PROTOCOL TITLE:
Evaluation of Inactivated Avian Influenza Vaccines in Zoo Birds
VS Codes: 1057.R3
Date
April 12, 2006
U.S. Veterinary License No. 112
Fort Dodge Animal Health
Fort Dodge , Iowa
USA
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CONTENTS:
1.0
STATEMENTS OF APPROVAL AND COMPLIANCE
2.0
GENERAL INFORMATION
2.1
Title
2.2
Protocol No.
2.3
Study Support
3.0
INTRODUCTION
3.1
Background
3.2
Objective(s)
4.0
MATERIALS AND METHODS
4.1
Test Animals
4.2
Test Vaccines
4.3
Experimental Design
4.4
Vaccination
4.5
Observation procedure
4.6
Sample Collection and Testing
4.7
Data Analysis
5.0
DATA INTERPRETATION
B703-06-003.P
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1.0
B703-06-003.P
STATEMENTS OF APPROVAL AND COMPLIANCE
We have reviewed this protocol and agree that it accurately reflects the proposed study,
Evaluation of Inactivated Avian Influenza Vaccines in Zoo Birds. Amendments or deviations to
this protocol will be approved in writing by the undersigned and documented with
explanations(s) in the final report as protocol amendments or deviations.
Jeff Rodenberg, B.Sc.
Manager, Poultry Viral R&D
Fort Dodge Animal Health
________________________________________________
Mahesh Kumar, Ph.D.
Senior Director, Global Poultry R&D
Fort Dodge Animal Health
________________________________________________
Dr. Gary Riggs, D.V.M.
Animal Health Committee
American Zoo and Aquarium Association
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2.0
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GENERAL INFORMATION
2.1
Title
Evaluation of Inactivated Avian Influenza Vaccines in Zoo Birds
2.2
Protocol No. B703-06-003.P
2.3
Study Support
2.3.1
Study Investigator
Gary Riggs, DVM
Animal Health Committee
American Zoo and Aquarium Association
For a list of participating institutions and associated on-site responsible
parties, See Attachment A
2.3.2
Study Co-Investigator
Jeff Rodenberg
Manager, Poultry Viral R&D
Fort Dodge Animal Health
800 5th Street NW
Fort Dodge, IA 50501
2.3.3
Study Sponsor
Mahesh Kumar
Senior Director, Global Poultry R&D
Fort Dodge Animal Health
800 5th Street NW
Fort Dodge, Iowa 50501
2.3.3
Laboratory Support
Jason Hansen, Dave Lutz
Fort Dodge Animal Health
800 5th Street NW
Fort Dodge, Iowa 50501
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3.0
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INTRODUCTION
3.1
Background
Avian Influenza, high-path subtype H5N1 has had a devastating effect on poultry
production in SE Asia. Millions of birds have succumbed to disease, and millions
more have been eradicated in an effort to control the outbreak. Additionally, fatal
cases of human influenza have been linked to the H5N1 virus, raising the
possibility of a virus capable of crossing the species barrier and causing a human
epidemic or pandemic.
The H5N1 virus has been rapidly expanding in geographical distribution, with
recent reports of infected waterfowl and/or poulty in several countries including
France, Germany, Nigeria, Belgium, Poland and Hungary. Some believe it
inevitable that the virus will eventually spread to the United States, thus placing
US poultry and other avian species including those held in zoological facilities at
risk.
A reassortant influenza vaccine candidate has been prepared at St Jude Children’s
research hospital by the method of Hoffman, et al as desribed in Ming Liu et al.
Briefly, a reverse genetics approach was used in which the hemagglutinin (HA)
gene from A/chicken/Vietnam/c58/04 (H5N1), modified to delete the basic amino
acid motif which specifies highly pathogenic virus, was combined with the
neuraminidase gene from A/Duck/Germany/1215/73 (H2N3) and internal genes
from A/Puerto Rico/8/34 (PR8 strain). The result is a H5N3 virus of low
pathogenicity, capable of yielding high titer when propagated in embryonated
chicken eggs, with a high degree of homology to the H5 asian isolate yet still
distinguishable from that isolate by virtue of the heterologous neuraminidase.
Development of this virus into an inactivated vaccine by Fort Dodge Animal
Health is nearing completion, and in fact the vaccine has been approved for use on
an emergency basis in France. Data collected in support of licensure indicates a
high level of protection is elicited by the vaccine, and may include the ability to
prevent infection.
Philippa, et. al.1, reported on a study conducted in The Netherlands in which
several taxonomic orders of avian species kept in Dutch zoos were administered
an inactivated, water-in-oil adjuvanted H7N1 vaccine during the 2003 outbreak of
highly pathogenic H7N7. Vaccine quantity was administered with consideration to
average body weight of the order, and 2 administrations were given with an
interval of approximately 6 weeks between. Serum samples were collected prior
to vaccination and 30-60 days after vaccination for evaluation of serologic
response. The authors found that some orders seemed to respond better than
others, and suggested a smaller interval between vaccinations might elicit a better
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response. Of note, there were no mortalities due to adverse vaccine reactions,
although no information was presented on lesser reactions if they did occur.
The current study, to be conducted in cooperation with the American Zoo and
Aquarium Association (AZA) through its member institutions, seeks to evaluate
the utility of the FDAH reverse genetics H5 subtype vaccine in a similar study in
several avian species kept by zoos. In this study the choice among two
formulations will be made at the discretion of the attending veterinarians at each
institution in conjunction with the Animal Health Committee of the AZA,
inasmuch as they would be most qualified to make a selection appropriate for the
specific bird species. Particular attention will be paid to safety of the vaccines for
parenteral administration and the development of serologic response in vaccinated
birds.
3.2
Objective(s)
Evaluate the safety of and serologic response to inactivated avian influenza
vaccines in various types of zoo birds.
4.0
MATERIALS AND METHODS
4.1
Test Animals
4.1.1
Animal Selection
Species: Multiple
Number: To be determined
Inclusion Criteria: Only healthy birds at time of vaccination will be
enrolled in the study.
Age: Multiple
Source: Multiple
4.1.2
Housing and Care of Animals
Per standard care and husbandry procedures of the cooperating zoological
institutions.
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4.2
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Test Vaccine
4.2.1
Composition of Vaccines
Two vaccines will be prepared for use in the study as follows:
1. Avian Influenza Vaccine, H5N3 subtype (reverse genetics), killed
virus: VS Code 1057.R3
2. Non-mineral oil adjuvanted, H5N3 subtype (reverse genetics), killed
virus. Exact adjuvant formulation to be determined. To be used in
birds known or suspected to be highly sensitive to mineral oil
adjuvants.
Testing records of each vaccine serial will be part of the 9CFR 103.3
request to USDA and will become part of the final report.
4.2.2
Source of Vaccine
Fort Dodge Animal Health, Bio R&D or Fort Dodge Animal Health Bio
Manufacturing.
4.2.3
Shipment of Vaccine
The Animal Health committee in conjunction with the specified attending
veterinarians participating in the study will provide the details of the
amount and type of vaccine to be administered at each institution. Vaccine
will be shipped on cold pack by appropriate courier to maintain a
refrigerated state. No shipment will occur without authorization from the
USDA and State Veterinarian for the particular participating institution.
4.2.4
Serial Numbers
A list of the vaccine serial(s) administered to birds will be recorded and
included in the final report.
4.2.5
Storage
Vaccine formulations will be stored refrigerated at 2-7 degrees C.
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4.3
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Experimental Design
A list of the initial cooperating institutions with responsible personnel is provided
in Appendix A along with a list of the species and approximate number of birds to
be enrolled in those particular institutions. (Note: depending on events in the
field, and for inclusion of species not currently represented, additional cooperators
may be added at a later date by submission of similar detail and State Veterinarian
approval prior to shipment of vaccine).
In brief, birds as selected for participation by the cooperating institutions will be
vaccinated with inactivated avian influenza vaccine, and if possible where
multiple individuals of the same species are enrolled, will include one of those
individuals as a nonvaccinated control. Vaccinations will be given with volume
dependent on consideration of approximate bird weight. Health records as
normally kept by the participating institution will be used in assessing safety, and
bleeding of the birds will be done to evaluate serologic response. It is anticipated
that the data collection will be completed within 6-12 months of the last
vaccination date.
4.4
Vaccination
Vaccine will be administered subcutaneously or intramuscularly in a volume
considered appropriate to the species and in consideration of the age and size of
the bird at vaccination. In general, for birds of less than 1.5 kg, a dose of 0.25 mL
will be injected. For birds of >1.5 kg, a dose of 0.5 mL will be injected. Extremely
large birds such as ratites may be injected with up to 1.0 mL.
Selection of the appropriate vaccination regimen will be at the discretion of the
cooperating institutions in consultation with the Animal Health Committee of the
AZA and/or Study Investigator(s) as necessary, with records kept of the regimen
applied. The vaccination regimen selected will be based on consideration of type
and size of bird, type of adjuvant system employed for the vaccine being
administered, and information on safety/serologic response to previous
vaccinations if available. In general, three different vaccination regimens are
envisioned:
1.) Single vaccination.
2.) Primary vaccination followed by a single booster vaccination at 2 or 4 weeks
post primary injection.
3.) Primary vaccination followed by two booster vaccinations at two week
intervals.
Methods utilized for the restraint and if necessary, anesthesia of birds for
vaccination and/or bleeding will be governed by and the responsibility of the
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cooperating institutions. The administration of anesthesia (if any) will be recorded
also.
Vaccinated animals must be individually and permanently identifiable and the
identity records of these animals must be clearly annotated to indicate the animal
has been vaccinated for avian influenza. If the bird is traded or moved between
zoos, such records are retained with the animal in transport and remain with the
animal for the duration of its life.
A record will be kept of the number of doses administered and volume
administered per dose by the designated responsible party for each cooperating
institution. Upon completion of the vaccination schedule, remaining vaccine will
be returned to the sponsor with documentation reconciling the amount of vaccine
used with the amount of vaccine returned.
4.5
Observation Procedure
Records of pre- and post-vaccination bird weight, regular physical exam findings
and adverse event logs as are normally kept by the cooperating institutions will be
made in accordance with the standard procedures of those institutions.
During the 3 weeks immediately following vaccination, examination of the
injection site will be made inasmuch as it is possible without negative effect to the
health and well-being of the birds.
At three and six months after the booster vaccination, copies of the records for
enrolled birds will be provided to the Study Investigator and Study Sponsor for
use in reporting to state and federal regulatory authorities as may be required.
4.6
Sample Collection and Testing
Blood will be obtained as feasible from the birds for determination of antibody
response. Ideally this would include samples taken at or prior to the first
vaccination, at booster vaccination, and at 6 and 12 months after the last
vaccination. Serum or plasma will be aseptically transferred into separate tubes
and the samples will be frozen at –20 degrees C or colder pending analysis.
Serum/plasma samples will be tested by the Study Sponsor for the presence of
influenza antibody by hemagglutination inhibition assay. Results of antibody
assays will be provided upon completion to the designated responsible individuals
of the cooperating institutions.
ATTACHMENT A: LIST OF PARTICIPATING INSTITUTIONS, ASSOCIATED
CONTACT PERSONNEL, AND ANTICIPATED NUMBERS AND SPECIES OF BIRDS
TO BE VACCINATED