EPA-660/3-74-025
DECEMBER 1974
                                      Ecological  Research  Series
The  Fate  of Select Pesticides in  the
Aquatic  Environment
                                     National Environmental Research Center
                                       Office of Research and Development
                                       U.S. Environmental Protection Agency
                                              Corvallis,  Oregon 97330

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                      RESEARCH REPORTING SERIES
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          1.   Envi ronmental Health Effects Research
          2.   Environmental Protection Technology
          3.   Ecological Research
          4.   Environmental Monitoring
          5.   Socioeconomic Environmental Studies

This report has been assigned to the ECOLOGICAL RESEARCH STUDIES
series.  This series describes research on the effects of pollution
on humans, plant and animal species, and materials.  Problems
are assessed for their long- and short-term influences.  Investigations
include formation, transport, and pathway studies to determine
the fate of pollutants and their effects.  This work provides
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                              OPTS-TECHNICAL INFORMATION CENTER
                                 EPA-660/3-74-025
                                 December 1974
    THE FATE  OF SELECT PESTICIDES IN THE

              AQUATIC ENVIRONMENT
                     by
              James R. Sanborn
              Project R-800736
           Program Element 1BA023
            ROAP 21 AIM, Task  02
               Project Officer
             Mrs.  Doris F. Paris
Southeast Environmental Research  Laboratory
  National Environmental Research Center
          Athens,  Georgia  30601
   NATIONAL ENVIRONMENTAL RESEARCH CENTER
    OFFICE OF  RESEARCH AND DEVELOPMENT
   U. S. ENVIRONMENTAL PROTECTION  AGENCY
         CORVALLIS, OREGON  97330

  For sale by the Superintendent of Documents, U.S. Government Printing Office
         Washington. D.C. 20402 - Stock No. 5501-0099S

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                               ABSTRACT

In this study 17 organic pesticides and five industrial chemicals were
examined in a terrestrial-aquatic model ecosystem in an effort to
determine their persistence and accumulation by the organisms of this
system.  Several classes of pesticides are represented as one or more
insecticides, herbicides, miticides or plasticizers were investigated
in this system.  The use of this system for examining uptake and per-
sistence of widely used agricultural chemicals provides the necessary
data for comparison of field data to provide a framework which can be
used to assess the potential environmental impact of new pesticides
before they are given a recommendation for generalized use.

The data obtained from this work suggest that this model ecosystem is
useful for the determination of the uptake and persistence of pesti-
cides by the organisms.  In general, it was found that most chemicals,
with the exception of the persistent soil insecticide, dieldrin, under-
went extensive degradation under the experimental conditions of the
system.  Dieldrin was exceptional in its behavior in that > 96% of the
radioactivity isolated from the organisms was unchanged dieldrin,
clearly indicating the extreme inertness of this chlorinated hydrocarbon
to undergo biological or chemical modification.

This report was submitted in fulfillment of Project R-800736 by the
Illinois Natural History Survey and Board of Trustees, University of
Illinois, under the sponsorship of the Environmental Protection Agency.
Work was completed as of June 30, 1973.
                                   ii

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                                CONTENTS
Abstract

List of Tables

Acknowledgements

Section

               Conclusion

               Introduction

               Examination of Select Insecticides

               Examination of Select Herbicides
  I

 II

III

 IV

  V
   VI

  VII

 VIII
Examination" of a Miticide, Select Plasticizers,
  a Fungicide and a Bacteriostat

References

Publications

Chemical Nomenclature for Compounds Examined
  in a Model Ecosystem
                                                                      Page
 11

 iv

vii



  1

  2

  3

 30


 56

 76

 80


 82
                                   iii

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                                 TABLES

Number                                                                 Page
          Concentrations  (ppm) of Bux^ and metabolites
          in organisms in a model ecosystem                              4
   2      Concentrations  (ppm) of Bux^ and metabolites
          in water of a model ecosystem                                  5
                                                14
   3      Concentrations  (ppm) of ring-labelled   C
          carbaryl and metabolites in  organisms and water
          of a model ecosystem                                           7

   4      Concentrations  (ppm) of ring-labeled carbofuran
          and metabolites in organisms of a model ecosystem             10

   5      Concentrations  (ppm) of ring-labeled carbofuran
          and metabolites in water of  a model ecosystem                 12

   6      Concentrations  (ppm) of carbonyl-labeled carbofuran
          and metabolites in organisms of a model ecosystem             13

   7      Concentrations  (ppm) of carbonyl-labeled carbofuran
          and metabolites in water of  a model ecosystem                 15

   8      Concentrations  (ppm) of dieldrin and metabolites
          in organisms of a model ecosystem                             17

   9      Concentrations  (ppm) of dieldrin and metabolites
          in water of a model ecosystem                                 18

   10      Concentrations  (ppm) of lindane and Aroclor 5460^
          and metabolites in organisms in a model ecosystem             21

   11      Concentrations  (ppm) of lindane and Aroclor 5460^
          and metabolites in water in  a model ecosystem                 22
   12      Concentrations  (ppm)  of  Orthene0^  and metabolites
           in organisms  in a model  ecosystem                            25

   13      Concentrations  (ppm)  of  Orthene-^  and metabolites
           in water in a model  ecosystem                                26

   14      Concentrations  (ppm)  of  parathion and metabolites
           in organisms  of a model  ecosystem                            28

   15      Concentrations  (ppm)  of  parathion and metabolites
           in water of a model  ecosystem                                29

   16      Concentrations  (ppm)  of  alachlor and metabolites
           in organisms  in a model  ecosystem                            31
                                    IV

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Number                                                                 Page

  17      Concentrations  (ppm) of alachlor and metabolites
          in water in a model ecosystem                                 32

  18      Concentrations  (ppm) of propachlor and metabolites
          in organisms in a model ecosystem                             33

  19      Concentrations  (ppm) of propachlor and metabolites
          in water in a model ecosystem                                 34

  20      Concentrations  (ppm) of Bladex^ and metabolites
          in organisms in a model ecosystem                             36
  21      Concentrations (ppm) of Bladesc^ and metabolites
          in water in a model ecosystem                                 37

  22      Concentrations (ppm) of Bentazorr-^ and metabolites
          in organisms in a model ecosystem                             39
  23      Concentrations (ppm) of Bentazorr-^ and metabolites
          in water in a model ecosystem                                 40

  24      Concentrations (ppm) of dicamba and metabolites in
          organisms in a model ecosystem                                42

  25      Concentrations (ppm) of dicamba and metabolites in
          water in a model ecosystem                                    43

  26      Concentrations (ppm) of 2,4-D and metabolites in
          organisms in a model ecosystem                                45

  27      Concentrations (ppm) of 2,4-D and metabolites in
          water in a model ecosystem                                    46

  28      Concentrations (ppm) of pyrazon and metabolites
          in organisms in a model ecosystem                             48

  29      Concentrations (ppm) of pyrazon and metabolites
          in water in a model ecosystem                                 49

  30      Concentrations (ppm) of Trifluralirt-^1 and
          metabolites in organisms in a model ecosystem
          after sorghum treatment                                       51

  31      Concentrations (ppm) of Trif luraliir^ and
          metabolites in water in a model ecosystem
          after sorghum treatment                                       53

  32      Concentrations (ppm) of Trifluralirf^ and
          metabolites in organisms in a model ecosystem
          after sand treatment                                          54

  33      Concentrations (ppm) of Trifluralirt^' and
          metabolites iu water in a model ecosystem
          after sand treatment                                          55

                                    v

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Number                                                                 Page
  34      Concentrations  (ppm) of Banomite^ and metabolites
          in organisms in a model ecosystem                            58

  35      Concentrations  (ppm) of Banomite*^and metabolites
          in water in a model ecosystem                                 59

  36      Concentrations  (ppm) of di-n-octyl phthalate and
          metabolites in  organisms of a model ecosystem                 61

  37      Concentrations  (ppm) of di-ri-octyl phthalate and
          metabolites in  organisms of a model ecosystem                 62

  38      Distribution of chlorinated biphenyls and  their
          degradation products in the model ecosystem                   64

  39      Concentrations  (ppm) of Captan and metabolites
          in organisms in a model ecosystem                            68

  40      Concentrations  (ppm) of Captan and metabolites
          in water in a model ecosystem                                 70

  41      Concentrations  (ppm) of hexachlorophene and
          metabolites in  organisms in a model ecosystem                 73

  42      Concentrations  (ppm) of hexachlorophene and
          metabolites in  water in a model ecosystem                     74

  43      Accumulation factors for fish and snail for
          compounds examined in  the model ecosystem                     75
                                   vi

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                          ACKNOWLEDGEMENTS

The author would like to acknowledge several persons during this
investigation.  The initial studies were directed by Dr. G. M. Booth
and carried out by Drs. Ching-Chieh Yu and Dale J. Hansen.  Also, an
acknowledgment is in order for Dr. William F. Childers and Mr. Lowell
Davis who reared the aquatic organisms and provided useful information
regarding the proper procedures for the maintenance of these organisms.
Valuable discussions with Professor Robert L. Metcalf, the developer
of this system, provided important insights regarding the interpreta-
tion of segments of the data in this report.  In addition, the skillful
assistance of M. Kathryn McClendon who helped finish the latter stages
of this work as well as provided assistance during the preparation of
this report is acknowledged.   Finally,  the outstanding laboratory
facilities of the Illinois Natural History Survey and the University
of Illinois are gratefully acknowledged.
                                 vii

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                              SECTION I

                             CONCLUSION

The data presented in this report demonstrate the usefulness of  a
terrestrial-aquatic model ecosystem for the prediction of the persist-
ence and uptake of selected organic chemicals.  Only the soil
insecticide, dieldrin, and the herbicide, TrifluralinB/, were found  to
accumulate over the concentration in the water in the fish and snail.
The experiment with lindane and the extender, AroclouB) 5460, yielded
information that indicated the snail accumulation was unaffected as
compared to lindane examined alone in this system, while the fish
accumulation was increased slightly.  Other chemicals examined in this
system were three pure ^C labeled polychlorinated biphenyls which
accumulated in increasing amounts in the fish and snail as the number
of chlorine substituents was increased.  In addition, investigation of
the fate of the phthalate plasticizer, di-ri-octyl phthalate (DOP) ,
demonstrated substantial accumulation in the fish and snail.  Neither
the bacteriostat, hexachlorophene, the fungicide, captan, nor the
miticide, BanomiteS', accumulated to significant amounts in the fish
or snail.

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                             SECTION II

                            INTRODUCTION

The utilization of a model ecosystem to examine the persistence  and
uptake of pesticides, or any other organic or inorganic  chemical, has
been shown recently to be a valid method for predicting  the  behavior of
these materials in a terrestrial-aquatic environment  (Metcalf  et al.,
1971; Metcalf et_ al., 1973).  Though it has been adequately  described
before, a brief description of the terrestrial-aquatic model ecosystem
will be useful for interpreting the contents of this  report.

The system is housed in a glass aquaria (25 x 30 x 45 cm)  and  contains
a sand-water interface consisting of 15 kg of sterilized white quartz
sand and 7 liters of standard reference water (Freeman,  1953).   Sorghum
(Sorghum halpense) is grown in the sand to a height of 10-12 cm  which
is then treated on the leaves with 5 mg of a radlolabeled  pesticide
dissolved in acetone.  Each compound was run in duplicate  through the
model ecosystem.  The design of this system and treatment  level  corres-
pond to a farm pond surrounded by a watershed under cultivation  that
has been treated 1 Ib/acre (VL.12 kg/hectare).  After the  sorghum has
been treated, saltmarsh caterpillar larvae (Estigmene acrea) are added
to eat the treated plant and therefore simulate both  the first member
of a food chain as well as act as an effective distributing  agent for
the labeled pesticide inside the system.  The water contains several
members of a fresh water aquatic food chain: namely,  snails  (Physa sp.),
water fleas (Daphnia magna) and green filamentous algae  (Oedogonium
cardiacum).

After 27 days mosquito larvae are added to the system to become  another
member of the food chain and 3 days later a mosquito  fish  (Gambusia
affinis) is added to become the final segment of the  system.   At the
end of 33 days the entire system is taken apart and the  organisms and
water are analyzed for radioactivity by extraction with  organic  solvents
and quantitation is carried out by counting the extracts by  liquid
scintillation techniques.  In addition, the extracts  are spotted on
thin-layer  chromatographic plates, developed with appropriate  solvents
and exposed to X-ray film to locate and identify the  chemical  composi-
tion of the solvent extracts.  Identification of metabolites is  made by
co-chromatography with proposed metabolites as well as techniques of
infrared, nuclear magnetic resonance and mass spectrometry.  Once the
identity of the metabolites or degradation products is known,  then
quantitative determination can be made on the propensity of  an organic
chemical or its metabolites to be concentrated from the  water  by the
organisms  of the system.

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                              SECTION III

                   EXAMINATION OF SELECT INSECTICIDES

 The first segment of this report is concerned with the behavior of
 select insecticides  in the model ecosystem.   Initially, the system was
 operated with several additional organisms besides the standard fish,
 snail, mosquito,  Daphnia and algae; but later it was decided to delete
 these additional  organisms so that more valid comparisons could be made
 with data derived by other investigators using the system developed by
 R.  L. Metcalf and co-workers.
 The first  insecticide examined is  the carbamate insecticide, BuxB),
 which is a 3:1 mixture of m-(l-ethylpropyl)phenyl fj-methylcarbamate and
 m-(l-methylbutyl)phenyl IJ-methylcarbamate .   This insecticide has shown
 promise as a soil insecticide for  control of pests under corn.   It is
 moderately toxic to warm-blooded animals  as  it has acute oral and
 chronic U^Q'S of 87 and 400 mg/kg to the rat.  The metabolism of
 m-(l-methylbutyl)phenyl N -methyl carbamate has been examined in rats and
 the primary excretion products found in the  urine are conjugates of the
 phenol, m-(l-methylbutyl)phenyl tl-hydroxymethyl methylcarbamate and
 m-(l-methyl-l-hydroxybutyl)phenyl  N^methylcarbamate (Sutherland et al. ,
 1970).   Further, application of Bux® to the  soil in which maize was
 grown resulted in no carbamate residues in the plants and primary
 metabolites isolated from the soil were the  result of hydrolysis of the
 carbamate  to the phenol and oxidation of  Bux-S' to m-(l-methyl-l-
 hydroxybutyl)phenvl N-methylcarbamate.  From these studies it was cal-
 culated that BuxB'had a soil half-life  between 1-3 weeks (Knaak, 1971).

 Examination of the data in Tables  1 and 2 suggest that Bux3y is  a non-
 persistent insecticide as none of  the snails, mosquitoes or fish
 contain residues of the intact insecticide.   Further, the extractable
 radioactivity from these organisms averaged  18% and was in such small
 quantities that the metabolites were uncharacterizable.  The only animal
 that contained Bussx was the crab (Uca mane lens is)  which had 0.0498 ppm
 and several other unidentified spots on the  chromatogram.   While the
 crab contained Bux-B/ , nevertheless the  extractable radioactivity only
 accounted  for about 15% of the total radioactivity in the organism.

 In contrast to the animals of this system, both the algae with  0.980 ppm
 and E lode a with 0.245 ppm contained considerable amounts of Bux®.
 While these two organisms contained substantial amounts of the  parent
 compound,  the unextractable radioactivity remained high as 81%  and 73%
 in algae and Elodea, respectively, were acetone insoluble materials.

 The water  portion of the ecosystem contained small amounts of Bux® with
 a value of 0.0000953 ppm for the combined total of unhydrolyzed and
 hydrolyzed water.   In addition to  the Bux® there were several unidenti-
i f ied metabolites in the water.   The unextractable  radioactivity for the
 unhydrolyzed -and hydrolyzed water  was slightly higher at 94%.

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                                               Table  1
                       Concentrations  (ppm) of Bux&) and metabolites  in  organisms
                                         in a model ecosystem
      Compound
!£/


II

III

IV

Extractable 1

Unextractable

Grand Total 14C
                    R//
            Algae

    0.98   0.980

0.95-0.62  0.474

0.62-0.28  0.252

0.28-0.02  0.074

    0.00   0.00111

           1.783

           7.825

           9.608
Clamfe/
--
--
--
--
--
0.0206
0.0826
0.103
Crab
0.0498
0.168
0.056
0.0079
0.00508
0.287
1.59
1.877
Daphnia
--
--
--
--
--
0.128
1.42
1.548
Elodea
0.245
0.107
0.206
0.119
0.268
0.945
2.51
3.455
Fish
--
--
--
--
--
0.0449
0.230
0.275
Mosquito
--
--
--
--
--
0.178
0.602
0.780
                                                                                                0.119

                                                                                                0.662

                                                                                                0.781
   Microfiber absorbent sheets impregnated with silica gel,  acetone-n-hexane,15:85 by volume
—' Clam died 7 days after application of Bux^ to system
—I Roman numerals - unknown spots

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     Compound
                                Table 2

             Concentrations (ppm) of Bux® and metabolites
                     in water of a model ecosystem

                        a/
Unhydrolyzed Water
Hydrolyzed Water
Bux® 0.98
b/
I-' 0.95-0.62
II 0.62-0.28
III 0.28-0.02
IV 0 . 00
Extractable ^C
Unextractable ^C
Grand Total 14C
0.0000908

0.0000454
0.00000258
--
0.000000090
0.00014
0.00281
0.00295
0.00000459

0.0000360
0.0000126
0.0000589
0.000103
0.000215
0.00288
0.00311
a/
—   Microfiber absorbent sheets impregnated with silica gel, solvent:
      atetone-n-hexane, 15:85 by volume
_'  Roman numerals - unknown spots

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In summary, it can be concluded from the data obtained  from  the
examination of Bux© in the model ecosystem that this carbamate insecti-
cide does not accumulate in any of the animals of the system, though  it
appears to be concentrated from the water by the algae  and Elodea.
10,283x and 2,571x, respectively.  This phenomenon  is peculiar in that
this substantial accumulation does not appear to cause  any damage to
these plants.  The probable explanation for the absence of BuxS'  in
either the fish or mosquito relates to the water instability of  this
material.  The program of operation of this system  places these  two
organisms in the system on the 27th day for the mosquito and the 30th
day for the fish, and by this time the Bux® has undergone hydrolysis
to the phenol.
Sevii®, or carbaryl, was the first commercial carbamate  insecticide  to
find widespread use in the early 1950's.  Consequently,  because  of its
extensive use, the metabolism of this  insecticide has been  thoroughly
examined by numerous investigators and an adequate review of  the
metabolism and degradation of Sevin® has been outlined  (Fukuto and Sims,
1971).  Briefly, the primary routes  of metabolism are ring  hydroxylation,
hydrolysis to yield QJ-napthol and transformation of  the  N-
methylcarbamoyl group to the N-hydroxymethyl group.

Examination of the data in Table 3 gjLyes clear evidence  that  this car-
bamate  insecticide is similar to Buses' in its metabolic  and  degradative
behavior in this system.  While all  of the  organisms contained substan-
tial radioactivity, none of the organisms contained  carbaryl.  The
quantity of unextractable radioactivity in  the organisms was  on  the
average about 78%, which was slightly  lower than that obtained for BuxSl

The aqueous segment of the ecosystem did not contain any carbaryl, but
two identifiable metabolites in ppt  concentrations were  found; namely,
N-hydroxymethyl carbaryl at 88 ppt and 7-OH carbaryl at  99  ppt.  Other
metabolites were co-chromatographed  with the extract from water, but
the radioactive spots did not correspond with any of the known com-
pounds. Again, as in the case of Bu*D, substantial  amounts of
unextractable radioactivity (72%) were found in the  water.  The  greater
susceptibility of carbaryl to undergo  degradation to the numerous
degradation products isolated from the water perhaps can be ascribed to
the lower aromatic character of the  napthalene ring  which allows facile
hydroxylation to polar, water soluble  metabolites.

                              Carbofuran

The last carbamate investigated in the present study is  carbofuran,  an
outstanding soil insecticide used for  control of insects which affect
corn.   It was of particular importance to examine this  insecticide  in
relation to the state of Illinois as nearly 40% of the  corn in 1972  was
treated with either an organic phosphate or carbamate  (Kuhlman and
Cooley, 1973).  Further, since aldrin, heptachlor and chlordane  were
not recommended for use  in Illinois  to control insects  which  are pests

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Table 3
Concentrations (ppm) of ring-labelled -"-^C carbaryl and metabolites
            in organisms and water of a model ecosystem

    /
                           Clam     Crab   Daphnia  Elodea  Mosquito  Fish


                                   0.118      —    0.057
Compound
I
II
carbaryl
III
A
IV
V
VI
B
C
D
VII
E
VIII
• Rf
0.95
0.87
0.85
0.83
0.79
0.67
0.53
0.47
0.35
0.30
0.26
0.22
0.18
0.12
Water
--
0.000161
--
0.000155
--
0.000221
0.00006
0.000133
0.000081
--
--
0.000018
0.000099
0.000765
Algae
0.175
--
--
--
--
--
--
--
--
--
--
—
--
__
                                                Snail


                                                0.03
      0.0098
                                                    0.085
0.86

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                                                 Table 3 (con't.)
oo
     Compound

IX

X

Extractable ^C

Unextractable 14C

Grand Total ^C
R a/
0.08
0.00



Water
0.00151
0.00748
0.0107
0.0267
0.0374
                                           Algae   Clam
                                           0.614

                                           0.789

                                           3.964

                                           4.753
0.286

1.341

1.627
 Crab   Daphnia  Elodea  Mosquito  Fish   Snail

0.0137     --    0.039

0.257      --    0.909      --     0.091  0.45

0.398    0.295   1.089    0.360    0.091  1.34

0.738    2.385   3.511    2.657    0.337  3.79

1.136    2.681   4.600    3.017    0.428  5.13
      £  Silica Gel GF-254, chloroform-methanol, 49:1 by volume
      —'  Roman numerals - unknown spots
      A  1-napthol
      B  N-hydroxymethyl carbaryl
      C  5-hydroxy carbaryl
      D  4-hydroxy carbaryl
      E  7-hydroxy carbaryl

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of corn (Kuhlman, 1973), it is clear that substitutes must  be  found,
such as carbofuran.  It was therefore paramount that this insecticide
be examined in terms of its persistence and uptake by the organisms  of
this model ecosystem.  An additional factor, which made  it  essential
that this insecticide be examined, is its extreme toxicity  to  warm-
blooded animals as it has an oral LD5Q for the rat of 4 mg/kg  (Metcalf,
1971).

Substantial work has been carried out on the metabolism of  carbofuran
and it has been adequately reviewed (Fukuto and Sims, 1971).   The basic
scheme of metabolism in plants and animals is oxidation of  the 3-carbon
to yield 3-hydroxy and 3-ketocarbofuran.  Additional sites  of  metabolism
involve aromatic hydroxylation and modification of the N-methylcarbamoyl
moiety to give N-hydroxymethyl carbofuran.  Environmental studies in
soil have shown that carbofuran is hydrolyzed to yield the  phenol,
2,3-dihydro-7-hydroxy-2,2-dimethylbenzofuran.  The half-life for the
transformation of the carbamate to the phenol was determined to be
about 30 days.  After the transformation of carbofuran to the  phenol,
the phenol becomes an unextractable residue with a half-life for
extractability of about 7 days (Knaak, 1971).

Examination of the data in Tables 4 and 6 for carbofuran and its degra-
dation products reveals that none of the organisms contained residues
of carbofuran.  In the carbonyl labeled carbofuran there were  several
unknown compounds isolated from Elodea as well as 3-ketocarbofuran
(0.035 ppm), tl-hydroxymethyl carbofuran (0.035 ppm) and 3-
hydroxycarbofuran (0.0118 ppm).  While there were not as many
metabolites isolated from the ring-labeled carbofuran, Elodea  contained
small amounts of carbofuran phenol, 3-hydroxycarbofuran as well as a
small amount of an  unknown material which had an Rf of 0.36.  Again,
as previously observed for BuxB) and Sevir®, most of the radioactivity
for carbofuran was unextractable by acetone as values for ring- and
carbonyl-labeled carbofuran were 69% and 77%, respectively.   These high
figures appear to be characteristic of carbamates in general and
indicate their susceptibility to undergo degradation or metabolism to
polar, uncharacterizable, unextractable species.

In the water portion of the carbonyl-labeled carbofuran ecosystem
(Table 7) small amounts (0.000538 ppm) of the unchanged insecticide were
isolated from the system.  In comparison in the ring-labeled system
(Table 5) the tic resolution was not as precise because carbofuran and
3-ketocarbofuran phenol overlapped.  However, there cannot be more than
0.00128 ppm of carbofuran, if it is assumed that the entire spot is
carbofuran.  Other spots found in identifiable quantities were 3-
ketocarbofuran, N-hydroxymethyl carbofuran, carbofuran phenol  and
3-hydroxy carbofuran.  None of these compounds was found in excess of
10 ppt which indicates conclusively that carbofuran degrades to polar,
water soluble materials which do not persist in the water segment of
the ecosystem or remain at high levels.

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                            Table 4

Concentrations (ppm)  of ring-labeled carbofuran and metabolites
               in organisms of a model ecosystem
Compound
iV
A
B and carbofuran
C
D
E
F
II
III
IV
V
VI
Extractable ^C
Unextr actable ^C
Grand Total ^C
Rf-'
0.98
0.83
0.76
0.70
0.60
0.53
0.46
0.36
0.28
0.13
0.06
0.00



Algae Clam Daphnia Elodea Fish Frog
0.0462 0.197
0.0130 -- -- 0.000304
_-
--
-_
--
0.00526
0.000828
0.191
--
--
0.883 -- -- 0.0216 0.305
0.815 1.087 1.089 2.697 0.0725 0.502
4.648 0.368 4.690 2.993 0.413 1.034
5.463 1.455 5.779 5.689 0.485 1.536
Mosquito Snail
0.418 0.567
0.377
__
-_
__
--
-_
-_
-_
._
._
0.552 0.890
1.071 1.645
4.835 6.270
5.906 7.915

-------
                                           Table 4 (con't.)

—' Microfiber absorbent sheets impregnated with silica gel, solvent system: acetone-n-hexane,
     15:85 by volume
b_' Roman numerals - unknown spots
A  Carbofuran phenol
B  Carbofuran and 3-ketocarbofuran phenol
C  3-ketocarbofuran
D  3-hydroxy carbofuran phenol
E  N-hydroxymethyl carbofuran
F  3-hydroxy carbofuran

-------
                                Table 5

    Concentrations (ppm) of ring-labeled carbofuran and metabolites
                     in water of a model ecosystem
      Compound
Unhydrolyzed Water
Hydrolyzed Water
B and carbofuran

C

D

E

F

II

III

IV

V

VI
            14
Extractable   C
Unextractable
              14,
            14
Grand Total   C
0.98
0.83
0.76
0.70
0.60
0.53
0.46
0.36
0.28
0.13
0.06
0.00



0.000267
0.00884
0.000909
0.000136
0.0000758
0.000121
0.000196
0.000537
0.000537
0.000137
0.000230
0.000493
0.0037
0.111
0.115
0.000762
0.00287
0.000375
0.000423
0.000280
0.000242
0.000415
0.00255
0.00143
0.00259
0.00308
0.0166
0.0316
0.0652
0.097
£/ Microfiber absorbent sheets impregnated with silica gel, solvent
     system: acetone-n-hexane, 15:85 by volume
b_/ Roman numerals - unknown spots
A  Carbofuran phenol
B  Carbofuran and 3-ketocarbofuran phenol
C  3-ketocarbofuran
D  3-hydroxy carbofuran phenol
E  N-hydroxymethyl carbofuran
F  3-hydroxy carbofuran
                                  12

-------
                                           Table 6
Compound
             Concentrations (ppm)  of carbonyl -labeled carbofuran and metabolites
                              in organisms  of a model ecosystem
Algae
Clam
Crab    Daphnia   Elodea   Fish   Mosquito  Snail
!>/
II
III
carbofuran
A
B
IV
C
V
VI
Extractable ^C
Unextractable 14-C
Grand Total 14C
0.96
0.86
0.75-0.71
0.73
0.63
0.55
0.51
0.48
0.26-0.00
0.00



0.121
—
__
__
0.064
-_
__
0.038
--
0.0482
0.963 0.02065 0.271
3.000 0.100 0.254
3.963 0.12065 0.525
0.0492
0.1261
0.0309
--
0.0350
0.0364
0.0059
0.0118
0.5025
0.342
0.191 1.145 0.0583
1.092 4.835 0.278
1.283 5.980 0.3363
0.221 0.648
__
__
__
__
__
--
__
--
0.088 0.325
0.435 0.972
1.198 6.518
1.583 7.490

-------
                                           Table 6 (con't.)

£/ Microfiber absorbent sheets Impregnated with silica gel, solvent: acetone-n-hexane,  15:85  by
     volume
_' Roman numerals - unknown spots
A  3-ketocarbofuran
B  N-hydroxymethyl carbofuran
C  3-hydroxy carbofuran

-------
                                Table  7

       Concentrations  (ppm) of carbonyl-labeled  carbofuran and
            metabolites in   water   of  a model  ecosystem
     Compound
Unhydrolyzed Water
Hydrolyzed Water
i/
II
III
carbofuran
A
B
IV
C
V
VI
Extractable l^C
Unextractable 14
Grand Total ^C
0.96
0.86
0.75-0.71
0.73
0.63
0.55
0.51
0.48
0.26-0.00
0.00

C

0.00000491
0.0000136
--
0.000364
0.00000859
0.00000747
--
0.00000128
0.00000464
0.0000103
0.000415
0.00374
0.00416
0.0000070
0.000110
--
0.000174
0.000650
0.0000246
--
0.0000246
0.000073
0.000756
0.00178
0.00259
0.00437
—' Microfiber absorbent sheets impregnated with silica gel, solvent:
     acetone-n-hexane, 15:85 by volume
—' Roman numerals - unknown spots
A  3-ketocarbofuran
B  N-hydroxymethyl carbofuran
C  3-hydroxy carbofuran
                                  15

-------
At this point it may be suitable to make some general comments about
the previous three compounds and their behavior  in  this  terrestrial-
aquatic model ecosystem.  All three were carbamate  insecticides and
were shown to be nonpersistent and did not accumulate in any  of the
organisms of this system.  This property may be  characteristic of all
aryl carbamate insecticides as they have sites for  oxidative  metabolism,
namely, the ring, the IJ-methyl group and aliphatic  side  chains.  In
addition to these sites, which are susceptible to oxidative metabolism,
the carbamoyl group can undergo hydrolysis to a  phenol which  detoxifies
the carbamate and provides a moiety which can be conjugated with sugars ,
phosphate or sulfate.  If the data obtained for  these carbamates in
this model ecosystem is representative of the behavior of aryl N-methyl
carbamate insecticides, then it would appear that the use of  these
insecticides will not present ecological problems related to  persist-
ence and food chain accumulation.

                              Dieldrin

The next pesticide to be discussed is the chlorinated hydrocarbon,
dieldrin.  The unoxidized precursor to dieldrin, aldrin, was  introduced
about 20 years ago as an effective insecticide for  the control of soil
insects, particularly those associated with corn.   The continual use of
this material for over 20 years has resulted in  resistance to this
insecticide and, therefore, the aldrin/dieldrin  treatment of  corn
insects is not particularly effective for some corn pests (Sechriest
and Sherrod, 1973).  The increased emphasis on environmental  quality
has also suggested the discontinued use of aldrin and/or dieldrin as
dieldrin, the oxidation product of aldrin, is very  persistent with a
soil half-life of about 3 years (Wingo, 1968; Hurtig, 1972).  The
persistence is related to the extreme inertness  toward chemical or
biological modification.  For example, metabolism studies with dieldrin
and wheat (Saha, 1970) indicated that less than  37,  of the application
of dieldrin was  transformed into metabolites.  This corroborates
earlier work done with corn, alfalfa and orchard grass that dieldrin
does not undergo substantial transformation by these plants  (Wheeler
e^al. , 1967).

The  inert nature of dieldrin to undergo degradation in plants is con-
trasted by metabolic susceptibility when administered orally  to a
variety of mammals.  For example,  70% of the oral dose to rats is
eliminated, principally via the feces which points  to biliary excretion
 (Heath and Vandekar, 1964).  More  recently, the  structure of  the
principal metabolite from an oral  dose to rats has  been  identified as
6,7-trans-dihydroxydihydro aldrin  with a specific rotation of
     °  +  13.7°.
 The data in Tables  8 and 9  for dieldrin in the model ecosystem demon-
 strate the inert nature of  dieldrin to undergo biological  or  chemical
 degradation.  There was dieldrin in every organism in the  ecosystem
 ranging from a low  of 0.495 ppm in the crab (Uca man i lens is)  to 230 ppm
 in the snail.  Even more important is  that dieldrin constituted nearly
                                  16

-------
                                                Table 8

                           Concentrations (ppm) of dieldrin and metabolites
                                   in organisms of a model ecosystem
Compound
I*>
dieldrin
II
A
B
VII
Extractable C
Unextractable ^C
Grand Total 14C
R
0
0
0
0
0
0



f*
.65
.58
.43
.38
.31
.00



Algae
--
14.96
--
0.20
--
--
15.16
1.23
16.39
Clam
--
2.03
--
--
--
--
2.03
0.028
2.06
Crab
--
0.495
--
--
0.043
--
0.536
0.177
0.715
Daphnia
--
5.07
--
--
--
0.07
5.14
0.10
5.24
Elodea Mosquito
0
2



0
2
0
2
.23
.56
__
__
__
.03
.82 1.35
.14 0.25
.96 1.60
Fish
-
12
-
0
0
0
12
0
13
-
.29
-
.19
.07
.03
.57
.65
.22
Snail
0.866
229.87
0.456
1.11
--
0.044
232.3
1.78
234.1
$.  Silica Gel GF-254, ether-n-hexane, 3:2 by volume
—' Roman numerals - unknown spots
A  9-hydroxy dieldrin
B  9-keto dieldrin

-------
                                Table 9

           Concentrations (ppm) of dieldrin and metabolites
                     in water of a model ecosystem
      Compound
dieldrin

A

B

m^

IV

V

VI

VII

Extractable
14.
Unextractable
              14.
Grand Total
            14,
Rf~
0.58
0.38
0.31
0.18
0,12
0.07
0.04
0.00



Unhydrolyzed Water
0.0020
0.00046
0.00040
0.00034
0.00013
0.00012
0.00093
0.00023
0.0046
0.0028
0.0074
Hydro ly zed Water
--
—
—
--
0.00012
0.00023
0.00108
0.00134
0.0028
--
_ H
a/ Silica Gel GF-254, ether-n-hexane,  3:2 by volume
b_/ Roman numerals  - unknown  spots
A  9-hydroxy dieldrin
B  9-keto dieldrin
                                    18

-------
887o of the total radioactivity  isolated  from  the  organisms.   This
figure includes both extractable and unextractable  radioactivity.
Clearly, dieldrin is not metabolized to  polar, water  soluble molecules
by the organisms of the system, nor is it degraded  by physical or
chemical processes as  little of the radioactivity isolated from the
organisms  (^9%) is unextractable.  This  figure contrasts  significantly
with that observed for the three carbamate  insecticides  (o<65-75%) or
the phosphates  (^60-70%) which will be reviewed next.   In view of the
low, mixed-function oxidase levels of Gambusia affinis  (Krieger and Lee,
1972) and Physa (Metcalf ejt al., 1971),  it  is not surprising that
dieldrin is accumulated by these organisms  as they  appear to be inca-
pable of transforming  the lipid soluble molecule, dieldrin,  into water
soluble metabolites.

The distribution of degradation products of dieldrin  in the  water
(Table 9) differs slightly from the distribution  in the organisms as
dieldrin constitutes only 25-28% of the extractable radioactivity.
Furthermore, dieldrin  is in lower concentration (0.0020 ppm)  than was
isolated from the organisms.  The reduced amount  of dieldrin in the
water, as compared with organisms, is probably related to the low water
solubility, 0.25 ppm (Gunther et_ al., 1968).  If  the  amount  of dieldrin
isolated from the water is divided into the various concentrations  of
dieldrin observed in the organisms, then the tendency  for dieldrin to
accumulate in the various organisms can be calculated.  The concentra-
tion factors for the snail, algae and fish  are approximately 115,000,
7,500 and 6,100, respectively.  These values compare  satisfactorily
with those obtained in the same ecosystem experiment with dieldrin
where the factors were determined to be 61,657 and  2,700  for snail  and
fish, respectively (Metcalf et al., 1973).  More  importantly,  the order
of magnitude for accumulation of dieldrin by aquatic  organisms  has  been
observed to be in the same order of magnitude, particularly  the fish of
the model ecosystem, as that in fish collected from farm  ponds  in
Illinois.  For example, it has been observed that dieldrin is  concen-
trated by several species of fish from 5,000x to  25,000x  the
concentration in the water (Childers and Bruce, 1973).  This  would  again
appear to validate the usefulness of the model ecosystem  to  predict  the
behavior of pesticides in the aquatic environment.

                 Lindane and Aroclor 5460®  (i:5 w/w)

The types and variations of experiments that can be carried  out  in  this
terrestrial-aquatic ecosystem are only limited by the imagination of
the investigator.   In the experiment with lindane and this polychlorin-
ated terphenyl, the intended purpose was to examine the effect  that  this
adjuvant might have on the persistence and  fate of  lindane in the model
ecosystem.   This type of experiment is not without  precedence as  it has
been previously reported that persistence and insecticidal residues of
lindane could be improved by the addition of Aroclor 5460® (Horstein
£t al., 1953;  Lichtenstein, 1969).   It was  concluded that residues of
lindane and Aroclor 5460ty were more persistent because of  the retarda-
tion of the evaporation rate of lindane from treated surfaces
                                                           v

                                 19

-------
(Tsao, 1953).  Further, impetus for conducting this type of paired
interaction experiment is given by the dearth of  information under
controlled environmental conditions of studies involving interactions
of chemicals and the effects that an adjuvant, such as Aroclor 5460^4
may have on the environmental persistence of a pesticide.

The fate of lindane alone in this model ecosystem has been investigated
which makes it possible to compare the effect of  this plasticizer on
the persistence of lindane (Metcalf et_ al. , 1973).  Further, the metabo-
lism of lindane in plants and animals has been adequately reviewed
(Fukuto and Sims, 1971).  The initial transformation which results  in
inactivation of lindane is a dehydrohalogenation  to yield pentachloro-
cyclohexene.  This unsaturated chlorinated cyclohexene undergoes further
transformations to form conjugates with glutathione and dechlorination
to form several isomeric chlorinated phenols.

Examination of the data in Table 10 for the distribution of lindane in
the organisms of the model ecosystem shows that lindane is present  in
all organisms ranging  from about 2.20 ppm for Daphnia to about 26.40 ppm
for the fish.  The value for the fish is about 26x greater than the
value found by Metcalf et al. (1973), which would indicate that the
addition of the terphenyl increases the uptake of lindane by the fish.
However, the variability in concentrations of lindane in the two fish
from the duplicate experiments was unusually high, 48.98 ppm and
3.78 ppm, and therefore, to make an absolute statement about the
accumulation of lindane to be about 28x greater in the presence of
Aroclor 5460® would be tenuous.  It can be stated that there was a
subtle effect of the plasticizer as identifiable  amounts of lindane
were found in the present experiment in the algae, 6.18 ppm; Daphnia,
2.17 i>pm; snail, 5.66  ppm and mosquito, 4.21 ppm. Without the Aroclor
5460JV, no lindane was  found in the algae  or mosquito, but substantial
amounts were  found in  the snail, 0.762 ppm, and fish, 0.975 ppm  (Metcalf
et al., 19731.  Clearly, this preliminary experiment with lindane and
Aroclor 5460») demonstrates the potentiality for studying the interactions
of compounds.  In a recent investigation  it has been demonstrated that
piperonyl butoxide (pb), an insecticide synergist, will increase both
the total radioactivity in the snail and  fish as  well as the concentra-
tion of methoxychlor in these two organisms (Metcalf, 1973).

Other data of interest in the metabolite  distribution from the organisms
is that neither of the two chlorinated phenols, 2,4,6-trichlorophenol
or 2,4,5-trichlorophenol, were found in the organisms, while small
amounts were  found in  the water which will be discussed next.  These
two chlorinated phenols have been reported as urinary excretion products
when rats were fed lindane  (Grover and Sims, 1965).  A final point  that
should be made is the  low percentage of unextractable radioactivity
 (average 7%)  in the organisms.  This figure is similar to the value for
dieldrin and  about one-tenth of the amount obtained for the carbamate
and phosphate insecticides.  The data for the previous experiment with
 lindane in  this model  ecosystem also indicate a low amount of unextract-
able radioactivity, which would seem to imply that the addition of  the
                                  20

-------
                                              Table 10
                  Concentrations (ppm) of lindane and  Aroclor 54603$ and metabolites
                                   in organisms in a model ecosystem
Compound
•A
1 indane
B
C
I*/
II
Extractable ^C
Unextractable ^C
Grand Total 14C
Rf^7
0.75
0.40
0.19
0.13
0.06
0.00



Algae
--
6.178
--
--
--
0.285
6.463
0.803
7.266
Daphnia
--
2.166
--
--
--
0.127
2.293
0.164
2.457
Fish
0.025
26.379
--
--
--
1.004
27.408
0.087
27.495
Mosquito
--
4.212
--
--
--
0.108
4.320
0.309
4.629
Snail
--
5.658
--
--
0.110
0.410
6.178
0.522
6.700
£/ Silica Gel GF-254, petroleum ether-carbon tetrachloride,1:1 by volume
k_/ Roman numerals - unknown spots
A  pentachlorocyclohexene
B  2,4,6-trichlorophenol
C  2,4,5-trichlorophenol

-------
                              Table 11

          Concentrations  (ppm) of lindane and  Aroclor 5460*'
             and metabolites in water in a model ecosystem
     Compound
R
                        i/

Unhydrolyzed Water
Hydrolyzed Water
A
lindane
B
C
&
II
Extractable ^C
Unextractable ^C
Grand Total ^C
0.75
0.40
0.19
0.13
0.06
0.00



0.0000608
0.0125
0.000471
0.000418
0.000433
0.00124
0.01520
0.00740
0.0226
--
—
0.000056
0.000182
0.000352
0.00144
0.00202
0.00430
0.00632
2.1 Silica Gel GF-254, pet ether-carbon tetrachloride, 1:1 by volume
—' Roman numerals - unknown spots
A  pentachlorocyclohexene
B  2,4,6-trichlorophenol
C  2,4,5-trichlorophenol
                                 22

-------
polychlorinated  terphenyl  does  not  affect the metabolism of lindane to
polar, unextractable metabolites.

The distribution of metabolites in  the  water (Table 11)  for this experi-
ment  of  lindane  and Aroclor  5460© is  somewhat different  than if lindane
'^Ls put through the system  alone.  The most striking difference is the
greater  quantity of lindane,  0.0126 ppm,  found in the present experiment
as compared  to 0.00167  ppm (Metcalf et  a±.,  1973),  which is an increase
of about 7.5x.   Further, in  the present experiment  it was possible to
identify small amounts  of  2,4,6-trichlorophenol,  0.00053 ppm and
2,4,5-trichlorophenol,  0.0006 ppm.  If  the concentration of lindane
isolated from the water is divided  into the  fish  and snail, concentra-
tion  factors are derived of  ^jlOOx and 448x, respectively.  The value
obtained for the fish is about  4x that  obtained for the  fish for
lindane  alone (Metcalf  et  al.,  1973), while  the snail value in the
present  experiment is nearly identical.  The interaction of pesticides
and other  chemicals, such  as  a  polychlorinated terphenyl, give results
that  are complex and difficult  to interpret.  Finally, the unextractable
radioactivity from the  water  for the  lindane experiment  alone was about
2.5%  (Metcalf et^ a^., 1973),  while  the  unextractable radioactivity from
the present experiment  amounted to  26%.  The effect of Aroclor 54605' on
the metabolism,  degradation  and persistence  of lindane in the water is
a 10-fold  increase in unextractable radioactivity.

                                Ortheneffi

OrtheneB) is a new organophosphate insecticide which shows moderate
persistence and  a residual activity of  5-10  days.   In field trials con-
ducted from  1968 to 1972 this insecticide was effective  in controlling
pest  species of  aphids  and lepidopterous  larvae.   In contrast to
parathion, this  phosphoramidothioate  insecticide  shows a low order of
toxicity to higher animals as the LD5Q  to male rats is about 945 mg/kg,
which is about lOOx less toxic  than parathion.  In  addition, the 96-hr
TLijQ  for Orthene® is greater  than 1,000 ppm  for trout, 1,725 ppm for
large-mouth bass, 2,050 ppm  for blue  gill, 2,230  for channel catfish,
6,650 for mosquito fish (Gambusia)  and  9,550 ppm  for goldfish.   The
preceding data,  which was  taken from  the  technical  data  sheet available
from  the Chevron Chemical  Company (1972)  indicates  that  this material
is safe  in terms of acute  toxicity  to both fish and mammals.

The metabolism pathways of this insecticide have not been published, but
a number of possibilities  for potential metabolites and/or degradation
products can be  hypothesized.   The  loss of the N-acetyl  group to give
0,j>-dimethyl phosphoramidothioate,  another Chevron  Chemical Company
product, Monitoifiy, is an obvious metabolite  as  similar transformations
are known  to occur from metabolism  studies on N-derivatized carbamates
(Miskus  et^ al.,  1969).  This  transformation  to Monitor® results  in a
more  toxic molecule to  the rat  as the LD5Q of this  compound is  18.9
mg/kg (Chevron Chemical Company, 1972).   Another  transformation  that
OrtheneS) might undergo  is  cleavage  of either the j5-methyl or 0-methyl
moieties to yield the j>-methyl  acetamidophosphorothioic  acid or
                                 23

-------
0_-methyl acetamidophosphoric acid, respectively.  The  cleavage  of
either S^-methyl or ()-methyl could occur after  loss  of  the  acetyl  moiety
from nitrogen to yield either 0-methyl phosphoramidic  acid or j>-methyl
phosphoramidothioic acid.  One  final  transformation that could  occur  is
the removal of the amino group  after  loss  of the  acetyl group to  yield
0_,^-dimethyl phosphorothioic acid.

Examination of the data in Tables 12  and  13 for the organisms and water,
respectively, shows that none of the  chromatographed metabolites  were
found in identifiable quantities in the organisms.   An unusual
occurrence was the formation of a degradation  product  which was less
polar than Orthene®, which had  an Rf  of 0.70,  or  Monitoi®, which  had  an
Rf of 0.79.  This metabolite varied in concentration from  about 0.26
ppm in Daphnia to greater than  2.0 ppm in the  crab.  Since the  site of
the label was ^C j>-methyl, perhaps this  nonpolar species  is not  related
to OrtheneB), but has been utilized as a source of carbon in a structural
entity in the organism from which it  was  isolated.

Finally, as was previously indicated  for  the carbamate insecticide, the
unextractable radioactivity in  the organisms for  Orthene® was high at
about 72%.  This indicates that this  insecticide  breaks down over the
time period of the model ecosystem to nonlipid—partitioning products.
The only product which appears  in the organisms is  nonpolar (Rf,  0.93)
and does not appear to be deleterious.  It is  not surprising to find
that Orthene  does not accumulate in  the  organisms  of this system as
the water solubility of this insecticide  is reported to be about  650,000
ppm (Chevron Chemical Company,  1972).

In the water of this model ecosystem  (Table  13) were small amounts of
Orthene®, 0.000282 ppm, and as  well as small quantities  (0.000478 ppm)
of the unknown which was nonpolar and had an Rf value of 0.93.  This
nonpolar species was accumulated from the water by  the various  organ-
isms from 538x for Daphnia to about 4,300x for the  crab.   Trace amounts
of Monitoisy, 0.000124 ppm, and  the ammonium salt  of JJ-methyl acetyl-
phosphoramidothioate, 0.000015  ppm, were  found in the hydrolyzed  water.
Finally, the high value found for the unextractable radioactivity in
unhydrolyzed water, >99%,and hydrolyzed water, ^97%, indicates  the
susceptibility of Orthenely to undergo degradation to metabolites  of
high water  solubility.

                                Parathion

Parathion is a member of the first generation  of  organophosphorus
insecticides used  in the United States.   It was originally discovered
in Germany  by Gerhard Schrader  during World War II  and since then has
been used for the  control  of insects  which are pests of many crops.
The large-scale use of parathion  is  indicated  by  the 15,259,000 pounds
manufactured  in the United States  in  1970. While there does not  seem
 to be environmental problems associated with the  use of parathion,
nevertheless  it  is paramount  that  insecticides that are highly  toxic  to
warm-blooded  animals  (oral 11)50, rats 4-13 mg/kg) be .examined  to
                                  24

-------
                                                     Table  12
Ol
                            Concentrations  (ppm)  of OrtheneS' and metabolites in organisms
                                                in a model ecosystem
Compound
£>
A
Orthen<®
B
C
II
III
IV
Extractable 14C
Unextractable ^C
Grand Total 14C
Rf-' Algae Clam Crab Daphnia Elodea Fish Mosquito Snail
0.93 0.936 -- 2.038 0.257 0.408 -- 0.797 0.796
0.79
0.70
0.45
0.33
0.25 0.0538
0.11 0.0142 -- -- -- 0.00280
0.00 0.0395 -- 0.247 0.143 0.0098 -- 0.0247 0.130
1.043 0.100 2.284 0.400 0.421 0.0309 0.822 0.927
3.517 0.148 3.631 1.979 1.435 0.0621 2.466 2.769
4.560 0.248 5.915 2.379 1.856 0.0930 3.288 3.696
      —  Silica Gel GF-254,  aluminum plate in 15% acetic acid in benzene-propanol,  1:1 by volume
      —'  Roman numerals  - unknown spots
      A  0,jS-dimethyl phosphoramidothioate
      B  0,^-dimethyl phosphorothioate sodium salt
      C  B_-methyl IJ-acetylphosphoramidothioate ammonium salt

-------
                              Table 13

           Concentrations (ppm) of OrtheneS' and metabolites
                     in water in a model ecosystem
     Compound
rW
A

Orthene®

B

C

II

III

IV

Extractable
14,
Unextractable

Grand Total
                   Unhydrolyzed Water     Hydrolyzed Water
0.93 0.0000617
0.79
0.70 0.0000484
0.45
0.33
0.25
0.11
0.00
0.00011
0.0376
0.0377
0.000416
0.000124
0.000234
--
0.0000150
--
--
0.0000255
0.00082
0.0236
0.0244
—'  Silica Gel  GF-254,  aluminum plate in 15% acetic acid in benzene-
      propanol, 1:1  by  volume
]>/  Roman numerals  - unknown spots
A  (),j>-dimethyl phosphoramidothioate
B  0,j>-dimethyl phosphorothioate sodium salt
C  ^-methyl  II-acetylphosphoramidothioate
                                 26

-------
determine their persistence in the model ecosystem.   Further,  the
investigation of pesticides in this model ecosystem,  such as  parathion
which has had widespread use without apparent accumulation in food
chains, is necessary so that field and model ecosystem data can be
compared to test the validity of the model ecosystem  as a screening
     for pesticide persistence.
The metabolism of parathion has been thoroughly studied  in  animals  and
somewhat less detailed studies have been carried out  in  plants  and  soil.
Briefly, the major alterations of parathion are oxidation to  paraoxon
and cleavage of parathion to diethylphosphorothioic acid and  _p_-
nitrophenol.  Paraoxon can undergo ()-dealkylation to ^-ethyl,
0-jD-nitrophenyl phosphoric acid as well as cleavage to diethyl  phos-
phoric acid and _p_-nitrophenol.

The model ecosystem study with parathion demonstrates the rapid break-
down of this insecticide as the data in Tables 14 and 15 show that  this
phosphate does not persist to any great extent nor does  it  accumulate
in the fish as was observed for the chlorinated hydrocarbon,  dieldrin.
Only the fish (0.100 ppm) contained parathion as all  the other  organisms
had neither parathion, paraoxon nor j>-nitrophenol.  Therefore, it was
 Possible to establish the fate of j>-nitrophenol since the site  of the
 4-C label was in the 2,6-positions of this moiety.  It would  seem
important to establish the fate of j>-nitrophenol and other  phenols  in a
model ecosystem as they are significant constituents of a number of
organophosphorus insecticides.  Again as previously discussed for the
phosphate insecticide, Orthene®, and the three carbamates in  this
report, substantial amounts of the radioactivity, with the  exception  of
the fish, was unextractable (average 65%) .  An explanation  for  the
relatively high amounts of extractable radioactivity  (81%)  in the fish
is the low microsomal activity in the liver of Gambus ia  (Krieger and
Lee, 1973)  which does not metabolize parathion to polar, unextractable
products.

The water segment (Table 15)  of the ecosystems contains  small amounts
of parathion, 0.0003 ppm; paraoxon, 0.00047 ppm; and j>-nitrophenol,
0.00095 ppm.  In addition, there were small amounts of unknown metabo-
lites which had Rf values of 0.97, 0.73, 0.33, 0.09, 0.13 and 0.00.
The accumulation of parathion by the fish over the concentration in the
water was about 335x, which is about 0.054 the value observed for
dieldrin in this model ecosystem.  Probably, the higher water solubility
of parathion, 25 ppm (Gunther et^ al^. , 1968), as compared to dieldrin,
0.25 ppm (Gunther a£ aJ. , 1968), and the greater susceptibility of
parathion to undergo degradation account for the substantially less
concentration of parathion by the fish in this model ecosystem.   The
portion of unextractable radioactivity in the water was smaller than
for Orthene®, as it averaged about 40%.  Perhaps this decrease  from
approximately 98-99% for Orthene® indicates that parathion,  as  compared
to Orthene®, is less degradable and therefore more persistent in the
aqueous compartment of this model ecosystem.
                                 27

-------
N3
00
                                                     Table  14

                          Concentrations (ppm) of parathion and metabolites  in organisms
                                               of a model ecosystem
Compound
&
parathion
A
III
VI
Extractable ^C
Unextractable ^C
Grand Total 14C
Rf-
0.97
0.90
0.55
0.33
0.00



Algae
0.0356
--
--
--
0.3613
0.3969
2.6284
3.0253
Daphnia
--
--
--
--
0.2987
0.2987
0.3126
0.6113
Fish
--
0.1006
0.0086
0.0222
0.0621
0.1935
0.2055
0.3990
Mosquito
--
--
--
--
0.2031
0.2031
0.4685
0.6716
Snail
--
--
--
--
0.2701
0.2701
0.5818
0.8518
      a/
      -  Silica Gel GF-254, diethyl ether-n-hexane, 7:3 by volume
      SL' Roman numerals - unknown spots
      A  j3-nitrophenol

-------
                               Table  15

          Concentrations  (ppm) of  parathion and metabolites
                    in water of a  model  ecosystem
Compound
i
parathion
II
P.-NC-2 phenol
III
paraoxon
IV
V
VI
Extractable 14C
Unextractable ^C
Grand Total 14C
Rf*.'
0.97
0.90
0.73
0.55
0.33
0.25
0.13
0.09
0.00



Unhydrolyzed Water
0.00020
0.00030
0.00006
0.00074
0.00018
0.00031
0.00049
0.00151
0.00222
0.00661
0.0144
0.0210
Hydrolyzed Water
--
--
--
0.00062
0.00007
0.00016
--
0.00123
0.00377
0.00585
0.00853
0.01438
^•'l Silica Gel GF-254, ether-hexane, 7:3 by volume
—  Roman numerals - unknown spots
                                 29

-------
                             SECTION  IV

                  EXAMINATION OF SELECT HERBICIDES

The second aspect of this report is concerned with the  fate of  several
herbicides in this model ecosystem.  While the acute  toxicity of most
herbicides to mammals is low, it still is necessary to  derive informa-
tion which can be used as a predictive measure of the potential for
these pest chemicals to accumulate and persist in the environment.
Therefore, the next section should provide information  about several
members of this class of pesticide chemicals.  It is hoped that
additional herbicides can be examined in this system, particularly
those which currently have a widespread use, so  that a  baseline can be
established for the comparison of new herbicides.

                       Alachlor and Propachlor

Since alachlor and propachlor have similar structures chemically,
namely, 2-chloroacetanlides, the discussion of these two compounds and
their behavior in the model ecosystem will be combined.   Both compounds
are safe  in terms of acute toxicity as the oral  H>50 values for
alachlor  and propachlor are 1,200 mg/kg and  1,500 mg/kg,  respectively.
The soil  persistence of these two herbicides is  1-2 months which is
quite short.  The principal use has been in  the  control of weeds in
corn as preemergence, preplant or postemergence  treatments.  The mode
of action of propachlor in susceptible plants appears to be the inhibi-
tion of utilization of proteinacous and lipid reserves  which are
necessary for plant growth (Dhillion, 1971).

Some work on the metabolism of these  herbicides  in soil and plants has
been carried out, but it is by no means extensive.  Laboratory  studies
in soil indicates the primary degradative route  for alachlor is loss of
the -CH2OCH3 moiety  from nitrogen  (Hargrove  and  Merkle,  1971).  This
transformation is believed to take place on  the  acidic  soil surface.
Higher relative humidity decreases the herbicide contact with the soil
surface which then decreases the rate of this transformation.   Metabo-
lism of propachlor by corn, sorghum and sugarcane plants yields a
conjugate of glutathione as a primary metabolite (Lamoreaux and Tanaka,
1971).

The data  for these two herbicides  for the organisms contained in Tables
16 to  19  reveal conclusively the lack of uptake  of either of these two
herbicides by any of the organisms.   The water had numerous unidentified
metabolites which were not in the organisms  as well as  small amounts of
the parent compounds, 0.0564 ppb for  propachlor  and about 1.05  ppb for
alachlor. The 2-chloro group of the  acetanilde  is a  labile moiety and
could  undergo displacement to yield a substantially more water  soluble
moiety, though no metabolite was available for co-chromatography.  In
addition, as evidenced by  the approximately  17 unidentified compounds,
it can be stated that both of these herbicides are susceptible  to
degradation to water-soluble, nonlipid-partitioning organic compounds.
                                 30

-------
                                              Table 16
     Compound
                     Concentrations (ppm)  of alachlor and metabolites  in organisms
                                         in a model ecosystem
Algae
Crab
&
alachlor
II
III
IV
V
VI
VII
VIII
IX
Extractable ^C
Unextractable ^C
Grand Total ^C
0.70
0.61
0.51
0.43
0.33
0.27
0.20
0.13
0.07
0.00



__
__
-_
--
.-
__
__
__
__
-_
0.0898 0.321
0.569 0.524
0.658 0.845
Daphnia
                                                     0.000

                                                     0.422

                                                     0.422
-' Silica Gel GF-254, methanol-benzene, 5:95 by volume
—' Roman numerals - unknown spots
Elodea
                                 1.767

                                 2.961

                                 4.728
Fish
                                0.125

                                0.106

                                0.231
Mosquito    Snail
                                0.0452

                                0.244

                                0.289
                                                                                               0.658
                               0.185

                               0.843

                               0.544

                               1.387

-------
                    Table 17

Concentrations (ppm) of alachlor and metabolites
          in water in a model ecosystem
Compound
£>
alachlor
II
III
IV
V
VI
VII
VIII
IX
Extractable 14C
Unextractable ^C
Grand Total ^C
2.1 Silica Gel GF-254,
^_* *D x"**ti «i T-I TT*I ivmA v O 1 f* — tii
a/
R — Unhydrolyzed Water Hydrolyzed Water
0.70
0.61
0.51
0.43
0.33
0.27
0.20
0.13
0.07
0.00


0.000271
0.00105
0.00190
0.0105
0.000777
--
0.000832
0.00172
0.000687
0.000362
0.0181
0.0414
0.0595
me thanol -benzene, 5:95 by volume
i^I>-T"» fwsivi c* v*r\ 4~ r*
0.000677
--
0.00154
0.00364
0.00146
--
0.00854
--
0.00300
0.00465
0.0212
0.0118
0.0330
                      32

-------
                                              Table 18

                    Concentrations (ppm) of propachlor and metabolites in organisms
                                         in a model ecosystem
Compound.
D /
ii
propachlor
III
IV
V
VI
VII
VIII
Extractable 14C
Unextractable ^4(
Grand Total 14C
R / A i O 3 O f T OTn T^ £1 fl^lT"! ^ 3
i?— AXgcLc Vji.a.Ul i/a.pLirHa.
0.69
0.62
0.55
0.40
0.30
0.27-0.15
0.10
0.03
0.00
0.0211 0.00619 0.00930
] 0.186 0.00886 0.0476
0.207 0.0150 0.0569
Elodea Fish Mosquito Snail
--
_-
--
--
0.0154
--
--
--
0.0595
0.00243 0.00605 0.0264 0.0749
0.0869 0.00854 0.134 0.177
0.0893 0.0146 0.160 0.252
£/ Silica Gel GF-254, methanol-benzene, 5:95 by volume
—' Roman numerals - unknown spots

-------
                     Table 19

Concentrations (ppm) of propachlor and metabolites
           in water in a model ecosystem
Compound
I^7
II
propachlor
III
IV
V 0
VI
VII
VIII
Extractable ^C
Unextractable ^C
Grand Total 14C
^ Silica Gel GF-254
_' Roman numerals -
R — Unhydrolyzed Water Hydro ly zed Water
0.69
0.62
0.55
0.40
0.30
.27-0.15
0.10
0.03
0.00



--
0.0000614
0.0000564
0.00125
--
0.000144
0.0000830
0.0000216
0.0000382
0.00166
0.0121
0.0138
0.0000622
0.000108
--
0.00193
--
0.000277
0.000189
0.000297
0.00390
0.00676
0.00414
0.0109
, me thanol -benzene, 5:95 by volume
unknown s not
s

                       34

-------
                                  Bladdg)

  Bladex® is a member of the ^-triazine structural class of herbicides
  which has found use for the control of annual grasses and broadleaf
  weeds in corn as a preemergence treatment.  The preemergence treatment
  for corn with Bladej«y is preferred as injury often results when the
^herbicide is put on after the corn has emerged.  Under normal agricul-
  tural usage (1-4 Ib/A)  the half-life varied between 2-7 weeks, while
  laboratory half-life on three typical soils was about 15 days.  The
  toxicity to fish and birds is low, which correlates with the oral LD5Q
  of 334 mg/kg obtained for rats.

  The metabolism of Bladej® in rats (Hutson et: al., 1970) and degradation
  in soil have been examined (Beynon et al., 1972).  The principal
  metabolites in rat urine are the N-desethylated Bladex® and replacement
  of the 2-chloro group with an N-acetyl cysteine which is conjugated
  through a sulfur linkage.  The degradation in the soil is more compli-
  cated as the nitrile is hydrolyzed to the amide and, finally, the acid.
  The Cl group is replaced by OH and also N-desethylation takes place.

  The data for the distribution of Bladex® in the organisms and degrada-
  tion products are contained in Table 20.  With the exception of Elodea,
  which contained 0.621 ppm Bladex® and the crab with 0.172 ppm of the
  N-desethylated Bladex®, none of the other organisms had residues of
  identifiable metabolites.  The organisms contained extractable radio-
  activity which ranged from a low of 00.02 ppm for Daphnia to a value of
  0.629 ppm for Elodea.  Significantly, of this 0.629 ppm isolated from
  the water plant, nearly 99% was intact BladexB).  The level of
  unextractable radioactivity from the organisms averaged 48%, indicating
  a substantial degree of degradation of this herbicide.

  The water portion of this ecosystem (Table 21) is interesting in terms
  of the distribution of  degradation products as several were present in
  indentifiable amounts along with BladexS/ which was  found at 3.2 ppb.
  Two others which were found were the amide at 1.42  ppb and the
  N-desethylated amide at 0.0568 ppb.   In addition to these degradation
  products, there were several unidentified materials at levels of
  0.014-0.028 ppt.  Clearly,  none of the metabolites  isolated from the
  water appears in any of the organisms, which indicates that neither
  BladexB/nor any of its  metabolites  accumulate in the organisms  of  this
  model ecosystem.
                                 Bentazor
  Bentazor® is  a new herbicide that  is  being developed as  a broadleaf
  weed control  for application as  a  postemergence treatment.   There is
  little published information about the metabolism of this herbicide or
  its behavior  in the soil environment.   Recently,  a study was conducted
  to determine  the affinity of BentazoiUy for 12  Illinois soils and the
  data indicate that it  does not bind tightly to any of them (Abernathy
  and Wax,  1973).
                                   35

-------
                                               Table 20
     Compound
Bladex®
A

B

C



II

III

Extractable ^C

Unextractable ""C

Grand Total 14C
                     Concentrations  (ppm) of BladexS) and metabolites  in organisms
                                         in a model  ecosystem
St 1
n ci /
0.55
0.47
0.37
0.26
0.16
0.07
0.00



Algae Crab
__
0.172
__
__
_.
0.0579
0.0812
0.129 0.311
0.127 0.209
0.256 0.520
Daphnia Elodea
0.621
__
__
__
__
__
0.00818
0.0196 0.629
0.0202 0.0253
0.0398 0.654
 Fish
0.0354

0.0157

0.0511
Mosquito    Snail
 0.0277

 0.0751

 0.103
0.0454

0.0624

0.108
2.  Silica Gel GF-254, methanol-acetone-chloroform, 5:45:50 by volume
—' Roman numerals - unknown spots
A  2-chloro-4-amino-6-(l-methyl-l-cyanoethylamino) -j^-triazine
B  2-chloro-4-ethylamino-6-(l-methyl-l-carboxamidoethylamino)-£-triazine
C  2-chloro-4-amino-6-(l-methyl-l-carboxamidoethylamino) -s.-triazine

-------
                               Table 21
            Concentrations  (ppm) of Blades® and metabolites
                     in water in a model ecosystem
     Compound
Unhydrolyzed Water
Hydrolyzed Water
0.55
0.47
0.37
0.26
0.16
0.07
0.00



0.00321
0.0107
0.000142
0.0000568
0.0000142
0.0000142
0.0000284
0.0142
0.00359
0.0178
—
--
--
--
0.0000626
0.0000726
0.0000250
0.00016
0.00357
0.00373
A

B

C

I

II

III

Extractable

Unextractable

Grand Total 14C


a/
—' Silica Gel GF-254, methanol-acetone-chloroform, 5:45:50 by volume
—  Roman numerals - unknown spots
A  2-chlor-4-amino-6-(l-methyl-l-cyanoethylamino) ^s_-
B  2-chloro-4-ethylamino-6-(l-methyl-l-carboxamidoethylamino)-s_-triazine
C  2-chloro-4-amino-6-(l-methyl-l-carboxamidoethylamino)-j3-triazine
                                 37

-------
The data for the distribution of metabolites and  degradation products
of Bentazoi® in the organisms of the model ecosystem  are  compiled  in
Table 22.  The crab was the only organism that  contained  metabolites,
including BentazorUV at 0.512 ppm, anthranilic acid  at 1.27  ppm and
14-isopropyl anthranilamide at 0.622 ppm.  Most  of the other organisms
contained small amounts of W-C  labeled  extractable  metabolites that
were in such low concentration  identification was not possible.

The data for the degradation products  in the water  segment  of the  eco-
system are collected  in Table 23.  The  water contained 0.0213 ppm  of
IJ-isopropyl anthranilamide and  0.0505  ppm of Bentazon®, but no
anthranilic acid.   Several metabolites  and Bentazon®were accumulated
from the water to a slight extent by the crab as  both N-isopropyl
anthranilamide and Bentazori® were higher in concentration  in the crab
than in the water.

                                Dicamba

Dicamba is an effective'herbicide for  numerous  annual grasses and
broadleaf weeds in  grain  crops.  It  is  more persistent in the soil than
2,4-D which was essentially degraded in sandy  loam  and loam after  two
weeks; but dicamba  after  12 weeks was  still highly  active (Friesen,
1965).  Dicamba in  high organic soils  is detoxified rapidly at pH  5.3,
but appears to be highly  persistent  at  pH 7.5.  An  explanation for this
is the increase proliferation of soil  bacteria  at the lower pH which
are capable of degrading  the herbicide  (Swanson,  1969).   This herbicide
is not toxic to the rat,  LD5Q 1,028 mg/kg, nor  to rainbow trout as the
24 hr TLm  is 35,000 ppm  (Herbicide Handbook,  1970).

The metabolism of dicamba has been  investigated in  wheat  and bluegrass,
and the major degradation product is 5-hydroxy  dicamba which then  is
conjugated  (Broadhurst e_t a±,,  1966).   Further; transformations in
these two  plant species  include conjugation of  dicamba and
()-demethylation to  give  3,6-dichlorosalicylic  acid.  Metabolism studies
of dicamba with ring-labeled  l^C dicamba in rats  demonstrate that  this
herbicide  is rapidly  excreted in the urine with approximately 20%  of
the urinary metabolites  in  the  form of glucuronic acid conjugates (Tye
and Engel,  196_7) .   Feeding  studies  with dicamba in  rats reveal that
most residues were  in the aqueous portion of  the  body burden and not in
fatty tissues,  indicating this  herbicide is not stored (Tye and Engel,
1967).

The  picture  that  emerges  from examination of dicamba  in the model  eco-
system  for  the  organisms  as  seen in Table 24  is that  none of the
organisms  contains  residues  of  identifiable metabolites.  Only the crab
contains  0.743  ppm  of an unidentified  conjugate,  while the  rest of the
organisms  have  small  amounts  of extractable radioactivity and somewhat
larger  amounts  of unextractable radioactivity.   Clearly,  dicamba or  its
metabolites  do  not  accumulate  in the organisms  of this model ecosystem.
                                  38

-------
                                                     Table  22
vo
                            Concentrations  (ppm)  of  Bentazofl^and metabolites  in organisms
                                                in a model  ecosystem
Compound
A
B
BentazonB)
iV
Extractable 14>C
Unextractable ^C
Grand Total 14C
Rf— ' Algae
0.77
0.63
0.52
0.00
0.109
0.759
0.868
Clam
--
--
--
--
0.032
0.021
0.054
Crab
0.622
1.266
0.510
0.327
2.72
3.08
5.80
Daphnia
--
--
--
--
0.182
0.407
0.98
Elodea
--
--
--
--
0.092
0.168
0.26
Fish
--
--
--
--
0.012
0.036
0.048
Mosquito
--
--
--
--
0.146
0.716
0.86
Snail
--
--
--
--
0.084
0.378
0.462
      —/Silica  Gel  GF-254,  benzene-ethanol,  60:40  by  volume
      —'Roman numerals  -  unknown  spots
      A  N-isopropyl anthranilamide
      B  anthranilic acid

-------
                               Table 23

           Concentrations (ppm) of BentazonB/and metabolites
                     in water in a model ecosystem
a/
Compound Rf— Unhydrolyzed Water
A 0.77
B 0.63
Bentazoi® 0.52
]£/ 0.00
Extractable ^C
Unextr actable l^C
Grand Total 14C
0.000691
--
0.000281
0.000028
0.001
0.114
0.116
Hydrolyzed Water
0.0207
--
0.0502
0.00303
0.074
0.44
0.118
—I Silica Gel GF-254, benzene-ethanol, 60:40 by volume
.k/ Roman numerals - unknown spots
A  N-isopropyl anthranilamide
B  anthranilic acid
                                 40

-------
The water portion of this model ecosystem  (Table 25) contains  small
amounts of unconjugated dicamba, 0.000289 ppm, and about  l,800x  that
amount in the hydrolyzed water, 0.162 ppm.  The dicamba isolated after
hydrolysis was a conjugate which was released through the acid treat-
ment.  Also there were small amounts of 5-hydroxy dicamba, 0.0185 ppm,
released on treatment of the water with acid.  The tendency for  this
herbicide to become conjugated in the water portion of this ecosystem
is exemplified by the comparative figures for unextractable/extractable
i^C.  For the unhydrolyzed water, >99% was in the form of unextractable
radioactivity, while in the hydrolyzed water about 9970 became  extract-
able after acid hydrolysis.  In the hydrolyzed extract 89% of  the
radioactivity was dicamba.

                    2,4-dichlorophenoxyacetic acid

The herbicidal properties of 2,4-dichlorophenoxyacetic acid (2,4-D and
its esters) have been known for over 30 years.  In view of the lengthy
use of this herbicide and the availability of an excellent review of
this class of herbicidal chemicals (Loos, 1969), few comments will be
made about their mode of action.  In general, metabolism  and degradation
studies that have been carried out in plants indicate three generalized
metabolism pathways, namely, ring hydroxylation, side chain degradation
and conjugation of the acid or phenolic moieties (Loos, 1969).  Princi-
pal hydroxylated metabolites found in plants of 2,4-D are 2,3-dichloro-
4«»hydroxyphenoxyacetic acid and 2,5-dichloro-4-hydroxyphenoxyacetic
acid which result from a chlorine shift during hydroxylation.   The side
chain metabolism has been demonstrated through the use of ^C  labeled
2,4-D and trapping of the ^C02 produced by oxidation of  the side chain.
Finally, conjugates of the ring hydroxylated metabolites  and intact
2,4-D with glucose appear to be an important mode of detoxication of
this herbicide by plants (Loos, 1969).  In addition to the characterized
metabolites, numerous unidentified metabolites from plants have been
observed and corroborate the observation from the present model
ecosystem work that this chemical can undergo substantial degradation.

The degradation of 2,4-D in the soil by microorganisms is optimal in
warm, moist soil.  Autoclaving of the soil destroys the bacteria and
reduces the rate of degradation of 2,4-D.  Primary degradation routes
of the phenoxyacetic acids appear to go in two stages,  namely,
degradation to the phenol and then degradation of the phenol to carbon
dioxide and water (Loos, 1969).

With regards to the environmental accumulation of 2,4-D by fish in an
aquatic food web, a recent study (Schultz,  1973) has provided  insight
into the uptake of this herbicide by fish.   In general, the three
species of fish exposed to this herbicide did not accumulate it over
the concentration in the water.  As would be expected,  fish exposed to
the herbicide in water held at pH 9 contained less  residues of 2,4-D
than those exposed at pH 6.  Further, less  than 10% of the radioactivity
accumulated by the fish was parent material and the major metabolite
                                  41

-------
                                                     Table 24
S3
                           Concentrations (ppm) of dicamba and metabolites in organisms
                                               in a model ecosystem
Compound
dicamba
A
B
Extractable 14C
Unextractable ^C
Grand Total 14C
Rf-' Algae
0.86
0.38
0.04
0.228
1.390
1.618
                                              Clam   Crab
                 Daphnia  Elodea
Fish
Mosquito  Snail
        0.743

0.0128  0.743

0.0144  0.374

0.0272  1.117
      —' Whatman No. 1 filter paper, benzene-acetic acid, 2:1 by volume
      A  3,6-dichloro-5-hydroxy-2-methoxy benzoic acid
      B  Conjugated metabolite
0.000
0.167
0.167
0.325
0.593
0.918
0.00665
0.0122
0.0188
•* •»
0.0736
0.281
0.355
0.0720
0.252
0.324

-------
                               Table 25

            Concentrations (ppm) of dicamba and metabolites
                     in water in a model ecosystem
     Compound

d icamba

A

B

Extractable

Unextractable
Grand Total
            14,
RjS'
0.86
0.38
0.04



Unhydrolyzed Water
0.000289
--
0.0000114
0.000300
0.163
0.163
Hydro lyzed Water
0.162
0.0185
0.000181
0.181
0.0022
0.183
a/
—' Whatman filter paper No. 1, benzene-acetic acid, 2:1 by volume
A  3,6-dichloro-5-hydroxy-2-methoxy benzoic acid
B  Conjugated metabolite
                                43

-------
was the glucuronic acid conjugate of 2,4-D.  The small amounts of 2,4-D
found in the fish demonstrate that the fish can metabolize this
herbicide.

The model ecosystem experiment confirmed the earlier work regarding  the
propensity for 2,4-D to undergo substantial degradation and not accumu-
late in aquatic organisms.  For example in Table 26 for the organisms,
there were no residues of 2,4-D and seven unidentified metabolites.
The substantial numbers of unidentified metabolites are disturbing as
six metabolites, including 2,4-D, were co-chromatographed with the
extracts  from these organisms.  The compounds  chromatographed were
2,4-D, 2,3-dichloro-4-hydroxyphenoxyacetic acid, 2,4-dichloro-5-
hydroxyphenoxyacetic acid, 2,5-dichloro-4-hydroxyphenoxyacetic acid,
2,4-dichlorophenol, 3,5-dichlorocatechol, 2,3-dichlorohydroquinone and
2,3-dichlororesorcinol.

The water portion of the model ecosystem experiment (Table 27) had
numerous  spots which did not  correspond to any of  the chromatographed
metabolites.  Interestingly,  the water primarily contained metabolites
which had Rf values of 0.58  or  less, while the snail, algae and Elodea
contained metabolites  and/or  degradation products  with higher Rf values.
This is expected as polar  products remain  in  the water and  less polar
materials partition into the  organisms.  Clearly,  2,4-D does not
accumulate  in any of the organisms of  this model ecosystem  and appears
to be degraded  into water  soluble compounds.

                                Pyrazon

Pyrazon  is  a herbicide used  for the  control of annual broadleaf weeds
in sugar  beets  and red beets. The usual modes of  application are  as a
preemergence, broadcast or early  postemergence as  a banded  treatment at
2-4  Ib/A.  This herbicide  is  relatively nontoxic   to mammals as the  oral
LD5Q to  the rat  is 3,000 mg/kg  (Anon.  1973).   It does not leach readily
through  clay, clay  loam or sandy  clay  and  loam soils and has a soil
persistence time of about  1-2 months  (Frank,  1972).  The mode of action
in the susceptible plant,  lambsquarters, is  inhibition of the Hill
Reaction which  is  important  in  photosynthesis. However,  in vivo com-
parison  of  the  metabolic rates  of sugar beets, which are nonsusceptible,
and  lambsquarters, which are susceptible,  shows  that pyrazon accumulates
 in the stems  and  leaves  in the  susceptible species, while in the
tolerant sugar  beets  the pyrazon  is  rapidly degraded to nonphytotoxic
species.  Hence,  the  selectivity  in  vivo  in  the  comparison  of sugar
beets  and lambsquarters  lies in the  greater ability of the  sugar beets
 to degrade  pyrazon  (Frank, 1972).

The  metabolism  of  pyrazon  in plants  and  soil  has been examined.  In
 sugar  beets the herbicide  was degraded to  three  primary metabolites,
 two of which were  positively identified.   The two  identified were  the
 conjugate with  the  free  amino group  with glucose  to  form N-glucosyl
 pyrazon  and desphenyl  pyrazon.   In  the soil  the  only metabolite
 identified  was  the  desphenylated  pyrazon  (Stephensen and Ries,  1969).
                                  44

-------
                  Table 26
Concentrations (ppm)  of 2,4-D and metabolites
      in organisms in a model ecosystem
Compound
.*/
II
III
VI
V
X
XII
Extractable ^C
Unextractable 14-C
Grand Total 14C
^ Silica Gel GF-254,
_' Roman numerals - 111
RfS'
0.97
0.89
0.80
0.65
0.58
0.10
0.00



Algae
0.282
1.0295
0.477
0.377
0.295
1.675
1.362
5.498
17.625
23.123
benzene-dioxane-acetic acid,
ilrnown snof-R
Elodea
0.178
0.456
0.447
--
--
0.768
0.873
2.722
7.755
10.477
90:25:4 by volume
                                                Fish
                                                0.0431



                                                0.00226

                                                0.0454

                                                0.211

                                                0.256
Snail

0.285



0.301
0.171

0.757

6.421

7.178

-------
                               Table 27

            Concentrations (ppm) of 2,4-D and metabolites
                    in water in a model ecosystem
     Compound
VI

VII

VIII

IX

X

XI

XII

Extractable
14r
Unextractable

Grand Total
                    Unhydrolyzed Water     Hydrolyzed Water
0.58
0.63
0.56
0.49
0.39
0.10
0.067
0.00



0.000029
0.0000914
0.0000658
--
--'
--
--
0.00000496
0.000191
0.0128
0.0130
0.0000351
0.00260
0.00205
0.00226
0.000417
0.000474
0.000271
0.000180
0.000829
0.0120
0.0128
—  Silica Gel GF-254, benzene-dioxane-acetic acid, 90:25:4 by volume
—' Roman numerals - unknown spots
                                 46

-------
The data  for  the distribution  of metabolites  and degradation products
in the organisms of pyrazon  are in  Table  28.  Again,  as in the case of
the 2-chloroacetanilides, no identifiable compounds with the exception
of the crab,  0.476 ppm, had  residues  of either  pyrazon or degradation
products.  Further, very little of  the radioactivity  (extractable and
taaextractable) was isolated  from any  of the organisms (0.0573 ppm for
fish to 0.629 ppm for  crab), which  may be explained by the relatively
high water solubility  of pyrazon, 300 ppm (Anon.  1970).

The water portion of this ecosystem (Table 29)  contains five unidenti-
fied metabolites not found in  the organisms,  including pyrazon at
0.212 ppm and the desphenyl  pyrazon at 0.0714 ppb.  Clearly, pyrazon
shows a similar pattern to the 2-chloroacetanilides as  it is not
accumulated by the organisms of the model ecosystem,  but degrades to
polar, water  soluble metabolites.

                             Trifluralii®

Trifluralii®  , or treflan, is one of  many dinitroaniline herbicides
which include Cobesffi),  Nitralii® and Benefit®.   These  herbicides  have
found extensive use in soybeans to  control common annual grasses, pig-
weed and lambsquarters and are usually applied  as either a preplant  or
preemergence  application.  TrifluralirUv is best suited  for soils with
3% organic matter or less (Wax, 1973).  Extensive toxicological  data
obtained on Trif luralirQy indicate a low order of acute  toxicity, LD50
to rats of >10 g/kg, LC5Q of an emulsifiable  concentrate to bluegills,
fathead minnows and goldfish of 0.58  ppm,  0.94  ppm and  0.59 ppm,
respectively  (Parka and Worth, 1965).

The metabolism of TrifluralinD in plants,  animals and soil as  well as
photodecomposition has been  investigated.  In addition,  the anaerobic
degradation of this herbicide has been examined.  The pathways for
degradation of this herbicide have  been the subject of  an excellent
review (Probst and Tepe, 1969) and, therefore,  will only be discussed
briefly.  Degradation  of Trif luraliiilP in  the  soil under  aerobic  con-
ditions leads to numerous products, including the mono-  and di-
dealkylated derivatives of Trifluralin®,  as well as mono-  and  di-
dealkylated TrifluralinIP with one of  the  2,6-nitro groups  reduced to an
amine.  A final metabolite isolated from  the  aerobic  system is the
conjugated a,Q!,Q!-trifluorotoluene-3,4,5-triamine.

The experiment with Trifluraliifiywas carried out twice employing
different modes of application.  The  normal-use pattern  of Trifluralin®
is a soil treatment, so the  analogous treatment in the model ecosystem
would be treatment of  the sand at evenly  spaced intervals  (2.5 cm
apart) with 5 mg which corresponds  to about 1 Ib/A.   Since sorghum is
susceptible to Trif luraliiiB', a common variety of soybeans  was  grown  in
place to establish the viability of the sand  for plant growth.   While
the Trifluraline) in this case was a sand  treatment, no  caterpillars
were added to act as the first stage of the food chain as  well as  a
dispersing agent.  The sand  treatment was  then  compared  to the normal
                                 47

-------
•p-
00
                                                      Table  28



                            Concentrations  (ppm)  of  pyrazon  and metabolites  in organisms

                                                in a model ecosystem
Compound
pyrazon
A
I^7
II
III
IV
V
Extractable ^C
Unextractable ^
Grand Total 14C
R -' Algae
0.63-0.69
0.47-0.51
0.40-0.43
0.23
0.16
0.10
0.00
0.0758
C 0.131
0.207
Clam Crab Daphnia Elodea Fish
0.476
_-
__
__
__
_.
0.0233
0.0498 0.499 0.0536 0.105 0.0336
0.0180 0.130 0.0455 0.0552 0.0237
0.0678 0.629 0.0991 0.160 0.0573
Mosquito Snail
__
__
--
__
__
--
__
0.175 0.127
0.148 0.0592
0.323 0.186
      a/
      —  Silica Gel GF-254, benzene-ethanol, 60:40 by volume

      —' Roman numerals - unknown spot

      A  5-amino-4-chloro-3(2H)pyridazinone

-------
                               Table 29



            Concentrations (ppm) of pyrazon and metabolites

                     in water in a model ecosystem
Compound
                      R
,'      Unhydrolyzed Water
Hydrolyzed Water
pyrazon
A
it/
II
III
IV
V
Extractable ^C
Unextractable 14C
Grand Total 14C
0.63-0.69 0.0112
0.47-0.51
0.40-0.43
0.23
0.16
0.10
0.00 0.0000300
0.0112
0.0203
0.0315
0.00998
0.0000714
0.0000430
0.0000260
0.0000471
0.0000764
0.000106
0.0103
0.0105
0.0208
a/
—' Silica Gel GF-254, benzene-ethanol, 60:40 by volume

—' Roman numerals - unknown spots

A  5-amino-4-chloro-3(2H)pyridazinone
                                49

-------
sorghum* treatment which is the standard mode of application of
pesticides in the model ecosystem.  The discussion of Trifluralii® will
then engender a comparison of the two methods of application and  its
effect on the persistence and degradation pattern of this herbicide.

In Tables 30 and 32 are the data for the distribution of the metabo-
lites in the organisms of the sand- and sorghum-treated Trifluralii^
ecosystems.  Several interesting differences are related to the two
treatments as the snail and fish contain more Trifluralin§) in the  sand-
treated system than in the sorghum-treated system.  This is shown  by
the data which indicates that the snail and fish contained about 5.4x
and 3.Ox more herbicide in the sand experiment.  Another striking
difference is the lack of algae in the sorghum treatment and the
existence of algae in the sand-treated ecosystem.  The caterpillar,
which is the dispersing agent and the first member of the food chain,
apparently spread residues of Trifluralinfi) more efficiently into the
water which were phytotoxic and, therefore, inhibited the growth of
the algae in the sorghum treatment.

The data for the water portion of the model ecosystem (Tables 31 and
32) support the role of the caterpillar in spreading the TrifluralinS*
and metabolites in the system as the total radioactivity in the water
at the end of the experiment was about 6.4x greater in the sorghum-
treated ecosystem as compared with the sand-treated system.  Apparently,
the TrifluralinB) is adsorbed on the sand and therefore does not
dissolve in the water.  In contrast to the approximately 6-fold
difference in total radioactivitvin the water of the sorghum-treated
system, the amount of TrifluralinS' in the water is only 1.5x greater
in the sorghum treatment than in the sand treatment.  Finally, the
concentration of TrifluralirtB) by the organisms appears to be affected
by the type of treatment as the values for accumulation of TrifluralinS'
over  the concentration in the water by the fish and snail in the  sand
treatment is 4,200x and 153,000x, respectively.  In the sorghum treat-
ment  the values for the fish and snail are 930x and 17,700x,
respectively.  The caterpillar which degrades TrifluraliiQv in the
sorghum treatment, as well as the opportunity for TrifluralinS' to
undergo photodecomposition on the sorghum leaves before consumption by
the  caterpillar, have a substantial effect on the persistence of  this
herbicide as demonstrated by  the accumulation factors for the snail
and  fish.  The accumulation factors of TrifluralinR for the fish  and
snail are  in the same order of magnitude as that obtained for
methoxychlor which was accumulated »vl,500x by the fish and about
123,000x by the snail  (Metcalf et_ al., 1973).  The snail is incapable
of degrading even the most biodegradable DDT analogue, methoxychlor,
as well as the herbicide, Trifluralin®.
                                  50

-------
                                                      Table  30
Ul
                          Concentrations  (ppm)  of Trifluralii® and metabolites  in organisms
                                    in a  model  ecosystem after sorghum treatment
Compound
.Trifluralii®
1^
A
C
II
III
IV
E
V
VI
VII
VIII
Extractable l^C
Unextractable l^C
Grand Total ^C
Rf£' Daphnia
0.74
0.51
0.39
0.32
0.24
0.20
0.17
0.13
0.11
0.07
0.04
0.00
0.445
1.017
1.462
Fish Mosquito Snail
0.261 — 5.046
--
0.337
--
0.0399
0.216
--
--
--
0.228
__
0.506 — 0.796
0.767 0.238 6.535
1.011 0.520 6.648
1.777 0.758 13.183

-------
                                            Table 30 (con't.)

I/
   Silica Gel GF-254, hexane-acetone-MeOH,  90:10:2  by volume
—  Roman numerals - unknown  spots
A  a,Q!jQ;-trif luoro-2 ,6-dinitro-N-propyl-£-toluidine
C  2,6-dinitro-4-trif luoroariilir.e
E  2-ethyl-5-trifluoromethyl-7-nitrobenzimidazole

-------
                               Table  31

         Concentrations  (ppm)  of Trif luraliifD and metabolites
        in water in a model  ecosystem after sorghum treatment
                                Unhydrolyzed Water
     Compound



Trifluralin®

I

A

C

II

III

IV

E

V

VI

VII

VIII

Extractable 14C

Unextractable ™

Grand Total
—' Silica Gel GF-254, hexane-acetone-MeOH, 90:10:2 by volume
—' Roman numerals - unknown spots
A  a,0!,0!-trif luoro-2,6-dinitro-N-propyl-jD-toluidine
C  2,6-dinitro-4-trifluoroaniline
E  2-ethyl-5-tr if luommethyl-7-nitrobenzimidazole
Hydrolyzed Water
0.74
0.51
0.39
0.32
0.24
0.20
0.17
0.13
0.11
0.07
0.04
0.00



0.000282
0.000066
0.000087
0.000374
0.000322
0.000139
0.000315
0.000475
0.000415
0.000928
0.00123
0.00526
0.00989
0.0290
0.0388
—
--
--
--
--
--
0.000199
0.000328
0.000271
0.000486
0.000800
0.0116
0.0137
0.0253
0.0290
                                 53

-------
                                                     Table 32
Ui
                         Concentrations (ppm) of Trif luralin® and metabolites in organisms
                                    in a model ecosystem after sand treatment
Compound
Trifluralin®
&
A
B
D
E
V
Extractable l^C
Unextractable 14C
Grand Total 14C
RfS' Algae
0.74
0.61
0.56
0.36
0.22
0.09
0.00
0.350
0.475
0.825
Daphnia Fish
0.775
0.167
0.179
--
0.04
0.052
0.138
0.317 1.323
0.311 0.129
0.628 1.452
Mosquito Snail
27.085
0.667
__
0.323
__
__
0.766
0.208 28.870
0.168 0.183
0.376 29.053
      £  Silica Gel GF-254,  hexane-acetone-MeOH,  90:10:2 by volume
      £/ Roman numerals - unknown spots
      A  a,a,Q;-trifluoro-2,6-dinitro-N-propyl-£-toluidine
      B  N,N-dipropyl-3-nitro-5-trifluoromethyl-£-phenylenediamine
      D  2-ethyl-5-trifluoromethyl-7-nitro-l-propylbenimidazole
      E  2-ethyl-5-trifluoromethyl-7-nitrobenzimidazole

-------
                                Table  33

         Concentrations  (ppm)  of TrifluralinS) and metabolites
          in water in a model  ecosystem after sand treatment
Compound
Tr£fluralin5)
iV
A
II
B
C
D
III
E
IV
V
Extractable ^C
Unextractable ^C
Grand Total ^C
R a/
Rf~
0.74
0.61
0.56
0.48
0.36
0.27
0.22
0.14
0.09
0.05
0.00



Unhydrolyzed Water
0.000184
0.0000343
—
--
--
0.000140
0.0000917
0.000111
0.000414
0.000597
0.00144
0.00301
0.00383
0.00684
Hydro ly zed Water
--
--
0.0000104
--
--
--
--
0.000017
0.0000737
0.000299
0.000687
0.00109
0.00274
0.00383
2.  Silica Gel GF-254, hexane-acetone-MeOH, 90:10:2 by volume
—' Roman numerals - unknown spots
A  0!,a,Q!-trif luoro-2,6-dinitro-IJ-propyl-j>-toluidine
B  N,N-dipropyl-3-nitro-5-trifluoromethyl-£-phenylenediamine
D  2-ethyl-5-trifluoromethyl-7-nitro-l-propylbenimidazole
E  2-ethyl-5-trifluoromethyl-7-nitrobenzimidazole
                                  55

-------
                              SECTION V

           EXAMINATION OF A MITICIDE, SELECT PLASTICIZERS,
                   A FUNGICIDE AND A BACTERIOSTAT

The final segment of this report of the  fate of select pesticides  in
the aquatic environment deals with a miticide, two types of  plasti-
cizerSj a fungicide and a bacteriostat.  The miticide was chosen as
little environmental information is known about acaricides in general
and further, this is a new class of miticide which has not yet  found
generalized use.  Therefore, it would be of interest to examine the
persistence and uptake of this pesticide by the organisms of the model
ecosystem before it is used  in large quantities in the environment.

Two types of plasticizers were examined  in the model ecosystem  because
they have been demonstrated  to be pollutants of fish, birds  and many
other  forms of wildlife.  In contrast to the previously discussed
pesticides, these compounds  were not purposefully applied to control a
pest,  but have become pollutants unintentionally as the result  of
either industrial discharge  during production or from seepage into the
environment during their intended use.   The high lipid solubility,
inertness to chemical and biological degradation, particularly  for the
higher chlorinated PCB's, and coupled with their multibillion pound
production clearly explains  the worldwide occurrence of these materials
in the environment.

The examination of captan in the model ecosystem was important  for two
reasons.  First, the use of  fungicides containing mercury is now
generally considered to be both environmentally unsafe as well  as
hazardous to human health.   The  mercury fungicides are quite toxic to
humans and can cause severe  poisoning when ingested.  Some adults  and
children in New Mexico and Iran mistakenly utilized grain treated  with
mercury fungicides for feed  for swine and for making bread and, as a
result, they suffered from acute mercury posioning.  Secondly,  captan
has been used  for about 20 years without apparent environmental prob-
lems related to persistence  and food web magnification.  In  order  to
validate the use of the model ecosystem  as a useful tool for prediction
of pesticide persistence in  food  chain accumulation, it is necessary to
examine compounds, such as captan, which  have had  little environmental
impact after widespread use, as well as  persistent  compounds, such as
dieldrin, DDT  or aldrin.  If both ends of the persistence spectrum do
not agree both in  the field  and the model ecosystem, then this  system
would  be of  little value as  a predictive tool for new pesticides.

                              Banomite®

Banomite® is a new class of  miticidal benzoyl chloride phenylhydrazines
which  is being developed  for the  control of mites which attack  citrus
 (Kaugers ejt  al. ,  1973: Moon  et  al.,  1972).  Since these are  a new  class
of acaricides,  environmental information or metabolism data  on  these
 compounds has  not been published.
                                  56

-------
The data contained in Table 34 describe the distribution  of  BanomiteS'
and its degradation products in the organisms of the model ecosystem.
Three of the organisms contain small residues of Banomitely,  in  particu-
lar the crab, 0.0156 ppm; Elodea, 0.041 ppm and the mosquito larvae
with 0.0736 ppm.  The only other metabolism or degradation product
ffiat could be identified in the organisms is the hydrolysis  product  of
BanomiteJy, benzoic acid 2,4,6-trichlorohydrazide.  The algae, crab,
Elodea, mosquito and snail contained 0.142 ppm, 0.0364 ppm,  0.0515 ppm,
0.300 ppm and 0.306 ppm, respectively, of this metabolite.   In  addition
to the two identified compounds, there were eight other unidentified
metabolites.  This acaricide is labile and undergoes degradation  to  the
numerous metabolites isolated from the organisms.  The snail and  fish
contain considerable amounts of unidentified Metabolite II with 10.69
ppm and 1.62 ppm, respectively.

Examination of the data for the water (Table 35) for BanomiteB)  shows a
similar distribution of Banomite*' and degradation products.  Therewere
small amounts of the two characterized compounds, namely, Banomite**'at •
18.62 ppt and benzoic acid 2,4,6-trichlorophenylhydrazide at 294  ppt.
The value for the water concentration of BanomiteB', when divided  into
those organisms which contained this miticide, reveal that the  crab,
Elodea and mosquito accumulated Banomite® from the water 840x,  2,200x
and 4,000x, respectively.  The other degradation product, or metabolite
which was found in the organisms, benzoic acid 2,4,6-
trichlorophenylhydrazide, was accumulated from the water by  the algae,
crab, Elodea, mosquito and snail 500x, 125x, 175x, l,000x and l,000x,
respectively.  However, the unidentified Metabolite II was accumulated
from the water by the fish 3,000x and the snail o/19,000x, and therefore
its structure should be investigated further to determine if this
metabolite has any physiological effect at this level.

                      Di-n-octyl phthalate (DOP)

The concern for environmental quality has stimulated interest in  the
disposition of substances which interfere with residue analysis,  such
as the polychlorinated biphenyls (PCB's) and the phthalate ester  plasti-
cizers.  The latter are produced in enormous quantities, for example,
some 4.5 x 10? Ibs in 1972 {Anon. 1972).  The phthalate esters,  which
as pollutants occur in fish taken from streams and rivers in the United
States (Mayer et al., 1972), appear to have a low order of acute
toxicity to aquatic organisms (Sanders et al., 1973);  but, for instance,
di-2-ethylhexyl phthalate,  the largest produced phthalate plasticizer,
accumulates in snails, mosquito larvae and mosquito fish in a model
ecosystem experiment (Metcalf et _al., 1973).  Therefore, to establish
the relationship between plasticizer chemical structure and environ-
mental persistence, di-n-octyl phthalate was examined in the model
ecosystem.

Previous model ecosystem studies with di-2-ethylhexyl phthalate  (DEHP)
demonstrated that this phthalate ester was degraded to mono
2-ethylhexyl phthalate and phthalic acid (Metcalf et_ al., 1973).  Only
                                 57

-------
00
                                                     Table 34

                           Concentrations (ppm) of BanomiteQy and metabolites  in
                                                in a model ecosystem
organisms
Compound
Banomite®
&
II
III
A
IV
V
VI
VII
VIII
Extractable 14C
Unextractable 14(
Grand Total 14C
R
0
0
0
0
0
0
0
0
0
0
-
1
J

.£='
.75
.61
.53
.35
.27
.21
.14
.07
.03
.00



Algae
--
--
0.963
0.181
0.142
0.202
0.269
--
0.514
0.923
3.191
6.602
9.793
Clam
0
--
2.044 0
0
0
—
0.227 0
--
0
0.0703 1
2 . 342 1
0.136 1
2.478 2
Crab Daphnia
.0156
__
.0670 0.686
.0568
.0364
__
.0406
—
.140
.00850 0.255
.366 0.941
.0878 0.877
.454 1.818
Elodea
0
0
1
0
0

0

0
0
1
1
3
.0410
.0928
.048
.0662
.0515
--
.185
--
.059
.328
.874
.445
.319


1
0


0
0

0
2
0
2
Fish
--
--
.624
.378
--
--
.0943
.0452
--
.180
.323
.451
.774
Mosquito
0.0736
0.266
0.453
0.265
0.300
--
0.324
0.286
0.533
1.652
4.156
2.137
6.293
Snail
--
0:565
10.685
1.160
0.306
--
2.549
0.938
0.478
0.872
17.542
1.777
19.319
       £  Silica Gel GF-254, n-hexane-ethyl acetate, 80:20 by volume
       —'  Roman numerals - unknown spots
       A  benzoic acid   2,4,6-trichlorophenyl hydrazide

-------
                               Table 35

           Concentrations (ppm) of Banomite® and metabolites
                     in water in a model ecosystem
     Compound
                        a/
Unhydrolyzed Water
Hydrolyzed Water
Banomite®
£/
II
III
A
IV
V
VI
VII
VIII
Extractable ^C
Unextractable l^C
Grand Total 14C
0.75
0.61
0.53
0.35
0.27
0.21
0.14
0.07
0.03
0.00



0.00000892
0.0000602
0.000444
0.000452
0.000141
0.000110
0.000693
0.00180
0.000662
0.00176
0.00612
0.0243
0.0304
0.0000097
0.0000328
0.0000948
0.000688
0.000153
0.000184
0.00157
0.000682
0.00362
0.0093
0.0164
0.00599
0.0224
—'  Silica Gel GF-254, n-hexane-ethyl acetate, 80:20 by volume
—'  Roman numerals - unknown spots
A   benzoic acid-2,4,6-trichlorophenyl hydrazide
                                59

-------
the mosquito larvae, snail and mosquito fish accumulated  from  the water
DEHP 100,000x, 21,000x and 130x, respectively.

The data for the distribution of DOP in the organisms of  the model
ecosystem are collected in Table 36.  The algae  contained the  most DOP
at 1.80 ppm, the snail with the next highest at  0.85 ppm,  the  fish and
mosquito at 0.59 ppm and Daphnia at 0.16 ppm.  In addition to  DOP there
were about six unidentified metabolites, but no  mono octyl phthalate
was isolated from any of the organisms.  In all  cases, except  the fish,
most of the radioactivity averaged about 75% unextractable, whereas the
fish had only about 22% unextractable.  Perhaps  the low titer  of liver
microsomal enzymes  in Gambusia affinis  (Krieger  and Lee,  1973)  accounts
for the inability of the fish to convert DOP to  polar unextractable
products.

The water portion of this model ecosystem experiment as indicated in
Table 37 contained  small amounts of DOP, 6.3 ppt, no mono octyl
phthalate and small amounts of phthalic acid,  0.782-1.82  ppb.   Using
the values obtained for the amounts of DOP in  the water and organisms,
concentration factors can be calculated for algae 28,500x, Daphnia
2,600x, fish and mosquito larvae 9,400x and 13,600x for the snail.
Previous ecosystem  evaluation of the branched  DEHP indicated that this
plasticizer was accumulated from the water by  the snail 21,000x, the
mosquito fish 130x  and the mosquito larvae 100,000x.  With only two
esters examined in  this model ecosystem, it is difficult  to account for
the differences in  the uptake of these  structurally related plasticizers,
It  is obvious that  despite the susceptibility  of these two plasticizers
to undergo substantial degradation, their high lipid solubility
accounts for their  uptake and storage in the organisms of this model
ecosystem  (Mayer et al., 1972).

                       Polychlorinated  Biphenyls

The ubiquitous occurrence of polychlorinated biphenyls (PCB's)  in the
environment has been well reviewed  (Peakall, 1972) and, therefore, a
few brief  comments  will be made about their chemical and  environmental
properties.  The industrial PCB's are a complex  mixture of chlorinated
biphenyls which have a multitude of uses including heat-transfer fluids,
plasticizers and extenders for pesticides.  The  inert chemical proper-
ties of these compounds make them excellent in their numerous
applications, but,  because of  their environmental stability and high
lipid solubility, they have been found  in the  lipoidal tissues of
numerous animals, principally  in fish  (Henderson, 1971) and birds
 (Risebrough,  1969).  The primary production of polychlorinated
biphenyls  in  the United States is by the Monsanto Company under the
trade name Aroclor© 1254,  1242, 1221, etc., where the first two digits
identify it as a derivative of biphenyl which  has 12 carbons and the
last  two digits relate  to  the  average percent  chlorine, i.e.,  1254 has
54% chlorine.
                                  60

-------
                                               Table 36

                     Concentrations (ppm) of di-ri-octyl phthalate and metabolites
                                   in organisms of a model ecosystem
Compound
DOP
&
II
III
IV
VI
VII
Extractable 14C
Unextractable ^C
Grand Total 14C
R a/
0.92
0.73
0.53
0.47
0.35
0.25
0.00



Algae
1.7963
0.6709
0.3356
0.3797
0.0808
--
7.3654
10.6348
43.2011
53.8359
Daphnia
0.1645
--
0.1425
0.1722
--
--
1.2001
1.6793
7.4703
9.1488
Fish
0.5920
--
0.0357
0.0374
--
--
0.2649
0.9300
0.3974
1.3273
Mosquito
0.5925
0.1748
0.4855
0.1641
0.0906
--
0.3314
1.8389
6.7225
8.5614
Snail
0.8543
0.0545
0.1643
0.1477
--
0.0569
0.2882
1.5659
2.9077
4.4736
^•'l Silica Gel GF-254, benzene-acetone-petroleum ether-acetic acid, 50:5:25:1  by volume
—  Roman numerals, unknown spots

-------
                                Table 37




    Concentrations (ppm)  of di-ja-octyl phthalate and metabolites

                  in organisms of a model ecosystem
     Compound
Unhydrolyzed Water
Hydrolyzed Water
DOP
1^
II
III
IV
V
VI
phthalic acid
VII
Extract able ^C
Unextractable 1
-------
The initial studies with the PCB's and animals were  carried out with
the complex mixtures which make analysis of  the metabolites and change
in the constitution of the PCB mixture very  difficult.  However,  with
the recent availability of pure 14-C isomers  of three of the PCB's,  more
pr.e^ise studies on the uptake and metabolism of PCB's can be undertaken.
The three labeled PCB's currently available  from the Mallinckrodt
Chemical Company are 2 ,5,2'-trichlorobiphenyl, 2,5,2',5'-
tetrachlorobiphenyl and 2,4,5,2',5'-pentachlorobiphenyl.  Through  the
cooperation of Professor Robert L. Metcalf,  University of Illinois,  it
was possible that these compounds could be obtained  for model ecosystem
studies in his laboratory.

The data in Table 38 contain the distribution in both the water and
organisms for the three pure isomer l^C PCB's.  An interesting aspect of
the data is the gradual increase in concentration of the three com-
pounds as the chlorine number is increased from three to five.  For
example, the fish contains 1.28 ppm trichlorobiphenyl, 14.24 ppm
tetrachlorobiphenyl and 119.71 ppm pentachlorobiphenyl.  For the  snail
the increased accumulation of the biphenyls  is similar to the fish,  but
the levels are higher as 18.97 ppm trichlorobiphenyl, 47.33  ppm tetra-
chlorobiphenyl and 587.35 ppm for pentachlorobiphenyl were  isolated
from this organism.

Another significant aspect of the data is the gradual decrease in
percent unextractable radioactivity in the fish as the number of
chlorine substituents is increased from three to five.  The  percentage
values for the unextractable radioactivity are 15% for the  trichloro-
biphenyl, 2.44% for the tetrachlorobiphenyl  and 1.10% for the
pentachlorobiphenyl, which indicates the relative susceptibility  of
these three compounds to undergo metabolism  by the fish.

One last comment regards the relative accumulation from the water of
these PCB's by the snail and fish.   The fish accumulated the  3-, 4-  and
5-chlorinated biphenyls 6,400x, ll,863x and  12,153x, respectively, the
concentration in the water.   The snail showed a greater differential
between the numbers of chlorine substituents and accumulation values as
the 3-, 4- and 5-chlorinated biphenyls accumulated 5,795x,  39,439x and
59,629x, respectively, the water concentration.   This differential
between accumulation factors for the snail as compared to the  fish may
reflect the greater metabolic capacity of the fish to transform these
lipid soluble compounds to water soluble compounds after they are taken
up from their environment.

                                Captan

This nonmercury fungicide is widely employed as  an effective  agent for
the control of fungus in vegetables, fruits  and ornamentals.   In
addition, it is utilized as an effective seed treatment.  The  safety of
this fungicide to rats is demonstrated by its oral LD5Q of >15,000
mg/kg and an intravenous 11)50 of 50-100 mg/kg (Metcalf,  1971).  Feeding
studies conducted with captan for a 2-year period at 10,000 ppm with
                                 63

-------
                                               Table 38
                            Distribution of chlorinated biphenyls and their
                              degradation products in the model ecosystem
I.   2,5,2'-trichlorobiphenyl
     total 1*C

    Unknown I (Rf 0.66*)

    trichlorobiphenyl (Rf 0.56)

    Unknown II (Rf 0.23)

    Unknown III (Rf 0.10)

    Unknown IV (Rf 0.06)

    Unknown V (Rf 0.04)

    Unknown VI (Rf 0.03)

    Polar (Rf 0.0)

    Unextractable
                                                         chlorinated biphenyl equivalents - ppm
H20
0.03845
0.00015
0.00020
0.00005
--
0.00055
0.00040
0.00040
0.02265
0.01405
Oedoeonium
(alea)
23.2155
15.9575
1.4630
0.0520
~-
--
--
0.0685
0.5185
5.1560
Physa
(snail)
31.2015
18.9720
1.1590
0.6480
0.9735
0.5460
0.2205
0.4410
3.9315
4.3100
Culex
(mosquito)
2 . 7030
1.1995
0.1630
—
--
--
--
--
0.4795
0.8610
Gambus ia
(fish)
3.2055
0.2085
1.2800
0.1595
--
--
--
--
0.9985
0.5590

-------
                                                  Table 38 (con't.)
Oi
II. 2,5,2' ,5'-tetrachlorobiphenyl
total 14C
tetrachlorobiphenyl (Rf 0.48*)
Unknown I (Rf 0.23)
Unknown II (Rf 0.04)
Polar (Rf 0.0)
Unextractable
III. 2, 5,2', 4', 5'-
pentachlorobiphenyl
total 14C
pentachlorobiphenyl (Rf 0.55*)
Unknown I (Rf 0.46)
Unknown II (Rf 0.39)
Unknown III (Rf 0.21)
Unknown IV (Rf 0.04)
Unknown V (Rf 0.02)
H2°
0.02065
0.00120
0.00005
0.00155
0.01225
0.00560
0 . 04340
0.00985
--
0.00020
0.00015
0.00030
0.00385
Oedogonium
(alga)
23.6845
21.5975
0.3220
0.1030
0.3275
1.3345
62.4660
53.8440
0.6850
0.5080
0.1425
--
0.2570
Physa
(snail)
53.7465
47.3275
0.7560
0.4360
3.9850
1.2420
633.0165
587.3545
8.6210
2.2490
1.9365
0.5000
7.4965
Culex
(mosquito)
14.5335
12.6745
0.1070
--
0.9670
0.7850
181.4565
170.8480
2.4070
1.3195
1.0520
--
_ _
Gambusia
(fish)
15.5685
14.2360
0.0890
--
0 . 8545
0.3900
127.6945
119.7060
2.5380
0.5810
0.3285
--
0.7450

-------
                                           Table 38  (con't.)
Polar (Rf 0.0)
Unextractable
H2°
0.02055
0.00850
Oedogonium
(alga)
1.6265
5.4330
Physa
(snail)
16.5550
8.3040
Culex
(mosquito)
2.6745
3.1555
Gambusia
(fish)
2.3610
1.4350
*TLC with hexane (Skelly solve B b.p. 60-68°C)

-------
rats showed no adverse effects.  The mode of action  of  this   compound
is not clear as early work by Kittleson in 1952 suggests  that the
activity of this compound was related to the presence of  the  trichloro-
methyl group.  Later (Rich, 1960), the fungicidal activity was  ascribed
to-tfte production of thiophosgene after the displacement  of the
trichloromethyl group by a mercaptan group.

There appears to be some information about the environmental  stability
of captan.  The water solubility is reported to be less than  10 ppm and
undergoes hydrolysis in water of pH 7.0.  In addition,  the trichloro-
methyl sulfur moiety reacts readily with free thiol groups in
biological systems (Lukens, 1969).  The degradation in  soil appears to
proceed via hydrolysis with soil microorganisms playing a significant
role (Lukens, 1969).  The half-life in a moist, silt soil is  about 3-4
days and much longer in an air-dried soil (Burchfield,  1959) .   The
structures of the metabolites in soil are not completely known,
although the existence of carbonyl sulfide as a product has been
demonstrated (Somers e_t a±., 1967).

The behavior of captan in this terrestrial-aquatic model ecosystem
corroborates the previous data obtained from field experiments  as none
of the organisms after 33 days contained residues of this fungicide
(Table 39).  In view of the alkaline pH of the aqueous  portion  of the
system, the captan undoubtedly under went hydrolysis as soon  as it came
in contact with the water.  There were small amounts of unidentifiable
residues in the snail, fish and algae, but no captan, which had an Rf
of 0.3, was isolated from the organisms.  The interesting aspect of the
metabolites isolated from the snail was that they were much less polar
than captan.  Since the label was located on the trichloromethyl moiety,
perhaps the three spots are the result of a disulfide exchange with
captan and another mercaptan.

The water portion of the model ecosystem (Table 40) does not  contain
any traces of captan nor any of the nonpolar metabolites found  in
either the algae or snail.  Again as demonstrated for the organisms,
captan does not persist in the water as there were only metabolites which
had Rf values less than 0.35.  It appears that continued use  of captan
will not have any serious environmental impact as it does not persist
in the water of this 33-day model ecosystem,  nor does it accumulate in
the fish which is the upper member of the food chain.  The probable
reason for the nonpersistence of this fungicide in this model ecosystem
is the extremely labile trichloromethyl sulfur-nitrogen bond which can
be split either by hydrolysis or reaction with mercaptan groups in
biological systems.

                            Hexachlorophene

The final compound to be examined in the model ecosystem is the
bacteriostat, hexachlorophene.  The use of this antibacterial agent
has been restricted because of the reported poisonings in France from
baby powder that contained this compound.   Almost nothing is known about
                                 67

-------
CX5
                                                     Table 39

                                 Concentrations (ppm) of Captan and metabolites
                                        in organisms in a model ecosystem
Compound
iV
II
III
IV
V
VI
VII
VIII
IX
X
XI
XII
XIII
XIV
XV
R£a
0.93
0.85
0.81
0.79
0.68
0.39
0.35
0.33
0.26
0.25
0.18
0.14
0.10
0.053
0.00
Algae Daphnia Fish
0.0278
0.0077
__
0.0166
0.0492
0.0105
__
0.0142
__
0.00608
__
__
' 0.0590
0.0159 — 0.00215
0.122 — 0.000861
Mosquito Snail
0.0592
0.0795
0.0679
__
__
__
__
__
__
--
--
_-
_-
__
0.00941

-------
                                                 Table 39 (con't.)                                   t



           Compound          Rf^'          Algae        Daphnia         Fish         Mosquito        Snail
      Extractable ^C         .           0.280         0.393         0.0522         0.0462         0.216



      Unextractable 14C                  0.967         0.338         0.0158         0.0584         0.0998



      Grand Total 1(^C                    1.247         0.731         0.0680         0.105          0.316
      a/
      —'  Silica Gel GF-254, petroleum ether-acetone, 4:1 by volume

      ]*.'  Roman numerals - unknown spots
VO

-------
                               Table 40

           Concentrations  (ppm) of Captan and metabolites
                    in water in a model ecosystem
Compound
i*/
II
III
IV
V
VI
VII
VIII
IX
X
XI
XII
XIII
XIV
XV
Extractable 14C
Unextractable ^C
Grand Total 14C
«**'
0.93
0.85
0.81
0.79
0.68
0.39
0.35
0.33
0.26
0.25
0.18
0.14
0.10
0.053
0.00



Unhydrolyzed Water
--
--
--
--
--
--
0.00000426
--
0.00000960
--
0.00000456
0.00000273
0.00000365
0.0000648
0.00000761
0.0000971
0.000846
0.000943
Hydro lyzed Water
--
--
--
--
--
--
--
--
--
0.00000893
--
0.00000812
--
0.0000243
0.0000277
0.0000691
0.000777
0.000846
£/ Silica Gel GF-254, petroleum ether-acetone, 4:1 by volume
_' Roman numerals - unknown spots
                                 70

-------
this bisphenol and its metabolic products,  though  studies  with rats
(Wit and Van Genderen, 1962) and cows  (St.  John  and  Lisk,  1972)  have
demonstrated that the majority of the  hexachlorophene  is excreted
unchanged in the urine and feces when  fed to these animals.  However,
no-attempt was made to identify any of the  metabolites.  While some
work has been carried out on the metabolism of this  compound in  animals,
nothing is known about the environmental fate of this  material except
that it is thought to persist in surface waters  for  long periods  of
time without undergoing substantial degradation  (Bandt and Nehring,
1962).  Recently, hexachlorophene (HCP) has been detected  in sewer
influents in Oregon at levels of 20-31 ppb  and in  river water  upstream
from the city of Corvallis, Oregon, at 0.01-0.10 ppb.  Sewer treatment
of the influent removed about 60-70% of the HCP  (Buhler et: al^. ,  1973).
In view of the mammalian toxicity of hexachlorophene (Nakaue et al.,
1973) and the dearth of environmental degradation  information, it is
imperative to carry out some preliminary studies on  the fate of HCP  in
this model ecosystem.

The data for the fate of HCP in the organisms and water of this model
ecosystem are contained in Tables 41 and 42.  The  first aspect noticed
is the absence of Daphnia and mosquito larvae from the table.  This
indicates that these organians did not contain sufficient radioactivity
to be chromatographed.  Three of the organisms,  algae, 1.99 ppm;  fish,
0.37 ppm; and snail, 1.31 ppm contained identifiable amounts of
hexachlorophene along with several other uncharacterized metabolites.
Hexachlorophene represented about 42.8% of  the total radioactivity in
the algae, 18.3% in the fish and 17.4% in the snail.  The  rest of the
radioactivity in these organisms was distributed among ten uncharac-
terized metabolites.  While there were a substantial number of
metabolites in the three organisms, the unextractable radioactivity was
highest for the snail, 38%, intermediate for the algae, 25% and lowest
for the fish, 5.5%.

The water portion of the model ecosystem has small amounts of HCP,
0.00134 ppm, as well as numerous uncharacterized metabolites.  If the
figure for the hexachlorophene concentration in  the water, 0.00134 ppm,
is divided into the concentrations of HCP in the various organisms,
concentration factors for the algae, l,500x; fish, 278x; and snail,
970x are derived.  It can be concluded that the uptake of hexachloro-
phene by the fish is similar to that found for parathion,  which had a
concentration factor of about 335x.   Finally, the unextractable radio-
activity in the water amounts to about 36%, which indicates that
hexachlorophene is not extensively degraded to polar, unextractable
metabolites either by the organisms or chemical factors, such as
hydrolysis or photolysis.  The inert nature of hexachlorophene  is
related to its polar nature resultant from the two hydroxyl groups and
the highly substituted aromatic ring with chlorine and a methylene
bridge.  Higher chlorinated phenols such as 2,4,5-trichlorophenol and
2,3,4,6-tetrachlorophenol require about 10 weeks  to undergo ring
cleavage in soil (Alexander, 1972).   The 2,3,4,6-tetrachlorophenol is
                                 71

-------
precisely the same arrangement of substituents of hexachlorophene,
except the two position of HCP is filled by a methylene bridge instead
of a halogen.
                                  72

-------
                                Table 41

             Concentrations (ppm) of hexachlorophene and
            metabolites in organisms in a model ecosystem
Compound
hexachlorophene
£/
II
III
IV
V
VI
VII
VIII
IX
Extractable 14C

Unextractable 14C
Grand Total 14C
RfS/
0.89
0.82
0.75
0.70
0.65
0.53
0.23
0.14
0.07
0.00




Algae
1.994
0.608
--
--
--
0.563
0.0451
0.259
--
--
3.469
«
1.195
4.664
Fish
0.371
1.233
--
--
--
--
0.151
0.0261
0.0810
--
1.861

0.109
1.970
Snail
1.309
--
--
--
2.182
--
0.638
--
--
0.638
4.767

2.893
7.650
y Silica Gel GF-254, benzene-methanol-acetic acid, 45:8:4 by volume
—' Roman numerals - unknown spots
                                 73

-------
                               Table 42

               Concentrations  (ppm)  of hexachlorophene
            and metabolites  in water in a model ecosystem
     Compound
Unhydrolyzed Water
Hydrolyzed Water
hexachlorophene
£/
II
III
IV
V
VI
VII
VIII
IX
Extractable 14C
Unextractable ^C
Grand Total 14C
0.89
0.82
0.75
0.70
0.65
0.53
0.23
0.14
0.07
0.00



0.000542
--
0.00120
--
0.00183
0.000132
0.0000696
--
--
0.00000794
0.00378
0.03080
0.03458
0.000798
0.000444
0.00130
0.00374
0.00609
0.000536
0.00384
0.000542
--
0.000238
0.01753
0.00556
0.02309
—  Silica Gel GF-254, benzene-methanol-acetic acid, 45:8:4 by volume
b/ Roman numerals - unknown spots
                                  74

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                                Table 43

         Accumulation  factors  for  fish  and  snail  for  compounds
                    examined in the model ecosystem*
Insecticides

BuxR
SevinR
carbofuran
dieldrin
OrtheneR
parathion
lindane and Aroclor 5460R

Herbicides

alachlor
propachlor
BladexR
BentazonR
dicamba
2,4-D
pyrazon
TrifluralinR (sorghum treatment)
TrifluralinR (sand treafment)

Others

2,5,2'-trichlorobiphenyl
2,5,2l}5'-tetrachlorobiphenyl
2,4,5,2',5'-pentachlorobiphenyl
di-n-octyl phthalate
hexachlorophene
captan
BanomiteR
 Fish
 Snail
   335
 2,110
    452
   930
 4,200
 6,400
11,863
12,153
 9,400
   278
 17,700
153,000
  5,795
 39,439
 59,629
 13,600
    970
*Accumulation factor equal concentration of chemical (ppm) in the
   organism/concentration of chemical (ppm) in the water
                                 75

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                            SECTION VI

                            REFERENCES

Abernathy, John R., and L. M. Wax.  Weed Sci.  ^1:224,  1973.

Alexander, M.  In:  Environmental Toxicology of Pesticides,
  Matsumura, Fumio, G. Mallory Boush, and  Tomomasa Mi sato  (eds.).
  New York, Academic Press, 1972.  p. 365.

Anon.  Chem. Eng. News.  51 (23):13, 1973.

Anon.  Herbicide Handbook. Geneva, New York, The  W.  F.  Humphrey
  Press, Inc., 1970.  p. 5.

Bandt, H. J., and D. Nehring.  Z. Fisch.   10:543, 1962.

Beynon, K. I., G. Stoydin, and A. N. Wright.   Pest.  Sci.  _3:379,
  1972.

Broadhurst, N. A.,  M. L. Montgomery, and V. H. Freed.   J. Agr.  Food
  Chem.  _14:585,  1966.

Buhler, Donald R.,  M. E. Rasmusson, and H.  S.  Nakaue.   Environ.
  Sci. Technol.   7_:929,  1973.

Burchfield, H. P.   Contrib. Boyce Thompson Inst.  20:205,  1959.

Chevron Chemical  Company.  Technical Bulletin.   1972.

Childers, W. F.,  and W.  N. Bruce.  Unpublished data.  1973.

Dhillion, Nirmal  S. Dist. Abstra. Int. B.  _31:7136, 1971.

Frank, Richard.   Dist. Abstr.  Int. B.  _32:4444,  1972.

Friesen,  H.  A.  Weeds.   J.3:30,  1965.

Fukuto, T.  Roy, and James  J.  Sims.   In:   Pesticides  in the Environment,
  White-Stevens,  Robert  (ed.).   New  York,  Marcel Dekker,  Inc.,  1971.
  p. 145.

Grover, P.  L.,  and P.  Sims.   Biochem.  J.   96:521, 1965.

Gunther,  F.  A., W.  Westlake,  and P.  S.  Jaglan.  Residue Reviews.
   20:1,  1968.

Hargrove,  R. S.,  and M.  G.  Merkle.   Weed  Sci.   1.9:652, 1971.

Heath,  D.  F.,  and M.  Vanderkar.   Brit.  J.  Ind. Med.   21:269,  1964.

Hendersen,  C.,  A. Inglis,  and W. L.  Johnson.   Pest.  Monit.  J.
   5:1,  1971.


                                 76

-------
Herbicide Handbook.  Geneva, New York, The W. F. Humphrey  Press,
  Inc., 1970.  p. 136.

Horstein, I., and W. N. Sullivan.  J. Econ. Ent.  46:937,  1953.

Hurtig, Henry.  In:  Environmental Toxicology of Pesticides,
  Matsumura, Fumio, G. Mallory Boush, and Tomomasa Misato  (eds.).
  New York, Academic Press, 1972.  p. 257-

Hutson, D. H., M. H. Hoadley, M. H. Griffiths, and C. Donninger.
  J. Agr. Food Chem.  IS-.507, 1970.

Kaugars, Girts, Edwin G. Gemrich, and Victor L. Rizzo.  J. Agr.
  Food Chem.  21:647, 1973.

Kittleson, A.  Science.  115:84, 1952.

Knaak, James B.  Bull. Wld. Hlth. Org.  44:121, 1971.

Krieger, R. I., and P. W. Lee.  Arch. Environ. Cont. Tox.  J.:112,
  1973.

Ruhlman, D. E.  In:  Twenty-fifth Illinois Custom Spray Operators
  Training School, 1973.  p. 149.

Kuhlman, D. E.., and T. A. Cooley.  In:  Twenty-fifth Illinois Custom
  Spray Operators Training School, 1973.  p. 170.

Lamoreux, G. L., and F. S. Tanaka.  J. Agr. Food Chem.  JL9:346, 1971.

Lichtenstein, E. P., K. R. Schultz, T. W. Fuhremann, and T. T. Liang.
  J. Econ. Ent.  62:761, 1969.

Loos, M. A.  In:  Degradation of Herbicides, Kearney, P. C., and
  D. D. Kaufman (eds.).  New York, Marcel Dekker, Inc., 1969.  p. 1.

Lukens, R. J.  In:  Fungicides, Volume 2, Torgeson, D. E.  (ed.).
  New York, Academic Press, 1969.  p. 395.

Mayer, Foster L., Jun., David L. Stalling, and James L. Johnson.
  Nature.  238:411, 1972.

Metcalf, Robert L.  Essays in Toxicology.  5_:1, 1974.

Metcalf, Robert L., Inder P. Kapoor, Po-Yung Lu, Carter K. Schuth,
  and Patricia Sherman.  Environ. Hlth. Perspect.  p. 35, 1973.

Metcalf, Robert L., Gary M. Booth, Carter K. Schuth, Dale J. Hansen,
  and Po-Yung Lu.  Environ. Hlth. Perspect.  p. 29, 1973.

Metcalf, Robert L.  Bull. Wld. Hlth. Org.  44:43, 1971.
                                77

-------
Metcalf, Robert L.  In:  Pesticides in  the Environment, White-
  Stevens, Robert  (ed.).  New York, Marcel Dekker, Inc.,  1971.   p.  1.

Metcalf, Robert L., G. K. Sangha,  and I. P. Kapoor.   Environ. Sci.
  Technol.  _5:709, 1971.

Miskus, R. P., T.  L. Andrews, and  M. L. Look.   J. Agr. Food  Chem.
  17:842, 1969.

Moon, M. W., E. G. Gemrich,  and G. Kaugars.  J. Agr.  Food Chem.
  ^0:888, 1972.

Nakaue, H.  S., F.  N. Dost, and D.  R. Buhler.   Toxicol. Appl.  Pharm.
  24:239, 1973.

Parka,  S. J.,  and  H.-M.  Worth.  Proc. Southern Weed  Conf.  18:469,
  1965.

Peakall, David B.  Residue Reviews.  44:1,  1972.

Probst, G.  W., and J.  B. Tepe.  In:  Degradation  of  Herbicides,
  Kearney,  P.  C.,  and  D. D.  Kaufman  (eds.).  New  York, Marcel Dekker,
  Inc., 1969.  p.  255.

Rich,  S.  In:  Plant Pathology, Horsfall,  Jr., and A. Dimond (eds.).
  New  York,  Academic Press,  1960.  p. 588.

Riesbrough, R. W.  In:  Chemical Fallout,  Berg, G.  G., and M. W. Miller
   (eds.).   Springfield,  C.  C.  Thomas,  1969.  p. 5.

 Saha,  J. G.  J.  Econ.  Ent.   63:670,  1970.

 Sanders, Herman  0.,  Foster L.  Mayer, and David F. Walsh.   Environ.
  Research.  j>:84, 1973.

 Schultz, D. P.   J. Agr.  Food Chem.  21.: 186,  1963.

 Sechriest,  R. W.,  and D. W.  Sherrod.   In:   Twenty-fifth  Illinois Custom
   Spray Operators Training School, 1973.   p.  177.

 St. John,  Leigh E.,  and Donald J.  Lisk.  J.  Agr.  Food Chem.   20:289,  1972.

 Somers, E., D. V.  Richmond,  and J. A.  Pickard. Nature.   215:5097,  1967.

 Stephenson, G. R., and Stanley K.  Ries.  Weed  Sci.   JL7:327,  1969.

 Sutherland, G. L., J.  W. Cook,  and R.  L.  Baron.  J.  Assoc. Off.  Analyt.
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                                 78

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Tsao, Ching-Hsi, W. N. Sullivan, and Irwin Horstein.  J. Econ. Ent.
  46:882, 1953.

Tye, Russell, and David Engel.  J. Agr. Food Chem.  j.5:837, 1967.

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Wheeler, W. B.  J. Agr. Food Chem.  15:227, 231, 1967.

Wingo, C. W.  Res. Bull. Missouri Agr. Exptl. Sta.  914:27, 1966.

Wit, J. G., and H. Van Genderen.  Acta Physiol. Pharmacol. Neerlandica.
  11:123, 1962.
                                79

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                            SECTION VII

                             PUBLICATIONS

Booth, Gary M., John Connor, R. A. Metcalf and J. R. Larsen.  1973.
  A comparative study of the effects of selective inhibitors on
  esterase isozymes from the mosquito Anopheles punctipennis.  Comp.
  Biochem. Physiol., 44B:  1185-1195.

Sanborn, James R. and Ching-Chieh Yu.  1973.  The fate of dieldrin in
  a model ecosystem.  Bull. Environ. Contain. Toxicol., 10, 340.

Cho, P., R. Davenport, G. S. Whitt, G. M. Booth and J. R. Larsen.  1972.
  Electrophoretic and histochemical analysis of esterase synthesis in
  the ovariole of the milkweed bug Oncopeltus fasciatus  (Dall.).
  Wilhelm Roux1 Archiv., 170: 209-220.

Booth, Gary M. and Robert L. Metcalf.  1972.  The histochemical fate
  of paraoxon  in the cockroach (Periplaneta americana) and honey bee
  (Apis mellifera) brain.  Israel Journal of Entomology VII: 143-156.

Bruce, W. N.   1972.  Separation and identification of carbpfuran, its
  metabolites, and conjugates found in fish exposed to ring  l^C-labeled
  carbofuran using ITLC  silica gel strips.  J. Econ. Entomol. 65(6);
  1738-1740.

Yu, Ching-Chieh, Robert  L. Metcalf and Gary M  Booth.  1972.  Inhibi-
  tion of acetylcholinesterase from mammals and insects by carbofuran
  and its related compounds and their toxicities toward  these animals.
  Agr. and Food Chem., 20(5): 923-926,

                           In Preparation

Yu, Ching-Chieh and James R. Sanborn.  The fate of parathion in a
  model ecosystem.  To be submitted to Bull. Environ. Contam. Toxicol.

Sanborn, James R., Robert L. Metcalf and William F. Childers.  The
  uptake of three PCB's, DDT and DDE by the green sun fish Lepomis
  cyanellus Raf.  To be  submitted to Bull. Environ. Contam.  Toxicol.

Sanborn, James R., Robert L. Metcalf, Ching-Chieh Yu and Po-Yung Lu.
  Plasticizers  in  the environment.  The fate of di-n-octyl phthalate
   (DOP) in  two model ecosystems and uptake and metabolism of DOP by
  aquatic organisms.  To be submitted to Arch. Environ.  Contam. Toxicol.

Sanborn,  James R.  and Ching-Chieh Yu.  The fate of TreflaiiB) in a model
  ecosystem:   A  comparison of  two methods of treatment.  To  be submitted
  to  Bull. Environ. Contam. Toxicol.
                                  80

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Metcalf, Robert L., James R. Sanborn, Po-Yung Lu and Donald E. Nye.
  Laboratory model ecosystem studies of degradation and fate of radio-
  labeled tri-, tetra-, and pentachlorobiphenyl compared with DDE.   To
  be submitted to Arch. Environ. Cont. Toxicol.
                               81

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   CHEMICAL NOMENCLATURE

       Compound
Sevii®

car bo fur an


dieldrin



lindane

Orthene®

parathion

alachlor


propachlor

BladeJ)


Bentazori®


dicamba

2,4-D

Pyrazon®

TrifluralinR


Banomite®


trichlorobiphenyl

tetrachlorobiphenyl

pentachlorobiphenyl
    SECTION  VIII

FOR COMPOUNDS EXAMINED IN A MODEL ECOSYSTEM

                      Name

3:1 mixture of m-(l-ethylpropyl)phenyl N-
  methylcarbamate and m-(l-methylbutyl)phenyl N-
  methylcarbamate

1-napthyl N-methylcarbamate

2,2-dimethyl-2,3-dihydrobenzofuranyl 7-N-
  methylcarbamate

1,2,3,4,10,10-hexachloro-6,7-epoxy-l,4,4a,5,
  6,7,8a-octahy_
-------
      Compound                                  Name

DOP                      di-n-octyl phthalate

captan                   N-trichloromethylthio-4-cyclohexene-l,2-
                           dicarboximide

hexachlorophene          2,2-methylene-bis (3,4,6-trichlorophenol)
                                 83

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 SELECTED WATER
 RESOURCES ABSTRACTS
 INPUT TRANSACTION FORM
         rt No.
 4.  Title
                FATE QF SELECT PESTICIDES IN THE
           AQUATIC ENVIRONMENT
 7. Author(3) sanborn,  James  R.
 9. Organization  Illinois Natural History Survey and
               Board of Trustees, University of Illinois,
               Urbana,  Illinois


J2. Sponsoring Ore*™* ion   ENVIRONMENTAL PROTECTION AGENCY
                                                                  3.
                                                                          ioz No,
                        w
                        5.  l
                        6.   '
                        8.  Performing Otg*a
                           Report No,

                       if).  Project No.

                        R-800736
                        i.  Contrac':jGrantNo
                        Grant R-800736

                       13.  Type of Repor: and
                           Period Covered Final
 15.
 16.  Abstract
In this  study  17  organic pesticides and five industrial  chemicals were examined in a
terrestrial-aquatic model ecosystem in an effort to determine their persistence and
accumulation by the organisms of this system.  Several classes of pesticides are
represented as one or more insecticides, herbicides, miticides or plasticizers were
investigated in this system.   The use of this system for examining uptake and per-
sistence of widely used agricultural chemicals provides  the  necessary data for compari-
son of field data to provide a framework which can be used to assess the potential
environmental  impact of new pesticides before they are given a recommendation for
generalized use.

The data obtained from this work suggest that this model ecosystem is useful for the
determination  of  the uptake and persistence of pesticides by the  organisms-.  In general,
it was found that most chemicals, with the exception of  the  persistent soil insecticide,
dieldrin, underwent extensive degradation under the experimental  conditions of the
system.  Dieldrin was exceptional in its behavior in that >96% of the radioactivity
isolated from  the organisms was  unchanged dieldrin, clearly  indicating the extreme
inertness of this chlorinated hydrocarbon to undergo biological or  chemical
modification.
 17a. Descriptors
*Ecosysterns, Trophic  levels,  *Pollutants,  *Pesticide residues, *Biodegradation,
Ecological distribution
 l"b. Identi inrs
Model ecosystem, Insecticides,  Herbicides,  Plasticizers, Polychlorinated Biphenyls,
Bux, Carbaryl, Carbofuran,  Dieldrin,  Lindane, Orthene, Parathion, Alachlor,  Propachlor,
Bladex, Bentazon, Dicamba,  Pyrazon,  2,4-Dichlorophenoxyacetic Acid, Trifluralin,  Banomit
DOP, Chlorinated Biphenyls,  Captan,  Hexachlorophene, Fish, Snail, Algae


                      05A, 05B
                        19.  S1  -\rity C' ss.
                                    di'i.
21. A«j. of
   Pc.Se-

2t. Price
Send To:
WATER RESOURCES SCIENTIFIC INFORMATION CENTER
U.S. DEPARTMENT OF THE INTERIOR
WASHINGTON, D C 2O24O
          James R. Sanborn
     Illinois Natural History Survey
                      •if U.S. GOVERNMENT PRINTING OFFICE: 1975-697-819/77 REGION 10

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