Dr.Nagwa Aref

الحيوية المخاطر على التعرفالتحكم وطرق

الحيوية المخاطر على التعرفالتحكم وطرق

Biohazard Recognition and Control

?

Dr.Nagwa Aref

Dr.Nagwa Aref

Dr.Nagwa Aref

Dr.Nagwa Aref

المخاطر المخاطر بعض بعضسنة األمريكية 1941دراسة الواليات حصر في المتموجة 74تم الحمى بمرض إصابة حالة brucellosis ميكروب على أجريت

طريق عن إما العينات تلك مزارع مع التعامل عند والحذر الحيطة أخذ وعدم اإلهمال أو استالمها عند المرضى عيناتمع التعامل في المتاحة التقنية ضعف أو المالمسة أو العينات تلك من المتموجة الحمى بميكروب ملوث غبار استنشاق

االنتشار والسريع الخطير الميكروب هذا مثل

– منها 222وجود ذكرت 1949سنة وأبحاث تحاليل معامل بسبب كانت فيروسي بمرض إصابة قاتلة 21حالة كانت وكان حالةالسل مرض حاالت المصابة الحيوانات أو األنسجة مع السليم التعامل عدم بسبب معظمها ،

– الفنزويلي الدماغ التهاب مرض وباألخص والدماغ الكبد تليف والتهابات والتيفويد المتموجة، 20 الحمى % , (Venezuelan

encephalitis) المسبب أن فيعتقد الباقي أما اإلصابة إلى وأدت معروفة للميكروب التعرض طريقة فيها الحاالت تلك منالمباشر باالستنشاق التعرض األمراض كان لتلك المسببة المعدية المواد عينات مع المصابين تعامل بحكم (Arboviruses)

  عدة تضم المجموعة هذه و

– سنة عن تم 1967وفي لإلنسان 428الكشف تنتقل ممرضة فيروسات مجموعة مع التعامل بسبب بفيروسات إصابة حالةالحشرات طريق   عن

– (arthropod-borne viruses الصفراء الحمى فيروسات بأنواعها (yellow fever)  تشمل الدماغ التهاب المختلفة وفيروسات

 (encephalitis) للمعامل ، قادمة عينات من متطايرة Zجدا صغيرة بقايا أو غبار أو لبخار التعرض بسبب كان معظمها للتشخيص

– البحثية بالمختبرات العاملين من بشرية أخطاء بسبب معظمها وكانت دراسات عدة في ذكرت Zأيضا الفطرية اإلصاباتمثل ممرضة بفطريات الجلد عبر ملوثة مواد وحقن وخدش العيون على فطرية مزارع من قطرات تناثر Sporothrix) منها

schenckii) التنفسي الجهاز إصابات الطبي أو العفن تناثر بسبب

– (Blastomyces dermatitidis) و (Histoplasma capsulatum)

– وراء السبب كانت الميكروب الالمباالة من مستحلب ح_ضر عندما وذلك السحايا التهاب بمرض أشخاص خمسة إصابة

(Neisseria

– meningitidis) العزل حجرة المرض (safety cabinet)  خارج أعراض ظهور من أيام سبعة قبل ذلك وكانالطفيلية المعملية لLaboratory-acquired parasitic infections) واإلصابات

– النامية بالدول وخاصة ذلك من استثناء تكن م ،

– مناقشة تمت أخرى دراسة أدت 164وفي التي األسباب حيث من الطبية المختبرات بمجال للعاملين طفيلية إصابة حالة . طفيليات أمراض شملت إصابات ذلك إلى

Dr.Nagwa Aref

Dr.Nagwa Aref

Dr.Nagwa Aref

RISK LEVELS ASSOCIATED WITH

THE USE OF LABORATORY

ANIMALS

RECOMBINANT DNA AND GENETIC

MANIPULATION

CRITERIA FOR CLASSIFICATION OF

BIOLOGICAL AGENTS BY RISK GROUP

USE OF MAMMALIAN

CELLS IN CULTURE

Dr.Nagwa Aref

CRITERIA FOR CLASSIFICATION OF CRITERIA FOR CLASSIFICATION OF BIOLOGICAL AGENTS BY RISK GROUPBIOLOGICAL AGENTS BY RISK GROUP CRITERIA FOR CLASSIFICATION OF CRITERIA FOR CLASSIFICATION OF

BIOLOGICAL AGENTS BY RISK GROUPBIOLOGICAL AGENTS BY RISK GROUP

Risk Group

1

Risk Group

2

Risk Group

3

moderate individual risk,

limited community risk

high individual risk

low community risk

high individual risk

high community risk

Risk Group

4

low individual risk,

limited community risk

Dr.Nagwa Aref

Laboratory exposures rarely cause infection leading to serious disease

Effective treatment and preventive measures are available

The risk of spread is limited.

Risk Group 2

A pathogen that usually causes serious human or animal disease, or which can result in serious economic consequences

Does not ordinarily spread by casual contact from one individual to another

can be treated by antimicrobial or antiparasitic agents

Risk Group 3

A pathogen that usually causes serious human or animal disease,

can result in serious economic consequences but

does not ordinarily spread by casual contact from one individual to another, or that can be treated by antimicrobial or antiparasitic agents.

Risk Group 2

Dr.Nagwa Aref

BIOLOGICAL AGENTS BY RISK GROUPBIOLOGICAL AGENTS BY RISK GROUPBIOLOGICAL AGENTS BY RISK GROUPBIOLOGICAL AGENTS BY RISK GROUP

Risk Group 2Mycoplasma

Chlamydia

Bacteria

Parasites Viruses

Fungi

Agents listed in this group may be present in blood, CSF, central nervous system and other tissues ,

and infected arthropods, depending on the agent and the stage of infection .

parasites have caused laboratory infections by ingestion, skin or mucosal penetration or accidental injection .

Dr.Nagwa Aref

Bacteria Risk Group 2Bacteria Risk Group 2

Actinobacillus - all species

Actinomyces pyogenes (C. pyogenes)

Bacillus cereus

Bartonella bacilliformis, B. henselae, B. quintana, B.

elizabethae

Bordetella pertussis, B. parapertussis and B.

bronchiseptica

Borrelia recurrentis and B. burgdorferi

Campylobacter spp. (C. coli, C . fetus, C. jejuni)

Chlamydia pneumoniae, C. psittaci (non-avian strians),

C. trachomatis,

Clostridium botulinum, Cl. chauvoei, Cl. difficile, Cl.

haemolyticum,

Cl. histolyticum, Cl. novyi, Cl. perfringens, Cl. septicum,

Cl. sordellii, Cl. tetani

Corynebacterium diphtheriae, C. haemolyticum,

C. pseudotuberculosis, C. pyogenes (A. pyogenes)

Edwardsiella tarda

Erysipelothrix rusiopathae (insidiosa)

Escherichia coli enterotoxigenic/invasive/hemorrhagic

strains

Francisella tularensis Type B, (biovar palaearctica), F.

novocida

Fusobacterium necrophorum

Haemophilus influenzae, H. ducreyi

Helicobacter pylori

Legionella spp.

Leptospira interrogans - all serovars

Listeria monocytogenes

Mycobacteria - all species (except M. tuberculosis, and M. bovis

(non-BCG strain), which are in Risk Group 3)

Mycoplasma pneumoniae, M. hominis Neisseria gonorrhoeae, N.

meningitidis

Nocardia asteroides, N. brasiliensis

Pasteurella, all species (except P. multocida type B in Level 3)

Pseudomonas aeruginosa

Salmonella enterica (S. choleraesuis)

Salmonella enterica serovar arizonae (Arizona hinshawii)

Salmonella enterica ser. gallinarum-pullorum (S. gallinarum-

pullorum)

Salmonella enterica ser. meleagridis (S. meleagridis)

Salmonella enterica ser. paratyphi B (S. paratyphi B)

(Schottmulleri)

Salmonella enterica ser. typhi (S. typhi)

Salmonella enterica ser. typhimurium (S. typhimurium)

Shigella boydii, S. dysenteriae, S. flexneri, S. sonnei

Staphylococcus aureus

Streptobacillus moniliformis

Streptococcus spp. (Lancefield Groups A, B, C, D, G)

Treponema carateum, T. pallidum (including pertenue), T.

vincentii

Ureaplasma urealyticum

Vibrio cholerae (incl. El Tor), V. parahaemolyticus, V. vulnificus

Yersinia enterocolitica, Y. pseudotuberculosis

Dr.Nagwa Aref

– AdenoviridaeAdenoviruses, all serotypesArenaviridae

– Lymphocytic choriomeningitis virus (laboratory-adapted strains)

– Tacaribe virus complex: Tamiami, Tacaribe, PichindeBunyaviridae*

– Genus Bunyavirus

– Bunyamwera and related viruses

– California encephalitis group, including LaCrosse, Lumbo and snowshoe hareGenus Phlebovirus

– All species except Rift Valley fever virus Coronaviridae

– Human coronavirus, all strains

– Transmissible gastroenteritis virus of swine

– emagglutinating encephalomyelitis virus of swineGenus Lymphocryptovirus: Epstein Barr Virus (HHV 4) and EB-like isolates

– Genus Rhadinovirus: all isolates (except H. ateles and H. saimiri, see Risk Group 3)

– Genus Thetalymphocryptovirus: all isolatesUnassigned Herpesviruses: includes HHV 6 (human á-lymphotrophic virus), HHV 7, HHV 8, etc.Orthomyxoviridae

– Genus Influenzavirus:

– Influenza virus type A, all isolates

– Influenza virus type B, all isolates

– Influenza virus type C, all isolatesPapovaviridae

– Genus Papillomavirus: all isolates

– Genus Polyomavirus: all isolatesParamyxoviridae

– Mouse hepatitis virus– Bovine coronavirus– Feline infectious peritonitis virus– Avian infectious bronchitis virus– Canine, Rat and Rabbit coronaviruses

Flaviviridae*– Yellow fever virus (17D vaccine strain)– Dengue virus (serotypes 1,2,3,4)– Kunjin virus

Hepadnaviridae– Hepatitis B virus, includes Delta agen

Herpesviridae– Alphaherpesvirinae– Genus Simplexvirus: all isolates, including HHV 1 and HHV 2,

except Herpes B virus which is in Risk Group 4– Genus Varicellovirus: all isolates, including varicella/zoster virus

(HHV 3) and pseudorabies virus (see Table 1)Betaherpesvirinae

– Genus Cytomegalovirus: all isolates including CMV (HHV 5)– Genus Muromegalovirus: all isolates

GammaherpesvirinaeGenus Paramyxovirus: all isolates

– Genus Pneumovirus: all isolates

– Genus Morbillivirus: all isolates (except Rinderpest-see Table 1)ParvoviridaeGenus Parvovirus: all isolatesPicornaviridae

– Genus Aphthovirus - See Table 1

– Genus Cardiovirus - all isolates

– Genus Enterovirus - all isolates (see Table 1 for restrictions)

– Genus Hepatovirus - all isolates (Hepatitis A)

– Genus Rhinovirus - all isolates

– Poxviridae (see Table 1 for restrictions)

– Chordopoxvirinae (poxviruses of vertebrates)

– Genus Capripoxvirus

– Genus Molluscipoxvirus

– Genus Yatapoxvirus

– Genus Avipoxvirus - all isolates

– Genus Leporipoxvirus - all isolates

– Genus Orthopoxvirinae - all isolates (except Variola and Monkeypox in Level 4)

– Genus Parapoxvirus: all isolates

– vGenus Suipoxvirus: Swinepox (see Table 1 for restrictions)

– All other ungrouped poxviruses of vertebratesReoviridae

– Genus Orbivirus - all isolates (see Table 1 for restrictions)

Virus Risk Group 2

Dr.Nagwa Aref

BIOLOGICAL AGENTS BY RISK GROUPBIOLOGICAL AGENTS BY RISK GROUPBIOLOGICAL AGENTS BY RISK GROUPBIOLOGICAL AGENTS BY RISK GROUP

Risk Group 3

MycoplasmaChlamydia

Bacteria

Viruses

Fungi

Human immunodeficiency

viruses (HIV - all isolates

Bacillus anthracisBrucella - all species Burkolderia (Pseudomonas) mallei; B. pseudomalleiChlamydia psittaci - avian strains onlyCoxiella burnetiiFrancisella tularensis, type A (biovar tularensis)Mycobacterium tuberculosis; M. bovis (non-BCG strains)

Pasteurella multocida, type B

BunyaviridaeUnclassified Bunyavirus

Hantaan, Korean haemorrhagic fever and epidemic

nephrosis viruses including virus responsible for

Hantaviruspulmonary syndrome) Rift Valley fever virus

Flaviviridae*Yellow fever virus (Wild type)Japanese encephalitis virusMurray Valley encephalitis

virus

Dr.Nagwa Aref

BIOLOGICAL AGENTS BY RISK GROUPBIOLOGICAL AGENTS BY RISK GROUPBIOLOGICAL AGENTS BY RISK GROUPBIOLOGICAL AGENTS BY RISK GROUP

Risk Group 4

Viruses

Arenaviridae

Lassa, Junin, Machupo viruses, Sabia, GuanaritoBunyaviridae*

Genus Nairovirus

Crimean-Congo hemorrhagic feverFiloviridae

Marburg virus

Ebola virusFlaviviridae*

Tick-borne encephalitis complex, including -Russian Spring-Summer Encephalitis

Kyasanur forest virus

Omsk hemorrhagic fever virusHerpesviridae

Alphaherpesvirinae

Genus Simplexvirus: Herpes B virus (Monkey virus)Poxviridae

Genus Orthopoxvirinae

Variola

Monkeypox

Dr.Nagwa Aref

• Pathogenicity of material – disease incidence and severity

• Routes of Transmission – parenteral, airborne or ingestion

• Agent Stability – ease of decontamination • Infectious Dose – LD50 • Concentration – infectious organisms/vol. & working

volume • Origin of material - Wild Type, exotic, primary cells • Availability of effective prophylaxis – Hep. B vaccine • Medical surveillance – exposure management • Skill level of staff

Dr.Nagwa Aref

Risk AssessmentRisk Assessment

• Risk of Activity – same agent can have different containment levels at different stages of protocol: – Procedures that produce aerosols have higher risk – Procedures using needles or other sharps have higher risk – Handling blood, serum or tissue samples may have lower risk – Purified cultures or cell concentrates may have higher risk – Larger volumes (10 L) have higher risk

Dr.Nagwa Aref

Primary ContainmentPrimary Containment

• Lab practices – standard lab practice, limited access, biohazard warning sign, sharps/needle precautions, SOPs for decon, waste, medicals.

• Safety equipment – biosafety cabinets (BSC), sharps containers, sealed rotors.

• Personal protective equipment (PPE) – lab coat, gloves, goggles. • Host-vector for rDNA

Dr.Nagwa Aref

Dr.Nagwa Aref

Dr.Nagwa Aref

Four Levels of BiosafetyFour Levels of Biosafety

• BSL 1: Material not known to consistently cause disease in healthy

adults.

• BSL 2: Associated with human disease. Hazard is from percutaneous

injury, ingestion, or mucous membrane exposure.

• BSL 3: Indigenous or exotic agents with potential for aerosol

transmission; disease may have serious or lethal consequences.

• BSL 4: Dangerous/exotic agents which pose a high risk of life-

threatening disease, aerosol-transmitted lab infections or related

agents with unknown risk of transmission.

Dr.Nagwa Aref

Human Blood, Tissue and FluidHuman Blood, Tissue and Fluid

• Occupational Exposure to Bloodborne Pathogens • Occupational Safety & Health Administration (OSHA) 29 CFR

1910.1030

• Use BSL 2 work practices and procedures. • Additional requirements for HIV work. • Everyone needs to be offered the Hepatitis B vaccine. • Develop specific exposure plan SOPs. • Specific training is required. • Review needle/syringe use and replace with “safe” devices. • Exposure incidents must be followed up.

Dr.Nagwa Aref

Aerosol PrecautionsAerosol Precautions

• Use BSC for all procedures

that may generate aerosols. • Use centrifuges with

biosafety covers. • Do not use a syringe for

mixing infectious fluids. • Cultures, tissues, specimens

of body fluids, etc., are placed in a container with a cover that prevents leakage during collection, handling, processing, storage, transport or shipping.

Dr.Nagwa Aref

Select AgentsSelect Agents

• Possession, use and transfer of specific biological agents requires registration

• “Restricted Persons” are not allowed to have access to these agents.

• High security and containment must be maintained.

• Web site: http://www.cdc.gov/od/ohs/lrsat.htm

Dr.Nagwa Aref

• Control access to areas where biological agents or toxins are used and stored.

• Keep biological agents and toxins in locked containers.

• Know who is in the lab. • Know what materials are being brought into

the lab. • Know what materials are being removed from

the lab. • Have a protocol for reporting incidents. • Have an emergency plan.

Dr.Nagwa Aref

Waste DisposalWaste Disposal

• “Red bag” or “Regulated Medical Waste”: – All mammalian cells or anything that came in contact with mammalian

cells – All BSL 2 material or anything that came in contact with BSL 2 material – All needles/syringes regardless of use

• No need to autoclave this waste prior to disposal in EH&S red bag/box (material is incinerated).

Dr.Nagwa Aref

Dr.Nagwa Aref

المخلفات فرز نظام(Segregation System of )

المخلفات فرز نظام(Segregation System of )

المعدية المعدية الطبيةالطبيةالمخلفات المخلفات . 1 المعدية المعدية الطبيةالطبيةالمخلفات المخلفات . 1

البشرية . 2 واألنسجة األعضاء مثل األخرى المعدية الطبية البشرية المخلفات واألنسجة األعضاء مثل األخرى المعدية الطبية البشرية . 2 المخلفات واألنسجة األعضاء مثل األخرى المعدية الطبية البشرية المخلفات واألنسجة األعضاء مثل األخرى المعدية الطبية المخلفات

الحادة . 3 الحادة المخلفات الحادة . 3 المخلفات الحادة المخلفات المخلفات

والكيماوية . 4 الصيدالنية والكيماوية المخلفات الصيدالنية والكيماوية . 4 المخلفات الصيدالنية والكيماوية المخلفات الصيدالنية المخلفات

المشعة . 5 المشعة المخلفات المشعة . 5 المخلفات المشعة المخلفات المخلفات

الطبية . 6 الغير الطبية المخلفات الغير الطبية . 6 المخلفات الغير الطبية المخلفات الغير المخلفات

Dr.Nagwa Aref

العالمـــــــات العالمـــــــات الدولـــــــــيةالدولـــــــــية

العالمـــــــات العالمـــــــات المخلفات الطبية المشعةالدولـــــــــيةالدولـــــــــية

البيولوجية المخلفاتالمعدية والطبية

المخلفات الكيماوية السامة

مخلفات كيماوية حارقة

للمخلفات للمخلفات

البيولوجية البيولوجية

والطبيةوالطبية

للمخلفات للمخلفات

البيولوجية البيولوجية

والطبيةوالطبية

مخلفات سريعة االشتعالمخلفات سريعة االشتعال

مخلفات أدوية سامة

المخلفات الطبية ذات األضرار البيئية

المخلفات الكيماوية شديدة التفاعل مع الماء

Dr.Nagwa Aref

• OSHA Bloodborne Pathogens • http://www.osha.gov/SLTC/bloodbornepathogens/index.html • CDC Select Agents • http://www.cdc.gov/od/ohs/lrsat.htm • NIH Guidelines for Research Involving Recombinant DNA

Molecules • http://www4.od.nih.gov/oba/rac/guidelines/guidelines.html • DOT/CDC Shipping • http://www.cdc.gov/od/ohs/biosfty/shipregs.htm • CDC Import Permits • http://www.cdc.gov/od/ohs/biosfty/imprtper.htm • USDA/APHIS Permits • http://www.aphis.usda.gov/vs/ncie/

Dr.Nagwa Aref

• CDC: a permit is required to import etiologic agents of human disease and any materials, including live animals or insects, that may contain them. Unsterilized specimens of human and animal tissues (such as blood, body discharges, fluids, excretions or similar material) containing an infectious or etiologic agent require a permit in order to be imported.

• APHIS: a USDA veterinary permit is needed for materials derived from animals or exposed to animal-source materials.

Dr.Nagwa Aref

• Develop and practice plans for: • Spills: large spills, spills inside BSC

– Accidental exposures: needlesticks, eye/mucous membrane splash, breathing aerosols

– Power/Utility failures: BSC, freezers, ventilation, lights, water

– Fires – Medical emergencies

Dr.Nagwa Aref

ResourcesResourcesResourcesResources

• CDC Biosafety in Microbiological and Biomedical Laboratories

• http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4toc.htm

• ABSA Risk Groups • http://www.absa.org/

riskgroups/index.htm

• Canadian MSDSs • http://www.hc-

sc.gc.ca/pphb-dgspsp/msds-ftss/index.html

• Environmental Health & Safety – Lab Safety

• http://www.ehs.sunysb.edu or 2-9672