Biological Safety Procedures PDF

MODULE 9: BIOLOGICAL SAFETY PROCEDURES Presented by: PMLS 1 Team of Instructors SPECIFIC LEARNING OUTCOMES At the end of the lesson, the students should be able to: 1. Explain accurately the significance of Occupational Safety and Health Administration (OSHA) in ensuring workplace safety, parti...

MODULE 9: BIOLOGICAL SAFETY PROCEDURES Presented by: PMLS 1 Team of Instructors SPECIFIC LEARNING OUTCOMES At the end of the lesson, the students should be able to: 1. Explain accurately the significance of Occupational Safety and Health Administration (OSHA) in ensuring workplace safety, particularly in the clinical laboratory. 2. Classify accurately various biological materials, such as blood, tissues, cells, and microorganisms, based on their characteristics and potential risks associated with handling each type. 3. Demonstrate effectively basic safety procedures for handling and transporting biological materials, including the use of personal protective equipment (PPE), proper labeling, secure packaging, and adherence to regulatory guidelines. SPECIFIC LEARNING OUTCOMES At the end of the lesson, the students should be able to: 4. Articulate clearly the importance of biological safety cabinets (BSCs) in protecting laboratory personnel, the environment, and the integrity of samples from biological hazards. 5. Demonstrate accurately detailed operating procedures and guidelines for biosafety, including the correct use of biosafety cabinets, decontamination methods, waste disposal, and emergency response protocols. 6. Explain correctly the appropriate placement and organization of materials and reagents within a biosafety cabinet to ensure optimal airflow, prevent cross-contamination, and maintain a sterile working environment. SPECIFIC LEARNING OUTCOMES At the end of the lesson, the students should be able to: 7. Describe correctly the importance of preventing biological contamination in the laboratory, explaining the potential consequences for patient samples, research outcomes, and laboratory safety. 8. Describe comprehensively the precautionary measures for preventing biological contamination, including proper aseptic techniques, regular cleaning and maintenance of equipment, and the correct use of PPE. 9. Explain clearly the relationship between the use of biosafety cabinets and the management of biological hazards and contamination, and how BSCs mitigate risks and protect both laboratory personnel and the environment. HISTORY OF BIOSAFETY FUNDAMENTAL CONCEPTS IN BIOSAFETY BIOLOGICAL MATERIALS SAFETY PROCEDURES IN HANDLING BIOLOGIC MATERIALS BIOSAFETY PROTECTS GERMS FROM PEOPLE.  BIOSAFETY ANSWER: PROTECTS PEOPLE FROM GERMS. BIOSECURITY PROTECTS GERMS FROM PEOPLE. ANSWER: WHEN DELAY IN TESTING OF URINE IS ANTICIPATED, THE BEST PRESERVATION METHOD IS REFRIGERATION (4°C)  EXCESS CSF SHOULD NOT BE DISCARDED AND SHOULD BE FROZEN UNTIL THERE IS NO FURTHER USE OF IT. ANSWER: IDEALLY, SPECIMENS SHOULD BE TRANSPORTED TO THE LABORATORY WITHIN 1 HOUR OF COLLECTION. THE IDEAL TRANSPORT ANSWER: TIME OF BIOLOGICAL SPECIMENS SHOULD BE WITHIN 2 HOURS OF COLLECTION ALTHOUGH THIS WILL DEPEND ON THE KIND OF TEST/S PERFORMED. ONE OF THE BASIS OF SPECIMEN REJECTION IS WHEN A SPECIMEN IS LEAKING. ANSWER: HISTORY OF BIOSAFETY Laboratory safety traces its history from US biological weapons program. The use of Class III safety cabinets and lamilar flows was evident during that time. Outside US, mechanical pipettors and ventilated cabinets were used. HISTORY OF BIOSAFETY In 1974, the CDC published the Classification of Etiological Agents on the Basis of Hazard. National Institutes of Health (NIH) published the NIH Guidelines for Reasearch Involving Recombinant DNA Molecules. HISTORY OF BIOSAFETY CDC and NIH jointly published the Biosafety in Microbiological and Biomedical Laboratories (1984). Biosafety officers emerged to adopt administrative roles. FUNDAMENTAL CONCEPTS IN BIOSAFETY Biosafety is the containment principles, technologies, and practices that are implemented to prevent unintentional exposure to pathogens and toxins, or their accidental release. “Biosafety protects people from germs.” Charles Baldwin (1966) created the biohazard symbol. It should be prominently displayed on laboratory doors and in any equipment that contain infectious material. CLASSIFICATION OF BIOLOGICAL MATERIALS BASIC SAFETY PROCEDURES IN HANDLING AND TRANSPORT OF BIOLOGICAL MATERIALS  COLLECTION A collection manual must have: Safety considerations Selection of the appropriate anatomic site and specimen Collection instruction Transportation instruction Labeling instruction Special instruction Sterile vs. Nonsterile collection Minimal acceptable quality and recommended quality COLLECTION Wala pa gali misulod ang dagom?! Agay ma’am kasakit! Collect before antibiotic therapy, ensure asceptic collection, quantity must be sufficient and must be placed in a sterile container COLLECTION COLLECTION TRANSPORT Specimens should be transported to the laboratory within 2 hours of collection. Containers should be leak-proof, sealed and has a separate section for paper work Bags should be labeled with biohazard symbol Consider the susceptibility of specimens to environmental conditions PRESERVATIVES Polyvinyl alcohol (PVA) and buffered formalin: for ova and parasite (O&P) Boric acid: microbial counts in urine Stuart’s Medium and Amie’s Medium: common holding / transport media 0.025% Sodium Polyanethol Sulfonate (SPS) : anticoagulant for blood culture Heparin: for viral cultures Ethylenediaminetetraacetic acid (EDTA): can be a viral PCR anticoagulant STORAGE 4°C (Refrigerator Temperature) 22°C(Ambient/Room Temperature) 37°C(Body Temperature) -20°C to - 70°C(Freezer Temperature) LABELING Specimens should be completely labeled with: Patient’s name Identifying number Birthdate Date and time of collection Source Initials of the individual that collected the sample REQUISITION A complete requisition includes: Patient’s name Hospital Identification number Age and date of birth Sex Collection date and time Ordering physician Exact nature and source of the specimen Diagnosis Current antimicrobial therapy REJECTION Unacceptable specimens include: Incomplete and incorrect label, and information that do not match with that of the request form Improper transport condition Quantity is insufficient Leaking specimen Specimen has dried KAMSAMII! REFERENCES: Baraclan-Orais, M. A. (NA.). Lecture handbook in PMLS 1 with science, technology and society. NA. Benitez, P. A. et al. (2019). Principles of medical laboratory science 1. NA. Quezon City: C&E Publishing, Inc. Delost, M.D.(2015). Introduction to diagnostic microbiology for the laboratory sciences. NA. Burlington, MA: Jones&Barlett Learning Forbes, Bailey and Scott. (2013). Diagnostic microbiology. 13th Ed. Singapore: Elsevier Lehman, D.C, & Mahon, C.R.(2019). Textbook of diagnostic microbiology. 6th Ed. St. Lupouis, Missouri: Elsevier Saunders Mcpherson, R.A. & Pincus, M.R. (2012). Henry’s clinical diagnosis and management by laboratory method. 22Ed. Philadelphia: Elsevier Strasinger, S.K. & Di Lorenzo M.S.(2011). The phlebotomy textbook. 3rd Ed. Philadelphia: F. A. Davis Company Strasinger, S.K. & Di Lorenzo M.S.(2014). Urinalysis and body fluids. 6th Ed. Philadelphia: F. A. Davis Company Lecture Notes in Microbiology 2021 by Sir Kristan: Rehistradong Medtech Online Review Tutorial Lecture Notes in Clinical Microscopy 2023 by Sir Kevin Aytona: Klubsybear Tutorials LESSON 10 1. Know the Importance of Biological Safety Cabinet 2. Discuss Significance of OSHA 3. Know the importance of Biological Contamination Biological Safety Cabinet OSHA Pathogen Regulations Biological Contamination A device that encloses a working area to protect workers from aerosol exposure and infectious disease agents. In BSC, the air contains the infectious material is sterilized, either by heat, UV light, or by passage through a high efficacy particulate air (HEPA) resistance Figure 1. Biological Safety Cabinet filter. CLASS I CLASS II CLASS III CABINET CABINET CABINET It is an open-fronted type of cabinet with negative pressure (ventilated cabinets). It allows room (unsterilized) air to enter the cabinet, cabinet, circulate around the area and exposure the material within; only the air to be exhausted is sterilized using HEPA filter. It is used for biosafety levels (BSL) 2 and 3 agents. also known as the laminar flow BSC commonly used BSC in a clinical microbiology laboratory (Class IIA). It sterilizes the air using HEPA filter that flows over the infectious material and the air to be exhausted used for BSL 2 and 3 agents There are two types of Class II Cabinets: Class IIA – has fixed openings; 70% of the air is recirculated Class IIB – variable sash opening; used for chemicals, radioisotopes, and carcinogens Most hospital clinical microbiology laboratory technologists use Class II BSC. MODEL STERILGARD SCHEMATIC SHOWING AIRFLOW It provides the highest level of safety to the worker. The air coming into and going out of the cabinets is sterilized using HEPA filter and the infectious material within is handled with rubber gloves that are attached and sealed to the cabinet. It is used for BSL 4 agents  SCHEMATIC SHOWING CLAS III SYSTEM AIRFLOW  Biosafety Level 1 Agents Biosafety Level 2 Agents (Based on Hazard) Biosafety Level 3 Agents Biosafety Level 4 Agents These are agents that have no known potential for infecting healthy people. This containment level is used in laboratory activities of students (for academic purposes). Some examples of pathogens that requires this containment level are Bacillus subtilis and Naegleria gruberi agents acquired by ingestion or exposure to percutaneous or mucous membrane include all the common agents of infectious diseases In handling these agents, access to the laboratory is limited. It also requires the personnel to change their clothes with the recommended laboratory clothing before going to their specific stations. The personnel handling these agents should also receive immunization. Some examples of pathogens that require this containment level are HIV, Bacillus anthracis, Yersinia pestis, Salmonella, and Shigella. These are potential agents for aerosol transmission In processing these lethal pathogens, the air movement in the laboratory must be controlled to contain the infectious materials. Some examples of pathogens that require this containment level are Mycobacterium tuberculosis, Francisella tularensis, Brucella spp., Coxiella burnetti, St. Louis encephalitis virus, and systemic fungi. These are agents that cause life-threatening infections In handling these organisms, maximum containment and decontamination of all personnel and materials before leaving the area are observed. Aerosol transmission with pressure is possible. Some examples of pathogens that require this containment level are arbovirus, arenavirus, filovirus, and smallpox virus. Universal Precaution a policy of treating all blood, tissue, body fluids and OPIM (other potentially infectious materials as infectious) Engineering and Work Practice Control involve taking physical steps to isolate or remove any possible pathogen hazards from the workplace Utilization of sharp’s containers, biological safety cabinet, laboratory fume foods, proper hand washing and hand washing facility, banning of eating, smoking and drinking inside the laboratory Personal Protective Equipment gloves must be worn when coming in contact with specimens and other hazardous chemicals and OPIM gloves must be replaced every after one patient and when the protective integrity is compromised Eye and face protection items Protective body clothing (laboratory gowns, aprons, laboratory coats, surgical caps, shoe covers and disposable arm sleeves) Housekeeping Techniques Must ensure that the workplace is maintained in a clean and sanitary condition Example: “Double-bagging” properly labeled with a warning and instructions for dealing a leak or spill generally, refers to contamination of our food or environment with microorganisms. - this means bacteria, viruses, fungi, and parasites. describes the event at which a foreign material or substance that can cause foodborne illnesses enters the food material. Contaminated sites have the potential to impact on human health and the environment in a variety of ways, depending upon type, concentration and locality of contamination, the exposure mechanism (e.g. ingestion, inhalation and so on) and the level of exposure.