EPA-650/1-74-009
June 1974
Environmental  Health Effects Research Series


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                                     EPA-650/1-74-009
PESTICIDES  EPIDEMIOLOGICAL
            FIELD  STUDIES
                     by

               Dr. John E.  Davies

        University of Miami School of Medicine
              1600 N. W. 10th Avenue
               Miami, Florida  33152


                Grant No.  802112

                     and

              Contract No. 68-0211145
                ROAP No. 21AYL
            Program Element No. 1E1078


      EPA Project Officer: Dr. William F. Durham

    Pesticides and Toxic Substances Effects Laboratory
        National Environmental Research Center
     Research Triangle Park,  North Carolina  27711


                 Prepared for

       OFFICE OF RESEARCH AND DEVELOPMENT
     U.S. ENVIRONMENTAL PROTECTION AGENCY
            WASHINGTON, D.C. 20460

                   June 1974

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This report has been reviewed by the Environmental Protection Agency
and approved for publication.  Approval does not signify that the
contents necessarily reflect the views and policies of the Agency,
nor does mention of trade names or commercial products constitute
endorsement or recommendation for use.
                                  ii

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                                   Abstract

 Quantisation of  pesticide exposure Is  an essential  feature  of  occupational  health
 Insofar as the safety of  the pesticide worker  is  concerned.  When  accurate  pesti-
 cide exposure can be obtained surveillance will protect  the pesticide  applicator
 and  has the potential for evaluating human pesticide  residue exposures and  the
 reliability of current re-entry  standards.   Since worker exposures are usually
 multiple,  the use of the.raultiresidue  technique for the  analyses of urinary meta-
 bolites can provide  the key  to quantisation  of mixed  organophosphate and  carbamate
 human exposures.   With this  philosophy in mind, occupational studies of pilots
 and  loaders were measured.   Aircraft loaders were found  to  be  receiving excessive
 pesticide  exposure as evidenced  by the occurrence of  several acute pesticide intoxi-
 cations,   a chronic  state of cholinesterase  inhibition and  a high  turnover  in this
 category of work.  The study of  urinary di-alkyl  phosphates  and phenols confirmed
 the  excessive exposure of these  two working  groups  and the  amounts of  di-ethyl
 phosphate  (DEP),  di-ethyl thiophosphate (DETP), dl-methyl phosphate (DMP) and
 di-methyl  thiophosphate (DMTP) and paranltrophenol  (PNP)  were  significantly in-
 versely correlated with the  red  blood  cell cholinesterase at the p 0.001  level.
 Thus,  the  lower  the  red blood  cell cholinesterase,  the higher  the  urinary meta-
 bolites.

 The  four di-alkyl  phosphates were  the  derivatives constantly identified in  these
 occupational exposures.   Following an  eight  hour  exposure to a single  organophos-
 phate both the di-alkyl phosphates and the appropriate phenolic derivatives per-
 sisted  in  the urine  for thirty six hours  or  longer.  The  impression was gained that
 the  optimum time  for urinary surveillance based on  a single  sample was  nine hours
 after the  last exposure.  These  two groups of metabolites were also exceedingly
 informative  not only in the  confirmation  of  acute pesticide  poisoning but also
 in the monitoring  of the  poisoned  victim with regard to his  need for antl-dotal
 therapy.

 Organophosphate poisoning is usually thought of as  being  a abbreviated episode
 wherein the  clinical manifestations and outcome are usually  resolved within  48-
 72 hours.  Recently  we  have  encountered a much more protracted illness following
 acute exposure to  certain halogenated  organophosphates, this was especially
 noticeable following intoxication  of VC-13W which is a dichlorophenol di-
 ethyl phosphorothioate.  The presence  of the halogen derivatives attached to
 the phosphate moiety resulted in the delayed occurrence of acute intoxication
 and a protracted clinical crisis.  This was  the case in the poisoning reported
 herein, and emphasizes  the need for a  changed appraisal of human subjects in-
 toxication by  these  fat soluble organophosphates.   Contrary  to past experience
 VC-13'*1) and Dursban^ were regularly detected in the air sample and DDT was
 conspicuously absent.  Qualitatively the air samplings reinforced  the greater
 use of organophosphates in South Florida.
This report was submitted in fulfillment of Project Number 68X0107 Grant Number
R802112 and of Project Number AS0871 Contract Number 68-02-1145, by the Univer-
sity of Miami School of Medicine, under the sponsorship of the U.S. Environmental
Protection Agency.  Work was completed as of December, 1973.
                                       ii

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                          CONTENTS








                                                      Page




Abstract                                              11




List of Figures                                       Iv



List of Tables                                         v



Acknowledgements                                      vl



Sections



I     Conclusions                                      1



II    Recommendations                                  3



III   Introduction                                     4



IV    Urinary Metabolite Studies                       5



V     Air Sampling Studies                            27



VI    EEC Studies                                     32



VII   Acute Pesticide Poisoning Investigations        34



VIII  References                                      40



IX    Glossary                                        41
                             iii

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                               FIGURES


No.                                                             Page

 1     Monthly red blood cell and plasma cholinesterase,
       paranitrophenol and alkyl phosphate metabolites
       observed In a helicopter crop duster (F.B.),
       Belle Glade, Florida, 1973                                12

 2     Comparisons of cholinesterase and paranitrophenol
       excretions observed in an aircraft loader (F.R.),
       Belle Glade, Florida, 1973

 3     Comparisons of cholinesterase and paranitrophenol
       excretions observed in an aircraft loader (K.D.T.)
       Belle- Glade, Florida, 1973                               14

 4     Combined alkyl phosphate excretion in an aircraft
       loader (M.W.), during ethyl-methyl parathion exposure     16

 5     Urinary excretion of Iso-propoxyphenol (Baygon 0.5%)
       in a structural pest control operator (Max P.)
       following 6 hours of exposure                             21

 6     Urinary excretion of Iso-propoxyphenol (Baygon 0.5%)
       in a structural pest control operator (Edward S.)
       following 6 hours of exposure                             22

 7     Comparisons of urinary 3,5,6-TC pyridinol metabolite
       and alkyl phosphate metabolites observed in a structural
       pest control operator (M.P.), Miami, Florida, 1973        24

 8     Comparisons of urinary 3,5,6-TC pyridinol metabolite
       and alkyl phosphate metabolite observed in a structural
       pest control operator (E.S.), Miami, Florida 1973         25
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                                  TABLES
No.                                                                     Page

 1     Number of Urinary Metabolite Analyses and Blood Cholineaterase
       Studies in Different Pesticide Exposed Groups in South Florida   g
 2     Comparisons of Urinary Alkylphosphate Concentrations in
       Intermediate Exposed Workers and Controls in South Florida      LQ

 3     Correlation Coefficient (R) of Paranitrophenol, Red Blood Cell,
       Plasma Cholinesterase With Urinary Alky 1 phosphates From Pesticide
       Exposed Workers                                                 H

 4     Cholinesterase and Urinary Isopropoxyphenol Levels in Baygon
       (0.5%) in Structural Pest Control Operators and Assistants      ^9

 5     Twenty-four Hour Excretion of Isopropoxyphenol in Two Spraymen
       Occupational ly Exposed for  Six  Hours to 0.5% Baygon Spray
       Solution                                                        20

 6     Thirty- two Hour Urinary Excretions of 3,5,6-TC Pyridinol and
       DMP, DEP, DETP in Two Structural Pest Control Operators         23

 7     Retention of Pesticides in Ethylene Glycol (12 hr. run=Flow .82
       CFM (16.4 m3)                                                   29

 8     Ethylene Glycol Trapping Efficiency (12 hr. run=Flow .82 CFM
       (16.4 m3 of air)                                                30
                                             3
 9     Air Concentrations of Pesticides (ng/m ) from Different Sampling
       Sites in South Florida 1973                                     31

10     Confirmed Fatal Intoxications in South Florida, December 1972  -
       December 31, 1973                                               35

11     Confirmed Non-Fatal Intoxication in South Florida, 1973         37

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                            ACKNOWLEDGEMENTS
The investigators acknowledge with gratitude the cooperation and assistance
offered us to conduct these studies by Allied Helicopter Service Inc., the
Dade County House and Urban Development Program, the Eveready Pest Control
Company.  Without the enthusiastic support of the management and staff of
these institutions, these evaluations of human pesticide exposure under
normal working conditions could not have been possible.
                                  vi

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                               SECTION  I

                               CONCLUSIONS

 Important  conclusions were  reached  In three of  the four areas of the  research
 conducted  this  last year.   In  the area  of  the study of urinary alkyl  phosphates
 and  phenolic derivatives, there Is  no doubt that recent multi-residue analyti-
 cal  techniques  have provided a greater  opportunity to measure human occupational
 exposure to pesticides.  The data facilitates the quantltatlon of mixed pesti-
 cide exposures  from workers involved in pesticide application, and from workers
 exposed to dislodgable  leaf residues in re-entry situations.  Their measurement
 also significantly contributed to the accurate  diagnosis of acute pesticide
 poisoning  and in  the Investigation  of suspect "pesticide associated"  illnesses.
 They will  be useful to  the  physician as well, for their presence is informative
 during the clinical management of the poisoned  victim; they provide an indica-
 tion for the continued  need for atropine and oxime therapy.

 In the area of  occupational pesticide exposures, our preliminary data although
 based upon small  numbers suggested  that the alkyl phosphates are significantly
 inversely  correlated with red  blood cell cholinesterase levels.  When the pesti-
 cide exposures  of the aircraft loaders  were compared with the pilots, the ex-
 cessive exposure  of the former was  substantiated by a) the number of  acute poi-
 soning episodes,  b) an  88%  employee turnover rate, c) very low cholinesterase
 levels particularly the red blood cell  values,  d) high paranitrophenol averages,
 and  e) by  DEP,  DETP and DHP alkyl phosphate residues.  The association of DMP
 with red blood  cell cholinesterase  values was particularly impressive and was
 an association  which possibly  emphasized the special toxicity problems of
 Phosdrin(R)>  As  was the first experience of Dr. Shaflk and his colleagues,
 in animal  studies, OEP, DETP,  DMP and DMTP were the only alkyl phosphates found
 under these conditions  of mixed organophosphate exposures.  The monoethyl and
 monomethyl phosphoric and phosphorothioic acids are not discussed herein because
 Dr.  Shaflk stated that  they are found in acute  pesticide poisonings only and
 even then  their sensitivity is poor, and therefore, there is no need  to look
 for  other  alkyl phosphates  other than those herein described.  Another signi-
 ficant finding  was the  possible significance of DETP/DEP ratio; greater amounts
 of the former were observed in workers when their exposure was Co ethyl and methyj
 parathion.  At  this point in time,  this observation must be considered as an im-
 pression rather than a  conclusion and it certainly warrants further study.

 Since exposure  to parathion was the predominant organophosphate exposure in
 the  applicator  group this exposure  promoted an  evaluation of the relative
 merits of  the alkyl phosphate  data  compared to  the paranitrophenol levels.
 Since grab samples of urine rather  than 24 hour urine outputs were studied,
 firm conclusions  cannot be  reached  on the relative merits of either type of
 metabolite.  During the dally  studies of alkyl phosphate and paranitrophenol
 levels in  the urine of  one  loader,  the Impression was gained that the former
 particularly DEP  plus DETP  were less subject to the wide variations observed
 with paranitrophenol values.   One conclusion which can be made with some de-
 gree of certainty is that more field research is necessary so that diurnal
 variations of both types of metabolites can be studied especially following
work exposure to a single pesticide; a parathion urinary profile is the one
most urgently needed.   Another important conclusion was that these metabolites
promote the study of organophosphate and carbamate exposures at levels not
necessarily producing cholinesterase inhibition.  With such features,  there-
 fore, they are ideal for occupational surveillance and for future use as
epidemlologic Instruments.

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The special studies of Baygon and Dursban made it possible to conclude that
future surveillance programs with these pesticides, if reliance must be placed
on a single grab sample of urines, then 6 to 9 hours after exposure is the
ideal time to collect such a specimen.  With regard to urinary pesticide
metabolites concentrations should be expressed in terms of micrograms per
hour rather than by correcting for urine osmolality.  Pesticide residues were
much greater than expressed in this way then when corrected for an osmolar
correction factor.

From the epidemiological studies of acute pesticide poisoning, several con-
clusions were reached.  The number of confirmed cases and the growing problem
of   pesticide suspected Illness emphasize the future need for Pesticide
Poison Control Centers in those areas were pesticides are heavily used.
New pesticides are appearing on the scene and without a system for monitor-
Ing the acute incident it is Impossible to anticipate generalized effects in
an area before they occur.   VC-13 is a case in example—the different clinical
consequences of over exposure to this fat soluble pesticide was first appreciated
as a result of the Investigation of an acute poisoning.  The more halogenated
ions are in the pesticide moiety the more delayed are the effects.  Our exper-
ience with VC-13 poisoning confirms that chronic organophosphate pesticide
poisoning has now become a reality.

New pesticides are now found in the South Florida ambient air such as VC-13,
Dursban and some of those identified are fat soluble and the whole picture
is changing from an incidental organochlorlne potential to an incidental
organophosphate exposure.   One must conclude that an ongoing meaningful air
sampling program is essential.

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                               SECTION II

                             RECOMMENDATIONS


The potential of the urinary pesticide metabolite needs to be further ex-
plored In the occupational environment of the pesticide applicator, in the
work situation following exposure to dlslodgable residues and as an evalua-
tory parameter of proposed field re-entry studies.  In the presence of
these needs further studies on urinary metabolite excretion patterns under
normal working conditions should be conducted.  Special emphasis should be
given to single pesticide exposure situations - 36 to 48 hour urinary pro-
files.  Methodologic research of urine metabolites In the direction of a
simplified and accelerated method of testing is urgently needed.  These
should be applicable for testing in the field, what is really needed is
a sort of dipstick test for occupational exposure.  Until this is developed
grab sample urines, a system which requires knowledge of the prior voiding
time,seemsto offer the only acceptable type of specimen in a large scale
surveillance program.  Based on the findings of this first year's work
the metabolites must be evaluated for as long as 48 hours after a six hour
exposure.  We have no Information on the different rates of excretion of
alkyl phosphates which are the result of differences of fat solubility.
We do not know whether there are diurnal variations in the concentrations
that are excreted following a single exposure nor have we confirmed that
these metabolites are present in the general population.  It is recommended
that these studies be directed to answer some of the aforementioned ques-
tions  to be conducted as soon as possible if the merits of the urinary meta-
bolite is to be measured in human exposure studies.

In addition to the development of metabolite information, our emphasize
this year has been the need to establish selected pesticide poison con-
trol centers in appropriate areas where pesticides are extensively used
In the United States.  Secondly, with the change of pesticide use patterns
an ongoing air monitoring program should be instituted in the areas of
high pesticide usage in the United States.

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                               SECTION III

                               INTRODUCTION
SCOPE AND PURPOSE OF THE PROJECT

This project entitled "Pesticide Field Epidemiologlc Unit" has as its primary
objective to Implement field epidemiologic studies of in-house research areas
recently developed in Che U.S. Environmental Protection Pesticides Laboratory
in Perrine, Florida.  The subordinate objectives reflective of in-house research
areas which were ready for field testing included:

      A.  Urinary metabolite studies of pesticide workers
      B.  Air Monitoring studies
      C.  Field testing of a portable EEC apparatus
      0.  Pesticide poisoning investigations.

This was a new three year project, and in the first year called for the reloca-
tion and establishment of a new pesticide laboratory in the University of Miami
School of Medicine.  This annual which covers pertinent findings in the first
year in these four areas of pesticide field epidemiologic investigations must
aarve as a final report since owing to budgetary cuts in projects, the second
and third years of the study were renegotiated on a contractural basis.

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                                SECTION IV

                       URINARY METABOLITE STUDIES
 A.   Urinary metabolite studies - Background  Information.   Traditionally,  the
 occupational health surveillance of  the occupationally  exposed  worker has
 relied  upon the red cell  and  plasma  cholinesterase changes observed under
 conditions  of pesticide exposure as  an indication of  human exposure to organo-
 phosphate and carhamate insecticides.   Experience has shown that  there are
 several serious limitations of the interpretability of  these indices.  These
 included the wide range of normal cholinesterase  values in the  non-exposed
 individual, the absence of a  definitive cholinesterase  level which  is diag-
 nostic   of  the occurrence of  overt illness,  and the necessity of  obtaining
 pre-exposure baseline  data in occupationally exposed  studies.   Because of
 this, many  workers In  occupational health have investigated the use of
 pesticide urinary metabolites as an  alternate human pesticide exposure in-
 dex.  Parathion,  carbaryl and propoxur are examples of  some of  the  more
 commonly used pesticides  in this group whose metabolites have been  studied
 under conditions of occupational exposure. Recently Shaflk,  Enos  and their
 colleagues  have developed a residue  procedure for halogen  and nitrophenols
 in urine.       Their preliminary data  suggested that  these newer  techniques
 lend themselves to a more specific study of  pesticide exposure  in man,  and
 may well prove to be the  key  to the  quantltation  of mixed  organophosphates
 and carhamate exposures.

 In a preliminary study  of six samples  from persons having  no exposure and
 from  occupational workers exposed to  Dasanlt (Fensulfrothlon), Thimet
 (Phorate) and Di-Syston (Dlsulfoton),  Shafik found that DMP,  DEP, DETP
 and DMTP alkyl phosphate metabolites were identified.   In  the non-exposed
 group urinary metabolite  concentrations ranged from 0.005  - 0.04, from 0.003 -
 0.08, from  0.02 - 0.10  and from Not Detected - 0.01 ppm respectively for
 these four  alkyl  phosphate metabolites.  The average  concentrations for these
 metabolites in this non-exposed group  was 0.01, 0.05, 0.06 ppm  and  Not  de-
 tected  respectively.  Similarly from six samples  of the occupationally  ex-
 posed,  the  ranges were  0.005-0.06, 0.34-2.41, 0.06-0.22,  and 0.16  and
 1.62 ppm respectively.  Average concentrations in these six samples from
 the exposed were  0.02,  0.97,  0.10, and 0.60  ppm respectively.   These very
 preliminary data  suggested that these  metabolites  reflected  occupational
 exposure  to organophosphate pesticides and were now ready  for field testing.

 Our studies have  sought to take over from this point  and in  a logical step-
 wise progression  to explore in  greater detail the  qualitative and quantita-
 tive profile  of  these pesticide metabolites  in several  occupational groups
 in South  Florida.   In addition, we have  attempted  in  a  preliminary  way  to
 explore  the relationship of these metabolites to  cholinesterase findings.

Description of Phases - As soon as we received the grant award,  we  started
 to prepare  our study as follows:

    Phase I - Making contact  with the several companies  in this  area and
              investigating what is their season,  types  of pesticides that
              are being used  and whether they would participate  in our
              study.  It also required the specialized training  of our

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               chemists with Drs.  Shafik and  Enos  in the  multiple  residue
               and alkyl phosphate analytical procedures.   Flans were also
               made to equip and  relocate the pesticide laboratory to the
               University of Miami Medical Research  Building.

     Phase II - Related to the qualitative and some  quantitative examina-
               tions of urines from occupationally exposed  pesticide  workers.

     Phase III - Called for the relation of these  exposure  indices to
               cholinesterase findings.

     Phase IV - Called for individual  24 to 36 hour  urinary pesticide meta-
               bolite profiles for each  individual pesticide.

     Phase V  - Called for the identification  of the  optimal time between
               and after exposure  for  the meaningful ongoing  surveillance
               of the pesticide worker.

     Phase VI - The final phase called for the identification  of urinary
               concentrations of Individual pesticide metabolites  which
               were premonitory of cholinesterase  decline and  the  occurrence
               of acute symptoms.   In  order to systematically  Implement
               the above experimental  design,  the  following studies were
               conducted In a group of pesticide workers in a  helicopter
               spray outfit in  the Belle Glade area  and In  selected ex-
               posure  studies In structural pest control operators.

Materials and  Methods

The  preliminary  qualitative  and quantitative  urine  studies acquired  from
occupational pesticide exposure were  first conducted in cooperation  with
the  Allied Helllcopter,  Inc.,  In  Belle  Glade,  Florida.  This  is a crop
dusting company  which  contracts with  South Bay Growers to  aerially apply
a wide variety of  chemicals  to the  fields  in  the  area.  The individual
growers select and  purchase  the pesticides which  they want to have applied
and  the helicopter  company is merely  responsible  for its application.
The  helicopter company  operates from  a modern helicopter pad  situated
two  miles  outside of Belle Glade  and  they have constructed a new hanger
on the site as well as  adjoining  administrative offices.    Six multiple
rotary blade Bell helicopter aircraft are  used these are fitted with  twin
multiple  Jet spray  beams and twin loading  tanks.   When not in use, the
helicopters are  tethered outside  the hanger.    Inside the hanger there is
a repair  shop and a mechanic is employed full-time.   Showers and changing
rooms with individual  lockers are provided for the four regular pilots
and  their  loaders.  Agricultural chemicals are stored away from the hanger
on a separate site.  During the season the pilots and loaders work seven
days a week.   Two sorties are made daily, the first  at 6  a.m.  until about
10 or 11 a.m. and the second at 2 p.m. to 6 p.m.   The pilots do not mix
the  chemicals themselves and wear coveralls,  a crash helmet,  respirator
mask, gloves and flying boots.  They are partially enclosed in a plastic
bubble and each pilot works with one loader.   The loader  decants the agri-
cultural chemical from 55 gallon drums of the pesticide concentrate in
the  airport into the drum of a large tank truck.   In addition, five gallon
drums of a vide variety of pesticide concentrates are also stored  and trans-
ported on  tne vehicle to the various operational  sites in the field.   The
planes are loaded and reloaded at these sites. Water is  obtained  from ad-

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 joining  canals  and  after being pumped  into  the  tank  the pesticide  concentrate
 is  added and  the mixture agitated.  The diluted material  is  then pumped  through
 a 50  foot hose  into the twin  tanks on  each  side of the helicpter.  These are
 filled and refilled three  or  four times an  hour.  The loaders are  provided
 with  gloves,  masks  and rubber boots.   Additionally,  oceralls were  provided
 during the last six months of the study period.  A wide variety of pesticides
 are applied and the materials used last year  included: parathlon 8-E, para-
 thion 6-3 (ethyl-methyl mixture), parathion-toxaphene mixture, Cygon  (dimetho-
 ate), AAtrex  (atrazine), Trithion (carbophenothion), Phosdrin (mevinphos),
 Dithane,/maneb), Lannate (methomyl), DipelOO,  Azodrin (monocrotophos),
 Monitor W, Dlbrom  (naled), Guthion (azlnophos  methyl), and  2,4-DW.
 After obtaining a signed consent form  from  sixteen employees of the Company
 a study  was conducted on four pilots,  five  loaders and three controls.
 Periodic blood  chollnesterase and urine samples were obtained from this
 occupatlonally  exposed group.  Ten cc. of blood were collected in  a heparinzed
 vacutainer tubes, transported over 100 miles  in an icebox to the University of
 Miami Pesticides Laboratory were they  were  analyzed  24 hours after collection.
 Periodic grab sample urines were collected  in labelled hexane washed jars
 stored in dry ice for up to four days  and then  shipped to Miami where they
 were  frozen at  -15   C and  at  a later date analyzed both for  phenolic deriva-
 tives and alkyl phosphate metabolites.

 The Michel method was used for red blood cell and plasma  chollnesterase  deter-
 minations,  and  the  Shaflk, Bradway, Enos &  Yobs modification for the gas  ,
 liquid chromatography analyses of alkyl phosphate metabolites in urine.  '
 The Shaflk, Sullivan and Enos procdure was  used for the analyses of halo and
 nitrophenols  In urines  and the Shaflk, Sullivan and Enos procedure for  1-
 Napthol  In urine was adapted  to measure Iso-propoxyphenol (IPP). 5  Grab
 sample urines were  corrected  to an osmolality of 800 ml/Osm/1 prior to
 analysis.

 Urine and  blood studies were  conducted in two groups of structural pest con-
 trol  operators  exposed to a single organophosphate or a single carbamate
 insecticide.  Blood  and grab  urine samples were obtained  from six  structural
 pest  control operators (SPCO) working  for the Dade County Housing and Urban
 Development program  who had sprayed tenement  building eight  hours a day with
 a 2%  solution of Dursban and  a 1.5% Baygon  (propoxur); search was made for
 3,5,6-TC pryldinol and IPP during these exposures.  Continuous urinary meta-
 bolite excretion profiles over a 36 hour period were obtained from two SPCOs
 of  the Eveready Pest Control  Company selectively exposed  to  0.5% Baygon, 2%
 Dursban and 0.75% DDVP.

 Results

 Table 1 lists the total number of urinary metabolite analyses and Cholin-
 esterase studies in  the several occupational  groups.   Cholinesterase studies
were not always obtained on every urine specimen collected.   In addition,
 118 chollnesterases were analyzed from pesticide exposure groups including
 agricultural inspectors from  the U.S.  Department of Agriculture, sprayers
 from the Dade County Department of Parks,  etc.  Clinical  surveillance of
 and pesticide metabolite data from the helicopter group of workers revealed
 that pesticide exposure in this group of workers,  particularly the aircraft
 loaders,  was extensive.   Three episdoes of acute pesticide intoxications, all
 in pesticide loaders, occurred in the study period;  34% of 54 red blood  cell
 Cholinesterase determinations in this group gave a delta pH per  hour of  0.40
or less.   A high turnover rate was observed during this study period;  88%
of the loaders and 28% of the pilots  changed  their jobs during a period  of

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                            Table  1.  NUMBER OF URINARY METABOLITE ANALYSES AND BLOOD CHOLINESTERASE
                                      STUDIES  IN DIFFERENT  PESTICIDE EXPOSED GROUPS  IN SOUTH FLORIDA
Analyses
Urinary Phenols
Urinary Alkylphosphates
Blood Chollnesterase
(red blood cell and plasma]
Aircraft Applicators
(n=21)
87
39
54
SPCO
HUD (n=6)
13

11
SPCO
Eveready (n=2)
32
19
6
Others


118
Total
132
58
189
00

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 12 months  and were  lost  to  follow-up.  To  the observer  it was obvious  that  signi-
 ficant  loader exposure occurred during the refilling of the aircraft with pesti-
 cides;  an effect which  was due to  the down draft produced by the rotary wings
 causing an aerosol  exposure from pesticide spillage on  the ground.  This spillage
 could be avoided if an automatic nozzle cutoff valve were a mandatory  requirement
 in all  aircraft spray booms, a suggestion  which was reported to  the Project Officer.
 Sequential data was available on twelve subjects, these were 5 loaders, 4 pilots
 and  3 controls.  The mean and ranges of the red blood cell chollnesterase,  the
 urinary alkyl phosphate  and paranitrophenol concentrations of these subjects
 are  shown  in Table  2.  These subjects during the study  period are typical of
 high, intermediate  and non-organophosphate exposures.   Both the  RBC and urinary
 pesticide  metabolite data emphasized the excessive exposure of the loaders.  The
 four di-alkyl phosphates (DEP, DETP, DMP and DMTP) were the only ones  tested in
 these samples.  Except for  DMTP all were found in each  of the 27 urines analyzed
 in the  loaders; some of  these specimens included pre-exposure 6:00 a.m. voidings.
 Paranitrophenol (PNP) was likewise  found in every specimen in this occupational
 category.  DMTP was not  found in 14 of the 27 urines from the loaders, a de-
 ficiency which probably  reflected early analytical problems of separation of
 the  metabolite and  which was subsequently  cleared up.   In the pilots,  these
 biological indices  predominately of ethyl  and methyl parathion exposure was
 considerably less and on four occasions DMP was not Identified.  DEP was Identi-
 fied on every occasion as was also PNP.  The correlation coefficients  of PNP,
 RBC  and PI. ChE with these  urinary metabolites are shown in Table 3.   Signi-
 ficant  exposure to  organophosphates resulted in the lowering of  the RBC and
 PI.  cholinesterases and  also exposure to parathion as high levels in PNP were
 found.  Thus, If urinary alkyl phosphates  were equally  good measures of occupa-
 tional  exposure to  anti-cholinesterase, one would expect significant inverse
 correlation of them with RBC and PI. ChE and a highly significant correlation
 with PNP.  Conventionally excessive exposure to organophosphates is reflected
 by the  red blood cell and plasma cholinesterase inhibition.  If  the organo-
 phosphate  exposure  is from  parathion, urinary paranitrophenol levels is' an
 additional index of pesticide exposure and high levels  of these  metabolites
 are  found.  This Indeed  was the case and DEP, DETP and  DMP were  significantly
 correlated with these known biological Indices of pesticide exposure at the
 highly  significant  level of p<£ 0.001.  The only exception was in the correlation
 of DMTP data which  almost certainly reflected initial analytical difficulties
 with this  specific metabolite.  The Inverse correlation of red blood cell
 cholinesterase with paranitrophenol on the basis of 15  samples from ten sub-
 jects in this study group was -.76.  Although the occupational exposure to
 this group were to a wide variety of pesticides, ethyl  and methyl parathion
 (6-3 mixture) were  the preparations most frequently used.  The highly  signi-
 ficant  correlations herein  shown strongly  support the potential  of these
metabolites in occupational studies of organophosphate  expsores, although
 the  smallness of sample  size should^be emphasized.  The  highly significant
 inverse correlations of DMP with RBC ChE is especially  noteworthy.  It is
 possible that Phosdrin^  ' exposure which is known to be  highly toxic and
which would give DMP urine metabolites only might be contributory to the signi-
 ficance of this association.

Figure 1 illustrates the monthly levels of urinary metabolites obtained on a
grab sample basis from a pilot who was studied over a three month period of
 excessive spraying.   Monthly red bloqd cell and plasma  cholinesterase  are
 also depicted.  Figures  2 and 3 are first expressed with urinary di-alkyl
 phosphate and phenolic data detected in these helicopter pilots  and swampers
on a grab sample basis.  Corresponding red blood cell and plasma cholinesterases
were not obtained on every occasion that the urine samples were  collected.

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                    Table 2.   COMPARISONS OF URINARY ALKYLPHOSPHATE CONCENTRATIONS IN HIGH,
                              INTERMEDIATE EXPOSED WORKERS AND CONTROLS IN SOUTH FLORIDA.


Exposure
groups
High-
Loaders
(n=5)***
Intermediate-
pilots
(n=4)
Controls
(n=3)
Cholinesterase*
RBC
No. mean,
tests ranee

15


9

3


0.3
:09-.72)

0.7
(.5-. 8)
0.8
(.80-. 82)
Alkylphosphates (ppm)**
DEP DETP DMP DMTP
No. mean, mean, mean, mean,
tests

27


11

3

ranee ranee ranee .range

0.48
(.10-1.37)

0.26
(.07-. 66)
0


0.49
(.11-1.7)

0.11
(.001-. 3)
0


0.25
(.005-. 62]

0.06
(0-.26)
0


0.09
(0-.36)

ND
0
0


Paranitrophenol (ppm)**
No. Mean,
tests ranee

61


11

3


0.8
(.04-4.36)

0.3
(.04-. 74)
0

* pH/hr
**corrected to 800 mOsm/1
***includes 3 acute Illnesses

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           Tfcble 3.  ODREBLAXIflH OOEPFICIEIR (R) OP PAPAT"*OF"™»ff t »F» W-QQn CELL, PLASMA. GBOUH-
                     ESTRASB. HUH URINARY ALKfLPBQSPHAXES FROM PESTICIDE EXPOSED
Biological         	Alkylphosphates  (ppm)	
indices	n    PEP     DBTP   BMP    MtTP   PEP 4- DETP    1BIP + PMTP     PEP. DETP. BHP. BMTP

PHP          39    .75      .75    .5     .54      .80           .66                   .79
(pp.)

RBC ChE
             15   -.72     -.64  -.89    -.11     -.72          -.86                 -.78

Plasma ChE   15   -.52     -.48  -.75     .04     -.53          -.68                 -.58

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   Figure 1.  Monthly red blood cell and plasma cholinesterase, paranitrophenol
            and alkylphosphate metabolites observed In a helicopter crop
            duster (F.B.), Belle Glade, Florida 1973
   PPM
A pH/hour

1.00 r
075
0.50
0.25
         6
        FEB.
                                PLASMA  Che
28

 DATE
   29
MARCH
                                                           j
  30
APRIL
                             12

-------
   Figure 2.  Comparisons of cholinesterase  and paranltrophenol excretions
              observed in an aircraft loader 
-------
    Figure 3.  Comparisons of cholinesterase and paranltrophenol excretions
              observed in an aircraft loader  (K.D.T.), Belle Glade, Florida 1973
   PPM
A pH/hour

 2.00 r
 1.50
 1.00
 0.75
 0.50
 0.25
                                                     RBC——
         6  28  11  12  13   14  15  16  17  22  23  24  26  28  28  29  30
        =EB.    MARCH                                      APRIL
                                      DATE

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 These figures are presented merely to indicate the interpretative complexities
 that were first encountered when urines were obtained from persons whose pesti-
 cide exposures were multiple and where the data was from Individuals who re-
 ceived pesticide exposure dally.   In addition they illustrated  the need  first
 to obtain baseline information on the magnitude and duration of the urinary
 metabolite excretion following a single exposure to a single pesticide so that
 information could be obtained of the duration of excretion of the metabolite
 and the nature of the diurnal fluctuations.

 In order to better understand some of these,  dally urines were  collected from
 an aircraft loader (Maynard W.).   This man had overt symptoms of anticholines-
 terase intoxication which had developed from occupational exposure to parathion.
 He was hospitalized and  given atropine (this point is shown in  Figure 4).   His
 chollnesterase levels at that point in time  were RBC ChE 0.09 and PI.  ChE was
 0.13ApH/hr;  during the  next month he mixed  and loaded parathion 8E and  ethyl-
 methyl parathion 6-3 mixtures on the following days:

    March 13th,  18th,  19th,  20th,  21st,  23rd,  24th,  27th  and 28th

 He was unable to give us his pesticide exposure on the other days during this
 month.   Figure 4 illustrates that based upon the combined di-ethyl and di-
 methyl alkyl  phosphate levels in  dally grab  samples there was Indeed signi-
 ficant pesticide exposure to ethyl methyl  yet  at the  end of the study period
 a  repeated  cholinesterase showed  that the  red  cell  had risen from 0.09 to  0.35
 pH/hr and  the plasma had risen  from 0.13  to 0.32ApH/hr.

 One observation which may prove  significant  in future studies concern the
 ratio  of DETP to DEP,  as already  mentioned, we have noted that  the loaders
 were more exposed than the  pilots and since  the predominant organophosphate
 exposure was  to ethyl-methyl parathion  these would  be reflected  by the excre-
 tion levels of  the  di-ethyl alkyl phosphate metabolites.   Both  pilots  and
 loaders were  representative and reflective of  persons chronically exposed
 to this pesticide and  we noted that  66% of the  urines from  the  loaders had
 higher  DETP levels  than  DEP  levels whereas the  urines  from  the  pilots
 only 39% had  higher DETP levels than  DEP.  Very useful quantitative Informa-
 tion was also  obtained from the study of the phenolic  derivatives  utilizing
 the  Shaflk et  al. multi-residue procedure.  Paranltrophenol was  identified
 In all  of the urines of  the  pilots and loaders  which means  that however ex-
 amined  these high and  intermediately  exposed group  of people  always had
 qualitative evidence of  parathion exposure.  Trace  amounts  of this parathion
 metabolite were also identified in the mechanic  in  7  out  of 9 specimens.  Penta-
 chlorophenol  (POP)  traces were found  in all 87  urines; the mean concentration
 was  0.01 ppm with a range of 0.004-0.024 ppm.   These  indicate minimal  inciden-
 tal  exposure to this wood preservative and these may be indicative of  general
 population exposure rather than work exposure.

The  identification of 2,4,5-trlchlorophenol was probably reflective of inciden-
 tal  exposure to Ronnel since there was no history of  this insecticide being
used by Allied Helicopters Inc.  Similarly, 2,4-DCP and 3,5,6-TC pryidinol
which are phenolic derivatives of VC-13 and Dursban and must have reflected
 incidental exposure to these pesticides in a domestic setting or even in the
ambient air (as will be seen in another section of this report both insecti-
cides have been regularly identified in the air sampling program).
                                     15

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   Figure 4 - Combined alkyl phosphate excretion in an aircraft loader
             (Maynard W.) during ethyl-methyl parathion exposure.
   PPM
1.0
0.8
0.6
0.4
0.2
                      TOTAL Me PARATHION  EQUIVALENT =  5.3  ppm

                      TOTAL Et PARATHION  EQUIVALENT  = 9.4 ppm
          ATROPINE
        i    i    i    i   i
       6   28  7   11   12  14   16  22  23  24  25  26  27  28
      FEB.     MARCH
                               16

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Thus  the  combination of data from  the urinary alkyl phosphates and  the phenolic
derivatives provide highly  informative qualitative data of the occupational and
incidental exposure of the  agricultural worker, and qualitatively indicate the
potential of the multi-residue surveillance of different occupational groups.
Quantitative interpretation is much more complex as will be described hereinafter.

Discussion - The ultimate long term goal of these studies is to try and find
qualitatively and quantitatively a urine profile of the di-alkyl phosphates
which is  prognostic of excessive exposure, significant cholinesterase inhibition
and incipient cholinesterase inhibition.  If a screening test can be identified
which can be used to monitor the pesticide exposed worker than pesticide handl-
ing can be put on a more rational basis and the worker and Industry will have
the potential for a routine industrial hygiene program.  It is our hope that
the urinary metabolite will become the ultimate occupational surveillance para-
meter.  The studies reported so far have confirmed, a) the potential of the di-
alkyl phosphates and phenolic derivatives as diagnostic indices of organophos-
phate and carbarnate pesticide poisoning.  Collectively the can provide very
specific  Information as the nature of the intoxicant in acute poisoning, b)
they have also been shown to be very useful toxicological monitors during an
acute poisoning. They provide  the essential information as to the degree of
the organophosphate intoxicant on any point in time during the acute phase
in Illness and they provide toxicological data as to when specific antidotes
can be discontinued.  This  potential is ill understood and ill recognized by
physicians having to handle long drawn acute organophosphate intoxications.
c) based  on a small sample  the di-alkyl phosphates appear to have good correla-
tion with cholinesterase Inhibition and these findings confirm Shafik's earlier
observation, however, much more information is needed on the dynamics of these
metabolites under chronic exposure before there is any likelihood of identifying
a quantitative or qualitative profile which is sufficiently indicative of ex-
cessive exposure that can be relied upon as an-occupational surveillance index.
Perhaps,  it will be the DETP-DEP ratio under chronic or occupational exposure
to parathion which might provide the clue.

In order  to explain the next thrust of our studies next year it is perhaps
timely to list some of the unknown variables, these include:

             1.  The duration of metabolite excretion following a single
                 non-toxic exposure to a single pesticide.
             2.  The effects of a single exposure to multiple pesticides.
             3.  The quantitative and qualitative urinary profiles of
                 non-occupational exposure, i.e., how many of these meta-
                 bolites,  if any, can be detected in the general popula-
                 tion—does food, house dust or the use of insecticide
                 strips or  sprays in the home environment result in the
                 pesticide excretion?

It is to these unknown areas that we will turn our study design and the remainder
of this report describe single exposures to single and multiple pesticides.

The first group to be studied was a group of six structural pest control opera-
tors working in the Dade County Housing and Urban Development Program.   Two
of these  (C.L.  and M.H.)  were sprayers, and the four others were maintenance
men and so were peripherally exposed to pesticides.   This group was first stu-
died while working with Baygon (propoxur).   A 1.5% Baygon solution was used
during a period of 8 hours.   Grab samples of urine and blood for cholinesterase
                                      17

-------
 levels were obtained to see if  Iso-propoxyphenol (IFF)  could  be identified  by
 the Shaflk et al method in this type of  exposure.   The  chollnesterase and
 Iso-propoxyphenol are shown in  Table 4,   These preliminary findings confirmed
 that IPP could be readily identified in  this  type  of  work exposure.   The name
 group was similarly tested after they switched from a 1.5% Baygon solution  to
 a 2% Dursban application.   Blood cholinesterases  and eight sequential urines
 were obtained from three spraymen (C.L.,  M.H.,  and M.K.).   Following the switch
 to Dursban,  the average plasma  cholinesterase was  noted to have declined by
 50% though the average red blood cell levels  remained unchanged in the group.
 3,5,6-TC-pyridinol,  the phenolic metabolite of Dursban  was readily identified
 both during and after this occupational  exposure.   Urinary concentrations
 ranged from 0.29-1.03 ppm.   Peak concentrations were  reached  between nine
 and twelve hours after exposure which suggested that  this  might be the optimal
 time for surveillance by this technique  if  grab urine samples were to be used.

 This information prompted  us to try  and  identify structural pest control opera-
 tors with single pesticide exposures and  whose work practices promoted the
 study of pre*exposure baseline  urines.  Ve  have also  sought operators who
 could be studied on the basis of single work  exposures%   Eyeready Pest Control
 Company was  a firm who employed two  operators whose work practices fulfilled
 these experimental design  requirements.   Each operator  worked six to eight
 hour shifts  on Saturdays only;  spending  the rest of the week  attending college,
 Both agreed  to participate in the study and to provide  sequential urines,

 The first study was  a Baygon^  study.  Pre-exposure  urines and bloods were
 collected.   Max P.  used a  hand  spray and  Edward S.  a  power spray.   Ed wore
 rubber gloves and respirator; both mixed  their  Individual  spray solutions
 reducing a 14% Baygon concentrated solution to  a 0.5% solution for applica-
 tion.   They  sprayed  from 8:00 a.m. to 4:00  p.m.  The  sequential IPP  excretion
 over this period is  shown  in Table 5 and  Figures 5  and  6.

 Results - Firstly the men  had negative urines at the  commencement of  the study
 thus confirming their non-exposure history  of the previous day.   Secondly,
 Max  sustained more exposure  than had Edward,  a  fact which  may be attributed
 to  the  different amount  of exposure  from  the  hand spray  versus the power spray
 and  also the  use of  protective clothing.  Thirdly,  in both the peak  exposure
 time occurred five hours after completion of  work.  Fourthly,  in one  spray
 operator (Max P.), his excretion of  the metabolite was completed within  16  hours
 after  his last  exposure; in  the  other sprayman  (Edward S.)  36 hours was  in-
 sufficient time for  the  total excretion of  the metabolite,  trace amounts
 still being detected  at  the  end  of this time.  The  total amount  of IPP excreted
 in  the  30 hours  for  Max was  1,550  ug which  is equivalent to 2,139  ug  of  Baygon.
 In  the  other,  Edward,  S.,  712 ug  of  IPP was excreted  in  29  hours  which  is the
 equivalent of  982 ug  of Baygon.    It  is possible  to relate  this metabolite's
 excretions to  the original exposure  of the  intact pesticide.   In order  to
 obtain  a  rough  estimate of the percentage of  the Intact  pesticide  that is
 applied  the micrograms of IFF are multiplied by a factor of 1.38  to convert
 them to  the intact pesticide of Baygon based on  their molecular weight.

 A second  thirty  six  hour study was conducted  in  these spraymen  following
 their single  4*j  hour  exposure to 2^% Dursban and 0,75% of DDVF«  Urine levels
 of DMP would be  indicative of DDVP exposure and Dursban would be  identified
 by DEP and DETP  alkyl  phosphate excretion and also by 3,5,6-TC prydinol
 excretion.  The  excretion pattern of these metabolites in  the  two  spraymen
 are shown in Table 6  and Figures 7 and 8.  Here again several  interesting
 findings were noted.   Firstly,  excretion of  the metabolite from Dursban and
DDVP were still  occurring 36 hours after this single 4% hour exposure. Secondly,
 the maximum excretion of DMP was noted 6 to 9 hours after exposure.  Thirdly,
Max never excreted DEP in contrast to Edward.   This was also observed in a  sub-

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Table 4.  CttOLINESTERASE AND URINARY ISOPROPOXYPHENOL LEVELS IK BAYGON
          (0.5%) IN STRUCTURAL PEST CONTROL OPERATORS AND ASSISTANTS
Identification
C.L.
M.H.
H.L.
C.J.
R.D.
Work
classification
Spray operator
ti ii
Maintenance
ii
ii
Cholinesterase
pH/hr
RBC Plasma
0.96
0.99
0.90
0.89
0.72
1.04
1.33
0.91
0.90
0.98
Iso-propoxyphenol (ppra)
7.99
0.98
0.32
<0.10
0.11

-------
fO
o
Table 5. TWENTY FOUR HOUR EXCRETION OF ISOPROPOXYPHENOL IN TWO SPRAYMKN
OCCUPATIONALLT EXPOSED FOR SIX HOURS
Time of
Operator Voiding
Max P. 8:15 a.m.
12:30 p.m.**
4:10 p. .
6:05 p. .
9:30 p. .
11:30 p. .
8:00 a. .
2:30 p. .
Edward S. 8:20 a.m.
3:00 p.m.
9:45 p.m.
3:28 a.m.
1:00 p.m.
IPP (ppm)
UK/ml*
N.D.
0.07
0.26
0.63
2.3
2.5
0.56
0.12
N.D.
0.10
2.4
0.54
0.56
TO 0.5Z BATGON SPRAT SOLUTION
IPP
ue/hour
N.D.
1.69
18.38
56.6
249.1
149.4
22.8
3.5
N.D.
4.8
70.8
5.9
16.6
Baygon (ug/hour)
equivalent
N.D.
2.33
25.4
78.1
343.8
206.2
31.5
4.8
N.D.
6.6
97.7
8.1
23.0
         Corrected for 800 mOsm/1
         **exposure period

-------
Figure 5 - OriBaryKBXctetion of Xso-propoxyphenol (Baygon 0.5%)  in a structural
          pest control operator (Max P.) following six hours of exposure.
   350
   300
   250
   200
   150
   100
    50  •
     Jjg  IPP/HOUR
;2>- jug  BAYGON
     EQUIVALENT/HOUR
                                                                 j
      8:30   12:30  4:10  6:05 9:30 11:30
       AM     PM
                                          HOUR
        8:00
        AM
2:30
 PM
                                21

-------
Figure 6 - Urinary excretion of Iso-propoxyphenol (Baygon 0.5%) in a
           structural pest control operator (Edward S.) following six
           hours of exposure.
100
80
60
40
20
0
1
-



-©- jug IPP/HOUR
-©- jug BAYGON
EQUIVALENT/HOUR
^ /•>

i i i
8:20 3:00 9:54 3:28 1:00
AM PM AM HOUR PM
1-3-73 11-4-73
                                   .-2

-------
                        Table 6.   THIRTY-TWO HOUR URINARY EXCRETIONS OF 3.5,6-TC PYRIOINOL AND
                                  BMP,  DBP.  DETP  IN TWO STRUCTURAL PEST CONTROL OPERATORS
to
Operator
Max P.









Sdward S.








Time of
Voiding
8:15 a.m.
2:45 p.m.
6:20 p.m.
8:00 p.m.
9:45 p.m.
11:45 p.m.
12:30 a.m.
8:42 a.m.
10:50 a.m.
1:30 p.m.
8:27 a.m.
2:49 p.m.
6:35 p.m.
11:00 p.m.
5:00 a.m.
8:20 a.m.
3:45 p.m.
6:25 p.m.
8:00 p.m.
3,5,6-TC
Pyridinol
ue/hour
N.D.
1.1
2.1
3.4
2.4
3.0
3.3
2.7
3.2
2.2
N.D.
2.4
3.5
4.1
3.7
4.3
4.1
7.1
7.5
Dursban
equivalent
N.D.
1.95
3.72
6.02
4.25
.5.31
5.84
4.78
5.66
3.89
N.D.
4.25
6.20
7.26
6.55
7.61
7.26
12.6
13.3
DMP
ue/hour
N.D.
5.6
19.9
20.7
20.1
16.1
11.1
5.1
Tr.
N.D.
N.D.
9.5
33.9
29.0
14.7
6.2
5.8
6.8
7.1
DEP
ue/hour










N.D.
N.D.
N.D.
2.1
1.2
1.7
2.0
3.3
2.8
DETP
ue/hour
N.D.
8.5
9.6
16.9
16
11.0
13.8
9.6
12.4
3.8
N.D.
1.4
9.1
11.0
8.6
4.8
1.3
9.3
11.2

-------
 Figure 7.  Comparisons of urinary  3,5,6-TC pyridinol metabolite and alkyl-
            phosphate metabolites observed in a structural pest control
            operator (M.P.),  Miami, Florida 1973
20.0 r
15.0  -
10.0  -
 5.0  -
   0L
-
-



- .3— J""l_
r2
F— 1— J •— '
-Q> 3,5,6-TC
PYRIDINOL
-<7>- DETP
.3. -<3> DMP
-fl n



1 1 1 1 1 1 1 II
-S
1
8:15 2:45 6:20 1 9:45 12:30 AM 8:42 10:50 1:30
AM PM 8:00 n:45 HOUR PM
      11-17-73
11-18-73
                                    2A

-------
Figure 8.   Comparisons of urinary  3,5,6-TC pyridinol metabolite and alky I
           phosphate metabolites observed in a structural pest control
           operator (E.S.),  Miami, Florida 1973
                                                       3,5,6-TC
                                                        PYRIDINOL

                                                       DETP
                                                       DMP
                                                       DEP
   8:27
   AM

11 -17-73
2:49
 PM
                     6:35    11:00
 5:00
  AM

11-18-73
8:20
3:45 6:25 8:00
 PM
       HOUR
                                 25

-------
sequent experiment not herein reported; it was found that once again the same
sprayman (Max) had DETP but no DEP.  Whether this reflects an inherent enzyme
defect is conjectural and is under further study.  Fourthly, maximum excretion
of DETP was 6 to 9 hours after exposure but 3,5,6-TC pyridinol reaches a peak
much later.  Fifthly, both men had detectable amounts of these urinary meta-
bolites even after 36 hours of study.

The Dursban and DDVP equivalent of the total exposure of the total exposure of
each sprayman was calculated on the basis of the di-alkyl phosphate metabolites.
In Edward S., 211 ug of DETP was excreted in 36 hours which is equivalent to
357 ug of Dursban.  In addition, 50 ug of DEP was excreted in 36 hours which
is equivalent to 91 ug of Dursban.  Together, therefore, this alkyl phosphate
metabolites reflect a total exposure of 448 ug of Dursban.  In Max P., 293 ug
of DETP only were excreted which is equivalent to 496 ug of Dursban.  Thus,
on the basis of the di alkyl phosphate the mono alkyl phosphate cannot be
measured as such, therefore the calculated intact pesticide is perforce of an
accured expression of the total exposure of these workers to the pertinent
pesticide.
                                 26

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                                 SECTION V

                           AIR SAMPLING STUDIES


 B.  Air  Sampling - Background Information

 While  the routine monitoring of  air for pesticides and other environmental
 pollutants has assumed increased importance, available information on the
 qualitative and quantitative profiel of air pollution has suffered from in-
 adequacies in the air samplers being used, earlier analytical procedures and
 the number and location of existing sample sites.  In-house research in the
 Environmental Protection Agency  had addressed themselves to these problems on
 chemical analyses and improvement in air sampling techniques and have made
 significant progress in both areas.  A new analytical method needed to be
 evaluated and the introduction of a double impinger system into the air
 sampler needed to be tested.  The objectives of this project sought to evalu-
 ate these new approaches.  First study were conducted in the laboratory to
 check  the recovery of 25 pesticides, and secondly in the field-to evaluate
 and test the double impinger sampler system in different locations in South
 Florida.

 Description of the various phases -

    Pursuant to the objectives,  the following phases were planned:

    Phase I - In-house training  of the chemists in new methodologies.

    Phase II - To measure the percent recovery of 25 pesticides in a
               double impinger system.

    Phase III - To measure trapping efficiency and percent recovery of
                25 pesticides in ethylene glycol.

    Phase IV - To field test the equipment in three different locations
               of South Florida.

Materials and Methods

 In order to evaluate a new analytical method an investigation of several sub-
strates (cottonseed oil,  etc) were first analyzed.  Cottonseed oil, silica
gel and ethylene glycol were the materials selected to assess the trapping
potential of each.   Duplicate experiments were also conducted with ethylene
glycol.  50 ml.  of ethylene glycol was placed in each impinger, the first im-
pinger was spiked with 25 pesticides and the percentage recovery of each
pesticide in each Impinger measured.  The second experiment tested the trapping
potential of ethylene glycol by drawing ambient air through a U shaped tube
in an oil bath spiked with the same 25 pesticides.  Air was drawn in at 0.82
cubic feet per minute for a 12 hour period giving a total volume of 16.4 m  of
air.   After 12 hours the pesticide concentrations in each impinger were ana-
lyzed.   Midwest Research Institute air samplers were placed in a Miami sub-
urb,  at the Miami International Airport and at a remote site in the Everglades,
40 miles west of Miami.   A double Impinger system was used in each sampler and
100 ml. of ethylene glycol placed in each impinger.   During the first 12 hour
period, air was drawn through the first double Impinger system and at the com-
pletion of this 12  hour period,  a second sample of air was drawn through the
second double impinger system.   The vacuum pump was run at full capacity during
the 24 hour period.   The 24 hour  volume of air was recorded as was the wind
direction and wind velocity.

                                    27

-------
 This  24  hour  sample was  transferred  to  the University of Miami  laboratory
 and analyzed  without  delay.   The  analytical method used to measure  these
 air samples was  by the Sherma and Shafik method.

 Results

 Ethylene glycol  was found  to  be the  most suitable trapping material.  The
 results  of retention  of  25 pesticides using ethylene glycol  following a 12
 hour  run are  shown in Table 7.  Table 8 describes the trapping  efficiency
 of ethylene glycol for the 25 pesticides.  Table 9 presents  the air  concen-
 trations of pesticides identified in the three  sampling sites.  Simultaneous
 samplings were done at the Everglades and suburban sites.

 Discussion

 The reproducibility of pesticide  recovery in both spiked samples  is  testimony
 of the reproducibility of  the analytical method.  Similarly, maximum entrap-
 ment  occurred in the  first impinger  and in the  second impinger  between 12 -
 22% of the various pesticides trapped.  The types of the several  pesticides
 identified in the various  ampling sites emphasized the changing profile
 of ambient pesticides in these areas.   It will  be observed that VC-13 was
 identified on four occasions  in the  Miami suburbs, as also was  Dursban.
 These pesticides are  currently being extensively used in homes  and gardens
 and their presence is illustrative of predominant organophosphate profile
 of the Miami  area.

 The qualitative  profile  of the pesticides identified is strikingly different
 to air sampling  studies  conducted in previous years by virtue of  the pre-
 dominance of  minimal  traces of these new type of organophosphates in con-
 trast to the  usual presence of the organochlorine pesticides. The following
 pesticides were  not detected  in the  s,even samples sites:  p,p'-DDT,  o,p'-DDE,
 B-BHC, HCB, Endrin, Trithlon,  Ethian^   Rpnnelf Pa.rathion,  a.nd methyl parar
 thion.   DDT was  not found  in  any  sample though p,p'-DDE was still present in
 trace amounts.   The traces of malathion are almost certainly reflective of
mosquito  control practices.   The  Everglades sampling site which is 40 miles
away  from the two other  sites  in  the downtown Miami area and the  nearest
agricultural area is more  than 20 miles away, the preponderance ofoCBHC
related  to 1 BHC in this site  is  difficult to explain.   The identification
of Dursban in the Everglades  is most surely reflective of aerial  transport
from  the urban coastline areas since Dursban is not agriculturally applied.

In addition to the 20 pesticides  listed in Table 9,  VC-13 and Dursban were
Identified in the second fraction; the percentage recovery through silica
gel of these two pesticides were 99 and 100% respectively.
                                  28

-------
Table 7.  RETENTION OF PESTICIDES IN ETBTLEHB GLTCOL
             (12 hr. run-Flow .82 CFM (16.4 n3)
Concentrations In
nanoftrans
840
1620
2070
3765
2400
545
2300
80
100
200
600
800
800
80
801
180
400
800
200
200
400
800
800
400
400
Pesticide
Sonnel
Methyl Parathlon
Ethyl Parathlon
Trithlon
Ettiion
Dlazlnon
Malathion
BHC
Aldrln
pp« DDE
op DDT
pp1 DDD
pp« DDT
Llndane
B BHC
Heptachlor Epoxide
Dieldrln
End r in
Baygon
2,3,5-Landrin
Carbofuran
Matacil
Zectran
Carbaryl
Meaurol
Percent recovery
S
Impinger 1
73
85
91
93
96
88
97
62
23
94
84
80
90
91
79
81
96
93
70
74
73
69
65
99
85
S
ImplnRer 2
11
Tr
Tr
Tr
Tr
7
Tr
21
39
10
Tr
__
..
11
__
10
Tr.
Tr.
Tr.
16
__
10
—
~

s
Total
84
85+
91+
93+
96+
95+
97+
83
62
104
84+
80
90
102
79
91
96+
93+
70+
90
73
79
65
99
85
Percent recovery
S2
Impinger 1
76
93
96
97
102
87
94
70
17
81
81
82
89
97
81
82
98
93
88
83
83
77
70
98
98
S2
Impineer 2
10
Tr
Tr
Tr
Tr
8
Tr
22
38
11
Tr
__
__
7
__
7
Tr
Tr
Tr
10
.._
10
__
__

S2
Total
86
93+
96+
97+
102+
95
94+
92
55
92
81+
82
89
104
81
89
98+
93+
88+
93
83
87
70
98
98

-------
                               Table 8.  ETHYLENE GLTCOL TRAPPING EFFICIENCY
                                        (12 hr. run=Flow .82 CFM (16.4 m3 of air)
Concentrations in
U-tube (ng)
840
1620
2070
3765
2400
545
2300
80
100
200
600
800
800
80
801
180
400
800
200
200
400
800
800
400
400
Pesticide
Ronnel
Methyl Parathion
Ethyl Parathion
Trithion
Ethion
Diazinon
Malathion
(XBHC
Aldrin
p,p'-DDE
o,p'-DDT
p,p'-DDD
p.p'-DDT
Lindane
B-BHC
Heptachlor Epox.
Dieldrin
Endrin
Baygon
2,3,5-Landrin
Carbofuran
Metacil
Zee t ran
Carbaryl
Mesurol
Percent Recovery
AR
Impineer 1*
52
53
63
53
58
46
56
43
23
74
71
73
73
59
53
64
73
78
38
40
40
26
18
60
49
AR
Impinger 2*
19
15
17
13
14
16
16
22
22
20
17
16
16
20
15
21
17
18
18
21
16
12
Tr
13
12
Total*
71
68
80
68
72
62
72
65
45
94
88
89
89
79
68
85
90
96
56
61
56
38
18+
73
61
U-
Tube



7%
12%




















Percent Recovery
AR2
Impineer 1
62
76
78
74
77
68
73
64
17
78
75
78
81
96
64
67
77
78
65
65
65
44
28
91
73
AR2
Impineer 2
23
16
18
16
17
18
14
28
30
25
19
19
19
25
11
20
16
18
20
24
16
• 17
TR
19
15
AR2
Total
85
92
96
90
94
86
87
92
47
103
94
97
100
111
75
87
93
96
85
89
81
61
28+
110
88
*(4 hour power failure)

-------
           Table 9.   AIR CONCENTRATIONS OF PESTICIDES (ng/m3) FROM DIFFERENT SAMPLING SITES IN SOUTH FLORIDA 1973
Pesticide
VC-13
Dursban
Dlazlnon
Malathlon
oCBUC
Heptachlor
Aldrin
Lindane
Dleldrln
p,p'-DDE
M3 Collected
Wind Direction:
Start
Finish
Suburban
Sample f 1
ng/m3
1.09
1.68
1.02
____
0.51
1.00
0.25
0.68
0.40
0.18
43.15

N-NE
S-SE
Sample $2
ng/m3
1.21
1.82
0.78
_____
0.65
0.52
0.13
0.41
0.35
____
48.92

SW
NW
Sample f 3*
ng/m3
0.76
2.04
2.27
7.24
0.95
0.51
0.18
0.93
0.42
— —
45.87

NE
N-NE
Sample 14*
ng/m3
0.63
1.47
0.30
____
1.37
0.58
0.10
0.62
0.46
0.19
53.71

ESE
SE
Everglades
Sample $5*
ng/m3
	
0.87
	
____
0.85

	 	
0.18
0.18

45.81

NE
NW
Sample 16*
ng/m3
	
0.77
0.30
— —
1.32

—.-_
0,19
0.20
— —
47.74

E-NE
E-SE
Miami Airport
Sample 87
ng/m3
2.95
6.10
1.00
1.68
0.97
2.65
0,89
0.42
0.67
0.62
49.94

SE
NW
*slmultaneous samples ('3 and 5, #4 and 6)

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                                SECTION VI

                               EEC STUDIES


C. EEC Studies - Scope, purpose and background information

Changes in the electroencephalograph following human pesticide exposure are
dependent upon the type of pesticides Involved and the nature of the exposure.
In acute poisoning, the duration of cerebral anoxia has been shown to produce
EEC changes.  Hypothalamic spikes were noted by Holmes which persisted three
years after a parathion exposure.   Brown reported persistent EEC changes simi-
lar to those seen in temporal lobe epilepsy following mild acute organophosphate
exposure.   Metcalf and Holmes reported unusual EEC changes in organophosphate
exposures.    Hunter and Robinson conducted EEC studies in three groups of
volunteers fed 0.01, 0.05 and 0.211 mg of dieldrin per man per day, no abnor-
mal EEC changes were noted with this amount of dieldrin intake.    EEC changes
were prognostic of incipient convulsions due to over exposure to the diene
group of insecticides.  The occurrence of this degree of incipient toxicity
is nowadays avoided by relying entirely on threshold blood levels of dieldrin.

Animal studies recently conducted in the U.S. Environmental Protection Agency
Perrine Primate laboratory by Dr. John Santolucito and others in the Pharmaco-
logical Branch have compared chronic and low level exposure effects of Carbaryl
on the EEC of monkeys.  Similar abnormalities have been noted following para-
thion, dieldrin and DDT exposure.  Using a portable EEC machine suitable for
battery operation in the field it was planned to conduct EEC studies in human
volunteers who were occupationally exposed to pesticides.

Description of the various phases

The first phase was the completion and in-house evaluation of the portable EEC
apparatus.  The second phase was the training of project personnel in EEC
techniques and a medical review of the equipment and its application by the
Division of Electroencephalography, University of Miami School of Medicine.
The third phase was the implementation of any necessary modifications suggested
by this Division and pre-testing on non-exposed volunteers, and the fourth and
final phase, field application of the machine.

Materials and methods

The EEC machine to be used is a portable battery operated apparatus with facil-
ities for recording tracings on a tape unit.  There will be digital conver-
sion of electronic recordings which are transcribed from electro-magnetic
tapes.  These will be submitted to Dr.  Santolucito in Research Triangle Park
for interpretation.   It'was planned to use intracuticular scalp recordings
from needle electrodes rather than surface electrodes.  These should dimish
the background interference.   After obtaining signed consent forms, the tech-
nique to be used in testing was as follows:   Three sterile disposable butterfly
infusion needles would be inserted at three sites into the following areas of
the scalp after cleaning this area with acetone.   One anterior electrode placed
1*5" posterior to the nasion,  a second left lateral electrode 14" to the left
of the occipital, and third,  a right lateral electrode place IV to the right
of the occipital point.  The subject would be examined in a recumbent position
with the eyes closed and tracing would be run for a fifteen minute period.
                                    32

-------
Interim Report

Prior to the receipt of the portable unit in the second half of the project
year, the field investigator received instructions on electroencephalographic
techniques from the Division of Electroencephalography in the Department of
Neurology of the University of Miami School of Medicine.  The director of
this Unit, Dr. E.T. Richey and his Blomedlcal Engineering representative re-
quested a review of the specifications of the apparatus and agreed to review
the procedure.  Following a preliminary run on a non-exposed human volunteer,
Dr. Richey expressed concern with regard to low input impedence and questioned
whether the electrical tracing from the three scalp sites could be interpreted
with ease.  It was not understood at this time that the frequency rather than
the actual wave patterns were being measured.  Questions were also raised as
to whether there was any hope of altering the technique to obviate the necessity
of intracuticular electrodes in favor of using the conventional and acceptable
surface electrodes.  It was agreed to provide the EEC Division with the speci-
fications of the apparatus from Biomedlcal Engineering, and to arrange a con-
ference with Dr. Santolucito prior to moving to phase 4 - the field applica-
tion.  Additional safety modifications suggested by the Director of the Division
of EEC have been Implemented and'intracuticular electrode will still be used.
The machine is now ready for use in the field.
                                 33

-------
                                SECTION VII

                 ACUTE PESTICIDE POISONING INVESTIGATIONS
D.  Acute Pesticide Poisoning Investigations - Scope, purpose and background
    Information - The two objectives of this work unit were:

    1.  To Investigate and confirm suspect pesticide poisoning cases
        occurring in the Dade County area, and to document the clinical
        and therapeutic responses observed at the bedside.  These find-
        ings, with medical confirmation of suspect cases, would be made
        available to the Dade County Community Studies Pesticide Project.

    2.  To study pesticide metabolite dynamics in blood, urine and fat
        in acute poisoning cases hospitalized in the Dade County area
        and to determine the informative potential of these in the clinical
        management of the case.

Although pesticide poisoning has not been a reportable disease in Florida,
the resources and experience of the Miami Pesticides Laboratory have become
known to most hospital Emergency Rooms and pathologlsts in the South Florida
area and so has been widely used.  In the past, epdiemiologic and laboratory
confirmation has been offered through the Community Studies Program and it
was planned in this project to continue this function in the University and
to facilitate acquisition of pesticide poisoning incidence data in the area.

Results

Table 10 lists the confirmed fatal pesticide poisoning cases investigated by
this project and Table 11 lists the number of confirmed non-fatal cases.
Table 12 presents the clinical and analytical features of an additional 26
investigated cases of suspect pesticide illnesses which the project investi-
gated during this year.

In February, 1973, no further support was received at the Florida Community
Pesticide Studies terminated, therefore, the data has been made available to
the Pesticides Operations Office in Washington, D.C. and in the Southwest
Region IV (Drs. Evrard and Kessler) so that the material could be utilized
by the appropriate Federal authorities.  Administratively, therefore, reports
were transferred from Jacksonville, Florida to Atlanta, Georgia.

Discussion

1 - Confirmed fatal cases - All three fatalities were due to organophosphate
    exposure; these pesticides were identified in the various specimens and
    are shown in the table and all were confirmed by gas liquid chromatography
    in the flame photometric and electron capture detectors.  One of the deaths
    resulted from the ingestion of VC-13.   This pesticide is becoming an in-
    creasingly serious chemical problem in the area and the material is readily
    available in most garden stores.   Illness following ingestion of this
    material is slow to evolve and the clinical picture is protracted in many
    instances.   In the past,  the patient has Been hospitalized initially more
    of a precautionary measure and then what usually happens is that quite
    suddenly a severe and life threatening cholinergic crisis will develop
    calling for heroic antidotal and  resuscltative management,  The danger of
    this type of intoxication lies in the  deceptively mild clinical picture
    at onset,  a feature  which might tempt  the inexperience to discharge the


                                   34

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                      Table 10.  CONFIRMED FATAL INTOXICATIONS IN SOUTH FLORIDA, DECODER 1972-DECEMBER 1973
01
Name Race/Sex Age
1. Juan M.

2. Henry K.






3. Walter M











W/M

W/M






W/M











20 y/o

76 y/o






51 y/o











Date of Type of
incident Exposure Material Confirmed clinical and laboratory tests
12/31/72

5/9/73






9/7/73











Ingest ion-
Suicide
Ingest ion-
Suicide





Ingest ion-
Suicide










Parathion

VC-13






Diazinon 51Z











RBC ChE 0.15 PI. ChE 0.15
Parathion in blood 18.3 ppb.
Gall Bladder - VC-13 - 37.5 ppm. by F.P.D.
Fat from kidney - VC-13 - 174 ppm. by F.P.D.
Urine = 2,4-DCP (chloroacetate derivative) 12
Liver - VC-13 - 3.6 ppm. by F.P.D.
Gastric content (294 mg. bloody) - 0.34 ppm
by F.P.D.
Whole blood - VC-13 - 1.0 ppm by F.P.D.
9/10/73 - RBC ChE 0.20 PI. ChE 0.07
Pre PAM- Diazinon in blood 34 ppb. by EC & I
9/10/73 - RBC ChE 0.25 PI. ChE 0.06
Post PAM - Diazinon in blood 17 ppb. by "
9/12/73 - RBC ChE 0.29 PI. ChE 0.09
Diazinon in blood 9 ppb. by EC &
9/13/73 - RBC ChE 0.30 PI. ChE 0.09 by " &
Diazinon in blood 10 ppb. E.G.
9/18/73 - RBC ChE 0.32 PI. ChE 0.09
Diazinon not found (FPD & EC)
p,p'-DDT <2, p.p'-DDE 7.1 and Dieldrin <1 pp
respectively .

-------
 Che patient home If the probabiluty of a delayed onset of illness is not
 appreciated.  The halogenic portion of this organophosphate pesticide
 favors deposition in fat, and this is the same reason for the slow evolu-
 tion of symptoms.  The fat solubility potential also calls for the daily
 monitoring of blood for VC-13 and urine for the phenolic metabolite 2,4-
 DCP.  Atropine therapy must be continued as long as the parent compound
 or its urinary metabolite can still be identified, and atropinization
 may have to be continued for several days.  Experience gained in this
 special type of organophosphate exposure (halogenated organophosphate)
 emphasizes the need for further study of acute and chronic exposure to
 organophosphate pesticide that have fat soluble characteristics.

 2 - Confirmed cases - Incidence Data ~ Eighteen non^fatal pesticide poison-
 ings were confirmed in 1973.  The types of poisonings, materials involved
 and results of the appropriate analytical diagnostic tests are shown in
 Table 11.  Eighteen cases were due to organophosphate intoxications and
 ten of these were from occupational exposure.  In contrast to residue
 problems in California, once again in South Florida, our investigations
 have failed to confirm an episode of "picker poisoning", a topic of
 current discussion of political importance and exhibiting succinct
 geographical distribution characteristics.  Arsenic and pentachloro-
 phenol were the other agents Involved besides the organophosphate insecti-
 cides.

 The case of arsenic poisoning occurred in a 6 year old white male child
 who was poisoned by some toys that he picked up in a garbage dump.  Weed
 killer had been discarded into the dump and has spilled over the toys.

 The pentachlorophenol intoxication was observed In a carpenter and the
 diagnosis was made on the basis of exposure, clinical picture and urine
 results which were obtained eight weeks after his last exposure.  The
 patient was a 60 year old white male carpenter who had had extensive
 exposure to pentachlorophenol which he used for termite control.  Besides
 complaining of weakness  anorexia and loss of weight, the patient had
 severe dermatitis with fissures of the skin.  He also developed an exten-
 sive generalized and painless lymphadenopathy which resembled Hodgkins
 Disease.  Lymph node biopsy, however, did not confirm this diagnosis,
 the section showing only generalized chronic inflamatory changes.

 Discussion 3 - Unconfirmed Cases - Twenty six cases of pesticide associated
 illnesses were investigated, and on the basis of clinical findings, ex-
 posure histories, pathological findings and appropriate laboratory ana-
 lyses were considered not to be due to pesticide exposure.  Five of the
 cases were fatal, and at the time of death, health or agricultural offi-
 cials suggested suspect pesticide exposure.  The definitive diagnosis
 was not made available to us in all cases but alcoholic seizures, acute
 asthma with chronic obstructive lung disease, methaqualone overdose and
meningitis were the ultimate causes of death in four of the five uncon-
 firmed cases.   Of significant medico-legal importance was the occurrence
 of an acute asthmatic attack in two agricultural inspectors, both had
 been employed by the Florida State Department of Agriculture for many
 years and both had a history of chronic bronchitis, emphysema and periodic
 attacks of asthma.   One of the Inspectors died after inspecting a celery
 field; it was first thought that the cause of death was the result of
 parathion residues on celery or an exposure from the drift from aerial
 applications in the adjoining fields.  Leaf residues were insignificant
 and no paranitrophenol or paraoxon were found in the liver or kidney by
 gas chromatography using the electron capture and flame photometric

                               36

-------
                            Table  11.   CONFIRMED NON-FATAL INTOXICATIONS IN SOUTH FLORIDA, 1973
Race,
Name Sex
1. Abraham W. B/M
2. Frank R. B/M
3. Sammy G. B/M
4. April P. W/F
5. Maynard W. B/M
6. Felix S. Ind./H
7. James C. P.W/H
8. Jeral F.* W/M
9. Fernando B.W/M
0. Charles P. B/M
Age
(yrs)
61 y/o
57 y/o
21 y/o
19 mos.
27 y/o
21 y/o
25 y/o
18 y/o
5 y/o
25 y/o
Date of
incident
1/4/73
2/6/73
2/6/73
3/7/73
3/8/73
3/9/73

3/23/73
3/5/73
3/11/73
Type of
Exposure
Dermal-
Occupational
Occupational
11
Ingest ion-
Accidental
Occupational-
Dermal
Dermal-
Occupational
Ingestion-
Suicide
Oc cupat ional-
Accidental
Ingestion-
Accidental
Occupational-
Dermal
Material \
Phosdrin
Organo-
phosphate.
11
Dursban and
Diazinon
Organo-
phosphate
11
VC-13
Organo-
phosphate
Dematon
Parathion
Confirmed clinical and diagnostic tests
1/4/73 - Pre-PAM - Serum ChE 0.14
Post PAH - RBC Che 0.26 PI. ChE O.E
1/5/73 - RBC ChE 0.16 PI. ChE 0.12
Unable to detect Phosdrin by E.C.
RBC ChE 0.21 PI. ChE 0.49
RBC ChE 0.11 PI. ChE 0.15
Asymptomatic, taken off work
3/8/73 - RBC ChE 0.14 PI. ChE 0.10
3/7/73 - Gastric contents pH 1.72
Diazinon 45 ppm. and Dursban 213 ppm by F.PB
3/8/73 - Serum - Diazinon 0.6 ppm. and
Dursban 0.4 ppm. by F.P.D. and E.C.
3/7/73 - Post PAM RBC Che 0.09 PI. 0.13
3/8/73 - RBC ChE 0.18 PI. 0.16
Hospitalized
90% Inhibition colormetric method
Case confirmed by clinical picture
80% Inhibition Fleisher Pope method
Case confirmed by clinical picture.
RBC ChE 0.48 PI. ChE 1.26 (5/16/73)
Case confirmed by hospital notes and
clinical picture.
us
                               *A second incident occurred on Hay 12th when patient was hospitalized for
                                spilling Phosdrin on akin and inhalation.  Case confirmed by clinical
                                picture and atropine response.

-------
Tohi* 11 (Vr^f-i™,^ CONFIRMED NON-FATAL INTOXICATIONS IN SOUTH FLORIDA, 1973
Name
11. Wesley S.
12. Johnny L.
13. Stephen B.
14. Francisco C
15. William K.
16. Walter B.
17. Donald T.
Race,
a ex
B/M
B/M
W/H
. W/H
W/H
B/M
W/H
A^e
30
46
18
14
60
2
6
Date of
incident
3/3/73
1/8/73
5/9/73
6/20/73
7/2/73
9/10/73
10/17/73
Type of
exposure
Occupational
Dermal
Occupational-
Inhalation
Occupational-
Dermal
Ingest ion-
Accidental
Occupational-
Carpenter
Dermal-
Accidental
Ingest ion-
Accidental
Haterial
Organo-
phosphate
Parathion
Organo-
phosphate
Spectricide
(Diazinon 25%)
Pentachloro-
phenol
Parathion
Arsenic

Case confirmed by clinical picture and atropii
response .
Cholinesterase 13 ChE-tel units (Normal 45-90)
Serum ChE 20 (N 45-90) 1/9/73
RBC ChE 0.35 PI. ChE 0.32
5/10/73 RBC ChE 0.29 PI. ChE 0.34
Pre- treatment (Hospital Notes) ChE .45 (Norma
1.90-3.0) Post-treatment PI ChE 22 units (45-
units Normal range)
6/22/73 - RBC ChE 0.70 PI. ChE 0.08 (Michel
Method)
Serum PCP .513 ppm, Urine 0.3 ppm. (Analyzed
9/21/73 p.p'-DDE 39 ppb. p,p'-DDT 8 ppb.
dieldrin 1.5 ppb.
7/2/73 - RBC ChE 0.65 PI. ChE 0.68
Case confirmed by clinical picture and atropi
response.
9/15/73 - RBC ChE 0.50 PI. 0,72 (Analyzed 9/1
collected at 4:17 p.m.
9/15/73 - RBC ChE 0.62 PI. 0.87 (Analyzed 9/1
collected at 10:16 p.m.
p.p'-DDE 2 ppb., no p.p'-DDT, dieldrin Ippb.
No organophosphate found by F.P.D. and EC
Case confirmed by M.E.'s office
Urine 16 mg/ml. Gut Belt method
Field investigation made.

-------
detectorn.  Other mechanisms involved in unconfirmed pesticide poison-
ings related to 1) the occurrence of dermatitides and 2) to the chance
finding of unusually low red blood cell or plasma cholinesterase.  An
example of the former was the occurrence of erythema multifonne in a
52 year old white male whose home had recently been sprayed with Baygon.
Iso-propoxyphenol was not identified in the urine of the other members
of the family.

With regard to the latter, low cholinesterase values were identified in
a three week old black male infant with pneumonia and possible pesticide
exposure.  In order to evaluate normal physiological occurrence of low
red blood cell and plasma cholinesterase in the newborn, five blood
samples were collected and analyzed from healthy premature babies of
the same birth weight as the case in suspect.  The average red blood cell
of these infants were 0.444pH/hr and for plasma cholinesterase the
average level was 0.47 ApH/hr respectively.

At the request of Dr. Suggs and with Dr. Durham's permission, pseiido-
cholinesterase studies were conducted in individuals with low plasma
cholinesterase as well as their families.  These studies were recon-
firmed by Dibucaine studies by Dr. Suggs in the Center for Disease
Control in Atlanta, Georgia.   A ciguatera poisoning was also confirmed.
                             39

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                                 SECTION vin

                                 REFERENCES
  1.   Shafik.,  T.M., H.C.  Sullivan  and H. F. Enos.  A Multiresidue Procedure
      for Halo and Nitrophenols-Measurement of Exposure to Biodegradable
      Pesticides  Yielding These  Compounds as Metabolites.  J. Agr. Fd. Chen.
      21:295-298, 1973.

  2.   Shafik,  T.M., D. E.  Bradway, H. F. Enos, and A. R. Yobs.  Human Exposure
      to Organophosphorus Pesticides.  A Modified Procedure for the Gas Liquid
      Chronatographic Analysis of Alkyl Phosphate Metabolites in Urine, j. Agr.
      Fd.  Chen.   In Press

  3.   Michel,  H.O.  An Electrometric Method for the Determination of Red Blood
      Cell and Plasma Cholinesterase Activity.  J. Lab. Clin. Med. 34:1564-
      1568,  1949.

  4.   Method for  the Determination of Metabolite or Hydrolysis Products of
      Organophosphorus Pesticides in Man. Perrine, U. S. EPA Office of
      Research and Monitoring Analytical Manual, Sec. 6, 1972.

  5.   Shafik,  T.M., H. C.  Sullivan and H. F. Enos.  A Method for the Deter-
      mination of 1-Napthol in Urine.  Bull, of Environ. Contam. and Toxicol.
      6:34-39,  1971.

  6.   Sherma, J. and T. M. Shafik.  A Multiclass, Multiresidue Analytical Method
      for Determining Pesticide Residues in Air.  Submitted for Publication

  7.   Holmes, J.H.  Clinical Studies of Exposure to the Organophosphorus
      Pesticides.  New York, Academic Press, 1965.

  8.   Brown, H.W.  Electroencephalographic Changes and Disturbance of  Brain
      Function Following Human Organophosphate Exposure.  Northwest Med. 70:
      845-846, 1971.

 9.  Metcalf, D.R.  and J. H.  Holmes.   EEC,  Psychological,  and Neurological
     Alterations in Humans with Organophosphorus Exposure.  Ann. N.Y.  Acad.
     of Sci. 160:357-365, 1969.

10.  Hunter, C.G.,  J.  Robinson,  and M.  Roberts.   Pharmacodynamics of  Dieldrln
      (BEOD)  Part II:  Ingestion by Human Subjects for  18-24 Months and Post-
     Exposure for 8  months.  Arch. Environ. Health 18:12-21,  1969.
                                     40

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EEC


Dasanit

       R
  (R)
                                    Section IX


                                    GLOSSARY


                 electroencephalograph


                 (0,0-diethyl-O-p  (methylsulfinyl) phenyl phosphorothioate ~ fenaulfo-
                                                                               thion
Thimet"        0,0-Diethyl S-(ethylthio)-methyl phosphorodlthioate - phorate


Di-SystonR     0,0-Diethyl S-(2-(ethylthio)ethyl) phosphorodithloate - disulfoton


DMF             0.,.0-Dimethyl phosphate


DBF             £,_0-Diethyl phosphate


DETP            £,j>-Diethyl phosphorothionate  thiophosphoric acid


DMTP            £,£-Dimethyl phosphorothionate thiophosphoric acid


M.D.            Non-detectable


ppm             parts per million


parathion       0,0-Diethyl-O-p-nitrophenyl phosphorothioate


Methyl parathion 0.0-Dimethyl 0-p-nitrophenyl phosphorothioate


parathion 8E    8 pounds Ethyl Parathion in a gallon of liquid


parathion 6-3   Mixture of Ethyl and Methyl parathion


                A mixture of chlorinate camphene compounds of uncertain identity -
                                           *        e  combined chlorine 67-69%)

                (0,0-Dimethyl S-(N-methylacetamide)phosphorodithloate - Timethoate


                2, chloro, 4, ethyl 6 iso proylamlno-S, trlazene


                (0,0-Diethyl S-(p-chlorophenylthio) methyl phosphorodithioate)


                mevinphos (0,0-Diaethyl 2 methoxycarbonvl-1-methvl vinvl  ohosohate)


                manganese-Ethylenebisdithlocarbamate           - maneb


                methomyl. S-Methyl N (methyl-carbamoyl)oxy)thioacetlmldate


                Baellluiq thurineienala (crystalline bacterial toxin)


                dimethyl phosphate of 3-hydroxy-N-methyl-ciscrotonamlde ~ nonocrotoph(


                methamidophos °»Si-Dimethyl phosphoramldothioate


                (1,2 dibromo-2,2-dichloroethyl dimethyl phosphate)- naled
    (R)


(R)


  (R)


   (R)
Toxaphene


Gygon


Aatrex


Trithion


PhoadrinR


DithaneR


LannateR


DipelR


AzodrinR


MonitorR


DibromR


Guthlon R
                0,0-Dimethyl s-(4-oxo-l,2,3-benzotriazine-3-(4-H-yl methyl)- azinpho;

                                                                            methyl
                                     41

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 ml/mOsm/1

 3,5,6-TC pyrldinol

 IPP

 PNP
 Baygon
       (R)
 Dursban
        (R)
 RBC

 PI.

 ChE

 n

 HUD

 2,4, 5-T

 2,4,5-TCP

 2,4-DCP

 Ronnel(R)


 tr

 VC-13


 ug/ml

 CFM

  BHC

tfT BHC
milllliter per milliosmols per liter

3,5,6-trichloro-2 pyridinol

2-lso-propoxyphenol

paranltrophenol

Aprocarb 2-isopropoxyphenly-N-methyl carbamate

chlorpyrifos 0,0-Dimethyl 0-p-nitrophenyl phosphoro-
thioate

dichlorvos  2,2-dlchlorvinyl dimethyl phosphate

red blood cell

plasma

chollnesterase

number of persons

Housing & Urban Development (Dade County)

(2,4,5-tricholorophenoxyacetlc acid)

(2,4,5-trlcholorphenol)

2, 4-dichlorophenol

fenchlorphos 0,0-Dimethyl (2,4,5-tricholorphenol
 phosphorothioate

trace amounts

dichlofenthion (0-(2,4-dichlorophenyl) 0,0-dietyl
phosphorothionate)

microgram per milliliter

cubic feet per minute

alpha isomer of 1,2,3,4,5,6-hexacloro-cylcohexane

Lindane  gamma isomer of 1,2,3,4,5,6-hexachloro-
cyclo-hexane of 99f% purity

cubic meter
                                     42

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 BIBLIOGRAPHIC DATA
 SHEET
1. Report No.
 EPA-650/1-74-009
3. Recipient's Accession No.
4. Tide and Subtitle


     PESTICIDES EPIDEMIOLOGICAL FIELD STUDIES
                                                5- Report Date
                                                  June 1974
                                                                    6.
7. Author(s)
         Dr.  John E. Davies
                                                  Performing Organization Rept.
                                                  No.
9. Performing Organization Name and Address

     University of Miami School  of Medicine
     1600 N.  W.  10th Avenue
     Miami,  Florida 33152
                                                10. Proiect/Task/Work Unit No.
                                                11. Contract/Gram No.
                                                   Grant  No. 802112
                                                Contract  No. 68-02-1145
12. Sponsoring Organization Name and Address
     U. S.  Environmental Protection Agency
     Pesticides and Toxic Substances Effects Laboratory
     National Environmental Research Center
     Research Triangle Park, North Carolina 27711
                                                13. Type of Report & Period
                                                  Covered Annual Report

                                                 12/18/72 - 12/31/73
                                                14.
15. Supplementary Notes
16. AbstracisTne use of the multi-residue techniques for urinary metabolites appeared to  be
provide  the key to worker  exposures from mixed  organophosphate and  carbamate exposures.
The  findings of Shafik et  al.,  were corroborated,  and highly significant inverse correla-
tions  of cholinesterase levels  with the dialkyl phosphate urinary metabolites were demon-
strated  under conditions of  work exposure to  the more toxic organophosphates such as
ethyl-methyl parathion and Phosdrin.  DEP, DETP, DMP and DMTP were  the only alkyl phos-
phates regularly found in  these occupational  exposures.   Following  8  hours of a work  ex-
posure to Dursban the appropriate dialkyl phosphate and  phenolic derivatives persisted
in the urine for 36 hours  and longer.   These  two groups  of metabolites were exceedingly
informative in acute pesticide  poisong and provided etiologic specificity and information
for  the  continued need of  atropine therapy.   Severe and  protracted  illness was observed
following intoxication by  more  polar and fat  soluble organophosphates especially di-ethyl
dichlorophenol phosphorothioate (VC-13).  This  pesticide and its metabolites were detecte
in serum and in urine 48 days after exposure.    Additionally air sampling of the South
Florida  environment confirmed the disappearance of  nr>T and -it-c
17. Key Words and Document Analysis. 17a. Descriptors

       Urinary metabolite  surveillance of the occupational pesticide worker
       Urinary metabolites, di-alkyl phosphate and  phenolic metabolites  in multi-residue
       techniques.
       Fat  soluble organophsophate intoxication - diagnosis and surveillance therapy.
       The  changing profile of pesticide environmental  air sampling.
17b. Identifiers/Open-Ended Terms
17e. COSATI Field/Croup
 18. Availability Scatement
                                    19. Security Class (This
                                       Report)
                                    	UNCLASSIFIED
                                                               iqClASSlFIEE
                                                               jrity Class (Th
                                    20. Security Class (This
                                       Page
                                    	llNCLASSIFIED
          21. No. of Pages
          22. Price
FORM NT IS-35 (IO-7OI
                                             43
                                                                               USCOMM-OC 40329-P7I

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