Safe Working Practices & Legislation (2020) - Student Notes PDF

Safe working practices and current legislation, including those relating to genetic manipulation Dr. Simon Gould  By the end of this lecture you should able; To understand the need for safety within the any laboratory (with a focus on micro). Identify the governing bodie...

Safe working practices and current legislation, including those relating to genetic manipulation Dr. Simon Gould  By the end of this lecture you should able; To understand the need for safety within the any laboratory (with a focus on micro). Identify the governing bodies and current legislation, including genetic manipulation. Description of the Hazard Group and the containment level required, plus identify micro- organisms in those groups. Appreciate the important and workload involved in laboratory safety. How many safety infringement can you spot Picture demonstrating three students no following safety regulation with a microbiology laboratory  Safe working practices “There are known knowns. These are things we know that we know. There are known unknowns. That is to say, there are things that we now know we don’t know. But there are also unknown unknowns. These are things we do not know we don’t know. ” -United States Secretary of Defence Donald Rumsfeld http://politicalhumor.about.com/cs/quotethis/a/ rumsfeldquotes.htm  Safe working practices #2 Why do we need them? – To reduce contact with sources of infection. – To avoid transmission of infection. – To reduce contact with sources of harmful chemical, radiation. Working with micro-organism involves: – “Good microbiological practice”. – Containment.  What are we trying to prevent ? Pictures showing laboratory injuries and new report Imperial collage London Foundation training in  Route of tranmission of microbes Ingestion Eating, drinking and smoking in the laboratory. Transfer of micro-organisms to mouth by: contaminated fingers or articles. Skin (percutaneous) Skin puncture by needle or other sharps. Bites and scratches by animals. Existing cuts or scratches. Conjunctiva Splashes of infectious material into the eye. Transfer to eye by contaminated fingers or articles. Inhalation Inhalation of airbourne hazards ie. aerosols. Pictures shown route of transmission https://www8.imperial.ac.uk/.../healthandsafety/Module %201%20Sept%2009v1.ppt  Good microbiological practice (GMP) & Containment What do we class as micro-organisms: – Bacteria – Fungi – Protozoa – Viruses – Prions  Good microbiological practice & Containment #2 When handling micro-organism aseptic techniques should be used at all times. Why? – To prevent contamination of the laboratory or personnel by the organisms being handled – To prevent contamination of the organisms from the environment or handler  Aseptic techniques include: – No mouth pipetting. – No eating, drinking, smoking or applying makeup or mobiles. – General tidiness and cleanliness: Cleaning up spillage with suitable agent. – Placing used pipette in disinfectant tip down. – Clearly label all specimens. – Loops sterilised immediately after use in Bunsen flame and ensuring the loop is not overload to prevent spitting. – Wash hand before leaving the laboratory. – No open toes or soft fabric shoes. – Designated Micro lab coat and safety glass  Aseptic techniques include: – Avoid creation of aerosols: o Don’t blow material from pipettes. o Don’t shake cultures too vigorously. o Don’t expose culture to air for too long. o Care should be taken with post-mortem dissection. o Care when grinding/homogenisation tissue. o Care when centrifugation – tube should be well sealed.  Current legislation Strict legislation for control of the handing of micro-organism. Control by a number of government bodies: – Health and Safety Executive (HSE) (Act 1974). – Advisory Committee on Dangerous Pathogens (ACDP). – The National Counter Terrorism Security Office (NaCTSO). These bodies have produced a number of reports – The Howie report – established code of practice for prevention of infection in clinical labs and post-mortem room. – Categorise pathogens according to hazard and category of containment (ACDP). – “The approved list of biological agent” last updated 2013  Current legislation #2 – Control of Substance Hazardous to Health (COSHH). – Genetic modified organism (GMO).  Safe in the laboratory: who’s job is it? The employer: – Must provide safety training and equipment fit for purpose. – Biological Safety Office (BSO) who sits on the safety committee: – Hazard Advise on microbiological safety. – Hazard groups that can be safely work with: – Containment. – Disinfection. – Safe dispose of waste material. The employee “YOU AS A WORKER”. – Legally obligation.  Assessment of risk Microbial pathogens are categorised according to hazard and containment by ACDP. Micro organisms are categorised in one of four group: Hazard group 1 contains micro-organisms with the lowest level of risk. Hazard group 4 contain micro-organisms with the highest level of risk.  Containment Hazards groups handle in same level containment (laboratory). Hazard Hazard Hazard Hazard Group 1 Group 2 Group 3 Group 4 Increasing hazard to human health Lab containment Lab containment Lab containment Lab containment Level 1 Level 2 Level 3 Level 4 https://www8.imperial.ac.uk/.../healthandsafety/Module %201%20Sept%2009v1.ppt  Hazard Grouping (HG) Example of Hazard group 1 pathogens: – Bacteria: Bacillius subtilis, Lactobacillus casei – Yeast: Saccharomyces cerevisiae Example of hazard group 2 microbes: – Bacteria: Escherichia coli, methicillin resistant Staphlyococcus aureus, Clostridium tetani (tetanus). – Fungi: Candida albicans (thrush), Aspergillus sp. (lung disease). – Parasite: Toxoplasma gondii (toxoplasmosis), Schistosoma mansoni (schistosomiesis). – Virus: Epstein-barr virus (glandular fever), Measles virus, Mumps virus, polio virus (vaccines existed), BCG vaccine. http://www.hse.gov.uk/biosafety/biologagents.pdf  Hazard Grouping #4 Example of hazard group 3 microbes: – Bacteria: Bacillus anthrasis (anthrax), Mycobacterium tuberculosis (tuberculosis). – Fungi: Histoplasma (histoplasmosis). – Parasite: Trypanosoma brucei rhodesiense (toxoplasmosis). – Viruses: HIV, Rabies virus, Hepatitis. http://www.hse.gov.uk/biosafety/biologagents.pdf  Hazard Grouping #6 Examples of hazard group 4 microbes: – Only consists of viruses: Lassa fever, Congo Hemorrhagic fever virus, Ebola fever, Herpes virus simiae, Smallpox virus (white pox virus) http://www.hse.gov.uk/biosafety/biologagents.pdf  Category 4 laboratory #2 Secondary control measure The worker Primary control measure Figure showing the separation of areas in CAT 4 lab Bio-hazard suit  Microbiological safety cabinets (MSC) Required for work at containment level 2 and 3 if the work presents a risk of aerosol exposure: Must be correctly selected (type and make). Must be correctly installed. Must be correctly used. Must be correctly maintained. https://www8.imperial.ac.uk/.../healthandsafety/Module %201%20Sept%2009v1.ppt  MSC - class I High Efficiency Particulate Air (HEPA) filter USER Figure showing the air flow into the cabinet  MSC - class II High Efficiency Particulate Air (HEPA) filter Downflow HEPA filter USER Figure showing the air flow into the cabinet https://www8.imperial.ac.uk/.../healthandsafety/Module %201%20Sept%2009v1.ppt  MSC – class III HEPA filter HEPA filter Figure showing the air flow into the USER cabinet https://www8.imperial.ac.uk/.../healthandsafety/Module%201%20Sept%2009v1.ppt http://www.google.co.uk/imgres?imgurl=http://web.princeton.edu/sites/ehs/biosafety/livevirusworker/18a-Autoclavetape.jpg&imgrefurl=http://web.princeton.edu/sites/ehs/biosafety/livevirusworker/decontamination.htm&usg=__mazENUD732mS5lXuhnwOaDsszho=&h=960&w=1280&sz=82&hl=en&start=47&zoom=1&itbs=1&tbnid=NMc-zURl3JWT- M:&tbnh=113&tbnw=150&prev=/search%3Fq%3Dtaping%2Bup%2Ba%2Bautoclave%2Bbags%26start%3D40%26hl%3Den%26sa%3DN%26ndsp%3D20%26biw%3D1259%26bih%3D878%26tbm%3Disch%26prmd%3Divns&ei=AbbLTfa-LMGGswbntbXbAw  MSC – airflow sensitivity MSC bench Do not have another worker at an adjacent bench MSC 300 mm bench Allow adequate room for workers at nearby benches https://www8.imperial.ac.uk/.../healthandsafety/Module %201%20Sept%2009v1.ppt  Movement of microbes Move of pathogen between laboratory has always been problematic. – Regulation have been tighten since early 2000. Regulated by number of agencies: – Department of transport Transport of infectious substance (2006). – ACDP, NaCTSO and HSE. Hazard group 3 and 4 microbes must not sent through the post. – Group 4 microbes usually have a police escort and expert used to handling the pathogen. – Group 4 microbes also require government approval before movement. http://webarchive.nationalarchives.gov.uk/+/http://www.dft.gov.uk/pgr/freight/dgt1/guidance/guidancenonclass7/guidanceontransportofinfecti3186  Movement of microbes cont. Specimen should be in: Secondly container – In a double container – Container should be metal/plastic with a screw top – Labelled clearly with biohazard warning Primary container  Genetic manipulation/modification organism (GMO) Genetic make up of microbes may change naturally (antibiotic resistance) or intentionally in the laboratory. – Therefore may be necessary to up-grade their hazard group. Regulated by number of agencies: – Guidance from the Scientific Advisory Committee on Genetic Modification (SACGM); HSE. ADCP. Department for Environment, Food and Rural Affairs (DEFRA). Scottish Executive Environment and Rural Affairs Department (SSREAD).  GMO #2 Within laboratory GMO is regulated by: – GM committee and the BSO. – Are subject to all regulations that affect non GMO microbiology laboratories. – Subject to a higher level of monitoring Health surveillance of employees to detect adverse reaction/ill health to prevent further exposure. – Choose the less hazardous microbe to manipulate.  GMO #3 GM risk assessment should consider the potential impact to: – Human health. – Animal health. – The environment. GM risk assessment should be reviewed regularly and up date in light of new knowledge: – These risk assessments must be kept for 10 years after the work has end.  Summary Safe working practises in any laboratory is paramount. – Especially important in microbiology when the risk may be unknown. It is a highly legislated by a number of bodies: – HSE, ACDP, NaCTSO, DoT, DEFRA. Micro organism are categorised into group with HG1 being the lowest group and 4 being the highest group. HG are manipulated at the same containment level.  Future reading Biological agents: Managing the risks in laboratories and healthcare premises – http://www.hse.gov.uk/biosafety/biologagents.pdf The approved list of biological agents – http://www.hse.gov.uk/pubns/misc208.pdf 32