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BIOLOGICAL MONITORING OF WORKERS
                        EXPOSED TO PESTICIDES
                Guidelines for application in field settings




        Photo by Leslie London
        University of Cape Town




        Occupational Health Research Unit Monograph

        Department of Community Health

        University of Cape Town Medical School

        Anzio Road

        Observatory

        7925

        Original work from 21 September 1999


Leslie London, University of Cape Town
Biological Monitoring of Workers Exposed to Pesticides by Leslie London, Centre for Occupational and Environmental
                   Health Research, Health Sciences, University of Cape Town is licensed under a

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                                                         June 2011




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Leslie London, University of Cape Town
                                                                                                                     2
PREFACE
       This guideline was produced for those persons responsible for the maintenance of health and safety
       measures at agricultural workplaces handling potentially hazardous organophosphate and carbamate
       chemicals. It is primarily aimed at professional nursing and other medical staff charged with monitoring
       workers for pesticide exposure, but will be useful to all personnel involved in workplace health and safety
       wishing to understand the principles behind monitoring workers for pesticide exposure.

       The guidelines concentrate on monitoring for organophosphate and carbamate insecticides because the
       technology is reasonably readily available, and the methodology well described. These chemicals are also
       widely used, and are the most common cause of acute poisoning by pesticides. The guidelines have also
       been written bearing in mind the Hazardous Chemical Regulations (Regulation 5549 of 25 Aug 1995 in terms
       of the Occupational Health and Safety Act) that include agricultural workplaces in addition to industry.

       The information contained in these guidelines is based on the most current thinking and published research
       on the topic, and a relevant bibliography is included at the back for those interested in reading further.

       Abbreviated summaries of the recommended protocols are contained at the end of this document.

       This guideline has been produced by the Occupational and Environmental Health Research Unit of the
       Department of Community Health, University of Cape Town as part of its research in the field of pesticide
       hazards and pesticide safety. The support of the International Development Research Centre (IDRC) in this
       regard is acknowledged.

       Further copies may be ordered from the Unit on request. Contact Anouchka at email nouch@anat.uct.ac.za
       or fax 021 4066163 for more information.

       Leslie London

       Occupational and Environmental Health Research Unit

       2nd edition (21 September 1999)



     Disclaimer note: Please note that this information was correct at the time of writing, but since then some
          information may be out-dated. For suggestions or updates, please contact healthoer@uct.ac.za




Leslie London, University of Cape Town
                                                                                                                    3
CONTENTS                                                                               Page

   1. Why monitor workers exposed to pesticides                                        5

   2. What type of monitoring is available?                                            6

   3. What chemicals may be monitored                                                  7

   4. Biological monitoring of exposure to Organophosphate and carbamate pesticides    8

   5. How may one know whether a cholinesterase level is low, or has dropped?          9

   6. How does one establish a baseline cholinesterase level?                          10

   7. How much of a decrease in cholinesterase is significant?                         10

   8. What type of preventive action should be prompted by a drop in cholinesterase?   11

   9. How frequently should testing be done?                                           12

   10. What technology is available for testing in the field?                          12

   11. How should a monitoring programme be managed?                                   13

   12. What are the common pitfalls in implementing biological monitoring programmes

       for organophosphate and carbamate pesticides?                                   14

   13. How should monitoring programmes be evaluated?                                  14

   14. A checklist before implementing a monitoring programme                          15

   15. Diagnostic tests compared to Biological Monitoring                              15


       Summary protocol for a biological monitoring programme
       for workers exposed to organophosphate carbamate pesticides                     17




Leslie London, University of Cape Town
                                                                                              4
BIOLOGICAL MONITORING OF WORKERS EXPOSED TO PESTICIDES
                                            A Guideline for field application


             1.                    Why monitor workers exposed to pesticides?

             Pesticides are chemicals designed to have adverse effects on various plant and insect species. As a
             result, they may have unintended adverse effects on humans and the environment. Workers
             involved in the manufacture,        formulation, preparation, packaging, transport, storage, mixing and
             application of pesticides will have the      highest exposures to pesticides and therefore have the
             highest health risks. Exposures may be aggravated by        handling pesticides in a closed room.
             Absorption of pesticides through the skin is a very important route of      exposure, and wet overalls
             may be a significant hazard.

             In order to protect workers who are exposed to pesticides from adverse health effects, monitoring of
             workers may be performed to detect early biochemical or physiological changes before these lead to
             reversible or irreversible disease and illness. A simplified model is depicted below to illustrate the use
             of monitoring. The primary purpose of monitoring is therefore to prevent pesticide-related disease by
             detecting early changes before the exposure causes frank disease.


                              Figure Model for the development of chemical-related disease



                    EXPOSURE TO               EARLY BIOCHEMICAL OR
                                                                          REVERSIBLE DISEASE        IRREVERSBLE
                    CHEMICAL                  PHYSIOLOGICAL CHANGES
                                                                                                    DISEASE




                  From Leslie London
                  University of Cape Town


                  A secondary function of monitoring may be to detect the presence of disease amongst exposed
                  workers (diagnostic testing or screening) for purposes of preventing further deterioration, treating
                  the disease, securing compensation or establishing a long-term prognosis. However, these aspects
                  will not be covered here as the aim of this monograph is to provide guidelines as to how pesticide-
                  related disease may be prevented by the monitoring of workers exposed to pesticides.




Leslie London, University of Cape Town
                                                                                                                          5
2.                What type of monitoring is available?

             Broadly speaking, there are two main methods of monitoring exposures to potentially hazardous
             chemicals

              a)       Environmental monitoring of the chemical or its residue in the environment (air, foliage, soil)
                      or in contact with humans (overalls, skin contact)
              b)       Monitoring of the intact chemical or its metabolite in the tissue or fluids of the body
                      (Biological Monitoring), or the effect of the chemical on enzyme systems within the human
                      body (Biological Effect monitoring)

              Because there are many variables that determine whether a chemical will be absorbed from the skin
              or through the lungs into the human body (use of protective equipment, safety practices climatic
              conditions, individual susceptibility, concomitant disease, properties of the formulation and the
              chemical. etc), biological monitoring is regarded as a more accurate assessment of human exposure.
              Once the intact pesticide is absorbed, it may be metabolised, redistributed in the body and
              differentially deposited within body tissues. For this reason the measurement of the biological effect
              of a pesticide within the body is regarded as a better indicator of exposure to a pesticide. The
              sequence of steps in chemical toxicity is illustrated in the figure on the next page.




                                                                           From Leslie London
                                                                           University of Cape Town




Leslie London, University of Cape Town
                                                                                                                     6
Figure: Chemical Toxicity in Humans

                                            EXPOSURE TO CHEMICAL



                                         DEPOSITED ON SKIN OR IN LUNG



                                                ABSORPTION

                                                 METABOLISM

                                                REDISTRIBUTED

                                                DEPOSITED IN
                                                TISSUES



                            EFFECTIVE               EFFECTIVE                  EFFECTIVE
                            TISSUE DOSE ₁           TISSUE DOSE ₂              TISSUE DOSE ₃



                            BIOLOGICAL                BIOLOGICAL               BIOLOGICAL
                            EFFECT ₁                  EFFECT ₂                 EFFECT ₃




          From Leslie London
          University of Cape Town                                                    DISEASE

               3.         What chemicals may be monitored?

               For most organophosphate and carbamate pesticides, a fairly simple measure of biological effect is
               available. This is the measurement of the enzyme cholinesterase, which is inhibited by the
               carbamate and organophosphate pesticides. The enzyme cholinesterase may be measured in the
               blood (either within the red blood cells, or in the blood plasma surrounding the blood cells).
               Lowered levels of cholinesterase activity indicate exposure to organophosphate or carbamate
               pesticides. The mechanism by which these pesticides cause adverse effects is by inhibiting the
               cholinesterase in the nervous system. By measuring the cholinesterase in the blood, one can
               determine the activity of the chemical in the body before it has an effect on the nervous system.




Leslie London, University of Cape Town
                                                                                                                    7
For most other pesticides, such simple methods of biological effect monitoring are generally not
            available. lf one wishes to monitor workers for exposure to pesticides other than organophosphates
            and carbamates, the tests required are complex, time-consuming and costly (involving measurement of
            the intact pesticide or its metabolite in the blood or urine of the exposed worker, or the intact
             pesticide in the environment). These tests are generally not widely available in South Africa at
             commercially practicable costs despite their requirement in terms of the HCS regulations.

            This monograph concentrates on biological monitoring of exposure to organophosphate and carbamate
            pesticides. However, it is recommended that where other highly toxic agents are in use (e.g.
            pentachlorophernol) every attempt should be made to establish a monitoring programme for these
            agents.


            4.           Biological monitoring of exposure to organophosphate and carbamate pesticides



           Organophosphates and carbamates bind to the enzyme cholinesterase in the human nervous system,
           causing an accumulation of chemical neurotransmitters. This, in turn, leads to an overactivity within the
           nervous system manifesting as the symptoms of acute poisoning. By measuring the effect of these
           pesticides on similar enzymes in the blood, one can detect a decrease in enzyme activity before
           symptoms develop and therefore prevent poisoning

           Of the two blood cholinesterases, plasma cholinesterase is regarded as most sensitive to recent
           absorption of Ops and carbamates, while the red blood cell cholinesterase reflects more closely the
           concurrent effect in the nervous system. Thus both enzymes may be used to prevent nervous system
           poisoning, but different action levels apply to the two enzymes. This is dealt with in sections 7, 8 and 9.

           If a monitoring programme is to be used it is preferable that both enzymes are monitored. If financial
           resources are limited, the red cell cholinesterase is preferred as the best assay because it reflects more
           closely the physiological effect of the chemicals on the worker’s nervous system.




Leslie London, University of Cape Town
                                                                                                                         8
5.                   How may one know whether a cholinesterase level is low, or has dropped?

        Because cholinesterase levels are known to differ widely between individuals, irrespective of their exposure
        to chemicals, the optimal method of determining what is normal for a person is to establish the individual’s
        baseline level. This can be compared to enzyme levels after exposure to organophosphates or carbamates
in
        order to determine whether a drop has occurred. The figure below illustrates a hypothetical case where
        serial testing for cholinesterase level is performed before, during and after exposure. By withdrawing the
        worker from exposure timeously, he or she may be prevented from becoming symptomatic, and may safely
        be returned to work once their cholinesterase has returned to normal.

                                   Figure: Serial testing for cholinesterase before, during
                                   and after exposure organophosphates or carbamates




          From Leslie London
              University of Cape Town

        In the absence of a baseline for an individual worker, it is difficult to determine whether a workers
        cholinesterase has decreased. Laboratories often quote normal ranges for cholinesterase levels, but
        because the variation between individuals is so high, these 'normal' ranges are not helpful for biological
        monitoring. For example, a few workers who have no significant exposures of organophosphates or
        carbamates may have cholinesterase levels well below the 'normal' ranges simple because of genetic
        variability. The normal ranges developed for laboratory use are generally based on statistical distributions,




Leslie London, University of Cape Town
                                                                                                                        9
and bear little relationship to the practical distributions, and bear little relationship to the practical
           applications of biological monitoring. For this reason, it is advisable to develop individual baselines for
           each worker in the monitoring programme, rather than rely on normal ranges to identity exposed
           workers.


           6.         How does one establish e baseline cholinesterase level?

           To be sure that it is a baseline, the person should preferably be tested before starting work. Alternatively,
           if already in employ, he or she could be tested in the course of the year as long as they have had no
           exposures to organophosphates or carbamates within the previous 2 or 3 months. This is the time taken
          for the red blood cell cholinesterase to recover alter significant exposure. It is therefore important that
          the worker is not tested tor a baseline while his/her cholinesterase is still below their true normal.


          Besides the natural differences in cholinesterase levels between people, there are also biological
          fluctuations from one day to another. While these differences are small, it may still affect the level at
          which you establish the individual’s baseline level, especially if the precision of the method used to
          measure cholinesterase is low. For this reason, it is better to take the mean of two measurements to
          establish a baseline level of cholinesterase for subsequent comparisons.


          7.         How much of a decrease in cholinesterase is significant?

          Researchers have correlated the level of decline in cholinesterase with the development of symptoms in a
          series of studies. It is generally agreed that a decline at 40% or more in plasma cholinesterase (i.e. down
          to 60% of baseline levels or lower) is associated with symptoms, while for red blood cell cholinesterase
          the decline required to produce symptoms is 30% or more (i.e. down to 70% of baseline levels or lower).
          [Note the HCS regulations cite a drop to 70% of baseline levels of red cell cholinesterase as the Biological
          Exposure Index for exposed workers.]

          In order to prevent the onset of symptomatic disease, declines of less than those described above should
          warrant preventive actions, which may include investigating the work environment, re-testing the worker
          or removing the workers from any further exposure.

          However, because individuals' cholinesterase levels differ slightly from day to day, it is important to be
          able to distinguish a benign daily fluctuation from a significant decline warranting further action.
          Research has shown that biological fluctuations should not exceed 10 to 15% from day to day. For this
          reason, declines of between 10 and 30% (red cell cholinesterase) and between 10 and 50% (plasma
          cholinesterase) are significant declines that could be used to prompt preventive action.




Leslie London, University of Cape Town
                                                                                                                         10
8.        What type of preventive action should be prompted by a drop in cholinesterase?

           The table below is based on regulations of the California Health Department and lists the preventive
           actions that should be taken at different levels of cholinesterase decline from baseline.

           If a worker has been removed from exposure as a result of a decline in cholinesterase, he or she should
           not return to work until the red blood cell cholinesterase has risen to at least 90% of the baseline value
           (i.e. within normal variability). It is important to have alternative work available for these workers while
           their cholinesterase levels are depressed.

           When a worker’s baseline cholinesterase is based on two readings, its precision is increased and it will be
           easier to tell whether a small decline in cholinesterase is significant or not.




Leslie London, University of Cape Town
                                                                                                                      11
9.         How frequently should testing be done?

            Once a baseline is established for a worker, it is advisable that he or she be regularly tested during
            ongoing exposure. Testing should happen at least once during the spray season, preferably around
            peak spraying time. However, it is advisable to test more regularly than this. For example, California
            regulations prescribe that workers having more than 6 full days exposure to organophosphates or
            carbamates within a 30 day cycle, should be tested, or that a worker with more than 40 hours contact
            with pesticides in a weeks' schedule should be tested. Other factors particular to the work setting may
            also be used to prompt testing, such as continuous work with pesticides in a closed environment, or
            known accidental exposure. Note that the HCS regulations leave the choice of the timing and frequency
            of testing to the discretion of the occupational health practitioner.


            10.        What technology is available for testing in the field?

            Usually, cholinesterase testing is available from commercial and University laboratories at competitive
            rates. This requires the presence of professional staff available to draw blood under ice to the relevant
            laboratory, and awaiting the laboratory result. One drawback is the potential delay between the taking
            of the blood and receipt of the result, or, worse still, loss of the sample of the result in transit.

            An additional consideration is the need to ensure that the laboratories practice adequate quality
            control of cholinesterase estimation. This is particularly the case for the red cell cholinesterase assay,
            for which the methodology is fairly complex and susceptible to many sources of error. For this reason,
            it would be advisable to ensure that the laboratory to which venous blood samples are sent, can give
            reasonable account of their efforts to ensure quality control, or, preferably, demonstrate that they are
            part of laboratory quality control programme for the tests in question. Such a programme has been
            suggested by the National Centre for Occupational Health in Johannesburg.

             An alternative technology for cholinesterase estimation involved field kits based on finger prick devices.
             A number of such devices are available, although their reliability and validity are not widely described.
             One such field kit (the TestMAte OP) has been tested under field and non-field conditions in the
            Western Cape and shown to have sufficient reliability as to be able to apply the California regulations
            with reasonable robustness. Such technology may be easily applied by field staff (not necessarily health
            professionals) with sufficient training, and the benefits of immediate results in the field may outweigh
            the slight loss of precision involved.




Leslie London, University of Cape Town
                                                                                                                    12
11.          How should a monitoring programme be managed?

                Clearly, the decisions as to who will be responsible for the planning, implementation and evaluation
                of the programme will lie within the ambit of the workplace organisation. However, certain
                questions will need to be decided by those responsible for the programme. These would include:

                a.    What workers will be included in the monitoring programme?

               This will be informed by your risk assessment, required of employers in terms of the HCS
               Regulations. It is advisable to include all workers who handle pesticides, whether they are involved
               in the mixing, application, storage or transport of pesticides in a monitoring programme. Other
               specific indications may be added depending on specific conditions - e.g: Workers who perform
               maintenance work on spray equipment, or who live in dwellings exposed to spray, etc. Risk
               assessments to identify the presence of exposures and the workers at risk should be conducted at
               least every two years, but preferably more frequently.

               b.     How often should testing be done?

               As indicated above, a baseline should be performed followed by periodic testing, at least once
               per season, but preferably more often, timed in relation to intense spraying activities.

               c.     What thresholds should prompt preventive actions?

               Thresholds that should prompt preventive actions are summarised in the table above.

               d.     What follow up is needed?

               Affected workers must be followed up until their cholinesterases have returned to normal.
               Investigation of the work place must be done to identify behavioural, structural and organisational
               sources of exposure amenable to intervention. Appropriate training and hygiene measures should
               then follow.

               e.     How long should records be kept?

               Record keeping is an essential component of good industrial hygiene practice as well as comprising
               an important requirement of the imminent Hazardous Chemical Substance Regulations under the
               Occupational Health and Safety Act. In terms of these regulations, all workplaces where
               organophosphate chemicals are used will be required to perform appropriate medical monitoring of
               exposed workers, and to be able to demonstrate evidence of such monitoring to the department of
               Labour inspectorate. The regulations insist on records being kept for at least 30 years.




Leslie London, University of Cape Town
                                                                                                                  13
Additional reasons for maintaining adequate records are the need for proper documentation for
             potential compensation requirements, and the potential for many of the cholinesterase compounds to
             cause long-term chronic health effects. Good monitoring data during the course of employment will be
             essential in interpreting chronic neurological and other disorders arising in workers, as well as in
             Assisting future research into many of the long-term hazards of ongoing chemical exposures.

             f.    Informed consent

             It is important to remember that a monitoring programme involves invasive procedures (the taking of
             blood) and therefore workers should only participate after giving full informed consent. This is an
             additional argument for having monitoring programmes supplemented by education and training as
             part of a comprehensive health and safety programme.

             12.    What are the common pitfalls in implementing biological monitoring programmes for
                    organophosphate and carbamate pesticides?

             Biological monitoring will provide a useful means preventing disease due to exposure to
             organophosphate and carbamates. However, it cannot serve as substitute for other methods of
             pesticide safety, such as engineering controls, correct use of personal protective equipment,
             administrative controls and worker education and training, all of which should be integrated in a
             comprehensive health and safety progamme at the workplace, and which are feasible in the
             agricultural setting.

             Once a monitoring system is in place, it should be regularly evaluated to ensure that it is effective
             and efficient. Too often, programmes run routinely to meet statutory requirements without
             attention to whether they are achieving what they set out to achieve.

             Quality control is often neglected in many monitoring programmes, particularly with regard to
             laboratory results. An efficient programme can be a waste of time and resources if data from the
             laboratories is invalid. You should insist on evidence of adequate quality control from the
             laboratories where you have conducting your analyses. If necessary, submit one or more
             duplicate samples from the same subject with an alias/aliases. By comparing the results, you will
             be able to get e good quantitative idea of the laboratory’s precision.

             13.     How should monitoring programmes be evaluated?

             Depending on the scope of the programme, its objectives and the amount of resources available,
             evaluation of a monitoring programme could examine different aspects.




Leslie London, University of Cape Town
                                                                                                                     14
Firstly, the programme needs to establish that it is achieving the prevention of disease due to
               organophosphates and carbamate pesticides. Surveillance foe cases of poisonings reported at work
               and outside the workplace involving exposed workers can provide this type of information. This is a
               measurement of the effectiveness of the programme. Collaboration between farm health personnel
               and information flow in order to achieve this objective.

               Secondly, the process outputs of the programme should be evaluated. How many workers were
               monitored, how many tests were done and how many preventive actions (retesting, workplace
               investigations, withdrawal from exposure, etc) were triggered by the programme? This is an
               indication of how many workers were protected from potential disease by the application of the
               programme.

               Thirdly, how efficient was the programme implemented? Were tests performed in the correct
               manner, and at the appropriate times? Were the correct workers monitored? Were there any
               important omissions from the programme?

               Fourthly, what were the costs, direct and indirect, and what were the benefits in financial terms?

               Lastly, what was the functional benefit to workplace morale, productivity, occupational health
               service staff job satisfaction etc?

               14.         A checklist before implementing a monitoring programme

               Based on the above, it is helpful to use a checklist of questions before implementing a monitoring
               programme:

                    What are the objectives of the programme? Are they clear?
                    lf prevention is to be the objective of the programme, what protocols will be followed to ensure
                     that the objective can be met? Are these protocols clear to all personnel involved?
                    Will the information recorded enable you to audit the programme?
                    Is the programme linked to an education package?
                    Is the programme going to be sustainable?

               15.         Diagnostic Tests compared to Biological Monitoring

               There are occasions when testing will be used to detect the presence of illness in workers exposed to
               organophosphate and carbamate pesticides. For example, workers presenting to the farm nurse
               with non-specific symptoms such as headache, nausea or dizziness may be suffering from early or
               mild organophosphate poisoning. Under such circumstances, cholinesterase testing may be used to
               diagnose the baseline level is available, comparison of the result will enable the clinician to make a
               simple diagnosis.




Leslie London, University of Cape Town
                                                                                                                    15
However, clinicians are often faced with a situation where no baseline is available, and where the single
       cholinesterase result is within or only slightly below ‘normal’ laboratory range. Under these circumstances,
       serial testing of cholinesterase after withdrawing the worker from exposure may be useful to confirm mild
       cases of poisoning. If the cholinesterase level gradually increases by more than 15 % over time, and then
       plateaus out with a retrospective confirmation of cholinesterase inhibition as the cause of the worker's
       illness. This is clearly of no use in trying to prevent poisoning (the purpose of biological monitoring), but is
       useful in the clinical setting of trying to reach a diagnosis.

       Note that all cases of occupational poisoning including those diagnosed retrospectively must be reported to
       the Chief Inspector: Occupational Health and Safety, Department of Labour in terms of the OHSA. At the
       present time, remember also that any cases of pesticide poisoning are notifiable to the Department of
       Health in terms of the Health Act.




                            From Andrea Rother
                            University of Cape Town




Leslie London, University of Cape Town
                                                                                                                      16
SUMMARY PROTOCOL FOR A BIOLOGICAL MONITORING PROGRAMME FOR WORKERS EXPOSED TO
        ORGANOPHOSPHATE CARBAMATE PESTICIDES

       Planning Monitoring Programme:      Objectives
                                           Who participates
                                           Frequency of testing
                                           Alternative job sites
                                           Responsibilities for investigation
                                           Training
                                           How to evaluate
                                           What technologies to use

       Baseline Cholinesterase Testing:    Pre-employment or at least
                                           2 months without exposure

       Interval testing:       Every month it more than 6 days exposure
                               Per month
                               Other indications

       Depending on level of Cholinesterase:

       Re-test               Investigate       Withdraw from exposure
                                               Wait till CHE back to normal




Leslie London, University of Cape Town
                                                                                        17
Bibliography

       Ames RG, Brown SK, Mengle DC, Kahn E, Stratton JW, Jackson RJ. (1989). Protecting agricultural applicators
       from overexposure to cholinesterase-inhibiting pesticides: Perspectives from the California Programme J.Soc
       Occup Med. 39, 85-92.

       Ames RG, Brown Sk, Mengle DC, Kahn E. (1989b) Cholinesterase activity depression among California
       agricultural pesticide applicators. Am J Ind Med. 15 ; 143-l50.

       Brown SK, Ames RG, Mengle DC. (1989). Occupational Illness from Colinesterase-inhibiting pesticides among
       agricultural applicators in California, 1982-1985. Arch Env Health, 44: 34 - 39.

       Coye MJ, Lowe JA, Maddy KT. (1986a). Biological Monitoring of agricultural workers exposed to pesticides:
       11. Monitoring of intact pesticides and their metabolites. J Occup Med 28(8): 628 - 442.

       Fillimore Cm, Lasenger JE. (1993). A cholinesterase testing programme for pesticides applicators. J Occup
       Med. 35: 61 -70.

       Hulka BS, Margolin Bh. (1992). Methodological Issues in Epidemiological Studies using biologic markers. Int J
       Epidem 135 : 200- 209.

       Krieger Ri, Ross JH. (1993). Risk assessments in the pesticide regulatory process. Ann Occup Hyg. 37: 565 -
       578.

       Lotti, M (1992). Central neurotoxicity and behavioural effects of anticholinesterases. ln: Clinical and
       experimental toxicology of organophosphate and carbamates. Eds Ballantyne B, Marrs TM, Aldridge Wn.
       Butterworth, Stoneham, MA.

       Minton NA, Murray VSG (1988). A review of organophosphate poisoning. Medical Toxicology 3 : 350 - 375.

       Toxicology. Special edition as a Manual on Biological monitoring for Pesticides. 17 June 1994. 19(1).

       Woolen Bh. (1993) Biological monitoring for pesticide exposure. Ann Occup Hyg. 17 : 525 - 540.

       World Health Organisation (1986b). Organophosphorus insecticides : A general introduction Environmental
       Health criteria 63. World Health Organisation, Geneva.




      We would appreciate your feedback for this Open Educational Resource (OER), by completing this form.
                            Alternatively, you can email us at healthoer@uct.ac.za




Leslie London, University of Cape Town
                                                                                                                     18

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Biological Monitoring of Workers Exposed to Pesticides - Guidelines for application in field settings

  • 1. BIOLOGICAL MONITORING OF WORKERS EXPOSED TO PESTICIDES Guidelines for application in field settings Photo by Leslie London University of Cape Town Occupational Health Research Unit Monograph Department of Community Health University of Cape Town Medical School Anzio Road Observatory 7925 Original work from 21 September 1999 Leslie London, University of Cape Town
  • 2. Biological Monitoring of Workers Exposed to Pesticides by Leslie London, Centre for Occupational and Environmental Health Research, Health Sciences, University of Cape Town is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 2.5 South Africa License June 2011 You are free: to Share – to copy, distribute and transmit the work to Remix – to adapt the work Under the following conditions: Attribution. You must at attribute the work in the manner specified by the author or licensor (but not in any way that at suggests that they endorse you or your use of the work) Non-commercial. You m may not use this work for commercial purposes. Share Alike. If you alter, transform, or build upon this work, you may distribute the resulting work but only under the e same or similar license to this one. • For any reuse or distribution, you must make clear to others the license terms of o this work. One way to do this is with a link to the license web page: http://creativecommons.org/licenses/by-nc-sa/ 2.5/za • Any of the above conditions can be waived if you get permission from the copyright holder. • Nothing in this license impairs or restricts the authors’ moral rights. • Nothing in this license impairs or restricts the rights of authors whose work is r referenced in this document. • Cited works used in this document must be cited following usual academic conventions • Citation of this work must follow normal academic conventions Source work available at Vula. Permissions beyond the scope of this license may be available at www.healthedu.uct.ac.za or contact healthoer@uct.ac.za We would appreciate your feedback for this Open Educational Resource (OER), by completing this form. Alternatively, you can email us at healthoer@uct.ac.za Leslie London, University of Cape Town 2
  • 3. PREFACE This guideline was produced for those persons responsible for the maintenance of health and safety measures at agricultural workplaces handling potentially hazardous organophosphate and carbamate chemicals. It is primarily aimed at professional nursing and other medical staff charged with monitoring workers for pesticide exposure, but will be useful to all personnel involved in workplace health and safety wishing to understand the principles behind monitoring workers for pesticide exposure. The guidelines concentrate on monitoring for organophosphate and carbamate insecticides because the technology is reasonably readily available, and the methodology well described. These chemicals are also widely used, and are the most common cause of acute poisoning by pesticides. The guidelines have also been written bearing in mind the Hazardous Chemical Regulations (Regulation 5549 of 25 Aug 1995 in terms of the Occupational Health and Safety Act) that include agricultural workplaces in addition to industry. The information contained in these guidelines is based on the most current thinking and published research on the topic, and a relevant bibliography is included at the back for those interested in reading further. Abbreviated summaries of the recommended protocols are contained at the end of this document. This guideline has been produced by the Occupational and Environmental Health Research Unit of the Department of Community Health, University of Cape Town as part of its research in the field of pesticide hazards and pesticide safety. The support of the International Development Research Centre (IDRC) in this regard is acknowledged. Further copies may be ordered from the Unit on request. Contact Anouchka at email nouch@anat.uct.ac.za or fax 021 4066163 for more information. Leslie London Occupational and Environmental Health Research Unit 2nd edition (21 September 1999) Disclaimer note: Please note that this information was correct at the time of writing, but since then some information may be out-dated. For suggestions or updates, please contact healthoer@uct.ac.za Leslie London, University of Cape Town 3
  • 4. CONTENTS Page 1. Why monitor workers exposed to pesticides 5 2. What type of monitoring is available? 6 3. What chemicals may be monitored 7 4. Biological monitoring of exposure to Organophosphate and carbamate pesticides 8 5. How may one know whether a cholinesterase level is low, or has dropped? 9 6. How does one establish a baseline cholinesterase level? 10 7. How much of a decrease in cholinesterase is significant? 10 8. What type of preventive action should be prompted by a drop in cholinesterase? 11 9. How frequently should testing be done? 12 10. What technology is available for testing in the field? 12 11. How should a monitoring programme be managed? 13 12. What are the common pitfalls in implementing biological monitoring programmes for organophosphate and carbamate pesticides? 14 13. How should monitoring programmes be evaluated? 14 14. A checklist before implementing a monitoring programme 15 15. Diagnostic tests compared to Biological Monitoring 15 Summary protocol for a biological monitoring programme for workers exposed to organophosphate carbamate pesticides 17 Leslie London, University of Cape Town 4
  • 5. BIOLOGICAL MONITORING OF WORKERS EXPOSED TO PESTICIDES A Guideline for field application 1. Why monitor workers exposed to pesticides? Pesticides are chemicals designed to have adverse effects on various plant and insect species. As a result, they may have unintended adverse effects on humans and the environment. Workers involved in the manufacture, formulation, preparation, packaging, transport, storage, mixing and application of pesticides will have the highest exposures to pesticides and therefore have the highest health risks. Exposures may be aggravated by handling pesticides in a closed room. Absorption of pesticides through the skin is a very important route of exposure, and wet overalls may be a significant hazard. In order to protect workers who are exposed to pesticides from adverse health effects, monitoring of workers may be performed to detect early biochemical or physiological changes before these lead to reversible or irreversible disease and illness. A simplified model is depicted below to illustrate the use of monitoring. The primary purpose of monitoring is therefore to prevent pesticide-related disease by detecting early changes before the exposure causes frank disease. Figure Model for the development of chemical-related disease EXPOSURE TO EARLY BIOCHEMICAL OR REVERSIBLE DISEASE IRREVERSBLE CHEMICAL PHYSIOLOGICAL CHANGES DISEASE From Leslie London University of Cape Town A secondary function of monitoring may be to detect the presence of disease amongst exposed workers (diagnostic testing or screening) for purposes of preventing further deterioration, treating the disease, securing compensation or establishing a long-term prognosis. However, these aspects will not be covered here as the aim of this monograph is to provide guidelines as to how pesticide- related disease may be prevented by the monitoring of workers exposed to pesticides. Leslie London, University of Cape Town 5
  • 6. 2. What type of monitoring is available? Broadly speaking, there are two main methods of monitoring exposures to potentially hazardous chemicals a) Environmental monitoring of the chemical or its residue in the environment (air, foliage, soil) or in contact with humans (overalls, skin contact) b) Monitoring of the intact chemical or its metabolite in the tissue or fluids of the body (Biological Monitoring), or the effect of the chemical on enzyme systems within the human body (Biological Effect monitoring) Because there are many variables that determine whether a chemical will be absorbed from the skin or through the lungs into the human body (use of protective equipment, safety practices climatic conditions, individual susceptibility, concomitant disease, properties of the formulation and the chemical. etc), biological monitoring is regarded as a more accurate assessment of human exposure. Once the intact pesticide is absorbed, it may be metabolised, redistributed in the body and differentially deposited within body tissues. For this reason the measurement of the biological effect of a pesticide within the body is regarded as a better indicator of exposure to a pesticide. The sequence of steps in chemical toxicity is illustrated in the figure on the next page. From Leslie London University of Cape Town Leslie London, University of Cape Town 6
  • 7. Figure: Chemical Toxicity in Humans EXPOSURE TO CHEMICAL DEPOSITED ON SKIN OR IN LUNG ABSORPTION METABOLISM REDISTRIBUTED DEPOSITED IN TISSUES EFFECTIVE EFFECTIVE EFFECTIVE TISSUE DOSE ₁ TISSUE DOSE ₂ TISSUE DOSE ₃ BIOLOGICAL BIOLOGICAL BIOLOGICAL EFFECT ₁ EFFECT ₂ EFFECT ₃ From Leslie London University of Cape Town DISEASE 3. What chemicals may be monitored? For most organophosphate and carbamate pesticides, a fairly simple measure of biological effect is available. This is the measurement of the enzyme cholinesterase, which is inhibited by the carbamate and organophosphate pesticides. The enzyme cholinesterase may be measured in the blood (either within the red blood cells, or in the blood plasma surrounding the blood cells). Lowered levels of cholinesterase activity indicate exposure to organophosphate or carbamate pesticides. The mechanism by which these pesticides cause adverse effects is by inhibiting the cholinesterase in the nervous system. By measuring the cholinesterase in the blood, one can determine the activity of the chemical in the body before it has an effect on the nervous system. Leslie London, University of Cape Town 7
  • 8. For most other pesticides, such simple methods of biological effect monitoring are generally not available. lf one wishes to monitor workers for exposure to pesticides other than organophosphates and carbamates, the tests required are complex, time-consuming and costly (involving measurement of the intact pesticide or its metabolite in the blood or urine of the exposed worker, or the intact pesticide in the environment). These tests are generally not widely available in South Africa at commercially practicable costs despite their requirement in terms of the HCS regulations. This monograph concentrates on biological monitoring of exposure to organophosphate and carbamate pesticides. However, it is recommended that where other highly toxic agents are in use (e.g. pentachlorophernol) every attempt should be made to establish a monitoring programme for these agents. 4. Biological monitoring of exposure to organophosphate and carbamate pesticides Organophosphates and carbamates bind to the enzyme cholinesterase in the human nervous system, causing an accumulation of chemical neurotransmitters. This, in turn, leads to an overactivity within the nervous system manifesting as the symptoms of acute poisoning. By measuring the effect of these pesticides on similar enzymes in the blood, one can detect a decrease in enzyme activity before symptoms develop and therefore prevent poisoning Of the two blood cholinesterases, plasma cholinesterase is regarded as most sensitive to recent absorption of Ops and carbamates, while the red blood cell cholinesterase reflects more closely the concurrent effect in the nervous system. Thus both enzymes may be used to prevent nervous system poisoning, but different action levels apply to the two enzymes. This is dealt with in sections 7, 8 and 9. If a monitoring programme is to be used it is preferable that both enzymes are monitored. If financial resources are limited, the red cell cholinesterase is preferred as the best assay because it reflects more closely the physiological effect of the chemicals on the worker’s nervous system. Leslie London, University of Cape Town 8
  • 9. 5. How may one know whether a cholinesterase level is low, or has dropped? Because cholinesterase levels are known to differ widely between individuals, irrespective of their exposure to chemicals, the optimal method of determining what is normal for a person is to establish the individual’s baseline level. This can be compared to enzyme levels after exposure to organophosphates or carbamates in order to determine whether a drop has occurred. The figure below illustrates a hypothetical case where serial testing for cholinesterase level is performed before, during and after exposure. By withdrawing the worker from exposure timeously, he or she may be prevented from becoming symptomatic, and may safely be returned to work once their cholinesterase has returned to normal. Figure: Serial testing for cholinesterase before, during and after exposure organophosphates or carbamates From Leslie London University of Cape Town In the absence of a baseline for an individual worker, it is difficult to determine whether a workers cholinesterase has decreased. Laboratories often quote normal ranges for cholinesterase levels, but because the variation between individuals is so high, these 'normal' ranges are not helpful for biological monitoring. For example, a few workers who have no significant exposures of organophosphates or carbamates may have cholinesterase levels well below the 'normal' ranges simple because of genetic variability. The normal ranges developed for laboratory use are generally based on statistical distributions, Leslie London, University of Cape Town 9
  • 10. and bear little relationship to the practical distributions, and bear little relationship to the practical applications of biological monitoring. For this reason, it is advisable to develop individual baselines for each worker in the monitoring programme, rather than rely on normal ranges to identity exposed workers. 6. How does one establish e baseline cholinesterase level? To be sure that it is a baseline, the person should preferably be tested before starting work. Alternatively, if already in employ, he or she could be tested in the course of the year as long as they have had no exposures to organophosphates or carbamates within the previous 2 or 3 months. This is the time taken for the red blood cell cholinesterase to recover alter significant exposure. It is therefore important that the worker is not tested tor a baseline while his/her cholinesterase is still below their true normal. Besides the natural differences in cholinesterase levels between people, there are also biological fluctuations from one day to another. While these differences are small, it may still affect the level at which you establish the individual’s baseline level, especially if the precision of the method used to measure cholinesterase is low. For this reason, it is better to take the mean of two measurements to establish a baseline level of cholinesterase for subsequent comparisons. 7. How much of a decrease in cholinesterase is significant? Researchers have correlated the level of decline in cholinesterase with the development of symptoms in a series of studies. It is generally agreed that a decline at 40% or more in plasma cholinesterase (i.e. down to 60% of baseline levels or lower) is associated with symptoms, while for red blood cell cholinesterase the decline required to produce symptoms is 30% or more (i.e. down to 70% of baseline levels or lower). [Note the HCS regulations cite a drop to 70% of baseline levels of red cell cholinesterase as the Biological Exposure Index for exposed workers.] In order to prevent the onset of symptomatic disease, declines of less than those described above should warrant preventive actions, which may include investigating the work environment, re-testing the worker or removing the workers from any further exposure. However, because individuals' cholinesterase levels differ slightly from day to day, it is important to be able to distinguish a benign daily fluctuation from a significant decline warranting further action. Research has shown that biological fluctuations should not exceed 10 to 15% from day to day. For this reason, declines of between 10 and 30% (red cell cholinesterase) and between 10 and 50% (plasma cholinesterase) are significant declines that could be used to prompt preventive action. Leslie London, University of Cape Town 10
  • 11. 8. What type of preventive action should be prompted by a drop in cholinesterase? The table below is based on regulations of the California Health Department and lists the preventive actions that should be taken at different levels of cholinesterase decline from baseline. If a worker has been removed from exposure as a result of a decline in cholinesterase, he or she should not return to work until the red blood cell cholinesterase has risen to at least 90% of the baseline value (i.e. within normal variability). It is important to have alternative work available for these workers while their cholinesterase levels are depressed. When a worker’s baseline cholinesterase is based on two readings, its precision is increased and it will be easier to tell whether a small decline in cholinesterase is significant or not. Leslie London, University of Cape Town 11
  • 12. 9. How frequently should testing be done? Once a baseline is established for a worker, it is advisable that he or she be regularly tested during ongoing exposure. Testing should happen at least once during the spray season, preferably around peak spraying time. However, it is advisable to test more regularly than this. For example, California regulations prescribe that workers having more than 6 full days exposure to organophosphates or carbamates within a 30 day cycle, should be tested, or that a worker with more than 40 hours contact with pesticides in a weeks' schedule should be tested. Other factors particular to the work setting may also be used to prompt testing, such as continuous work with pesticides in a closed environment, or known accidental exposure. Note that the HCS regulations leave the choice of the timing and frequency of testing to the discretion of the occupational health practitioner. 10. What technology is available for testing in the field? Usually, cholinesterase testing is available from commercial and University laboratories at competitive rates. This requires the presence of professional staff available to draw blood under ice to the relevant laboratory, and awaiting the laboratory result. One drawback is the potential delay between the taking of the blood and receipt of the result, or, worse still, loss of the sample of the result in transit. An additional consideration is the need to ensure that the laboratories practice adequate quality control of cholinesterase estimation. This is particularly the case for the red cell cholinesterase assay, for which the methodology is fairly complex and susceptible to many sources of error. For this reason, it would be advisable to ensure that the laboratory to which venous blood samples are sent, can give reasonable account of their efforts to ensure quality control, or, preferably, demonstrate that they are part of laboratory quality control programme for the tests in question. Such a programme has been suggested by the National Centre for Occupational Health in Johannesburg. An alternative technology for cholinesterase estimation involved field kits based on finger prick devices. A number of such devices are available, although their reliability and validity are not widely described. One such field kit (the TestMAte OP) has been tested under field and non-field conditions in the Western Cape and shown to have sufficient reliability as to be able to apply the California regulations with reasonable robustness. Such technology may be easily applied by field staff (not necessarily health professionals) with sufficient training, and the benefits of immediate results in the field may outweigh the slight loss of precision involved. Leslie London, University of Cape Town 12
  • 13. 11. How should a monitoring programme be managed? Clearly, the decisions as to who will be responsible for the planning, implementation and evaluation of the programme will lie within the ambit of the workplace organisation. However, certain questions will need to be decided by those responsible for the programme. These would include: a. What workers will be included in the monitoring programme? This will be informed by your risk assessment, required of employers in terms of the HCS Regulations. It is advisable to include all workers who handle pesticides, whether they are involved in the mixing, application, storage or transport of pesticides in a monitoring programme. Other specific indications may be added depending on specific conditions - e.g: Workers who perform maintenance work on spray equipment, or who live in dwellings exposed to spray, etc. Risk assessments to identify the presence of exposures and the workers at risk should be conducted at least every two years, but preferably more frequently. b. How often should testing be done? As indicated above, a baseline should be performed followed by periodic testing, at least once per season, but preferably more often, timed in relation to intense spraying activities. c. What thresholds should prompt preventive actions? Thresholds that should prompt preventive actions are summarised in the table above. d. What follow up is needed? Affected workers must be followed up until their cholinesterases have returned to normal. Investigation of the work place must be done to identify behavioural, structural and organisational sources of exposure amenable to intervention. Appropriate training and hygiene measures should then follow. e. How long should records be kept? Record keeping is an essential component of good industrial hygiene practice as well as comprising an important requirement of the imminent Hazardous Chemical Substance Regulations under the Occupational Health and Safety Act. In terms of these regulations, all workplaces where organophosphate chemicals are used will be required to perform appropriate medical monitoring of exposed workers, and to be able to demonstrate evidence of such monitoring to the department of Labour inspectorate. The regulations insist on records being kept for at least 30 years. Leslie London, University of Cape Town 13
  • 14. Additional reasons for maintaining adequate records are the need for proper documentation for potential compensation requirements, and the potential for many of the cholinesterase compounds to cause long-term chronic health effects. Good monitoring data during the course of employment will be essential in interpreting chronic neurological and other disorders arising in workers, as well as in Assisting future research into many of the long-term hazards of ongoing chemical exposures. f. Informed consent It is important to remember that a monitoring programme involves invasive procedures (the taking of blood) and therefore workers should only participate after giving full informed consent. This is an additional argument for having monitoring programmes supplemented by education and training as part of a comprehensive health and safety programme. 12. What are the common pitfalls in implementing biological monitoring programmes for organophosphate and carbamate pesticides? Biological monitoring will provide a useful means preventing disease due to exposure to organophosphate and carbamates. However, it cannot serve as substitute for other methods of pesticide safety, such as engineering controls, correct use of personal protective equipment, administrative controls and worker education and training, all of which should be integrated in a comprehensive health and safety progamme at the workplace, and which are feasible in the agricultural setting. Once a monitoring system is in place, it should be regularly evaluated to ensure that it is effective and efficient. Too often, programmes run routinely to meet statutory requirements without attention to whether they are achieving what they set out to achieve. Quality control is often neglected in many monitoring programmes, particularly with regard to laboratory results. An efficient programme can be a waste of time and resources if data from the laboratories is invalid. You should insist on evidence of adequate quality control from the laboratories where you have conducting your analyses. If necessary, submit one or more duplicate samples from the same subject with an alias/aliases. By comparing the results, you will be able to get e good quantitative idea of the laboratory’s precision. 13. How should monitoring programmes be evaluated? Depending on the scope of the programme, its objectives and the amount of resources available, evaluation of a monitoring programme could examine different aspects. Leslie London, University of Cape Town 14
  • 15. Firstly, the programme needs to establish that it is achieving the prevention of disease due to organophosphates and carbamate pesticides. Surveillance foe cases of poisonings reported at work and outside the workplace involving exposed workers can provide this type of information. This is a measurement of the effectiveness of the programme. Collaboration between farm health personnel and information flow in order to achieve this objective. Secondly, the process outputs of the programme should be evaluated. How many workers were monitored, how many tests were done and how many preventive actions (retesting, workplace investigations, withdrawal from exposure, etc) were triggered by the programme? This is an indication of how many workers were protected from potential disease by the application of the programme. Thirdly, how efficient was the programme implemented? Were tests performed in the correct manner, and at the appropriate times? Were the correct workers monitored? Were there any important omissions from the programme? Fourthly, what were the costs, direct and indirect, and what were the benefits in financial terms? Lastly, what was the functional benefit to workplace morale, productivity, occupational health service staff job satisfaction etc? 14. A checklist before implementing a monitoring programme Based on the above, it is helpful to use a checklist of questions before implementing a monitoring programme:  What are the objectives of the programme? Are they clear?  lf prevention is to be the objective of the programme, what protocols will be followed to ensure that the objective can be met? Are these protocols clear to all personnel involved?  Will the information recorded enable you to audit the programme?  Is the programme linked to an education package?  Is the programme going to be sustainable? 15. Diagnostic Tests compared to Biological Monitoring There are occasions when testing will be used to detect the presence of illness in workers exposed to organophosphate and carbamate pesticides. For example, workers presenting to the farm nurse with non-specific symptoms such as headache, nausea or dizziness may be suffering from early or mild organophosphate poisoning. Under such circumstances, cholinesterase testing may be used to diagnose the baseline level is available, comparison of the result will enable the clinician to make a simple diagnosis. Leslie London, University of Cape Town 15
  • 16. However, clinicians are often faced with a situation where no baseline is available, and where the single cholinesterase result is within or only slightly below ‘normal’ laboratory range. Under these circumstances, serial testing of cholinesterase after withdrawing the worker from exposure may be useful to confirm mild cases of poisoning. If the cholinesterase level gradually increases by more than 15 % over time, and then plateaus out with a retrospective confirmation of cholinesterase inhibition as the cause of the worker's illness. This is clearly of no use in trying to prevent poisoning (the purpose of biological monitoring), but is useful in the clinical setting of trying to reach a diagnosis. Note that all cases of occupational poisoning including those diagnosed retrospectively must be reported to the Chief Inspector: Occupational Health and Safety, Department of Labour in terms of the OHSA. At the present time, remember also that any cases of pesticide poisoning are notifiable to the Department of Health in terms of the Health Act. From Andrea Rother University of Cape Town Leslie London, University of Cape Town 16
  • 17. SUMMARY PROTOCOL FOR A BIOLOGICAL MONITORING PROGRAMME FOR WORKERS EXPOSED TO ORGANOPHOSPHATE CARBAMATE PESTICIDES Planning Monitoring Programme: Objectives Who participates Frequency of testing Alternative job sites Responsibilities for investigation Training How to evaluate What technologies to use Baseline Cholinesterase Testing: Pre-employment or at least 2 months without exposure Interval testing: Every month it more than 6 days exposure Per month Other indications Depending on level of Cholinesterase: Re-test Investigate Withdraw from exposure Wait till CHE back to normal Leslie London, University of Cape Town 17
  • 18. Bibliography Ames RG, Brown SK, Mengle DC, Kahn E, Stratton JW, Jackson RJ. (1989). Protecting agricultural applicators from overexposure to cholinesterase-inhibiting pesticides: Perspectives from the California Programme J.Soc Occup Med. 39, 85-92. Ames RG, Brown Sk, Mengle DC, Kahn E. (1989b) Cholinesterase activity depression among California agricultural pesticide applicators. Am J Ind Med. 15 ; 143-l50. Brown SK, Ames RG, Mengle DC. (1989). Occupational Illness from Colinesterase-inhibiting pesticides among agricultural applicators in California, 1982-1985. Arch Env Health, 44: 34 - 39. Coye MJ, Lowe JA, Maddy KT. (1986a). Biological Monitoring of agricultural workers exposed to pesticides: 11. Monitoring of intact pesticides and their metabolites. J Occup Med 28(8): 628 - 442. Fillimore Cm, Lasenger JE. (1993). A cholinesterase testing programme for pesticides applicators. J Occup Med. 35: 61 -70. Hulka BS, Margolin Bh. (1992). Methodological Issues in Epidemiological Studies using biologic markers. Int J Epidem 135 : 200- 209. Krieger Ri, Ross JH. (1993). Risk assessments in the pesticide regulatory process. Ann Occup Hyg. 37: 565 - 578. Lotti, M (1992). Central neurotoxicity and behavioural effects of anticholinesterases. ln: Clinical and experimental toxicology of organophosphate and carbamates. Eds Ballantyne B, Marrs TM, Aldridge Wn. Butterworth, Stoneham, MA. Minton NA, Murray VSG (1988). A review of organophosphate poisoning. Medical Toxicology 3 : 350 - 375. Toxicology. Special edition as a Manual on Biological monitoring for Pesticides. 17 June 1994. 19(1). Woolen Bh. (1993) Biological monitoring for pesticide exposure. Ann Occup Hyg. 17 : 525 - 540. World Health Organisation (1986b). Organophosphorus insecticides : A general introduction Environmental Health criteria 63. World Health Organisation, Geneva. We would appreciate your feedback for this Open Educational Resource (OER), by completing this form. Alternatively, you can email us at healthoer@uct.ac.za Leslie London, University of Cape Town 18