HeV sampling equipment and procedures

Preferred samples

1. Nasal, oral (tongue surface), and rectal mucosal (not faecal) swabs.
2. Blood – 1x 10ml serum tube, 1x10ml EDTA tube.
3. Urine swab, taken from the ground immediately post-urination if possible.

Sampling equipment for each live horse

• shielded vacutainer needle and holder, plus several spares
• 1x serum tube, 1x EDTA tube, plus spares
• 3 x virus transport media (VTM), plus spares – if VTM is not available, place swabs in small amount of saline
• 4 x swabs, plus spares
• sharps disposal container.

Preferred samples

1. Nasal, oral, rectal mucosal swabs.
2. Blood, if available – 1x serum tube, 1x EDTA tube.
3. Additional necropsy samples:
   • blood clot taken from cutting the jugular vein
   • submandibular lymph node tissue.

Sampling equipment for each dead horse

• shielded vacutainer needle and holder, plus several spares
• 1x serum tube, 1x EDTA tube, plus spares
• 3 x VTM, plus spares
• 4 x swabs, plus spares
• scalpel, scissors, and forceps to collect tissue samples
• sample jars for fresh tissue samples in formalin
• cut-resistant gloves (Kevlar) may be considered.

PPE for each person

• disposable overalls with at least splash resistant rating, plus a disposable or washable hat if the overalls have no hood
• one of the following respirator options:
  • disposable P2 or N95 particulate respirator, plus spares
  • reusable negative pressure respirators
  • powered air purifying respirator (PAPR) and filters
• pack of disposable gloves – nitrile gloves recommended
• safety eyewear or a face shield
• roll of duct tape
• pair of impervious rubber boots.

Respiratory requirements

We recommend discussing specific respiratory protection needs with a supplier of safety equipment.

The minimal level of respiratory protection when investigating a potential HeV case is P2 particulate respiratory protection.

Disposable particulate respirators must be fitted correctly and are not suitable for people with facial hair. These people should use a powered air purifying respirator (PAPR) which draws air through a filter and supplies it to a hood worn over the head.

Overalls

Splash-proof overalls are lighter and suited to hotter conditions that may be faced during investigation.

Impervious overalls present a risk of overheating in less than 20 minutes, particularly if used in direct sunlight.

Waste disposal

• small sharps container with built-in needle removal facility
• 2x A4 clip seal bags to remove samples from premises
• plastic bucket for carrying equipment
• 1x 500 ml spray pack of Virkon, or other approved disinfectant.

Disinfection

• foot bath and 2 or 3 buckets
• scrubbing brush
• hoof pick or medium screwdriver
• 20 litres water
• disinfectant sachets or bulk supply for mixing at 50g per 5L water, such as Virkon or Trigene
• soap or detergent
• small hand sprayer for chosen disinfectant – if using reusable respirators
• 1x heavy duty garbage bag, plus 1x clinical waste bag
• 2x zip or cable ties
• ground sheet or plastic mat, no more than 1 square metre.

HeV is a member of Category A viruses, which contain a lipid envelope.

The following disinfectants are recommended for either their effect against all viruses, or their specific action against Category A viruses. Use these when handling horses that may have HeV:

• soaps and detergents
• Virkon
• hypochlorites
• iodophors, iodine
• biguanidines such as chlorhexidine
• quaternary ammonium compounds.

Other disinfectants are considered effective, but they require special precautions for safe use. See the AUSVETPLAN decontamination manual for further information.

Enter the dirty area only after you are fully dressed in PPE and have all the required gear ready. Follow this sequence:

1. Wash hands with soap or detergent.
2. Put on overalls then boots – overall legs go outside the boots.
3. Put on safety eyewear, gloves and disposable respirator – if using a PAPR instead, put this on, along with gloves, after step 5.
4. Fit check the respirator.
5. Pull overalls hood up if present and zip to chin – if the overall have no hood, put on a protective hat.
6. Double-glove and tape the outer gloves onto the sleeves of the overalls.

Anyone assisting you, or nearby, must wear the same standard of PPE.

It is critical to minimise the chance of contaminating people and their PPE. Follow these precautions when taking samples:

1. Clearly and uniquely label all specimens.
2. Undertake safe sharps handling and waste disposal – use the sharps container and do not re-cap needles. If you're accidentally exposed to blood or body fluid, or sharps injury:
   • Wash the affected area of skin thoroughly with soap and water, or irrigate mucous membranes with water or saline.
   • Seek immediate medical advice.
3. Once sampling is complete, place in a clip seal bag for removal.

Avoid spreading contamination

Follow these steps before entering the designated clean area:

1. Remove gross contamination from yourself and equipment before reaching the entry and exit point. Use a brush and soap, or detergent and water.
2. Clean the treads of the boots with a tap on site, or the bucket in the transition area.
3. Go to the dirty side of the transition area to double-bag the samples in clip seal bags.
4. Disinfect the samples on the clean side – be careful not to contaminate them with disinfectant.
5. Spray disinfectant on the outer gloves.

Depending on the type of respirator used, there are different procedures for removal, as below:

1. Remove the outer pair of gloves to garbage bag.
2. Wash hands, still encased in the inner pair of gloves, in disinfectant.
3. Remove boots and stand on drop sheet in the transition area.
4. If using a P2 respirator or a negative pressure full-face respirator, then:
   • Remove overalls to garbage.
   • If wearing a hat, either remove it to garbage bag, or soak in disinfectant, seal in a bag and remove for laundering.
   • Remove and disinfect safety eyewear.
   • Remove respirator and either throw disposables in garbage, or wipe reusable respirators with disinfectant solution.
   • Remove inner gloves to garbage.
5. If using a PAPR, then:
   • Remove and disinfect the PAPR.
   • Remove overalls to garbage.
   • Remove the inner gloves to garbage.

Decontaminate equipment

Take care not to re-enter the dirty area while disinfecting equipment and follow these steps:

1. Place all waste material in garbage bag, seal it with zip tie, and disinfect surface.
2. Double-bag waste by placing garbage bag into a clinical waste bag, seal with zip tie, and disinfect surface.
3. Disinfect and double-bag non-disposable items for removal.
4. Put on street shoes.
5. Pack up disinfection site, disinfecting all equipment thoroughly as packed.
6. Wash disinfectant off reusable respirators with clean water.
7. Wash hands and other exposed skin with soap and water.

If non-disposable PPE cannot be adequately decontaminated on site, double-bag it and remove for later attention – this is not a preferred option.

Before leaving the property, discuss these necessary biosecurity matters:

1. Enquire as to the amount of human contact people on the property have had with the sick or dead horse.
2. Advise the property owners or managers on:
   • appropriate biosecurity processes and PPE
   • isolating sick or dead horses from people and other animals, including pets
   • limiting movement of horses, horse products, and visiting practitioners on and off the property
   • disposing of the horse body on the property – this should only be undertaken by people who are aware of the risks and appropriate methods
   • seeking medical advice if unwell or concerned about possible exposure
   • contacting SA Health's Communicable Disease Control Branch if needed.
3. Inform the Biosecurity division of the suspect Hendra case, or provide an update of situation if already notified.

If possible, avoid any risks of spreading HeV until the results are known. This includes:

• contact with other animals and people after leaving the property
• using contaminated equipment that was taken into the dirty area, which should have been decontaminated and double-bagged in the transition area.

If the equipment cannot be isolated until the HeV test results are returned, wear PPE to clean it, using detergent then disinfectant. Be careful when removing the PPE.

If the results are negative, clean equipment per the standard practice routine. If results are positive, contact PIRSA's Biosecurity division.