Chapter 10 - Artificial insemination
10.2 Bovine
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This module describes the requirements to qualify donor for export eligible bovine semen, for domestic use and for owner's use only. On farm and isolation requirements are described when applicable.
Export eligible semen
On farm requirements for bovines entering a centre for the production of semen for export
The herd of origin must not be subject to any quarantine, and the animals must be subject to the following tests with negative results, unless otherwise specified, within a 60-day period before entry into the isolation facility of the centre:
- Tuberculosis – intradermal purified protein derivative (PPD) bovine tuberculin – caudal fold read at 72 hours
- any reaction must be reported to the Canadian Food Inspection Agency (CFIA) veterinarian
- Brucellosis – (serum) fluorescence polarization assay (FPA)) or test prescribed by World Organisation for Animal Health (WOAH; founded as Office International des Épizooties (OIE)) for international trade
Leptospirosis – (serum) microscopic agglutination test (MAT) – serotypes L. pomona, L. sejroe, L. hardjo, L. canicola, L. grippotyphosa and L. icterohaemorrhagiae
Bulls with a test result of 1:100 or 1:200 must be retested after a minimum interval of 14 days and a stable or declining titre obtained to be considered negative and eligible for entry into the isolation facility of the centre. The sampling dates and titres must be recorded on the health certificate that accompanies the animal to the Artificial Insemination (AI) centre.
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Bulls with a test result of 1:400 or higher must be retested at least twice at intervals of 14 days
Bulls that are negative at 1:400 on at least 2 consecutive retests are considered to have a stabilized low titre and can enter the isolation facility of the centre. Bulls may be tested multiple times in order to achieve the 2 consecutive low titre tests 14 days apart, provided the original tests stated above fall within the 60 day period prior to entry into the isolation center. The sampling dates and titres must be recorded on the health certificate that accompanies the animal to the isolation facility of the AI centre.
- Bluetongue – (serum) competitive enzyme-linked immunosorbent assay (c-ELISA)
- Johne's disease – (serum) ELISA
- Enzootic bovine leucosis (EBL) – (serum) ELISA
- Epizootic hemorrhagic disease of deer – Type 2 (EH02) – (serum) agar gel immunodiffusion (AGID)
Bovine viral diarrhea (BVD) – (serum) Immunoperoxidase (IP) and serum neutralization (SN). Animals under age 6 months must be retested by immunoperoxidase and SN after reaching that age
The SN test determines the serological status of the prospective donor bull. Seropositive and seronegative animals are allowed to enter into the isolation facility of the centre.
Optional test
Note: an Infectious Bovine Rhinotracheitis (IBR) test is mandatory for entering into an isolation facility of a bovine semen production centre that is approved for semen export to the European Community (EC).
- Infectious bovine rhinotracheitis (IBR) – (serum) ELISA
This test is performed at the request of the Artificial Insemination centre; a negative test is required for animals entering into an IBR negative facility.
EBL testing the dam
In addition, when possible, the uterine dam of the prospective donor bull should be subjected to an ELISA test for EBL, subsequent to the weaning of the prospective donor, with negative results.
This test of the uterine dam is required to export semen to the EC Member States when semen is collected from a donor bull before reaching age 24 months, and a negative result to an ELISA test is required after reaching that age. This test is not required when the prospective donor bull originates from a Canada Health Accredited Herd for EBL (CHAH–EBL).
Samples for EBL testing are to be sent to the CFIA Saint Hyacinthe Laboratory – Retrovirology Centre of Expertise. When sending samples from uterine dam(s) with samples from tested bulls, please specify in the "Comments and Special Instructions" box of the CFIA 5473 form the vials corresponding to the uterine dam(s) and instructions that they are to be tested for EBL only.
A CFIA/ACIA 1634 Certificate of Health for Entry into a Semen Production Centre must accompany the animal to the isolation facility of the centre. (Refer to 10.1 Artificial Insemination – General Information paragraph 11 for distribution).
Isolation health requirements for bovines entering a centre for the production of semen for export
All qualifying and testing procedures associated with the assessment of the isolation health status of semen donor bulls are to be performed by a veterinarian who is authorized to perform these functions as an accredited veterinarian. Before performing any inspection or test in an isolation facility, the accredited veterinarian must have reviewed this section with a Canadian Food Inspection Agency (CFIA) district veterinarian, and have been trained on site to collect and process samples for campylobacteriosis and trichomoniasis testing.
All animals presented for entry as additions to the resident herd of a semen production centre must undergo a minimum period of 30 days in an isolation facility maintained by the centre; the isolation period may begin when all animals are present in the isolation facility.
An animal is to be considered eligible for entry into isolation only if a certificate of health completed by an accredited veterinarian as stated in article 3, accompanies the animal to the isolation facility. An identification check is conducted on each animal against the certificate of health presented by the centre. The certificate of health must show that the animal has met the appropriate on farm requirements that correspond to the status of the centre he is expected to enter after the isolation procedure.
One blood sample is collected from each animal at the beginning of the isolation period for the IBR-ELISA test. The sample is immediately sent to the laboratory as no paired samples are required at the end of the isolation period.
The initial tests of preputial samples for campylobacter fetus subsp venerealis infection and trichomoniasis (Tritrichomonas foetus) are conducted after at least 7 days in isolation. Preputial fluid examinations should not be conducted until at least 7 days have elapsed following natural service or semen collection by means of an artificial vagina. Module 3.4 gives specific information on collection and processing of the sample.
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Campylobacter fetus subsp. venerealis examination requires the use of a special transport enrichment medium (TEM) and coordination with the laboratory when scheduling the submission of samples
Sample(s) of preputial washing are to be subject to culture test(s) as follows:
- animals less than 6 months of age kept since 30 days of age only in a single sex group prior to entry in isolation must be tested at least once with negative result
- all animals more than 6 months of age or those that could have been in contact with females after 30 days of age prior to entering isolation must be tested 3 times at weekly intervals with negative result
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Tritrichomonas foetus examination is by culture of preputial sample followed by direct microscopic examination
The culture examination requires the use of a special Tritrichomonas foetus transport medium (TFTM) and coordination with the laboratory when scheduling the submission of samples. Sample(s) of preputial washing are to be subject to culture test(s) as follows:
- animals less than 6 months of age kept since 30 days of age in a single sex group prior to entry in isolation must be tested at least once with negative result
- all animals more than 6 months of age or those that could have been in contact with females after 30 days of age prior to entering isolation must be tested at least 3 times at weekly intervals with negative result
While in the isolation facility and after a minimum of 30 days in isolation, the pre-entry tests must be repeated with negative results, unless otherwise specified, as follows:
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Tuberculosis – intradermal PPD bovine tuberculin – caudal fold read at 72 hour
A minimum of 60 days is required between the tuberculosis test done on-farm and this one. In the event of a suspicious or inconclusive tuberculin test during testing in the pre-entry isolation facility, a comparative cervical test must be performed by the CFIA. A caudal reaction confirmed to be negative to M. bovis by comparative cervical test performed by a CFIA veterinarian is considered negative for entry purposes.
- Brucellosis – (serum) FPA or test prescribed by the World Organisation for Animal Health (WOAH) for international trade
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Leptospirosis – (serum) – Microagglutination test (MAT), serotypes L. pomona, L. sejroe, L. hardjo, L. canicola, L. grippotyphosa and L. icterohaemorrhagiae
Bulls with a test result of 1:100 or 1:200 must be retested after a minimum interval of 14 days and obtain a stable or declining titre to be considered negative and eligible for entry into the centre.
Bulls with a test result of 1:400 or higher must be retested at least twice at intervals of 14 days
Bulls that are negative at 1:400 on at least 2 consecutive retests are considered to have a stabilized low titre and can enter the centre. Bulls may be tested multiple times in order to achieve the 2 consecutive low titre tests 14 days apart. The sampling dates and titres must be recorded on the health certificate that accompanies the animal to the AI centre.
- Bluetongue – (serum) c-ELISA
- Johne's disease – (serum) ELISA
- Enzootic bovine leucosis (EBL) – (serum) ELISA
EHD2 – (serum) – AGID
Supplementary test for bulls entering into a bovine semen production centre approved to export to the European Community: in addition to the AGID test, an additional SN test must be performed during isolation for bulls selected for entry in a centre approved for export to the European Community.
- BVD – (serum) – Immunoperoxidase (IP) and SN
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An agent identification test: a virus isolation test (immunoperoxidase) or another WOAH recommended or suitable test, with negative result
Only when all animals in the pre-entry isolation have obtained negative results for this test, may animals in the pre-entry isolation facility enter in the semen collection centre. Animals younger than 6 months of age at time of the test must be retested later after reaching that age.
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A serological test to determine the presence or absence of antibodies: serum neutralisation (SN) or another WOAH recommended or suitable test; test result may be either negative or positive
When no seroconversion occurs in animals which tested negative prior to entering pre-entry isolation facility, all animals (seronegative and seropositive) are allowed entry into the semen collection centre. When seroconversion occurs in an animal which tested negative prior to entering pre-entry isolation facility, all animals that remain seronegative are kept in pre-entry isolation facility until there is no more seroconversion in this group when another test is performed after a minimum of 21 days.
Seropositive animals may be allowed to enter the semen collection centre.
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For all samples to be analyzed in CFIA laboratories from bovine under isolation notification (IAI), an Excel spreadsheet will need to be completed and forwarded electronically in addition to the paper form CFIA/ACIA 5473. Please consult the "special procedure for submission to CFIA laboratories, applicable to samples from bovine tested in isolation" described in section 3.2.
Optional test
Note: this test is mandatory to enter in a bovine semen production centre approved for export of semen to the European Community.
- IBR – (serum) ELISA test: this test is done at the request of the semen production centre; a negative test is required for animals entering into an IBR negative facility.
In the event of a suspicious or positive reaction being reported during the pre-entry isolation testing regime, the individual animal is to be removed from the group. The remaining animals shall not be released for entry into the production centre until either the suspect animal has been confirmed negative or has been removed from the isolation and the negative status of the remaining animals reconfirmed by retesting them for the disease in question.
Bovine semen collected during the isolation period may be labelled for "Owner Use Only" and released immediately to the owner of the bull, if the centre or the collector has a permit to collect and process owner use semen. When collected during the isolation period, semen is eligible for distribution in Canada; this semen must be stored in a manner that precludes the semen from entering the export stream and must not be released until all campylobacteriosis and trichomoniasis tests results are known. The collection area in the isolation premises must be approved in advance by the district veterinarian for this purpose and documentation on file.
Removal of donors from isolation either to enter the resident herd following the tests referred to above or to leave the centre, must be done only with the permission of the district veterinarian. The centre must be in possession of documentation acknowledging the bull is eligible to leave isolation and confirming which facility status it may enter.
A bull resident in a semen production centre for distribution in Canada does not qualify to enter the isolation facility of an export semen production centre. Such a resident bull must comply with the pre-entry requirements for export eligible semen in order to enter the isolation facility.
Domestic use semen
On farm requirements for bovines entering a semen collection centre for the purpose of the production of semen for distribution in Canada
Herd of origin
The herd of origin must not be subject to any quarantine, and the animals must be subject to the following tests with negative results, unless otherwise specified, within the 60-day period preceding entry into the isolation facility of the centre. The animal must be tested in Canada for entry into the isolation facility.
Required tests
Tuberculosis – intradermal purified protein derivative (PPD) bovine tuberculin – caudal fold read at 72 hours
Any reaction must be reported to the CFIA veterinarian.
- Enzootic bovine leukosis (EBL) – (serum) ELISA
- Johne's disease – (serum) ELISA
- BVD – (serum) Immunoperoxidase. Animals under age 6 months must be retested after reaching that age.
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Bluetongue – c-ELISA
An ELISA-positive animal may be accepted in the isolation facility if 1 of the 2 conditions mentioned below apply:
- a negative result to a polymerase chain reaction (PCR) test is conducted on a blood sample
Note: this test is done at the expense of the owner in a private laboratory, and the result must be attached to the certificate mentioned in number 15 (below)
or - a longer period is applied to the animal in the isolation facility maintained by the centre; the animal must remain in isolation for a minimum of 60 days, and during this period, no semen from this animal can be collected, except for "Owner use only"
- a negative result to a polymerase chain reaction (PCR) test is conducted on a blood sample
A CFIA/ACIA 1634 certificate must accompany the animal to the isolation facility of the centre (Refer to paragraph 11 in Module 10.1 for distribution).
Isolation health requirements for bovines entering a centre for the production of semen for distribution in Canada
All qualifying and testing procedures associated with the assessment of the isolation health status of semen donor bulls are to be performed by a veterinarian who is authorized to perform these functions as an accredited veterinarian. Before performing any inspection or test in an isolation facility, the accredited veterinarian must have reviewed this section with a CFIA district veterinarian, and have been trained on site to collect and process samples for campylobacteriosis and trichomoniasis testing.
All animals presented for entry as additions to the resident herd of a semen production centre must undergo a minimum period of 30 days in an isolation facility maintained by the centre except for animals admitted with a positive c-ELISA result for Bluetongue without a negative PCR test, that must undergo a minimum isolation period of 60 days; the isolation period may begin when all animals are present in the isolation facility.
An animal is to be considered eligible for entry into isolation only if a certificate of health completed by an accredited veterinarian as stated in article 15 accompanies the animal to the isolation facility. An identification check is conducted on each animal against the certificate of health presented by the centre. The certificate of health must show that the animal has met the appropriate on farm requirements that correspond to the status of the centre he is expected to enter after the isolation procedure.
All animals presented for entry as additions to the semen production centre must receive an injection of dihydrostreptomycin (25 mg per kg of body weight) or an alternate antibiotic effective against leptospirosis at the dosage recommended by the manufacturer, as a preventive measure.
The initial tests of preputial samples for Campylobacter fetus subsp. venerealis infection and trichomoniasis (Tritrichomonas foetus) may be conducted when animals arrive in isolation. Preputial fluid examinations should not be conducted until at least 7 days have elapsed following natural service or semen collection by means of an artificial vagina. Module 3.4 gives specific information on collection and processing of the sample
Campylobacter fetus subsp. venerealis examination requires the use of a special medium (TEM) and coordination with the laboratory when scheduling the submission of samples.
Sample(s) of preputial washing are to be subject to culture test(s) as follows:
- animals less than 6 months of age kept since 30 days of age only in a single sex group prior to entry in isolation must be tested at least once with negative result
- all animals more than 6 months of age or those that could have been in contact with females after 30 days of age prior to entering isolation must be tested 3 times at weekly intervals with negative result
Tritrichomonas foetus examination is by culture of preputial sample followed by direct microscopic examination
The culture examination requires the use of a special transport medium (TFTM) and coordination with the laboratory when scheduling the submission of samples. Sample(s) of preputial washing are to be subject to culture test(s) as follows:
- animals less than 6 months of age kept since 30 days of age in a single sex group prior to entry in isolation must be tested at least once with negative result
- all animals more than 6 months of age or those that could have been in contact with females after 30 days of age prior to entering isolation must be tested at least 3 times at weekly intervals with negative result
Bovine semen collected from a bluetongue negative animal (defined as having had either a c-ELISA negative test or a c-ELISA positive test followed by a PCR negative test) during the isolation period may be either labelled "Owner Use Only" if the centre has the proper permit and released immediately to the owner of the bull or kept in the centre and released for distribution in Canada only when results of the all Campylobacter and trichomoniasis culture tests are negative and the minimum 30-day isolation period is over. Collection of semen from bluetongue positive animals (defined as having had a c-ELISA positive test with no subsequent negative PCR test or a subsequent positive PCR test) is not authorized before the mandatory 60-day isolation period is over. Any semen collection done during the isolation period must be authorized by the district veterinarian.
Removal of donors from isolation either to enter the resident herd following the tests and appropriate isolation periods referred to above or to leave the centre, must be done only with the permission of the district veterinarian.
A bull resident in a semen production centre for distribution in Canada does not qualify to enter the isolation facility of an export semen production centre. Such a resident bull must comply with the pre-entry requirements for export eligible semen in order to enter the isolation facility of an export eligible semen production centre.
Owner's use only semen
Requirements for bovines collected for owner's use only
Owner's use only semen is intended for use by owners of donor animals in their own herd. Owner's use bovine semen must only be collected and processed by a person or organization holding a Permit to collect and process animal semen for owner use only. This permit is issued by the National Coordinator of the Artificial Insemination program. There is no requirement to be an accredited veterinarian to obtain this permit. Further information may be obtained from the district office.
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