Security
People entering and leaving quarantine stations/confinement facilities may offer an efficient method for disseminating specified material. All personnel using a facility where specified materials or pests are being held should be suitably trained. They should be made aware of the work, by at least reading and signing an SOP, to ensure they do not inadvertently breach containment.
Access should be restricted to authorised personnel only. This is most easily achieved by having lockable outer doors; the use of keypads, electronic swipe cards, Yale type locks or padlocks should be encouraged. Mortice type locks are not appropriate, as they effectively provide a tunnel between the facility and the outside world.
Ideally specified material should not be left unattended. If specified material has to be left unattended clear signage and physical separation from non-specified material is required.
Signage should be installed and state that the specified material should not be moved and include the contact details of the responsible person.
Physical separation is another secure measure, and ideally the specified material should be locked away. If locking the material away is not possible then it should be placed in a suitably secure area, isolated from non-specified material and accompanied by clear signage e.g; in an incubator with a sign on the front, with the hood pulled down.
Hygiene
Protective clothing, such as laboratory coats, overalls, gloves and footwear must be worn as appropriate. These should be clearly marked and left within the facility on exit, preferably in a lobby area with hand-washing facilities and a footbath (where appropriate).
Used/contaminated clothing should be decontaminated prior to laundering, which could involve autoclaving (121°C [15psi] for 15 minutes) or in the case of invertebrates (other than those used to vector plant pathogens), freezing at -15°C for a minimum of 72 hours.
Disposable protective clothing is also acceptable but must be disposed of in line with the requirements listed on the scientific authorisation.
Pest control
There should be effective control of glasshouse pests which could disseminate specified material or pests, either directly as vectors or by the removal of contaminated material. This could be achieved by screening all possible points of entry and adopting additional measures where appropriate.
Efficient invertebrate vector control is easier to achieve in a glasshouse or growth room, than a polytunnel, by:
- adopting the limited use of mechanical/manual ventilation systems,
- screening of all air inlets and exhausts to a suitable size for the vectors concerned,
- using a vestibule/lobby arrangement to enter the quarantine station/confinement facility,
- fitting brushes or rubber strips around all doors and
- using regular, appropriate chemical control regimes and traps (e.g. yellow sticky traps for aerial insect vectors and sticky foot traps or footbaths containing disinfectants at the exits).
Containment of invertebrates or invertebrate vectors of pathogens
It is likely that the containment of invertebrates in association with plants, or the experimental transmission of specified plant pathogens using invertebrate vectors, will only be permitted in dedicated growth cabinets/rooms or insectary facilities. This may include additional mitigations, for instance the use of thermal or light gradients or negative pressure for the containment of mobile invertebrates.
The work carried out should preferably involve the minimum number of plants, be of short duration, and be undertaken when the environmental conditions outside the facility are less likely to permit the survival of the invertebrate vector. The construction of a specialist insectary facility containing growth cabinets in which plants can be grown is encouraged, with the application of temperature and light gradients providing additional barriers to invertebrate movement.
Containment of plant pollen and seeds, including when infected with plant pests and pathogens
Suitable procedures should be in place to prevent the dispersal or escape of any plant pollen and seed associated with material being held under a scientific authorisation. In cases where it is not necessary for pollination to take place, this can be easily achieved by disbudding the flowers or by terminating the experiments prior to flowering. In cases where it is necessary for pollination to take place, other measures should be adopted such as
- the bagging of flowers,
- the use of additional caging,
- the control of invertebrate pollination vectors,
- the filtering of exhaust air outlets to a size appropriate for the pollen in question,
- the use of the minimum number of plants required and
- the spatial and seasonal isolation of the plants from sexually compatible species.
Transfer of material around the quarantine station/confinement facility
The procedures involving the transfer of specified material around and between approved quarantine stations/confinement facilities should limit the risk of escape of the specified material or pests.
Transfer of specified material between approved facilities on site must be kept to a minimum, but where transfer is unavoidable specified organisms or material should be transported within three layers of containment. This should involve the use of sealed bags and/or closed, sealed, shatterproof containers as appropriate.
Inoculation methods
The main consideration with inoculations is whether they produce aerosols, and how those aerosols could be contained.
Inoculations creating aerosols or run off include:
- Spray inoculations
- Dip inoculations
- Mesophyll inundation (forcing under pressure through a syringe into a leaf)
- Agrobacterium-mediated infection
Inoculations not creating aerosols or run-off include:
- Agar plug inoculations (usually used for non-sporulating fungi)
- Slit inoculation
- Rub inoculations (used for viruses)
- Germination in a culture/sand mix (used for bacteria)
Both aerosols and surface run off can be contained within a MSC II effectively. However, there may be alternatives (for instance if plants are too large to work on in a MSC II). For example, Inoculation techniques which produce surface run off (e.g. dip inoculations) could be performed in a tray, with the run-off contained effectively and then sterilised. Whilst with spray inoculations these could be performed outside a MSC II, provided certain conditions are met, such as:
- The growth room/(s) where inoculations take place have negative pressure and HEPA filters are in place.
- A polybag, polythene covered cage or plastic box is put over the plant prior to inoculation and is sealed around the plant (as much as possible) and left for a minimum period of 24 hours, to minimise aerosol release.
- Lab coats are left within the room used for inoculations.
- Any material in the growth room is disinfected or autoclaved after the inoculation has taken place or the experiment has ended.
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