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Pathogens and Land
             Contamination
               YCLF, 17th February 2012
Dr Russell Thomas, Parsons Brinckerhoff Ltd, Bristol, UK
                          and
Dr Brian Crook, Health and Safety Laboratory, Buxton, UK




                                                           1
Pathogens and Land Contamination
                                   Microorganisms and Pathogens
• Microorganisms – are distributed throughout soil, air and water.
• Most don’t pose a risk to healthy humans – many millions inhabit our bodies.
• Microorganisms able to inflict “damage” on host organisms are called pathogens.
• Likelihood of infection is dependent on many factors including:
  1.    Standards of sanitation and hygiene;
  2.    Quality of diet and drinking water;
  3.    Health of the individual
       • Do they have pre-existing conditions/diseases (e.g. cystic fibrosis, diabetes);
       • Do they have a weakened immune system (very young or old, immuno-compromised);
       • Do they have cuts/grazes/burns to skin;
  4.    Prevalence of local diseases and sources of infection; and
  5.    Immunisation status.
• Factors 1,2,4 and 5 are generally not major issues in the UK.
• Most of the worst pathogens are not endemic or have been eradicated in the UK.


                                                                                           2
Pathogens and Land Contamination
                                             Likelihood of Infection
• Most people’s interactions with pathogens are limited to common illnesses e.g.
 • Colds and flu;
 • Sickness and diarrhoea; and
 • Food poisoning.
• These are generally caused by viruses (e.g. influenza virus) and bacteria.
• Fungi and protozoans can also be pathogenic.
• Pathogens are mainly an issue for public health, but specific illnesses may relate
  to land and building.



                            +
• Exposure to certain pathogens may increase due to a person’s living or working
  environment.
• For example, living or working on land potentially contaminated by pathogens.

                                                                                   3
Pathogens and Land Contamination
                                                       Potential Pathogens

 • Most pathogens are unlikely to be encountered in soils as they do not remain
   viable outside of a host organism for a significant length of time.
 • Exceptions are certain types of spore-forming bacteria.
 • Certain bacteria form spores, a resting (dormant) state in which they survive until
   conditions are suitable for them to return to their normal living (vegetative) state.


Source: Public Health Image Library (PHIL)
                                             • The primary spore forming bacteria of
                                               concern are:

                                              • Spore forming Clostridium Sp. e.g.
                                                Clostridium tetani and Clostridium
                                                perfringens
                                              • Bacillus anthracis, the causative agent of
                                                anthrax
                                               Left: An electron micrograph of anthrax spores

                                                                                                4
Pathogens and Land Contamination
                                                 Other Potential Pathogens
• Other examples of pathogens which may be                               Source: Public Health Image Library (PHIL)
  encountered are:
• Bacteria (non spore forming bacteria)
    • Legionella pneumophila – cause of Legionnaires disease.
    • Leptospira – cause of Weil’s Disease.
    • Pseudomonas sp. – causes minor infections in healthy humans,
      but is more serious, possibly fatal in the immuno-compromised.
•   Fungi
    • Aspergillus sp. - causes farmers lung and other diseases, mainly
      allergic responses to spores and conditions can become chronic.    Source: Public Health Image Library (PHIL)
      Particularly harmful to those who are immuno-compromised.
•   Protozoans
    • Cryptosporidium parvum – causes cryptosporidiosis, the main
      symptom of which is very bad diarrhoea.
•   Viruses
    • Variola major virus – caused the disease Smallpox.
• Prions
    • BSE/variant Creutzfeldt Jacob Disease.

                                                                                                                      5
Pathogens and Land Contamination
                      Sites Potentially at Risk from Pathogens
Sites potentially affected                              Processes undertaken/ Examples of potential pathogens
Slaughterhouses, abattoirs, knackers yards    Animal slaughter and processing into various products incl. meat, hides etc. e.g.
and fellmongers.                              Spore forming bacteria, BSE/vCJD.
Rendering plants, glue works and bone         Bones and inedible offal used to produce glue, gelatine, oils and fertilisers. e.g.
processing sites.                             Spore forming bacteria incl. B.anthracis.
Tanneries and related hide processing sites   Hides prepared for use in leather goods, e.g. jackets, gloves and shoes. e.g.
                                              Spore forming bacteria, incl. B.anthracis.
Meat processing and packaging factories       Processing and packaging of butchered meat. e.g. Spore forming bacteria, BSE.
Human burial sites, crypts, catacombs,        Burial of infected bodies, particularly semi preserved infected bodies in lead
graveyards etc.                               coffins and coffin liquors. e.g. Smallpox, vCJD, C. Tetani, Leptospirosis.
Animal burial sites                           Mass graves of buried animals from major outbreaks of disease. e.g. Spore
                                              forming bacteria, BSE, C. Tetani, Leptospirosis.
Woollen mills                                 Wool/hides contaminated by B.anthracis spores.
Old buildings                                 B. anthracis spores in horse hair plaster/building fabric
Water pipes in contaminated ground            Microorganisms can grow (in water pipes/tanks) on organic compounds diffusing
                                              into non chemically resistant pipes. e.g. Pseudomonas sp.
Air conditioning systems and stagnant water   Stagnant water in pipes and air conditioning systems at risk from Legionella.
in pipes/tanks
Agricultural land and water courses           Risks from pathogens in food storage (Aspergillus sp.), slurry, bovine urine, rats
                                              and water course (Cryptosporidium parvum and Leptospirosis).


                                                                                                                                6
Pathogens and Land Contamination
      Examples of Pathogens – Bacillus anthracis
• Bacillus anthracis causes the potentially fatal disease anthrax.
• It is a zoonotic pathogen and predominantly infects herbivorous animals.
• Humans can be infected, but are relatively resistant.
  Lungs of patient after 4 months of anthrax
  infection in the lung. Source: Public Health
  Image Library (PHIL)




                                                 Dermal Anthrax lesion, source: Public Health Image Library (PHIL)


                                                                                                                     7
Pathogens and Land Contamination
               Anthrax Infections how do they Occur



                                 2. Germination and
                                 multiplication of anthrax in
                                 lymphatic system and spleen.
                                 Vegetative forms released in
                                 large numbers into blood in
                                 final hours of life.
3. Vegetative Spores, released
at death as haemorrhagic                                        1. Cow ingests soil
exudates from nose, mouth or
anus in spilt blood




                                  Spores - Sporulate on
                                  exposure to oxygen

                                                                Anthrax spores from previous
                                                                anthrax deaths or buried
 infection of further soil                                      infected carcass.

                                                                                               8
Pathogens and Land Contamination
                      Anthrax Infections how do they Occur
             1. Pulmonary anthrax, inhalation of spore laden dust
             from the site, entering the lungs.                          Source: Public Health Image Library (PHIL)




                                                                     2. Cutaneous infection via a lesion
3. Gastrointestinal, only caused by eating                           or cut in the skin.
infected meat or contaminated water. Has
not occurred in the UK in past 100 years




                 Anthrax spores from previous
                 anthrax deaths or buried
                 infected carcass.

                                                                                                                      9
Pathogens and Land Contamination
         Prevalence of Bacillus anthracis in the UK
                             Anthrax outbreaks in UK livestock: 1887 to 2006            (Source DEFRA)
               1600
               1400
               1200                                                                     Number of outbreaks
               1000
                800
                600
                400
                200
                 0
                      1887


                                1897


                                       1907


                                              1917


                                                     1927


                                                            1937


                                                                   1947


                                                                          1957


                                                                                 1967


                                                                                           1977


                                                                                                  1987


                                                                                                         1997
• Anthrax was more prevalent in humans in the UK due to a greater incidence
  in livestock and also greater workplace exposure (e.g. woollen mills).
• The number of human cases has dropped steadily over the years, with very
  few cases since the 1970’s, workers in key industries vaccinated since 1965.

                                                                                                                10
Pathogens and Land Contamination
               General routes of infection for Anthrax

Most new cases                   Gardens
of anthrax from
    imported                                            Humans
                                Animal feeds
   materials
               Bonemeal                           Animals
     Imports                   Leather
                Hides,         work,
                Wool, Hair     tanneries,            Land or water
                               wool
                                               Process
                               processing.
                                               wastes
                               etc.            Industrial
   Cases can occur                             Anthrax
     from historic
        anthrax              Historic
    contamination            contamination

                                                                       11
Pathogens and Land Contamination
                                                 Anthrax Infections
• Today anthrax occurs sporadically with isolated cases reported in humans.
• Cases have usually been dermal anthrax. Black boil-like lesions develop on the
  skin, readily diagnosed and easily treated.
• Rarely inhalation anthrax occurs, resulting in haemorrhagic pleural effusions,
  severe septicaemia, meningitis and high death rates. Recent case caused by
  imported hides contaminated with anthrax spores, used make into drums.
• In 2009-2010 a number of cases of death from anthrax occurred to the
  intravenous injection of Heroin cut with anthrax.




                                                                              12
Pathogens and Land Contamination
                                   Viral Infection - Variola Major Virus
• Smallpox is caused by Variola major virus, which has since been eliminated
  from the world.
• There is no treatment and it has high mortality rates.
• The last outbreaks were mainly in the 1930s, some sporadic cases until the
  1970’s but none afterwards.
• Smallpox can survive for long periods of time in dry scabs (13 years has
  been documented).
• In normal conditions, it is unlikely to survive for more than 48 hours.
                                            • Intact virus was recovered from an exhumed body
                                              (Spitalfields 1985), over 100 years old.
                                            • The virus could not be grown in the laboratory and
                                              not considered infective.
                                            • The risk that smallpox might re-emerge if the remains
                                              of smallpox victims are disturbed is remote, but
Smallpox infection. Source: Public Health
Image Library (PHIL)                          clearly the impact would be very significant.

                                                                                                13
Pathogens and Land Contamination
                                   Leptospirosis – Weils Disease
• Leptospirosis, a bacterial disease passed from animals to humans (zoonoses).
                                                              Leptospirosis cell within human kidney.
• It can be caught from:                                      Source: Public Health Image Library (PHIL)


 • work on farms or handling animals (e.g. cows and rats);
 • contact with canals, rivers and other watercourses; and
 • work in drainage ditches and sewers;



• Infection via contact with infected animal (particularly rats) urine, fluids, tissue or
  indirect exposure through water, soil or animal foodstuffs.
• The bacteria enter the body through cuts in the skin, the skin lining the mouth and
  nose, and membrane at the front of the eye.
• Treated early the prognosis is good, if untreated it can cause liver and kidney failure
  and be fatal.


                                                                                                           14
Pathogens and Land Contamination
           Assessing the Risk – What is Acceptable?
• Pathogens are present throughout the environment, but there are different risks
  posed by different pathogens.
• Humans assess the risks posed by pathogens everyday:
  • Should we go into work when our colleagues have a cold?
  • Gauging the cleanliness of eating establishments?
  • Wash your hands after using the toilet?
  • What is the water quality at a beach?                         Source: Public Health Image Library (PHIL)


• Your bodies immune system is constantly battling against potential pathogens.
• When dealing with soils/buildings or water, the risks have to be taken in the context
  of the threat posed by the likely pathogens and their abundance.
 • e.g. Anthrax – Highly dangerous and major economic threat to agriculture. No risk is
   acceptable as only one organism may be required to cause infection; no detectable viable
   spores/cells is threshold.
 • e.g. Leptospirosis/Clostridium – potentially abundant in environment, minimise exposure to
   sensitive receptors as it is impossible to eradicate pathogen.


                                                                                                        15
Pathogens and Land Contamination
                          The Unique Problem of Pathogens
                                                                Carcinogenic
                                                                effect on cells




  Acutely toxic                      Pathogenic                          Rapid
  substance,                         Bacterial                           division of
  the response                       cell division                       cancer cells
  relates to dose




One cell divides into two, two cells into four, four cells into eight, this process continues leading
to the growth millions of cells. This is analogous to cancer and the effect of carcinogens.

                                                                                                   16
Pathogens and Land Contamination
                                            Viral Infection and Replication
                 2. Virus penetrates cell
                 membrane an injects
                 nucleic acid (DNA and
                 RNA) into cell.


                    Coated vesicle
                                                   1. Viruses attach to cell
Endosome



                   mRNA Translation


                                                                                  nucleic acid (DNA and RNA)
  Nucleus                            Synthesis of
                                     envelope proteins


     Endoplasmic
     reticulum

                          Golgi apparatus
                                                           4. New viral nucleic acids are
            3. Viral nucleic acid                          packaged into viral particles and
            replicates using host                          released from the host cell, which
            cellular machinery.                            may be destroyed in the process.




                                                                                                               17
Pathogens and Land Contamination
   Risk Assessment – Sources of Contamination
• Industrial Sites
• The areas of sites that processed animals or products made from animals, posing
  the greatest risk of pathogen contamination are:
  • Areas where carcasses or hides/wool were received and stored;
  • Areas for the washing or early processing of the materials and associated drainage
    systems; or
  • Areas used for waste disposal, animal burial or where land raising has taken place.
  • Areas inhabited by pests (e.g. rodents) which are vectors of disease (applies below)
• Targeting these areas would be the most effective approach to detecting pathogens,
  if free of pathogens it is likely that others are also uninfected.
• Agricultural Sites
• Does information suggests previous disease outbreaks or animal burials on site,
  “Animal Health and Veterinary Laboratories Agency” should be contacted.
• If burial sites (due to notifiable diseases) are recorded it is unlikely that
  investigation/development of these areas would be allowed.
• Has blood, tissues or anaerobic digester waste been spread on the site.

                                                                                           18
Managing Risks from Gas Industry Construction in Contaminated Land
                                   What/Who Might be at Risk
                     Surface Water Drainage Inhalation, ingestion or dermal contact of Surface
Public/neighbours animalslivestock         soils/dusts, containing microorganisms or Water
The very young                              spores.                                    Courses
The elderly
The ill
                                                                            Building
                                      Construction                         occupants
                                         workers           The Old Tannery




             Buried Animals


Groundwater      Leaching of Microorganism and spores into, drains, surface
                 water and groundwater.

                                                                                          19
Pathogens and Land Contamination
             Risks from Investigation and Construction
• The risks from pathogens may be overlooked through ignorance or because they are
deemed outside the scope of a project (e.g. Part 2a).
• Risks may be higher on sites which are not potentially polluted by chemical
pollutants and protection not offered by PPE.
• Health and Safety issues which may be overlooked include:
 • Not including the analysis of pathogens;
 • Not identifying the risk of potential pathogens;
 • Adopting suitable drilling methods that minimise waste, aerosols,
 and dust generation;
 • Provision of suitable PPE including, the covering of cuts and
 respiratory protection;
 • The safe handling, storage and disposal of waste materials;
 • Quarantining samples until cleared of expected pathogens; and
 • Good site hygiene practices.



                                                                                     20
Pathogens and Land Contamination
       Assessing the Risk – Other Considerations
• The three main routes of infection from pathogens are: dermal contact, inhalation
  and ingestion.
• However, the main route of infection varies between different pathogens; the
  severity of the disease may also change dependant on the route of infection.
• Zoonotic pathogens cause a greater risk to their target host animals, although
  they can still cause severe disease in humans.
• Children are at the greatest risk of infection and serious illness from
  environmentally transmitted pathogens because:
 • Absent or poor sanitation habits and more likely to ingest soil;
 • Immature immune system; and
 • Intestinal mucosa more permeable to water and reduced stomach acid and pepsin
   secretion
• The elderly and immuno-compromised are also at greater risk from pathogens.
• Pathogens may also migrate from the soil into the drains or underlying aquifer;
  quantifying these risks is also problematic.
                                                                                    21
Pathogens and Land Contamination
        Assessing the risk – Scottish Borders Case
• This zero risk approach was taken following a confirmed case of inhalation anthrax
  in the Scottish Borders.
  • The source of the anthrax had been traced to a drum made from an imported infected hide.
  • Some sites where this drum had been used were later found to be contaminated with
    anthrax spores.
  • Affected sites were decontaminated until no viable spores were detected.
  • This ensured there was no risk public health for these sites, albeit at a cost of £460k (£227k
    - decontamination contract).
• Estimates of the human LD50 for inhalation anthrax range from 4,100-10,000
  spores, other studies have shown humans can be exposed to around 1,000 spores
  without contracting anthrax.
• Despite the growing evidence, there are still gaps in the dose response relationships
  between anthrax exposure and infection, especially for dermal infections.
• If this information is provided then a risk based approach may be possible.
• But one cell is theoretically all that is required for infection.

                                                                                               22
Pathogens and Land Contamination
                                           Pathogens and Legislation
• Planning (PPS 23: Planning and Pollution Control, England)
 • PPS23 describes the hazards posed by a contaminant to include biological (pathogens) and
   therefore sites at risk from pathogens proceeding through the planning system requires an
   investigation to consider the risks posed by pathogens during and after development.

• Environmental Liabilities Directive and Environmental Damage Regulations
• The definition of environmental damage in respect of 'land damage' is "any land contamination
  that creates a significant risk of human health being adversely affected as a result of the direct
  or indirect introduction, in, on or under land, of substances, preparations, organisms or micro-
  organisms".
• The Control of Substances Hazardous to Health (2002) Regulations
 • COSHH requires that any risk associated with biological agents must be considered prior to
   worker activity that might place employees or the public at risk; this includes the investigation
   and remediation of potentially contaminated land.
• Pathogens included in EU/UK waste legislation, Hazard H9 - “Infectious”
• Pathogens can be dealt with through Statutory Nuisance legislation.


                                                                                                  23
Pathogens and Land Contamination
                                           Pathogens and Legislation
• Public Health (Control of Disease) Act 1984, (substantially amended by the Health
  and Social Care Act 2008) and various Health Protection Regulations 2010, including
  the Health Protection (Part 2A Orders) Regulations 2010:
 • Local authorities and JP to take action to protect public health from a risk of significant harm
   from infection or contamination. Registered medical practitioners and laboratories, to notify a
   specific list of notifiable infectious diseases and causative agents and also other infections or
   contamination, such as with chemicals or radiation, that may pose a significant risk to public
   health.
• Specified Animal Pathogens Order 2008 (SAPO), A person must immediately notify
  a veterinary inspector if:
 • has possession of any thing in respect of which they have reasonable grounds for suspecting
   that an animal pathogen specified in Part 1 of Schedule 1 (e.g. B.anthracis) is present and
   does not have a licence in respect of that pathogen.
• Notifiable diseases in animals must be reported to the Animal Health and Veterinary
  Laboratories Agency.
• Notifiable diseases in humans must be reported to the local Consultant in
  Communicable Disease Control and under RIDDOR.
• Pathogen specific legislation, e.g. The Anthrax Order 1991
                                                                                                 24
Pathogens and Land Contamination
                                                  Part 2A and Pathogens
• Pathogens are not included or excluded from the definition of a “substance”.
• A substance is defined in section 78A(9) of Part 2A of the Environmental Protection
  Act 1990 as: “any natural or artificial substance, whether in solid or liquid form or in
  the form of a gas or vapour”.
• Can a pathogen be classed as a ‘substance’ under Part 2A?
- the definition of a substance is unspecific and unclear; and
- the definition of a substance in Part 2A has not been looked at in case law.

• The position of the Environment Agency was that microorganisms are not mentioned
  in the definition of a “substance” in Part 2A, whereas in other legislation they are.
• They believe that their omission from Part 2A was intended and that pathogens
  should not be considered a “substance” under Part 2A, and therefore should fall
  outside the remit of Part 2A.
• However – the decision lies with the local authority, only they are able to determine a
  site to be contaminated land based on the presence of pathogens under Part 2A!

                                                                                         25
Pathogens and Land Contamination
                                             Analysis of Pathogens

• Tests for many pathogens have been developed for detection from human or
  animal bodily fluids; they are not optimised for detection from soils/water.
• Some tests used for detection in soil/water samples do not accurately quantify
  pathogen numbers (cells/spores/viruses).
• With the exception of Legionella, there are a limited number of laboratories that
  can test for pathogens from environmental samples.
• Seek advice from experienced microbiologists.
• Tests must be carried out in specialist microbiology facilities;        B. anthracis
  requires Level 3 Containment.
• Analytical methods are based one or both of:
 • culturing, growing cells in broth or agar plates, or growing virus in tissue culture
 • molecular detection, analysis of RNA or DNA.


                                                                                      26
Pathogens and Land Contamination
                                              Analysis of Pathogens
• For the rapid selective isolation and detection of Bacillus anthracis in potentially
  contaminated materials (e.g. animal hair or soil) the HSL uses a combination of
  enrichment culture and molecular detection by polymerase chain reaction
  (PCR).
• Culture takes overnight and PCR, which tests for unique DNA sequences from
  the B. anthracis toxin genes, takes 4-6 hours.
• Similar approaches can be used for the detection of spore forming Clostridium
  Sp. and opportunistic pathogens such as Pseudomonas Sp. in soils, water and
  building material.
• Legionella testing in water is commonly undertaken by commercial
  environmental Labs; this is a culture based test.
• Leptospira can be analysed in water samples although it is a lengthy process
  (6 weeks) and not recommended for large surface water bodies. It is feasible to
  undertake PCR based tests. Testing in humans is done by the HPA in Hereford.
• The detection of Smallpox in humans could only be undertaken at a specialist
  facility such as (HPA Porton Down)

                                                                                         27
Pathogens and Land Contamination
                     Factors to Consider for PCR Testing

• Molecular detection by PCR is
rapid (hours), sensitive (our
anthrax PCR detects down to less
than 10 spores) and specific…..

BUT:                                               Source: Public Health Image Library (PHIL)



• Molecular tests can be affected by interferents such as humic acids and organic
debris – important for environmental/soil samples to have a validated clean-up step.
• A positive result shows that DNA sequences are present, not necessarily live micro-
organisms. Which is why we use culture as a back-up. However a rapid PCR positive
result can be used as an alert for countermeasures or to focus further tests.
• The assay results are only as good as the sample taken.

                                                                                                28
Pathogens and Land Contamination
                                                    Sampling Strategies
• Statistically based strategies:
• For example, Visual Sample Plan (VSP) – Pacific Northwest National Laboratory
  and US Dept of Energy http://vsp.pnnl.gov/ - a software tool for optimal sampling
  design and statistical analysis for environmental monitoring, characterisation and
  remediation, response to CBRN events, sampling of soils, buildings, groundwater,
  sediment, surface water, subsurface layers.
• Favored by Government Decontamination Service for sampling to support clean up.
• Targeted/intelligence led sampling:
• Be prepared to step outside the Plan-led sample collection – in the Scottish anthrax
  case the main focus of contamination was one floor mat. Use your eyes and
  imagination.
• Supplement statistical sample plan with site specific information.
• Details of past history and use of site. Talk to ex employees or residents.
• Previous location of buildings, activities, effluent drains

                                                                                       29
Pathogens and Land Contamination
                                 Bioaerosol Sampling

• Examples of fixed point and personal sampling methods for
  measuring bioaerosol emissions:
• Andersen impactors;                    Liquid impingers;
•




                                                              30
Pathogens and Land Contamination
                                    Bioaerosol Sampling

• Filtrations samplers (fixed point and personal):




                                                                31
Pathogens and Land Contamination
                                                   Worker Protection
• Levels of live pathogens are likely to be low.
• General personal protection for outdoor work, including gloves, covering
  cuts and grazes and where required respiratory protection, likely to provide
  sufficient protection.
• During building remediation or demolition, asbestos protection also
  sufficient protection against pathogens (with safe disposal of contaminated
  PPE).
• This should be supplemented by a procedure for recording exposures,
  injuries and any subsequent ill-health.
• Suitable secure containers for storage of samples and transportation to
  analyst.
• Adequate information accompanying samples to ensure safe handling for
  analysts.
                                                                            32
Pathogens and Land Contamination
                                       Future Considerations
• Continued emergence of non-fossil fuel energy technologies.
• Examples include mechanical & biological treatment (MBT), waste or
  biomass to energy, anaerobic digestion (AD).
• Potential for biological and chemical residues from the process
  contaminating surrounding soil, leachate run-off, bioaerosol emissions,
  also asphyxiant gases in confined spaces.
• Also subsequent spreading of wastes, such as AD waste, to land needs
  to be managed carefully.
• Scaled-down waste to energy projects (e.g., community heat and power)
  may be less well managed.




                                                                            33
Pathogens and Land Contamination
                  Thanks for your Attention
  Dr Russell Thomas, Technical Director
        Parsons Brinckerhoff Ltd
 thomasru@pbworld.com Tel: 0117 9339262

Dr Brian Crook, Microbiology Team Leader
       Health and Safety Laboratory
 brian.crook@hsl.gov.uk Tel: 01298 218421

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Pathogens And Land Contamination Yclf Unan

  • 1. Pathogens and Land Contamination YCLF, 17th February 2012 Dr Russell Thomas, Parsons Brinckerhoff Ltd, Bristol, UK and Dr Brian Crook, Health and Safety Laboratory, Buxton, UK 1
  • 2. Pathogens and Land Contamination Microorganisms and Pathogens • Microorganisms – are distributed throughout soil, air and water. • Most don’t pose a risk to healthy humans – many millions inhabit our bodies. • Microorganisms able to inflict “damage” on host organisms are called pathogens. • Likelihood of infection is dependent on many factors including: 1. Standards of sanitation and hygiene; 2. Quality of diet and drinking water; 3. Health of the individual • Do they have pre-existing conditions/diseases (e.g. cystic fibrosis, diabetes); • Do they have a weakened immune system (very young or old, immuno-compromised); • Do they have cuts/grazes/burns to skin; 4. Prevalence of local diseases and sources of infection; and 5. Immunisation status. • Factors 1,2,4 and 5 are generally not major issues in the UK. • Most of the worst pathogens are not endemic or have been eradicated in the UK. 2
  • 3. Pathogens and Land Contamination Likelihood of Infection • Most people’s interactions with pathogens are limited to common illnesses e.g. • Colds and flu; • Sickness and diarrhoea; and • Food poisoning. • These are generally caused by viruses (e.g. influenza virus) and bacteria. • Fungi and protozoans can also be pathogenic. • Pathogens are mainly an issue for public health, but specific illnesses may relate to land and building. + • Exposure to certain pathogens may increase due to a person’s living or working environment. • For example, living or working on land potentially contaminated by pathogens. 3
  • 4. Pathogens and Land Contamination Potential Pathogens • Most pathogens are unlikely to be encountered in soils as they do not remain viable outside of a host organism for a significant length of time. • Exceptions are certain types of spore-forming bacteria. • Certain bacteria form spores, a resting (dormant) state in which they survive until conditions are suitable for them to return to their normal living (vegetative) state. Source: Public Health Image Library (PHIL) • The primary spore forming bacteria of concern are: • Spore forming Clostridium Sp. e.g. Clostridium tetani and Clostridium perfringens • Bacillus anthracis, the causative agent of anthrax Left: An electron micrograph of anthrax spores 4
  • 5. Pathogens and Land Contamination Other Potential Pathogens • Other examples of pathogens which may be Source: Public Health Image Library (PHIL) encountered are: • Bacteria (non spore forming bacteria) • Legionella pneumophila – cause of Legionnaires disease. • Leptospira – cause of Weil’s Disease. • Pseudomonas sp. – causes minor infections in healthy humans, but is more serious, possibly fatal in the immuno-compromised. • Fungi • Aspergillus sp. - causes farmers lung and other diseases, mainly allergic responses to spores and conditions can become chronic. Source: Public Health Image Library (PHIL) Particularly harmful to those who are immuno-compromised. • Protozoans • Cryptosporidium parvum – causes cryptosporidiosis, the main symptom of which is very bad diarrhoea. • Viruses • Variola major virus – caused the disease Smallpox. • Prions • BSE/variant Creutzfeldt Jacob Disease. 5
  • 6. Pathogens and Land Contamination Sites Potentially at Risk from Pathogens Sites potentially affected Processes undertaken/ Examples of potential pathogens Slaughterhouses, abattoirs, knackers yards Animal slaughter and processing into various products incl. meat, hides etc. e.g. and fellmongers. Spore forming bacteria, BSE/vCJD. Rendering plants, glue works and bone Bones and inedible offal used to produce glue, gelatine, oils and fertilisers. e.g. processing sites. Spore forming bacteria incl. B.anthracis. Tanneries and related hide processing sites Hides prepared for use in leather goods, e.g. jackets, gloves and shoes. e.g. Spore forming bacteria, incl. B.anthracis. Meat processing and packaging factories Processing and packaging of butchered meat. e.g. Spore forming bacteria, BSE. Human burial sites, crypts, catacombs, Burial of infected bodies, particularly semi preserved infected bodies in lead graveyards etc. coffins and coffin liquors. e.g. Smallpox, vCJD, C. Tetani, Leptospirosis. Animal burial sites Mass graves of buried animals from major outbreaks of disease. e.g. Spore forming bacteria, BSE, C. Tetani, Leptospirosis. Woollen mills Wool/hides contaminated by B.anthracis spores. Old buildings B. anthracis spores in horse hair plaster/building fabric Water pipes in contaminated ground Microorganisms can grow (in water pipes/tanks) on organic compounds diffusing into non chemically resistant pipes. e.g. Pseudomonas sp. Air conditioning systems and stagnant water Stagnant water in pipes and air conditioning systems at risk from Legionella. in pipes/tanks Agricultural land and water courses Risks from pathogens in food storage (Aspergillus sp.), slurry, bovine urine, rats and water course (Cryptosporidium parvum and Leptospirosis). 6
  • 7. Pathogens and Land Contamination Examples of Pathogens – Bacillus anthracis • Bacillus anthracis causes the potentially fatal disease anthrax. • It is a zoonotic pathogen and predominantly infects herbivorous animals. • Humans can be infected, but are relatively resistant. Lungs of patient after 4 months of anthrax infection in the lung. Source: Public Health Image Library (PHIL) Dermal Anthrax lesion, source: Public Health Image Library (PHIL) 7
  • 8. Pathogens and Land Contamination Anthrax Infections how do they Occur 2. Germination and multiplication of anthrax in lymphatic system and spleen. Vegetative forms released in large numbers into blood in final hours of life. 3. Vegetative Spores, released at death as haemorrhagic 1. Cow ingests soil exudates from nose, mouth or anus in spilt blood Spores - Sporulate on exposure to oxygen Anthrax spores from previous anthrax deaths or buried infection of further soil infected carcass. 8
  • 9. Pathogens and Land Contamination Anthrax Infections how do they Occur 1. Pulmonary anthrax, inhalation of spore laden dust from the site, entering the lungs. Source: Public Health Image Library (PHIL) 2. Cutaneous infection via a lesion 3. Gastrointestinal, only caused by eating or cut in the skin. infected meat or contaminated water. Has not occurred in the UK in past 100 years Anthrax spores from previous anthrax deaths or buried infected carcass. 9
  • 10. Pathogens and Land Contamination Prevalence of Bacillus anthracis in the UK Anthrax outbreaks in UK livestock: 1887 to 2006 (Source DEFRA) 1600 1400 1200 Number of outbreaks 1000 800 600 400 200 0 1887 1897 1907 1917 1927 1937 1947 1957 1967 1977 1987 1997 • Anthrax was more prevalent in humans in the UK due to a greater incidence in livestock and also greater workplace exposure (e.g. woollen mills). • The number of human cases has dropped steadily over the years, with very few cases since the 1970’s, workers in key industries vaccinated since 1965. 10
  • 11. Pathogens and Land Contamination General routes of infection for Anthrax Most new cases Gardens of anthrax from imported Humans Animal feeds materials Bonemeal Animals Imports Leather Hides, work, Wool, Hair tanneries, Land or water wool Process processing. wastes etc. Industrial Cases can occur Anthrax from historic anthrax Historic contamination contamination 11
  • 12. Pathogens and Land Contamination Anthrax Infections • Today anthrax occurs sporadically with isolated cases reported in humans. • Cases have usually been dermal anthrax. Black boil-like lesions develop on the skin, readily diagnosed and easily treated. • Rarely inhalation anthrax occurs, resulting in haemorrhagic pleural effusions, severe septicaemia, meningitis and high death rates. Recent case caused by imported hides contaminated with anthrax spores, used make into drums. • In 2009-2010 a number of cases of death from anthrax occurred to the intravenous injection of Heroin cut with anthrax. 12
  • 13. Pathogens and Land Contamination Viral Infection - Variola Major Virus • Smallpox is caused by Variola major virus, which has since been eliminated from the world. • There is no treatment and it has high mortality rates. • The last outbreaks were mainly in the 1930s, some sporadic cases until the 1970’s but none afterwards. • Smallpox can survive for long periods of time in dry scabs (13 years has been documented). • In normal conditions, it is unlikely to survive for more than 48 hours. • Intact virus was recovered from an exhumed body (Spitalfields 1985), over 100 years old. • The virus could not be grown in the laboratory and not considered infective. • The risk that smallpox might re-emerge if the remains of smallpox victims are disturbed is remote, but Smallpox infection. Source: Public Health Image Library (PHIL) clearly the impact would be very significant. 13
  • 14. Pathogens and Land Contamination Leptospirosis – Weils Disease • Leptospirosis, a bacterial disease passed from animals to humans (zoonoses). Leptospirosis cell within human kidney. • It can be caught from: Source: Public Health Image Library (PHIL) • work on farms or handling animals (e.g. cows and rats); • contact with canals, rivers and other watercourses; and • work in drainage ditches and sewers; • Infection via contact with infected animal (particularly rats) urine, fluids, tissue or indirect exposure through water, soil or animal foodstuffs. • The bacteria enter the body through cuts in the skin, the skin lining the mouth and nose, and membrane at the front of the eye. • Treated early the prognosis is good, if untreated it can cause liver and kidney failure and be fatal. 14
  • 15. Pathogens and Land Contamination Assessing the Risk – What is Acceptable? • Pathogens are present throughout the environment, but there are different risks posed by different pathogens. • Humans assess the risks posed by pathogens everyday: • Should we go into work when our colleagues have a cold? • Gauging the cleanliness of eating establishments? • Wash your hands after using the toilet? • What is the water quality at a beach? Source: Public Health Image Library (PHIL) • Your bodies immune system is constantly battling against potential pathogens. • When dealing with soils/buildings or water, the risks have to be taken in the context of the threat posed by the likely pathogens and their abundance. • e.g. Anthrax – Highly dangerous and major economic threat to agriculture. No risk is acceptable as only one organism may be required to cause infection; no detectable viable spores/cells is threshold. • e.g. Leptospirosis/Clostridium – potentially abundant in environment, minimise exposure to sensitive receptors as it is impossible to eradicate pathogen. 15
  • 16. Pathogens and Land Contamination The Unique Problem of Pathogens Carcinogenic effect on cells Acutely toxic Pathogenic Rapid substance, Bacterial division of the response cell division cancer cells relates to dose One cell divides into two, two cells into four, four cells into eight, this process continues leading to the growth millions of cells. This is analogous to cancer and the effect of carcinogens. 16
  • 17. Pathogens and Land Contamination Viral Infection and Replication 2. Virus penetrates cell membrane an injects nucleic acid (DNA and RNA) into cell. Coated vesicle 1. Viruses attach to cell Endosome mRNA Translation nucleic acid (DNA and RNA) Nucleus Synthesis of envelope proteins Endoplasmic reticulum Golgi apparatus 4. New viral nucleic acids are 3. Viral nucleic acid packaged into viral particles and replicates using host released from the host cell, which cellular machinery. may be destroyed in the process. 17
  • 18. Pathogens and Land Contamination Risk Assessment – Sources of Contamination • Industrial Sites • The areas of sites that processed animals or products made from animals, posing the greatest risk of pathogen contamination are: • Areas where carcasses or hides/wool were received and stored; • Areas for the washing or early processing of the materials and associated drainage systems; or • Areas used for waste disposal, animal burial or where land raising has taken place. • Areas inhabited by pests (e.g. rodents) which are vectors of disease (applies below) • Targeting these areas would be the most effective approach to detecting pathogens, if free of pathogens it is likely that others are also uninfected. • Agricultural Sites • Does information suggests previous disease outbreaks or animal burials on site, “Animal Health and Veterinary Laboratories Agency” should be contacted. • If burial sites (due to notifiable diseases) are recorded it is unlikely that investigation/development of these areas would be allowed. • Has blood, tissues or anaerobic digester waste been spread on the site. 18
  • 19. Managing Risks from Gas Industry Construction in Contaminated Land What/Who Might be at Risk Surface Water Drainage Inhalation, ingestion or dermal contact of Surface Public/neighbours animalslivestock soils/dusts, containing microorganisms or Water The very young spores. Courses The elderly The ill Building Construction occupants workers The Old Tannery Buried Animals Groundwater Leaching of Microorganism and spores into, drains, surface water and groundwater. 19
  • 20. Pathogens and Land Contamination Risks from Investigation and Construction • The risks from pathogens may be overlooked through ignorance or because they are deemed outside the scope of a project (e.g. Part 2a). • Risks may be higher on sites which are not potentially polluted by chemical pollutants and protection not offered by PPE. • Health and Safety issues which may be overlooked include: • Not including the analysis of pathogens; • Not identifying the risk of potential pathogens; • Adopting suitable drilling methods that minimise waste, aerosols, and dust generation; • Provision of suitable PPE including, the covering of cuts and respiratory protection; • The safe handling, storage and disposal of waste materials; • Quarantining samples until cleared of expected pathogens; and • Good site hygiene practices. 20
  • 21. Pathogens and Land Contamination Assessing the Risk – Other Considerations • The three main routes of infection from pathogens are: dermal contact, inhalation and ingestion. • However, the main route of infection varies between different pathogens; the severity of the disease may also change dependant on the route of infection. • Zoonotic pathogens cause a greater risk to their target host animals, although they can still cause severe disease in humans. • Children are at the greatest risk of infection and serious illness from environmentally transmitted pathogens because: • Absent or poor sanitation habits and more likely to ingest soil; • Immature immune system; and • Intestinal mucosa more permeable to water and reduced stomach acid and pepsin secretion • The elderly and immuno-compromised are also at greater risk from pathogens. • Pathogens may also migrate from the soil into the drains or underlying aquifer; quantifying these risks is also problematic. 21
  • 22. Pathogens and Land Contamination Assessing the risk – Scottish Borders Case • This zero risk approach was taken following a confirmed case of inhalation anthrax in the Scottish Borders. • The source of the anthrax had been traced to a drum made from an imported infected hide. • Some sites where this drum had been used were later found to be contaminated with anthrax spores. • Affected sites were decontaminated until no viable spores were detected. • This ensured there was no risk public health for these sites, albeit at a cost of £460k (£227k - decontamination contract). • Estimates of the human LD50 for inhalation anthrax range from 4,100-10,000 spores, other studies have shown humans can be exposed to around 1,000 spores without contracting anthrax. • Despite the growing evidence, there are still gaps in the dose response relationships between anthrax exposure and infection, especially for dermal infections. • If this information is provided then a risk based approach may be possible. • But one cell is theoretically all that is required for infection. 22
  • 23. Pathogens and Land Contamination Pathogens and Legislation • Planning (PPS 23: Planning and Pollution Control, England) • PPS23 describes the hazards posed by a contaminant to include biological (pathogens) and therefore sites at risk from pathogens proceeding through the planning system requires an investigation to consider the risks posed by pathogens during and after development. • Environmental Liabilities Directive and Environmental Damage Regulations • The definition of environmental damage in respect of 'land damage' is "any land contamination that creates a significant risk of human health being adversely affected as a result of the direct or indirect introduction, in, on or under land, of substances, preparations, organisms or micro- organisms". • The Control of Substances Hazardous to Health (2002) Regulations • COSHH requires that any risk associated with biological agents must be considered prior to worker activity that might place employees or the public at risk; this includes the investigation and remediation of potentially contaminated land. • Pathogens included in EU/UK waste legislation, Hazard H9 - “Infectious” • Pathogens can be dealt with through Statutory Nuisance legislation. 23
  • 24. Pathogens and Land Contamination Pathogens and Legislation • Public Health (Control of Disease) Act 1984, (substantially amended by the Health and Social Care Act 2008) and various Health Protection Regulations 2010, including the Health Protection (Part 2A Orders) Regulations 2010: • Local authorities and JP to take action to protect public health from a risk of significant harm from infection or contamination. Registered medical practitioners and laboratories, to notify a specific list of notifiable infectious diseases and causative agents and also other infections or contamination, such as with chemicals or radiation, that may pose a significant risk to public health. • Specified Animal Pathogens Order 2008 (SAPO), A person must immediately notify a veterinary inspector if: • has possession of any thing in respect of which they have reasonable grounds for suspecting that an animal pathogen specified in Part 1 of Schedule 1 (e.g. B.anthracis) is present and does not have a licence in respect of that pathogen. • Notifiable diseases in animals must be reported to the Animal Health and Veterinary Laboratories Agency. • Notifiable diseases in humans must be reported to the local Consultant in Communicable Disease Control and under RIDDOR. • Pathogen specific legislation, e.g. The Anthrax Order 1991 24
  • 25. Pathogens and Land Contamination Part 2A and Pathogens • Pathogens are not included or excluded from the definition of a “substance”. • A substance is defined in section 78A(9) of Part 2A of the Environmental Protection Act 1990 as: “any natural or artificial substance, whether in solid or liquid form or in the form of a gas or vapour”. • Can a pathogen be classed as a ‘substance’ under Part 2A? - the definition of a substance is unspecific and unclear; and - the definition of a substance in Part 2A has not been looked at in case law. • The position of the Environment Agency was that microorganisms are not mentioned in the definition of a “substance” in Part 2A, whereas in other legislation they are. • They believe that their omission from Part 2A was intended and that pathogens should not be considered a “substance” under Part 2A, and therefore should fall outside the remit of Part 2A. • However – the decision lies with the local authority, only they are able to determine a site to be contaminated land based on the presence of pathogens under Part 2A! 25
  • 26. Pathogens and Land Contamination Analysis of Pathogens • Tests for many pathogens have been developed for detection from human or animal bodily fluids; they are not optimised for detection from soils/water. • Some tests used for detection in soil/water samples do not accurately quantify pathogen numbers (cells/spores/viruses). • With the exception of Legionella, there are a limited number of laboratories that can test for pathogens from environmental samples. • Seek advice from experienced microbiologists. • Tests must be carried out in specialist microbiology facilities; B. anthracis requires Level 3 Containment. • Analytical methods are based one or both of: • culturing, growing cells in broth or agar plates, or growing virus in tissue culture • molecular detection, analysis of RNA or DNA. 26
  • 27. Pathogens and Land Contamination Analysis of Pathogens • For the rapid selective isolation and detection of Bacillus anthracis in potentially contaminated materials (e.g. animal hair or soil) the HSL uses a combination of enrichment culture and molecular detection by polymerase chain reaction (PCR). • Culture takes overnight and PCR, which tests for unique DNA sequences from the B. anthracis toxin genes, takes 4-6 hours. • Similar approaches can be used for the detection of spore forming Clostridium Sp. and opportunistic pathogens such as Pseudomonas Sp. in soils, water and building material. • Legionella testing in water is commonly undertaken by commercial environmental Labs; this is a culture based test. • Leptospira can be analysed in water samples although it is a lengthy process (6 weeks) and not recommended for large surface water bodies. It is feasible to undertake PCR based tests. Testing in humans is done by the HPA in Hereford. • The detection of Smallpox in humans could only be undertaken at a specialist facility such as (HPA Porton Down) 27
  • 28. Pathogens and Land Contamination Factors to Consider for PCR Testing • Molecular detection by PCR is rapid (hours), sensitive (our anthrax PCR detects down to less than 10 spores) and specific….. BUT: Source: Public Health Image Library (PHIL) • Molecular tests can be affected by interferents such as humic acids and organic debris – important for environmental/soil samples to have a validated clean-up step. • A positive result shows that DNA sequences are present, not necessarily live micro- organisms. Which is why we use culture as a back-up. However a rapid PCR positive result can be used as an alert for countermeasures or to focus further tests. • The assay results are only as good as the sample taken. 28
  • 29. Pathogens and Land Contamination Sampling Strategies • Statistically based strategies: • For example, Visual Sample Plan (VSP) – Pacific Northwest National Laboratory and US Dept of Energy http://vsp.pnnl.gov/ - a software tool for optimal sampling design and statistical analysis for environmental monitoring, characterisation and remediation, response to CBRN events, sampling of soils, buildings, groundwater, sediment, surface water, subsurface layers. • Favored by Government Decontamination Service for sampling to support clean up. • Targeted/intelligence led sampling: • Be prepared to step outside the Plan-led sample collection – in the Scottish anthrax case the main focus of contamination was one floor mat. Use your eyes and imagination. • Supplement statistical sample plan with site specific information. • Details of past history and use of site. Talk to ex employees or residents. • Previous location of buildings, activities, effluent drains 29
  • 30. Pathogens and Land Contamination Bioaerosol Sampling • Examples of fixed point and personal sampling methods for measuring bioaerosol emissions: • Andersen impactors; Liquid impingers; • 30
  • 31. Pathogens and Land Contamination Bioaerosol Sampling • Filtrations samplers (fixed point and personal): 31
  • 32. Pathogens and Land Contamination Worker Protection • Levels of live pathogens are likely to be low. • General personal protection for outdoor work, including gloves, covering cuts and grazes and where required respiratory protection, likely to provide sufficient protection. • During building remediation or demolition, asbestos protection also sufficient protection against pathogens (with safe disposal of contaminated PPE). • This should be supplemented by a procedure for recording exposures, injuries and any subsequent ill-health. • Suitable secure containers for storage of samples and transportation to analyst. • Adequate information accompanying samples to ensure safe handling for analysts. 32
  • 33. Pathogens and Land Contamination Future Considerations • Continued emergence of non-fossil fuel energy technologies. • Examples include mechanical & biological treatment (MBT), waste or biomass to energy, anaerobic digestion (AD). • Potential for biological and chemical residues from the process contaminating surrounding soil, leachate run-off, bioaerosol emissions, also asphyxiant gases in confined spaces. • Also subsequent spreading of wastes, such as AD waste, to land needs to be managed carefully. • Scaled-down waste to energy projects (e.g., community heat and power) may be less well managed. 33
  • 34. Pathogens and Land Contamination Thanks for your Attention Dr Russell Thomas, Technical Director Parsons Brinckerhoff Ltd thomasru@pbworld.com Tel: 0117 9339262 Dr Brian Crook, Microbiology Team Leader Health and Safety Laboratory brian.crook@hsl.gov.uk Tel: 01298 218421