Identify any student or worker who could be … · Web viewBinders containing SOP’s in RANNEX Rm 146 with SOP’s relevant to work performed and RANNEX Emergency Response Plan.

NOTE: This is only an example of a biosafety plan that can be used as a tool when completing your own biosafety plan.

Also, refer to the Biosafety Plan Guide for further instruction and examples on completing each section in the plan.

For any assistance or questions, please contact the Biosafety group: [email protected] .

mailto:[email protected]

Biosafety PlanYear: 2019

Department: Safety Resources

Permit Holder Name:Dr. Norman Legacy

Biosafety Permit Number:SFT-01

Contents

1 Contact Information....................................................................................................................1

2 Nature of Research................................................................................................................2

3 Biological Assessment and Inventory.....................................................................................3

4 Health and Safety Hazard Assessment..................................................................................5

5 Biosecurity Assessment.........................................................................................................8

6 Work Locations.....................................................................................................................10

7 Standard Operating Procedures (SOPs) and Element Requirements.................................12

8 References...........................................................................................................................17

Revision History

Revisions to the biosafety plan are documented in Table 1, Revision History (e.g. New Biosafety Plan, Addition of new biohazardous materials, etc.).

Table 1: Revision HistoryDocument

Section Details of Amendments Date Author(Initials)

1 New level 2 biosafety permit January 2018 NL

2 Added wild-type and recombinant porcine epidemic diarrhea virus (PEDV) June 2019 NL

**For new amendments, please highlight the revised portions of the Biosafety Plan.**

1 Contact Information

Refer to Section 1 of the Biosafety Plan Guide.

Biosafety Permit Holder Information

Biosafety Permit Number: SFT-01

Biosafety Permit Issue Date: January 17, 2018

Permit Holder Name: Dr. Norman Legacy

Permit Holder Telephone Number: 306-966-6666

Lab Telephone Number: 306-966-8888

Email: [email protected]

Secondary Contact Name: Jiff Smith

Secondary Contact Telephone Number: 306-966-7777

Email: [email protected]

Emergency Contact Information

Primary Contact:Name: Dr. Norman Legacy

Office Phone Number: 306-966-6666

Cell Phone Number: 306-555-5558Secondary Contact:Name: Jiff Smith

Office Phone Number: 306-966-7777

Cell Phone Number: 306-552-2222Tertiary Contact:Name: n/a

Office Phone Number: n/a

Cell Phone Number: n/a

Safety ResourcesMay 2019 1 | P a g e

2 Nature of Research

Objective #1: To compare vaccinated and non-vaccinated swine when naturally exposed to porcine respiratory virus (PRV).

Methods Used: Use tissue culture methods and real-time polymerase chain reaction (PCR) on tracheal

mucus samples obtained from the swine to measure target protein production. Use real-time PCR and tissue culture methods to determine what percentage of swine

who become infected with PRV; and Use of immune-histo techniques to compare the proportion of and days to disease

development between vaccinated and non-vaccinated heifers as determined by ultrasonography and tracheal glycoproteins.

Objective #2: To develop a vaccine against the porcine epidemic diarrhea virus (PEDV) by identifying the various virulent and variant strains found in North America, and determining if the variant strain can be used as a vaccine. Once a virulent strain is determined, the infectious nature of the disease will be tested on pigs, to determine symptoms.

Methods used: Collecting various strains of PEDV across North America, including Canada and the

USA; Using cell culture to propagate the PEDV, this will be confirmed by immunofluorescence

assays and electron microscopy; and Using real-time polymerase chain reaction (PCR) to identify the genetic differences

between PEDV strains; and validate the use of real time PCR process for identifying the genetic differences.

Pig studies to determine virulence factors, taking samples, blood, and determining vaccine potential targets.


3 Biological Assessment and Inventory

Table 2: Inventory of Biological/ Biohazardous Material.

Name of Biological Material

1

Supplier/Source of the

Biological Material 2

Purpose or Use 3

Concentration 4

Max. quantity to be cultured at one time?

(in liters)

Human Risk Group 5

(1, 2, 3)

Animal Risk

Group 5

(1, 2, 3)

Containment Level 5

(1, 2, 3)

Location

(room, freezer numbe

r)

rDNA/GMMO 6 (Yes or No)

Dual-use Potential 7

(Yes or No)

Reference(s) 8

Porcine respirator

y virus (PRV)

ATCC CRL-1234

Research in vitro and storage

5x107/ml PFU,1-10 µl 100 mL 1 2 2

WCVM

Room 1,

Freezer 2

No Nowww.attc.com/CRL-

123

Genetically

Modified PRV

Oklahmoma State, USA

Research, in vitro and storage

5x107/ml PFU,1-10 µl 100 mL 1 2 2

WCVM

Room 1,

Freezer 2

Yes Nowww.attc.com/CRL-

123

Porcine cell line (A13)

ATCC CRL 4589


106 cells/mL <250 mL 1 1 1

WCVM

Room 1,

Freezer 2

No Nowww.ATC

C.com/CRL-4589

Porcine cell line (Be2)

(contains adenovir

us compone

nts)

University of Kentucky,

USA


106 cells/mL <250 mL 2 2 2

WCVM

Room 1,

Freezer 2

Yes Nowww.atcc.com/CRL-

1458

Animal tissues

and blood

(healthy)

Prairies Swine Centre


n/a Blood: <100 mL

Tissues: samples

only

1 1 1 WCVM

Room 1,

Freez

No No n/a


http://www.ATCC.com/

http://www.ATCC.com/


1



Purpose or Use 3

Concentration 4


(in liters)

Human Risk Group 5

(1, 2, 3)

Animal Risk

Group 5

(1, 2, 3)

Containment Level 5

(1, 2, 3)

Location


r)



(Yes or No)

Reference(s) 8

- Porcine er 2

Human cell line (M11)

University of Waterloo, Canada


106 cells/mL <500 mL 2 1 2

WCVM

Room 1,

Freezer 2

No No

Exp. Cell Research, 2017: 204-222, 1998.

Human cell line (PHM-

43)

University of Waterloo, Canada


106 cells/mL <500 mL 2 1 2

WCVM

Room 1,

Freezer 2

No No

Devost D., and Zigg

LH, Am. J. Physiol. 295: D1-D8, 2009.

Recombinant DNA

(plasmids and

expression

vectors)

AddgeneResearch, in

vitro and storage

52 stocks of 2 mL each; Conc. ~ 1

mg/mL

50 mL 1 1 1

WCVM

Room 1,

Freezer 2

Yes No Refer to rDNA form

Escherichia coli DH10B carrying pET41a(

+)

Harvard Medical

School ,USA


various from 1X102 to 1 X109 cfu/ml

if liquidor bacterial growth on plates cfu

not enumerated

500 mL 1 1 1

WCVM

Room 1,

Freezer 2

Yes No

SDS Invitrogen: httpss://tools/Invitrogen.com/content/sfs/sds/2010/18265017_MTR

-NAIV_EN.p

df Live

animals, pigs

Prairie Swine Centre

In vivo n/a n/a 1 1 1 WCVM

Room

No No AUP# 201900458



1



Purpose or Use 3

Concentration 4


(in liters)

Human Risk Group 5

(1, 2, 3)

Animal Risk

Group 5

(1, 2, 3)

Containment Level 5

(1, 2, 3)

Location


r)



(Yes or No)

Reference(s) 8

(healthy)1,

Freezer 2

Porcine epidemic diarrhea

virus (PEDV)

Dr. Klaus Osterrieder, Dept. of Veterinary Medicine, Freie Universität, Berlin

In vitro, in vivo and storage

5x107/ml PFU,1-10 µl 250 mL 1 2 2

WCVM

Room 1,

Freezer 2

No No

https://www.ncbi.nlm.nih.gov/

pmc/articles/

PMC5799402/

Genetically

modified PEDV

(please see

rDNA-GMMO-

transgenics Form for more detail)

Dr. Klaus Osterrieder, Dept. of Veterinary Medicine, Freie Universität, Berlin

In vitro, in vivo and storage

5x107/ml PFU,1-10 µl <250 mL 1 2 2

WCVM

Room 2,

Freezer 3

Yes No

https://www.ncbi.nlm.nih.gov/

pmc/articles/

PMC5799402/

1 When identifying the biological material, ensure to state the most descriptive name (e.g. genus species) and include the strain if it is a bacteria or virus. No2 State how the biological material(s) is acquired, or procured, such as naming the supplier and their location (e.g. Dr. Smith, University of Brisbane, Tawoomba, Australia). 3 State the type of work which will be conducted with the biological material (e.g. in vitro, in vivo, both, storage only, etc.).4 State the concentration of the biological material (e.g. cells: 106 cells per plate; bacteria: colony forming units; virus: plaque forming units/tissue culture infectious does; toxins: unit of mass/unit of volume);5 The identified risk group (human and/or animal) and containment level as specified by external sources, or as determined from a biological risk and containment level assessment.6 If the biological material is identified as rDNA/GMMO, complete the Risk Assessment for rDNA/Genetically Modified Organisms (GMO) Form.7 Use the Dual-use assessment flow chart in Figure 2 in Section 3 of the Biosafety Plan Guide.8 Indicate the reference(s) for the categorization of the risk group and containment level (e.g. risk assessment performed by researcher, PHAC, CFIA).


https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5799402/






4 Health and Safety Hazard Assessment

Table 3: Hazards Assessment

NOTE: List all biologicals agents listed in the inventory table 2. Similar biological agents, such as cell lines, can be grouped together if the processes are the same. See examples below.

Biological Materials 1Procedures Hazard Assessment

List of Procedures using Biological Material

Potential Route(s) of Exposure from Procedures Safety Mitigation Controls

PRV and genetically- modified PRV

Pipetting, mixing, sonicating, or vortexing biohazardous material

Ingestion, inhalation, and/or absorption through mucous

membranes

- Wear proper PPE (safety glasses, lab coat, disposable gloves)

- Follow specific procedural SOP - Ensure proper training has been

received- Use a biosafety cabinet for pipetting

and mixing- Wash hands with soap and water

for 20 seconds- Follow good microbiological

practicesUse of appropriate disinfectant

Centrifuging human or animal blood, fluid, tissue, organisms or

biohazardous material

Inhalation and/or absorption through mucous membranes


- Follow specific procedural SOP - Use of a sealed rotor cup - Ensure proper training has been

received- Use a biosafety cabinet to open the

sealed rotor cup- Wash hands with soap and water

for 20 secondsFollow good microbiological practices

Handling tubes or other containers Inhalation (only if Cryovial - For Cryovials: use thermogloves Safety ResourcesMay 2017 6 | P a g e

of human or animal blood, fluid, tissue, or other biohazardous

material

explodes, or opened outside of BSC), absorption through

mucous membranes, and/or ingestion

and faceshield when placing or removing vials from liquid nitrogen



received- Use a biosafety cabinet- Wash hands with soap and water


practices

Real time Polymerase chain reactions


- Propagating in porcine and human cell lines

- Wear gloves, safety glasses, and lab coats

- Utilize good microbiological practices

- Utilize the use of the biosafety cabinet

- Work surfaces disinfected with 70 % ethanol after use with a contact time of 5 minutes

Waste disposed in biohazardous waste container

Porcine cell line (A13) and Porcine cell line (Be2) (contains adenovirus

components)

and

Human cell lines (M11 and PHM-43)

Pipetting, mixing, sonicating, or vortexing human or animal blood,

fluid, or tissue


membranes






practices


- Waste disposed of biohazardous waste container









practices- Waste disposed of biohazardous

waste container

Cell culture for RNA, DNA, protein or extracellular vesicle isolation


membranes




sealed rotor cup- Waste disposed of biohazardous

waste container- Wash hands with soap and water


practices- Work surfaces disinfected with 70 %

ethanol after use with a contact time of 5 minutes

Animal tissues and blood (healthy) - Porcine

Handling human or animal tissue, including preparation, dissection,

cutting

Inoculation or/and absorption through mucous membranes


- Use of safety engineered SHARPs


- Use of proper SHARPs container- Follow specific procedural SOP - Ensure proper training has been



practicesPipetting, mixing, sonicating, or

vortexing human or animal blood, fluid, or tissue


membranes






practices- Work surfaces disinfected with 70 %










practices - Work surfaces disinfected with 70 %



Escherichia coli DH10B carrying pET41a(+)

To propagate plasmids and expression vectors

Absorption through mucus, ingestion, and inhalation



- Materials used with these techniques such as agar plates, disposable plastic culture tubes, and plastic centrifuge tube are collected in biohazardous waste

- Glassware washed prior to normal cleaning


Recombinant DNA (plasmids and expression

vectors) (Level 1)

Propagating cDNAs for use in the production of probes for molecular

biological experiments (eg. RT PCR)

Absorption through mucus and skin (e.g. scratches)



- Materials used with these techniques such as agar plates, disposable plastic culture tubes, and plastic centrifuge tube are collected in biohazardous waste

- Glassware washed prior to normal cleaning


Expression of proteins in established cell lines and molecular

biological experiments (e.g immuno- histo assays)

Absorption through mucus and skin (e.g. scratches); inhalation



- Utilize the use of the biosafety cabinet



- Waste disposed in biohazardous waste container

Live animals, pigs

Injecting vaccine, drugs, euthanizing agents, etc. into

animals

Inoculation, inhalation - Wear proper PPE (safety glasses, lab coat, disposable gloves)

- Use of safety engineered SHARPs- Use of proper SHARPs container- Follow specific procedural SOP - Use proper restraints (if required)

Ensure proper training has been provided

Handling contaminated sharps or other biohazardous waste

Inoculation, ingestion, and/or absorption through mucous

membranes



received- Wash hands with soap and water

for 20 secondsFollow U of S Hazardous Waste

Disposal StandardConducting surgical techniques on

animalsInoculation - Wear proper PPE (safety glasses,

lab coat, disposable gloves)- Use of safety engineered SHARPs- Use of proper SHARPs container- Follow specific procedural SOP - Use proper restraints (if required)

Ensure proper training has been received

Handling of live animals – healthy and infected; husbandry

Spray in mucosal membranes, inoculation, inhalation

-

Porcine epidemic diarrhea virus (PEDV) and genetically

modified PEDV

Handling tubes or other containers of human or animal blood, fluid, tissue, or other biohazardous

Inhalation (only if Cryovial explodes, or opened outside of

BSC), absorption through

- For Cryovials: use thermogloves and faceshield when placing or removing vials from liquid nitrogen


material mucous membranes, and/or ingestion





practicesCentrifuging human or animal

blood, fluid, tissue, organisms or biohazardous material







practicesHandling tubes or other containers

of human or animal blood, fluid, tissue, or other biohazardous

material

Inhalation (only if Cryovial explodes, or opened outside of

BSC), absorption through mucous membranes, and/or

ingestion

- For Cryovials: use thermogloves and faceshield when placing or removing vials from liquid nitrogen





practicesPorcine epidemic diarrhea

virus (PEDV) and genetically- Real time Polymerase chain Inhalation and/or absorption - Propagating in porcine and human


modified PEDV reactions through mucous membranes

cell lines - Wear gloves, safety glasses, and

lab coats- Utilize good microbiological

practices- Utilize the use of the biosafety

cabinet- Work surfaces disinfected with 70 %


- Waste disposed in biohazardous waste container

1 Identify and list all Biological materials used in research program (Refer to biological material inventory listed in Table 2).

Table 4: Biological Material Pathogenicity Assessment.

NOTE: - Only include Risk group 2 or higher biological materials for Table 4. - Ensure both the human and/or animal perspective is included in the pathogenicity assessment. - Risk group 1 biologicals materials are not required to be listed in Table 4


Biological Materials 1

Affects Human or Animal or

Both (include

spp.)

Routes of Exposure

2 List of Disease(s) 3 Symptoms 4

Time After Exposure

before Onset of Symptoms

PRV and genetically-

modified PRV

Human n/a Causes no human diseases No human disease n/a

Animal (Pigs) only

Inhalation Pneumonia, meningitis, Infection in the nasal passage Within 5 days (if develops)

ingestion Ingestion

Gastroenteritis, staphylococcal food intoxication from enterotoxins

Nausea, vomiting, abdominal pain,

cramps, and diarrhea

Porcine cell line (Be2) (contains

adenovirus components)

Human

inhalationAdenoviral

components,

Inflammation response, rednessRefer to PSDS for adenovirus:

https://www.canada.ca/en/public-health/services/laboratory-biosafety-biosecurity/pathogen-safety-data-sheets-risk-

assessment/adenovirus-types-1-2-3-4-5-7-pathogen-safety-data-sheet.html

Acute and chronic

Absorption through mucosal

membranes

Adenoviral components Cause inflammation of injection area Acute and chronic

IngestionAdenoviral

components, No known symptoms Acute and chronic

Animal n/a Causes no animal diseases No animal disease n/a

Human cell lines (M11 and PHM-43) (cancer cell

lines)

Humans Mucosal membrane (skin, eyes

Blood born pathogens (Hepatitis B &C, HIV, West Nile virus, Zika

virus)

Hepatitis B & C: Fever, Fatigue, loss of appetite, Nausea, Vomiting, Joint pain, Jaundice, abdominal pain.

HIV: Headache, Diarrhea, fatigue, nausea and vomiting, aching muscles

West Nile Virus: fever, headache, body aches, skin rash, and swollen lymph nodes. Severe symptoms and signs may include stiff neck, sleepiness, disorientation, coma, tremors, convulsions, and

paralysis

Refer to Public Health Agency of Canada’s Pathogen Safety Data Sheets (Refer to PSDS)

Hepatitis C: 2-12 weeks

Hepatitis B: 2-6 months

HIV: 2-6 weeks

West Nile: 3-14 days

Inoculation

Tumours On site of injection – inflammation response, redness Unknown – risk of


https://www.canada.ca/en/public-health/services/laboratory-biosafety-biosecurity/pathogen-safety-data-sheets-risk-assessment/adenovirus-types-1-2-3-4-5-7-pathogen-safety-data-sheet.html




Affects Human or Animal or

Both (include

spp.)

Routes of Exposure

2 List of Disease(s) 3 Symptoms 4

Time After Exposure

before Onset of Symptoms

tumor formation


Porcine epidemic diarrhea virus (PEDV) and genetically-

modified PEDV

Human n/a Causes no human diseases No human disease n/a

Animal (Pigs) only

Inhalation Mild case of upper bronchial infection, Runny nose, redness around nose area, sneezing 1 to 3 weeks

ingestion Ingestion

Gastroenteritis, staphylococcal food intoxication from enterotoxins

Rapid onset, Nausea, vomiting, abdominal pain,

cramps, and diarrhea

Live animals (Pigs)

Human

Inhalation Allergic reaction Respiratory Tract irritation Acute and/or chronic

Absorption through skin

and mucosal

membranes

Allergic Reaction Skin and eye irritation Acute and chronic


1 Identify and list all Biological materials used in research program (Refer to biological material inventory listed in Table 2).2 State the route of exposure which may occur when working or storing of the biological material: ingestion, inoculation (e.g. needle stick injury, bites and scratches), inhalation, and absorption through the skin and/or mucosal membranes.3 List all potential diseases and/or allergies/sensitization that can or may be caused by the biological material (e.g. Human cell line HEK293T cells – cells contain adenoviral components, and risk of Blood borne pathogens, such as Hepatitis, HIV, other blood borne diseases)4 List the symptoms that can be obtained from each route of exposure from the biological material. If a TOXIN is used, state the rate of action for the toxin (how long after exposure before effects of the toxin are observed) and state the LD50 for humans (if no available provide information for the animal).

Table 5: Medical Surveillance and Immunization Assessment. 1

NOTE: - Include only Risk group 2 or higher biological materials in Table 5.


- Include Risk group 1 biologicals materials that can cause allergens in Table 5.- This table is to be completed from the human perspective


Vaccinations 3 Prophylaxis 4 Medical Surveillance 5 Compromised Immune system Allergies

Available

(yes or no)

Effective

(yes or no)

Vaccination Waiver Needed

(yes or no)

Available

(yes or no)

Type of Prophylaxi

s

Serum Collection

Recommended for Baseline

Reference Needed

(yes or no)

Annual Medical

Monitoring

Required (yes or

no)

Are individuals with compromised

immune systems at risk for developing

disease? (yes or no)

Can individuals develop an

allergy (sensitized)

to the biological material?

(yes or no)PRV and

genetically modified PRV

n/a n/a n/a n/a n/a n/a n/a n/a n/a

Porcine cell line (Be2) (contains adenovirus

components)n/a n/a n/a Yes Antivirals n/a n/a Yes n/a

Human cell lines (M11 and PHM-43) (cancer cell lines)

Yes (Hepatiti

s B vaccine)

Yes (Hepatiti

s B)n/a Yes Antivirals n/a n/a Yes n/a

Porcine epidemic diarrhea virus (PEDV)

and genetically modified PEDV

n/a n/a n/a n/a n/a n/a n/a No n/a

Live animals (Pigs) n/a n/a n/a n/a n/a n/a n/a No Yes

1 Depending on the biological materials used, more information may be required by the Biosafety Group.2 Identify and list all Biological materials used in research program (Refer to biological material inventory listed in Table 2).3 If a vaccination is declined, have the individual fill out the Vaccination Waiver form, which can found at the Safety Resources website: http://safetyresources.usask.ca.4 State the prophylaxis, or action taken to prevent disease, such as antibiotics, antiviral agents, antifungal, etc.


http://safetyresources.usask.ca/

5 Medical surveillance, which is the identification of individuals exhibiting signs or symptoms of illnesses or diseases, are proactive exposure-specific initiatives that monitor the effectiveness of workplace safety precautions over time. Medical surveillance may be used to monitor the health of those who are working with potential exposure to blood borne pathogens, hepatitis C, tuberculosis, etc.


5 Biosecurity Assessment

The development, implementation, evaluation, and maintenance of a biosecurity plan, based on a biosecurity risk assessment, is required for facilities where biohazardous material or toxins are handled and/or stored.

Answer the following questions in detail to develop your biosecurity plan.

Part A: Biosecurity PlanWho is allowed, or authorized in the facility (e.g. graduate students, technicians, other faculty, custodial staff, building service technicians, Operations and Maintenance trades, etc.)?

Individuals listed as Authorized Workers on the Biosafety Permit are allowed to work in laboratories listed on the Permit and will have keys/codes to access these laboratories. Custodial staff, building service techs., and Operations and Maintenance trades are permitted into laboratories to perform their required tasks only. Visiting students, faculty, and scientists require full supervision and need to sign in/out log sheet. No children or pets are allowed in laboratories.

Have you identified the roles and responsibilities for each authorized individual? Where is this documented?

Authorized individual roles and responsibilities are documented and kept in the Safety Training Records binder in Rm 163 Research Annex.

Describe how is access to the facility controlled?

Only those authorized to enter the facility are assigned keys for access. Each individual has keys with unique numbers, which are monitored by the Safety Resources managerial clerical staff.

Are doors to the containment zone are kept closed at all times (no exceptions)?

Doors are kept closed at all times. Doors are kept locked at night and when they are left unattended during the day.


How is access monitored in the facility?


Doors are kept locked when unattended to prevent unauthorized access as well as halls monitored by everyone, who report any suspicious activity when detected. All -20 oC freezer stored in the hallways are locked. Only authorized personnel have key access.


Are visitors supervised and sign a log book, which includes name, date and time-in and time-out, purpose of visit?

Yes, visitors are supervised at all times and signed in and out in a logbook along with the purpose of the visit.

Is the intellectual property, including lab books, electronic files, etc. secured and how is this done?

Lab books stay in secured laboratories and electronic files are stored in computers which are password protected. A shared laboratory drive will be used to backup data and IP.

Who is responsible to verify and check the biological and biohazardous material inventory? How often is the inventory updated?

Dr. Legacy verifies inventory updated by the laboratory Tech, currently Jenny Liang. Inventory will be updated every 6 months or more often as needed.

List the individuals who has access to the biological and biohazardous material inventory and documentation.

- Dr. Norman Legacy (PI)- Jeff Smith (Lab Technician)- Mehan Grouse (Master’s student)- Stewart Little (Post doc.)

If the biological and biohazardous material stored outside of the facility, is the storage device lockable?

Yes – unlock with key

How are suspicious or illicit activities reported?

Ask other staff/students if they know anything about what was seen. Authorized workers instructed to contact PI and U of S Protective Services (966-5555) to report any suspicious or illicit activities.

If a biological/biohazardous material has been reported as missing or stolen, what are the steps taken to deal with this situation? (Reference building-specific Emergency Response Plan as a resource).

If a biological material was reported missing or stolen, notify PI and Safety Resources immediately. Protective Services may need to be called if required.

Part B: Risk Threat Analysis (for Biosafety Group Use ONLY)Risk Level Analysis

FOR BIOSAFETY GROUP USE ONLY

Threat Scenario Identified Threat Level Risk Statement

6 Work Locations

Table 5 can be used to document the locations where the organism(s), biological materials and biohazardous materials will be used and stored. Table 6 can be used to provide a description of the facility and safety equipment (e.g. biosafety cabinets, autoclaves) that will be utilized.

Table 5: Biosafety Permit Work and Storage Locations.Location Type of Research1

(in vitro, in vivo, &/or both) Purpose of Work Area2 Containment Level3

Building Room #Research Annex Building

(RANNEX) 146 In vitro Research, tissue culture, lab 1 and 2

Research Annex Building (RANNEX) 146 - Hallway In vitro Storage, -20ºC Freezer in

hallway, locked 2 (freezer unit only)

W.C.V.M.. Animal Care Unit 1592 In vivoLarge Animal CL2 room,

animal husbandry, surgical procedures

2


Are visitors supervised and sign a log book, which includes name, date and time-in and time-out, purpose of visit?

Yes, visitors are supervised at all times and signed in and out in a logbook along with the purpose of the visit.

Is the intellectual property, including lab books, electronic files, etc. secured and how is this done?

Lab books stay in secured laboratories and electronic files are stored in computers which are password protected. A shared laboratory drive will be used to backup data and IP.

Who is responsible to verify and check the biological and biohazardous material inventory? How often is the inventory updated?

Dr. Legacy verifies inventory updated by the laboratory Tech, currently Jenny Liang. Inventory will be updated every 6 months or more often as needed.

List the individuals who has access to the biological and biohazardous material inventory and documentation.

- Dr. Norman Legacy (PI)- Jeff Smith (Lab Technician)- Mehan Grouse (Master’s student)- Stewart Little (Post doc.)

If the biological and biohazardous material stored outside of the facility, is the storage device lockable?

Yes – unlock with key

How are suspicious or illicit activities reported?

Ask other staff/students if they know anything about what was seen. Authorized workers instructed to contact PI and U of S Protective Services (966-5555) to report any suspicious or illicit activities.

If a biological/biohazardous material has been reported as missing or stolen, what are the steps taken to deal with this situation? (Reference building-specific Emergency Response Plan as a resource).

If a biological material was reported missing or stolen, notify PI and Safety Resources immediately. Protective Services may need to be called if required.

1 State the type of research which will be conducted at each location: in vitro or, in vivo, or both.2 State what activity will be conducted at each location: research, diagnostic, storage, autoclave, class room, surgical room, animal holding room, animal surgery room, post-mortem, biowaste storage area, etc. 3 Depending on the required containment level, there will be specific facility location, access, design and construction, air handling, containment perimeter, laboratory services, and safety equipment. Refer to the PHAC and CFIA’s Canadian Biosafety Standards (2nd Ed., 2015) and other Containment Standards.

Table 6: Safety Equipment Inventory and Location.Equipment 1 Location of Equipment

Type of equipment Model (if applicable) Serial number Building Room

Biosafety Cabinet BM4-2A 7328University # 0015984 RANNEX 146

Chemical fume hood Kem Metal University# 0065165 RANNEX 146

Flammable Safety Cabinet

Justrite™ Sure-Grip™ EX Flammable

Safety Cabinet (170 L)

2567987 RANNEX 146

Flammable Safety Cabinet

Justrite™ Sure-Grip™ EX Flammable

Safety Cabinet (47 L)

University# 0000-11-3354 RANNEX 146

Corrosives/Acids Safety Cabinet

Justrite™ Chemcor™

Lined Countertop

Corrosives/Acids Safety Cabinet

4256738 RANNEX 146

Emergency eyewash/shower Haws N/A RANNEX 146

Chemical spill kit Spill-X Chemical Spill Treatment

Kit

N/A RANNEX 146


(Part no. 78774)

Acid spill kit SpillSolv (SX1310-1) N/A RANNEX 146

Solvent spill kit SpillSolv (SX1330-1) N/S RANNEX 146

-20ºC Freezer Fridgidair 432-234 RANNEX 1461 Safety equipment includes but is not limited to, biosafety cabinets, fume hoods, autoclave, emergency eyewash, and shower equipment.


7 Standard Operating Procedures (SOPs) and Element Requirements

Part 1 - List of SOPs:

Binders containing SOP’s in RANNEX Rm 146 with SOP’s relevant to work performed and RANNEX Emergency Response Plan. Record of trained authorized workers and site specific training recordss maintained in each binder. .

1) RANNEX Local Emergency Response Plan 2) SOP-01 Operation and maintenance of a biosafety cabinet3) SOP-02 Operation and maintenance of temperature-controlled shakers4) SOP-03 Operation and maintenance of benchtop and microcentrifuges5) SOP-04 PRV and PEDV Culture from Frozen Isolates (attached)6) SOP-05 Tissue and blood collection of infected pigs (attached) 7) SOP-06 Viral Isolation from Pigs - Swabbing8) SOP-08 Plasmid Isolation from virus9) SOP-10 Preparation of competent bacterial cells (E. coli)10) SOP-11 Recombinant Protein Production using E. coli as a host11) SOP-12 Protein Purification using Gravity-Flow Affinity Chromatography12) SOP-13 Preparation and handling of SDS-containing polyacrylamide gels13) SOP-14 Preparation and handling of Safe Green (EtBr replacement) -containing agarose

gels14) SOP-15 Solid phase extraction of viral culture supernatant15) SOP-16 Liquid-liquid extraction of viral culture supernatant16) SOP-17 Operation and maintenance of rotary evaporator17) SOP-18 Operation and maintenance of chemical resistant speed vacuum18) SOP-19 Operation and maintenance of Real Time PCR and PCR Mastercycler19) SOP-20 Tissue culture of porcine and human cell lines20) SOP -21 Transport of infected pig samples between RANNEX and WCVM ACU21) SOP-22 Biowaste SOP22) SOP-23 Storage of biological materials in -20ºC Freezer23) WCVM ACU Site Specific Large Animal CL2 – Entry-Exit SOP, Animal Husbandry SOP,

Animal Room Decontamination SOP, and Animal Post Mortem Room SOP24) Animal Use Protocol # 20190005

Safety Resources2019 24 | P a g e

Part 2 - Required SOP Elements


Table 7: Summary of required standard operating procedures elements

7.1 Training Response

List the authorized workers’ and the permit holders’ roles and responsibilities. (Refer to the Biosafety Code of Practice, found at www.safetyresources.usask.ca)

- Dr. Norman Legacy (PI) (ndl123)- Jeff Smith (Lab Technician) (jrs758)- Mehan Grouse (Master’s student) (meg458)- Stewart Little (Post doc.) (swl023)

List the type of training that is required. (site-specific and Safety Resources-required training)

The training should include but is not limited to the following mandatory training for authorized workers:General (institutional):

Employee safety orientation Supervisor safety orientation (if supervising

individuals) Laboratory safety WHMIS 2015 Biosafety Biowaste online course

NOTE: Biosafety and Laboratory Safety Courses expire every 3 years.

Site specific: Site-specific safety orientation and training Operating procedures, requirements and rules

including health and safety control measures Emergency preparedness and response Other site specific procedure (e.g. tissue

culture, equipment, assays, etc.) Refresher training is required annually for Site

specific training on Standard Operating Procedures and Building or Site Specific Emergency Response Plan; Incident Reporting; and Permit Holder Biosafety Plan.

Records of required training will be kept in each SOP and Safety Training binder in Rm 146

The following required training for authorized workers:- Employee safety orientation- Laboratory safety - WHMIS 2015- Biosafety - Biowaste online course- Site specific training and ERP

All personnel must read all relevant SOPs and list these on their site specific training records which are signed and dated by the personnel and PI.

How is site-specific training delivered, verified, and documented (e.g. probationary period with supervision, quizzes, spot checks, direct supervision)?

Individuals will read the required SOPs and sign and date the training record. Then they will be instructed and/or watch the procedure being performed and then perform the procedure under supervision. The supervisor will sign and date the training record when the individual is adequately trained to perform the task themselves.

What are re-training requirements and the frequency? Site specific training (reviewing SOPs) will be required yearly. Also specified in each SOP

How are training records maintained and for how long?Records of all training are maintained by the permit holder for a minimum of 5 years. All training records kept in a blue binder in the lab above the computer station.

List all authorized workers, including their names and All authorized workers are listed on the biosafety


NSID. permit and in Section 7.1 of table 7.

7.2 Facility access and control ResponseIs there signage posted for the facility, including PPE entry requirements, emergency contact information, and containment level?

Yes

Please state the minimum PPE requirements for facility entry.

Laboratory coat, closed toe shoes, full length pants, long hair tied back

State how the PPE is donned and doffed when entering/exiting the laboratory.

Working with hazardous materials requires lab coat, gloves, and safety glasses

Lab coat, gloves and safety glasses to be removed prior to leaving the lab, wash hands with soap and water for 20 seconds.

Lab coats are all on individual labelled hooks at the entrance to the lab.

No lab coats, gloves, or safety glasses are allowed outside of the lab.

For Large Animal CL2: refer to ACU specific entry and exit procedure.

Describe the traffic flow of biological/biohazardous materials from “clean” to “dirty” areas.

All work with biohazardous materials conducted in the BSC, which is the dirtiest area in the lab; the cleanest area of the lab is the dedicated paperwork station where the computer is located; the storage of biohazardous materials is taken to the -20ºC freezer in the hallway, which is transported in a cooler, which is packed with a primary container, placed into a secondary container with absorbent material placed in between; surface decontamination conducted in the BSC in the lab with 70 % ethanol with a contact time of 5 minutes.

7.3 Hazardous material inventory, management, and waste disposal Response

State how the inventories of hazardous materials managed and how often they are updated.

Each hazardous material has an excel sheet for inventory which is updated as materials are used and/or moved. Hard copies of -20ºC inventory are available near the designated freezers.

State how often the inventory is updated. Every 6 months, and as required

Who is responsible for maintaining inventories? Technician and/or supervisor. Other lab members if required.

Where is the hazardous materials inventory kept? Computer in a shared drive and a hard copy in Rannex Rm 146.

How are the hazardous substances labelled? Warning symbols/signs are placed on all


hazardous substances. Handling procedures and disposal of hazardous waste are labelled in accordance e in accordance with the U of S Hazardous Waste Disposal Standard. Individuals who handle, package and dispose of waste must be trained.

Where and how are the hazardous materials stored? Handling procedures and disposal of hazardous waste are in accordance in accordance e in accordance with the U of S Hazardous Waste Disposal Standard and the U of S Lab Safety Manual.

Storage of hazardous substances in compatible groups, in proper containers, and with the sufficient security.

Are there any physical or operations requirements for material storage rooms/equipment (e.g. containment, security, handling, PPE)?

Yes – the -20°C freezer in hallway outside lab (Rm 146)

Freezer is locked at all times, and the key is kept in a secure location in a locked drawer, which only can be accessed by the authorized workers.

Describe the procedures that are followed if a hazardous material were to go missing or was stolen.

Personnel follow the RANNEX ERP Section XIV. Loss or Theft of Hazardous Materials.

Notify PI and the Biosafety group If missing or stolen biohazardous materials. Contact Protective Services at 306-966-5555.

What type of hazardous waste is generated? Biohazardous waste (tissue culture waste, gloves, plates, serological pipettes, etc.) and chemical waste (solvents, buffers)

Describe how hazardous waste is collected, packaged, stored, and disposed.

The PI and authorized workers ensure all hazardous waste generated is disposed of in accordance with the U of S Hazardous Waste Disposal Standard (refer to: www.safetyresources.usask.ca) .

Hazardous waste is never disposed through regular garbage or to the environment (e.g. sewer, sinks, drains).

Chemical waste must be collected in containers with screw cap lids. Each container must be clearly labelled to WHMIS 2015 standard and stored in designated area until disposed by the Waste Management, Safety Resources.

SHARPs (needles, razor blades, glass slides, etc.)


http://www.safetyresources.usask.ca/

are placed into a commercially available SHARPs container and disposed as biohazardous waste.

Liquid biohazardous waste inactivated prior to disposal with a fresh bleach solution of min. 1% v/v with a contact time of 20 minutes prior to disposal down the sink.

Solid Biohazardous waste must be collected in a biohazard bag in the designated container by the BSC, then tied, surface decontaminated with 70% ethanol with contact time for 5 minutes prior to removal from BSC< and then placed into the lined grey biohazard waste bin.

Solid biohazardous waste (non-anatomical) disposed through Waste Management (using lined grey bins with bar codes). Grey bins obtained from biowaste shed. Key obtained from department office. Follow the U of S Biowaste guide. All individuals must take the online biowaste course prior to using the grey bins.

Infectious animal carcasses are placed into incinerate pails at ACU and sent through Waste Management for incineration.

Health animal carcasses are placed into a carrying tote, and transported to PDS for disposal in accordance to SOP 24 – Disposal and Transport of animal

Biohazardous materials stored in a “dirty’ fridge in a secondary container (bag or bin) or in cryovials at -20oC.

7.4 Biohazardous material procurement and transport Response

Describe how the lab procures biohazardous materials. Refer to the U of S Procurement and Transfer ofOrganisms and Biological Materials.

Permit holders who wish to procure biological materials must notify the Biosafety group prior to procurement.

The permit holder must ensure the biological material is listed on the biosafety permit. If the biological material is not listed, a permit amendment is required. For risk group 2 or 3 biohazardous materials, the


permit holder must complete the Biohazardous Agent Transfer Notification Form (www.safetyresources.usask.ca) and submit the completed form to the Biosafety group for review and approval. The Biosafety group will then determine if any other import permits and/or certifications are required for procurement of the biological material (e.g. an import permit and compliance checklist from CFIA).

An material transfer agreement (MTA) may be required and the permit holder will contact the Innovation Enterprise office.

Describe how materials are moved with the facility, between facilities, or off campus.

Materials are moved within the lab (containment zone) using practices described in Section 20.1.1 of the Canadian Biosafety Handbook. All movement is using a primary leak proof, impact resistance container placed into a secondary leak proof, impact resistance container with absorbent material. A surface decontamination of the secondary material required with 70% ethanol with a contact time of 5 minutes.

If required to move outside of the containment zone, place secondary container into a third carrying container such as a Styrofoam cooler and carried to the destination.

Refer to SOP -21 Transport of infected pig samples between RANNEX and WCVM ACU.

Describe the PPE that is required to transport material. In the lab: Lab coat, gloves, safety glasses, closed toe heeled shoes, hair tied up

Between buildings: remove lab coat prior to leaving lab, pants, closed toe heeled shoes

State who is notified if the materials are to be moved within the facility, between facilities, or off campus.

RG 2 and 3 biohazardous materials may only be transferred to another permit holder at the U of S with appropriate approvals by the Biosafety group and BPAC.

Describe who is notified if the materials go missing or are stolen during the transfer.

Personnel follow the RANNEX ERP Section XIV. Loss or theft of hazardous materials. Contact the permit holder and Safety Resources.

7.5 Specific procedures and techniques related to the nature of research

Response

What are the specific laboratory tasks, procedures, and techniques being performed under this permit?

Gel electrophoresis, tissues culture, viral propagation, RT PCR, transfection, immunohistochemistry, bacterial culture, sub cloning, animal studies with pigs



For more details, refer to the SOPs listed in section 7.

Who is performing these tasks? Permit holders and authorized workers

Who is responsible for developing and approving procedures?

Permit holder is responsible for developing and approving procedures.

Where are SOP’s related to operating procedures stored (e.g. written procedures, books, papers, worker experience, verbal communication)?

SOPs (written) are located in a binder labelled SOPs on the bookshelf above the lab computer workstation. Electronic versions found on lab computer desktop in folder Lab SOPs.

Do procedures include appropriate health and safety requirements (e.g. controls, PPE)?

Yes

How are staff/students made aware of and trained on procedures?

All authorized workers must read the SOPs prior to commencing work in the lab. This is then verified by the PI and the trainees and PI sign the Site Specific Training record. All personnel are also visually monitored by the PI for effective practice of SOPs.

Re-training od SOPs are reviewed annually and is documented.

How often are the specific laboratory procedures reviewed and updated?

Annually and as required.

7.6 Immunization and medical surveillance (if applicable)

Response

Is a medical assessment required prior to an individual beginning work (e.g. identifying known allergies, prior immunizations, immune status, etc.)?

If required, complete the Medical Risk Assessment Form.

Not at this time. Working with Terrestrial animal pathogens.

How is the individual informed of all safety mitigation measures against working with the pathogen and toxin?

The following required training for authorized workers are required:- Employee safety orientation- Supervisor safety orientation- Lab safety (online)- WHMIS 2015 (online)- Biosafety (online)- Biowaste (online)

Is there an on-going medical surveillance program? Describe.

None is required at this time.

If applicable, how is the medical surveillance program monitored and tracked?

n/a

Are any vaccinations required? No vaccinations required at this time.

7.7 Working alone or after hours ResponseWhat work is permitted to be carried out alone or after Supervisors are responsible for scheduling of work


hours? done outside of hours. Supervisors are to review tasks with the employee who is performing the duties outside of the normal workday. Staff members are only to perform the tasks requested by their supervisor during these hours. PI encourages workers to work in pairs whenever possible.

No animal work to take place outside of business hours unless approved by PI.

How are working alone activities monitored and tracked?

When working alone, staff will have access to a phone (cell phone) so that they are able to contact emergency services immediately. Supervisors check on their staff either by phone or by person during the scheduled work period.

Staff are provided with a contact number for their supervisor in case of an emergency.

What procedures are in place to protect individuals working alone (e.g. restricted activities, communications, emergency preparedness and response)?

Protective Services sticker will be affixed to internal lab and office telephones. Employees will be encouraged to use the “Safewalk” program so that they can be escorted for evening walks on the campus grounds.

Contact numbers include:

Dr. Legacy cell: 306-969-7777Protective Services 5555

7.8 Facility and equipment care and maintenance

Response

State how the facility maintenance concerns, needs, and requirements are managed.

All concerns and needs are discussed with the PI. The PI will contact Customer Service Centre to have repairs and/or maintenance needed. Conduct bi-weekly meetings with authorized workers to discuss any items.

What are the housekeeping requirements for the facility?

All authorized workers are instructed and monitored for keeping their bench/work spaces free of clutter and clean.

What are staff/student responsibilities for housekeeping and for facility/equipment maintenance and care?

Custodial staff are allowed to change the soap dispenser, garbage, paper towels, and clean floor (under direction of the PI or designate)

Authorized workers are tasked with cleaning equipment, spill clean up, counter-top clean-up, glassware clean-up, waste disposal, etc.

Authorized workers conduct daily site checks and site specific inspection is contacted by personnel every 3 months and documented.


Equipment is regularly inspected by all personnel and if requiring cleaning or maintenance, refer to equipment manual. For equipment repairs, consult with PI.

Under what conditions is laboratory equipment to be operated (e.g. procedures, PPE, cleaning, decontamination)?

Authorized workers refer to SOPs which detail operating, cleaning, decontaminating procedures for individual lab equipment.

What are the maintenance and certification requirements for the equipment?

Authorized workers are instructed to the manufacturer’s operation manual stored in the binder above the computer work station in the lab.

BSC are certified annually by third party service provider (e.g. Con-Test). Once certified, a certification report is provided and a certification sticker is posted on the BSC. Certification expires 1 year from the date it was tested.

All fume hoods are also inspected yearly and posted on the units (certification done by Safety Resources). Certification expires 1 year from the date it was tested.

What records are required operationally for the equipment?

The BSC is inspected yearly and certifications posted on the unit. Certification reports are kept on record in the equipment binder found above the computer work station.

Each piece of equipment has an equipment use log, which records the use, cleaning, repair/maintenance of it. Calibration certificates are documented as well in the equipment binder.

7.9 Disinfection and decontamination ResponseWhat disinfectants are used in the lab? State concentration, whether the solution is to be made fresh, and contact time?

Main disinfectant for surface decontamination: - 70% ethanol with 5-minute contact time; Mix

date of ethanol solution to be placed on spray bottle.

- Using hydrogen peroxide wipes (contact time of 5 minutes) for surface decontamination of electrical equipment and secondary containers when transporting.

Biohazardous Liquid Waste:- Use a 10% bleach solution made fresh with a

contact time of 20 min (1 part bleach, 9 parts


liquid waste) (e.g. for vacuum pump apparatus)Which disinfectants are used for each biological/biohazardous material?

For virus work and cell culture: - 70% ethanol and 10% bleach for liquid waste

Main disinfectant for surface decontamination: - 70% ethanol with 5-minute contact time; Mix

date of ethanol solution to be placed on spray bottle.

- Using hydrogen peroxide wipes (contact time of 5 minutes) for surface decontamination of electrical equipment and secondary containers when transporting.

Biohazardous Liquid Waste:- Use a 10% bleach solution made fresh with a

contact time of 20 min (1 part bleach, 9 parts liquid waste) (e.g. for vacuum pump apparatus)

How is the biohazardous waste sterilized or inactivated?

Biohazardous waste:

Solid – sent away for autoclaving through Waste Management

Liquid: using freshly made 10% bleach solutionAre there specific disinfection processes to be followed (e.g. for specific hazardous materials, laboratory surfaces, equipment, spills)?

Any repair, maintenance, or relocation of the biosafety cabinet requires a gas decontamination by a third party service provider, Con-Test. Contact the service provider to arrange to have this done: Leighton Davey 306-251-0185, [email protected]

7.10 Workplace monitoring ResponseHow are facilities and work activities monitored? Monitored visually by the PI. All personnel self-

monitor and monitor colleagues. Who is responsible for monitoring the facility and faculty, staff and students working in the facility?

PI

How frequent are workplace monitoring activities conducted?

Daily

How are workplace monitoring activities documented? (e.g. self-inspections, meetings)?

Self-inspections are carried out every 3 months and documented in a binder in the lab above the computer workstation.

How are safety issues and incidents managed? Safety issues and incidents are managed according to guidelines in the Laboratory Safety Manual Section 9.2 (www.safetyresources.usask.ca)

How are safety matters communicated with faculty/staff/students?

Safety matters are communicated directly to personnel when warranted and covered in biweekly lab meetings when needed.

7.11 Animal work considerations (if Response



applicable) What are the specific procedures carried out when working with animals (e.g. entry/exit procedures, training requirements, restraints and handling of the animals, decontamination, animal transport, etc.)?

Any authorized workers using animals would be required to take the Animal Ethics courses (online) e.g. basic, lab animal and large animal handling. Refer to AUP 200190005.

What health and safety requirements are implemented when working with animals (e.g. engineering controls, procedures, PPE)?

Authorized workers follow all PPE requirements stated in the AUP SOPs entry and exit for large animal CL2. PPE is dedicated to the facility.

7.11 Human work considerations (if applicable) (Includes clinical samples,

phlebotomy)

Response

What are the specific procedures carried out when working with human blood, cells, tissues, or bodily fluids (e.g. PPE requirements, training requirements, decontamination, etc.)

- Cell culture and viral propagation as outlined in SOP-20 Tissue culture of porcine and human cell lines

What health and safety requirements are implemented when collecting human blood, tissue, or bodily fluid samples (e.g. engineering controls, procedures, PPE)?

- Use CL2 operational processes outlined in SOP-20 Tissue culture of porcine and human cell lines

7.12 Emergency preparedness and response

Response

State which Emergency Response Plan (ERP) the lab follows for emergency procedures.

RANNEX ERP Feb 2016

Where is the ERP located? On the desktop of the lab computer and in a binder above the computer workstation labelled ERP.

Describe situations that may occur in the lab that need to be captured in the ERP.

Situations that could occur in the lab and covered in the ERP: Fire, spills of hazardous materials, and medical emergencies.

State how the annual ERP training is provided and documented.

ERP found on the desktop of the lab computer and in a binder above the computer workstation labelled ERP.

ERP review is conducted annually at lab meetings and training is documented. Reminders are provided periodically in lab meetings.

8 References


1. Canadian Biosafety Standard and Canadian Biosafety Handbook, 2015, 2nd Ed. https://www.canada.ca/en/services/health/biosafety-biosecurity.html

2. Safety Resources website: www.safetyresources.usask.ca 3. McFall-Ngai, M., Hadfield, M.G., Bosch, T.C., Carey, H.V., Domazet-Loso, T., Douglas,

A.E., Dubilier, N., Eberl, G., Fukami, T., Gilvert, S.F., et al. (2013). Animals in a viral world, a new imperative for the life sciences. Proc Natl Acad Sci USA 110, 3229-3236.

4. Tettelin, H., Masignani, V., Cieslewicz, M.J., Donati, C., Medini, D., Ward, N.L., Angiuoli, S.V., Crabtree, J., Jones, A.L., Durkin, A.S., et al. (2005). Genome analysis of multiple pathogenic isolates of PRV. Proc Natl Acad Sci USA 102, 13950-13955.

5. Ruzzini, A.C., and Clardy, J. (2016). Gene Flow and Molecular Innovation in PRV. Curr Biol 26, R859-864.

6. Ochman, h>, and Moran, N.A. (2001). Genes lost and genes found: evolution of viral pathogenesis and symbiosis - PDV. Science 292, 1096-1099.

7. Jain, R., Rivera, M.C., Moore, J.E., and Lake, J.A. (2003). Horizontal gene transfer accelerates genome innovation and evolution. Mol Biol Evol 20, 1598-1602.

8. Cordero, O.X., and Polz, M.F. (2014). Explaining microbial genomic diversity in light of evolutionary ecology. Nat Rev Microbiol 12, 263-273.

9. Polz, M.F., Alm, E.J., and Hanage, W.P. (2013). Horizontal gene transfer and the evolution of PRV. Trends Genet 29, 170-175.

10. Jorgensen, T.S., Kiil, A.S., Hansen, M.A., Sorensen, S.J., and Hansen, L.H. (2014). Current strategies for mobilome research. Front Microbiol 5, 750.

11. Sentchilo, V., Mayer, A.P., Guy, L., Miyazaki, R., Tringe, S.G., Barry, K., et al. (2003). Community-wide plasmid gene mobilization and selection. ISME J. 7, 1173-1186.

12. Jones, B.V., Marchesi, J.R. (2007). Transponson-aided capture (TRACA) of plasmids resident in the human gut mobile metagenome. Nat. Methods, 4(1), 55-61.

13. Schwyn, B., Neilands, J.B. (1987). Universal chemical assay for the detection and determination of siderophores. Anal. Biochem. 160(1), 47-56.

14. Donovan, C.J., Garduño, R.A., Kalmokoff, M.L., Ku, J.C., Quilliam, M.A., and Gill, T.A. (2009). Pseudoalteromonas Bacteria Are Capable of Degrading Paralytic Shellfish Toxins. Applied and Envi. Micro., 75(21), 6919-6923. doi : 10.1128/AEM.01384-09

15. https://www.atcc.org/products/all/15057.aspx 16. https://www.atcc.org/Products/All/51841.aspx 17. Virusdisease. 2019 Jun;30(2):294-301. doi: 10.1007/s13337-019-00513-w. Epub 2019

Feb 9. Isolation and recombinant analysis of variants of porcine epidemic diarrhea virus strains from Beijing, China. Wang P1, Zhu J1, Liu X1, Guo J1, Gu X1, Ruan W

18. Virology. 2018 Apr;517:188-198. doi: 10.1016/j.virol.2017.11.024. Epub 2017 Dec 14.Differential gene modulation of pattern-recognition receptor TLR and RIG-I-like and downstream mediators on intestinal mucosa of pigs infected with PEDV non S-INDEL and PEDV S-INDEL strains. Temeeyasen G1, Sinha A2, Gimenez-Lirola LG3, Zhang JQ2, Piñeyro PE4.


https://www.atcc.org/Products/All/51841.aspx

https://www.atcc.org/products/all/15057.aspx


https://www.canada.ca/en/services/health/biosafety-biosecurity.html

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