USC Environmental Health & Safety

GoalSafety: 385,000 percutaneous injuries/yr in hospitals1990 OSHA estimate: 9,000 bloodborne infections/yr, 200

deathsLaboratory acquired infections - Sulkin and Pike: 3,921

cumulative cases, including brucellosis, typhoid, tularemia, tuberculosis, hepatitis, and venezuelan equine encephalitisLess than 20% associated with known accidentLab environment more hazardous than nature due to

culturing, propagation of infectious agentsRecent events at U Chicago, Texas A&MCompliance with Safety Regulations: Cal-OSHA BBP

Standard, 1992Education, prevention

Lab acquired vaccinia infection, Virginia 2008

Worked in cancer research labHandled mice infected with vaccinia virusNot vaccinated

What are Bloodborne Pathogens?microorganisms ( such as viruses or bacteria)

that are carried in blood and can cause disease in people

malaria, syphilis, Hepatitis B Virus(HBV), Hepatitis C Virus, Human Immunodeficiency Virus (HIV)

Am I at risk?Work with blood or OPIMOPIM - semen, vaginal secretions, saliva in

dental procedures, CSF, or other internal body fluidsOrgans, unfixed human tissueHBV, HCV, HIV tissue culturesHuman cell lines

Human Cell LinesCell lines may be infected or become

infected/contaminated in subsequent handling/passaging

LCMV- Researchers infected working with nude mice (infected tumor cell line)

“We recommend that all human cell lines be accorded the same level of biosafety consideration as a line known to carry HIV… Thus, it is best to use caution when handling any human cell line”

Universal Precaution

Low Risk MaterialsUrine, feces, vomit, tears, sweat, sputum,

nasal secretionsUnless visibly contaminated with blood

Routes Of TransmissionSexual contactSharing needlesBlood Transfusion (very rarely in countries where

blood is screened)Birth: Expose through amniotic fluid, blood during the

birthing process, and (less likely) through breast milk Accidental puncture from contaminated

needles, glass or other sharpsContact between broken or damaged skin and

infected body fluidsContact between mucous membranes and infected

body fluids

Common procedures where exposure may occurHandling containers of blood, fluid, tissue,

or culturesPhlebotomyPipetting, mixing, or handling blood, fluid, or

tissueCleaning blood/body fluid spillsHandling contaminated sharps or other

contaminated wastePuncture from improperly disposed needle

Injections/inoculations~89% occupationally acquired HIV

Determinants of Disease TransmissionIndividual receiving the doseVirulence / type of pathogenSize of delivered dose (concentration)Route of exposureType of bodily fluid Whether post exposure prophylaxis was

administered

Diseases caused by BBPHepatitis BHepatitis CAIDS(HIV)

Hepatitis BInflammation of the liverRisk of HBV infection after a single positive needle stick is 30%Durable virus (7 days outside body)Acute or chronicMore than 350 million people are chronic carriers of HBV

worldwide (CDC) 800,000–1.4 million persons in the United States

have chronic HBV infection

Annual number of unreported infections may be 10 times greater than the number of reported (many HBV infections are either asymptomatic or never reported)

rate of new HBV infections has declined ~80% since 1991

Hepatitis BSymptoms (weeks-~6months) – Fever,

Jaundice, fatigue, abdominal pain, loss of appetite, nausea, vomiting, joint pain, dark urine

Complications - Cirrhosis (scarring) of the liver, liver cancer, liver failure, and death

Prevention - Hepatitis B Vaccine, PEP (Plasma fractionation of Hep B Antibody ), Universal Precautions

http://phil.cdc.gov/phil/home.asp

HEP B VaccineUSC offers the vaccines to employees

FREE OF CHARGESeries of 3 inoculations (shots) and Titer

checkAfter vaccination cellular immunity

persists (despite low antibody levels)

Hepatitis CAfter a needlestick or sharps exposure to

HCV-positive blood, the risk of HCV infection is approximately 1.8%

75-80% chronic 3.2 million persons in the U.S.- chronic HCV

infectionVaccine? NoPEP? No

Hepatitis C VirusOf every 100 persons infected with HCV,

approximately 75–85 will go on to develop chronic infection60–70 will go on to develop chronic liver disease5–20 will go on to develop cirrhosis over a period of 20–30 years 1–5 will die from the consequences of chronic infection (liver cancer or cirrhosis)

8,000–10,000 deaths each year in the United States (chronic)

Hepatitis C Symptoms - Jaundice, fatigue, abdominal

pain, loss of appetite, nausea, dark urineComplications – Chronic liver disease, deathTransmission: Occurs when blood or body

fluids from an infected person enters the body of a person who is not infected

Prevention – Universal precautions

HIVAttacks Immune SystemInfects/Kills T-Helper Cells (CD4)Final Stage of HIV Infection: AIDS – Acquired

Immunodeficiency SyndromeOpportunistic infections – Kaposi’s sarcoma,

CMV Workers have been infected with HIV after

being stuck with needles containing HIV-infected blood or after infected blood gets into a worker’s open cut or a mucous membrane

Electron microscope image of HIV, seen as small spheres on the surface of white blood cells.

http://www.biologyimagelibrary.com/imagelibrary/images/12646_1_PHIL_1843_lores_BIL260805.JPG

HIVFragile retrovirusRisk of transmission after percutaneous exposure to

HIV infected blood is approximately .3% ROT after splash to eye/mouth/nose ~.1%57 healthcare workers with documented

occupationally acquired HIV infectionCDC ~56,300 new HIV infections in the

US in 2006

http://www.agen.ufl.edu/~chyn/age2062/lect/lect_14/Lect_14.htm

Healthcare Personnel with Documented and Possible Occupationally Acquired AIDS/HIV Infection, by Occupation, 1981-2006

Occupation Documented Possible

Nurse 24 35

Clinical Laboratory Worker

16 17

Nonsurgical Physician

6 12

Non-Clinical Laboratory Technician

3 -

Housekeeping or Maintenance

2 13

Dentist or Dental Worker

- 6

U.S. Infect Control Hosp Epidemiol 2003;24:86-96.

What is risk with lentiviral vectors?Potential for generation of replication-

competent lentivirus (RCL)Potential for insertional

mutagenesis/oncogenesisHost rangeMitigated by the nature of the vector system

(and its safety features) or exacerbated by the nature of the transgene insert encoded by the vector.

Universal PrecautionTreat all blood and body fluids as if infectious For labs: Biosafety Level 2 (BSL2)

containmentDefined by CDC/NIHCombination of laboratory practices and

techniques, safety equipment, and laboratory facilities

Many BSL2 safety procedures extend to non-lab settings

BSL2: Standard Microbiological PracticesAccess to lab limited during experimentsWashing hands after handling viable materials,

after removing gloves, and before leaving labNo eating, drinking, smoking, handling contacts,

or applying cosmeticsSafe handling of sharps, plastic substituted

for glassMinimize splash/aerosolsDecontaminate work surfaces daily and after spillsProper disposal of contaminated items as

biohazardous waste

Biohazard Waste DisposalGauzePlastic serological pipettesPlastic pipette tipsPetri dishesCulture vials/platesCounter top covers Contaminated PPE

SharpsContaminated broken glasswareScalpels, blades, capillary tubesSlides & cover slipsGlass pipettesGlass capillary tubesNeedles and syringes

Liquid WasteDecon. tissue and media with 10% bleach

solution~ 15 minute contactMay then be poured down drain USC does not have a permit to autoclave

medical waste!

BSL2: Special PracticesOnly those who have been advised of potential hazards may

enter labBiohazard sign when agent in use, include relevant contact

information, info on agent, what to do if exposed, PPEPI ensures all are appropriately trained on potential hazards of

work involved, how to prevent exposure, offered appropriate immunizations

Training on hazards, exposure prevention, SOP’sSharps precautions, engineered sharps, handling broken

glasswareInfectious agents placed in container with cover that prevents

leakage during collection, handling, processing, storage, transport, shipping

Decontamination of equipment/work surfaces Report exposures to PI, seek treatment

Safety engineered sharps2001 Needlestick Prevention Act

LabelingInternational biohazard symbolBiohazardous wasteDoor signsIncubatorsFreezersRefrigeratorsShipping boxes

DecontaminationWork surfaces, tools, and equipment1:10 bleach solutionLysol or other EPA registered disinfectant15 minute minimum decontamination time

Handling, storage, transport proceduresSpecimens must be placed in a container which

prevents leakage during collection, handling, processing, storage, transport, or shipping (use secondary container), container must be labeled with biohazard symbol

BSL2: Safety EquipmentRequires the use of a BSC for large volumes or potential

aerosol generationUse of centrifuge safety cups/sealed rotors for aerosol

containmentFace protection for anticipated splashes or sprays when

outside BSCLab coats, leave before leaving labGloves whenever potential contact with hazardous agents

Biosafety Cabinets

Biosafety CabinetsSweeping motions of armsDisinfect surfaces before and after useFront intakeUVBunsen burners alternatives (i.e. sterile

loops, touch-plate microburners)Substitute plasticware for glassware or

other sharp instruments whenever possible (i.e. pasteur pipettes)

Must be professionally decontaminated/recertified before moving

Recertify annually

BSL2: Lab facilities

Lockable doors Sink for handwashing No carpets/rugs Chairs covered in non-porous material BSC away from doors Eyewash station

Emergency procedures for spills Inform others in lab and restrict accessLeave area and allow aerosols to settleDon personal protective equipment If broken glass present, never handle directly by hand, use

forceps to remove and dispose into sharps containersPlace paper towels/absorbent over spillApply disinfectant(10% bleach) directly onto paper towels

and allow 15 minute contact time to disinfectClean spill by wiping around the perimeter and moving

inwardsDiscard materials as biohazardous wasteWash hands with soap and water

Emergency procedures for exposuresFlush affected area with water for 15 minutesInform supervisorKnow where to get treatment

Avoid working alone in lab!

Treatment LocationsHSC

Business HoursInternal Medicine (HCC)

1520 San Pablo St.Los Angeles, CA 90033

(323) 442-5100

After HoursWhite Memorial Hospital

1904 Bailey Street, Suite 100Los Angeles, CA 90033

(323) 222-9675

Exposure IncidentReport incident to supervisor immediately and

seek medical treatment! File Supervisor’s report of injuryFile Sharps Injury FormReport all exposure incidents to IBC

USC Exposure Control Plan

http://capsnet.usc.edu/LabSafety/BioSafety/BloodBornePathogensProgram

How to elim/minim exposure to human/NHP materials that might contain BBP

Outlines PI responsibility: Update ECP, GLS/BBP, Hep B vaccination

Methods of Compliance: Safety EquipmentGood Work Practices : PPE, hand washing,

labelingMedical Surveillance Program: PE eval./follow up

Contact InformationEnvironmental Health and Safety

323.442.2200(office)edavid@caps.usc.eduHSC, CHP 148