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Biosafety/Bloodborne Pathogens:
Working Safely With
Biological Materials in Laboratories
http://research.wayne.edu/oehs
What is a Biohazard?
A biohazard is an agent of biological origin that
has the capacity to produce deleterious effects
on ...
Laboratories working with biohazards have a set of guidelines to
follow from the Centers for Disease Control (CDC) and the...
Research using biohazards requires
review and approval by the
WSU Institutional Biosafety Committee (IBC)
 An IBC is mand...
The principal investigator must submit the
appropriate form to the committee:
 Exempt rDNA research does not need approva...
The appropriate biosafety level (1-4) is
determined by the CDC/NIH.
 Selection of the biosafety level is based on
charact...
Biosafety Levels
 BSL1 - agents not known to cause disease.
 BSL2 - agents associated with human disease.
 BSL3 - indig...
Biosafety Levels 1- 4 provide:
 Increasing levels of personnel & environmental protection,
and appropriate guidelines for...
This presentation will focus on
the CDC/NIH guidelines for
biosafety levels 1 – 2 research
laboratories.
8
Proper handwashing is
important at all
biosafety levels!
 Wash with warm, running water, mild, preferably liquid
soap, (d...
 LAB COAT
 protects clothing/skin
 GLOVES:
 disposable latex/non-latex exam gloves
 change when torn or contaminated
...
 If risk of splashes or aerosols,
protect the eyes & face.
 safety glasses/goggles
 face mask
 If necessary, other PPE...
Biosafety Level 1 (BSL1)
 BSL 1 is suitable for work involving well-characterized
agents not known to cause disease in he...
Biosafety Level 2 (BSL 2)
 BSL 2 is suitable for work involving agents of moderate
potential hazard to personnel and the ...
All Biosafety Level 2 (or higher) labs
are inspected to assure they meet the
CDC/NIH guidelines.
14
Biosafety Levels 1 – 2
Facility Design Guidelines
 Lab doors are lockable.
 Sinks available for hand washing.
 Work sur...
Biosafety Level 2 Additional Requirements
 Biosafety Cabinet
 Autoclave
 Eye Wash 16
Biosafety Levels 1 – 2
Standard Microbiological Practices
 Restrict/limit access when working
 No eating, drinking, stor...
 Extreme precaution with
SHARPS
 Gloves and additional PPE
 Use of mechanical pipetting
devices
18
Biosafety Levels 1 –...
 Extra care should be taken when using any contaminated
sharp item, including needles and syringes, slides, pipettes,
cap...
 Used disposable needles must not be bent, sheared,
broken, recapped, removed from disposable syringes, or
otherwise mani...
 Inspection by the Biosafety Officer
 WSU Biosafety Manual available.
 Biosafety SOPs are read & signed by all staff.
...
Hazard warning signs
Include the following:
• Entry requirements
• Agent name/Human cells
• Equipment
• BSC
• Refrigerator...
Hazard warning signs
 Risk Group 2 Agents
23
 Universal Precautions
 Labs using human/non-human
primate cells, cell lin...
 Cultures, tissue, specimens should be placed in a container
with a cover that prevents leakage during collection,
handli...
MIOSHA Occupational Exposure to
Bloodborne Diseases Standard
R325.0001-325.0018
 Standard applies to all jobs where there...
Use of human materials in research
 BSL-2 is appropriate for activities with all primate cell lines,
even well establishe...
Use of human materials in research
 Laboratories using human cell strains (non-transformed
cells) propagated from primary...
Bloodborne Diseases
 HIV: Human Immunodeficiency Virus
causes AIDS - no cure or vaccination
28
 HBV: Hepatitis B virus c...
Bloodborne diseases are only spread when blood and
certain other body fluids from an infected source get into
the bloodstr...
Bloodborne Diseases are NOT
transmitted through:
 Kissing or hugging
 Sneezing or coughing
 Food or water
 Sharing eat...
How is HIV Transmitted?
 Blood and body fluids
 serum
 semen
 vaginal secretions
 fluids around internal organs/syste...
HIV Transmission in Healthcare
As of December 31, 2013, only 58
confirmed occupational transmissions and
150 possible tran...
Healthcare workers with documented & possible occupationally acquired
AIDS/HIV infection, by occupation, 1985 - 2013.
Occu...
HIV Exposure Risk
 Rate of seroconversion after needlestick exposure
to infective material from HIV+ person is 0.3% or
ab...
HIV Transmission in
Healthcare Workers
 Factors associated with HIV transmission in healthcare:
 Deep injury
 Device vi...
Diagnoses of HIV Infection among Adults and
Adolescents, by Transmission Category, 2014—United
States and 6 Dependent Area...
Hepatitis B Virus
 Inflammation of the liver – most
common bloodborne disease
 Symptoms range from flu-like to
none at a...
Hepatitis B Facts
 Incubation period from 28-160 days
 Symptoms may include:
 loss of appetite
 fatigue
 fever
 poss...
 Fluids that pose risk of infection:
 blood and blood products
 body fluids containing visible blood
 semen and vagina...
How can Hepatitis B be transmitted
in the workplace?
 Needlsticks or cuts with sharp, contaminated
objects
 splashes to ...
Hepatitis B in Healthcare Workers
(HCWs) in the U.S.
 Prior to vaccination, it was estimated that more than
12,000 HCWs w...
Source: National Notifiable Diseases Surveillance System (NNDSS)
 Administered in 3 shots over 6 months.
 Engerix-B Vaccine is yeast derived - no chance of infection
from vaccination
 ...
Hepatitis B Vaccination
 Vaccination is given at 4K University Health Center (Detroit
Receiving Hospital), Monday – Frida...
Hepatitis C Virus
 Spread primarily through
blood/blood products. Less
likely to be spread sexually.
 If left untreated,...
Hepatitis C Virus
 Identified in 1988, formerly called non-A non-B
hepatitis - called “silent epidemic”
 Blood supply no...
Incidence of HCV infection in U.S.
 Est. number of chronic HCV cases in the U.S. is 2.7 – 3.9 million.
 Approximately 75...
Source: CDC, National Notifiable Diseases Surveillance System (NNDSS)
Source: CDC, National Notifiable Diseases Surveillance System (NNDSS)
*A total of 2,194 case reports of acute hepatitis C ...
Generation of Aerosols
 If aerosols may be generated, work must be performed
in Class II biosafety cabinet.
 Procedures ...
Biological Safety Cabinets
these cabinets provide:
 Product protection
 Personal protection
 Environmental protection
51
Laminar Flow “Clean Benches”
 This is not a Biosafety Cabinet –
it does NOT protect the
worker!
 Filtered air flows from...
Biological Safety Cabinets
 Contain HEPA Filters (high efficiency
particulate air). These trap particulates
as small as 0...
Biological Safety Cabinets
 May use for non-volatile toxic
chemicals or low-level radioactive
materials.
 May use for “m...
Biological Safety Cabinets
Working Safely
 Disinfect work surfaces with iodine based
disinfectant (e.g. Wescodyne).
 Loa...
Biological Safety Cabinets
Working Safely
 Always enter straight into
cabinet – no sweeping
motions.
 Perform work in a ...
 When finished, decontaminate all
items to be removed from
cabinet.
 Decon work surfaces again.
 Allow cabinet to run 1...
Natural Gas in a BSC
 Natural gas/Bunsen Burner use in a BSC represents a significant
fire risk.
 Use of flames & excess...
Safe Centrifuge Use
 Check tubes for cracks/chips.
 Use matched sets of tubes,
buckets, etc.
 Tightly seal all tubes an...
Proper vacuum system set-up
A. Primary flask – type of disinfectant
B. Secondary flask – overfill – should be cleaned imme...
Emergency Response
Surface Decontamination
 Alert co-workers
 Define/isolate contaminated area
 Put on appropriate PPE
...
Emergency Response
Surface Decontamination
 If using bleach, mix FRESH 1:9 solution.
 Allow adequate contact time (20 mi...
 Minimum strength of cleaners must be tuberculocidal
(kills M. tuberculosis var. bovis and all vegetative
bacteria, fungi...
Emergency Response
responding to exposures
 Occupational exposure is contact with blood or other
potentially infectious m...
 Thoroughly wash affected area. Do not wash with bleach or
other strong cleaners.
 If eyes/face exposed, flush in eyewas...
66
Biological Waste Disposal
Request Waste Pick-Ups & Replacement
Containers on-line at:
http://research.wayne.edu/oehs/bio-s...
 Must be used for all SHARPS
(contaminated or not)
 Don’t overfill containers!
 Locate containers conveniently.
 Never...
 Hypodermic needles, with syringe
 IV tubing with needles attached
 Contaminated Pasteur pipettes
 Razors and microtom...
NEVER dispose of SHARPS
in glass waste boxes or in the trash!
70
Biological Waste (Red) Bins
 Free 28 gallon red plastic bins and a
liner bag are provided by OEH&S.
 Labs must purchase ...
 Items contaminated with human or animal blood, body
fluids or tissue.
 Cultures/stocks of infectious agents: including ...
 When bin is full:
 close & tie liner bag shut
 place lid on securely
 do not overfill bins. They shouldn’t weigh more...
All biohazard waste must be in
secondary containment
 Biohazard bags must always
be stored in leakproof
secondary contain...
Improperly stored waste
Properly stored waste
• Closeable
• Constructed to contain all
contents and prevent leakage
of fluids
• Puncture resistant...
Waste glass & plastic disposal
 Decontaminate items with
bleach or autoclave before
putting into box.
 Use a sturdy, dur...
78
 When box is full, tape shut
securely.
 Place box in an area for the
custodial staff to dispose.
 Custodians reserve...
Autoclave Decontamination
 Place items in secondary containers:
stainless steel or autoclavable
plastic bins.
 Temps of ...
 Do not cap or plug vessels.
 Do not add excessive amounts of liquid
to load.
 Usually steam remains in the chamber
at ...
Liquid Biological Waste Disposal
 Aqueous biological solutions such as blood, urine, cells, microbial
cultures must be au...
Working with research animals
 Animals release airborne allergens which can cause
allergy symptoms in some people.
 Mild...
Shipping Dangerous Goods
• Common materials shipped include: human
and animal samples, bacteria, DNA, proteins,
chemicals ...
Take the Quiz!
To verify your completion of this course through WSU,
you must return to the CITI site and take the quiz.
8...
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Biosafety/Bloodborne Pathogens Training for Laboratory Workers

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The Biosafety/Bloodborne Pathogens Course is required for all Wayne State University investigators, staff, and students who work in a lab with materials that are potentially infectious, including human blood, body fluids, tissue, cell lines, animals infected with human pathogens, mammalian viruses, or any agents that are handled at Biosafety Level 2 (BSL2).

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Biosafety/Bloodborne Pathogens Training for Laboratory Workers

  1. 1. Biosafety/Bloodborne Pathogens: Working Safely With Biological Materials in Laboratories http://research.wayne.edu/oehs
  2. 2. What is a Biohazard? A biohazard is an agent of biological origin that has the capacity to produce deleterious effects on humans, e.g.; microorganisms, toxins and allergens derived from those organisms; and allergens and toxins derived from higher plants and animals. 1
  3. 3. Laboratories working with biohazards have a set of guidelines to follow from the Centers for Disease Control (CDC) and the National Institutes of Health (NIH):  The guidelines are in place to protect human health and prevent the release of pathogens into the environment.  The guidelines include:  laboratory practices and procedures  facility design  safety equipment requirements Link to the CDC/NIH BMBL 5th Edition 2
  4. 4. Research using biohazards requires review and approval by the WSU Institutional Biosafety Committee (IBC)  An IBC is mandated by the NIH at every institution.  IBC members include WSU faculty, staff and PhDs from neighboring institutions.  The IBC may increase the Biosafety Level requirements if the vector is more infectious than wild-type virus. 3
  5. 5. The principal investigator must submit the appropriate form to the committee:  Exempt rDNA research does not need approval - researchers must sign off on exemption form. Protocols that require IBC review & approval include:  Native or recombinant mammalian viruses  Pathogenic/infectious agents (non-rDNA)  Biological toxins or rDNA encoding a toxin  Work with, or creation of, transgenic animals. WSU Biological Agent User Application Forms 4
  6. 6. The appropriate biosafety level (1-4) is determined by the CDC/NIH.  Selection of the biosafety level is based on characteristics of the infectious agent:  Pathogenicity of material - disease incidence/severity  Documented route of transmission (bloodborne, airborne, ingestion)  Availability of protective immunization (HBV Vaccine) or effective therapy  Risk of exposure created by manipulation in handling the agent & caring for infected animals 5
  7. 7. Biosafety Levels  BSL1 - agents not known to cause disease.  BSL2 - agents associated with human disease.  BSL3 - indigenous/exotic agents associated with human disease and with potential for aerosol transmission.  BSL4 - dangerous/exotic agents of life threatening nature. 6
  8. 8. Biosafety Levels 1- 4 provide:  Increasing levels of personnel & environmental protection, and appropriate guidelines for:  Laboratory Practices and Techniques  Standard Practices and Special Practices  Knowledge of supervisor and personnel  Lab specific SOPs/Biosafety manual  Safety Equipment (Primary Barriers)  Laboratory Facilities (Secondary Barriers)  Buildings (Tertiary Barriers) 7
  9. 9. This presentation will focus on the CDC/NIH guidelines for biosafety levels 1 – 2 research laboratories. 8
  10. 10. Proper handwashing is important at all biosafety levels!  Wash with warm, running water, mild, preferably liquid soap, (doesn’t have to be antibacterial).  Rub hands together vigorously for at least 15 seconds: scrub between fingers, under nails, tops & palms of hands.  Rinse with warm, running water.  Dry with disposable paper towel.  Use lotion to prevent chapping of hands. 9
  11. 11.  LAB COAT  protects clothing/skin  GLOVES:  disposable latex/non-latex exam gloves  change when torn or contaminated Basic Personal Protective Equipment (PPE) is required at all Biosafety Levels 10 Remove PPE before leaving the work area!
  12. 12.  If risk of splashes or aerosols, protect the eyes & face.  safety glasses/goggles  face mask  If necessary, other PPE should be worn, including:  gown, face shield, booties, etc. Basic Personal Protective Equipment (PPE) is required at all Biosafety Levels 11
  13. 13. Biosafety Level 1 (BSL1)  BSL 1 is suitable for work involving well-characterized agents not known to cause disease in healthy adults and of minimal potential hazard to lab personnel and the environment. Some examples include:  Bacillus subtilis  Naegleria gruberi  Infectious canine hepatitis virus  Non-entero hemorrhagic E. coli  Exempt recombinant DNA experiments 12
  14. 14. Biosafety Level 2 (BSL 2)  BSL 2 is suitable for work involving agents of moderate potential hazard to personnel and the environment.  Immunization or antibiotic treatment is available  Examples include:  Measles virus  Salmonellae  Toxoplasma spp.  Hepatitis B virus  Adenoviruses 13
  15. 15. All Biosafety Level 2 (or higher) labs are inspected to assure they meet the CDC/NIH guidelines. 14
  16. 16. Biosafety Levels 1 – 2 Facility Design Guidelines  Lab doors are lockable.  Sinks available for hand washing.  Work surfaces easily cleaned, impervious to water. 15  BSL2 labs should be under negative pressure. Air flows into lab and doesn’t re-circulate to non-lab areas.
  17. 17. Biosafety Level 2 Additional Requirements  Biosafety Cabinet  Autoclave  Eye Wash 16
  18. 18. Biosafety Levels 1 – 2 Standard Microbiological Practices  Restrict/limit access when working  No eating, drinking, storing food, etc.  No mouth pipetting  Minimize splashes and aerosols  Decontaminate wastes  Decontaminate work surfaces daily  Maintain insect & rodent control program 17
  19. 19.  Extreme precaution with SHARPS  Gloves and additional PPE  Use of mechanical pipetting devices 18 Biosafety Levels 1 – 2 Standard Microbiological Practices +
  20. 20.  Extra care should be taken when using any contaminated sharp item, including needles and syringes, slides, pipettes, capillary tubes, razor blades, lancets, and scalpels.  Plastic should be substituted for glass whenever possible. 19 Biosafety Level 2 Precautions with SHARPS
  21. 21.  Used disposable needles must not be bent, sheared, broken, recapped, removed from disposable syringes, or otherwise manipulated by hand before disposal.  Always dispose of whole unit in SHARPS containers! 20 Biosafety Level 2 Precautions with SHARPS
  22. 22.  Inspection by the Biosafety Officer  WSU Biosafety Manual available.  Biosafety SOPs are read & signed by all staff.  Labs using materials from human source must review and sign off on WSU Bloodborne Diseases Exposure Control Plan.  Principal Investigator must train staff annually. 21 Biosafety Level 2 Additional Requirements
  23. 23. Hazard warning signs Include the following: • Entry requirements • Agent name/Human cells • Equipment • BSC • Refrigerators/freezers • Incubators • Regulated waste
  24. 24. Hazard warning signs  Risk Group 2 Agents 23  Universal Precautions  Labs using human/non-human primate cells, cell lines, organ cultures and body fluids – doesn’t require IBC approval.  Require IBC Approval
  25. 25.  Cultures, tissue, specimens should be placed in a container with a cover that prevents leakage during collection, handling, processing, transport, storage or shipping. 24 Biosafety Level 2 Additional precautions
  26. 26. MIOSHA Occupational Exposure to Bloodborne Diseases Standard R325.0001-325.0018  Standard applies to all jobs where there is a potential for exposure to human bloodborne diseases, including healthcare workers, laboratory workers, firefighters, EMTs, custodians and maintenance workers, etc. 25  Retraining is required annually!  Based on the concept of UNIVERSAL PRECAUTIONS: treating all human blood, tissue, cells, etc. as if they ARE infectious.
  27. 27. Use of human materials in research  BSL-2 is appropriate for activities with all primate cell lines, even well established ones, all cells derived from primate lymphoid or tumor tissues; all primate tissue; all human clinical material*; cultured cells new to the laboratory until proven contaminant-free; and, cells exposed to or transformed by a primate oncogenic virus. * These activities and the use of any cells purposely infected with or suspected of harboring agents defined as bloodborne pathogens are covered by the Bloodborne Pathogens Standard) and WSU’s Exposure Control Plan.
  28. 28. Use of human materials in research  Laboratories using human cell strains (non-transformed cells) propagated from primary explants must also comply with the Bloodborne Pathogens Standard) and WSU’s Exposure Control Plan because they are considered “unfixed human tissue” and may carry bloodborne diseases such as Hepatitis B, Hepatitis C, HIV.
  29. 29. Bloodborne Diseases  HIV: Human Immunodeficiency Virus causes AIDS - no cure or vaccination 28  HBV: Hepatitis B virus causes liver disease vaccination available  HCV: Hepatitis C virus causes liver disease no vaccination available
  30. 30. Bloodborne diseases are only spread when blood and certain other body fluids from an infected source get into the bloodstream of an uninfected person. An occupational exposure is contact with infected material through:  needlestick or cut with contaminated sharp object  cuts/breaks in skin  mucous membranes (eyes, nose, mouth) 29
  31. 31. Bloodborne Diseases are NOT transmitted through:  Kissing or hugging  Sneezing or coughing  Food or water  Sharing eating utensils, cups, etc.  Casual contact  Saliva, tears, perspiration, urine, feces (when there is no blood present)
  32. 32. How is HIV Transmitted?  Blood and body fluids  serum  semen  vaginal secretions  fluids around internal organs/systems  IV drug use  vaginal or anal intercourse  mother to child in utero * There are no documented cases of HIV being transmitted through saliva, tears or perspiration, saliva. 31
  33. 33. HIV Transmission in Healthcare As of December 31, 2013, only 58 confirmed occupational transmissions and 150 possible transmissions of HIV to healthcare workers had been reported in the United States. Reporting of this data began in 1985. (Only one confirmed case has been reported since 1999) http://www.cdc.gov/hiv/resources/factsheets/PDF/hcw.pdf
  34. 34. Healthcare workers with documented & possible occupationally acquired AIDS/HIV infection, by occupation, 1985 - 2013. Occupation Documented Possible Nurse 24 37 Laboratory worker, clinical 16 21 Physician, nonsurgical 6 13 Laboratory technician, nonclinical 4 - Housekeeper/maintenance worker 2 14 Technician, surgical 2 2 Embalmer/morgue technician 1 2 Health aide/attendant 1 16 Respiratory therapist 1 2 Technician, dialysis 1 3 Dental worker, including dentist - 6 Emergency medical technician/paramedic - 13 Physician, surgical - 6 Other technician/therapist - 9 Other healthcare occupation - 6 Total 58 150
  35. 35. HIV Exposure Risk  Rate of seroconversion after needlestick exposure to infective material from HIV+ person is 0.3% or about 1 in 300.  HIV in high concentration during period prior to antibody development.  Much less infective than HBV, HCV, Herpes Source: Centers for Disease Control 34
  36. 36. HIV Transmission in Healthcare Workers  Factors associated with HIV transmission in healthcare:  Deep injury  Device visibly contaminated with source patient’s blood  Procedures involving a needle placed directly in a vein or artery  Terminal illness in source patient  No zidovudine (AZT) prophylaxis 35
  37. 37. Diagnoses of HIV Infection among Adults and Adolescents, by Transmission Category, 2014—United States and 6 Dependent Areas N = 44,609 Note. Data include persons with a diagnosis of HIV infection regardless of stage of disease at diagnosis. All displayed data have been statistically adjusted to account for reporting delays and missing transmission category, but not for incomplete reporting. a Heterosexual contact with a person known to have, or to be at high risk for, HIV infection. b Includes hemophilia, blood transfusion, perinatal exposure, and risk factor not reported or not identified.
  38. 38. Hepatitis B Virus  Inflammation of the liver – most common bloodborne disease  Symptoms range from flu-like to none at all  No symptoms – person can still be infectious and can spread the disease 37
  39. 39. Hepatitis B Facts  Incubation period from 28-160 days  Symptoms may include:  loss of appetite  fatigue  fever  possible jaundice and dark urine  HBV is a much greater risk on the job than HIV, especially if you are not vaccinated. 38
  40. 40.  Fluids that pose risk of infection:  blood and blood products  body fluids containing visible blood  semen and vaginal secretions  breast milk  saliva (through a human bite) Hepatitis B Facts 39
  41. 41. How can Hepatitis B be transmitted in the workplace?  Needlsticks or cuts with sharp, contaminated objects  splashes to eyes/nose/mouth  contact with broken skin  human bites that break the skin 40
  42. 42. Hepatitis B in Healthcare Workers (HCWs) in the U.S.  Prior to vaccination, it was estimated that more than 12,000 HCWs were occupationally infected with HBV annually, resulting in 250 deaths.  1983 – Incidence of HBV among HCWs was 3 times higher than in the general population.  By 1995 it was 5 times lower.  Advent of HBV vaccine was a major advance in preserving health and lives of HCWs. Source: Arch Intern Med 1997; 157:2601-2605 41
  43. 43. Source: National Notifiable Diseases Surveillance System (NNDSS)
  44. 44.  Administered in 3 shots over 6 months.  Engerix-B Vaccine is yeast derived - no chance of infection from vaccination  Provides long term protection against HBV for 96% of healthy adults: no booster recommendation by CDC at this time.  Post exposure vaccination is 70-88% effective when started within one week. 43 Hepatitis B Vaccination
  45. 45. Hepatitis B Vaccination  Vaccination is given at 4K University Health Center (Detroit Receiving Hospital), Monday – Friday 9:00 – 4:30.  Must present certificate of training to be vaccinated.  Must sign declination form if you choose to not be vaccinated.  Call 313-577-1200 for more information. 44
  46. 46. Hepatitis C Virus  Spread primarily through blood/blood products. Less likely to be spread sexually.  If left untreated, 80-85% of cases become chronic  There is no vaccine for HCV. Based on limited studies, risk for infection after needlestick is approximately 1.8%. 45
  47. 47. Hepatitis C Virus  Identified in 1988, formerly called non-A non-B hepatitis - called “silent epidemic”  Blood supply not tested until early 90s.  Most common chronic bloodborne infection in U.S.  Incubation period from 2-26 weeks. Most people never have symptoms 46
  48. 48. Incidence of HCV infection in U.S.  Est. number of chronic HCV cases in the U.S. is 2.7 – 3.9 million.  Approximately 75% are chronically infected and may not be aware of their infection because they are not clinically ill.  Persons newly infected with HCV are usually asymptomatic, so acute hepatitis C is rarely identified or reported.  Most prevalent among those born during 1945–1965, who were likely infected during the 1970s and 80s when rates were highest. 47 Reported Acute (New) Cases of Hepatitis C Virus (HCV) Estimated Actual New Cases of HCV (range) in 2011* 2005 2006 2007 2008 2009 2010 2011 2011 (estimated)* 694 802 849 878 781 853 1,229 16,500 (7,200- 43,400) * Actual acute cases estimated to be 13.4 times the number of reported cases in any year
  49. 49. Source: CDC, National Notifiable Diseases Surveillance System (NNDSS)
  50. 50. Source: CDC, National Notifiable Diseases Surveillance System (NNDSS) *A total of 2,194 case reports of acute hepatitis C were received in 2014. † More than one risk exposure/behavior may be indicated on each case-report. §Risk data not reported. ¶A total of 1,174 acute hepatitis C cases were reported among males in 2014.
  51. 51. Generation of Aerosols  If aerosols may be generated, work must be performed in Class II biosafety cabinet.  Procedures that may generate aerosols include:  Pipetting  Centrifugation  Tissue culture  Sonication  Blending of tissues  Animal innoculations 50
  52. 52. Biological Safety Cabinets these cabinets provide:  Product protection  Personal protection  Environmental protection 51
  53. 53. Laminar Flow “Clean Benches”  This is not a Biosafety Cabinet – it does NOT protect the worker!  Filtered air flows from back of cabinet, across work surface, and towards user.  Protects samples and work but NOT the user 52
  54. 54. Biological Safety Cabinets  Contain HEPA Filters (high efficiency particulate air). These trap particulates as small as 0.3u.  Does not protect from chemicals: fumes and vapors pass through  Chemicals and heat may damage filter. 53
  55. 55. Biological Safety Cabinets  May use for non-volatile toxic chemicals or low-level radioactive materials.  May use for “minute” amounts of volatile chemicals.  Ensure annual certification from OEH&S.  Place BSC away from high traffic areas, airflow ducts and lab entrance doors. 54
  56. 56. Biological Safety Cabinets Working Safely  Disinfect work surfaces with iodine based disinfectant (e.g. Wescodyne).  Load cabinet with all needed supplies.  Allow cabinet to run 10-15 minutes before beginning work.  Check inward airflow with a piece of tissue. 55
  57. 57. Biological Safety Cabinets Working Safely  Always enter straight into cabinet – no sweeping motions.  Perform work in a slow, methodical manner.  Place materials well within the cabinet – don’t block grills.  Place discard pan within cabinet – discard pipettes inside in disinfectant tray 56
  58. 58.  When finished, decontaminate all items to be removed from cabinet.  Decon work surfaces again.  Allow cabinet to run 10-15 minutes before turning off.  Only use UV light as secondary disinfection. 57 Biological Safety Cabinets Working Safely
  59. 59. Natural Gas in a BSC  Natural gas/Bunsen Burner use in a BSC represents a significant fire risk.  Use of flames & excessive heat can compromise the integrity of the BSC and increase exposure risks.  Explore safer options, including:  Disposable loops/spreaders  Electric Bunsen Burners  Electric incinerators  Hot bead sterilizers 58
  60. 60. Safe Centrifuge Use  Check tubes for cracks/chips.  Use matched sets of tubes, buckets, etc.  Tightly seal all tubes and safety cups.  Ensure that rotor is locked to spindle and bucket seated.  Close lid during operation.  Allow to come to complete stop before opening. 59
  61. 61. Proper vacuum system set-up A. Primary flask – type of disinfectant B. Secondary flask – overfill – should be cleaned immediately C. Hepa filter – component that is frequently missing! HEPA filters: Whatman HEPA-Vent Filter Inlet/Outlet: Fisher Scientific Catalog #: 09-744-79
  62. 62. Emergency Response Surface Decontamination  Alert co-workers  Define/isolate contaminated area  Put on appropriate PPE  Remove glass with forceps or scoop  Apply absorbent towel(s) to spill; remove bulk & reapply if needed  Apply disinfectant on top of towels. 61
  63. 63. Emergency Response Surface Decontamination  If using bleach, mix FRESH 1:9 solution.  Allow adequate contact time (20 minutes).  Remove towel & mop up.  Clean again with soap/water or alcohol.  Properly dispose of materials in biohazard bins.  Wash your hands last.  For larger spills or questions, contact OEHS. 62
  64. 64.  Minimum strength of cleaners must be tuberculocidal (kills M. tuberculosis var. bovis and all vegetative bacteria, fungi, and most viruses)  Require minimum 20 minute exposure  Examples: quaternary ammonium detergents, phenolics, iodophores, chlorine compounds, alcohols 63 Emergency Response Surface Decontamination – other acceptable disinfectants
  65. 65. Emergency Response responding to exposures  Occupational exposure is contact with blood or other potentially infectious materials:  needlestick or cut with sharp, contaminated object  contact with broken skin through cuts, rashes, other breaks in skin  splashes to eyes, nose, mouth 64
  66. 66.  Thoroughly wash affected area. Do not wash with bleach or other strong cleaners.  If eyes/face exposed, flush in eyewash immediately.  Report incident to supervisor: Get medical attention! 4K University Health Center: includes assessment, blood tests, titer, counseling, follow-up visits…all confidential.  Complete a WSU Report of Injury form: Risk Management 313-577-3110. 65 Emergency Response responding to exposures
  67. 67. 66
  68. 68. Biological Waste Disposal Request Waste Pick-Ups & Replacement Containers on-line at: http://research.wayne.edu/oehs/bio-safety/biological-waste.php SHARPS containers Red Bins 67
  69. 69.  Must be used for all SHARPS (contaminated or not)  Don’t overfill containers!  Locate containers conveniently.  Never recap needles: #1 cause of needlesticks! Correct SHARPS Disposal Get free containers from OEH&S  Request SHARPS pick-up and replacement containers on OEH&S website:  http://research.wayne.edu/oehs/bio-safety/biological-waste.php 68
  70. 70.  Hypodermic needles, with syringe  IV tubing with needles attached  Contaminated Pasteur pipettes  Razors and microtome blades  Scalpels  Lancets 69 Correct SHARPS Disposal What goes into SHARPS container?
  71. 71. NEVER dispose of SHARPS in glass waste boxes or in the trash! 70
  72. 72. Biological Waste (Red) Bins  Free 28 gallon red plastic bins and a liner bag are provided by OEH&S.  Labs must purchase their own autoclavable biohazard bags  Some materials should be autoclaved before placing into bins. Check with the Biosafety Officer for more information.  Always place bags in bins or other leak-proof secondary containers.  DO NOT autoclave bin or the liner bag! 71
  73. 73.  Items contaminated with human or animal blood, body fluids or tissue.  Cultures/stocks of infectious agents: including waste from production of biologicals, discarded vaccines, and culture dishes.  Materials/microorganisms used in recombinant DNA research.  NO SHARPS (needles, razor blades, etc!) 72 Biological Waste (Red) Bins what goes into bins?
  74. 74.  When bin is full:  close & tie liner bag shut  place lid on securely  do not overfill bins. They shouldn’t weigh more than 35 lbs.  request pick-up and replacement containers from OEH&S website: http://research.wayne.edu/oehs/bio-safety/biological-waste.php Only properly prepared bins will be accepted! 73
  75. 75. All biohazard waste must be in secondary containment  Biohazard bags must always be stored in leakproof secondary containers 74
  76. 76. Improperly stored waste
  77. 77. Properly stored waste • Closeable • Constructed to contain all contents and prevent leakage of fluids • Puncture resistant • Labeled with biohazard symbol
  78. 78. Waste glass & plastic disposal  Decontaminate items with bleach or autoclave before putting into box.  Use a sturdy, durable box.  Label the box “broken glass” or “glass waste”  Line the box with plastic bag.  Keep weight reasonable – 25 lbs. or less! 77
  79. 79. 78  When box is full, tape shut securely.  Place box in an area for the custodial staff to dispose.  Custodians reserve the right to refuse pick up of boxes that aren’t prepared properly. Waste glass & plastic disposal
  80. 80. Autoclave Decontamination  Place items in secondary containers: stainless steel or autoclavable plastic bins.  Temps of 121 degrees C for 20 minutes for most recombinant organisms and pathogens.  Larger loads require more time.  Autoclaves are usually in shared areas – clean up after yourself! 79
  81. 81.  Do not cap or plug vessels.  Do not add excessive amounts of liquid to load.  Usually steam remains in the chamber at end of a liquid cycle, so be careful!  Wear eye and face protection.  Stand behind door when opening it – keep face away from door.  Slowly open door only a crack to allow residual steam to escape. 80 Autoclave Decontamination
  82. 82. Liquid Biological Waste Disposal  Aqueous biological solutions such as blood, urine, cells, microbial cultures must be autoclaved or chemically disinfected (treated with bleach)  1 part bleach to 9 parts contaminated liquid – let stand for 30 minutes.  After treating, dispose down drain with lots of H2O.  Do not autoclave bleach treated waste. 81
  83. 83. Working with research animals  Animals release airborne allergens which can cause allergy symptoms in some people.  Mild symptoms include: itchy/watery eyes, runny nose, skin rash/itchy skin, nasal congestion, and cough.  Reduce your exposure by using PPE, including: Gloves Lab Coat N-95 Mask for your nose/mouth (requires a fit-test)  For more information on WSU’s Animal Contact program, call the OEH&S Occupational Health Nurse at 577-5917. 82
  84. 84. Shipping Dangerous Goods • Common materials shipped include: human and animal samples, bacteria, DNA, proteins, chemicals (solids or liquids), dry ice, etc. • Shipping ANY material on dry ice by air is considered a Dangerous Good/Hazardous Material • OEH&S is the designated authority for shipping Dangerous Goods from WSU. • Contact OEH&S at 577-1200 to ship any Dangerous Goods.
  85. 85. Take the Quiz! To verify your completion of this course through WSU, you must return to the CITI site and take the quiz. 84 If you are unable to take the quiz on CITI, email fracassa@wayne.edu.

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