Annex

NOTE: Veterinary certificates are negotiated directly between relevant government authorities. In cases where a veterinary certificate has not been negotiated, this annex can be used as the basis for developing a suitable document.

1. Disease freedom
   At the time of, and for 30 days after, each collection for this consignment, Canada was officially recognised by the World Organisation for Animal Health (OIE) as a:
   1.1. foot and mouth disease free country where vaccination is not practised.
   1.2. contagious bovine pleuropneumonia country free
   AND Canada meets the OIE Animal Health Code Article definitions for country freedom from:
   1.3. Lumpy skin disease,
   1.4. Rift Valley fever,
   1.5. brucellosis due to B. melitensis,
   AND Canada is recognised by the Australian Government as a country free from foot and mouth disease where vaccination is not practised:
2. Vesicular Stomatitis
   During the 30 days prior to, and at the time of, each collection of oocytes, there were no clinical signs or reports of vesicular stomatitis at the premises where donor cows were kept and at the oocyte collection facility.
3. Bovine Brucellosis
   3.1. During the 30 days prior to, and at the time of, each collection of oocytes, the donor cows were from a herd that was located in a country or zone certified free by the CFIA from infection with bovine brucellosis (B. abortus); and
   3.2. infection with Brucella abortus in animals is a notifiable disease in Canada; and
   3.3. no case has been recorded in bovids for at least the past three years; and
   3.4. regulatory measures have been implemented for the early detection of infection with Brucella abortus in bovids; and
   3.5. no bovids have been vaccinated against infection with Brucella abortus for at least the past three years; and
   3.6. bovids and their genetic materials introduced into the country or zone comply with the recommendations in the OIE Animal Health Code.
4. Bovine Tuberculosis
   4.1. During the 30 days prior to, and at the time of, each collection of oocytes, the donor cows were from a herd that was located in a country or zone certified free by the CFIA from bovine tuberculosis; and
   4.2. infection with bovine tuberculosis in animals is a notifiable disease in Canada; and
   4.3. a surveillance programme in accordance with Chapter 1.4. of the OIE Animal Health Code is in place to detect infection with bovine tuberculosis in the country or zone through ante- and post-mortem inspections of bovids; and
   4.4. regulatory measures have been implemented for the early detection of infection with bovine tuberculosis in bovids; and
   4.5. bovids and their genetic materials introduced into the country or zone comply with the recommendations in the OIE Animal Health Code.
5. Schmallenberg Virus
   No cases of disease caused by Schmallenberg virus have been detected or reported in Canada.

6. Embryo production team(s) and team veterinarian(s)
   Note: The embryo production team is a group of competent veterinarians and technicians and includes the Team Veterinarian, to perform the collection and processing of ovaries/oocytes and the production and storage of in vitro produced embryos.
   6.1. The embryo production Team Veterinarian or the embryo production team was approved by the CFIA for export of bovine in vitro produced embryos.

   6.2. The Team Veterinarian is:
   i) certified by the Canadian Embryo Transfer Association as a competent embryo transfer practitioner.
   ii) deemed to be competent in the production of in vitro produced embryos.
   6.3. The embryo production team was supervised by the Team Veterinarian.
   6.4. The Team Veterinarian was responsible for all team operations which include the hygienic collection of oocytes and all other procedures involved in the production of embryos intended for international movement.
   6.5. The embryo production team personnel were adequately trained in the techniques and principles of disease control. High standards of hygiene were practised to preclude the introduction of infection.
   6.6. The embryo production team had adequate facilities and equipment for:
   i) collecting ovaries and/or oocytes
   ii) processing of oocytes and production of embryos at a permanent or mobile laboratory
   iii) storing oocytes and/or embryos.
   These facilities need not necessarily be at the same location.
   6.7. The embryo production team keeps records of its activities, which must be maintained for inspection by the CFIA for a period of at least two years after the embryos have been exported.

7. Oocyte collection facility
   Note: The oocyte collection facility is the premises consisting of an oocyte recovery area and a permanent or mobile laboratory for the processing of oocytes and in vitro maturation before transporting to the in vitro produced embryo processing laboratory. The premises may also include the in vitro produced embryo processing laboratory. The oocyte recovery area is the area dedicated to the ultrasonographically guided aspiration of oocytes and includes facilities for the safe handling of donor cows.
   7.1. The oocyte collection facility:
   i) was on a property not subject to any restriction or quarantine measure with respect to contagious and infectious animal diseases
   ii) was under the supervision of the Team Veterinarian
   iii) was built and maintained in accordance with the recommendations in the current International Embryo Technology Society (IETS) Manual to permit the sanitary collection, handling and processing of the oocytes for maturing
   iv) was subjected to, and passed, inspection at least once a year by an approved veterinarian in the Embryo Production Team
   v) was subjected to review by the CFIA confirming approval at least once a year.
   7.2. Only animals associated with oocyte collection and meeting health requirements as specified in this health certificate were permitted to enter the oocyte recovery area during collection of oocytes for processing to in vitro produced embryos for export to Australia.
8. Oocyte donors
   8.1. Only live animals permanently identified according to an identification system recognised by the CFIA were used for oocyte collection.
   8.2. To the knowledge of the Team Veterinarian, donors showed no clinical signs of contagious and infectious diseases for 30 days prior to, at the time of, and for 30 days after, each collection.
   8.3. The Team Veterinarian or another veterinarian authorised by the Team Veterinarian inspected each female donor on each day that the oocytes were collected for this consignment and certified the donor to be free from clinical signs of contagious and infectious diseases.
   8.4. Donors resided in Canada for at least 90 days prior to oocyte collection for this consignment.
9. Oocyte collection, processing and in vitro maturation
   Note: An oocyte collection is defined as oocytes collected during a single ovum pickup from a live donor.
   9.1. Only oocytes from the same female donor were washed and processed together.
   9.2. All equipment/materials were disposed of and replaced with new items, or sterilised or disinfected in accordance with the current IETS Manual, before use and between different donors.
   9.3. No oocytes of a lesser health status were processed within the laboratory in the same batch as the germplasm for this consignment.
   9.4. Any biological product of animal origin, including media constituents, used in oocyte recovery, maturation, washing and storage presented no animal disease risk. Media were sterilised prior to use by approved methods in accordance with the current IETS Manual and handled in such a manner as to ensure that sterility was maintained. Antibiotics were added to all fluids and media as recommended in the current IETS Manual.
10. Transport of oocytes from oocyte collection facility
    10.1. The oocytes were processed, stored and transported to the in vitro produced embryo processing laboratory in a hygienic manner in accordance with recommendations of the current IETS Manual.
    10.2. Only oocytes from the same individual donor were stored together in the same ampoule, vial or straw.
    10.3. Ampoules, vials or straws were capped or sealed before transport.
    10.4. Where a third party was used for transport, the storage container was sealed at the oocyte collection facility by the Team Veterinarian or a competent veterinarian who is a member of the embryo production team and the seal was not broken until receipt by the Team Veterinarian or a member of the embryo production team at the in vitro produced embryo processing laboratory.
11. Semen
    11.1. Only semen certifiable for export to Australia was used to fertilise the oocytes. Documentation was provided to the CFIA for verification.
    11.2. Canadian origin semen comes from a semen donor who has been resident in Canada for 90 days prior to the collection of semen used to fertilise the oocytes in this consignment,
    11.3. If the semen was collected in another country, the semen importer provided a copy of certification from the country of origin to the CFIA as evidence that the semen met Australian import requirements.
12. In vitro produced embryo processing laboratory
    Note: The in vitro produced embryo processing laboratory is the facility at which the in vitro produced embryos were processed through, at minimum, in vitro fertilisation, in vitro culture, embryo washing and freezing.
    This facility, at the time of embryo processing:
    12.1. was on a property not subject to any restriction or quarantine measure with respect to contagious or infectious animal disease
    12.2. was under the supervision of the Team Veterinarian
    12.3. is a permanent structure that was built and maintained in accordance with the recommendations of the current IETS Manual
    12.4. was subjected to, and passed, inspection at least once a year by the Team Veterinarian
    12.5. was subjected to review confirming approval at least once a year by the CFIA.
13. Production and storage of embryos
    13.1. During the production of embryos for export to Australia and prior to their storage, no oocytes or embryos of a lesser health status were processed at the same time using the same equipment and materials.
    13.2. All equipment/materials were disposed of and replaced with new items, or disinfected in accordance with the recommendations of the current IETS Manual between different donors.
    13.3. Any biological product of animal origin, including co-culture cells and media constituents, used in fertilisation, culture, washing and storage presented no animal disease risk. Media were sterilised prior to use by approved methods in accordance with the current IETS Manual and handled in such a manner as to ensure that sterility is maintained. Antibiotics were added to all fluids and media as recommended in the current IETS Manual.
    13.4. Cleaning, and sterilisation or disinfection of, equipment were carried out in accordance with the recommendations of the current IETS Manual.
14. Embryos
    The embryos were handled in accordance with the current IETS Manual:
    14.1. All embryos are identified and can be traced to the male and female donors.
    14.2. Only embryos from the same female donor were washed together, and no more than ten embryos were washed at any one time.
    14.3. The zona pellucida of each embryo, before washing, was examined over its entire surface area at not less than 50X magnification to ensure that it is intact and free of adherent material.
    14.4. The embryos were washed at least ten times with at least 100–fold dilutions between each wash, and a new sterile micropipette was used for transferring the embryos through each wash.
    14.5. The standard washing procedure includes additional washes with the enzyme trypsin.
    14.6. Eligible embryos include those subject to micromanipulation only for the purpose of collecting biopsy samples for genetic analysis. No other micromanipulations such as splitting, transgene injection, intracytoplasmic sperm injection, nuclear transfer or other interventions that breach the integrity of the zona pellucida of embryos are permitted. If performed, micromanipulation was carried out only on embryos with intact zona pellucida after the standard washing procedure, and in suitable laminar-flow facilities which were properly cleaned and disinfected between batches.
15. Diagnostic testing
    15.1. The samples were collected by veterinarians approved by the CFIA for export certification.
    15.2. Tests for disease were carried out at a laboratory approved by the competent authority to perform the required test.
    15.3. The tests were conducted in accordance with the current OIE Manual.
    15.4. The test reports provided to CFIA to support certification must display the dates of sampling for the tests required, the type of test used and the test results. This information is contained in a table against donor information, annexed to the health certificate, and verified by the CFIA certifying officer.
16. Bluetongue (BTV)
    Blood samples drawn from each donor were either:
    16.1.
    i) subjected to a cELISA test to detect antibodies to the BTV group between 28 and 60 days after each collection of oocytes for this consignment with negative results, or
    ii) subjected to an agent identification test on a blood sample taken on the day of oocyte collection with negative results.
    OR
    16.2. All donors were kept in a country that was free, or seasonally free, from BTV as recognised by Australia* at least 60 days prior to, and at the time of, collection of the oocytes.
    (*Australia recognises Canada as a country seasonally free from BTV without testing between January 1st and May 15th, excluding the Okanagan Valley of British Columbia.)
    (Delete unused option; manual deletion must be initialled by the Official Government Veterinarian)
17. Epizootic haemorrhagic disease (EDHV)
    Blood samples drawn from each donor were either:
    17.1.
    i) subjected to a cELISA test to detect antibodies to the EHDV group between 28 and 60 days after each collection of oocytes for this consignment with negative results, or
    ii) subjected to an agent identification test on a blood sample taken on the day of oocyte collection with negative results.
    OR
    17.2. All donors were kept in a country that was free, or seasonally free, from EHDV as recognised by Australia* at least 60 days prior to, and at the time of, collection of the oocytes.
    (*Australia recognises Canada as a country seasonally free from EHDV without testing between January 1st and May 15th, excluding the Okanagan Valley of British Columbia.)
    (Delete unused option; manual deletion must be initialled by the Official Government Veterinarian)
18. Infectious bovine rhinotracheitis (IBR) and infectious pustular vulvovaginitis (IPV)
    For all oocytes collected from the donors, either:
    18.1.
    i) The donor came from an IBR/IPV free herd as defined in the current OIE Code,
    OR
    ii) The donor was subjected, with negative results, to a serological test for IBR/IPV on blood samples collected at least four weeks after each oocyte collection
    OR
    18.2.
    i) The donor was kept in a herd where all eligible animals including the donor were vaccinated against IBR/IPV with a vaccine approved by the CFIA/Health Canada at least 30 days prior to collection of oocytes. The vaccine was administered as per manufacturer’s instructions for vaccination and revaccination,
    AND
    ii) A vaginal mucus sample* was collected from the donor immediately prior to washing/disinfecting the vagina in preparation for oocyte collection and tested for bovine herpesvirus-1 by the qRT-PCR for bovine herpesvirus-1 with negative results.
    (*Double guarded culture swabs, appropriately prepared for testing samples, were used to hygienically collect samples of vaginal mucus from the donors.)
    (Delete unused option; manual deletion must be initialled by the Official Government Veterinarian)
19. Bovine viral diarrhoea virus (BVDV)
    19.1. At the time of each collection of oocytes, each female donor gave a negative result to one of the following tests for BVDV:
    i) an antigen-capture ELISA on peripheral blood leucocytes,
    OR
    ii) a monoclonal immunoperoxidase or other virus isolation test on blood or serum.
    AND
    19.2. If vaccinated, the donors were kept in a herd where all eligible animals including the donors were vaccinated against both BVDV-1 and BVDV-2 with a vaccine approved by the CFIA / Health Canada at least 30 days prior to collection of oocytes. The vaccine was administered as per manufacturer’s instructions for vaccination and revaccination.
    (Delete unused option; manual deletion must be initialled by the Official Government Veterinarian)
20. Storage and transport
    20.1. From the time of embryo freezing until export, the in vitro produced embryos in this consignment were stored for at least 30 days:
    i) in sealed sterile containers (e.g. straws, ampoules or vials) and identified in a legible and non-erasable manner as specified in the current IETS Manual. Goblets and canes were also identified as specified in the current IETS Manual
    ii) Either:
    only with other bovine germplasm collected for export to Australia, or of equivalent health status
    OR
    with other export certifiable germplasm provided ALL straws, ampoules or vials were sealed and intact.
    iii) in storage or shipping containers containing only new, unused liquid nitrogen
    iv) in a secure place within a CFIA approved facility or laboratory
    v) under the supervision of the Team Veterinarian
    *20.2. For transport of embryos for further processing (e.g., DNA testing) at another facility, the in vitro produced embryos were:
    i) stored in sealed sterile containers (e.g. incubators, straws, ampoules or vials) and identified in a legible and non-erasable manner as specified in the current IETS Manual, and
    ii) processed hygienically under veterinary supervision
    (*Delete clause if embryos not transported for further processing; manual deletion must be initialled by the Official Government Veterinarian.)
21. Shipping containers
    Reproductive material was placed for final shipment in either:
    *21.1. a new shipping container,
    or
    *21.2. immediately prior to loading, the shipping container was emptied and inspected and any loose straws removed. The shipping container, including all surfaces in contact with the straws, ampules or vials was then disinfected with one of the following disinfectants:
    i) 2% available chlorine (e.g. chlorine bleach);
    ii) 2% Virkon;
    iii) 2.4% Prevail;
    iv) irradiated at 50kGy.
    Date of disinfection/ irradiation (yyyy-mm-dd):
    
    ……………………………….……………………………….……………………………….
    Disinfectant used/active ingredient:
    
    ……………………………….……………………………….……………………………….
    (*Delete unused option; manual deletion must be initialled by the Official Government Veterinarian.)
22. Official Government Seals
    22.1. Under the supervision of an Official Government Veterinarian prior to export to Australia:
    i) the containers (e.g. straws, ampoules or vials) for reproductive material in this consignment were checked as being sealed;
    ii) the identity of the reproductive material was checked prior to being placed into new, unused liquid nitrogen in a shipping container for export that was new or disinfected as specified in this veterinary certificate;
    22.2. Only bovine reproductive material (that is, semen and/or in vivo derived embryos and/or in vitro-produced embryos) that meet Australian import conditions can be transported in the same shipping container;
    22.3. An official government seal was applied by an Official Government Veterinarian to the shipping container and the number or mark on the seal recorded on this certificate.
    Shipping container official government seal number……………………………….