Bacteriology – Prelims PDF (KZNM BSMT - L2)

1 Bacteriology – Prelims KZNM BSMT – L2 LABORATORY SAFETY Medical Technologists / Microbiologists / Laboratory Staff should be KNOWLEDGEABLE, PROPERLY TRAINED & EQUIPPED with prop...

1 Bacteriology – Prelims KZNM BSMT – L2 LABORATORY SAFETY Medical Technologists / Microbiologists / Laboratory Staff should be KNOWLEDGEABLE, PROPERLY TRAINED & EQUIPPED with proper protective materials and working controls while working in the microbiology laboratory Chemical Safety All chemicals be identified Proper labeling is a must indicating health risks(carcinogen, mutagen, teratogen) and hazard class (corrosive, poison, flammable, oxidizing) CLASSES OF FIRE Hazardous chemicals must be inventoried CLASS A - Fires that involve solid or organic materials, annually such as wood, plastics, paper, textiles, or coal. All laboratories must have a file of every CLASS B - involve flammable liquids, such as gasoline, chemical they use and with corresponding petroleum oil, paint, or diesel MATERIAL SAFETY DATA SHEET (MSDS) CLASS C - involve flammable gases, such as propane, -MSDS includes: butane, or methane. o Substance name CLASS D - involve combustible metals, such as o Name, address and telephone number magnesium, lithium, sodium, potassium, titanium, or of manufacturer aluminum o Hazardous ingredient CLASS E - electrically energized equipment o Physical and chemical properties CLASS F - involve cooking oils and fats, such as o Fire and explosion data vegetable oil, sunflower oil, olive oil, maize oil, lard, or o Toxicity o Health effects and first aid butter (typically those used for deep-fat fryers) o Stability and reactivity o Shipping data Combination Type ABC extinguishers are found in o Spill, leak and disposal procedures most laboratories o Personal protective equipment Type C extinguisher containing CO2 or another o Handling and storage chemical to smother flames are also used (Does FUME HOODS are provided to prevent not damage equipment) inhalation of toxic fumes All sockets should be checked for electrical o Protection against chemical odor by grounding and leakage at least annually exhausting air to the outside No extension cords should be used in the laboratory Fire Safety Handling of Compressed Gases Fire evacuation plan is required Fire drills be conducted quarterly or annually Gas Cylinders (CO2, Anaerobic Gas mixture) depending on local laws contain pressurized gases and must be properly Exit paths must be clear of any obstructions handled and secured Fallen leaking cylinders o MISSILES (Loss of life and destruction to property) GAS TANKS o should be chained o Stored in well-ventilated area o Metal cap should always be in place o when tank is not in use o Should be transported chained in special dolly 2 Bacteriology – Prelims KZNM BSMT – L2 STOOL BIOSAFETY AND BIOSECURITY BODY FLUIDS Basic Concept of Biosafety & Biosecurity SPUTUM EXUDATES WORLD HEALTH ORGANIZATION ISOLATES Laboratory Biosafety Manual in 1983. (First Edition) Encouraged countries to accept and implement Laboratory Biosafety basic concepts in biological safety Achieved through: To develop national codes of practice for the safe o Various degrees of laboratory control and handling of pathogenic microorganisms in containment laboratories within their geographical borders. o Laboratory design and access restrictions Laboratory Biosafety Manual in 1993. (Second Edition) o Personnel expertise and training many countries have used the expert guidance o Use of containment equipment provided in the manual to develop such codes of o Safe methods of managing infectious practice materials Laboratory Biosafety Manual in 2004. (Third Edition) Addressing biological safety and security issues Laboratory Biosecurity facing us in the current millennium. Institutional and personal security measures Stresses the importance of personal responsibility. designed to prevent the loss, theft, misuse, New chapters have been added on risk diversion, or intentional release of pathogens and assessment, safe use of recombinant DNA toxins technology and transport of infectious materials. Achieved through: Recent world events have revealed new threats to o Facility security (limit access) public health through deliberate misuse and o Personnel reliability (background release of microbiological agents and toxins. o investigations) Introduces biosecurity concepts – the protection of o Information security (passwords) microbiological assets from theft, loss or diversion, o Transportation security (packaging) which could lead to the inappropriate use of these o Material accountability (inventory) agents to cause public health harm. Local Organizations on Biosafety Philippine Advanced Biosafety Officer Training (PhABOT) Expand the pool of competent biorisk officers and Biological Risk Association of the Philippines (BRAP), 2015 trainers in the Philippines A non-government and non-profit association work Strengthen the biorisk management system in the to serve the emergent concerns of biological risk Philippines management in various professional medical, agricultural, technological, and biological sectors Biohazards throughout the Philippines. Organisms infectious to humans Philippine Biosafety and Biosecurity Association, INC. Biologically active agents, animals or plants (PhBBA) causing disease in other living organisms leading to Created by a multi-disciplinary team, including significant impact to the environment or members from health care, academia, emergency community response, pharmaceutical/biotech, as well as from BACTERIA the executive, legislative and judicial branches of VIRUSES the government. FUNGI Fundamental Objective of Biosafety & PARASITES Biosecurity TOXINS ALLERGENS Containment of potentially harmful biological PRIONS agents. (Biosafety) VENOM “Keeping the people from bad bugs” RECOMBINANT DNA TECHNOLOGY Protection, control and accountability for valuable biological materials within laboratories, in order to Biological Agents prevent their unauthorized access, loss, theft, BLOOD 3 Bacteriology – Prelims KZNM BSMT – L2 misuse, diversion or intentional release. (Biosecurity) “Keeping the bad bugs from bad people.” Classification of Laboratory According to Biosafety Levels Biosafety Level 1 Appropriate for work with well-characterized, non- pathogenic organisms or agents (E. coli, B. subtilis) work is conducted in an open bench and no containment equipment (undergraduate & teaching laboratories) Use good laboratory practices, waste disposal, and aseptic techniques (clean work surfaces) & BIORISK MANAGEMENT handwashing AMP MODEL Biosafety Level 2 Appropriate for work with agents of moderate hazard associated with human disease by ingestion, percutaneous or mucus membrane exposure (Salmonella, HIV, HBV) Work has a restricted access & require a Assessment containment during certain processes (clinical a procedure that analyses and characterizes diagnostic laboratories) biological risks in a laboratory arising from a Recommended for laboratories handling human hazard or threat body fluids in which infectious agent maybe Biosafety Risk unknown o The risk of accidental exposure to or Biohazard warning sign posted in the lab. release of a biological hazard Biosafety Level 3 Biosecurity Risk o The risk of intentional removal (theft) of a Appropriate for work with exotic or indigenous valuable biological material agents with potential for aerosol transmission causing serious or lethal disease (Mtb, SARS) Effective treatment & vaccination available all work is contained, lab has engineering control to prevent release of the agent to the environment (eg. research & production facility laboratories) workers may need to wear a respirator Biosafety Level 4 Is reserved for work with dangerous agents that pose a high risk for life-threatening disease by infectious aerosols & from which no treatment or vaccine is available. (deadly viruses only-Ebola, Marburg, Lassa) Total containment, airtight labs, “submarine” doors, air pumps, water treatment, HEPA filtration & positive pressure “moonsuits” 4 Bacteriology – Prelims KZNM BSMT – L2 Mitigation Hierarchy of Controls Systematic approach to control, reduce, eliminate the risk of exposure from hazards. Steps in the hierarchy are in order of effectiveness Combination of 2 or more control measures to achieve an acceptable risk. Types of hazards, severity, exposure must be considered Elimination Or Substitution Removing the hazard, not working with the agent or replacing the hazard with something less dangerous Engineering Controls Physical changes to workstations, equipment, materials, production facilities, or any other relevant aspect of the work environment that reduce or prevent exposure to hazards A. LABORATORY ENVIRONMENT Air-handling system should move air from lower to higher risk areas, never the reverse Access should be limited to necessary personnel (biomedical engineers, janitors) Biohazard symbol must be prominently displayed on laboratory doors and any equipment (refrigerators, incubators, centrifuges) that contain infectious materials Visitors especially young children should be discouraged to enter Certain areas of high risk o (mycobacteriology and virology laboratories) should be closed to visitors B. BIOLOGIC SAFETY CABINETS (BSC) A device that encloses a workplace to protect workers from aerosol exposure to infectious agents 5 Bacteriology – Prelims KZNM BSMT – L2 Air containing infectious material is sterilized IIB either by heat, ultraviolet light, or most commonly IIB1 – 30% recirculated to the cabinet work area by passage to a HEPA filter through HEPA and 70% exhausted through a o High Efficiency Particulate Air (HEPA) filter dedicated duct to the outside through HEPA filter removes most particles larger than 0.3 um 100 fpm in diameter IIB2 – no recirculation, total exhaust to the outside B.1 CLASS I through a HEPA filter 100 fpm allow room air (unsterilized) to pass into the cabinet and around the material and area within, sterilizing only the air to be exhausted Have negative pressure Ventilated outside Operated with an open front 75 fpm B.1 CLASS II Sterilizes air that flows over the infectious material as well as air to be exhausted Class ll Type C1 Differentiated into Type A or B The type C1 cabinet is basically a type B1 cabinet IIA – mostly utilized in hospital settings in terms of airflow pattern. IIA1 – 70% recirculated to the cabinet work area The cabinet exhausts approximately 60% of the through HEPA and 30% exhausted through HEPA work zone airflow through a dedicated portion of back into the room or to outside through a canopy its centered depressed work tray/grill pattern and unit recirculates the remaining airflow, approximately 75 fpm 40% of work zone airflow through the non-portion IIA2 – similar to IIA1 but with 100 linear fpm uptake work tray grill area. under negative pressure to room What makes type C1 cabinet different from a type exhaust air can be ducted to outside through B1 is that it has an internal exhaust motor/blower canopy unit to push the airflow through the exhaust HEPA filter. Traditional type B1 cabinets require the facilities exhaust system to pull airflow through the exhaust HEPA filter and thus require to be direct connected. This new style cabinet is more like a type A2 with respect to exhaust in that it can be exhausted back into the room or through a canopy exhaust connection. In terms of exhaust requirements, the type C1 will use a bit more exhaust volume than a type B1 and a type B1 requires a bit more negative pull or static than a type C1. 6 Bacteriology – Prelims KZNM BSMT – L2 STANDARD PRECAUTIONS B.3 CLASS III In 1987 CDC published guidelines known as Completely enclosed UNIVERSAL PRECAUTION to reduce the risk of HBV Negative pressure transmission in clinical laboratories and blood Air coming into and going out of the cabinet is banks sterilized through double HEPA filter system In 1996 the safety recommendation became Cabinet is gas tight and sealed known as the STANDARD PRECAUTIONS. Operation performed through rubber gloves ALWAYS CONSIDER ANY SAMPLE AS POTENTIALLY attached to the cabinet INFECTIOUS Essentials of Standard Precautions and Safe Laboratory Work Practices: Extra belongings or bags must not be brought inside the microbiology laboratory Wear laboratory coats/gowns over street clothes Laboratory gowns should be removed before leaving the laboratory Do not eat, drink, smoke, or apply cosmetics (including lip balm) Do not insert or remove contact lenses Do not bite nails or chew on pens Administrative Controls Do not mouth-pipette Limit access to the laboratory to trained personnel Policies, standards and guidelines used to control only risks Assume all patients are infectious for all blood- Practices & Procedures borne pathogens Take special care to prevent injuries with sharp Processes and activities that have been shown in objects such as needles and scalp practice to be effective in reducing risks o Never recap needles by hand In the laboratory pathogens can become airborne o Practice fishing out its cap and securing it by: by pressing on the work surface o Removing evacuated tube caps o Discard sharps in an appropriate puncture o Manipulation of cultures resistant containers o Centrifuge, test tube cracks Report spills, breakage and injury Aerosol – Producing Devices o Spills should be contained by covering it o Centrifuge with absorbent material and saturate it o Needle with commercial germicide or 1:10 bleach o Syringes solution, allow to stand for 15 minutes o Pipettes before finally cleaning the surface. o Shakers Minimize aerosol production o Vacuum and Aspirating Equipments Wash hands before and after laboratory activity Disinfect laboratory benches/working surfaces before and after conduct of laboratory procedures Disinfectant (Household bleach – Sodium hypochlorite) must be diluted 1: 10 for porous or dirty surfaces and 1:100 for smooth surfaces and should not be more than 24 hours old. Identification and proper waste segregation Return all materials and reagents to proper storage area Handle equipments properly Keep the laboratory working surfaces free of clutter 7 Bacteriology – Prelims KZNM BSMT – L2 All materials contaminated with potentially Peel away from hand turning glove inside-out. infectious agents must be decontaminated before Hold in opposite gloved hand. disposal(chemical treatment, moist heat Slide ungloved finger under the wrist of the treatment, boiling) remaining glove, be careful not to touch the Infectious wastes must be placed in two –leak outside of the glove. proof plastic bags (double-bagging) Peel off from inside, creating a bag for both gloves Pipettes, swabs, and other glass objects should be Discard placed into rigid cardboard containers Proper washing - Don’t be a dope, wash with soap Broken glass is placed in thick boxes, when full Types of Gloves: must be autoclaved LATEX – most common type Sharp objects including scalpels and needles are o Made of natural rubber with outstanding placed in sharps containers tensile strength and temperature Avoid contaminating the benches, floors, walls and resistance wastebaskets o Resistant to most acids and bases Always use PERSONAL PROTECTIVE EQUIPMENTS o Available with or without powder (PPE) NITRILE – made of synthetic rubber o With high resistance to abrasions and Personal Protective Equipment punctures Devices worn by the worker to protect against o Especially suited for handling harsh hazards in the laboratory chemicals serves as protective barriers of the skin and o Substitute for latex type if allergic to latex mucosal surfaces against cuts, accidental needle NEOPRENE – made of synthetic rubber sticks, splashes and spills that might otherwise be o Resistant to most acids and bases, detrimental to the microbiologist / medical alcohols and solvents technologist o Remain flexible at low temperatures but Laboratory coats, gowns, aprons and coveralls are resistant to cuts, punctures and Laboratory coats and gowns are used to protect abrasions than are nitrile or rubber gloves from infectious fluids BUTYL – made of synthetic rubber Front button cotton laboratory coats may not be o High permeation resistance to vapors appropriate for working with large amount of o Ideal to use with ketones and esters infectious liquid Foot/Skin Protection Rear fastening Gowns may be appropriate for Open toed shoes, sandals and other open working at higher containment footwear should be prohibited Don’t wear laboratory coats outside of the lab or Any closed footwear will do to avoid injury to the take them home! feet during glass breakages, liquid spills Cuffed sleeves can protect the wrists and lower Disposable scrub shoes can be used arms Shorts and other garments that leave skin Gowns should not be laundered at home. unprotected are not appropriate GLOVES Open toed shoes, sandals and other open Wear disposable vinyl, synthetic or N-DEX nitrile footwear should be prohibited gloves when working with biohazardous materials Shorts and other garments that leave skin Avoid latex gloves (may cause allergies) unprotected are not appropriate Replace torn, soiled or damaged gloves Eye and Face Protection immediately PPE can protect mucous membranes and prevent Do not reuse gloves ingestion whenever there is potential for splash to Do not wear gloves outside of the laboratory eyes/face especially during the following: Wash hands after removing gloves o Spill Clean up Remove gloves prior to using “common” o Invasive procedures equipment or items that might be used by o Other high risk activities unprotected personnel Surgical masks with attached face shield protects Proper Glove Removal mouth, nose and eyes from droplets but does not Grasp outside edge near wrist. Careful not to touch protect from aerosols: It is not respiratory wrist with gloved hand protection 8 Bacteriology – Prelims KZNM BSMT – L2 Respiratory Protection Mailing Biohazardous Materials Designed as last resort or temporary control measure Respiratory protection program is 1. Must meet the requirements of International Air necessary to ensure safe and proper use Transport Association (IATA) and the International Two types: air supplying and air purifying Civil Aviation Organization (ICAO) Full face, half face, PAPR (Powered Air Purifying 2. Training in the proper packing and shipping is the Respirator) key feature of the regulation Special considerations: fit testing; facial hair; 3. Shipper is ultimately responsible for safe and comfort; care and maintenance appropriate packing Surgical masks are not respirators (look for the 4. Fines and penalties are responsibility of shipper N95) 5. Infectious specimens or isolates should be N95 respirators remove at least 95% of airborne wrapped with absorbent material and placed particles with a size of 300nm inside a plastic biohazard bag called PRIMARY N99 respirators remove at least 99% of airborne RECEPTACLE particles 6. Primary receptacle is then inserted into a N100 respirators remove at least 99.97% of secondary container, most often a watertight , airborne particles hard plastic mailer. HEAD CAPS 7. Secondary container is capped and placed inside Should hold hair securely especially for females an outer, tertiary container that protects it from with long hair physical and water damage 8. Coolant materials such as ice, dry ice, gel ice, liquid Females should securely tie their hair at the back nitrogen or cold dogs must be provided by the of their head and securely place the head cap shipper to ensure proper packaging 9. Forms necessary for the test is placed inside plastic wrapper and placed securely on top of the tertiary container 10. Shippers Declaration for Dangerous Goods Form must accompany Air Bill or Ground Form Performance The implementation of the entire biorisk Spill Management management system, including evaluating and ensuring that the system is working the way it was Spill Clean-up Materials designed. Disinfectant solution (10% freshly prepared bleach Another aspect of performance is the process of is effective in most cases) continually improving the system Forceps, tongs, broom, dust pan Risk Management Process Personal Protective Equipment (PPE): safety glasses, goggles, or face shield, utility gloves, wrap- around lab coat, shoe covers (optional) Biohazard' red bag, and sharps container if needed Paper towels, blue pads, or other absorbent Spill Involving Blood 1. Alert people in immediate area. 9 Bacteriology – Prelims KZNM BSMT – L2 2. Don PPE. Do not allow containers to overfill. 3. Cover an area twice the size of the spill with paper Never reach inside or attempt to force items into a towels. Pour disinfectant solution onto the spill, sharps container. starting at the perimeter and working inward from Discard all non-sharp material contaminated with the edges of the towels. Avoid splashing. blood, body fluids, or tissue into biohazard bags 4. Allow 20 - minute contact period. Keep an organized laboratory logbook and keep an 5. Wipe down any contaminated stationary inventory equipment or furniture with disinfectant. Entry to the lab limited to authorized personnel 6. Use forceps, tongs, or broom to remove broken only glass and other items; for glass, place in sharps Lock laboratory doors container or red bag if soft material. Wash hands after removing gloves, lab coats or 7. Remove towels and re-clean area with disinfectant other personal protective equipment and when solution. leaving the work area for general access areas such 8. Decontaminate (autoclave, chemical treatment) as lunch rooms, libraries, administration offices reusable clean-up items and other reusable Lab personnel having appropriate vaccinations equipment. prior to working with agent. 9. Inform personnel when the clean-up is complete Black bag (gen. Paper, Carton, Styropore, Boxes, 10. Make a report of the incident waste) Bottles, Syringe wrapper, Plastics, Good Laboratory Work Practices Diaper, Tin cans Green bag (wet Left over foods, Fruits & Vegetable A Good Laboratory Work Practice is a practice, waste) Peelings, Used cooking oil technique, or procedure that, when followed, has Yellow bag Used gauze, Catheter, Gloves, Plaster, been demonstrated to protect lab workers and the (infxs waste) Infectious diaper, Tubing's, Used environment and reduce the risk of exposure to cotton hazardous agents. Yellow bag w/ Empty bottles/containers of Beta dine, Perform procedures in a manner that will minimize black band Iodine, Acetone, Alcohol, Anaesthetic, splashing, spraying, spattering and generation of (chemical & Acids disinfectants, Laboratory droplets. If a procedure may cause aerosols or pharmaceutical Reagents, Expired and adulterated droplets to form, use containment such as a waste) drugs, Vials, Drugs for Chemotherapy, biological safety cabinet Used batteries Avoiding touching your skin or environmental Orange bag Radioactive Waste kept in leaded surfaces (e.g.; door knobs, phones, computer key (radioactive container (handled Separately) pads) or equipment while wearing gloves. waste) Iridium 192, Technetium 99, Iodine Decontaminate all work surfaces with a 131, Cobalt 90, Gallium 67 tuberculocidal disinfectant or sodium hypochlorite Sharps Needles, Syringes, Scalpels, Knives, solution (10% dilution of household bleach) collector Blades, Lancet, Any items that can following any spill and following completion of cause a cut or puncture wounds work. Use mechanical pipetting devices -never pipette by Barriers to Good Practices mouth. Convenience: Human blood, tissue and other potentially Practice: No food or drink allowed in the lab infectious materials should be transported in Problem: No lunch room capped containers which are placed in a second Result: Food stored and consumed in the lab leak proof container, appropriately labeled. Assumed Risk: Contamination, risk of infection, Never bend, break, recap, or otherwise manipulate accidental exposure needles. Don’t remove needles from syringe by Inventory: Practice: Update the inventory at the end of the hand. If removal is necessary, use a forceps, or day sharps containers equipped with a needle- Problem: It’s the end of the day, people are tired removing device on its lid. Result: Out of date or incorrect inventory that is Discard all sharps into approved sharps containers. retrospectively updated in the morning of several Sharps containers must be located in the days backlog immediate vicinity of sharps use. Assumed Risk: Theft, misuse, diversion, loss, Dispose of sharps containers when they are ¾ full. confusion 10 Bacteriology – Prelims KZNM BSMT – L2