efficacy, this should include the range of values to be considered a positive or negative response.

4. Blinding the Trial
   This section should describe the methods used to blind the trial in order to ensure unbiased results. It should include a list of the various aspects of the trial which are blinded and the personnel who will be blinded.

G. TRIAL SCHEDULE

The trial schedule should list the key experimental days, their corresponding dates, and the event(s) which take place (arrival, acclimatization, vaccination, challenge, sampling, observations, etc.).

H. DESCRIPTION OF THE TEST MATERIALS

The composition of the veterinary biological product used in the pre-license laboratory or field efficacy trial should be that of the product intended for sale. It should be one of three experimental serials produced in a licensed facility in accordance with a previously filed Outline of Production. If the experimental serials are produced on a smaller scale in a research facility, the manufacturer must establish that they represent the product which will be produced for sale in the licensed production facilities. The manufacturer is responsible for establishing the validity of the experimental serial used to demonstrate the efficacy of a product by submitting to VBS, the summary test results of the serial for purity, potency and laboratory safety as described in the Outline of Production.

1. Test Materials

   For both the experimental serial and the placebo used in the efficacy trial, describe the composition, including antigenic mass and methods of production, or cite the Outline of Production. This should also include: the date of production, date of use, manufacturer's trade name, manufacturer's scientific product name, serial number, the method of use (dose, route, and frequency) in the efficacy trial and, the purity, potency and laboratory safety results.

   Describe the method by which the level of potency of the experimental serial to be used in the efficacy trial will be measured. This should be the same as that described for serial release in the Outline of Production.

   The experimental serial used in the efficacy trial should be at a potency level equal to that proposed in the Outline of Production at the release of the product for sale.

   The experimental serial used in the efficacy trial should be produced at the highest passage from the Master Seed, and if of cell culture origin, in cells at the highest passage from the Master Cell Stock allowed by the Outline of Production.

2. Use and Disposal Of Unused Portion of an Experimental Serial

   For the experimental serial used in the efficacy trial, identify and account for all material prepared, used, distributed and returned.

   Describe the method proposed to dispose of the unused veterinary biological experimental serial and placebo.

I. DESCRIPTION OF THE TEST ANIMALS

This section should describe the test animals to be used in the efficacy trial. It should include the following:

1. Selection/Exclusion Criteria
   Describe the source, age, breed, sex and any other distinguishing features of the animals used in the trial.

2. Method of Animal Identification
   Describe the method of animal identification.

3. Housing and Environmental Conditions
   This section should include a description of when treatment groups are in contact with one another, or kept separate.

4. Concurrent Medication/Vaccination
   Describe any concurrent medications, vaccinations that will be received by the animals used in the efficacy trial. This should include anaesthetics, analgesics, immobilizing agents and tranquillizers, etc.

5. Treatment of Adverse Reactions
   Describe anticipated adverse reactions and the methods that will be used to treat animals experiencing these reactions.

6. Disposition of study animals and their milk (This should indicate the withdrawal times where applicable.)
   Manufacturers are required to notify VBS in writing, at least two weeks prior to the shipment of experimental animals to slaughter so that the regional veterinarian and the inspection staff of the establishment can be informed.

The following information must be provided: the name and intended use of the experimental veterinary biological product; the name and address of the investigator responsible for the study; the estimated dates of the commencement and completion of the trial; the date of treatment or vaccination; the number of animals and their identification; and the name and address of the federally inspected slaughter house where animals would be sent for slaughter.

Note that for meat animals a withdrawal time of at least 21 (or more depending on the type of adjuvant)days is required for veterinary biological products before the animals can be shipped for slaughter. Some of the immobilizing agents used for animal restraint in the different species may have longer withdrawal times.

J. TARGET ANIMAL CHALLENGE MODEL

Summarize the scientific rationale for using this target animal challenge model to support the product efficacy. This should be supported by publications in the scientific literature, and should include a brief description of the current knowledge of the challenge model.

Provide a detailed description of the materials and methods of the challenge, as well as, information on its development, and repeatability. Specifically define a successful challenge in terms of the efficacy response(s) to be measured.

K. DATA COLLECTION PROCEDURES

1. Sample Collection
   This should include a description of the methods used to collect the biological samples for measuring each response (blood, serum, post mortem etc.).

2. Sample Records/Forms
   Samples of the record forms should be included in Appendix III.

3. Record Keeping
   

   a)	should include the initials or the signature of the person responsible for an action.
   b)	no abbreviations or acronyms unless defined.
   c)	properly corrected recording errors (single line cross-out with correct result and initials of person responsible for the correction.

L. STATISTICAL METHODS

This section should outline the strategy that will be used to carry out the statistical analysis to determine the efficacy and/or safety of a product. The analysis should reflect the experimental design, including the randomization scheme and the type of outcome variables measured. It should include a description of the statistical tests to be used, the groups to be compared, the response variables to be used in the comparisons, a statement of whether the scores are to be analyzed as ordinal categories or continuous variables, and the time periods over which the comparisons will be made.

The statistical methods and computer software being used to carry out the calculations should be should be listed and/or referenced.

PART II (M-O): RESULTS AND CONCLUSIONS

Part II should include a separate title page and table of contents. If the efficacy trial is also to be used in support of product safety, then, the data for safety objectives of the trial should be reported in a separate Part III using the same sections and reporting format as described below.

M. SUMMARY

The summary should describe the purpose of the efficacy trial, the basic procedures used and the main findings (specific data and their statistical significance). The main findings should be linked to the proposed label claim.

N. RESULTS

The results should include:

1. summary tables of individual animal data, sorted by treatment group. All subjects entering the trial should be accounted for in the report, as well as in the written records. The original of the data collection record form(s) used in the trial should be kept on file by the manufacturer and be available upon request.

2. descriptive statistics showing the measures of central tendency (means, medians etc.), and the distributions (tables, histograms, bar charts, scattergrams, boxplot etc.) for the efficacy response variables by relevant sub-group.

3. inferential statistics (ANOVA, Mantel-Haenszel etc.) demonstrating the statistically significance difference between the control and vaccinated groups for the efficacy response variables.

O. CONCLUSION/DISCUSSION

The discussion should emphasize the important aspects of the efficacy trial, and the conclusions that follow from them. Include the implications of the findings and their limitations. Link the conclusions with the proposed label claim and objectives of the efficacy trial, but avoid unqualified statements and conclusions not completely supported by the efficacy data. Do not allude to work that has not been completed.