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SAFETY & HEALTH REQUIREMENTS FOR USE OF MATERIALS DERIVED FROM NHPS

Introduction

1. Experimentation with NHP-derived samples (cell cultures, tissues, body fluids or other wastes) may

expose NUS staff and students to various biohazards. Of all these biohazards, the risk of Herpes B virus

(Cercopithecine herpesvirus I, Herpesvirus simiae) infection (zoonosis) from working with macaque-

derived samples is of particular concern because the fatality rate in untreated human B virus infection

has been reported to be greater than 70% (Rohrman, 2016).

2. NUS therefore requires BSL2 containment for any staff and students working with NHP-derived

samples that have not undergone processes to inactivate/denature potential infectious agents.

Researchers using macaque-derived materials are required to work in a BSL 2 enhanced laboratory.

These are laboratories where additional practices and controls are in place in addition to Universal

BSL2 requirements. The requirements and controls for working with different NHP materials are given

in the table below:

Type/ Hazard NUS Required Risk controls

References

Macaque-derived materials*

BSL2 enhanced laboratory. In addition to Universal BSL2 precautions, the PI shall ensure the following:

a. Obtain approval from IBC prior to commencement of any work on or possession of macaque-derived materials

a. Applications for approval are done via the iORC. (https://inetapps.nus.edu.sg/osh/portal/eServices/iorc.html).

b. Staff and students participate in the

NUS OH programme that has been developed for handling macaque-derived materials.

b. More information about the enrolment requirements and procedures can be found in the following link –“NUS OH Programme for Personnel with contact to Laboratory Animals.”

c. Staff and students complete the

classroom based training course

entitled ‘Safe Handling of Non-

Human Primate (Macaque) Derived

Materials’ conducted by OSHE.

c. Procedure for registration for this course is found at NUS SSTS page.

d. Procedures are in place to ensure restricted access to authorized

d. Access to such areas can be managed by the use of card-

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personnel for areas where macaque-derived material are handled.

access systems or by other security means.

e. Adequate signage is displayed at locations where macaque-derived material is handled. It is a requirement for the signage to contain the following terms: ‘Macaque Monkey-Derived Material in Use’, and this should be accompanied with a biohazard symbol.

f. Establish a procedure for staff and students to follow in the event of any exposure to macaque derived materials. The laboratory shall set up a NHP exposure kit. Requirements of the NHP exposure procedure and kit are given in Annex B.

e. Refer to ANNEX A for a sample f. Refer to ANNEX B for the

requirements, for the NHP exposure kit contents and of the NHP exposure response procedure

NHP (other non-macaque) derived materials*

BSL2

Inactivated/ Denatured NHP derived materials**

BSL1

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ANNEX A: Additional Hazard Communication Signage

MACAQUE MONKEY-DERIVED MATERIALS IS USED

IN THIS LAB

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ANNEX B: REQUIREMENTS OF LABORATORY’S POST EXPOSURE RESPONSE PROCEDURE AND

CONTENTS OF THE NHP EXPOSURE KIT

ANNEX B1. The Laboratory post exposure response procedure shall detail the following:

A) Immediate response post exposure

1) For scratches and needle sticks resulting in exposure to macaque derived material.

a. Immediately follow these first aid procedures using the NHP Exposure Kit and BEFORE

proceeding to OH or ER Services at NUH.

i. The kits should be easily accessible and users should be familiar with their

locations.

b. Open NHP Exposure Kit and follow the instructions.

c. Put on exam gloves before scrubbing wound. Ensure gloves are properly worn to

prevent any seepage beneath the gloves.

2) If the injury involves a single hand, glove the other hand before proceeding.

3) If the injury does not involve hands, glove both hands before proceeding.

a. With a gloved hand, scrub the wound vigorously with chlorhexidine-soaked scrub

sponge side (NOT plastic brush side) and water for 3-5 minutes.

b. During cleaning, encourage blood to flow out from the wound by placing pressure

around the area of injury.

c. Irrigate the washed area with running water for at least 15 minutes

d. Discard sponge into biohazard waste bags/ containers

e. Cover the wound with sterile gauze, if appropriate.

f. Collect Packet of Documents and Sample Collection Supply from the NHP Exposure kit.

Take them with you to Occupational Health Clinic (OH)/ NUH Emergency Medicine

Department (NUH ER).

4) For eye splashes and mucous membrane exposures

a. Immediately follow these first aid procedures using the NHP Exposure Kit and BEFORE

proceeding to OH or ER Services.

i. The kits should be easily accessible and users should be familiar with their

locations.

b. Open NHP Exposure Kit and follow instructions.

c. Wash hands and put on exam gloves.

d. Rinse eyes or exposed mucous membranes immediately with water at an eye wash

station or sink for 15 minutes.

e. If water/ eye wash station is not immediately available:

i. Use ophthalmic irrigating solution (found in the NHP Exposure Kit) and make

your way to the nearest eye wash station/ sink.

ii. Continue to rinse eyes or exposed mucous membranes at an eye wash station

or sink to allow total rinse of 15 minutes.

f. DO NOT USE CHLORHEXIDINE IN THE EYES.

g. Collect Packet of Documents and Sample Collection Supply from the NHP Exposure kit.

Take them with you to Occupational Health Clinic (OH)/ NUH Emergency Medicine

Department (NUH ER)

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h. Inform your supervisor or another person (who shall inform your supervisor) as soon

as possible of your injury

5) Proceed to OH or NUH ER:

a. During regular clinic operating hours (8.30am- 5.30 pm), the supervisor should call OH

to verify if the physician is in.

Occupational Health (OH)

University Health Centre, Basement

National University of Singapore

20 Lower Kent Ridge Road

Singapore 119080

Dr. Gregory Chan Tel: 6516 5969, Mobile Number: 81129214

Nurse Lee Kim Geok Tel: 65167333

Goh Sha Wee Tel: 6601 1781

b. If the OH physician is not in, or after clinic operating hours (8.30am-5.30pm), proceed to

the nearest Accident and Emergency (A&E) Department near Kent Ridge Campus.

Emergency Medicine Department (NUH ER)

5 Lower Kent Ridge Road

1 Main building, level 1

Singapore 119074

Tel: (65) 6772 6229

24-hour Emergency Department Service Hotline: (65) 6772- 5000.

Research laboratories located outside Kent Ridge Campus to identify the nearest (A&E).

B) Personnel Post Exposure Examination and Sample Collection at OH or NUH ER

1. Take with you the Packet of Documents and Sample Collection Supply from the NHP Exposure

Kit, consisting of:

a. Copy of lab’s post response procedure.

b. First Aid quick reference cleaning protocol card ANNEX B2.1

c. Medical Alert Information sheet ANNEX B2.2.

d. “B Virus Exposure Mini Protocol” ANNEX B2.3.

e. Copy of the National B Virus Resource Laboratory Submission form ANNEX B2.4.

i. OH/NUH ER to complete the form for sample submission.

ii. CM supervisor to fill out animal number, date of injury, and type of injury or

exposure (needle stick, splash etc.).

f. Two (2) plain tubes (3 ml each) for blood serum collection.

g. Sterile swabs and Viral Transport Media (VTM) so a swab for virology can be taken

from the wound or exposed surface.

2. Provide the packet to the physician at OH/ NUS ER.

3. Blood and viral cultures should be obtained from the affected researcher:

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a. Blood samples for serology:

i. Collect two (2) 3 ml samples of blood in red-topped tubes. Do not use

anticoagulants.

ii. Label the tubes with the date, and patient identification.

b. Viral Cultures:

i. Collect swabs sample from affected area.

ii. Use sterile cotton or Dacron swabs with plastic shaft.

iii. Put swab in separate Viral Transport Medium (VTM) and cut the shaft with

sharp scissors (do not break the swab’s shaft by hand).

4. Label each tube with the date, patient identification, and site from which the culture was taken

a. After sample collection, OH clinic staff will take all blood tubes, swab and completed

form back to CM Diagnostic (D-Lab) for processing and for shipment with the NHP

samples to the National B Virus Resource Laboratory. (Please refer to the Biorisk

Management Manual for triple packaging requirements for collected samples)

b. Follow doctor’s instruction and recommendation if B virus prophylaxis is prescribed.

c. The affected person may need to be started on antiviral prophylaxis (Valacylovir, 1 g) for

2 weeks. When that occurs, then the follow-up serum sample for testing would be 2

weeks after the antiviral prophylaxis is completed.

C) Reporting the injury

1) Supervisor of the injured personnel to contact OSHE animal biosafety safety officer within 2

hours at 6601 1169.

2) Supervisor of the injured personnel to contact CM veterinarian who will coordinate with D-

Lab.

3) Report the incident to OSHE via AIMS.

D) NHP Post Exposure Examination, Sample Collection and Submission to D-Lab

1) Obtain a sample of the specimen (NHP derived sample). (Refer to the following link for the

recommendation for sample collection and storage procedures for various sample types.

2) Submit the samples to the CM Diagnostic Laboratory (D-lab) for shipment to the testing

laboratory.

a. If D-lab staff are not available, store samples in D-Lab sample refrigerator.

b. Inform D-lab personnel that the samples are from a macaque so that laboratory

personnel take appropriate precautions.

c. The D-lab will arrange for serology and virus cultures to be shipped.

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ANNEX B2. The minimum contents of the NHP exposure kit shall be as follows:

1. Immediate post exposure response items:

a. First Aid quick reference cleaning protocol card. An example is provided in ANNEX B2.1

b. Chlorhexidine scrub sponge (contains chlorhexidine))

c. Ophthalmic/ saline irrigation solution (for use when emergency eye wash is not available)

d. Disposable nitrile gloves (different sizes)

e. At least 2 small Biohazard bags (to contain potentially contaminated, used solid items)

2. NHP Sample Collection Kit (Packet of Documents and Sample Collection Supply): Sample

collection kit to be brought to the doctor:

a. Materials to keep the sample collection supply and to transport the NHP samples to D-Lab

(Triple packaging supplies)

b. Medical Alert Information Sheet ANNEX B2.2

c. B Virus Exposure Mini Protocol ANNEX B2.3

d. National B Virus Resource Laboratory submission form ANNEX B2.4

e. For blood samples for serology:

i. Two (2) 3 ml samples of blood in red-topped tubes. Do not use anticoagulants.

ii. Syringe (3 ml) and needles (21 or 23 gauge)

iii. Alcohol swab

iv. Gauze

f. For viral cultures:

i. Four (4) sterile Dacron swabs with plastic shaft

ii. Four (4) Viral Transport Medium (VTM)

iii. Scissors to cut the plastic shaft (to fit in the VTM tube).

The kits should be frequently checked to ensure they are fully equipped and all items are usable. Used or expired items shall be replaced as soon as possible. (Refer to the NUS First Aid and First Aiders Standard for more details).

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ANNEX B2.1 First Aid Quick Reference Cleaning Protocol Card

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ANNEX B2.2 Medical Alert Information sheet

Please contact: Dr. Gregory Chan Tel: 6516 5969, Mobile: 81129214;

or OH Clinic - Nurse Lee Kim Geok Tel: 65167333; or Goh Sha Wee Tel:

6601 1781

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ANNEX B2.3 B Virus Exposure Mini Protocol

A

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ANNEX B2.4 National B Virus Resource Laboratory submission form

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ANNEX B2.4 (continued) National B Virus Resource Laboratory submission form

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REFERENCES

1. National B Virus Resource Center - http://www2.gsu.edu/~wwwvir/

2. B Virus Information at Viral Immunology Center –

http://www2.gsu.edu/~wwwvir/VirusInfo/index.html

3. United State Center for Disease Control and Prevention -

https://www.cdc.gov/herpesbvirus/

4. CM SOP #551.03 Procedure for Bites, Scratches and Mucous Membrane Exposures from

Macaque NHPs

5. Vasireddi M and Hilliard J.; Herpes B virus, macacine herpesvirus 1, breaks simplex virus

tradition via major histocompatibility complex class I expression in cells from human and

macaque hosts. Journal of Virology. 2012; 86(23)

6. Cohen JI, Davenport DS, Stewart JA, Deitchman S, Hilliard JK, Chapman LE, B virus Working

Group; Recommendations for Prevention of and Therapy for Exposure to B Virus

(Cercopithecine Herpesvirus 1). Clinical Infectious Diseases 2002; 35(10):1191-203.

7. Rohrman M.; Macacine Herpes Virus (B Virus) Workplace Health & Safety 2016; 64(1):9-12.

8. Weigler BJ.; Biology of B virus in macaque and human hosts: a review. Clinical Infectious

Diseases 1992 Feb; 14(2):555-67.

9. Abee CR, Mansfield K, TardifS, Morris Teds: Nonhuman Primates in Biomedical Research 2nd

ed, Volume 2: Diseases, pgs 7-13