Upload
others
View
9
Download
0
Embed Size (px)
Citation preview
2017
Nagasaki UniversityNagasaki UniversityEducation lectureEducation lecture
General course
Committee for biosafetyCommittee for biosafety生物災害等防止安全運営委員会
1
Contents1.Rules and regulations for possession of pathogens
2 Properties of pathogens and Safety management2.Properties of pathogens and Safety management(Biosafety and Biosecurity)
3 Safety handling for pathogens3.Safety handling for pathogens
4.Basic operation in laboratory (BSL-2)
5.Other regulations
Rules of Biosafety management for Nagasaki University1.Properties of pathogens and Safety management2.Safety handling for pathogens according to BSL2.Safety handling for pathogens according to BSL3.BSL for experiments planed4.Measures against incidents and emergencies5.Physical and biological containmentsy g6.Safety management for experimental procedures7.Other concerns for biosafety
2
1. Rules and regulations for possession of pathogens
3
Rules and regulations for usage and possession of pathogens
Rules of Biosafety management for Nagasaki UniversityInstruction of Biosafety management for pathogen researchI.
II. Act on the Prevention of Infectious Diseases and Medical Care for Patients withInfectious Diseases (Japanese law: Kansenshouhou)
y g p g
Specified pathogens:Class I, II, III and IV PathogensInfectious Diseases (Japanese law: Kansenshouhou)
(The term "Specified Pathogens, etc." as used in this Act means Class I Pathogens, etc., Class II
III Domestic Animal Infectious Diseases Control Law
Pathogens, etc., Class III Pathogens, etc., and Class IV Pathogens, etc.)
III. Domestic Animal Infectious Diseases Control LawMonitoring infectious disease pathogens: Emphasis management Domestic Animal Infectiouspathogens, Management Domestic Animal Infectious pathogens, Notifiable diseases such as pathogens
・Act on the Conservation and Sustainable Use of Biological Diversitythrough Regulations on the Use of Living Modified OrganismsIV.
Organisms of class 2, 3, and 4
Regurations for animal experiments, radiation management, and so on 4
Check the BSL of the pathogens
Specified pathogensMonitoring infectious disease pathogens
Registration process
1. 2. 3.
BSL1A microorganism that is unlikely to cause human or animal disease. BCG vaccine strain,
p g disease pathogens
BSL2
attenuated live vaccine and Adeno-associated virus
Class IV pathogens, etcClass III pathogens, etcClass II pathogens, etc
Protozoa, Trematoda, Taneia, Caenorhabditis, Fungi such as A.fumigatus etc, Bacteria such as H.pylori, M.avium etc, Virus such as measles virus
N ifi bl VSV tDengue virus, Shiga toxin, C.Parvum (genotype I and II)etc
Monkeypox virus, Rabies virus (fixed virus)
C.botulinum,Botulinum toxinetcManagement
Notifiable VSV etc
SFTS virus,R.Japonica etc
West Nile virus,Serovar Typhi etc
BSL3SARS virus,B.Anthracis etcEmphasis management
Management
Fungi such as B.dermatitidis etc, Bacteria such as M.Africanum etc, Virus such as Chikungunya virus etc
Emphasis management
Class I pathogens, etcLassa virus, Zaire virus, Crimean-Congo Heamorrahgic virus etc
(BSL4)
5
I.Registraion for Biosafety management for pathogen research
6
I. Registration for Biosafety management for pathogen research
BSL1,2➡・Notification for usage and storageEnd report・End report
BSL3 ➡・Request for usaged tand storage
・Request for supply・End report
Specified pathogens, Monitoring infectious disease pathogens➡Registration process for specified➡Registration process for specifiedpathogens but not
7
I. Registration for Biosafety management for pathogen research
8
I. Registration for Biosafety management for pathogen research
9
II. Registration for Rules of Biosafety management for Nagasaki University(Specified pathogens)
10
11
Class I Class II Class III Class IV
Duty and penalty of a holder of specified pathogens
P i i
Designation by the Minister of Health, Labourand Welfare
Class I Class II Class III Class IV
Permission
NotificationPreparation of Infectious diseases prevention
Education and training
Assignment of supervisor
rule
Education and training
Record of usage and storage
Sterilization etc (in case of rescission of permission)
Record of usage and storage
Standard of facility
Standard of storage
Notification of transfer
Accident report
g
Coping with disaster
Duty and penalty Order to improvement12
II. Registration for Rules of Biosafety management for Nagasaki University(Specified pathogens)
RegistrationClass II, III and IV pathogens, etc➡ Request for the facility,
End report for the facilityEnd report for the facility,Request for usage,Request for sterilization,Request for supply,Request for supply,Request for transport
Registration and notification to Ministry of Health, Labour
d W lfand Welfare
13
Request for usageRequest for the facility
14
Request for supply Request for transport
15
III. Registration for Domestic Animal Infectious Diseases Control Law
16
III. Registration for Domestic Animal Infectious Diseases Control Law
Registration
Emphasis management Domestic Animal Infectious pathogens,Management Domestic AnimalManagement Domestic Animal Infectious pathogens,Notifiable diseases such as pathogens ➡Request for the facility,Request for the facility,
End report for the facility,Request for usage,Request for sterilization,q ,Request for supply,Request for transport
Registration and notification toMinistry of Agriculture, Forestry
d Fi h iand Fisheries
17
Nagasaki UniversityOrganization for storage and usage of pathogens in Nagasaki University
・Pathogens Supervisor
Nagasaki University
・Holder of specified pathoigens (Dean)
・Safety officer
Pathogens SupervisorDepartment
Safety officerLaboratory for Specified pathogens and Monitoring infectious disease pathogens
Laboratory for pathogens
・Researcher (lab member)・Responsible party
infectious disease pathogens
R h (l b b )・Responsible party
・Researcher (lab member)
P ll d t t・Researcher (lab member)
・Researcher (lab member)・Researcher (lab member)・Person allowed to enter
・Person allowed to enter・Person allowed to enter
・Person allowed to enterPerson allowed to enter
18
Responsibility of Responsible party
・Registration and notification for usage storage and
・Preparation of manuals and standard operation
・Registration and notification for usage, storage and transfer of pathogens Experimental
planp p
procedures (SOP)
Education,・Practical education for operations in each laboratory・Practical training in each laboratory・Education for a person to laboratory temporarily
training
Facility,E i t
・Record of pathogen usage, storage and transfer・Maintenance for safety equipment
record
Equipment
・Preservation of records for five years・Record of entrance and exit
M f i id t d i
record
Incidents, ・Measures for incidents and emergencies・Others according to Rules of Biosafety management for Nagasaki University,Instruction of Biosafety management for pathogen research
emergencies
Safety officer
y g p g
19
2.Properties of pathogens and Safety management
20
PathogensParasites BacteriaParasites Bacteria
Toxin
FungiVirus
Toxin
Prion
・病原体等(長崎大学生物災害等防止安全管理規則)病原微生物及び動植物が産出する毒性物質等,生物学的相互作用を通して人体や環境に災害を及ぼす可能性のある物質
・特定病原体等、監視伝染病病原体(長崎大学生物災害等防止安全管理規則)病原体等のうち 感染症法 家畜伝染病予防法施行規則に規定する病原体
・病原微生物(研究用微生物安全管理要領)哺乳動物等に対して病原性を持つ微生物(原核生物、真菌、ウイルス、ウイロイド、原虫、寄生虫及びプリオン)
病原体等のうち、感染症法、家畜伝染病予防法施行規則に規定する病原体
21
Route of infection
Droplet infectionDroplet nuclei infection (aerosol transmission)
Contact infectionPhysical contact with the person suffering the disease
Respiratory infection
Droplet nuclei infection (aerosol transmission)gCasual contact with their secretions or objects touched by them or airborne route among other routes
Food, water etcM i i k
Oral infectionVector-borne infection
Mosquito, tick, etc
Perinatal Blood-borne infection
Vertical infection
through contamination by
Conditions of infection
through contamination by blood and other body fluids
・Route of infection・Immune status (sensitivity, resistance)
Conditions of infection・Types of pathogens ・Infectious dose (quantities)
22
Biosafety and Biosecurity
Bi h d Bi t i
Bio-riskBiohazard Bioterrorism
A threat to the health of humans and other living organisms
Intentional release or dissemination of biologicaland other living organisms dissemination of biological agents
Risk control
Biosafety Biosecurity
Risk control
Biosafety BiosecurityPrevention humans and environments
t f th
Prevention pathogens from hetc from pathogens humans
23
Examples of Bio-risk
・Handling pathogens not using safety cabinet
・Inadequate wearing PPE
・Careless administration of controlled area
・Careless administration of pathogens
Handling pathogens not using safety cabinet
・Mistake and inadequate handling of pathogens ・Inadequate storage and handling of pathogens
Careless administration of controlled area
・Spills of pathogens from broken container
・Wrong use of syringe needle, pipette ・Malicious user
・Unauthorized experiments
BioterrorismBiohazard
・Needle-stick injury, bite
・Contact infection through a wound
・Theft and loss of pathogens
・Discrepancy of pathogens inventory
・contact and droplet infection through a mucosa
・Inhalation of contaminated aerosol
・Invasion of suspicious person
・Dual use
・Frowing out of contaminant ・Wrongful action of researchers
24
Risk-control
Containment is crucial for risk-control.
Biosafety laboratoryEntrance f b ildi
yPrevention of infection in workers,Prevention of flowing out from laboratory
Bi it
Freezer etcSafety cabinet
of building
E t f l bTube etcBiosecurityControl of access to pathogens Controlled
area
Entrance of labTube etc
Hardware and softwarehardwareBuilding equipment
Before and after measure
beforePreliminary measuresBuilding, equipment, Preliminary measures
afterPreparation of measures for restoration
softwareAdministration of safety and security
According to pathogens
p
According to pathogens
Biosafety level (BSL)-1, BSL-2, BSL-3, BSL-425
Biosafety Biosecurity
Examples of bio-risk control
Set up of safety cabinet
Set up of safety equipment
Locking mechanism for the building
Designation of a Locking mechanism for the laboratory
Isolator for animal husbandry Locking device for storage freezer
Designation of a controlled area
g y
Air-conditioning unit with HEPA filterAccess control system for
hard
wa
Sterilization equipment
Display of evacuation route
laboratory
Record of laboratory entry and exit
are
Wearing PPE
Record of inventory for pathogensUse safety cabinet!
Periodic inspection of inventory
Training
Adequate disinfection and sterilization
Restriction of sharpsCorrect knowledge and act of pathogen research
Manuals
softw
are
Usage of safety equipment
Health care for workers SOPs
Adequate disinfection and sterilization
Report of theft and loss
Preparation for emergency
Appropriate coping with accidents
Correct knowledge and act of dual use
Measures are according to biosafety levels26
3.Safety handling for pathogens
27
Containment of laboratory according to biosafety level
BSL 1BSL-2
Safety cabinetA tocla eBSL-1 Autoclave
BSL-3 BSL-4(suit type)HEPA filter
HEPA filter
l b t
Changing roomPersonal shower Suit room
Chemical shower
anteroom
HEPA filter
laboratoryAir Air laboratoryAir flow
Air flow
Keep negative pressure in the roomKeep negative pressure in the room
28
Confirm the standards of structure and equipment according to biosafety level in Rules of Biosafety management for Nagasaki University
29
Examples of BSL-2 laboratory
30
Personal Protective Equipment (PPE)
BSL-2 BSL-3
Surgical maskFace guard
N95 k
Gloves
N95 mask
Overall, apron
Lab coat Double gloves
Boots (shoe covers)
31
Safety cabinet(primary containment)
Class II(a, b, c) Uase a class II type safety cabinet for pathogens!!
Class I Class III Clean bench
Don’t use for pathogen or any clinical specimen!
32
Safety equipment
Centrifuge machines
Isolator for animal housing PipetmanPipetman, pipet aid
33
DisinfectantA ti i bi l t t d t i i th t li i th bj tAntimicrobial agents to destroy microorganisms that are living on the objects.Disinfection does not necessarily kill all microorganisms.
Extreme physical and/or chemical process
Liquid chemicals Classification of Chemical Disinfection
p y p
Disninfectant Bacteria Tuberculosis
Spore Fungi Non-enveloped virus
Enveloped virus
High-level disinfectionComplete elimination of all
Glutaral + + + + + +
Peracetic acid + + + + + +microorganisms in or on an instrument, except for small numbers of bacterial spores.
Peracetic acid + + + + + +
Phtaral + + + + + +
Intermediate-level disinfection
Sodium hypochlorite + + + + ± +disinfectionElimination of most vegetative bacteria, some fungi, and some viruses in a practical period of time (<10 minutes)
Alcohols + + - + ± +
Povidone iodine + + - + ± +
Cresol soap + + - ± ± +
Low-level disinfectionElimination of most vegetative bacteria, some fungi, and some viruses in a practical
i d f ti ( 10 i t )
Quaternary Ammonium Compounds + - - ± - +
Chlorhexidine + - - ± - +period of time (<10 minutes) Amphoteric surface
active agents + ± - ± - +
+:Effect -:No effect ±:require high concentration and/or prolonged time34
SterilizationProcess that eliminates, removes, kills, or deactivates
Autoclave Dry heat
Process that eliminates, removes, kills, or deactivates all forms of life and other biological agents
Chemical indicator
Gas
・121℃, longer than 15 min
・Confirm that sterilization process successfully done by indicator・steam ➡ effective on the surface ➡ need validation for animal carcasses
35
Storage of pathogens
・Store in storage container such as freezers etc with locking device. Containers have to be registered.
・Control the area equipped storage containers.
・For stock of pathogens in freeze, use cryotubes but not the regular tubes.
・Lock the door of freezer.
・Manage the key by responsible party
・ Record inventory of pathogens
D ’t i di t th th th t・Don’t indicate the pathogen names on the storage containers
36
RecordExample of specified class II pathogenExample of specified class II pathogen
・Transfer and Receipt of pathogens(the types and quantities if they are Toxins )・Year month and date of Transfer and Receipt of pathogens・Year, month and date of Transfer and Receipt of pathogens・Means and places of storage・Type of pathogens for sterilization etc・Year, month and, means and place for sterilization of pathogens andcontaminants
・Personal full name who transferred and received pathogensPersonal full name who transferred and received pathogens・Personal full name who entered in and exited from laboratory・Year, month and of enter and exit・Personal full name who used pathogens・Personal full name who sterilized・Year month and date of inspection of facility and equipment for specified class・Year, month and date of inspection of facility and equipment for specified class
II pathogen, results of inspections and the contents of treatment, personal fullname who inspected and treatedY th d d t f d ti d t i i f th i d h・ Year, month and date of education and training for authorized persons who usespecified class II pathogen, the contents and full names of trainees
37
1. Basic Responsibilities and attitude of ScientistsScientists shall recognize that they are responsible for assuring the quality of the specialized
Code of conduct for scientists
Scientists shall recognize that they are responsible for assuring the quality of the specialized knowledge and skills that they themselves create, and for using their expert knowledge, skills and experience to contribute to the health and welfare of humankind, the safety and security of society and the sustainability of the global environment. Scientists shall always make judgments and act with honesty and integrity endeavoring to maintain and improve their own expense abilities and skills andhonesty and integrity, endeavoring to maintain and improve their own expense, abilities and skills, and shall make the utmost effort to scientifically and objectively demonstrate the accuracy and validity of the knowledge they create through scientific research.
2 Attitude of Scientists2. Attitude of ScientistsScientists shall always make judgments and act with honesty and integrity, endeavoring to maintain
and improve their own expense, abilities and skills, and shall make the utmost effort to scientifically and objectively demonstrate the accuracy and validity of the knowledge they create through scientific research Scientists shall recognize that scientific autonomy is upheld by public trust and the mandateresearch. Scientists shall recognize that scientific autonomy is upheld by public trust and the mandate of the people, understand the relationships between science, technology, society, and the natural environment from a wide-ranging perspective, and act in an appropriate manner.
3 Accountability and Disclosure3. Accountability and DisclosureScientists shall strive to disclose and actively explain the roles and significance of their own research,
evaluate the possible effects of their research on people, society and the environment as well as the changes that their research might engender, neutrally and objectively disclose the results of this evaluation, and build a constructive dialogue with society.
4. Dual Use of Scientific Research OutcomesScientists shall recognize that there exist possibilities that their research results, contrary to their
own intentions, may be used for destructive actions, and shall select appropriate means and methods as allowed by society in conducting research and publicizing the results.
Science Council of Japan January 201338
Emergency response
Prepare manuals and measures for incidents and accidents such as disaster, occurrence of abnormity of an electric facility, illness of workers Invasion of suspicious person
Prepare and indicate an emergency contact list in the laboratory
workers, Invasion of suspicious person.
If you have an accident during pathogen experiment,Don’t panicDon’t panicStop experimentsI f dInform the staff immediatelyInform everyone around
Evacuate immediately if necessary
I f l th k d fi t i tIn case of a large earthquake and fire, stop experiments, secure the pathogens, and evacuate
39
Transport of pathogensTransport of infectious and potentially infectious materials is subject to strict national
Laboratory personnel must ship infectious substances according to applicable
Transport of infectious and potentially infectious materials is subject to strict national and international regulations. These regulations describe the proper use of packaging materials, as well as other shipping requirements.
Laboratory personnel must ship infectious substances according to applicabletransport regulations. Compliance with the rules will:1. Reduce the likelihood that packages will be damaged and leak, and thereby2. Reduce the exposures resulting in possible infections3 I th ffi i f k d li
container
3.Improve the efficiency of package delivery.
Container for Category A or Category B pathogens
pathogen(primary container)
lid
seal Dry-ice Label indicating the contents
absorber
Shock absorber seal
Seconday container
container made of styrofoam
Don’t put in dry-ice into this box!!40
Responsible party should create rules a set of rules for laboratories.Practice proper biosafety and biosecurity.
Prepare manuals and SOPs, train researches and other staffs.41
Controlled area
・Designate a controlled area including laboratory andstock rooms for pathogens, room for sterilizationprocess, air-conditioning machine room and so on.
・Only authorized persons can enter in the controlled area.
・The international biohazard warning symbol and sign mustbe displayed on the doors of the rooms (controlled area)p y ( )
1. BSL2. Name and phone number of responsible party3. The international biohazard warning symbol3. The international biohazard warning symbol
Don’t indicate the type of pathogens and pathogen names!!
42
Inspection of safety equipment
Annually inspection is required for the safety equipment to use specified pathogens and Monitoring infectious disease pathogens
・BSL-3 facility
・safety cabinet
・Autoclave
・Storage freezer and refrigeratorg g
Centrifuge machine etc・Other equipment if neccessary
Centrifuge machine etc
Should be inspected by responsible party and confirmed by safetyShould be inspected by responsible party, and confirmed by safety officer
43
4. Basic operation in laboratory (BSL-2)
44
Only authorized person can enterOnly authorized person can enter BSL-2 room. Administrate with key or ID card system etc.
Change your shoes to slippers.If hIf you use your own shoes, prepare one which completely covers your toes.
Don’t open the door for a long time. Security guard comes if door remainsSecurity guard comes if door remains opened for more than 90s.
Don’t escort non‐authorized persons without permission
Don’t open the window during experimentsDon t open the window during experimentsDon’t keep opend the door。
45
Wash you hands before doing experiment.
Wear white lab coat when working with infectiousworking with infectious pathogens.
Prepare your own lab coat for BSL-2 room use only.D ’t l b t t idDon’t wear lab coat outside laboratory
46
Wear disposable gloves when using infectious pathogens.
T k ff lTake off gloves outside laboratory
When wearing gloves, don’t touch door knob and PC.
47
Book or record your experiments in the sheet (name, pathogen, time) on top of the safety cabinet
Confirm to turn on the light, fan and
Look for the waste bottle If it’s full
gas burner.
Look for the waste bottle. If it s full, autoclave it. Get another bottle for use.
Look for the a tocla ed steel can If it’sLook for the autoclaved steel can. If it’s full, autoclave it and get another one.
48
Wait 15 min after turn onWait 15 min after turn on the cabinet fan to stabilize the air flow
Place minimum number ofPlace minimum number of equipment in the cabinet.
Don’t leave any stuff on the air inlet
K l ki
air inlet.
Keep a large working space for easy handling.
49
Spray ethanol on your glove, p y y g ,when you touch something outside of the cabinet.
For spinning 15 or 50 ml tube, b k tuse bucket.
You can open the bucket only in a safety cabinet.
50
Prepare disinfectant solutions such as 70% ethanolPrepare disinfectant solutions such as 70% ethanol besides on the safety cabinet.
If ill i f ti t b t lIf you spill infectious agents, cover by paper towel, pour disinfectant gentry, place longer than 10 min, and retrieve paper towel, and then wipe them off with cotton or tissuepaper towel, and then wipe them off with cotton or tissue paper.
51
After use, take out waste chips, tubes and pipettes Put them in the steel can and thenpipettes. Put them in the steel can and then autoclave.
For pipettes, use double autoclave bags. When full take the inside bag and replaceWhen full, take the inside bag and replace with a new bag.
Discard used gloves in the steel can and then autoclave.
If autoclave machine is busy, let the can remain near the safety cabinet and autoclave later.
52
After use spray 70% ethanol toAfter use, spray 70% ethanol to equipment and wipe with cotton or tissue paper.
Rinse tubes of aspirator withRinse tubes of aspirator with disinfectant
Keep working cabinet fan 10 min after work by turn off
Keep used equipment and other
y
laboratory items under UV for at least 10 min, and then take out from the safety cabinet.y
53
Keep micropipettes and tubes in the small movable cabinet. Don’t keep inside the safet cabinetDon’t keep inside the safety cabinet.
Plug in pipette aid. Keep minimum stuff inside safety cabinet.
54
Discard used ice in a sink and place ice container under a sink.
D ’t f t t l thDon’t forget to close the stop cock of gas.
Return tube racks to the assigned shelf.
55
Confirm the correct water level of autoclave machine before use.
Program is already set at 121°CProgram is already set at 121 C, 20min. Lock the door and push start button.
56
Check indicator tape and discard autoclaved stuffdiscard autoclaved stuff separately as burnable, plastic, or aluminum foil.
Put used clinical items such as blood needles and hematocritblood, needles and hematocrit tubes inside a medical box.
Clinical samples include thoseClinical samples include those from humans and animals.
57
5.Other regulations
58
Recombinant gene experiments(Act on the Conservation and Sustainable Use of Biological Diversity through R l ti th U f Li i M difi d O i )Regulations on the Use of Living Modified Organisms)
1.Class 2, 3 and 4 organism include pathogensTh l ifi ti diff t f th f K h h
Living modified organism including pathogen genes have to be used in P2 or P3 according to the levels
These classifications are different from those of Kansenshouhou
2.Laboratory containment levels are called P1 to 3, but
used in P2 or P3 according to the levels.
not BSL-1 to 3Required registrations of each laboratory for recombinant gene experiments
3.Minister confirmation experiments
experiments
① in case the host of Living modified organism is “virus”② in case the Living modified organism includes class 4 organism③ in case an unlisted organism is used for the recombinant gene
experiments.59
Summary
・Complete required registrations before experimentsComplete required registrations before experiments
・Practice proper biosafety and biosecurity
・Decide rules for each laboratory and train their staffs
・Please contact with Kenkyu-kikakuka if you havequestions
60