Importer Guidance for Aquatic Animal Health Import Quarantine Unit

1 Introduction

The purpose of this document is to provide importers with guidance when establishing a quarantine unit for approval by the Canadian Food Inspection Agency (CFIA) for international imports.

Key elements for quarantine unit creation include the following:

if possible, a hydrogeological survey of groundwater supplies;
a site diagram indicating the location of the physical infrastructure on the premises, the water flow patterns for the site and the locations of the different populations of animals on the site, if multiple populations are present;
written standard of procedures and written records that demonstrate compliance with the operating procedures; and
written records that demonstrate the implementation of active observational surveillance and traceability.

The importer should have quarantine unit staff that are trained and knowledgeable in the areas of biosecurity and aquatic animal diseases, or have a working relationship with an aquatic animal health professional or veterinarian.

This document is not meant to be a comprehensive list of requirements and is to be used as guidance for the establishment of a quarantine unit. The specific requirements of a particular quarantine unit are dependent on the individual premises, location of the quarantine unit, and life stage/species/product being imported.

1.1 Definitions

100-year flood: is the peak or flood flow with one chance in one hundred of occurring in any given year. (Crue centenaire)
Access points: initial control points with defined entry and exit procedures. (Points d'accès)
Active Observational Surveillance: the process of actively and systematically looking for diseased animals by a trained individual on a frequent, pre-planned, and ongoing basis, where a predefined plan of action is implemented when affected animals are discovered. (Surveillance par observation active)
Ante-room: is a predefined area that separates the quarantine unit from the surrounding environment. It may not be a room, but rather a designated area on the site where exterior clothing is exchanged for site specific clothing and a hand wash station is provided. (Antichambre)
Aquatic animal: means any finfish, mollusc or crustacean, or any part of a finfish, mollusc or crustacean at any life stage, as well as any germplasm of those animals. (Health of Animals, Part XVI, Section 190) (Animal aquatique)
Biological vector: is an organism, usually an arthropod, in whose body the pathogen develops or multiplies before becoming infective for the recipient individual. (Vecteur biologique)
Competent Authority: means the Veterinary Authority or other Governmental Authority of a Member having the responsibility and competence for ensuring or supervising the implementation of aquatic animal health and welfare measures, international health certification and other standards and recommendations in the Aquatic Code in the whole territory. (Aquatic Animal Health Code 2012)(Autorité compétente)
Effluent water: is the waste water from an aquatic animal tank or holding system. Refer to "Solid waste." (Effluents)
Equipment: refers to all inanimate objects that can be used in the quarantine unit, including but not limited to equipment for aquatic animal handling, feeding, mortality collection and disposal as well as person protective equipment such as boots and raingear, graders, and fish pumps. (Équipement)
Fomite: Any inanimate object or substance capable of carrying infectious organisms. This may include but is not limited to equipment, vehicles and articles of clothing or shoes. (Vecteur passif)
Germplasm: means semen, male or female germ cells or genetic material taken from a male or female germ cell for the purpose of producing a zygote and includes embryos but does not include a hatching egg. (Health of Animals Regulations) (Matériel génétique)
Import: means entry into Canada from any other origin. (Importation)
Influent water: is the inflow of water into an aquatic animal tank or holding system. (Eau d'arrivée)
Inorganic waste: is material that is non-decomposable and does not consist of animal and/or plant matter. (Déchets inorganiques)
Inspection: means an inspection carried out by an inspector, except where a customs officer carries out an inspection pursuant to section 16 of the Health of Animals Act. (Health of Animals Regulations) (Inspection)
Inspector: means a person who is designated as an inspector, pursuant to subsection 13(3) of the Canadian Food Inspection Agency Act, for the purposes of section 32 of the Health of Animals Act and/or subsection 17 of the Fish Inspection Act. (Inspecteur)
Mechanical vector: is a living organism that can passively transmit the pathogen to an aquatic animal but is not infected by the pathogen (and therefore cannot actively shed the pathogen). (Vecteur mécanique)
Organic waste: is material that is decomposable and consists of animal and/or plant matter. (Déchets organiques)
Pathogens of concern: refers to the infectious agents in which the aquatic animals are considered to be susceptible. (Agents pathogènes préoccupants)
Permit: refers to an aquatic animal health import permit issued pursuant to Section 160 of the Health of Animals Act. (Permis)
Population: means a group of aquatic animals that are linked by direct or indirect contact with each other. The number of populations in a quarantine unit is determined by the links between the animals within the quarantine unit and the risk factors (animals, water, feed, fomites and vectors) that influence the health status of the quarantine unit. (Population)
Reportable: means prescribed as reportable by the minister as listed in the Reportable Diseases Regulations. (À déclaration obligatoire)
Risk assessment: means the evaluation of the likelihood and the biological and economic consequences of entry, establishment and spread of a hazard within the territory of an importing country. (Aquatic Animal Health Code 2013) (Évaluation des risques)
Sample: means a defined number of aquatic animals taken from a population of aquatic animals for the purpose of testing or collection of specimens. (Échantillon)
Solid waste: is inorganic or organic material that is present in transportation, influent, and effluent water. (Déchets solides)
Specimen: means a tissue, organ or bodily fluid removed from an aquatic animal for the purpose of testing. (Spécimen)
Veterinary inspector: means a veterinarian designated as an inspector pursuant to section 32 of the Health of Animals Act. (Vétérinaire-inspecteur)

2 Creation of a quarantine site diagram

Quarantine site diagrams are required for all quarantine units unless otherwise stated by the CFIA. Site diagrams can be hand-drawn or computer-generated. It is recommended that a base diagram of the physical infrastructure is first created upon which the other elements can be superimposed (e.g. water flow, animal population, and traffic flow patterns) as it is recognized that these elements may change. Refer to Appendix A: Quarantine Site Diagram for an example diagram of each of the following three steps listed below.

Add basic physical infrastructure
Main components in a quarantine site diagram:
- Quarantine unit or building encompassing the quarantine unit
- Separate rooms in the quarantine unit (e.g. office, laboratory, and feed storage)
- Water supply
- Infrastructure (e.g. location of aeration towers, water treatment, shipping and receiving areas, and parking)
- Aquatic animal holding area
- Effluent water (e.g. settling ponds, septic tanks, and discharge location)
- Water flow pattern
- Restrictions of movement of people, animals, or vehicle traffic (e.g. gates and fences
Superimpose water flow pattern
Add aquatic animal locations

3 Creation of standard operating procedures

Appendix C: Standard operating procedure guidelines for import quarantine consists of a list of standard operating procedures (SOP) and their guidelines for a quarantine unit. The importer is to identify SOPs applicable to their particular quarantine unit from the list in Appendix C and additional procedures and more information may be required under certain circumstances as determined by the CFIA. Not all of the SOPs listed necessarily apply to every quarantine unit, and in some cases SOPs can be combined.

3.1 How to write a SOP

A SOP provides a short set of instructions sufficient to tell staff members how to conduct the procedure. It is primarily an instructional document. Detailed reference material to support the SOP can be appended if desired, but is not necessary; the document's primary function is to instruct staff in simple terms how to carry out the required procedure.

A SOP contains several usual elements that tell the user the following:

who is responsible for conducting the procedure;
what to do;
how to do it;
required equipment or supplies and where they are located;
frequency to conduct the procedure; and
the critical parameters/occurrences that require immediate action or notification of management.

Refer to Appendix B: Example Standard Operating Procedure (SOP) for Quarantine for an example SOP.

3.1.1 SOP elements

The following headings may be included in each written procedure, although not all are necessary every time. Importers may wish to add additional sections or information for their own purposes.

Title:: Provide a descriptive title.
Rationale:: Explain why the procedure is being conducted.
Definitions:: List any definitions or acronyms required to understand the SOP.
Responsibility:: Define who is responsible for the information in the SOP and who is responsible for conducting it. Use position titles or people's names to indicate responsibility.
Equipment and supplies:: List all equipment required (if any) to complete the SOP.
Safety concerns:: List any worker safety or animal health concerns that must be addressed.
Procedure:: Describe, in active voice, each step required to complete the SOP, the frequency with which it is conducted, and any critical findings/occurrences that require notification of management or implementation of a contingency plan.
Contingency plan:: Outline the actions to be taken should a critical finding occur (e.g. an influent screen found to be damaged must be replaced or repaired as soon as possible).
Record keeping:: List all records required, where and how they are stored, and how long they must be maintained. Provide an example of how to fill out records, if desired.

3.1.1.1 Record-keeping requirements

Records are kept to demonstrate that the required quarantine unit procedures are conducted. Record keeping is variable for each activity.

3.2 Guidance applicable to the creation of a quarantine unit

3.2.1 Animals

3.2.1.1 Unintentional Animal Introductions into the Quarantine Unit

Influent and effluent screens are necessary to prevent inadvertent animal introductions; however, there are circumstances where they may not be required (e.g. influent screens are not required if water source is municipal water or ground water; and effluent screens are not required if water is treated and there is a physical barrier preventing invasion of all aquatic animals into the quarantine unit through the effluent system). Screen size is recommended to be no larger than the minimum size of aquatic animal that is possibly present in the influent and effluent water source. Choice of screen size will be a consideration when quarantine units request the acceptance of screening as a physical barrier to prevent invasion of other aquatic animals.

Influent and effluent screens must undergo routine inspections and maintenance and/or repairs as necessary to prevent the unintentional introduction of aquatic animals into the quarantine unit. Influent and effluent screens must be cleaned and maintained on a regular basis to ensure their effectiveness. The screens must be clear of debris and must not exhibit damage.

3.2.1.2 Intentional Animal Introductions into the Quarantine Unit

All aquatic animal introductions must be in compliance with import permit conditions and only the aquatic animals listed on the import permit can be introduced into the quarantine unit. A single quarantine unit must contain only the aquatic animals to be imported as specified on the import permit and a single quarantine unit can contain more than one population with the written preapproval of the CFIA. If multiple populations are introduced into a quarantine unit during the quarantine period, they are considered to have the same health status. The quarantine unit must demonstrate traceability so that in the event of a disease outbreak, trace-in and trace-out can be completed.

3.2.1.3 Aquatic Animal Gametes and/or Germplasm Treatment

Aquatic animal gametes and/or germplasm must be treated prior to export in the foreign country or upon arrival into the quarantine unit. The products used for treatment must be effective against the pathogens of concern specific to the species being imported, compliant with international standards, used in accordance with manufacturer's recommendations, and approved by the CFIA.

Importers are strongly encouraged to follow the World Organisation for Animal Health (WOAH; founded as Office International des Épizooties (OIE)) recommendations for surface disinfection of salmonid eggs in the WOAH Aquatic Animal Health Code, 24th Edition, 2021, Chapter 4.4.

3.2.2 Water

3.2.2.1 Unintentional Introduction of Water into the Quarantine Unit

The quarantine unit should be located in an area that is outside the area to be affected by a 100-year flood. If the quarantine unit resides within a 100-year flood area, sufficient dikes/levees must surround it to prevent flooding or other mitigation measures must be implemented.

A 100-year flood is the peak or flood flow with one chance in one hundred of occurring in any given year. Refer to Environment Canada and provincial governments for identification of flood areas and risks applicable to the region of the quarantine unit.

3.2.2.2 Treatment and Disposal of Water and/or Ice Accompanying Imported Aquatic Animals for Quarantine

Whenever possible, transportation water should be disinfected within transportation containers after the transfer of aquatic animals into the quarantine unit. If applicable, transportation ice must be melted prior to treatment. Low levels of suspended solid waste present in the transportation containers can be disinfected and disposed of with the transportation water. Significant amount of solid waste present in the transportation containers must be settled or filtered out of the water in order for the water treatment to be efficacious. Solid waste must either be stored in a leak-proof holding container within the quarantine unit until the aquatic animals are released from quarantine or treated with an approved method to destroy the pathogens of concern prior to disposal.

The method selected to treat transportation water must have supporting evidence, such as scientific literature, demonstrating it as efficacious against pathogens of concern. The transportation water can be introduced into the quarantine unit if the effluent treatment system in the quarantine unit can sufficiently handle the volume of transportation water and effectively treat the pathogens of concern. In order to determine if the effluent treatment system can handle the volume of transportation water, the importer must assess the effluent treatment system in the quarantine unit and provide evidence of its capacity and capabilities.

3.2.2.3 Assessment of Influent Water Source

Water sources that are pathogen free and acceptable as influent water sources for a quarantine unit include ground water, spring water, de-chlorinated municipal water supplies, and marine premises that use manufactured sea salts (such as Instant Ocean) mixed with pathogen free freshwater. Other sources of water such as surface water would need to be screened, filtered, and treated by a method demonstrated to be effective for elimination of pathogens of concern to be acceptable for use.

All influent water sources for the quarantine unit would be subject to an assessment prior to approval of the quarantine unit. For ground or spring water sources, information needed for the assessment could include records pertaining to the well depth (if applicable), land use for surrounding area (and local industries), water source classification (confined aquifer or unconfined aquifer), the history of the water quality, the changes in water temperature over the course of one year and other water-quality parameters, the soil or rock type into which the well is drilled, dug or bored, and any data available for the testing of the water for the presence of pathogens of concern.

The likelihood of groundwater being contaminated by surface water supplies depends on the soil/rock types in which the wells have been dug, drilled or bored; the depth of the well; and the distance from surface water supplies. The make-up of the confining bed over the ground water supply influences the ability of surface water to penetrate the substrate and reach ground water. Where the confining bed is impermeable, creating a confined aquifer, it is improbable that contaminated surface water has reached the water in the confined aquifer. Where the aquifer is unconfined, that is the confining bed is permeable, it is more likely that the ground water supply can be contaminated by surface waters. Water temperature changes indicate the proximity of the water to the surface and deep wells will show virtually no temperature fluctuation. Shallower wells will have greater temperature fluctuation (up to 5°C to 10°C).

Reference:

Heath, RC. Basic Ground Water Hydrology. US Geological Service Water Supply Paper 1983:2220.

3.2.2.4 Water Quality Monitoring

The maintenance of water quality is critical to the quarantine unit and specific water quality parameters can impact the water treatment method chosen and the efficacy of water treatment, equipment disinfection, and egg disinfection procedures. At a minimum, water temperature, pH, and turbidity must be monitored and maintained within a range that is acceptable according to the treatment system in the quarantine unit. Water quality monitoring, including the parameters to be measured and the frequency of measurement, is specific to the quarantine unit and will vary with the water source and type of system (e.g. recirculation versus flow through).

Water quality should be monitored consistently and accurately, in order to be confident in the water quality parameters that impact water treatment, egg disinfection and equipment disinfection activities. Acceptable parameters vary depending on the disinfection methods used for water treatment, egg disinfection or equipment disinfection. The importer should take into account the natural, seasonal water quality changes. Water quality should be measured frequently enough to differentiate normal variation from declining water quality conditions. Acceptable water quality parameters and the expected actions of staff if they encounter an unacceptable result should be predetermined.

Water quality parameters that may impact the efficacy of influent, effluent and transportation water treatment are as follows:

pH;
water temperature;
turbidity;
total dissolved solids;
total suspended solids;
iron manganese;
hydrogen sulphide;
calcium magnesium;
residual ozone levels; and
any other parameters that need to be measured to ensure disinfection systems are effective.

3.2.2.5 Influent and Effluent Water Treatment

Influent and effluent water treatment must ensure log 4 reduction of the pathogens of concern.

Acceptable influent water sources include municipal water that is treated to drinking water standards and ground water that is proven to not be influenced by surface water. Other sources of influent water are acceptable when pre-filtered and treated using one of the following methods that is effective to destroy the pathogens of concern to which the aquatic animals are susceptible: chlorination/de-chlorination, ultraviolet light, ozone, autoclave, electrolytic water treatment, and any other method that is demonstrated to be sufficient to destroy the pathogens of concern and is subsequently approved by the CFIA.

Effluent water must be pre-filtered and treated using one of the following methods that is effective to destroy the pathogens of concern to which the aquatic animals are susceptible: chlorination/de-chlorination, ultraviolet light, ozone, autoclave, electrolytic water treatment, and any other method that is demonstrated to be sufficient to destroy the pathogens of concern and is subsequently approved by the CFIA.

Settling and filtration

Influent water sources other than municipal and ground water must be pre-filtered prior to treatment. Effluent water must always be pre-filtered prior to treatment.

The selection of settling and filtration systems is dependent on the characteristics (type, size, and density) of the particles suspended in the water supply and the flow rate of the system.

Efficacy of water settling and filtration prior to disinfection methods are determined via:

evaluation of manufacturer's specifications;
documentation of routine required maintenance;
monitoring of turbidity or organic loading of the systems prior to the chosen water treatment method; and
specific pre-treatment requirements of each disinfection system.

Chlorination/de-chlorination

The level of organic matter and nitrogenous wastes in the water affect the amount of chlorine that must be added to the system; these compounds affect the ability of the chlorine molecules to disinfect. Organic matter and nitrogenous wastes must be measured and removed from the water prior to chlorination.
Monitoring of the free chlorine levels in the system is done at least daily, both for efficacy and for prevention of toxicity to aquatic animals in the rearing system.

Ultraviolet disinfection

Sufficient sizing of the UV system: there must be sufficient UV C output performance (typically requiring low pressure bulbs), including accounting for 40% loss of bulb strength over the bulb lifespan. UV C is a subtype of Ultraviolet light.
Transmissibility of the water to be disinfected must be known (90-95% transmittance required); therefore, pre-filtration of most water supplies is required.
The operating water flow rate must be known in order to calculate the actual dosage being delivered.
Dosage and exposure level, thus the sizing of the system, must be targeted to the pathogen(s) of concern.
The frequency of UV bulb changes must follow manufacturer's specifications.
Quartz sleeve maintenance requirements depend on the water quality, thus the probability of scale and biofilm deposits on the sleeve. Maintenance schedules must follow manufacturer's recommendations.
All water to be treated must pass through a filter capable of removing suspended organic material prior to irradiation; this can include use of sand, drum, bead, or other physical filters, reverse osmosis and/or ozone treatment of the water to increase particle size for ease of removal.
Specifications of the individual systems with measurement of the water transmittance is used as the determination of whether a system is acceptable for the specific situation.

Disinfection using Ozonation

The ozone demand of the treatment system is dependent on flow rate and water quality. The ozone demands must be known by the quarantine unit staff as pre-filtration and ozone concentration/exposure is varied depending on the system demands.
Ozone levels must be monitored to verify successful levels achieved.
Residual ozone must be removed prior to contact with aquatic animal populations.

Electrolysis

Currently, electrolysis is not used extensively.
NaCl must be present in the system.
Measurement of residual hypochlorite and hypobromite is required to determine ongoing efficacy.

Other methods

Other methods can be considered and require approval by the CFIA.

The importer must demonstrate proper treatment system maintenance for the chosen quarantine unit treatment method.

3.2.2.6 Disposal of Solid Waste Collected from the Quarantine Unit

Solid waste is inorganic or organic material that is present in transportation, influent, and effluent water.

Solid waste must be stored in secure containers within the quarantine unit until it is treated to destroy the pathogens of concern. Waste may be disposed of during the quarantine period upon approval by CFIA inspection when treated prior to disposal; the treatment system must be sufficient to destroy the pathogens of concern to which the aquatic animals are susceptible for the quarantine unit. Several treatment systems are acceptable for quarantine, including rendering, incineration, sterilization, or another method approved by the CFIA. The quarantine unit must either have disposal to municipal sewer or septic system after treatment or disposal to landfill after treatment. The quarantine unit must comply with all provincial and municipal waste management regulations.

3.2.3 Feed

3.2.3.1 Feed Types and Sources

The following feed types are acceptable:

Live and/or raw unprocessed feed sources from species not susceptible to the pathogens of concern for the species that the feed is being fed to;
Live and/or unprocessed feed sources that have been certified by the competent authority at the origin of the feed source as free for the pathogens of concern for the species that the feed is being fed to; and
Dried, pelleted feed sources that have undergone a manufacturing process that is sufficient to destroy the pathogens of concern for the species that the feed is being fed to. Dry, commercially produced feed where total moisture content is less than 10% are negligible risk feed sources.

All feed must be from sources or manufacturers that have methods in place to protect against cross contamination with other feeds, aquatic animals or aquatic animal products between source and delivery at the quarantine unit. Any aquatic animal feed source that is not one of the acceptable feed sources listed above being used in the quarantine unit must undergo a risk assessment by the CFIA prior to its approval for use.

Dry Pelleted, Extruded Feed and Flake Feeds

In general:

Dry pelleted and extruded feeds generally represent a negligible risk of pathogen introduction due to the temperatures reached during the manufacturing processes for the production of fish meal (75°C to 100°C) and the extrusion/pelletizing process itself; temperatures reached during extrusion can range from 90°C to >120°C which is sufficient to kill all pathogens of concern.
Flake feeds are highly processed, cooked and dried, and thus represent a negligible risk of pathogen introduction.

Moist or Semi-Moist Feeds

Due to the moisture content of moist and semi-moist feeds and the lower temperatures of processing, the risk of pathogen transfer in these feeds is elevated over that of dry or extruded pellets.
The manufacturing process for moist feeds is typically a cold process, and thus the aquatic animal carcasses/offal used for manufacturing must come from fish sources that are determined by the CFIA to represent a negligible risk of pathogen introduction if these feeds are used.

Live Feeds

If live aquatic animals are to be used as feed for aquatic animals within a quarantine unit, the live feed must be of negligible risk for the pathogens of concern OR if considered susceptible to the same diseases of concern as the aquatic animals in the quarantine unit, the live feed must be certified as free of diseases of concern prior to use as feed.
Once live animals to be used as aquatic animal feed are introduced into a quarantine unit, they are considered to have the same health status as the aquatic animal population in the quarantine unit.

Offal/Aquatic Animal Waste/Lyophilization

Raw offal or aquatic animal waste used as aquatic animal feed in the quarantine unit requires risk assessment by the CFIA to determine the associated risk and required level of testing of the feed source. In general this is not an acceptable feed source as the likelihood of pathogen introduction is significant.
Raw Offal/fish waste used as feed should not consist of aquatic animal species that are susceptible to any pathogens of concern to which the aquatic animals in quarantine are susceptible.
Aquatic animal waste/offal should be sourced from negligible risk sources and testing of the waste/offal must occur for the pathogens to which the source, recipient, and local aquatic animal populations are susceptible.
Freeze dried feeds – lyophilization preserves many pathogenic micro-organisms and thus requires the same risk assessment as offal or aquatic animal waste. For example, infective IPNv may be preserved for many years in freeze dried form depending on the storage medium (Wolf et al 1969).

Cross-Contamination Prevention

Feeds must be protected from cross-contamination with other feeds or aquatic animal products prior to delivery at the quarantine unit in order to prevent pathogen introduction.
Feeds must be obtained from a reputable source and stored in a manner that protects the feed from extremes of heat, light, and humidity at all times from manufacturing to feeding.

References:

Whipple and Rohovec. The effect of heat and low pH on selected viral and bacterial fish pathogens (report that IPNv will be killed with a process that reaches 65°C for 15 minutes followed by 5 minutes at 82°C). Aquaculture 1994;123:179-189.

Nygaard, H, Myrmel, N, Myrmel, M. Inactivation of fish pathogenic micro-organisms in fish by-products. Sub-Project IPN-virus. Nofima Report no. 30/2010. 2010; ISBN: 978-82-7251-801-0.

Wolf, K, Quimby, MC, Carlson, CP. Infectious pancreatic necrosis virus: lyophilization and subsequent stability at 4°C. Appl. Micr. 1969;17(4):623-624.

3.2.3.2 Feed Storage

Proper storage and handling of aquatic animal feed is essential to decrease predator and scavenger attraction to the premises. Feed to be used for multiple populations must be managed to prevent cross-contamination with feeds that are to be used in the quarantine unit.

Transportation vehicles should be cleaned and disinfected prior to picking up new feed supplies if the same vehicle is used for animal handling (live or dead).

Feed should be stored with a secure lid. Spilled feed, including excess oil leakage from feed bags, should be cleaned up immediately to decrease scavenger attraction to the quarantine unit. The feed storage area should be accessed daily in order to look for signs of access by scavengers/pests; if there are visible signs of scavengers/pests, preventive measures should be implemented such as checking that access points are blocked off, setting rodent traps, or calling the contract exterminator.

3.2.4 Fomites

3.2.4.1 Vehicle Management

Vehicles that contact aquatic animals, aquatic animal waste, equipment, or staff may become contaminated with pathogens of concern. Vehicle access to the quarantine unit must be limited to essential vehicles and vehicle parking areas must be clearly identified. Those vehicles that transport aquatic animals and/or enter the quarantine unit are subject to cleaning and disinfection procedures.

Water, detergent, and scrub brushes should be used to remove all visible organic matter from the outside surfaces of the vehicle prior to disinfection to ensure efficacy. Wheel wells and tires should not be missed. The cleaned vehicle should be rinsed with pathogen free water and allowed to dry before application of the disinfectant. The disinfectant should be applied and left in place for a duration as indicated on the manufacturer's instructions. The disinfectant should then be rinsed off with pathogen free water and left to dry.

Managing the quarantine unit equipment within the vehicle can remove the need for cleaning of the entire vehicle interior. Specific areas of the vehicle should be designated for quarantine unit materials to prevent cross contamination. Areas of the vehicle which are easily disinfected (e.g. trunk or box) should be used for transport of quarantine unit materials and all quarantine unit equipment should be placed into the designated quarantine unit area of the vehicle (e.g. in totes, in plastic bags etc.). Only the areas of the vehicle that have been in contact with the quarantine unit equipment and/or materials require cleaning and disinfection.

Disinfectants should be disposed of according to manufacturer's directions and meet the requirements of local waste management regulations.

3.2.4.2 Equipment Management

Equipment is to be kept clean and disinfected to limit the spread of pathogens of concern. Separate equipment must be designated for a single quarantine unit. All new equipment introduced to the quarantine unit must be cleaned and disinfected before use. Equipment should be designated for the quarantine unit and labelled with colour coding or name of the location. Cleaning and disinfection protocols must be followed for equipment that is transferred into and out of the quarantine unit. Acceptable disinfection methods for a quarantine unit include fallowing, drying, ultraviolet light, chemical disinfection or any other method approved by the CFIA. Products should be used according to the manufacturer's directions. Organic matter should be removed from equipment prior to disinfection to ensure efficacy. Disinfected equipment should be rinsed with pathogen-free water and allowed to dry.

Disinfectant concentrations should be maintained by checking concentration (test strips) or regular renewal of the product. Disinfectants should be disposed of according to manufacturer's directions and the requirements of waste management regulations.

3.2.4.3 Inorganic Waste Management – Disposal

Inorganic waste is material that is non-decomposable and does not consist of animal and/or plant matter.

Inorganic waste must be stored in secure containers and can be released from the quarantine unit without treatment once the aquatic animals are deemed to be of negligible risk for the pathogens of concern after the completion of disease freedom testing. Inorganic waste presents less risk of disease transfer out of the quarantine unit than organic waste. Inorganic waste disposal must be conducted in a manner that prevents the spread of the pathogens of concern outside of the quarantine unit. Several options exist for acceptable waste disposal including landfill (properly covered), incineration, and any other method that contains the waste from the aquatic animals on site and the surrounding environment. The quarantine unit must follow provincial and municipal requirements for waste disposal. Waste may be disposed of during the quarantine period upon approval by CFIA inspection if treated prior to disposal; the treatment system must be sufficient to destroy the pathogens of concern to which the aquatic animals are susceptible for the quarantine unit.

3.2.4.4 Cleaning and Disinfection of Equipment and Materials of the Quarantine Unit

The quarantine unit, animal holding units, and equipment must be constructed of materials that can be disinfected to ensure destruction of the pathogens of concern. Aquatic animal holding units and their plumbing located inside the quarantine unit must be constructed from materials that can be cleaned and disinfected. Preferred materials include plastics, fibreglass, and metals. Lower preference is porous materials (e.g. concrete).

Transportation materials and packaging must be made of materials that can be disinfected or made of disposable materials to ensure destruction of pathogens of concern. Aquatic animals and their products must be held in containers that are made from materials that can be cleaned and disinfected prior to entry and exit of the quarantine unit. Preferred materials include plastics, fibreglass, and metals. Lower preference is porous materials (e.g. concrete).

All equipment, packaging materials, vehicles or any other items which come in contact with the animals or water from the quarantine unit must be cleaned and disinfected in a manner which prevents the spread of pathogens of concern out of the quarantine unit. Several disinfection methods are acceptable including chemical disinfectants, physical disinfectants, or other methods approved by the CFIA that have been demonstrated as efficacious for destruction of the pathogens of concern. Chemical disinfectants must be labelled as effective for the destruction of the pathogens of concern and used according to manufacturer's directions. Physical disinfection procedures must reach the time and temperature profiles sufficient to destroy the pathogens of concern (e.g. steam cleaning). The entire quarantine unit must be cleaned and disinfected in a matter that is sufficient to destroy the pathogens of concern before the arrival of the aquatic animal population into the quarantine unit.

3.2.4.5 Chemical Storage

Chemical disinfectants must be stored in a manner that prevents their degradation and maintains efficacy. Manufacturer's instructions must be followed for proper storage of chemicals. The chemicals should be stored in a dry, secure area and in a location with the manufacturer's recommended room temperature. Expiry dates of chemicals should be noted before use and those expired should be properly discarded.

3.2.5 Vectors

3.2.5.1 Access Control

People, equipment or objects must not have access to the quarantine unit without the knowledge and control of the importer. Access to the quarantine unit must be prevented using a fence, locked door or another method that demonstrates that access is controlled. Signage must be visibly posted to indicate restricted access into the quarantine unit; the signage must be clear, legible and provide directions to the defined entry area or contact information for the office. The signage must be within the quarantine unit property boundary.

Anyone other than quarantine staff must contact the importer prior to entering the quarantine unit.

3.2.5.2 Quarantine Unit Biosecurity

Entry and exit procedures must be developed and implemented to prevent the introduction and spread of pathogens of concern on hands, feet, and clothing of all persons entering and exiting the quarantine unit. An ante-room is required as part of the entry and exit procedures. The goal of the ante-room is to provide a predefined transition area between the quarantine unit and the surrounding environment. The ante-room is not required to be a separate room from the quarantine unit and can be a designated area on the site or a clearly marked and segregated area within the quarantine unit. The ante-room area must be cleaned and disinfected regularly (at pre-defined intervals or upon accumulation of any organic matter). Site-specific clothing and footwear or the use of footbaths in entry and exit procedures of the quarantine unit is required and must be located in the ante-room. Disinfection stations must be provided and utilized in the ante-room and at all entry and exit points of the quarantine unit. Footbaths must be properly maintained, assessed daily for signs of fouling with organic material, and changed at regular intervals because they can become contaminated with micro-organisms and become a source of pathogen introduction. Only clean boots/shoes should be dipped in the footbath as footwear covered in organic matter will not be successfully disinfected. Where available, testing strips should be used daily to assess the footbath disinfectant concentration. Manufacturer's directions should be followed for the dilution of the disinfectant and staff provided with the container volumes and volumes of powdered concentrate or liquid disinfectant solution to add to a known volume of water (e.g. for a 1% solution, add 10 grams of powder per litre of water in the footbath).

It is preferred to have staff members work solely at the quarantine unit; however, if they must travel between the quarantine unit and other premises or rooms outside the quarantine unit, procedures must be implemented to prevent transfer of pathogens of concern on those staff members and their outerwear. Those wishing to enter the quarantine unit must be informed that they cannot have had contact with live aquatic animals or aquatic animals products, including wild aquatic animals, outside of the quarantine unit or been on another aquatic animal premises for 48 hours prior to visiting the quarantine unit.

A quarantine access logbook must be filled out by any persons wishing to enter the quarantine unit, which includes the attendance of previous aquatic animal locations within the past 48 hours. Quarantine staff must accompany anyone else who enters the quarantine unit at all times while on site and ensure they complete required the entry and exit procedures. Quarantine staff should inform those that enter the quarantine unit that they are not to handle aquatic animals, feed, and equipment. Access to sensitive areas (feed and mortality storage) should be restricted to quarantine staff unless access is required for inspection purposes. Refer to the example of a quarantine access logbook in Appendix C.

3.2.5.3 Predator, Scavenger, and Pest Control

Preventive measures and procedures must be in place to prevent access of predators, scavengers, and pests into the quarantine unit. The quarantine unit should use predator exclusion devices tailored to the specific needs of the site. Visible portions of the holding units should be checked, including the predator exclusion devices daily. Any damage to holding units or predator exclusion devices should be repaired as soon as possible. Feed should be stored and distributed in a manner that does not increase attraction to scavengers and predators and spilled feed should be cleaned up immediately. Waste must be properly stored prior to removal from the site and visible mortalities must be collected daily and stored securely and held separately from the aquatic animals in order to prevent access to predators, scavengers, and pests.

3.2.5.4 Mortality Management – Collection and Storage

The presence of mortalities can result in attraction of predators and scavengers and have negative effects on water quality and sanitation in the environment. Routine removal of mortalities minimizes pathogen spread. All visible dead and moribund aquatic animals must be removed on a daily basis and must be maintained in the quarantine unit in a biosecure location until disease freedom testing is completed and/or quarantine is released by CFIA Inspector. The dead animals must be identified, counted, and separated by container and/or unit. The containers used to collect and store mortalities must have secure, tight fitting lids that will exclude predators and scavengers, be leak-proof, and should be stored in an area away from the live aquatic animals. Moribund animals (animals exhibiting clinical signs or in a dying state) must be humanely euthanized prior to testing and/or disposal. Hygienic precautions should be taken when handling dead aquatic animals, which include wearing gloves while handling dead aquatic animals and washing hands after mortality removals.

The quarantine unit must have its own equipment that is associated with mortality collection, storage, and disposal and must be appropriately cleaned and disinfected according to manufacturer's instructions.

3.2.5.5 Mortality and/or Carcass Management – Disposal

Mortality and/or carcass disposal must be conducted in a sanitary manner to prevent the spread of the pathogens of concern to aquatic animals into or out of the quarantine unit. Various disposal methods are acceptable, including landfill (only if the animals are tested in quarantine and determined to be free of pathogens of concern), composting (only if the animals are tested in quarantine and determined to be free of pathogens of concern), autoclaving, ensiling, incineration, and any other method upon approval by the CFIA.

3.2.6 Specific Procedures for Active Observational Surveillance of the Quarantine Unit

Aquatic animals must be monitored and visually inspected daily by a trained individual who actively and systematically looks for diseased animals on a frequent daily basis while the animals are in quarantine. Moribund animals or mortalities must be collected daily and visually inspected by trained personnel for diseases of concern. Individual mortalities should be routinely examined internally for abnormalities or clinical signs of disease. The CFIA Inspector should be contacted when observations of clinical signs of disease occur. CFIA disease cards for specifics of diseases to which the quarantined species is considered susceptible should be available for staff

Procedures for active observational surveillance for aquatic animals:

daily monitoring of behaviour (including feeding activity) and morbidity;
daily mortality collection with specific instructions for when staff should contact management;
routine, gross internal examination of mortalities; and
emergency plan.
Visually monitor for the following aquatic animal behaviours (if applicable):
feeding;
schooling;
reaction to human presence;
flashing;
gasping; and
visible, gross clinical signs of disease.

Mortalities should be classified and can include the following:

systems-related (e.g. water quality);
predator;
handling/transport;
clinical signs of disease—describe signs;
fresh—freshly dead with no clinical signs of disease; and
old—no longer suitable for diagnostic testing.

Specific triggers for the quarantine unit that indicate a serious disease occurrence:

clinical signs of septicaemia (haemorrhages);
ascites;
trailing fecal casts;
gray-coloured gills in live fish;
presence of granulomas or other tissue lesions; and
any other specific triggers for an individual site.

Tissue specimen collection for laboratory testing must be performed by CFIA inspectors.

Quarantine staff should be trained and knowledgeable of the biosecurity procedures and aquatic animal diseases to which the species are susceptible to prevent the introduction and release of pathogens of concern to and from the quarantine unit. The staff should be knowledgeable and experienced in monitoring aquatic animal health and the recognition of abnormal aquatic animal behaviour or clinical signs of disease. The quarantine unit should have an identified aquatic animal health professional or veterinarian associated with the quarantine unit who can be consulted during the quarantine period.

Appendix A: Example quarantine site diagram

Example: 1. Add basic infrastructure. Description follows. — **Example: 1. Add basic infrastructure**

Example: 2. Superimpose water flow pattern. Description follows. — **Example: 2. Superimpose water flow pattern**

Example: 3. Add animal locations. Description follows. — **Example: 3. Add animal locations**

Any locations that restrict the movement of people, animals or vehicle traffic can be further superimposed upon the diagram (e.g. gates, fence lines, etc.).

Example: 4. Laboratory Quarantine Room. Description follows. — **Example: 4. Laboratory Quarantine Room**

Appendix B: Example standard operating procedure for quarantine

This example of a generic procedure must be modified by the importer to the specifics of the quarantine unit, and their implementation will be verified during quarantine unit inspections.

This SOP is a generic example of the information that needs to be covered in a biosecurity plan. Each item that is italicized prompts the importer to enter site-specific information for the premises.

Water quality monitoring

Note: The maintenance of water quality is critical to good fish health management. However, this written procedure is limited to those water quality parameters that may impact the efficacy of water treatment and the disinfection of newly fertilized or eyed eggs and equipment. This information can be combined with the water treatment or egg disinfection SOPs, if desired.

Rationale: The efficacy of water treatment, equipment disinfection, and egg disinfection procedures are impacted by specific water quality parameters.

Definitions: List any definitions or acronyms required to understand the SOP.

Responsibility: Identify the staff names or positions that are responsible for the information in the SOP and for carrying out the procedure.

Required equipment: List all required equipment and locations; e.g. pH test strips, pH meter, refractometer, sample bottles, laboratory submission forms, reagents, glasswear, etc.

Procedure: The goal of this SOP is to ensure that water quality is monitored consistently and accurately, in order to be confident in the water quality parameters that impact water treatment, egg disinfection and equipment disinfection activities.

Acceptable parameters vary depending on the disinfection methods used for water treatment, egg disinfection, and equipment disinfection. Add the specific parameters that are commonly measured on your premises; e.g. turbidity prior to UV disinfection, pH prior to egg disinfection.

The water quality monitoring program should be designed to consider natural, seasonal water quality changes and the premises' specific needs. Describe the parameters measured routinely or those that change throughout the year due to changing conditions.

Water quality should be measured frequently enough to differentiate normal variation from declining water quality conditions. Clearly identify what level of each parameter is acceptable and the expected actions of staff if they encounter an unacceptable result.

pH is measured prior to egg disinfection and if chlorination/de-chlorination is used as a water treatment method.

Measure pH (insert frequency) using pH test strips/pH meter.

Describe calibration method for pH meter, or provide a web link or include a copy of the manufacturer's instructions.

Identify where in the system to collect the water sample.

Provide instructions for recirculation systems where the water quality is continually monitored on how to monitor the system, how to verify that chemical stores automatically added by the system are sufficient, and how to conduct calibrations.

Adjust pH to neutral using (insert chemicals or methods used to modify pH; e.g. sodium bicarbonate/calcium carbonate or flushing with fresh water to decrease un-ionized ammonia concentrations).

Provide the amount of chemical to add to a given volume of water, or calculations required for the specific chemical added.

Water quality parameters that may impact the efficacy of water disinfection are as follows:

total dissolved solids;
total suspended solids;
iron manganese;
hydrogen sulphide;
calcium magnesium;
residual ozone levels; and
add any other parameters that need to be measured to ensure disinfection systems are effective.

Describe procedures used to monitor each of the above water quality parameters if they are a concern for your site, including frequency of testing. These parameters are measured after water filtration but before the water enters the UV unit. Identify the location in the water treatment system to collect a sample.

Water quality testing on site is conducted as follows:

calibrate and maintain equipment (provide staff with manual); and
identify levels of each water quality parameter that require action on the part of staff; e.g. TDS <800 to 1000 g/l; TSS <10 mg/l; Iron <0.3 mg/l; Manganese <0.05 mg/l; H2S—non-detectable; Ca/Mg <120 mg/l CaCO₃.

Safety concerns: Provide staff with MSDS sheets for all chemicals used in water quality monitoring. Identify safety issues around use of UV (e.g. voltage) and ozone disinfection systems (risks of human exposure to ozone).

Contingency plan: Identify actions that staff is required to take in the event of deteriorating water quality.

Record-keeping requirements:

identify records kept; e.g. fish health, results of diagnostic testing; and
identify where records are stored.

Appendix C: Standard operating procedure guidelines for import quarantine

Note: The SOPs that are not applicable to dead aquatic animal carcasses or parts thereof are denoted with an asterisk (*).

This is table is not intended to be a complete list of requirements for a particular quarantine unit. Additional SOPs may be required after the CFIA has conducted the documentation and physical inspections.

Standard Operating Procedures

Animals
Import Quarantine Procedures	Details
Unintentional animal introductions into the quarantine unit*	SOP: screen maintenance and repair; and proper screen size. Record-keeping: Date of maintenance and repair(s).
Intentional animal introductions into the quarantine unit	Record-keeping: Date and source of aquatic animal introduction into the quarantine unit – specific origin, name of supplier/country; If sampling and testing for aquatic animal health purposes is required, any and all records related to the sampling and testing, including diagnostic results must be kept. Date and destination of animal movements into and out of the quarantine unit. All records must be kept for two years of import; All movements of aquatic animal population(s) within the quarantine unit; Inventory and list of species in each aquatic animal holding unit; and Original import permit.
Aquatic animal gametes and/or germplasm treatment*	SOP: treatment procedure; when to implement the treatment; the reason for the treatment; identification of the staff member or position responsible for carrying out the procedure; an adequate description of the required dilutions to reach the therapeutic concentrations of disinfectant/treatment solutions, including maintenance of acceptable water quality of the solution and maintenance of solution concentrations; an adequate description of any hazards for treatment (e.g. high levels of organic matter or high pH that interfere with the efficacy of egg disinfection solutions); disinfectant/treatment solution is maintained as per manufacturer's recommendations; effluent disposal (if applicable); and water-quality parameters specific to the species, life stage and treatment being carried out. Record-keeping: Any information where the application of the SOP varies from the accepted protocol; Reason for the treatment; Exact name of the product used; Names and/or trade names of the products used, including dosages/concentrations; and Written records documenting the date(s) of treatment and the following information: life stage of the animals; location of treated animal groups; and specific water-quality parameters that may be relevant to the treatment type. Maintain records for a minimum of two years

Water
Import Quarantine Procedures	Details
Unintentional introduction of water	SOP: if applicable, mitigation measures to prevent flooding.
Treatment and disposal of water and/or ice accompanying imported aquatic animals for quarantine	SOP: the method, location and equipment used for treatment and disposal of transportation water and/or ice and solid waste (if present); Record-keeping: Any deviations from the SOP must be recorded; If water is disinfected within the transportation container, the method of water disinfection, including the dosages achieved (chemical) and exposure time must be recorded; If transportation water and/or ice is introduced into the quarantine unit, the flow rate of the treatment system and volume of transportation water introduced into the treatment system must be recorded to support that the volume of transportation water introduced into the quarantine unit treatment system did not affect its efficacy. Calculations to support the above must be performed and confirmed prior to the import and the introduction of the transport water and/or ice; Evidence for acceptance of disposal of transportation water into the quarantine unit; and The method and volume of water treated.
Assessment of influent water source*	Record-keeping: If the water source for the quarantine unit is ground water or spring water, the following information should be recorded in order to determine whether the source is uncontaminated: the use of a well; the depth of the well; land use for the surrounding area, and local industries that may affect inputs; the water source classification (confined aquifer or unconfined aquifer); the history of the water quality; the annual changes in water temperature and other water-quality parameters; the soil or rock type into which the well is drilled, dug or bored; and any data available for the testing of the water for the presence of pathogens (e.g. E. coli).
Water quality monitoring	SOP: the parameters that are to be measured; the methods and equipment used; the frequency of monitoring required for the location; and the normal expected water quality parameters, including upper and lower limits, and required actions in the event of an unacceptable change in water quality. Contingency plan: Deterioration in specific water quality parameters are identified and specific measures must be implemented to correct the problems (e.g. if the transmittance of water decreases below 90% in a UV system, the drum filter may need cleaning or sand filter may need backwashing); Water flow interruption (overflow, leakage); and Changes in water quality that indicate the contamination of a ground water supply or failure of a water treatment system require the quarantine unit to contact CFIA. Record-keeping: Type and frequency of water quality measurements; Calibration of water quality monitoring equipment when required; and Actions taken to amend water quality problems.
Influent and effluent water treatment*	SOP: details on pre-filtration, including the type of filter and size of particles removed as well as the method and frequency of cleaning of the filter; and for water treatment, the specific parameters routinely monitored for the system, and any maintenance required. Note: The specifics that should be measured and the frequency of measurement are dependent on the system used (e.g. when UV disinfection is used, transmittance should be monitored and follow manufacturer's guidelines for frequency). Contingency plan: Outlines each component of the system that is prone to failure, for example planning for UV bulb replacement and power outages. Reporting to CFIA if there is water treatment system failure. Record-keeping: Records must be kept for all maintenance, failures or repairs to the water treatment system.
Disposal of solid waste collected from the quarantine unit	SOP: information on the waste collection equipment and staff outerwear, including cleaning and disinfection procedures; handling and holding of the waste collected, waste disposal location, equipment used for holding waste prior to treatment, and treatment methods used prior to disposal of the waste from the quarantine unit; service companies provided with the quarantine unit biosecurity guidelines to follow, such as instructions for where to pick up waste or drop off supplies; and how to provide the company information on where the conveyance has been prior to coming to the site; biosecurity guidelines for service companies, including instructions for where to pick up waste or drop off supplies; and how to provide the quarantine unit information on where the conveyance has been prior to coming to the site; contact information for the waste collection service if one is used; and the waste disposal location, equipment and measures taken to ensure waste is covered when disposed of. Record-keeping: Date of waste pickups or disposal on-site; Contact information for driver or vehicle; and Biomass of waste collected (estimate or weight if available).

Feed
Import Quarantine Procedures	Details
Feed storage*	SOP: live feed procedures (if applicable); how feed is protected from scavengers and vermin, including vermin control program implementation when necessary; methods for disinfection of feed containers; handling spilled feed and clean-up and disposal; and methods for prevention of cross contamination of feed sources (e.g. separate containers with lids). Record-keeping: Records indicate when signs of scavengers and vermin are seen and the control methods implemented; and Invoices for feed for the quarantine unit, including the feed lot number, date of manufacture and delivery date.

Fomites
Import Quarantine Procedures	Details
Vehicle management	SOP: the identification of parking locations for vehicles and signage procedures for entrance and exit into the quarantine unit; cleaning and disinfection procedures for vehicles that enter and exit the quarantine unit, including cleaning and disinfection location, methods, types of surfaces on which disinfection will be completed, and products used; vehicle disinfection location; requirement to maintain on-site vehicles/vessels in clean condition to minimize the transfer of pathogens on people and equipment travelling in the vehicle/vessel; the use of impermeable transfer boxes for equipment when possible so that they can be effectively disinfected; the choice of travel routes and logistics to minimize pathogen transfer; and indication of the location for vehicles to conduct pick-up or drop-offs, and wherever possible, have this done at a distance from the quarantine unit and separate "dirty" from "clean" activities. Record-keeping: A logbook of the vehicle movements is required; Notation in the logbook of vehicle disinfection activities is required; and The attendance of service sector vehicles to the quarantine unit (e.g. mortality pickups or feed delivery must be noted in the quarantine access log).
Equipment management	SOP: quarantine unit-specific equipment identification and inventory maintenance (e.g. quarantine equipment is marked with a large "Q" in indelible ink); and cleaning and disinfection of the quarantine unit equipment. Record-keeping: Maintain records of equipment: equipment used; location of transfer from origin to destination; cleaning and disinfection protocol used; and date of transfer and reason for the transfer.
Inorganic waste disposal	SOP: the waste disposal location, equipment and measures taken to ensure waste is covered when disposed of; information on the waste collection equipment and staff outerwear, including cleaning and disinfection procedures; if service companies pick up waste, the written procedure must outline the measures taken to minimize contamination on the site (e.g. pickup of waste containers at the quarantine unit perimeter and provision of a concrete pad that can be cleaned and disinfected after pickup); service companies are provided with the quarantine unit biosecurity guidelines to follow, such as instructions for where to pick up waste or drop off supplies, and how to provide the company information on where the conveyance has been prior to coming to the site; and contact information is available for the waste collection service if one is used. Record-keeping: Date of waste pickups or disposal on-site; Contact information for driver or vehicle; and Biomass of waste collected (estimate or weight if available).
Cleaning and disinfection	SOP: manufacturer's instructions for chemical handling, use of each type of chemical disinfectant (including contact time and frequency), and specific disinfectant procedures (e.g. cleaning prior to disinfection, contact time, rinsing disinfectant off equipment and drying after use) for of all equipment (for example: feeding, mortality, graders, and fish pumps), packaging materials, vehicles and other fomites that could possibly introduce pathogens of concern to aquatic animal populations outside the quarantine unit; process for advising and training staff on chemical handling and disinfection procedures; procedural requirements for heat treatment, which include the time and temperature parameters to be reached to destroy the pathogens of concern; and procedures for maintenance of disinfectant and frequency of renewal or exchange of disinfectant to sure its efficacy. Record-keeping: Disinfectants used; Receipts for disinfectant purchases; and Dates of equipment cleaning and disinfection.
Chemical storage	SOP: manufacturer's instructions for chemical disinfectant storage

Vector
Import Quarantine Procedures	Details
Access control	SOP: maintenance of the fences, doors, and signage; and procedure for checking that access points are secure at all times (e.g. doors are locked). Record-keeping: The dates of maintenance to fences, doors, and signage.
Quarantine unit biosecurity	SOP: Quarantine unit access procedures: requirement for contacting the quarantine unit prior to entry; providing copies of biosecurity requirements (including personal and cleaning and disinfection requirements) to anyone planning on entering the quarantine unit; asking those that intend on entering the quarantine unit if they have had any contact with aquatic animals or aquatic animal products within the previous 48 hours and the nature of the contact, and limiting quarantine unit access; Specific elements pertaining to staff clothing and raingear: requirements for personal clothing worn on site (e.g. is clean and dry); work clothes provided for staff (e.g. remains in the quarantine unit); and disinfection of personal raingear upon entry or exiting the quarantine unit, ensuring sufficient contact time with the chosen disinfectant. Specific elements pertaining to staff movements between quarantine units/other sites and logistics of daily activities: if staff are exposed to aquatic animals or their products outside of their work environment, precautions to prevent the movement of pathogens between systems (e.g. such as change of clothes, disinfection of raingear or a time restriction between activities); daily activity planning in a manner that decreases the risk of pathogen movement from the quarantine unit (e.g. the order of daily activities in the staff and site biosecurity procedures); and high risk activity planning (e.g. mortality handling and disposal are done by separate staff from those staff doing feeding if possible). ante-room procedures; disinfection station procedures, which include a footbath and hand wash station be provided and in some cases designated footwear or clothing be present upon entry to and exit from the quarantine unit; instructions for using CFIA approved disinfectants (according to manufacturer's directions), which includes the disinfectant solutions being changed frequently enough to ensure efficacy; and instructions for disinfectant footbath dilution (specific to the disinfectant used). Record-keeping: The dates of cleaning and disinfection of the ante-room; A footbath logbook is present to document the dates of footbath changes and the disinfectant type and concentrations used; and A quarantine access logbook is required to detail all access to the quarantine unit, including the names of those entering, date, reason for visit, where they come from, and time since visiting last aquatic animal location.
Predator, scavenger, and pest control	SOP: where applicable, preventive measures used to eliminate predator access to outdoor quarantine unit (e.g. bird nets, lidded tanks, electric fences); preventative measures used to eliminate scavengers and pests from accessing the quarantine unit; frequency of inspection and methods for evaluation of animal holding units and overall quarantine unit and repair of any damage (to prevent predator or scavenger access); instructions to follow mortality management SOP (e.g. remove mortalities daily, at a minimum, to decrease attractiveness of the site to scavengers and predators); waste storage; and instructions to follow spilled feed cleaning protocol. Record-keeping: Record access of predators and scavengers into the feed storage areas and other parts of the quarantine unit; Document preventive measures undertaken to prevent access; and Record damage to the quarantine unit and any related mortalities.
Mortality management – collection and storage*	SOP: instructions on hygienic precautions, including wearing gloves while handling dead aquatic animals, and washing and sanitizing hands after mortality collection; procedure and frequency for removal of dead aquatic animals from holding units; equipment use protocol; cleaning and disinfection of raingear and equipment used to remove mortalities in the quarantine unit (refer to cleaning and disinfection SOP); mortality collection, including counting and visually examining mortalities as they are collected; mortality storage; and mortality handling procedures.
Mortality and/or carcass management – disposal	SOP: Where approved by the CFIA, procedure for mortality disposal to a landfill or compost premises including the location, equipment used, and measures taken to ensure mortalities are covered when disposed of; cleaning and disinfection of mortality collection equipment and staff outerwear; if service companies pick up mortalities, there must be a procedure that outlines the measures taken to minimize contamination on the site (e.g. pickup of mortality containers at the quarantine unit perimeter, and provision of a concrete pad that can be cleaned and disinfected after pickup); quarantine unit biosecurity guidelines for service companies (e.g. instructions for where to pick up mortalities and waste or drop off supplies and how to provide the quarantine unit information on where the conveyance has been prior to coming to the site); and contact information for mortality collection service if one is used. Contingency plan: Mass mortality disposal. Record-keeping: Method for management and disposal of dead aquatics animals; Date of pickups for disposal or disposal if on-site; Contact information for driver or vehicle; and Biomass of mortalities collected (estimate or weight if available).

Active Observational Surveillance of the Quarantine Unit
Import Quarantine Procedures	Details
Daily aquatic animal monitoring*	SOP: daily monitoring of aquatic animal behaviour, including appearance and any signs of morbidity, including the following: feeding; schooling; reaction to human presence; flashing; gasping; visible gross clinical signs of disease; and any other specific signs the site staff normally observes daily. specific actions required on the part of the staff monitoring the aquatic animals, including informing the aquatic health manager or site manager. Record-keeping: Notation of abnormalities in behaviour or appearance; and Date and aquatic animal holding unit in which abnormalities were noted.
Visual examination of quarantine aquatic animal holding units for signs of disease*	SOP: protocol for daily collection, visual examination by trained personnel, and counting of moribund animals and mortalities; mortalities are counted, visually examined for clinical signs of disease, and their numbers are recorded daily; and protocol for when abnormal mortalities are observed (e.g. quarantine unit staff must report to the supervising Veterinary Inspector). Contingency plan: Mandatory notification of Veterinary Inspector when detect or suspicious of disease in the case of specifics signs of disease or an unexplained increase in morbidity or mortality rate. Record-keeping: Inventory records; Abnormalities noted through visual examination of moribund animals or mortalities; and Follow up measures for determining the cause of abnormal morbidity or mortality.
Routine internal visual inspection of individual aquatic animal mortalities*	SOP: procedure for when abnormalities are observed; where staff have specific training in aquatic animal health, procedure for the recording of pathological signs that are observed and assignment of specific triggers for the quarantine unit that indicate a serious disease occurrence. Some examples follow: clinical signs of septicaemia (haemorrhages); ascites; trailing fecal casts; gray-coloured gills in live fish; presence of granulomas or other tissue lesions; and any other specific triggers for an individual site. procedure for response when observations of clinical signs of serious disease occur. Record-keeping: Notation of all abnormalities or possible clinical signs of disease in daily records; and Diagnostic test results.
Staff training in biosecurity and aquatic animal diseases*	SOP: on-the-job training in the biosecurity plan for all staff; and experience or training program in recognizing the normal patterns of behaviour (feeding and schooling), productivity, and mortality so that staff recognize when something abnormal occurs.

Appendix D: Example quarantine access log

Date YYYY/MM/DD	Name and Phone Number	Company/Association	Last contact with aquatic animals or visit to another fish rearing facility	Time In	Time Out

Appendix E: Susceptible Species

The CFIA Susceptible Species of Aquatic Animals table provides a list of the scientific names of susceptible species and identifies the disease(s) to which they are susceptible.

Susceptibility in aquatic species is determined if the infection can be demonstrated by natural occurrence of the disease in the species, or by experimental exposure of the species to the disease agent through a pathway that mimics a natural route of infection.

It is required to immediately contact the CFIA if reportable diseases are detected or suspected in the aquatic animals of the quarantine unit.