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Test selection and sampling requirements for aquatic animal imports

Information for foreign authorities

The purpose of these tables is to advise foreign authorities which diagnostic tests must be used when exporting susceptible aquatic species to Canada where testing or surveillance is a requirement and indicated on the approved zoosanitary export certificate.

These test selection and sampling requirements apply to all end uses that require testing for export or surveillance with the exception of food service, retail use, and further processing for human consumption.

Trade partners must select either the World Organisation for Animal Health (WOAH; founded as Office International des Épizooties (OIE)) or Canadian test indicated below. For diseases not listed by the WOAH, countries are expected to use the identified "Canadian test" below. Test selection or sample size may only be changed in situations where Canada has performed a country evaluation or assessed a country's aquatic animal health regulatory controls as part of the negotiated requirements for export to Canada.

All sampling must be completed using a screening test on apparently healthy populations. Tissues and sample size requirements are also indicated in the table below.

Pooling is not allowed; except for oyster seed and post-larval shrimp when conducting PCR testing on individual animals is not possible.

Please note

The WOAH tests listed below are based on the WOAH Manual of Diagnostic Tests for Aquatic Animals disease chapters sections 5 "Rating of tests against purpose of use". Generally, screening tests with an a) or b) in the "Targeted surveillance" column of the WOAH manual are the recommended tests for import.

Sample size has been determined based on either 1 or 2% prevalence and 95% confidence and takes into account disease clustering.

This document is subject to change based on new scientific evidence and development of more advanced protocols.

Finfish diseases

Table 1: The test type, sample size and sample type required by disease for the import of susceptible species of finfish into Canada
Diseases WOAH test Canadian test Sample size CFIA types of tissue samples, and/or lifestage
Infectious salmon anaemia virus (ISAv) pathogenic strains and non-pathogenic strains (HPR0)

Screening test:
Real time RT-PCR
(Snow et al method)

Confirmatory test: cRT-PCR (segment 6) and sequencing and virus isolation (HPR deleted strains)

Screening test:
qRT-PCR Table Note 1 (segment 8)

Confirmatory test: cRT-PCR (segment 6) and sequencing and virus isolation (for HPR deleted strains)

Originating from a Country or zone that is free, to all zones Table Note 2 in Canada: 175 for pathogenic ISA and 350 for HPR0

Originating from a Country or zone that is not free or has an unknown health status to a zone Table Note 2 in Canada that is free, provisionally free or a buffer zone:
Refuse entry Table Note 3

Originating from a Country or zone that is not free or has an unknown health status to a zone Table Note 2 in Canada that is infected:
175 for HPR deleted ISA and 350 for HPR0 ISA

Lethal testing-Pathogenic strains: Mid- kidney and heart HPR0: gill Table Note 4
Nonlethal broodstock testing:
Pathogenic strains: ovarian fluid, milt, HPR0: gill Table Note 4
Viral haemorrhagic septicemia virus

Screening test:
Virus isolation

Confirmatory test: PCR with sequencing to determine strain

Screening test:
qRT-PCR

Confirmatory test: cRT-PCR and sequencing and virus isolation

175 Lethal testing -Anterior kidney and brain; spleen and heart can be added if larger sample required for virus isolation
Non-lethal broodstock testing: ovarian fluid, milt
Infectious haematopoietic necrosis virus

Screening test: Virus isolation

Confirmatory test: PCR

Screening test:
qRT-PCR

Confirmatory test: cRT-PCR and sequencing and virus isolation

175 Lethal testing -Anterior kidney and brain; spleen and heart can be added if larger sample required for virus isolation
Non-lethal broodstock testing-eggs, ovarian fluid, milt
Infectious pancreatic necrosis virus Table Note 5 N/A

Screening test:
qPCR

Confirmatory test: cPCR and sequencing and virus isolation

Originating from a Country or zone that is free to all zones Table Note 2 in Canada:
175

Originating from a Country or zone that is not free or has an unknown health status to a zone Table Note 2 in Canada that is free, provisionally free or a buffer zone:
350

Originating from a Country or zone that is not free or has an unknown health status to a zone Table Note 2 in Canada that is infected:
175

Lethal testing - spleen and kidney
Non-lethal broodstock testing- ovarian fluid, milt Table Note 6
Epizootic haematopoietic necrosis virus

Screening test: Virus isolation

Confirmatory test: PCR

Screening test:
qPCR

Confirmatory test: cPCR and sequencing and virus isolation

175 Lethal testing – anterior-mid kidney and spleen
Oncorhynchus masou virus disease (Oncorhynchus masou virus)

Screening test: Virus isolation

Confirmatory test: PCR confirmation

As per the WOAH Manual of Diagnostic Tests for Aquatic Animals

Screening test:
qPCR

Confirmatory test: cPCR and sequencing and virus isolation

175 Lethal testing -anterior – mid kidney, skin lesions and spleen
Non-lethal broodstock testing- ovarian fluid
Gyrodactylosis (Gyrodactylus salaris)

Screening test: Visual inspection and wet mount

Confirmatory test: PCR and sequencing

Screening test:
cPCR and morphometry

Confirmatory test: cPCR and sequencing

175 Whole fish and/or fins
Koi herpes virus

Screening test:
PCR

Confirmatory test: PCR and sequencing

Screening test:
qPCR

Confirmatory test: cPCR (that targets a different segment of the genome) and sequencing

175 Anterior kidney, spleen, brain, gill and intestine
Spring viraemia of carp virus Screening test: Virus isolation

Screening test:
qRT-PCR

Confirmatory test: cPCR and sequencing and virus isolation

175 Kidney spleen, gill and brain
Red sea bream iridovirus Screening test:
PCR with sequencing confirmation

Screening test:
qPCR

Confirmatory test: cPCR and sequencing and virus isolation

175 Anterior - mid Kidney and spleen
Ceratomyxosis (Ceratomyxa shasta) Table Note 5 N/A

Screening test:
Visualization of parasite on gills or in gut smear

Confirmatory test: PCR

175 Gills, pancreas, pyloric cecae and adjacent intestine
Whirling disease (Myxobolus cerebralis) Table Note 5 N/A

Screening Test:
Pepsin Trypsin digest

Confirmatory test: cPCR and sequencing and visualization of spores

175 Head including gill arches and operculum
Salmonid alphavirus

Screening test:
Real time PCR

Confirmatory test: cPCR sequencing confirmation

Screening test:
qPCR

Confirmatory test: cPCR and sequencing and serology

175 Heart and kidney

Table Notes

Table Note 1

Canada requires testing for ISAV HPR0 using gill as the sample tissue and using the method described in Snow et al. 2006 New Diagnostic Technology: Applications in Animal Health and Biologics Controls Dev Biol 126:133-135.

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Table Note 2

Finfish Reportable Diseases-Declarations by Province and Marine Area health status in Canada

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Table Note 3

While diagnostic test methods are available for ISA HPR0, due to the transient expression and seasonal nature of this strain, and the difficulty in detecting infection when the prevalence is low, it is not possible at this time through testing alone to reduce the importation risk of ISA HPR0 to negligible.

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Table Note 4

Gill must be run independently of other tissues sampled.

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Table Note 5

This disease is listed in the Canadian Reportable Disease Regulations

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Table Note 6

Smail DA1, Munro ES. Isolation and quantification of infectious pancreatic necrosis virus from ovarian and seminal fluids of Atlantic salmon, Salmo salar L. J Fish Dis. 2008 Jan;31(1):49-58.

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Mollusc diseases

For diseases of molluscs that are considered as foreign animal diseases in Canada, histology alone does not have sufficient sensitivity for screening of apparently healthy populations.

Table 2: The test type, sample size and sample type required by disease for the import of susceptible species of molluscs into Canada.
Disease WOAH test Canadian test Sample size CFIA types of tissue samples, and/or lifestage
Abalone viral mortality (abalone herpes-like virus)

Screening test:
PCR

Confirmatory test:
in situ hybridization or PCR and sequencing

Screening test:
qPCR

Confirmatory test:
cPCR and sequencing and In situ hybridization

175 Adult animals: Mouth and the neural tissue of the cerebral commissure, pleuropedal and buccal ganglia (juveniles and adults).
Larvae, spat use whole animals (qPCR only).
Bonamia exitiosa

Screening test:
PCR

Confirmatory test: PCR with sequencing for confirmation

Screening test:
qPCR

Confirmatory test: cPCR and sequencing and histology

175 Adult animals: 3-5mm section through gill and mantle, gonad and digestive gland.
Larvae, spat use whole animals.
Bonamia ostreae

Screening test:
real-time PCR

Confirmatory test: PCR with sequencing for confirmation

Screening test:
qPCR or histology

Confirmatory test: cPCR and sequencing and histology

175 Adult animals: Gill, mantle, gonad and heart for PCR; Sagittal section through gill, digestive gland for histology.
pools of 2-3 ml for spat.
Haplosporidium nelsoni Table Note 7 N/A

Screening test:
qPCR or histology

Confirmatory test: cPCR and sequencing and histology

Originating from a Country or zone that is free, to all zones Table Note 8 in Canada: All susceptible species: 175 except
C. gigas: 350

Originating from a Country or zone that is not free or has an unknown health status to a zone Table Note 8 in Canada that is free, provisionally free or a buffer zone:
Refuse entry

Originating from a Country or zone that is not free or has an unknown health status to a zone Table Note 8 in Canada that is infected:
175 all species except C. gigas: 350

Adult animals: Gill, mantle, digestive gland for PCR; Sagittal section through gill and mantle, heart and digestive gland for histology.
2-3 ml pools for spat.
Marteilia refringens

Screening test:
PCR

Confirmatory test: PCR and sequencing or in situ hybridization

Screening test:
qPCR

Confirmatory test: cPCR and sequencing and histology

175 Adult animals: 3-5mm section through gill, mantle, gonad and digestive gland.
Larvae, spat use whole animals.
Marteiliodes chungmuensis Table Note 9 N/A

Screening test:
Histology

Confirmatory test: cPCR and sequencing

175 Adult animals ( female only): 3 to 5mm section through gills, mantle, ovarian tissue, and digestive gland
Mikrocytos mackini Table Note 7 N/A

Screening test:
qPCR or histology

Confirmatory test: cPCR and sequencing and histology

175 Adult animals: Sagittal section through base of gills, digestive gland, gonad, mantle and gills for PCR and histology.
Ostreid herpesvirus 1

Screening test:
qPCR

Confirmatory test: cPCR and sequencing (WOAH Manual of Diagnostic Tests for Aquatic Animals)

Screening test:
qPCR

Confirmatory test: cPCR and sequencing and In situ hybridization

175 Adult animals: 3 to 5mm section through gill, mantle, digestive gland and adductor muscle.
Larvae, spat use whole animals.
Perkinsus marinus

Screening test:
PCR

Confirmatory test:
In situ hybridization
or PCR with sequencing

Screening test:
qPCR

Confirmatory test: cPCR and sequencing and histology

175 Adult animals: 3-5mm section through gill, mantle, gonad and digestive gland.
Perkinsus olseni

Screening test:
PCR

Confirmatory test:
In situ hybridization
or PCR with sequencing

Screening test:
qPCR

Confirmatory test: cPCR and sequencing and histology

175 Adult animals: 3-5mm section through gill mantle, gonad and digestive gland.
Withering syndrome of abalone (Xenohaliotis californiensis)

Screening test:
PCR

Confirmatory test: PCR and sequencing

Screening test:
PCR

Confirmatory test: cPCR and sequencing and In situ hybridization

175 Adult animals: Posterior oesophagus and digestive gland.

Table Notes

Table Note 7

This disease is listed in the Canadian Reportable Disease Regulations

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Table Note 8

Mollusc Reportable Diseases-Declarations by Province and Marine Area

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Table Note 9

This disease is listed in the Canadian Health of Animals Regulations Schedule VII

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Crustacean Diseases

Histopathology alone cannot be used for confirmation of screening test in apparently healthy populations because histopathology does not have sufficient specificity.

Table 3: The test type, sample size and sample type required by disease for the import of susceptible species of crustaceans into Canada.
Diseases WOAH test Canadian test Table Note 10 Sample size CFIA types of Tissue samples, and/or lifestage
Taura syndrome virus

Screening test:
RT-qPCR

Confirmatory test: In situ DNA probes hybridization

Screening test:
qPCR

Confirmatory test: cPCR and sequencing and In situ hybridization

175 Whole post-larvae or
antennal gland, lymphoid organ, cuticular epithelium of gills, exoskeleton, appendages and anterior foregut for lethal sampling;
hemolymph, gills and pleopod for non-lethal sampling
White spot syndrome virus

Screening test:
RT-qPCR

Confirmatory test: In situ hybridization

Screening test:
qPCR Table Note 11

Confirmatory test: cPCR and sequencing and In situ hybridization

175 Whole post-larvae or
antennal gland, hematopoietic organ, cuticular epithelium of gills, exoskeleton, appendages and anterior foregut for lethal sampling;
hemolymph, gills and pleopod for non-lethal sampling
Yellowhead virus

Screening test:
RT-qPCR

Confirmatory test: In situ hybridization

Screening test:
qPCR

Confirmatory test: cPCR and sequencing and
In situ hybridization

175 Whole post-larvae or
lymphoid organ and cuticular epithelium of gills, exoskeleton, appendages and anterior foregut for lethal sampling;
hemolymph, pleopods and gills for non-lethal sampling
Crayfish plague (Aphanomyces astaci)

Screening test:
RT-qPCR

Confirmatory test: cPCR and sequencing

Screening test:
qPCR Table Note 11

Confirmatory test: cPCR and sequencing and histology (microscopic examination)

175 Soft abdominal cuticle, uropod and telson
Infectious hypodermal and hematopoietic necrosis virus

Screening test:
RT-qPCR

Confirmatory test: In situ hybridization

Screening test:
qPCR

Confirmatory test: cPCR and sequencing and In situ hybridization

350 Whole post-larvae or
antennal gland, hematopoietic organ, lymphoid organ, cuticular epithelium of gills, exoskeleton, appendages and anterior foregut for lethal sampling;
hemolymph, gills and pleopod for non-lethal sampling
Infectious myonecrosis virus

Screening test:
RT-qPCR

Confirmatory test: In situ hybridization

Screening test:
qPCR

Confirmatory test: cPCR and sequencing and In situ hybridization

175

Whole post-larvae or
white muscle, and lymphoid organ for lethal sampling;
hemolymph and pleopod for non-lethal sampling

White tail disease

Screening test:
RT-qPCR

Confirmatory test: In situ hybridization

Screening test:
qPCR

Confirmatory test: cPCR and sequencing and In situ hybridization

175 Whole post-larvae or striated muscle from cephalothorax, abdomen and tail. for lethal sampling;
pleopod for non-lethal sampling
Necrotizing hepatopancreatitis

Screening test:
RT-qPCR

Confirmatory test: In situ hybridization

Screening test:
cPCR Table Note 11

Confirmatory test: cPCR and sequencing and In situ hybridization

175 Whole post-larvae or hepatopancreas
Acute hepatopancreatic necrosis disease (AHPND)

Screening test:
real time PCR (Preliminary enrichment culture for detection of VpAHPND)

Confirmatory test: histopathology or bioassay

Screening test:
qPCR Table Note 11

Confirmatory test: cPCR and sequencing and In situ hybridization

175 Whole post-larvae or hepatopancreas, stomach

Table Notes

Table Note 10

Molecular assays are described in: Arseneau, J.R. and Laflamme, M. 2016. Development of RT-qPCR methodologies for the detection of viral pathogens in crustaceans of commercial importance. Can. Tech. Rep. Fish. Aquat. Sci. 3134: iii+21pp.

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Table Note 11

Method is published in the WOAH Manual of Diagnostic Tests for Aquatic Animals.

Return to table note 11  referrer

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