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DRAFT - under revision

12. BIOLOGICAL HAZARDS

General

WHAT IS REGARDED AS A BIOLOGICAL MATERIAL?
  • All Biological tissues and fluids including blood and Urine.
  • All biological materials MUST be treated as having the potential to cause harm/infection.
  • A risk assessment MUST be undertaken before comencing any work that involves handling biological tissues or fluids.
  • If Appropriate, a COSHH assessment should also be carried out before work Starts.
  • It is imperative that all personnel who deal with biological experimental procedures should be conscious of the possible hazards that may arise. If in doubt you must consult your veterinary or medical/clinical partners.

    Good Working Practices

    If human tissues of any kind are being used, the individual using them MUST be immunized against hepatitis B. This should be arranged either at the Student Health Centre or at the Individual's General Practitioner.

      HANDLING BIOLOGICAL TISSUES - GENERAL Laboratory behaviour
      • Laboratory coats MUST be worn in the laboratory and removed when leaving the laboratory.
      • Personal protective equipment, including protective clothing, must be:
        • stored in a well-defined place;
        • checked and cleaned at suitable intervals;
        • when discovered to be defective, repaired or replaced before further use.
        • To Minimise risk of injury STURDY footwear MUST be worn when performing disections or using surgical blades.
      • Personal protective equipment which may be contaminated by biological agents must be:
        • removed on leaving the working area;
        • kept apart from uncontaminated clothing;
        • decontaminated and cleaned or, if necessary, destroyed.
      • Where ever possible all human/animal tissues/fluids should be contained and handled within a spill tray.
      • All procedures should be performed so as to minimise the production of aerosols, this is especially important when undertaking procedures such as sawing bone.
      • Eating, chewing, drinking, taking medication, smoking, storing food and applying cosmetics is prohibited in laboratories.
      • Persons working in biological laboratories should adhere to a strict code of personal hygiene
      • Hands must be decontaminated immediately when contamination is suspected, after handling infective materials and before leaving the laboratory. When gloves are worn, these should be washed or preferably changed before handling items likely to be touched by others not wearing gloves, for example, telephones, paperwork, computer keyboards and, where practicable, equipment controls should be protected by a removable flexible cover that can be disinfected.
      • Mouth pipetting is forbidden.
      • Bench tops and work surfaces should be regularly decontaminated according to the pattern of work and should be cleaned and disinfected after use.
      • Used glassware and other materials awaiting disinfection should be stored in a safe manner. Pipettes, for example, if placed in disinfectant, should be totally immersed.
      • Currently there are no autoclaving facilities within the college as the nature of work currently being undertaken does not require this. It is therefore essential that adequate disinfection takes place
      • There must be safe storage of biological materials.
      • All waste material, if not to be incinerated, should be disposed of safely by other appropriate means.
      • Accidents and incidents MUST be immediately reported to and recorded by the person responsible for the work or other delegated person.
      • Effective disinfectants should be available for immediate use in the event of spillage.
      • Minimum paper only should be in laboratories ie essential paperwork and laboratory books only.
      • All potentially infectious and toxic materials MUSTbe correctly labelled and storage areas should show appropriate warning notices.
      • If laboratories are handing biological materials which could cause illness they MUST have a system whereby illness is reported without delay.
      • Great care should be exercised in using hypodermic syringes. Apart from the risk of accidental self inoculations, spraying may occur if a needle becomes loosened from the syringe during an injection.
      • Laboratory personnel must receive suitable and sufficient information, instruction and training in the procedures to be conducted in the laboratory.
      • Training received must be recorded on a laboratory proforma and signed by the trainee. This should also include details of relevant documents read.
      • Every researcher must have a nominated academic supervisor responsible for ensuring compliance with these rules.
      • Individuals working with biological materials must ensure that other persons eg cleaners, maintenance personnel, visitors etc are not exposed to biological hazards.
      • Cleaning staff must only deal with general waste and do not handle waste for incineration (yellow bag waste), biohazardous or other similar waste. This MUST be dealt with (including transportation to the wheelie bins) by lab workers.
      • Project students and other undergraduates working with biological materials require close supervision, the safety and security of these individuals is paramount

      DISPOSAL OF SHARPS

    • Sharps include items such as scalpal blades, needles, broken glass, ampoules and pasteur pipettes
    • NO sharps should be left lying around on the open bench as this poses a real risk on injury/infection to the individaul and other staff/students
    • ALL sharps MUST be disgarded IMMEDIATELY after use in designated yellow sharps bins
    • NEVER, EVER attempt to recover objects from the sharps bin
    • DO NOT over fill sharps bins, seal full bins by sliding the lid across FULLY and placing in a biohazard bag for disposal by incineration
    • THE HANDLING AND CENTRIFUGATION OF BLOOD:

      A risk and COSHH assessment should be completed before any work using blood samples proceeds. The risk assessment should be specific to the work being performed and it is essential to state the source of the blood.

      There is a risk of transmitting blood borne viruses, therefore, ALL blood samples must be treated as being potenially infectious. Infection can be passsed through skin and mucous membrane.

      ALL individuals handling human blood MUST be immunized against Hepatitis B.

      If at all possible blood should be screened (i.e. out-of-date or excess blood for the National Blood Bank). In some cases an individual may use their own blood for their research. With this exception unscreened blood MUST NOT be used.

      Blood must always be contained in leak-proof containers and in sealable buckets.

      Protective Colthing Lab coats, gloves and eye protection should be worn to protect from spills and splastes of blood. It is advisable to wear a plastic apron in addition where large quantities of blood are being used.

      Before starting work the individual MUST ensure that there is a blood spill kit is at hand.

      As much of the process as possible (e.g. transferring blood between containers), should be performed in a spill tray

      CENTRIFUGING BLOOD:

      It is essential that safe procedures are adheared to at all times when using centrifuges, this is of particular importance when handling human blood or bloody fluids:

    • Always spin blood in centrifuge tubes with well fitting lids.
    • When spinning blood only use centrifuge buckets with lids that form a seal.
    • Ensure centrifuge tubes are the correct size for the rotor.
    • DO NOT over fill centrifuge tubes.
    • Ensure the tubes are correctly balanced.
    • DO NOT exceed maximum sttings for particular centrifuge and rotor.
    • After use wipe around centrifuge bowl with antiseptic wipe.
    • WHAT TO DO IN CASE OF BREAKAGE:

    • Turn off centrifuge immediately
    • Leave the lid closed for at least 30 minutes
    • Inform the College safety officer of breakage and complete Acident Report Form
    • Clean up and disinfect centrifuge with EXTREME CAUTION, wear approprite protective clothing
    • Place a warning notice on the centrifuge
    • NOTE: THE CENTRIFUGE BUCKET CONTAINING THE BROKEN SAMPLE SHOULD NOT BE OPENED UNTIL STERILIZED BY AUTOCLAVING
    • HANDLING BONE TISSUE:

      Bone tissue presents a particular hazzard as sawing such tissue can result in the release of aerosols, keeping the sample wetted will reduce the production aerosols, sawing under a wet cloth if appropriate will significantly reduce risk and the use of a face mask will further reduce risks. If sawing human bone tissue specific advice should be sort before starting work.

      CELL CULTURE:

      In laboratories were cell culture is undertaken it is essential that the highest standards of laboratory practice are followed

      GENERAL- Cell culture facility (Physics):

    • The area MUST be labelled as a biohazard
    • Only AUTHORIZED PERSONS are permitted to enter cell culture fascilities
    • Goood laboratory practice MUST be followed at all times
    • Cleanliness is esssential in cell culture, and following protocols for the maintaining this is required of all workers
    • Remove gloves and apply alcohol gel BEFORE leaving the cell culture room. Hands should be then be washed at sink area
    • A lab coat and Latex gloves MUST be worn
    • Keep work surfaces, incubator, equipment (pipettes, bottles etc) and microbiological safety cabnet sterilized using 70% alcohol
    • All human/animal tissue MUST be handled in a spill tray
    • All samples MUST be clearly labelled with sample details, name and date
    • Any spillages MUST be dealt with IMEDIATELY
    • All Waste MUST be disposed of in a safe manner:
    • WASTE DISPOSAL

    • Place syringes/needles etc in sharps bin. Pipettes can be placed in large sharps bin
    • All waste that has come in to contact with cell cultures or media MUST be disposed of in the following manner: (All waste cultures must be treated with Virkon (1%)and left for 20 minutes in a designated area to render it safe before disgarding) All waste must then be placed in the clinical waste bag for incineration. ALL BIOHAZARD WASTE MUST BE TREATED ON THE DAY OF USE)
    • MICROBIOLOGICAL SAFETY CABINETS

    • All cabinets should be well maintained and checks made by a professional engineer for correct operation and containment
    • Keep all surfaces and conents sterilized by spraying with ethanol (70%)
    • Keep working surfaces as clear as possible
    • DO NOT THE AIR INTAKE VENT to avoid contamination
    • Follow the manufacturer's instructions for the correct use of the cabinet
    • The correct class of cabinet MUST be used for the task in hand:

    • Class 1 safety cabinets these minimised the escape of aerosols by drawing air in through the open front. the air then passes through a high efficiency particulate filter (HEPA) and is then discharged (usually outside) NOTE: this class of cabinet protects the worker and NOT the work
    • Class 2 safety cabinets Air drawn through the front of cabinet is drawn downwards through grills at the base of the cabinet it then passes through a HEPA filter at the back of the cabinet into the cabinet circulating downwards through the working area providing a curtain of filtered air. These cabinets provide protection for the worker and the work from contamination and are particularly useful in cell culture.
    • Class 3 and Class1/3 safety cabinets these provide a totally enclosed working area and the operator wears long rubber gloves attached to the front panel.Incoming air is HEPA filtered once and outgoing air twice. Both the working and the work is protected from contamination. This type of cabinet is normally only necessary in containment level 3/4 laboratories.
    • Laminar flow cabinets These should be used for preparing media and pouring plates ONLY. The direction of air flow is outwards towards the worker, this provides a serile working environment BUT there is NO protection to the worker. THESE CABINETS SHOULD NEVER BE USED FOR HANDLING ANY PATHOLOGICAL MATERIALS OR CULTURES OF MICROORGANISMS
    • NITROGEN DEWARS: (for cell storage)

    • Store dewar in well ventilated area
    • Wear personal protective equipment (eye protection, insulated gloves, lab coat and appropriate footwear)
    • DO NOT put open ended pipes or tubes into liquid nitrogen
    • Handwashing Facilities

      Hand basins with hot and cold running water must be provided in all laboratory rooms. There should be at least one basin in each room. In rooms where a large number of people work, e.g. more than 10, it is desirable that two basins be provided. The basins should be near to the doors. Ideally, taps should be operable without needing to use hands.

      Automatic liquid soap dispensers should be avoided in favour of tablets of toilet soap regardless of the alleged economy of the former. Many of these dispensers must be operated by dirty or contaminated hands before they yield soap. It may not be obvious when they are empty.

      Paper towels in dispensers should be used. Roller towels are acceptable only if they are of the continuous flow type and are properly maintained and promptly replaced.

      As a general rule hands should be washed, preferably with a bactericidal soap, after completing any work and always before leaving the laboratory.

      Health

      Individuals with medical conditions which predispose them to infection (eg eczema, compromised immune systems, diabetes etc) are at a special risk. Everyone must notify their supervisor of any illness or other medical condition that may compromise the immune system and may make them more susceptible to hazards, which may arise through working with micro-organisms.

      Health surveillance is required under COSHH where:

      • there is an identifiable disease which may be related to workplace exposure;
      • there is a reasonable likelihood that exposure may happen;
      • there are valid techniques for detecting indications of the disease or its effects.

    Disinfection

    In General, disinfection is a less reliable means of sterilising materials than autoclaving (note: autoclaves are NOT available in physics). Disinfectants must be chosen carefully as there is no universal disinfectant, all have disadvantages. Disinfectants may deteriorate on standing or be inactivated by detergents, organic matter etc. and most are toxic or irritant. Some common types are listed below:

    Hypochlorite solutions:

    • Commonly recommended concentrations – 1,000 ppm for surface decontamination, 2,500 ppm for discard containers, 10,000 ppm for spillages.
    • These are active against bacteria (including spores) and viruses but have limited activity against fungi and tubercule bacilli.
    • Are compatible with anionic/non-ionic detergents but corrode many metals and damage rubber and are inactivated by organic materials and so need frequent changing.

    Chlorine releasing granules:

    • Usually contain sodium dichloroisocyanurate (NaDCC) and may also contain absorbent powders. They have a relatively long shelf life and are useful for spillages.

    Clear soluble phenolics:

    • Active against vegetative bacteria (including tubercule bacilli) but not active against spores and have a limited effect on fungi. They are not active against many viruses (particularly if not lipid containing).
    • Compatible with anionic/non-ionic detergents and metals and inactivated by rubber and some plastics
    • Hycolin has a new formulation and is not reliable.

    Glutaraldehyde:

    • Has similar range of activity to hypochlorites but should not corrode metal.
    • Does not readily penetrate organic matter and is relatively unstable once inactivated.
    • Is a potent allergic sensitizer.

    Alcohols:

    • Are normally used as 60-80% vv solutions in water.
    • Are active against protozoa and many viruses and vegetative bacteria (but not tubercule bacilli).
    • Can be used as a disinfectant skin rub (often with addition of 5% wv chlorhexidine)
    • Do not readily penetrate organic matter.
    • Are flammable.

    Virkon:

    • Claimed to be active against many types of organism.
    • Relatively non-toxic and non-corrosive.
    • This disinfectant is generally recommended

    Disposal of Biological Waste Material

    It is imperative that biological waste is handled and disposed of in a safe manner.Appropriate sterilisation procedures MUST be used prior to disposal, this is of particular importance where cell cultures have been used.

    Some guidelines:

    • All biological waste must be disposed of by incineration.
    • All body fluids and cell cultures should be treated with strong disinfectant and placed in sealed containers prior to disposal.
    • ALL BIOLOGICAL WASTE, TISSUES or FLUIDS, MUST BE DOUBLE BAGGED IN YELLOW WASTE BAGS AND SEALED WITH TAPE. THE BIOHAZARD BAGS MUST BE PLACED INTHE DESIGNATED YELLOW BINS (PHYSICS CAR PARK)

    Action in the event of spillage

    GENERAL INFORMATION

    When dealing with any spillage protective cothing including lab coat, gloves and apron should be worn.

    THE AREA MUST BE CLEARLY LABELLED AS A BIOHAZARD.

    There MUST be contingency plans for dealing with foreseeable emergencies. These could include spillage control, room evacuation, fumigation and decontamination and, if there is a risk of infection - first-aid and medical treatment (prophylazis) and health surveillance and counselling of exposed people.

    • Spillages should be contained and covered with disinfectant granules or absorbent paper/cloth soaked in disinfectant.
    • The disinfectant should be allowed to act for at least 15 minutes.
    • The debris should be swept gently into a dustpan using a piece of board or stiff card.
    • Any residual pieces of glass etc should be picked up with forceps or swabs.
    • Debris should be put in a suitable container for disposal by a safe route; and
    • Further disinfectant should be applied to contaminated surfaces.
    • Rooms must not be re-occupied until it is safe to do so.

    SPILLAGE OF BLOOD OR OTHER BODY FLUIDS:

    All items required for dealing with spillages etc must be readily available and all workers must know the procedures.

    A BLOOD SPILL KIT should always be readily availible when ever blood is being used.

    This consists of the following:

  • Chlorine tablets (e,g. Klorosept 87 or Actichlor) to be used to make up a 10,000 ppm active chlorine solution.
  • Measuring cylinder or jug
  • Antiseptic wipes
  • Disposable latex gloves
  • Disposable apron
  • Small biohazard waste bag
  • BLOOD CENTRIFUGATION

    Centrifugation of blood MUST only be undertaken by trained individuals following the laboratory protoco. Only sealable centrifuge buckets with tight fitting lids may be used for blood centrifugation. A Blood spill kit must be available and personal protective equipment worn.

    ACCIDENTS

    In the event of an accident/incident/emergency, immediate steps must be taken to mitigate the consequences. Only people essential for carrying out repairs and other essential work may be permitted in the affected area and they must be provided with appropriate personal protective equipment.

    If there is a risk of airborne infection the laboratory must be evacuated as quickly as possible. It may be necessary to fumigate the room before reoccupation.

    People attending casualties should avoid becoming contaminated themselves.

    • Contaminated clothing must be removed as quickly as possible.
    • Remove contamination of skin/eyes/mouth by thorough washing with clean water.
    • Eyes should not be rubbed nor skin scrubbed.
    • Small puncture wounds should be encouraged to bleed; minor cuts and similar lesions should be washed with soap and water or a suitable detergent before being thoroughly washed and dressed; and
    • Medical advice must be sought if there is a risk of infection.

    HOW TOP AVOID NEEDLE STICK INJURIES:

    Risk

  • This type of injury is of particular concern as it poses a risk of transmitting infection, particularly where human blood or other tissues are concerned
  • The major blood borne viruses of concern are hepatitis B and C and human immunodeficiency virus (HIV)
  • Preventative measures

  • Cover any exsisting wound with waterproof dessing
  • ALWAYS wear personal protective clothing (gloves/apron) and sturdy footwear when handling sharps
  • Ensure that you are vacinated against Hepatitis B
  • NEVER re-sheath needles!
  • ALWAYS handle and carry sharps in a tray or other suiable container
  • Clean up blood spills immediately and disinfect area
  • ALWAYS wash your hands after dealing with Sharps
  • In case of injusy: Encourage bleeding, wash skin throughly, apply waterproof dressing, SEEK MEDICAL ATTENTION AND REPORT ACCIDENT IMMEDIATELY If injusy has occured and there is risk of infection, health surveillance may be necessary.

    Containment measures

    JOHN CAN YOU PUT THIS SECTION AS A SUBDIRECTORY

    The term “containment” describes the way in which biological agents are managed in the laboratory environment so as to prevent or control the exposure of laboratory workers, other persons and the outside environment to the agent(s) in question. This is achieved by a combination of measures.

    Primary containment, ie, the protection of the worker and the immediate environment can be achieved through a combination of good microbiological practices/ techniques and the use of appropriate safety equipment, eg, microbiological safety cabinets. Further protection may be achieved through the use of appropriate immunisations, although this should be seen only as a useful supplement to reinforce procedural controls and the use of safety equipment, not the sole protective measure.

    Secondary containment, ie, the protection of the people and the environment outside the laboratory can be achieved by a combination of laboratory design, engineering controls and operating procedures.

    Containment measures must be reviewed at suitable, regular intervals and immediately if there is reason to suspect that the measures are no longer adequate or, if in the light of new scientific or technical knowledge, the assessment is inadequate. Laboratory containment measures must reflect the nature and severity of the biological hazard; an outline guide is provided in the table below.

    Laboratory Containment Level

    Containment Measures

    1

    2

    3

    Laboratory suite: isolation

    Not required

    Not required

    Required

    Laboratory sealable for fumigation

    Not required

    Normally required, depending on the workplace risk assessment

    Required

    Equipment

    Surfaces impervious to water and resistant to acids, alkalis, solvents, disinfectants, decontamination agents and easy to clean

    Required for bench

    Required for bench and floor

    Required for bench, floor, ceiling and walls

    Entry to lab via airlock

    Not required

    Not required

    Required

    An inward airflow into the laboratory (negative pressure) must be maintained by extracting room air to atmosphere

    Not required

    Required where indicated in risk assessment

    Required

    Extract and input air from the laboratory must be HEPA filtered (H14 standard tested to 99.997% efficiency)

    Not required

    Not required

    Single HEPA filters required for extract air, single HEPA filters for input air

    Use of a microbiological safety cabinet/enclosure

    Not required

    Required where there is a risk of aerosol generation

    Required, Class I/III cabinet or isolator

    Autoclave

    Required on site

    Required in the building

    Autoclave required in the laboratory

    System of Work

    Access restricted to authorised personnel

    Required

    Required

    Required (key pad lock required)

    Specific measures to control aerosol dissemination

    Not required

    Required so as to minimise

    Required so as to prevent escape from primary containment

    Shower

    Not required (unless required for chemical safety)

    Not required (unless required for chemical safety or if large volumes/high concentrations to be used)

    Required – emergency use

    Protective clothing

    Suitable protective clothing required

    Suitable protective clothing required

    Suitable protective clothing required; footwear required where and to the extent the risk assessment shows it is required

    Use of disposable gloves

    Required where indicated in risk assessment

    Required where indicated in risk assessment

    Required

    Efficient control of disease vectors

    Required where and to the extent the risk assessment shows it is required

    Required

    Required

    Specified disinfection procedures in place

    Required

    Required

    Required

    Biohazard sign displayed on laboratory door, fridges, freezers and transport containers

    Required

    Required

    Required

    Waste

    Inactivation of biohazards in effluent from hand-washing sinks and showers and similar effluents

    Not required

    Not required

    Required where and to the extent the risk assessment shows it is required

    Inactivation of biohazards in contaminated material and waste

    Required by validated means, where and to the extent the risk assessment shows it is required

    Required by validated means

    Required by validated means

    Other Measures

    Laboratory to contain its own equipment

    Not required

    Not required

    Required, so far as is reasonably practicable

    An observation window or alternative is to be present so that occupants can be seen

    Required where and to the extent

    the risk assessment shows it is required

    Required where and to the extent the risk assessment shows it is required

    Required

    Safe storage of biohazardous material

    Required where and to the extent the risk assessment shows it is required

    Required

    Secure storage required

    Telephone or intercommunication system between the laboratory and the clean outside area

    Required

    Required

    Required

    Written records of staff training

    Required

    Required

    Required

    Space for laboratory workers

    There must be adequate space (at least 11m³) in the laboratory for each worker

    There must be adequate space (at least 11m³) in the laboratory for each worker

    There must be adequate space (at least 24m³) in the laboratory for each worker

    Reference material

    JOHN CAN YOU PUT THIS AS A SUB DIRECTORY

    • Health and Safety at Work Act (HASWA) 1974 and the European Communities Act 1972 and regulations within, including Control of Substances Hazardous to Health (COSHH) 2002 Regulations (as amended)
    • The Management, Design and Operation of Microbiological Containment Laboratories. Advisory Committee on Dangerous Pathogens. HSE Books 2001
    • Biological agents: managing the risks in laboratories and healthcare premises. Advisory Committee on Dangerous Pathogens. HSE Books 2005.

    http://www.hse.gov.uk/biosafety/biologagents.pdf

    • Road and Rail Transport – ADR European Agreement concerning the International Carriage of Dangerous Goods by Road

    RID Regulations concerning the International Carriage of Dangerous Goods by Rail

    The Carriage of Dangerous Goods and Use of Transportable Pressure Equipment Regulations 2004 (SI 568) as amended

    • Maritime Transport - IMDG International Maritime Dangerous Goods (Code)

    The Merchant Shipping (Dangerous Goods and Marine Pollutant) Regulations 1997 (SI 2367)

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