Laboratory Biosafety andBiosecurity Issues Relatedto Brucella Research andDiagnosticsJoseph P. Kozlovac, M.S., RBP, CBSPAg...
Brucellosis: Who is at Risk?• Farmers• Shepherds, GoatHerders, Cattlemen• Veterinarians• Laboratory Workers• Abattoir Work...
Routes of Exposure• Human infectious dose10-100 organisms byrespiratory route• Mucous membrane,conjunctivae or non-intacts...
•First documented LAI with Brucellosis (Brucellamelitensis) occurred in 1897 via syringe•1930-1978, 426 LAIs, 5 Deaths (Pi...
Biosafety Risk Assessment:Safety Risk Groups• Risk Group 1– No or low individual and community risk– Unlikely to cause hum...
BSL for Diagnostic Work• BSL-2 practices,containment equipmentand facilities.• Operations thatgenerate aerosols orsplashes...
BSL-3 for CultureandAnimal Studies• Biosafety level 3 isappropriate for handlingBrucella cultures orinfected membranes,fet...
8Lab Procedures That Emit Aerosols• Pipetting• Mixing• Shaking• Centrifugation• Grinding• Blending• Vortexing• Sonic Disru...
Engineering Controls• Primary Barriers– Biological SafetyCabinets (BSC)– Enclosed Containers– Animal Isolators
10Personal Protective Equipment• Gloves• Coats• Gowns• ShoeCovers• Boots• Respirators• Face Shields• Safety Glasses• Safet...
Occupational Health andMedical Surveillance• All persons working with virulent Brucellae should bekept under close clinica...
Occupational Health andMedical Surveillance• Emergency Response or Treatment– Laboratory exposure– Accident or injury in t...
• Program to prevent loss, theft or misuse ofmicroorganisms, biological materials, and research-related information– Prote...
COMPONENTS OFLABORATORY BIOSECURITY
Basic Truism About Security SystemsA security system cannot protect every asset against everyconceivable threat• Security ...
The Take Home Message• Comprehensive biosafety/biosecurity trainingand consistent procedures for all personnelworking with...
Laboratory biosafety and biosecurity issues related to Brucella research and diagnostics
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Laboratory biosafety and biosecurity issues related to Brucella research and diagnostics

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Presented by Joseph P. Kozlovac at a workshop on an integrated approach to controlling brucellosis in Africa, Addis Ababa, Ethiopia, 29-31 January 2013.

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  • The risk of exposure and LAI in laboratory workers tends to be lower as compared to other health care workers, however infection of clinical and research laboratory workers is greater than the general public.
  • The 1 st case documented by Kisskalt in 1929. 1930-78 Brucella was the #1 reported LAI and 1979-2004 Brucella was #4. The 4 deaths were all aborted fetuses Brucellosis had long been recognized as a dangerous lab agent. During winter of 1938-1939 94 lab acquired cases due to generation of aerosols from a centrifugation operation mostly among students in a 3 story building. This case was followed by a survey conducted by Meyer and Eddie in 1941. They reported on 76 brucella infections beginning in 1897 and on 74 lab infections in the U.S. between 1922-1939. Brucellosis is in fact one of the most easily acquired laboratory infections. The degree of risk varies, not only with the virulence of the organism, B. melitensis and B. suis being the most dangerous for humans, but also with the numbers of bacteria in the material being handled. Picture. Brucella Militensis gram stain from culture
  • BSL-2 practies, containment equipment, and facilities are recommended for routine clinical specimens of human or animal origin. Products of coneption containing or believed to contain pathogenic Brucella due to the high concentrations (10 9 organisms per gram of tissue) it is recommend to minimally use BSL-3 practices with products of conception Blood samples and biopsy material for either serological or bacteriological diagnosis will rarely contain Brucellae in sufficient numbers to present a significant risk to personnel handling them but should still be handled with care at Biosafety level 2. Normally these will be dealt with in general diagnostic sections along with samples that may contain other human pathogens.
  • Sniffing of bacteriologic cultures and conducting operations that generate aerosols have resulted in LAIs
  • Eppendorf Centrifuge
  • Photos Ward 200: The Slammer, USAMRIID’s BSL-4 Patient Isolation Ward Persons at Risk Exposure Activities PEP Recommendations Follow up/Monitoring Person performing activity and any person within a 5 ft. radius Work with a Brucella isolate Sniffed or opened culture plate Mouth pipetted specimen material Worked in Class II biosafety cabinet or on open bench without using BSL-3 precautions Doxycycline 100mg twice daily and rifampin 600mg once daily for 3 weeks TMP-SMZ should be considered for patients with contraindications to doxycycline Persons with contraindications to rifampin should consult with their HCP Pregnant women should consult with obstetrician Sequential serologic testing at 0, 6, 12, 18 and 24 weeks post exposure Symptom watch (e.g. weekly) and daily self fever check for 24 weeks No serological monitoring available for RB51 and B. canis exposures
  • The term “biosecurity” has multiple definitions. In the animal industry, the term biosecurity relates to the protection of an animal colony from microbial contamination. In some countries, the term biosecurity is used in place of the term biosafety. For the purposes of this chapter the term “biosecurity” will refer to the protection of microbial agents from loss, theft, diversion or intentional misuse. This is consistent with current WHO and American Biological Safety Association (ABSA) usage of this term.
  • Restrict Access Key or Card reader entry Biometric control Only individuals that are properly trained and have a NEED to be in the laboratory should be granted access
  • Laboratory biosafety and biosecurity issues related to Brucella research and diagnostics

    1. 1. Laboratory Biosafety andBiosecurity Issues Relatedto Brucella Research andDiagnosticsJoseph P. Kozlovac, M.S., RBP, CBSPAgency Biological Safety OfficerUSDA Agricultural Research ServiceBeltsville, MD 20705-5146Workshop: An Integrated Approach to Controlling Brucellosis in Africa, AddisAbaba, 29-31 January 2013
    2. 2. Brucellosis: Who is at Risk?• Farmers• Shepherds, GoatHerders, Cattlemen• Veterinarians• Laboratory Workers• Abattoir WorkersOccupational Risk Non-Occupational Risk• Travellers• Hunters• Consumers ofunpasteurized dairyproductsInteresting Factoid: Over the past ~60 years2% of all brucellosis cases have beenlaboratory acquired.Yagupsky, P. and Baron, E.J. Laboratory Exposures to Brucellae and Implicationsfor Bioterrorism, Emerging Infectious Diseases . 2005 . 11(8) 1180-1185
    3. 3. Routes of Exposure• Human infectious dose10-100 organisms byrespiratory route• Mucous membrane,conjunctivae or non-intactskin contact with culturesor infected tissues• Ingestion (mouthpipetting)• Percutaneous– Including accidentalinoculation with vaccinestrains• B. abortus stain 19, RB51• B. militensis Rev-1• Person to person transmissionrare• Brucella antigenhypersensitivity Rxn is rarehazard in lab staff andoccasional hazard in animalcare staff
    4. 4. •First documented LAI with Brucellosis (Brucellamelitensis) occurred in 1897 via syringe•1930-1978, 426 LAIs, 5 Deaths (Pike 1978)•1941, Meyer and Eddie reported 76 Brucella LAIsbeginning in 1897 and 74 lab infections in the UnitedStates between 1922-1939•1979 -2004, 143 LAIs, 4* Deaths (Beyers 2006)Brucellosis: Historical RecordedLaboratory – Associated Infections
    5. 5. Biosafety Risk Assessment:Safety Risk Groups• Risk Group 1– No or low individual and community risk– Unlikely to cause human or animal disease• Risk Group 2– Moderate individual risk, low community risk– Can cause disease but unlikely to be a serious hazard. Labexposures may cause serious infection, but effective treatment andpreventative measures are available and risk of spread of infectionis limited.• Risk Group 3– High individual risk, low community risk– Usually causes serious human or animal disease but does notordinarily spread. Effective treatment and preventativemeasures are available.• Risk Group 4– High individual and community risk– Usually causes serious human or animal disease and can bereadily transmitted. Effective treatment and preventative measuresare not usually available.
    6. 6. BSL for Diagnostic Work• BSL-2 practices,containment equipmentand facilities.• Operations thatgenerate aerosols orsplashes should beconducted within aBiological SafetyCabinetBSL-2
    7. 7. BSL-3 for CultureandAnimal Studies• Biosafety level 3 isappropriate for handlingBrucella cultures orinfected membranes,fetal tissues and fluids.• ABSL-3 practices,containment equipmentand facilities arerecommended for animalstudiesBSL-3
    8. 8. 8Lab Procedures That Emit Aerosols• Pipetting• Mixing• Shaking• Centrifugation• Grinding• Blending• Vortexing• Sonic Disruption• Opening LypholizedCultures• Flaming bacteriologic loops• Entering or openingvessels to sample liquidunder pressure
    9. 9. Engineering Controls• Primary Barriers– Biological SafetyCabinets (BSC)– Enclosed Containers– Animal Isolators
    10. 10. 10Personal Protective Equipment• Gloves• Coats• Gowns• ShoeCovers• Boots• Respirators• Face Shields• Safety Glasses• Safety Goggles• Hearing Protection
    11. 11. Occupational Health andMedical Surveillance• All persons working with virulent Brucellae should bekept under close clinical and serological surveillance• Educate staff on human symptoms and have policy inplace for staff to report symptoms• Consider providing laboratory workers with medicalcards which include, at a minimum, the followinginformation:− Genus and species of organism which they work with− 24-hour contact numbers for principal investigator andinstitution’s occupational health care provider(s)
    12. 12. Occupational Health andMedical Surveillance• Emergency Response or Treatment– Laboratory exposure– Accident or injury in the laboratory– Post-exposure samples (recommended sequential serologictesting at 0, 6, 12, 18, and 24 weeks post exposure)– Provide treatment (Doxycycline 100mg twice daily andrifampin 600mg once daily for 3 weeks)– Re-examination of the patient before allowing them to returnto work after a laboratory exposurehttp://www.cdc.gov/brucellosis/laboratories/risk-level.html
    13. 13. • Program to prevent loss, theft or misuse ofmicroorganisms, biological materials, and research-related information– Protect pathogens from dangerous people– Limit access to areas that contain certain biologicalagents or assets• Guidance documents– WHO Laboratory Biosecurity Guidance, 2006http://www.who.int/csr/resources/publications/biosafety/WHO_CDS_EPR_2006_6.pdf– U.S. BMBL Section VI—Principles of Laboratory Biosecurityhttp://www.cdc.gov/biosafety/publications/bmbl5/index.htmDefining Laboratory Biosecurity
    14. 14. COMPONENTS OFLABORATORY BIOSECURITY
    15. 15. Basic Truism About Security SystemsA security system cannot protect every asset against everyconceivable threat• Security resources are not infinite• Security systems should be based on the asset or materialthat requires protection• Security systems should be designed to address uniqueoperations
    16. 16. The Take Home Message• Comprehensive biosafety/biosecurity trainingand consistent procedures for all personnelworking with Brucella species is needed• Brucellosis has been documented as one of themost frequently acquired laboratory infections.the importance of using appropriate biosafetypractices and facilities cannot be overemphasized.• Laboratory security measures should be basedon a sound risk assessment but should notnegatively impact the research mission.

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