&EPA
            United States
            Environmental Protection
            Agency
            Municipal Environmental Research  EPA-600 2-80124
            Laboratory         August 1980
            Cincinnati OH 45268
            Research and Development
Adsorption,
Movement, and
Biological
Degradation of Large
Concentrations of
Selected Pesticides
in Soils

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                RESEARCH REPORTING SERIES

Research reports of the Office of Research and Development, U.S. Environmental
Protection Agency, have been grouped into nine series. These nine broad cate-
gories were established to facilitate further deveJopment and application of en-
vironmental technology. Elimination of traditional grouping was  consciously
planned to foster technology transfer and a maximum interface in related fields.
The nine series are:

      1.  Environmental Health  Effects Research
      2.  Environmental Protection Technology
      3.  Ecological Research
      4.  Environmental Monitoring
      5.  Socioeconomic Environmental Studies
      6.  Scientific and Technical Assessment Reports (STAR)
      7.  Interagency Energy-Environment Research and Development
      8.  "Special" Reports
      9.  Miscellaneous Reports

This report has  been assigned  to the  ENVIRONMENTAL PROTECTION TECH-
NOLOGY series. This series describes research performed to develop and dem-
onstrate instrumentation, equipment, and methodology to repair or prevent en-
vironmental degradation from point and non-point sources of pollution. This work
provides the new or improved technology required for the control and treatment
of pollution sources to meet environmental quality standards.
This document is available to the public through the National Technical Informa-
tion Service, Springfield, Virginia  22161.

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                                    EPA-600/2-80-124
                                    August 1980
   ADSORPTION, MOVEMENT, AND BIOLOGICAL
   DEGRADATION OF LARGE CONCENTRATIONS
          OF SELECTED PESTICIDES
                 IN SOILS
                    by
              J. M. Davidson
               P. S. C. Rao
                 L. T. Ou
               W. B. Wheeler
              D. F. Rothwell
          Soil Science Department
           University of Florida
           Gainesville, Florida
            Grant No. R-803849
              Proj ect Officer,

               M. H. Roulier
Solid and Hazardous Waste Research Division
Municipal Environmental Research Laboratory
          Cincinnati, Ohio  45268
MUNICIPAL ENVIRONMENTAL RESEARCH LABORATORY
    OFFICE OF RESEARCH AND DEVELOPMENT
   U. S. ENVIRONMENTAL PROTECTION AGENCY
          CINCINNATI, OHIO  45268

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                                 DISCLAIMER


     This report has been reviewed by the Municipal Environmental Research
Laboratory, U. S. Environmental Protection Agency, and approved for publi-
cation.  Approval does not signify that the contents necessarily reflect
the views and policies of the U. S. Environmental Protection Agency, nor
does mention of trade names or commercial products constitute endorsement
or recommendation for use.
                                      11

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                                  FOREWORD
     the Environmental Protection Agency was created because of increasing
public and government concern about the dangers of pollution to the health
and welfare of the American people.  Noxious air, foul water, and spoiled
land are tragic testimony to the deterioration of our natural environment.
The complexity of that environment and the interplay between its components
require a concentrated and intergrated attack on the problem.

     Research and development is that necessary first step in problem solu-
tion and it involves defining the problem, measuring its impact, and search-
ing for solutions.  The Municipal Environmental Research Laboratory develops
new and improved technology and systems for prevention, treatment, and man-
agement of wastewater and solid and hazardous waste pollutant discharges
from municipal and community sources, for the preservation and treatment of
public drinking water supplies, and to minimize the adverse economic,
social, health, and aesthetic effects of pollution.  This publication is
one of the products of that research; a most vital communication link
between the researcher and the user community.

     This report presents the results of a study on the behavior of selected
pesticides in soils when the pesticide is present at large concentrations.
The work was conducted because of uncertainty whether the existing low-
concentration agricultural data base could be extrapolated to predict the
behavior of large concentrations of pesticides in soils at disposal sites.
                                          Francis T. Mayo, Director
                                          Municipal Environmental Research
                                          Laboratory
                                     111

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                                  ABSTRACT


     Because of the importance of soil in biologically reducing the quantity
 and retarding the rate of pollutant movement into groundwater, this labora-
 tory study was initiated to evaluate the adsorption, mobility, and degrada-
 tion of large concentrations of the pesticides atrazine, methyl parathion,
 terbacil, trifluralin, and 2,4-D in soils representing four major soil orders
 in the  United States.

     Equilibrium adsorption isotherms of the non-linear Freundlich type were
 obtained for all pesticides and the four soils.  Pesticide solution concen-
 trations used in the study ranged from zero to the aqueous solubility limit
 of each pesticide.  The mobility of each pesticide increased as the concen-
 tration of the pesticide in the soil solution phase increased.  These results
 were in agreement with the equilibrium adsorption isotherm data.  Pesticide
 degradation rates and soil microbial populations generally declined as the
 pesticide concentration in the soil increased; however, some soils were able
 to degrade a pesticide at all concentrations studied, while others remained
 essentially steril throughout the incubation period (50 to 80 days).  As
 shown by measurements of 14C02 evolution, total CO-2 evolution was not always
 a  good  indication of pesticide degradation.  Several pesticide metabolites
 were formed and identified in various soil-pesticide systems.  The quantities
 of trifluralin and atrazine "bound" to the soil at the end of the incubation
 period  were measured and in some cases appeared to be related to types of
 metabolites formed during biological degradation.

     The observed increase in pesticide mobility for large pesticide concen-
 trations in the soil invalidates, in many cases, the usefulness of the
 existing low concentration data base for designing pesticide waste disposal
 sites.   Owing to the increased mobility and lower microbial decompostion
 rates of many pesticides when introduced into soils at waste disposal con-
 centrations, the potential for groundwater contamination is increased sig-
nitifantly.  The data presented in this report should be given consideration
when designing waste disposal sites for pesticides.  The low concentration
 data base for pesticide mobility may be reasonable when considering pesticides
with low aqueous solubilities; however, this should not be accepted without
verification.

     This report was submitted in fulfillment of Grant No. R-803849 by the
University of Florida, Gainesville, under the partial sponsorship of the U. S.
Environmental Protection Agency.  This report covers the period August 1, 1975
to October 31, 1977, and work was completed as of October 31, 1977.
                                      IV

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                                  CONTENTS
Foreword	  iii
Abstract	   iv
Figures	   vi
Tables	   ix
Acknowledgements	   xi

     1.  Introduction	    1
     2.  Conclusions	    3
     3.  Recommendations	    5
     4.  Materials and Methods	    6
               Soils	    6
               Pesticides	    6
               Adsorption Isotherms	    9
               Pesticide Displacement through Soils	   10
               Pesticide Degradation and Soil Respiration	   11
               Soil Microbial Populations	   11
               Metabolite Identification	   12
     5.  Results and Discussion	   15
               Adsorption and Mobility	   15
                    Adsorption Experiments	   15
                    Column Displacement Experiments	   26
                    Simulation of Pesticide
                    Mobility in Soils	   32
                    Infiltration Experiments	   35
               Microbial Acitvity and Degradation	   42
                    Atrazine	   42
                    Methyl Parathion	   45
                    Trif luralin	   58
                    2,4-D	,	   61
               Metabolites	   90
                    Atrazine;	   90
                    Trif luralin	   90
                    2,4-D	   99
     6.  Implications of Project Results With Regards to
         Pesticide Disposal	  101

References	  103

Appendix:  List of Publications from this project	  109

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                                   FIGURES
Figure

   1   Adsorption isotherm for 2,4-D amine and Webster soil.  Data
         obtained using soil:solution ratios of  1:2,  1:5 and  1:10..    17

   2   Adsorption isotherm for 2,4-D amine and Webster soil.  2,4-D
         was applied to soil in distilled water  or 0.01N CaCl2	    18

   3   Adsorption isotherm for 2,4-D amine and Cecil  soil.  2,4-D
         was applied to soil in distilled water  or 0.01N CaCl2	    19

   4   Adsorption isotherms for atrazine and Webster, Cecil,  Glen-
         dale and Eustis soils.  Freundlich constants (K and  N) for
         each isotherm, determined by least-squares fit to the data,
         are also shown	    21

   5   Adsorption isotherms for methyl parathion and Webster, Cecil,
         Glendale and Eustis soils.  Freundlich  constants (K  and N)
         for each isotherm, determined by least-squares fit to the
         data, are also shown	    22

   6   Adsorption isotherms for terbacil and Webster, Cecil,  Glendale
         and Eustis soils.  Freundlich constants (K and N) for each
         isotherm, determined by least-squares fit to the data, are
         also shown	    23

   7   Adsorption isotherms for trifluralin and Webster, Cecil, Glen-
         dale and Eustis soils.  Freundlich constants (K and N) for
         each isotherm, determined by least-squares fit to data, are
         also shown	    24

   8   Adsorption isotherms for 2,4-D amine and Webster and Cecil
         soils.  Freundlich constants (K and N)  for each isotherm,
         determined by least-squares fit to data, are also shown...    25

   9   Effluent breakthrough curves for 2,4-D amine (C  = 50  and
         5,000 yg/ml) and for tritiated water displacement through
         Webster soil column	    28

  10   Effluent breakthrough curves for 2,4-D amine (C  = 50  and
         5,000 yg/ml) and for tritiated water displacement through
         Cecil soil column	    29
                                     VI

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Figure                                                                 Page

  11   Effluent breakthrough curves for 2,4-D arnine  (C   =  50  and
         5,000 yg/ml) and for tritiated water displacement through
         Eustis soil column	     30

  12   Effluent breakthrough curves for atrazine  (C   =  5 and  50
         yg/ml) and for tritiated water displacement  through
         Eustis soil column	     31

  13   Measured and simulated breakthrough curves  for 2,4-D amine
         displacement through Cecil soil column.   Parameter values
         used to calculate the solid lines were obtained from a
         2-parameter fit, and those for dashed lines  were  estimated
         from a 4-parameter fit to C  =50 yg/ml data	     37

  14   Soil-water content (solid line) and 2,4-D,  terbacil and
         atrazine concentration distribution in Eustis  soil follow-
         ing infiltration of water to approximately 30  cm.  Soil was
         initially air dry and herbicide was in the top 1.5 cm seg-
         ment of the soil (2,000 yg/g of soil)	     40

  15   Soil-water content (solid lines) and terbacil  concentration
         distributions in Eustis, Cecil and Webster soils  following
         infiltration of water to approximately 30 cm.   Soils were
         initially air dry and herbicide was in the top 1.5 cm seg-
         ment of the soil (2,000 yg/g of soil)	     41

  16   CC>2 evolution rate from the Cecil soil receiving various
         concentrations of atrazine.  (A) Technical grade;  (B)
         Formulated atrazine	     43

  17   CC>2 evoltuion rate from the Web,ster soil receiving  various
         concentrations of atrazine.  (A) Technical grade;  (B)
         Formulated atrazine	,	     44

  18   C02 evolution rate from the Cecil soil receiving various
         concentrations of trifluralin.  (A) Technical  grade;  (B)
         Formulated trifluralin	     59

  19   C02 evolution rate from the Webster soil receiving  various
         concentrations of trifluralin.  (A) Technical  grade;  (B)
         Formulated trifluralin	     60

  20   C02 evolution rate from the Webster soil receiving  various
         concentrations of 2,4-D.  (A) Technical grade;  (B) Formu-
         lated 2,4-D	     71

  21   Percent 1IfC02 evolved from the Webster soil receiving  various
         concentrations of 2,4-D.  (A) Technical grade;  (B) Formu-
         lated 2,4-D	,	     73
                                     vii

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Figure                                                                Page

  22   C02 evolution rate from the Cecil soil receiving various
         concentrations of 2,4-D.   (A)  Technical grade; (B)
         Formulated 2,4-D	     74

  23   Percent 14C02 evolved from the Cecil soil receiving various
         concentrations of 2,4-D.   (A)  Technical grade; (B)  Formu-
         lated 2,4-D	     75

  24   C02 evolution rate from the Terra Ceia soil receiving 5,000
         and 20,000 ppm of technical grade and formulated 2,4-D....     76

  25   Percent 14C02 evolved from the Terra Ceia soil receiving
         5,000 and 20,000 ppm of technical grade and formulated
         2,4-D	     77

  26   Percent of total 14C-activity bound to Webster and Cecil soil
         after receiving 10, 1,000 and 20,000 ppm of technical grade
         and formulated atrazine	     91

  27   Percent of total 14C-activity bound to Webster and Cecil soil
         after receiving 10, 1,000 and 20,000 ppm of technical grade
         and formulated trifluralin	     98

  28   Metabolic pathways for trifluralin metabolism.  Code numbers
         correspond with those in Tables 26 and 27	    100
                                    viii

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                                   TABLES
Table                                                                 Page

  1   Physical and chemical properties of the soils used in this
        study ....... * ..............................................     7

  2   Properties of pesticides used in this study ..................     8

  3   Solvents used for pesticide extraction from soil .............    12

  4   Thin layer chromatography systems ............................    14

  5   Freundlich constants calculated from equilibrium adsorption
        isotherms for various soil -pesticide combinations ..........    16

  6   Comparison of model parameter values estimated from the 2,4-D
        amine breakthrough data (C  = 50 yg/ml) for Cecil soil by
        varying either two or four°parameters in the nonlinear least-
        squares curve-fitting procedure ............................    36

  7   Physical data for infiltration experiments . . ^ ................    39

  8   Effect of technical grade and formulated atrazine on bacterial
        populations in Cecil and Webster soils . . . . : ................    46

  9   Effect of technical grade and fromulated atrazine on fungal
        populations in Cecil and Webster soils .....................    49

 10   Effect of technical grade and formulated atrazine on actino-
        mycete populations in Cecil and Webster soils ..............    52

 11   Total 0)2 evolution from methyl parathion treated Webster
        soil ............................. . .........................    55

 12   Total C02 evolution from methyl parathion treated Cecil
        soil [[[    56
 13   Percent of ^C -methyl parathion evolved as ll*C02 in Webster
        and Cecil soils ............................................    57

 14   Effect of technical grade and formulated trifluralin on

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Table                                                                 Page

 15   Effect of technical grade and formulated trifluralin on fungal
        populations in Cecil and Webster soils .....................     55

 16   Effect of technical grade and formulated trifluralin on actino-
        mycete populations in Cecil and Webster soils ..............     68

 17   Comparison of the percentage of 2,4-D degradation from the
        Webster soil derived from lkCQ2 and total C02 evolution during
        the second peak period and the total experimental period. . .     78

 18   Degradation rates for the three soils receiving 5,000 ppm of
        technical grade and formulated 2,4-D during exponential degra-
        dation period ..............................................     79

 19   2,4-D degradation in the Cecil soil receiving 5,000 ppm of the
        herbicide and various nutrient treatments during 60 days of
        incubation .................................................     81

 20   Soil pH of the Webster soil recieving technical grade and
        formulated 2,4-D ...........................................     82

 21   Soil pH of the Cecil soil receiving technical grade and
        formulated 2,4-D ...........................................     82

 22   Effect of 2,4-D on bacterial populations in Cecil soil .......     84

 23   Effect of 2,4-D on fungal populations in Cecil soil ..........     86

 24   Effect of 2,4-D on actinomycete populations in Cecil soil....     88

 25   Percentage of applied trifluralin mineralized and volatilized
        after 83 days of incubation ................................     93
 26   R  values for trifluralin and metabolite references
 27   Chromatographic and mass spectral parameters of products
        isolated from trifluralin treated soil .....................     95

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                              ACKNOWLEDGEMENTS
     The assistance of Mr. R. E. Jessup  (University of Florida] in develop-
ing numerical solutions to the conceptual mathematical models used to de-
scribe pesticide adsorption and mobility in soils is gratefully acknowl-
edged.  Also, the cooperation and overall project coordination by Dr. M. H.
Roulier (EPA, Municipal Environmental Research Laboratory, Cincinnati,
Ohio) as the Project Officer is appreciated.

     The project investigators wish to express their appreciation to the
Center for Environmental Programs in the Institute of Food and Agricultural
Sciences at the University of Florida for financial support during the
project.

     The following companies are acknowledged for providing pesticidal
materials used in this work:
          CIBA-Geigy for Atrazine (technical material, formulation,
             11+C-atrazine, authentic metabolites);
          Dupont for Terbacil (technical material, formulation,
             llfC-terbacil);
          Eli Lilly for Trefluralin (technical material, formula-
             tion, 14C-trifluralin, authentic metabolites);
          Kerr-McGee for Methyl Parathion (formulation);
          Monsanto for Methyl Parathion  (technical material);
          Thompson-Hayward for 2,4-D (formulation).
                                     XI

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                                  SECTION 1

                                INTRODUCTION
     Because of a continued increase in the number and quantity of pesticide
compounds being placed on the market, the safe disposal of surplus and/or
waste pesticide materials has become an acute problem (von Rumker, et al.,
1974).  Incineration, encapsulation, isolation in underground caves and
mines, chemical stabilization, land spreading and landfills are some of the
procedures being considered for the disposal of pesticides and other
hazardous wastes (Schomaker, 1976; von Everdingen and Freeze, 1971; Wilkin-
son, et al., 1978).  Of these methods, disposal by landfills and land
spreading appear to be more common and economical (Fields and Lindsey,
1975; Lindsey, et al., 1976).  Placing hazardous wastes in the land has
come under attack recently (Atkins, 1972; Rouston and Wildung, 1969) because
there is no guarantee that the hazardous chemicals disposed of in this
manner will not migrate from the disposal site to potable water supplies.

     In general, pesticide applications associated with agricultural pro-
duction have had very little adverse effect on the soil microbial activity
(Cole, 1976; Hubbell, et al., 1973; Jensen, 1962; Kaiser, et al., 1970;
Newman and Downing, 1958; Roslycky, 1977).  However, reports on soil micro-
bial activity where large concentrations were used have been contradictory
and inconclusive.  For example, Ou et al. (1978a) observed 2,4-D (2,4-di-
chlorophenoxyacetic acid) degradation at concentrations of 5,000 and 20,000
yg/g of soil (ppm) for one soil type and no degradation at the same concen-
trations for another soil.  Soil respiration and bacterial, fungal and
actinomycete populations were significantly reduced in the soil unable to
degrade 2,4-D.  They concluded that the physical and chemical properties of
the soil as well as the 2,4-D concentration were important factors in
governing microbial activity and pesticide degradation in soils receiving
large pesticide concentrations.

     Trifluralin (a,a,a-trifluro-2,6-dinitro-N, N-dipropyl-p-toluidine) and
atrazine (2-chloro-4-ethylamine-6-isopropylamino-s-triazine) are commonly
used herbicides.  Trifluralin applications of 3 kg/ha (1.4 ppm) have been
shown not to influence soil bacterial, fungal and actinomycete populations
significantly (Tyunaeva, 1974).  However, when 1.1 kg/ha (0.5 ppm) was
applied per year over a five year period, bacterial populations were in-
hibited while fungal and actinomycete populations were enhanced (Breazeale
and Camper, 1970).  When analytical grade trifluralin was incorporated into
the soil at concentrations of 5,000 ppm, C02 evolution and bacterial popu-
lations were inhibited while streptomycete populations were stimulated.
Stojanovic et al. (1972) has shown that formulated trifluralin stimulated
C02 evolution and streptomycete populations while inhibiting bacterial

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populations in soil. Atrazine was shown not to inhibit soil respiration at
concentrations associated with agricultural production (Kaiser, et al.,
1970; Eno, 1962). Cole  (1976) and Voets et al. (1974) have shown that soil
bacterial and fungal populations were not affected at rates below 4 kg/ha
(1.8 ppm).  However, Stojanovic et al. (1972) reported that atrazine iiihib-
ited soil respiration and bacterial populations at 5,000 ppm but had no
effect on fungal populations.

     A thorough understanding of the various processes that influence the
persistence, retention, and leaching of pesticides in soils is required to
develop technology for the selection and management of pesticide disposal
sites involving soils.  The fate of pesticides in soils when applied at
concentrations similar to those associated with agricultural practices has
been well-documented in several reviews (Bailey and White, 1970; Sanborn,
et al., 1977).  However, the direct extrapolation of this data base to
systems containing large pesticide concentrations, such as those occurring
at or below disposal sites, may not be feasible (Davidson, et al., 1976).

     Laboratory experiments were initiated to investigate the physical,
chemical and microbiological behavior of five pesticides in five soils when
the pesticide was present at large concentrations.  The objectives of the
study were to:  1) Measure and describe pesticide adsorption in selected
soil-water systems over a wide range of chemical concentrations (zero to
water solubility), 2) Measure mobility and distribution of pesticides in
selected soils, when applied or initially  present in soil at large concen-
trations, 3) Measure chemical and microbial degradation rate, and identify
metabolites produced in soil-pesticide systems receiving large pesticide
concentrations, and 4) Measure the influence of large pesticide concentra-
tions on soil microbial activity and respiration rate for selected soil-
pesticide systems.

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                                  SECTION 2

                                 CONCLUSIONS
     1.  Equilibrium adsorption isotherms for pesticides and soils consider-
ed in this study were nonlinear.  The Freundlich equation described the
full range of pesticide solution concentrations (zero to aqueous solubility
limit) studied.  The adsorption "sites" for the pesticide were apparently
never saturated in any soil-pesticide system investigated.

     2.  Pesticide mobility was inversely related to the pesticide concen-
tration in the soil solution phase when the adsorption isotherm was non-
linear (N < 1.0).  2,4-D at 5,000 yg/ml was nearly as mobile as the chloride
ion. The dependence of the mobility on pesticide solution concentration
could be predicted and described by the equilibrium adsorption isotherm.

     3.  14C02 evolution rate from a 14C-ring labeled pesticide was a good
indicator of pesticide degradation rate in soils containing large pesticide
concentrations.  Total C02 evolution, however, did not always represent the
actual degradation of a pesticide.  For this reason, the biological activity
of soils containing low pesticide concentrations may, in many cases, be
unrelated to the behavior and degradation of pesticides at large concentra-
tions in the soil.

     4.  Moderately persistent pesticides such as atrazine and trifluralin,
degrade slowly forming a number of metabolites when present in soils at
high and low concentrations.  These pesticides and metabolite products have
the potential to become "bound" to the soil matrix.  Bound residue was de-
fined in this study to be the 14C-labeled material remaining in the soil
after recommended extraction procedures had been employed.

     5.  The less persistent pesticides such as 2,4-D and methyl parathion
may or may not degrade when applied to a soil at high concentrations.
Factors that determine the degradation rate were pesticide concentration,
chemical formulation, nutrients in the soil, soil type, soil pH, temperature,
soil-water content, soil organic matter content, texture, and the presence
of a microbial population capable of degrading the pesticide.  If the
pesticide was degraded, the time required for degradation to begin was
longer for large concentrations (>50 ppm) than for low concentrations.  The
lag period was directly related to pesticide concentration.

     6.  Pesticide mobility and degradation were described using adsorption
and biological degradation parameters measured during this study.  Con-
ceptual or process based mathematical models were used to describe pesticide

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mobility.  The models were well suited for estimating  the concentration
distribution and mobility of pesticides associated with waste disposal
sites.

     7.  Bound residues were formed at all concentrations studied (10 to
20,000 ppm).  The percent of binding was higher for low concentrations;
however, the amount bound (atrazine or trifluralin) increased as the pesti-
cide concentration increased.  The toxicity and chemical nature of the
"bound residues" were not studied in detail.

     8.  At low pesticide concentrations (<50 ppm), microbial activity
(indicated by total C02 evolution, and bacterial, fungal and actinomycete
population) was generally not affected.  Microbial activity may be enhanced
or inhibited when a large amount of pesticide is applied to a soil.   When
degradation occured at large concentrations, microbial activity was enhanced.
Moreover, formulation chemicals may stimulate microbial activity by serving
as energy or nutrient sources.  If pesticide degradation does not occur,
microbial activity may be either enhanced or inhibited.

     9.  Based on the results of this study, groundwater contamination may
be a significant problem when highly soluble pesticides are placed in waste
disposal sites subject to considerable leaching.  The potential for ground-
water contamination, on the other hand, is not as great for pesticides with
low solubilities.

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                                  SECTION 3

                               RECOMMENDATIONS
     1.  Total C02 evolution or soil respiration should not be used as the
only measurement to estimate pesticide degradation in soils receiving or
containing large pesticide concentrations.

     2.  Because this study considered only systems involving the presence
of one pesticide, additional studies need to be conducted to establish the
degradation, adsorption, and movement of pesticides in systems containing
mixtures of several pesticides at high and low concentrations.  This type
of information is more relevant to actual pesticide disposal sites.

     3.  Identify procedures for stimulating pesticide degradation in waste
disposal sites involving soils.

     4.  Establish the major soil parameters and properties which determine
the potential for a given soil to degrade a high concentration of a given
pesticide.  Factors to consider are microbial population and species, soil
pH, organic matter content, cation exchange capacity, soil-water content,
temperature, etc.

     5.  Determine the mechanisms responsible for the formation of "bound"
residues. Evaluate stability and/or phytotoxicity of bound residues.

     6.  Develop   protocols for site evaluation and selection for pesticide
disposal using surface soils or landfills.  Protocols should consider
single as well as mixtures of pesticides.

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                                  SECTION 4

                            MATERIALS AND METHODS
SOILS
     Soils used in this study were selected on the basis of their geographic
and taxonomic representation of major soil orders in the United States.
State Conservationists for the Soil Conservation Service in selected regions
were asked to identify the major soil series in their area for a selected
soil order and to ship 735 kg (air-dry) of the Ap horizon of that soil.  A
soil profile description, sample site location, previous crop and management
history, and climatic conditions at the site were provided with each soil
series selected and studied.  Soils selected were: Webster silty clay loam
(Typic Haplaquolls) from Iowa, Cecil sandy loam (Typic Hapludults) from
Georgia, Glendale sandy clay loam (Typic Torrifluvents) from New Mexico,
Eustis fine sand (Typic Quartzipsamments), and Terra Ceia muck (Typic
Mediasaprista) from Florida.

     The soils were air-dried and sieved to pass a 2-mm screen prior to
being stored.  Selected physical and chemical properties of the mineral
soils are given in Table 1.  Terra Ceia muck is characterized by 81% organic
matter, 19% total mineral content, CEC of 350 meq/lOOg and pH of 6.4.

PESTICIDES

     The pesticides used in this study were selected based upon their
present and anticipated usage as well as their different chemical properties.
The production and use of herbicides has increased significantly in the
past few years and at the present time herbicides represent the largest
group of pesticides on the market.  Because of this market shift in pesticide
production, the herbicides were identified as a group of chemicals requiring
major attention.  The following description of each pesticide used in the
study illustrates the diversity in chemical properties and toxicity of the
selected compounds.  It was believed that this range in chemical properties
(Tabel 2) provided the necessary information needed to evaluate the problems
associated with introducing large pesticide concentrations into the soil
environment.
1)   Atrazine (Herbicide):  (2-chloro-4-ethylamino-6-isopropylamino-s^-
     triazine).Low solubility in water (Table 2), but highly soluble in
     chloroform, methanol, and ether.  Losses due to chemical and microbial
     degradation are significant.  Leaching from soils may be limited due
     to adsorption on certain soil constituents. Acute oral LD50 to rats
     3080 mg/kg.

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TABLE 1.  PHYSICAL AND CHEMICAL PROPERTIES OF .THE MINERAL SOILS USED IN THIS STUDY

PARTICLE
SOIL Sand
Webster 18.4
Cecil 65.8
Glendale 50.7
Eustis 93.8
SIZE FRACTION
Silt
45.3
19.5
16.4
3.0
(*)
Clay
38.3
14.7
22.9
3.2
pH (1:
Water
7.3
5.6
7.4
5.6
1 paste)
IN KC1
6.5
4.8
6.5
4.1
CEC
(meq/lOOg)
54.7
6.8
35.8
5.2
Organic C
(%)
3.87
0.90
0.50
0.56
Base
Saturation
(%3
91
31
90
10 _
Extractable
Acidity
(meq/lOOg)
5.15
4.68
3.74
4.68

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            TABLE 2.  PROPERTIES OF PESTICIDES USED IN THIS STUDY

Common
Name
Atrazine
Methyl
parathion
Terbicil
Trifluralin
2,4-D
2,4-D Amine
Molecular
Weight
215.7
263.2
216.7
335.3
221.0
266.1
Vapor
Pressure
(mmHg x 106)
1.4 (30° C)
9.5 (20° C)
0.48 (29.5° C)
199 (29.5° C)
n.d.*
0.001 (28° C)
Aqueous
Solubility
Cg/100 ml H20)
0.0033 (27° C)
0.0055-0.0060
(25° C)
0.071 (25° C)
1 x lO"1* (27° C)
0.09 (25° C)
300 (20° C)
Melting
Point
173-175° C
35-36° C
175-177° C
48.5-49.0° C
135-138° C
85-87° C

*not determined

-------
2)   Methyl Parathion  (Insecticide):   (0-0-dimethyl-0-p-nitrophenyl phos-
     phorothioate).Technical grade is a liquid.  Low solubility in water
     (Table 2), but soluble in most organic solvents.  Highly toxic and
     degrades readily  in a soil environment producing some metabolites that
     are equally toxic.  Acute oral LDso to rats is 9-25 mg/kg.
3)   Terbacil (Herbicide):  (3-tert-butyl-5-chloro-6-methyl uracil).
     soluble in water  (Table 2) and soluble in most organic solvents.
     mobile in soils due to relatively low adsorption.  Acute oral LD50 for
     rats between 5,000-7,000 mg/kg.
4)   Trifluralin (Herbicide):  (a,a,a-tri£luoro-2,6-dinitro-N, N-dipropyl-
     p-toluidine).Almost insoluble in water (Table 2) but very soluble
     in most organic solvents.  Volatile unless incorporated into the soil
     immediately following application.  Microbial and photo-degradation
     play a significant role in dissipation from soil.  Adsorption
     on organic matter and clay colloids retards leaching from soils.
     Acute oral LD^o for rats greater than 10,000 mg/kg.
5)   2,4-D (Herbicide):  (2,4-dichlorophenoxyacetic acid).  Somewhat soluble
     in water (Table 2), and very soluble (30-60%) in acetone and alcohols.
     Undergoes microbial degradation in soils, but losses due to photodecom-
     position are minimal.  Because of low adsorption in soils, it is
     readily leached.  "Acute oral LD50 for formulations are in the range of
     300-1000 mg/kg rats, guinea pigs, and rabbits.

     Formulated and technical grade materials fortified with 14C materials
were used to study mobility, adsorption-desorption, microbial degradation
and accumulation of metabolites in soils at various pesticide concentrations.
Stock solutions of each pesticide were prepared in 0.01N CaCl2 using the
commercial stock solution     made up to the aqueous solubility limit of
the pesticide.  Solutions of lower concentrations were prepared by succes-
sive dilutions of the  original stock solution.  A mixture of antibiotics
consisting of Penicillin G at 1 iug/ml and Polymixin B sulfate at 5 yg/ml
(Sigma Chemical Co., St. Louis, MO) was added to all pesticide solutions to
prevent microbial degradation during storage and use.

     The commercial formulation of 2,4-D was Ded-Weed 40 (Thomson-Hayward
Chemical Co., Kansas City, MO), a dimethylamine salt of 2,4-D (41% acid
equivalent).  The stock solution of commercially formulated atrazine was
prepared using AATREX  SOW (80% wettable powder; Ciba-Geigy Corp., Greens-
boro, NC).  A concentrated xylene solution of methyl parathion (80% solu-
tion; Monsanto Co., Agricultural Division, St. Louis, MO) was diluted in
0.01N CaCl2 to give the desired stock concentration.  The commercial form
of trifluralin was Treflan (Blanco Products Co., a division of Eli Lilly
and Co., Indianapolis, IN; 44.5% trifluralin and 55.4% inert ingredients).
The technical grade form of each pesticide was at least 97% pure.  All
pesticide solutions were spiked with the appropriate uniformly ring-labeled
™C compound (except for trifluralin) to give specific activities in the
range of 2-5 nCi/ml.   Trifluralin was 14C-labeled at the -CF3 position.

ADSORPTION ISOTHERMS

     Equilibrium adsorption isotherms for all soil-pesticide combinations
were 'measured using the batch procedure.  Equilibrium was achieved by

-------
shaking duplicate samples of 5 or 10 g of soil with 10 ml of pesticide
solution in Pyrex serew-cap glass test tubes for 48 hrs.  Preliminary
experiments had indicated that there was no measurable increase in pesti-
cide adsorption beyond this time.  Following equilibration, the test tubes
were centrifuged at 800 g for 10 minutes and the ^C-activity in 1-ml
aliquots of the clear supernatant solution was assayed by liquid scintilla-
tion counting.  Decreases in pesticide solution concentration were attri-
buted to adsorption by the soil.  All adsorption experiments were performed
at a constant temperture (23 ± 1° C).

PESTICIDE DISPLACEMENT THROUGH SOILS

     Pesticide movement through water-saturated columns of Webster, Cecil,
and Eustis soils was studied using the miscible displacement technique
described by Davidson et al. (1968).  Air-dry soil was packed in small
increments into glass cylinders (15 cm long; 45 on2 cross-sectional area).
Medium porosity fritted glass plates served to retain the soil in the
column. The soil was initially saturated with 0.01 N CaCl2 solution.  A
known volume of pesticide solution at a desired concentration was intro-
duced into the soil at a constant flux using a constant-volume peristaltic
pump.  After a specific volume of pesticide solution had been applied, the
pesticide solution was subsequently displaced through the soil column with
0.01 N CaCl2 at the same flux.  Effluent solutions were collected in 5 or
10 ml aliquots using an automatic fraction collector.  A pulse of 3H20
 (specific activity = 5 nCi/ml) was also displaced through each soil column
to characterize the transport of non-adsorbed solutes.  The activity of 11*C
and  3H in effluent fractions was assayed by liquid scintillation.  The
counting efficiencies exceeded 90% for 1!*C and 50% for 3H in all cases.

     The column experiments consisted of displacing 2,4-D amine solutions
at two concentrations (50 and 5,000 vg/ml) through columns of Cecil, Eustis
and Webster soils, and 5 and 50 pg/ml solutions of atrazine through a
Eustis soil.  All displacements were performed at a Darcy flux of approxi-
mately 0.22 cm/hr to ensure near-equilibrium conditions for pesticide
adsorption during flow.  The total volume of water held in the soil column
was gravimetrically determined at the end of each displacement by extruding
the soil from the glass cylinders and oven-drying.  The number of pore
volumes (V/V ) of solution displaced through the column was calculated by
dividing the cumulative outflow volume (V) by'total water volume (V ) in
the soil column.  Effluent pesticide concentrations are expressed a§ rela-
tive concentrations (C/C ), where C and C  are, respectively, effluent and
input concentrations.  Plots of C/C  versus V/V  are referred to as break-
through curves (or ETC).           °           °

     Air-dry soil -was packed into 3.2-cm diameter lucite cylinders composed
of 1-cm sections supported by a V-shaped container that permitted observa-
tion of the wetting front position with time.  Technical or analytical
grade pesticide was dissolved in benzene and was spiked with lkC-labeled
compound.   The benzene solution was mixed with air-dry soil (to give 200
or 2,000 yg pf pesticide/g of soil and 10 nCi/g soil) and the benzene
evaporated.   In order to simulate a waste disposal site, the pesticide-
spiked soil was packed into the top 1.5 cm of the soil column.  Infiltra-


                                     10

-------
tion of water into horizontal columns of soil was controlled by maintaining
the soil surface at a negative pressure (-4 on of water) using a fritted-
glass plate and a constant-head burette.  The fritted-glass plate apparatus
was filled with 0.01 N CaCl2 and the desired negative pressure applied
before it was placed in contact with the soil.  Measurement of time in all
experiments commenced the instant contact was established between the
fritted-glass plate and soil surface.  Water entering the soil was measured
volumetrically using a constant-head burette connected to the fritted-glass
plate apparatus.  Measurements of distance to the wetting front (zero at
the contact plane between soil and plate) were visually observed.  When
flow had proceeded for the desired time (i.e., until the wetting front had
advanced to about 30 cm), the water supply was discontinued and the soil
column was immediately cut into 1-cm segments.  About one-half of the soil
contained in each 1-cm segment was oven-dried at 105° C for 24 hours to
determine the gravimetric soil-water content.  The remaining one-half of
the soil from each 1-cm-segment was dried in a vacuum desiccator over P20s
or H^Oij for a 24-48 hour period.  About 0.5-0.7 g of the desiccator-dried
soil was then combusted in a Packard Model 306B sample oxidizer; the ll*C02
evolved by combustion was trapped in a premixed organic amine-fluor cocktail
and assayed by liquid scintillation.  The pesticide concentrations were
calculated using the specific activity  (dpm/yg pesticide) of the pesticide-
spiked soil sample.  The pesticide concentrations determined in the above
manner represent the sum of adsorbed and solution-phase concentrations and
were expressed as yg pesticide/g oven-dry soil.                   '

PESTICIDE DEGRADATION AND SOIL RESPIRATION

     Each soil was initially wet to a soil-water content corresponding to
30% of the 0.33 bar soil-water tension and incubated for one week at 25° C.
Following incubation, each soil was then mixed thoroughly with a specific
quantity of pesticide fortified with lkC-pesticide at 1 yCi/100 g of soil
and sufficient water was added to bring the soil up to 0-33 bar soil-water
tension.  For pesticide degradation and soil respiration experiments, 100 g
(oven-dry basis) of each pesticide-treated soil was placed in a 250 ml
Erlentneyer flask.  Special care was taken when mixing the pesticide with
the soil to ensure a uniform distribution of the pesticide within the soil.
Technical grade and formulated materials of each pesticide were used. The
flasks were then connected to a plexiglass manifold and C02-free water-
saturated air passed through the manifold into the falsks at a flow rate of
10 ml/min per flask.  For pesticides with high vapor pressures, the air
leaving the flask was passed through 40 ml of ethylene glycol to absorb the
volatilized pesticide.  The air leaving each flask was also bubbled through
a KOH solution (0.1-0.5N) to absorb the evolved C02.  At frequent intervals,
the KOH solutions were replaced with fresh KOH solutions and the total C02
concentrations in the KOH solutions were determined by titration.  lltC02
activity in the KOH solution was determined by liquid scintillation count-
ing.

SOIL MICROBIAL POPULATIONS

     For the microbial enumeration experiments, the experimental set-up
was essentially the same as that for the degradation experiment, except


                                    11

-------
 250  g of each pesticide-treated soil  (10 to  20,000 yg/g) was placed in a
 500  ml Erleraneyer  flask and no ltfC-labeled pesticide was used.  Ten gram
 soil samples were  withdrawn weekly from each fla'sk.  Bacterial, fungal and
 actinomycete populations  in the samples were determined using  a dilution
 plate count method.  The  number of microorganisms were expressed  as colony
 forming units  (cfu) per gram of oven-dry soil.  Bacterial populations  were
 determined using a TGY medium consisting of  5 g tryptone, 5 g  of  glucose,  4
 g of yeast extract, and 18 g of agar  in 1,000 ml of distilled  water. The pH
 of the medium after autoclaving was 7.0.  Bacterial colonies in the plates
 were counted after 48 to  52 hours at  28° C.   Fungi were enumerated in
 Martin's  (Martin,  1950) Rose Bengal-streptomycin agar (RBS agar).   The pH
 of the fungal medium after autoclaving was 6.0.  Fungal colonies were
 determined after 60 to 72 hours at 28  C.  Actinomycetes were  plated on
 starch-casein agar (SC agar) supplemented with antibiotics cycloheximide 50
 yg/ml and  nystatin 50 yg/ml (Williams and Davis, 1965).  The pH of the
 actinomycete medium after autoclaving was 7.2.  The actinomycete  colonies
 were counted after 12 to  14 days at 25° C.
         TABLE  3.   SOLVENTS USED FOR PESTICIDE EXTRACTION FROM SOIL
               Pesticide	Solvent	

           2,4-D                              Ether Ethyl

           Trifluralin                         1) Benzene/Ethyl Acetate
                                                 (3:1)
                                              2) MeOH

           Atrazine                            MeOH (soxhlet)

           Methyl Parathion                    Hexane: Ace tone (80:20)
METABOLITE IDENTIFICATION

     Soil treatments and incubation were essentially the same as for the
pesticide degradation experiments, except 150 g of soil were used and
2 yCi  lkC-pesticide per 100 g soil were added.  Ten gram soil samples were
withdrawn biweekly.  The soil samples were extracted three times with the
appropriate solvent (Table 3).  The 20,000 yg/g samples were extracted four
times because of their high pesticide concentrations.  The extracted soil
was then air dried and stored in a cold chamber prior to analysis for
"bound" ltfC.  The extracts were concentrated to 10 ml on a rotary evaporator,
or in a Danish Kuderna evaporator, then further concentrated using a gentle
stream of N2 and an aliquot was assayed for 1IfC.  The extracts were further
concentrated with N2 to one ml and an aliquot equivalent to 15,000 dpm
of each sample was streaked on a thin layer plate (see Table 4).  The TLC
plates were developed and placed on Kodak on-screen x-ray film  (NS-57)


                                    12

-------
for one month.  Radioactive streaks on the TLC plates were scraped and  the
radioactivity eluted with two ml of solvent  (Table 3) .  The percentage  of
radioactivity in each separate radioactive component on the TLC plates  was
determined by liquid scintillation counting.
     The unextractable "bound" portion of the  14C- residue was determined by
oxidizing the extracted soil samples in a stream of 02 at 800° C according
to the modified method of Watts  (1971) .  The air dried extracted soil was
placed directly in the oven at 800° C and the  02 stream which passed through
the soil was bubbled directly into 15 ml of phenethylamine C02- trapping
cocktail solution.  Complete combustion took approximately five minutes.
Then N2 was purged through the system to eliminate 02 and the trapped lkC02
was assayed.

     A Varian Aerograph Series 2100 gas chromatograph (GC) with one flame
ionization and three electron capture detectors was used for the gas chroma-
tography work.  2,4-D analysis consisted of quanitfication of the buto-
xyethyl ester of 2,4-D which was prepared as follows:  1) Five ml of acetyl
chloride was added dropwise to cold butoxyethanol (final volume equal to
100 ml) ; 2) 0 . 1 ml of this solution was then added to a test tube containing
the extracted 2,4-D.  The tube was sealed and  held at 80° C for 30 minutes;
3) The tube was cooled and 2.0 ml of hexane was added; 4) The hexane solu-
tion was then extracted three times with 0.2 M K2HPOit and 5) A small amount
of anhydrous sodium sulfate was then added to  the hexane solution to dry
the sample.  The GC column was 1.8mx2mmi.d. glass packed with 31 QF-1
80/100 on Gas Chrom Q and operated at 200° C.  The injection port and
detector temperatures were 250° and 300° C, respectively.  The same column
was used for trifluralin but was operated at 140° C.  Trifluralin and its
metabolites were assayed without modification.

     Atrazine and its dealkylated metabolites  were assayed using a 51 CBWX
20 M 60/80 mesh Gas Chrom Q packed inal.8mx2mmi.d. glass column and
utilizing a flame ionization detector.  The hydroxy metabolites of atrazine
were methylated and then assayed on the CBWX column using a flame ioniza-
tion detector.  Injector, column and detector  temperatures were 250 , 200
and 300° C, respectively.

     Mass spectral analyses for trifluralin were performed using a Finnigan
Model 1015C chemical inozation quadruple mass  spectrometer equipped with a
Varian 1400 gas chromatograph as the inlet source without a separator. The
mass spectrometer (MS) was interfaced to a System Industries 150 computer
data acquisition system.  The 1.8mx2mmi.d. glass column was packed
with 31 QF-1 on 100/120 mesh Gas Chrom Q.  Methane was used as the carrier
gas at a flow rate of 18 ml/min and passed directly into the ion source
where the pressure was maintained at 1.0 torr.  The injector, column and
transfer line temperatures were 240°, 140° and 230  C, respectively.

     In addition to mass spectra, the computer data system provided recon-
structed gas chromatograms (RGC) and limited mass range searches (LMS) .
RGC's represent the normalized total ion current plotted versus the spec-
trum number.
                                     13

-------
                          TABLE  4.  THIN LAYER CHROMATOGRAPHIC SYSTEMS
   Pesticide
    TLC Solvent Systems
    TLC Plates*
  TCL Scraping
Eluting Solvent
2,4-D


Trifluralin


Atrazine


Methyl Parathion
n-Butanol:Acetic Acid:H20
(4:1:1.8)

Hexane:MeOH  (97:3)
     :Acetone:Acetic Acid
(9:1:1)

Hexane:Acetone (80:20)
Silica gel 60F-254
(20 x 20 x 0.2 mm)

Silica gel 60F-254
(20 x 20 x 0.2 mm)

Aluminum Oxide
(20 x 20 x 0.2 mm)

Silica gel 60F-254
(20 x 20 x 0.2 mm)
Ether
Ethyl Acetate
CHCl3:Acetic Acid
(9:1)
*A11 were Brinkman

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                                  SECTION  5

                           RESULTS AND DISCUSSION
ADSORPTION AND MOBILITY

Adsorption Experiments

     Equilibrium adsorption  isotherms were determined for each soil-pesti-
cide combination by measuring pesticide adsorption at five or more concen-
trations ranging from zero to the pesticide's aqueous solubility.  All
adsorption isotherms considered in this study, with the exception of 2,4j%D
and the Glendale soil, were  described by the Freundlich equation (S = KCj,
where K and N are constants, and S and C are adsorbed (yM/kg soil) and
solution (yM/£) phase pesticide concentrations.  The values of the Freund-
lich adsorption constants, K and N, for each soil-pesticide combination
studied were obtained using  a least-square fit procedure to the adsorption
data.  These values are presented in Table 5.

     Equilibrium adsorption  isotherms should be independent of the soil to
solution ratio employed in the batch adsorption procedure.  This has not
always been true for some published data (Grover and Hance, 1970; Hance
1977).  The results presented in Figure 1 illustrate that no difference was
observed between the 2,4-D amine and Webster soil adsorption isotherms
conducted at soil:solution ratios ranging from 1:2 to 1:10.  The indepen-
dence of the adsorption isotherm to soil solution rate used in the batch
adsorption procedure was shown to be consistent for the other soil-pesti-
cide systems considered in this study.  These results are in agreement with
those of Green and Obien (1969), Nearpass (1967), and Dao and Lavy (1978).
This concept is important when using the Freundlich equation and its
constants, K and N, to predict pesticide partitioning between soil and
solution phases in systems where soil:solution ratios are known but not
constant (e.g., soil profile, runoff water containing sediments, etc.).

     The type and quantity of specific electrolytes in the water frequently
influence the adsorption of pesticides.  The sensitivity of pesticide ad-
sorption characteristics to electrolytes depends upon the adsorption mechan-
ism between the pesticide and solid phase.  For example, 2,4-D amine adsorp-
tion increases in the presence of calcium (Figures 2 and 3) for both Cecil
and Webster soils.  This phenomenon, however, is not consistent for all
soil-pesticide systems (Abernathy and Davidson, 1971).  Based upon the
results presented in Figures 2 and 3 one needs to be aware of the type and
quantity of electrolytes present in the water and their influence on pesti-
cide adsorption.


                                     15

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 TABLE 5.   FREUNDLICH CONSTANTS CALCULATED FROM EQUILIBRIUM ADSORPTION
           ISOTHERMS FOR VARIOUS SOIL-PESTICIDE COMBINATIONS
PESTICIDE
Atrazine




Methyl
Parathion




Terbacil
*



Trifluralin




2,4-D Amine



, ... .- .-• .
SOIL
Webster
Cecil
Glendale
Eustis
Average + % CV*

Webster
Cecil
Glendale
Eustis
Average ± 1 CV
Webster
Cecil
Glendale
Eustis
Average ± \ CV
Webster
Cecil
Glendale
Eustis
Average ± % CV
Webster
Cecil
Glendale
Eustis
Average ± % CV
f
9.12
0.84 '
0.69
0.85
2.87 ± 145

18.67
4.81
6.05
3.30
8.21 ± 86
2.96
0.39
0.42
0.15
0.98 ± 135
2.49
0.43
1.31
0.23
1.11 ± 92
7.27
0.84
--
1.14
3.08 ± 118
KG*
6.03
0.89
0.62
0.62
2.04 ± 131

13.39
3.95
3.57
2.72
5.91 ± 85
2.46
0.38
0.38
0.12
0.83 ± 130
2.93
0.46
1.60
0.24
1.31 ± 94
4.62
0.65
--
0.76
2.01 ± 112
N
0.73
1.04
0.93
0.79
0.87 ± 16

0.75
0.85
0.61
0.86
0.77 ± 15
0.88
0.99
0.93
0.88
0.92 ± 6
1.15
1.05
1.18
1.06
1.11 ± 6
0.70
0.83
—
0.73
0.75 ± 9
KJ
155.8
98.9
124.0
110.7
122.3 ± 20

346.0
438.6
714.5
486.4
496.4 ± 32
63.6
42.2
76.0
21.4
50.8 ± 47
75.7
50.7
177.8
43.2
86.8 ± 72
119.4
72.2 .
—
135.7
109.1 ± 30
     Freundlich constant when solution and adsorbed phase concentrations are
j-    expressed as pM/1 and pM/kg of soil.
 KG  Freundlich constant for solution and adsorbed phase concnetrations are
     as pg/ml and pg/g of soil [Kf = K-CMW/l.OOO)1^],  when MW is the pesti-
     cide's molecular weight.
     Freundlich constant for solution and adsorbed phase expressed as pg/ml
     and pg/g of organic carbon.
     is the coefficient of variation, I CV = (standard  deviation/average x
     100.
*cv
                                     16

-------
            1000
         en
         o
         E
        to

        u    100
        z
        O
        U
Q
U
ffl
CC
O
to

<    10
                 _ Webster  Soil

                     2,4-D Amine
•5=10.55 c
        P.62
                  SOIL:SOLN.
                                             o  1
                                             .  1
                                             •»•  1
                      2

                      5

                      10
                10                  100                1OOO

                      SOLUTION    CONC,   C  (jjmoles/1)
Figure 1.  Adsorption isotherm for 2,4-D amine and Webster soil.  Data ob-
          tained using soil:solution ratios of 1:2,  1:5 and 1:10.
                                 17

-------
        104
O)


"5




i/)

u
z
o
u
         10
      Q
      LU
      m
      a:
      O
               WEBSTER   SOIL

               2,4-D Amine
                                               s = 5.5? c
                                                      .07
                                                   o  CaCI2
           10'
                                                H2O
        101 	i  i  i 11 nil    i  i  i i n nl   i   i  i 11 nil    i   i i 11 MI
                       SOLUTION
                               icr           10

                              CONC., C  (jjmol/l)
1CT
Figure 2.  Adsorption isotherm for 2,4-D amine and Webster soil.   2,4-D was
          applied to soil in distilled water or 0.01 N CaCl-.
                                     18

-------
      10
O)


o





to

u
z
O
U
    Q
    U
    m
    cr
    O
    CO
    Q
       10
       10'
               CECIL   SOIL

               2,4-D  Amine
                                                         0.78
         10
                                                    2       —
          I   I  i i i ml    i   i  i i i iifl     i  i  i i i ml   i   i i  i i
                  10            10            10

                    SOLUTION   CONG., C   (jjmol./!)
Ill
 10
Figure 3.  Adsorption isotherm for 2,4-D amine and Cecil soil.  2,4-D was
          applied to soil in distilled water or 0.01 N CaCl2.
                                   19

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     Adsorption isotherms  for  atrazine, methyl parathion,  terbacil,  tri-
 fluralin,  and 2,4-D  amine  and  four soils are presented  in  Figures  4-8.
 Because  of the large quantity  of pesticide adsorbed by  the Terra Ceia muck,
 it was not included  in these figures.  The Freundlich adsorption constants
 which describe the data  for each isotherm are shown adjacent  to  the  best-
 fit line (least-squares) through the data.  Note that for  all pesticide-
 soil systems  studied, with the exception of trifluralin, N in the  Freundlich
 equation is less than one.

     To  achieve the  high solution concentrations of trifluralin  shown in
 Figure  7,  it  was necessary to  use a 1% solution of a nonionic surfactant
 (Triton  X-100; polyoxyethylene ethers).  Decreasing solution  concentrations
 of the surfactant due to its adsorption on the soil (Valoras  et  al.,  1969),
 could lead to a reduction  in pesticide solubility and promote precipitation
 of the trifluralin,  especially at the higher concentrations.  In this
 study, the amount of pesticide adsorbed by the soil was measured from the
 change in  solution concentration.  Thus, any decrease in pesticide concentr-
 ation in the  solution phase due to precipitation would have been erroneously
 attributed to adsorption by the soil.  If precipitation had occurred primar-
 ily at the higher trifluralin  concentrations, a value of N greater than 1.0
 would be anticipated.  It  should also be recognized that the  presence of
 surfactants can drastically increase or decrease pesticide adsorption on
 soils  (Hugenberger,  et al., 1972).  The surfactant may compete with  the
 pesticide  for adsorption sites on the soil, or it could enhance  the  lipo-
 phillic  nature or the soil surface and thereby increase pesticide adsorption.

     The adsorption  data for 2,4-D amine and the Glendale  soil are not in-
 cluded in  Figure 8 or Table 5  because a calcium salt of 2,4-D was formed in
 this soil  and precipitated out of solution making it impossible  to measure
 2,4-D adsorption at  large  herbicide concentrations.  The Glendale soil con-
 tains a  large quantity of  calcium carbonate.  The quantity of 2,4-D  amine
 precipitated  increased as  the  herbicide concentration in solution prior to
 contact  with  the soil increased.  The precipitation of 2,4-D  as  a calcium
 salt in  the Glendale soil  was  confirmed through analytical procedures (mass
 spectroscopy).

     Two important conclusions can be made based on the data  presented in
 Table 5.   First, the-fact  that the Freundlich equation describes all pesti-
 cide adsorption isotherms  over a wide concentration range  suggests that
 adsorption sites were not  saturated at any concentration considered  in this
 study.   The amount of pesticide adsorbed by the soil continued to increase,
 at  a decreasing rate, with each increase in solution concentration.  This
 behavior may not, however, hold for other pesticide adsorbents (Weber and
 Usinowicz,  1973).  Second, contrary to a frequent assumption, pesticide
 adsorption isotherms are generally nonlinear, that is, N is greater  or less
 than one (Table 5).  Linear adsorption isotherms have been generally accept-
 ed  for low pesticide concentrations because it simplified  computer simula-
 tion modeling  (Davidson, et al, 1968; Davidson and Chang,  1972; Hugenberger,
 et  al., 1972; Kay and Elrick,  1967).

     Because soil organic  carbon content generally correlates well with
pesticide  adsorption, the use  of an adsorption paratition  coefficient based


                                     20

-------
             io
                h  ATRAZINE
O)
^

o
E
          u
          a
          u
          CO
          a:
          O
          CO
          a
  10'
            1O
    10'
                                                WEBSTER


                                                 CECIL
                                                I   i  i i  I
                                                           10
                     SOLUTION    CONC.   (jumol/l)
Figure 4.  Adsorption isotherms for atrazine and Webster,  Cecil, Glendale
          and Eustis soils.  Freundlich constants (K and N)  for each iso-
          therm, determined by least-squares fit to the data,  are also
          shown.
                                   21

-------
     O)
     _*:

     "o

     E
     D


     (J
     u

     Q
     UJ
     CD
     CC
     O
     to
     Q
- METHYI  PARATHION
                                  WEBSTER
                                         ECIL
                                                     EUSTIS
                                          GLENDALE
                        i  i  f  i i i I
                                        I	i
                                     i   i i  i i  i 1
          10°                    101
                SOLUTION    CONC. (pmol./l)
                                              10'
Figure 5.  Adsorption isotherms for methyl parathion and Webster, Cecil,

          Glendale and Eustis soils.  Freundlich constants  (K and N) for
          each isotherm, determined by least-squares fit  to the data, are
          also shown.
                                 22

-------
          O)
          .*

          o
          E
          O
          o
          Q
          LU
          CO
          o:
          O
            10T
            10J
                 TERBAC
              10'
                                            WEBSTER
                                              EUSTIS
                                                I   i  i  i I  i i
1O
                     SOLUTION     C O NC. (jjmol./!)
Figure 6.   Adsorption isotherms for terbacil and Webster, Cecil, Glendale
           and Eustis soils.   Freundlich constants  (K and N) for each iso-
           therm determined by least-squares fit to data, are also shown.
                                   23

-------
           O)
           _*

           "o
           E
           3

           U102

           o
           u

           Q
           LU
           CD

           go'
           (/)
           Q
            1OC
                u TRIFLURALIN
WEBSTER



GLENDALE
i   i i  i i i
               10°           1O1           1O2

                     SOLUTION   CONC.(jumol./l)
                                      10J
Figure 7.  Adsorption isotherms for trifluralin and Webster, Cecil, Glendale

          and Eustis soils.  Freundlich constants (K and N) for each iso-
          therm, determined by least squares fit to data, are also shown.
                                  24

-------
       10T
     E
     u
     z
     O
     u


     Q
     LU
     CD
     Q:
     O
     to
     Q
       10'
          h   2,4-D Amine
                                                       CECIL
         10
l   i i  I i IIII    i   i  i i 1111
i  i I i 11 ll    I  l  I I  i 111
         1O"            10J            10*            1O

       SOLUTION     CONC.    (jjmoL/l)
Figure 8.  Adsorption isotherms for 2,4-D amine and Webster and Cecil soils.
          Freundlich constants (K and N)  for each isotherm, determined by
          least-squares fit to data,  are also shown.
                                    25

-------
upon organic carbon content rather than total soil mass has been proposed
by Lambert  (1968) and Hamaker and Thompson  (1971).  Using this procedure,
the amount  of pesticide adsorbed was expressed as yg/g organic carbon and
the Freundlich constant ifKp.p) for each adsorption isotherm was computed.
These values are also presented in Table 5.  It is apparent that the  values
of IC™  for  a given pesticide are much less variable (smaller percent  CV)
among the four soils studied than are the K values uncorrected for organic
carbon.  These results are in general agreement with the observations of
Hamaker (1975) where the K^ values for a given pesticide were nearly inde-
pendent of  soil type.  It should be recognized, however, that other factors
such as soil pH, clay content, and cation exchange capacity may  also  play a
significant role in determining pesticide adsorption by soils (Bailey and
White,  1970).  On the basis of the IC^, values listed in Table 5, the  extent
of pesticide adsorption in soils was in the order of terbacil <  trifluralin
< 2,4-D amine < atrazine < methyl parathion.

COLUMN  DISPLACEMENT EXPERIMENTS

     The partial differential equation generally assumed to describe  the
movement of pesticides and other adsorbed solutes through soils  under
steady-state water flow conditions is (Van Genuchten, et al., 1974):

                         3C   n 82C     9C   £ 3S
                         3t     3X2"     3X " 6 8t                       [1]

where t is  time (days), D is dispersion  coefficient (cm2/day) , x is dis-
tance (on) , v is average pore-water velocity (cm/ day) , p is soil bulk
density (g/on3) , e is volumetric soil-water content (on3/cm3) , and C  and
S are solution and adsorbed pesticide phase (yg/ml and yg/g) , respectively.
When the adsorption isotherm obeys the Freundlich equation, the  convective-
dispersive  solute transport model (Equation 1) reduces to:
                                         -                            '[2]

where ,

                         R(C)  =  [1 + pKNC^'1]                         [3]

     The retardation term R(C) is a quantitative index of the pesticide's
mobility in that its value is equal to the ratio of the positions of the
adsorbed and nonadsorbed solute fronts in soil.  The value of the adsorp-
tion coefficient K in Equation [3] for nonadsorbed solutes (e.g., chloride
or 3H20) is equal to zero; hence, R(C) = 1.  For adsorbed solutes, R(C) is
greater than one because the value of K is larger than zero.  Thus, larger
values of R(C) indicate reduced pesticide mobility in soils.  It may be
noted from Equation [3] that for the case of nonlinear adsorption isotherms
(N < 1) , the value of the retardation term increases with decreasing solu-
tion concentration C, while for a linear isotherm (N = 1), R(C) is indepen-
dent of pesticide solution concentration.  Thus, the mobility of pesticides
and other adsorbed solutes through soils is directly influenced by the
shape of the equilibrium adsorption isotherms.

                                      26

-------
     Effluent breakthrough curves  (ETC) were measured for  2,4-D amine  at
two input concentrations  (C  = 50  and  5,000 pg/ml)  and tritated water
(3H20) using columns of Webster, Cecil and Eustis soils.   These ETC  are
shown in Figures 9, 10 and 11.  Tritiated water  represents a nonadsorbed
solute and serves as a reference for the  adsorbed solutes  (2,4-D amine in
this case).  A shift of the ETC for adsorbed solutes  to the right of 3H20
ETC is due to an adsorption-induced retardation.  The greater the right-
hand shift of the ETC, the greater the adsorption;  thus, a decrease  in
mobility. It is apparent  from the  data presented in Figures 9,  10 and  11
that the mobility of 2,4-D amine was significantly  increased as the  input
concentration (C ) increased from  50 to 5,000 yg/ml.   Note that for  the
5,000 yg/ml inpu£ concentration, 2,4-D amine was nearly as mobile as was
3H20. The effect of increased mobility at high concentration was more
pronounced in the Webster soil (Figure 9)  than in the Cecil (Figure  10) or
Eustis soil (Figure 11).  These column results are  consistent with Equation
[3] and the measured nonlinear adsorption isotherms (Table 5) for 2,4-D
amine.

     Breakthrough curves  for the displacement of atrazine  through the
Eustis soil at two herbicide input concentrations (C   = 5  and 50 yg/ml) and
3H20 are presented in Figure 12.   The  trend of increased mobility at higher
atrazine solution concentrations again is evident.  However, the differ-
ences in pesticide mobility between the two concentrations were not  as ,
large for atrazine (Figure 12) as  they were for  2,4-D amine (Figures 9-11).
Deviations of the adsorption isotherms from linearity,  i.e., constant
retardation term, increase exponentially  as the  concentration differences
become larger and/or as N approaches zero (Davidson,  et al., 1976; Hamaker
and Thompson, 1972).  In  this study, atrazine concentrations varied  only by
ten-fold while the 2,4-D  amine concentrations varied  by 100-fold.  Further-
more, the isotherm for 2,4-D amine adsorption in Webster soil was more
nonlinear than that for the Eustis soil-atrazine system (Table  5).

     The position of the  ETC for an adsorbed solute is  governed by the
nature of the equilibrium adsorption isotherm (Equation 3), whereas  the
shape of the ETC  (i.e., symmetry or lack  of it)  is  defined by nonlinearity
of the adsorption isotherm and the kinetics of adsorption-desorption pro-
cesses.  Symmetrical ETC  are obtained  when adsorption is an instantaneous
process and the adsorption isotherm is linear.   For nonequilibrium adsorp-
tion conditions during flow, asymmetrical ETC are generally obtained (Van
Genuchten, et al., 1974;  Rao, et al.,  1979).  All of  the pesticide ETC
measured in this study (Figures 9-12)  were asymmetrical in shape with
extensive "tailing" observed as C/C  approached  1.0 or zero.  Tailing was
absent in 3H20 breakthrough curves.0 The  extent  of  the asymmetrical  shape
of each pesticide ETC exceeded that which could  be  attributed to the non-
linear nature of the adsorption isotherms.  Hence,  much of the  asymmetry
measured for the pesticide ETC may be  attributed to nonequilibrium condi-
tions which.exist in the  soil columns  during flow.  Rao et al.  (1979)
presented an evaluation of two conceptual models where nonequilibrium
during flow was attributed to either kinetics-controlled or diffusion-
controled adsorption-desorption processes.
                                     27

-------
    i

  D

  ~ 0.8
  u
  z
  o
  <-> 0.6


  LU
  > 0.4
  £0.2
   o.o
    8J*-C0=5000jjg/ml
     o


     o
HO
      o

       o ,
                         • "
                                  '°Oc|oO o oOol no
                                WEBSTER  SOIL

                                 2,4-D   Amine
      A         6         8


      PORE    VOLUMES   (V/V)
                                                       10
12
14
Figure 9.  Effluent breakthrough curves for  2,4-D amine (Co = 50 and 5,000

           yg/ml) and for tritiated water displacement through Webster soil
           column.
                                     28

-------
  1.0
u
NmiX

U
z
O
U
  08
0.6
UJ 0.4



5 0.2
UJ
a:
  0.0
                *+ Ow00oo0ood0oooo0oo0<*>o
                B<*               c
                    •~c»
            o    •
                                 X
                          0 = 5000jug/ml
                                     +
  I         I
CECIL   SOIL
2,4-D  Amine
                                                          I
                               3456
                               PORE    VOLUMES  (V/VJ
                   8
Figure 10.  Effluent breakthrough curves for  2,4-D amine  (Co =  50 and  5,000
            yg/ml) and for tritiated water displacement through Cecil  soil
            column.
                                      29

-------
  y°
  u
    1.0
00.8
O
U0.6


>°'4
I-
< 0.2
LiJ
ct:
       —    o
                                           EUSTIS   SOIL
                                           2,4-D   Amine
                    000°
                             oooooo

              *»
                                                      50 jug/ml
                          C0= 5000jug/ml-v»o
                                          o
                                           o
                              I
                                   I
f *
                  2468
                         PORE    VOLUMES  (V/VJ
                                                               10
Figure 11.  Effluent breakthrough curves for 2,4-D amine (Co = 50 and 5,000
           vg/ml) and for tritiated water displacement through Eustis soil
           column.
                                  30

-------
  6          8
VOLUMES
                                                       (V/V0
Figure 12.  Effluent breakthrough curves for atrazine (C0 = 5 and 50 ug/ml)
            and for tritiated water displacement through Eustis soil column.
                                     31

-------
     The  illustrated increase in pesticide mobility at high concentrations
 limits the usefulness of the present low concentration data base for  devel-
 oping safe management practices for pesticide disposal in the soil. However,
 underestimation of pesticide movement by assuming linear adsorption isotherms
 may not be severe for pesticides with low aqueous solubilities. Ou et al.
 (1978a, 1978b) showed that for high loading rates, up to 20,000 yg 2,4-D/g
 soil, there was a significant decrease in the pesticide degradation rate
 with a concomitant depression of total microbial activity in the soil.
 Thus, due to rapid leaching and minimal microbial decomposition of pesticides
 at  high concentrations, the potential for groundwater contamination with
 pesticides is increased.

 SIMULATION OF PESTICIDE MOBILITY IN SOILS

     Several conceptual models have been proposed and evaluated for describ-
 ing the solute adsorption-desorption term (9S/3t) in Equation [1]. Because
 many of these models are based on the assumption of instantaneous adsorption
 and linear isotherms, they fail to describe experimental data (Davidson, et
 al., 1976).  Models based on first-order or other reversible kinetic  adsorp-
 tion-desorption processes were found to simulate experimental data reason-
 ably well at low pore-water velocities but failed at high pore-water  veloci-
 ties (Davidson and McDougal, 1973).

     Recent attempts to model the asymmetry or "tailing" in experimental
 breakthrough curves thought to be associated with nonequilibrium adsorption
 may be classified into two groups. In the first group, physical processes
 are assumed responsible for the observed tailing. In these models, the
 soil-water regime was divided into mobile and immobile regions. Although
 the solute adsorption-desorption was assumed to be instantaneous, the rate
 at  which  adsorbent molecules approached a fraction of the adsorption  sites
 was governed by diffusion through the stagnant soil-water region (Skopp and
 Warrick,  1974; Van Genuchten and Wierenga, 1976).  The assumption that a
 wide-range in pore-water velocities would result in the observed tailing
 has been  evaluated by Rao et al. (1976), using a capillary bundle model.
 In  a variation of the latter approach, Skopp et al. (1977) considered
 convective-dispersive solute transport in a system consisting of two  soil-
 water phases where the mass transfer of the solute between these phases
 obeyed a  first-order kinetic process.

     In the second group of conceptual models, the observed excessive
 asymmetry in the solute breakthrough curves was attributed to chemical
processes.  Adsorption-desorption isotherms for these models were assumed
nonsingular (Van Genuchten, et al., 1974; Hornsby and Davidson, 1973).
Because these models assume instantaneous equilibrium between the adsorbed
and solution phases, they failed to describe experimental data for high
pore-water velocities.  Furthermore, the physical and/or chemical justifi-
cation for the nonsingularity in the adsorption-desorption isotherms  has
been questioned.   More recently, Selim et al. (1976) and Cameron and
Klute (1977)  have proposed to two-site adsorption-desorption model for
describing asymmetrical breakthrough data.  Because nonsingular adsorption
isotherms observed earlier could be simulated using this model (Davidson,
et al., 1976; Selim, et al., 1976) and because justifications for such a

                                     32

-------
conceptualization may be found in the  chromatography  literature  (Giddings,
1965) , a thorough evaluation of the two-site  adsorption-desorption model
appears warranted.

     Agreement between model simulations  and  experimental data is generally
used as a criterion for verification of conceptual models.  Davidson et al.
(1976) discussed the limitations of such  an approach  when model parameters
were estimated by "best-fit" to experimental  data and not by independent
measurements.  However, due to the present inadequacy of experimental tech-
niques, independent determination of model parameters may not always be
feasible.  However, for curve- fitting  procedures to be valid for model
verification purposes, the same set of parameters estimated from a given
experiment should be used to predict experimental results obtained under
different conditions (e.g., column length and input concentrations, etc.).

     In the model proposed by Selim et al. (1976) to  describe the adsorption
term 3S/3t in Equation  [1J , two groups of adsorption  sites were assumed to
be responsible for solute adsorption by soils; one group of sites achieved
instantaneous equilibrium (type-1) while  the  other group was time- dependent
(type -2).  Using this approach, the time  rate of change in the adsorbed
phase concentration (3S/3t in Equation 1) may be expressed as,
                p.  3S =  fpKiNCj   3C  +

                e   9t        e        at             e                    [4]

where KI and N are  constants associated with  instantaneous adsorption  on
type-1 sites, lq  and k2  are, respectively,  forward and backward rate coeffi
cients (day *) for  kinetic  adsorption on the  type- 2 sites, S2 is adsorbed
phase concentrationon the kinetic sites, and  other terms are as defined
previously.  Note that at equilibrium,  total  adsorption is the sum of  the
two types of sites  and described by  the Freundlich equation:
               S = Si + S2 = K^ + K2CN  = KC^                         [5]

where K =  (K]. + K2) , Klf K2, and N are Freundlich constants, Sj and S2
(yg/g) are adsorbed phase concentrations  on  type-1 and type -2 sites, res-
pectively, and.K and S are defined on the basis of total solute adsorbed.
Note that the exponent N in Equation  [5]  was assumed identical for both
sites.  Assuming that type-1 sites represent some fraction F of the total
available adsorption sites, the following relationships may be stated:

                          KI = FK                                     [6a]

                          K2 = (l-F)K                                 [6b]

     By assuming a linear equilibrium adsorption isotherm  (i.e., N = 1 in
Equations  [4] and [5]), an analytical solution to the two-site adsorption
model (i.e., Equation [4] coupled with Equation  [1]), has been obtained by
Cameron and Klute (1977) .  Numerical solutions for nonlinear adsorption
isotherms as well as the more general case when adsorption on both sites is
kinetic-controlled were presented by Selim et al.  (1976).


                                     33

-------
      In  the  conceptual model  (Equation  [1]),  the values  of the experimental
 variables K, N,  6,  p, and v are known.  The dispersion coefficient,  D,  for
 each soil was  estimated  from  tritiated water  breakthrough  curves  using  the
 method proposed  by  Rose  and Passioura  (1972).  Experimental methods  are
 unavailable  to independently  measure the values of F,  KI and K2.   It may be
 noted from equations  [4],  [5]  and  [6] that,

                    K2 =  (1  -  F)K =  (6k1/pk2)                           [7]

 and
                    k2 =  6k!/(1 - F)PK                                  [8]

 Since the value  of  k2 can be  calculated given K, F, and kl5 the problem now
 reduces  to that  of  estimation of the two unknown parameters F  and ki.

      The model parameters were estimated using a nonlinear least-squares
 (NLLS) optimization procedure (Meeter and Wolfe, 1968) to  fit  the model
 prediction to  measured ETC  at low input concentrations.  This  iterative
 technique is based  on minimizing the differences between simulated and
 measured ETC data by successive refinement of the  initial  parameter  values;
 the estimates  at each iteration are obtained  by a  combination  of  Gaussian
 method and steepest descent method described  by Marquardt  (1963).  These
 estimates of model  parameters were then used  to verify the conceptual
 models by comparing the  predicted and the measured ETC obtained at a higher
 input concentration.  Conversely, the model parameters obtained by fitting
 to high  concentration ETC data were also used to predict low concentration
 ETC data.  Model verification procedures similar to this approach have  been
 employed by  Gaudet  et al.  (1977), O'Connor et al.  (1976),  and  Van Genuchten
 and Wierenga (1977).

      A finite-difference scheme was used to solve  the  model (Equations  [1]
 and [4]) subject to the  following initial and boundary conditions:

                  C = 0, S = 0,  0£X£L,  t =  0                   [9a]

                                 vCQ,  x = 0,  t £ t:

                                                                      [9b]

                                 0, x = 0, t  > ti


                  ax'"'        X = L>         t>0                 [9C]

These  conditions  are applicable for a soil column  of L length  (cm),  initially
void of pesticide, to which a pesticide solution of C   (yg/ml)  concen-
tration is applied at an average pore-water velocity of v  (on/day) for  a
period of ti days, and then leached through the soil with  a pesticide-free
solution.  All computations were performed on an AMDAHL  470 V/6-11 digital
computer (soft-ware compatible with IBM 370/165 systems) with  the aid of
computer programs written in  FORTRAN IV language.
                                     34

-------
     Measured and simulated ETC  for  2,4-D  amine  displacement  through a
water-saturated Cecil soil column  are presented  in  Figure  13.  Pesticide
mobility was greater for high  input  concentrations  as  indicated by the
left-hand shift of the ETC for C  =  5,000  yg/ml  compared to that for C  =
50 yg/ml. This increased pesticide mobility  at higher  concentrations i§ due
to the nonlinearity of the adsorption isotherm (Rao et al.,  1979).  The
simulated curves, shown as solid lines  in  Figure 13, were  calculated using
the two-site model, where the  parameters F and kx were estimated by curve-
fitting to the C  = 50 yg/ml ETC data.  Independently  measured values of K
and N (Table 5) along with the estimated values  of  F,  k1}  and k2 were used
to predict the ETC for C  = 5,000  yg/ml.   The agreement between the simula-
tions (solid lines in Figure 13) and the measured ETC  is only fair.  There-
fore, the model parameters K,  N, F,  ki, and  k2 were re-estimated using a 4-
parameter fit procedure and the  C  = 50 yg/ml ETC data.  Improved prediction
(dashed lines in Figure 13) of the1 measured  ETC  using  the  four parameter
fit procedure is evident.

     The values of the model parameters K, N, F, ki, and k2 estimated from
2,4-D amine data for C  = 50 yg/ml by varying two parameters  (F and kj or
four parameters (K, N,°F and ki) in  the NLLS procedure are presented in
Table 6.  More than a two-fold increase in K value  and a small decrease in
N over that obtained from an equilibrium adsorption isotherm was necessary
to describe the ETC data for both  input concentrations.  Furthermore, the
two estimates of the kj and k2 values were also  different.  Note that about
60 to 70% of the total adsorption  sites were required  to be kinetic (i.e.,
type-2 sites) in order to describe the  extensive tailing in the measured
ETC;

     unlike the ETC for the pesticides, those for 3H20 displacement through
the three soils were symmetrical and sigmoidal in shape (see  Figures 9, 10,
and 11).  Excellent descriptions of  the ETC  were obtained  using the convec-
tive-dispersive transport model  (Equation  1) where  all the soil-water was
assumed to be mobile.  A conceptual  model  where  the soil and  soil-water are
partitioned into mobile and immobile phases  failed  to  describe the ETC for
^H20 for the. three soils studied (Rao   et  al., 1979).  These results suggest
that while Equation [1] correctly  represented nonadsorbed  solute transport
processes, the nonequilibrium  adsorption-desorption phenomenon were not
adequately described by Equation [4].  Additional studies  need to be carried
out to identify the causes of  nonequilibrium conditions for pesticide
adsorption-desorption during transport  in  soils.

INFILTRATION EXPERIMENTS

     Pesticide transport in soils  during transient, unsaturated, and one-
dimensional water flow was investigated using hand  packed  horizontal soil
columns.  In order to simulate a waste  disposal  site,  the  top 1.5 cm of
each soil column was packed with pesticide-treated  soil (2,000 yg pesti-
cide/g soil), and 0.01 N CaCl2 was infiltrated into the soil  at a constant
negative head (-4 cm of water).  During infiltration,  the  rate of wetting
front advance was recorded by  visual observations.  The soil  column was
cut into 1-cm segments at the  end  of the infiltration.  In each soil seg-
ment, the total amount of pesticide  (sum of  solution,  adsorbed, and solid

                                      35

-------
TABLE 6.  COMPARISON OF MODEL PARAMETER VALUES EXTIMATED FROM THE 2,4-D
          AMINE BREAKTHROUGH DATA (C  = 50 yg/ml) FOR CECIL SOIL BY
          VARYING EITHER TWO OR FOUR°PARAMETERS IN THE NONLINEAR LEAST-
          SQUARES CURVE-FITTING PROCEDURE
                                      CECIL -2,4-D AMINE
PARAMETER
N
K
ki
F
kiCday"1)
k2(day-i)
klA2
2 -parameter Fit
0.83*
0.664*
0.324
0.488
0.271
0.222
. 1.220
4-parameter Fit
0.73
1.604
0.425
0.265
0.515
0.122
4.221

*These values of the Freundlich constants were obtained independently
 from the equilibrium adsorption isotherm.
                                     36

-------
   1.0
u
z
o
u
0.8
  0.6
U
K
        CECIL  SOIL
        2,4-D Amine
                                        *S
                                                TWO-SITE  MODEL

                                                K=0.664, N = 0.83; F= 0.488
                                                  =0.271 day4. k2= 0.222 day"1
{:
                                                <=1.604 , N = 0.73,- F=0.265
                                                 f 0.515 day1.  k2= 0.122 day"1
                                                           ooo 50  jug/ml
                                                           •••• 5000
                                          VOLUMES
Figure 13.  Measured and simulated breakthrough curves for 2,4-D amine dis-
            placement through Cecil soil column.  Parameter values used to
            calculate the solid lines were obtained from a 2-parameter fit,
            and those for dashed lines were estimated from a 4-parameter
            fit to C0 = 50 pg/ml data.
                                    37

-------
phases) was determined by combustion, while the soil-water content was
measured by oven-dry ing.

     The depth (X ) to which the pesticide front moved in a soil due to
water infiltration* was dependent upon the wetting front depth  (XJ, the
pesticide adsorption isotherm constants (K and N), and the aqueous solubi
lity (C ) of the pesticide.  The relationship between these variables may
be exprlssed as:
                          Xw       pl
                           w s 1 + —^_
                                     e                                 [10]
where, p is the soil bulk density (g/cm3) and e is the average soil-water
content  (cm3/cm3) in the wetted zone behind the wetting front.  It should
be noted that for the case of a linear adsorption isotherm (N = 1), Equa-
tion  [10] is exact and the retardation of the pesticide front due to adsorp-
tion  is independent of concentration, while for the nonlinear isotherm
case, Equation [10] is only an approximation.

     Assuming that Darcy's law is valid for unsaturated water flow in
soils, the rate of the advance of the wetting front is given by (Kirkham
and Powers, 1972):

                            Xw = mt1/2                                [11]

where, m is a constant and t is time (min).  In the present study, Equation
[11]  described (r2 >_ 0.95) the advance of the wetting front for all soil
columns considered.  The values of m and other pertinent data for the
infiltration experiments are summarized in Table 7.

     Measured pesticide concentration profiles and soil-water content
distributions at the end of infiltration in Eustis,'Cecil, and Webster soil
columns are shown in Figures 14 and 15.  Because the final wetting front
position in each soil column was different, for ease of comparison the
ordinate in Figures 14 and 15 is plotted as soil depth relative to the wet-
ting  front depth  (i.e., X/XJ.  Except for the 2,4-D-Eustis and the terbi-
cil-Eustis data, the relative mobilities of the pesticides are in general
agreement with those anticipated from the equilibrium adsorption isotherms
and pesticide aqueous solubilities.  The measured mobility of terbacil and
2,4-D in the Eustis soil (Figure 14) was nearly the same although the
adsorption coefficient for 2,4-D is greater than that for terbacil in
Eustis soil (see Tables .5 and 7).  The importance of aqueous solubility is
demonstrated by the atrazine data in the Eustis soil (Figure 14 and Table
7). Note that the .volume of water infiltrated into the soil column could
solubilize and transport only about 4% of the total pesticide present in
the top 1.5 cm segment; thus, most of the atrazine does not appear to have
moved.  The retardation factors, (X/X ), calculated by Equation  [10] are
generally larger than those measured in the infiltration experiments
(Table 7).  Similar results were reported by Wood and Davidson  (1975) for
transient-flow studies and by Davidson and Chang (1972) for saturated flow
experiments.  The kinetics of pesticide adsorption-desorption in soils are


                                      38

-------
                          TABLE 7.  PHYSICAL DATA FOR INFILTRATION EXPERIMENTS
vo

SOIL
Eustis
Eustis
Eustis
Cecil
Webster
PESTICIDE
Atrazine
2,4-D
Terbacil
Terbacil
Terbacil
CY
_ P .A.
S W
(pg/ml) (g/cm3) (cm)
33
650
710
710
710
1.67
1.69
1.64
1.51
1.40
21.3
30.0
29.5
28.4
24.1
m*
\/\
(on/min%) Measured
3.195
3.854
4.039
0.705
0.543
2.
1.
1.
1.
4.
50
05
11
49
16
Calculated*
3.
1.
1.
3.
4.
03
89
37
03
55
Volume
of water
applied
* (ml)
44
68
63
68
67
Pesticide
Recovery
(*)
>100
>100
91
93
99
v
     *see Equation [11]



     **see  Equation [10]

-------
           Pesticide   Concentration, jjg/g  soil
                250    0	250   0          250	
   Q2
   0.4
 JC
 +->
 Q.
  1.0
 
-------
                    TERBACIL    CONCENTRATION, jjg/g soil
                                                              500
                                                                       75O
            0.2
                   O.3    0.1      0.2

                 SOIL-WATER
    0.3      0     0.1

CONTENT ,  cm3/cm3
                                                           0.2
                                                                  O3
                                                                         0.4
Figure 15.  Soil-water content (solid lines)  and terbacil concentration
            distributions  in Eustis,  Cecil and Webster soils following
            infiltration of water to  approximately 30-cm.  Soils were
            initially air  dry and herbicide was in top 1.5-cm of soil
            (2,000 yg/g of soil).
                                     41

-------
not understood well enough at this time  (Rao et al., 1979) to describe
modeling the nonequilibrium conditions for pesticide adsorption-desorption
during transient soil-water flow.  Additional studies are needed in this
area.

MCROBIAL ACTIVITY AND DEGRADATION

     Total C02 evolution  (respiration) is generally a good indicator of
soil microbial activity.  This procedure was used by Stojanovic et al.
(1972) to estimate pesticide degradation rates in soil systems receiving
large pesticide concentrations.  A more direct procedure for determining
pesticide degradation, however, is to measure ^CC^ evolution from uniformly
ring-labeled  (llfCF3 position for trifluralin) pesticides.  In this study,
the accuracy of total C02 evolution as an estimate of pesticide degradation
was compared with that obtained using ll*C02 evolution.

ATRAZINE

     Soil respiration rates from Cecil soil treated with 10 ppm of technical
grade atrazine generally were not different from the soil without atrazine
during the entire incubation period of 82 days (Figure 16).  Soil respiration
rates for the 1,000 and 20,000 ppm treatments were generally smaller than
the untreated soil with the inhibition being the greatest for the 20,000
ppm treatment.  Total C02 evolution from the Cecil soil receiving 10, 1,000
and 20,000 ppm of formulated atrazine was enhanced, especially during the
first 16 days of incubation and the stimulation was greatest for the 1,000
and 20,000 ppm treatments.  Both technical grade and formulated atrazine at
concentrations of 10, 1,000 and 20,000 ppm enhanced the C02 evolution from
the Webster soil (Figure  17), with C02 evolution being greater for the
largest herbicide concentration.

     For incubation periods up to 70 days, 80-90% of the 1£tC-activity
extracted from the Webster soil was in the form of intact atrazine; the
remainder of the * ^-activity existed as metabolites.  More extensive
break-down occured in the soil treated with 10 ppm of technical grade
atrazine and then only after incubation exceeding 80 days.  A small portion
(less than 1%) of the applied 1£*C from all treatments was lost as lkCQ2
during the entire incubation period.

     The breakdown of atrazine in Cecil soil was more rapid and more exten-
sive than it was in the Webster soil.  However, the amount degradated de-
clined with increasing concentrations.  In the case of the 10 ppm treatment,
there was a substantial breakdown of atrazine.  After 63 days of incubation
of the technical material, 14% of the llfC remained as the parent compound;
for the case of formulated atrazine incubated for 75 days, 40% was intact
atrazine.  Cecil soil treated with 1,000 ppm of atrazine exhibited somewhat
less breakdown.  After 55 to 63 days of incubation with the technical grade
material, 70 to 82% of the ll*C extracted was parent atrazine; and after 75
days of incubation with the formulated herbicide, 50% of the herbicide
was unchanged.  In the Cecil soil fortified with 20,000 ppm, even less
breakdown occurred; after incubation for 75 days, 80% of the lltC remaining
was atrazine.  Similar to the Webster soil, only a small quantity of the


                                      42

-------
      _   A
o
O)
O
o
    6
E  4
o
 cvj
8a
o
               20
o—o      o Ppm
•—•     10 ppm
°—a  1,OOO ppm
   120,OOO ppm
                        40
       60
                      Days
                                                8
                                                0
                                                     1     I
                                                     B
20
                                                                                O ppm
                                                                               10 ppm
                                                                       a—°  1,OOO ppm
                                                                       •—• 2O.OOO ppm
40
                                                                  Days
60
Figure 16.  C02 evolution rate from the Cecil soil receiving various concentrations of atrazine.
           (A) Technical grade; (B) Formulated atrazine.

-------
    3
 m
 O)
 O
 O
 D)
o
 CM
O
u
                         o—o      O ppm
                         •—•     10 ppm
                         t>-a  1,000 ppm
                         •—• 2O.OOO ppm
    O
20
   40
Days
60
                                      B
                                              o—o     0 ppm
                                              •—•    1 0 ppm
                                              o—a 1.OOO ppm
                                              •—• 20,000 ppm
0
2O
   40
Days
60
Figure 17.  C0£ evolution rate from the Webster soil receiving various concentrations of atrazine.
          (A) Technical grade; (B) Formulated atrazine.

-------
total lkC-activity (less than 1%) was degraded to  14C02 in the Cecil soil.

     Technical grade and formulated atrazine at concentrations of 10, 1,000
and 20,000 ppm did not have a significant effect on bacterial populations
in the Cecil or Webster soils, except for the 20,000 ppm formulated atrazine
concentration in the Cecil soil  (Table 8).  The bacterial population in
this treatment was nearly four times higher than the untreated soil following
eleven weeks of incubation.

     The number of fungi in the  technical grade treated Cecil soil at 10,
1,000 and 20,000 ppm were not significantly different from the untreated
soil following eleven weeks of incubation (Table 9).  Fungal populations in
the technical grade treated Webster soil at the same application rates
were, however, significantly higher (p < 0.01).  Fungal populations in all
the formulation atrazine treated soils, but one, were not significantly
different.  Fungal populations in the Cecil soil treated with 20,000 ppm of
formulated atrazine were reduced significantly (p  < 0.01) during the first
seven weeks of incubation.

     Technical grade and formulated atrazine at concentrations of 10, 1,000
and 20,000 ppm showed no consistent effect on the  actinomycete populations
in the Cecil or Webster soils, except for the Cecil soil which received
20,000 ppm of formulated atrazine (Table 10).  In  this treatment, actino-
mycete populations were reduced  significantly (p < 0.01).

METHYL PARATHION

     Total C02 evolution from the Webster soil containing 24.5 and 10,015
ppm of technical grade and formulated methyl parathion were generally not
different from an untreated soil (Table 11), except during the first few
days of incubation.  C02 evolution was somewhat greater for the treatments
receiving a pesticide during the first few days of incubation than for the
untreated soils.  Unlike the Webster soil, total C02 evolution from the
Cecil soil containing 10,015 ppm of methyl parathion was reduced (Table
12). The reduction was somewhat  greater for the Webster soil receiving the
formulated material.  Apparently, the formulation  chemicals at this concen-
tration exhibited an inhibitory  effect on soil respiration in the Cecil
soil, but not for the Webster soil.  C02 evolution from the Cecil soil re-
ceiving 24.5 ppm of technical grade or formulated methyl parathion was not
different from the untreated soil.

     Both technical grade and formulated methyl parathion at 24.5 ppm were
degraded rapidly to C02, H20 and simple inorganic  ions in the Webster and
Cecil soils as indicated by 14C02 evolution from uniformly ring labeled
1HC-methyl parathion (Table 13).  More than half of the ltfC -methyl parathion
added was mineralized to ll*C02 in 10 days in the Webster and Cecil soils.
Degradation rates were greater in the Webster soil than the Cecil soil.
Degradation rates from the soil  receiving the technical grade material were
somewhat greater than those receiving the formulated material.  At the end
of 52 days of incubation, 75% to 85% of 1IfC-methyl parathion in the 24.5
ppm treatments was degraded to 14C02.  Lichtenstein, et al. (1977) reported
that up to 43% of the 14C-ring labeled methyl parathion applied to a soil


                                      45

-------
.£>.
ON
    TABLE 8.   EFFECT OF TECHNICAL GRADE AND FORMULATED ATRAZINE ON BACTERIAL POPULATIONS IN CECIL AND
               WEBSTER SOILS
     Concentrat ion
     of atrazine                                        cfu-g'1  soil  (x  10"6)
     (vg*g l)                                                Time (Weeks)
                                  0.1            1              23

0


10


Cecil

Webster
Cecil

Webster
Cecil
26.0a(26.4)b

47.6(53.8)
24.9(29.9)

58.4(56.2)
30.1(30.6)
16.2(20.5)

31.7(30.7)
21.9(23.3)

33.6(32.2)
18.0(20.5)
17.3(19.6)

28.3(22.4)
20.1(19.2)

31.7(28.9)
18.0(24.9)
14.4(13.0)

ND (28.6)
12.5(16.4)

ND (28.3)
10.3(14.8)
19.2 (ND)

20.5(24.7)
13.3 (ND)

22.1 (30.9)
10.9 (ND)
15.5(18.7)

18.6 (ND)
14.1(14.8)

24.3 (ND)
17.3(15.0)
        1,000
                    Webster   62.7(30.4)      33.8(38.2)    22.9(25.7)    ND  (20.3)    22.6(34.8)    17.7  (ND)

                    Cecil      32.4(69.1)d    19.6(111.0)d 22.1C(121.0)d14.4(70.9)d   16.2  (ND)     18.5(51.3)d
       20,000                                          A            A            A
                    Webster   52.0(50.7)      29.9(70.7)a   22.6(59.3)°   ND  (47.1)a   24.2(28.1)    16.1  (ND)



    ^irst column, technical  grade  atrazine.

     Second column, formulated atrazine.

    cSignificant at p  <  0.05.

     Significant at p  <  0.01                                  .                          .    (continued)

-------
TABLE 8. (continued)
Concentration
of atrazine-                                       cfu-g"1 soil  (x  10~6)
Cvg'g"1)                                               Time  (Weeks)
                              6             7             8            9            10           11


                Cecil      ND (15.7)     15.2(16.0)   13.9 (ND)     17.7(13.5)    16.3(13.7)    18.5(16.2)
        0
                Webster   13.5(23.4)     18.2(22.8)   16.1(18.5)    19.2(20.0)    20.0(21.1)    19.5(25.2)

                Cecil      ND (20.9)     12.1(14.7)   17.7 (ND)     10.9C(11.4)   12.0(14.8)    13.7(13.9)
       10
                Webster   18.5(33.8)     27.0(16.4)   20.0(19.8)    26.8(18.7)    24.4(21.6)    19.8(19.5)

                Cecil      ND (11.6)     11.4(15.0)   17.6 (ND)     13.7(11.9)    11.9(16.9)    17.1(13.0)
    1,000
                Webster   21.8(19.8)     19.0(19.2)   21.1(27.3)    13.8(23.1)    23.9(21.8)    19.2(27.0)

                Cecil     ND  (49.6)d     13.5(56.3)d  13.9 (ND)     19.8(52.0)d   16.0(36.3)d  19.2(57.2)d
    20,000
                Webster   17.6(30.4)     18.7(28.9)   20.8(32.2)    15.3(23.4)    19.5(25.1)    17.4(23.1)



  rirst column,  technical grade  atrazine.

  Second column, formulated atrazine.

Significant at p <  0.05.

  Significant at p <  0.01                                                            (continued)

-------
    TABLE 8.  (continued)
OO

Concentration
o£ atrazine
Cvg-g"1)
0
10
1,000
20,000
•ill* « 	 •••Him 	 • 	 . — mm 	 - -IP— «l • 	 •!••
cfu-g'1 soil (x 10 ~6)
Time (Weeks)
Cecil
Webster
Cecil
Webster
Cecil
Webster
Cecil
Webster
Average
17.3(17.3)
23.0(26.5)
15.7(17.9)
27.9(26.9)
16.0(17.4)
25.3(26.2)
18.7(67.8)d
23.1(38.1)
•MMmlVIV^V.^l^HIMvmllKBA^^^B^^MM'
           column, technical grade atrazine.
     Second column, formulated atrazine.
    Significant at p < 0.05.
    Significant at p < 0.01.

-------
VO
     TABLE 9.   EFFECT OF TECHNICAL GRADE AND FORMULATED ATRAZINE ON FUNGAL POPULATIONS  IN CECIL AND
               WEBSTER SOILS.



     Concentration
     of atrazine                                        cfu-g"1 soil  (x 10"1*)
     (pg'g"1)                                                Time (Weeks)
                                  0.1            1             234
Cecil
0
Webster
Cecil
10
Webster
Cecil
1,000
Webster
Cecil
20,000
Webster
8.0a(8.

15.2(15
8.9(8.
15.9(16
9.4(7.
17.7(10
7.7(3.

17.3(12
6)b

.2)
6)
-3)
9)
-9)d
8)d

.8)°
8.

14.
10.
15.
9.
14.
8.

16.
1(9.1)

0(15.3)
2(8.5)
3(12.7)
6(9.7)
8(14.4)
8(6. 5)d

5(15.3)
9.

8.
9.
14.
8.
14.
9.

11.
0(8.9)

7(12.1)
5(8.9)
4d(12.1)
4(8.4)
2d(13.3)
1(5. 3)d

4C(14.0)
9.0(8.8)

ND (13.3)
10.0(9.1)
ND (12.9)
7.7(9.5)
ND (14.7)
9.3(6.2)d

ND (14.8)
8.

8.
9.
9.
8.
13.
8.

14.
7 (ND)

4(12.2)
6 (ND)
8(15.0)
8 (ND)
3d(17.8)
9 (ND)
1
6a(12.9)
8

7
8
14
9
12
9

14
.9(8.5)

.0 (ND)
.6(9.4)
.4d(ND)
.5(8.9)
.7d(ND)
.7(6. 5)d
•t
.6a(ND)

            column, technical grade atrazine.

      Second column, formulated atrazine.

     Significant at p < 0.05.

      Significant at p < 0.01.                                                         (continued)

-------
TABLE 9.  (continued)
Concentration
of atrazine-
(yg-g"1)
0
10
1,000
20,000

Cecil
Webster
Cecil
Webster
Cecil
Webster
Cecil
Webster
6
ND (9.
6.1(10
ND (9.
9.9d(10
ND (9.
12.8d(ll
ND (5.
13.8d(12
cfu-g l soil (x
Time (Weeks)
7 8
1)
.6)
1)
.8)
7)
.4)
5)d
.4)
8.
6.
8.
12.
8.
13.
8.
13.
2(8.6)
9(13.1)
4(9.1)
Od(10.2)
8(9.2)
3d(14.7)
7(6. 6)d
5d(11.5)
8.
8.
10.
10.
9.
11.
9.
5 (ND)
1(10.2)
8 (ND)
8(11.9)
4 (ND)
6d(14.8)d
10"^)
9
9.1(9.0)
6.5(11.6)
8.8(9.6)
14.2d(12.1)
8.6(8.4)
9.8d(16.4)d
9 (ND) 7.2(9.4)
9C(14.3)d13.7d(13.1)
10
8.6(8.0)
7.1(11.1)
8.9(9.3)
13.3d(11.0)
7.9(8.6)
11.8d(ll.l)
8.0(8.3)
13.3d(15.0)

9
6
8
12
9
10
7
d!3.
11
.3(9.5)
.5(12.7)
.8(8.6)
.5d(10.4)
.0(8.2)
.6d(14.6)
.6(7.5)
ld(16.0)C

       colijmn, technical grade atrazine.
 Second column, formulated atrazine.
Significant at p < 0.05.
 Significant at p < 0.01.                                                              (continued)

-------
   TABLE 9.  (continued)
ui

Concentration
of atrazine
(yg-g"1)


0


10


1,000


20,000


cfu-g"1 soil (x
Time (Weeks)

Cecil

Webster
Cecil

Webster
Cecil

Webster
Cecil

Webster

10"")

Average
8.7(8.8)

8.6(12.5)
9.3(9.0)
A
13.0a(12.3)
8.8(8.9)
A
13.0 (14.0)
8.6(6.6)d
A
14.0a(13.8)

    Thirst column, technical grade atrazine.



     Second column, formulated atrazine.



    Significant at p < 0.05.



    Significant at p < 0.01.

-------
TABLE 10.  EFFECT OF TECHNICAL GRADE AND FORMULATED ATRAZINE ON ACTINOMYCETE POPULATIONS  IN CECIL
           AND WEBSTER SOILS



Concentrations
of atrazine                                        cfu-g"1 soil  (x 10~5)
(wg-g x)                                               Tijne  (Weeks)
                              0.1             1             2345


                Cecil      4.6a(1.8)b       2.3(3.4)     5.5(10.0)    6.6(10.0)   10.7  (ND)       5.9(6.2)
                Webster    14.6(17.7)      14.6(12.7)     8.8(7.5)     ND  (12.7)    9.6(12.5)     14.0  (ND)


       10
Cecil      5.2(3.0)       7.3(2.5)    9.8C(4.3)C    9.4(7.5)   11.7 (ND)      3.7(7.3)
                Webster    17.4(16.6)      16.4(8.3)    11.2(8.1)      ND (13.0)   12.2(8.8)      12.5 (ND)

                Cecil      6.0(1.6)        6.4(1.4)     7.5(3.9)C     6.4(5.2)c    7.4  (ND)       6.4(5.5)
    1,000
                Webster    19.8(10.4)°     14.8(12.7)    8.8(10.1)     ND (11.4)   13.5(10.4)      8.6C(ND)

                Cecil      3.2(0.2)d       4.8(0.2)d    8.0(1.4)d     11.9d(0.9)d  8.0  (ND)       7.3(0,5)d
   20,000
                Webster    19.0(13.8)      13.8(10.4)   11.2(10.1)     ND (13.0)   13.5(7.5)c     10.7 (ND)



       column, technical grade atrazine.

 Second column, formulated atrazine.

Significant at p < 0.05.

 Significant at p < 0.01.                                                        (continued)

-------
     TABLE 10.   (continued)
in

Concentration
of atrazine
(yg-g'1)
Cecil
0
Webster
Cecil
10
Webster
Cecil
1,000
Webster
Cecil
20,000
Webster

ND

6
(8.4)

ND(10.4)
ND

ND
ND

ND
ND

ND
(7.8)

(10.4)
(11.2)

(7.0)
(0.7)d

(9.6)

1.

12.
2.

8.
2.

13.
3.

15.
d
7
8(5.5)

7(8.5)
7(5.7)

8(7.3)
1(3.0)

3(6.5)
0(1. 4)d

6(6.8)
Pu-g'1 soil (x 10"
Time (Weeks)
8
4.8

13.0
5.2

9.1
5.5

12.5
8.4C

11.7
(ND)

(ND)
(ND)

(ND)
(ND)

(ND)
(ND)

(ND)
1.

9.
1.

10.
2.

14.
2.

12.
5)
9
4(2.7)

9(10.4)
1(2.1)

1(8.1)
1(2.3)

8C(13.8)
3(0. 2)d

2(9.9)
10
3.2(4.3)

ND (11.2)
1.4(4.8)

ND (9.6)
1.1(4.3)

ND (11.4)
2.1(0.5)d

ND (9.1)
11
1.

12.
1.

14.
1.

9.
0.

17.
4(2.1)

0(9.6)
8(0.7)

3(11.4)
4(0.9)

6(11.7)
7(0. 2)c

7(13.5)

            column, technical grade atrazine.



      Second column, formulated atrazine.



     Significant at p < 0.05.



     ^Significant at p < 0.01.
(continued)

-------
    TABLE 10.  (continued)
01
Concentration
of atrazine
(yg-g"1)
0
10
1,000
20,000
cfu'g'1 soil (x 10 "!
Time (Weeks)
Cecil
Webster
Cecil
Webster
Cecil
Webster
Cecil
Webster
5)
Average
4.4(5.4)
12.1(11.3)
5.4(5.2)
12.4(10.2)
4.7(3.8)
12.9(10.5)
5.4(0.6)d
13.9(10.4)

           column, technical grade atrazine.
     Second column, formulated atrazine.
    Significant at p < 0.05.
    Significant at p < 0.01.

-------
TABLE 11.  TOTAL C02 EVOLUTION FROM METHYL PARATHION TREATED WEBSTER SOIL

Time
(day)
2
7
12.5
18.5
25.5
32.5
39.5
47.5

0 ppm
1.81
1.32
1.15
0.86
0.79
0.72
0.51
0.56

24.5
Technical
2.88
1.53
1.25
1.00
0.89
0.82
0.58
0.56
C02-C mg/lOOg
ppm
Formulated
2.40
1.85
0.91
1.24
1.07
0.89
0.58
0.72
soil/day
10,015
Technical
1.99
1.45
1.00
0.93
0.89
0.72
0.54
0.50

ppm
Formulated
2.64
1.73
1.25
0.89
0.82
0.75
0.40
0.61
 Total        45.65      54.94         58.18          47.74         52.79
                                     55

-------
TABLE 12.  TOTAL C02 EVOLUTION FROM METHYL PARATHION TREATED CECIL SOIL

C02-C mg/lOOg soil/day
Time
(day)
2
7
12.5
18.5
25.5
32.5
39.5
47.5
0 ppm
1.93
1.20
0.86
0.82
0.75
0.65
0.61
0.56
24.5
Technical
1.75
1.12
0.86
0.82
0.61
0.65
0.68
0.40
ppm
Formulated
2.16
1.28
0.86
0.72
0.79
0.65
0.79
0.42
10
Technical
1.34
0.55
0.61
0.37
0.54
0.54
0.58
0.40
,015 ppm
Formulated
0.45
0.67
0.57
0.40
0.51
0.47
0.65
0.40'
Total      44.10      40.95         45.04          29.04         26.74
                                  56

-------
TABLE 13.  PERCENT OF  ll*C -METHYL PARATHION EVOLVED AS  14C02  IN WEBSTER AND
           CECIL SOILS


Concentration
(ppm)


24.5

10,015



24.5

10,015







Technical
Formulated
Technical
Formulated


Technical
Formulated
Technical
Formulated



4


45.9
44.8
0
0


36.7
32.6
0
0

\

10


67.0
64.5
0
0


59.7
55.0
0
0

\ evolved as lt*C02
Time (days)
15 29 43

rVfcJDo LCI s-jUjLjL""--"---- — — - —
73.3 80.6 83.8
69.7 76.2 79.2
0 ~0 <0.1
0 ~0 <0.1

— Lecu sou. 	
66.6 75.1 78.5
61.9 70.6 73.9
0 ~0 ~0
0 ~0 <0.1



52


85.0
80.3
<0.1
<0.1


79.4
75.0
~0
<0.1
                                       57

-------
exhibited the characteristics of a bound residue after 28 days, and only 7%
of the 14C-activity was extractable.

     For the 10,015 ppm concentrations, very little technical grade or
formulated methyl parathion was degraded in Webster or Cecil soils during
52 days of incubation.  Less than 0.1% of 14C-methyl parathion was degraded
to 1'»C02 at the 10,015 ppm concentration (technical grade and formulated).
Total C02 evolution from the soils containing 10,015 ppm methyl parathion
was in good agreement with lkC02 evolution.  The total amount of methyl
parathion-carbon added to 100 g of soil at 10,015 ppm was 365 mg.  If
extensive degradation occurred, a substantial amount of C02 would have
evolved.  For example, if 101 of the methyl parathion had mineralized, 36.6
mg of C02-carbon would have evolved.  Total C02 evolution from the Cecil
soil receiving 10,015 ppm of methyl parathion was less than the untreated
soil; therefore, it would not be expected that extensive degradation oc-
curred.  Total C02 evolution for the Webster soil receiving 10,015 ppm of
methyl parathion was not enhanced, and extensive degradation did not occur.
Microbial populations have been reported to be inhibited five years after
parathion application of 30,000 to 95,000 ppm to a soil (Wolfe, et al.,
1973}.

     This project has shown that for low application rates, methyl parathion
was nonpersistent in soils, but the insecticide persisted following appli-
cations of large quantities.  Thus, using the results from the low appli-
cation rates to predict the behavior of methyl parathion at high applica-
tions would lead to erroneous conclusions.

TRIFLURALIN

     Soil respiration rates from Cecil and Webster soils receiving 10 ppm
of technical grade and formulated trifluralin were not different from un-
treated soils except during the first week of incubation (Figures 18 and
19).  Carbon dioxide evolution from the Webster soil receiving 1,000 and
20,000 ppm of technical grade trifluralin were not enhanced except during
the initial incubation period (Figure 19).   Soil respiration from the Cecil
soil treated with 1,000 and 20,000 ppm at technical grade trifluralin was
enhanced during the first two weeks of incubation.  C02 evolution from the
Cecil and Webster soil receiving 1,000 ppm of formulated trifluralin was
also enhanced during the initial period of incubation, and the enhancement
of C02 evolution was greatest for the Webster soil.  C02 evolution in the
Cecil and Webster soils treated with 20,000 ppm of formulated trifluralin
was stimulated in the first 5 days of incubation.  This stimulation was
followed by a later increase in C02 production. Stimulation in C02 produc-
tion in the Webster soil treated with 20,000 ppm of formulated trifluralin
was much greater than that in the Cecil soil receiving the same treatment,
and C02 production' in the Webster soil remained 2 to 9 times higher than
for the untreated Webster soil during the entire 83 days of incubation.

     Less than 5% of the trifluralin applied to the Cecil and Webster soils
was degraded to ltfC02 after 83 days of incubation.  Several metabolites
from the trifluralin were detected and characterized.  Information regarding
metabolites will be presented in the metabolite section of this 'manuscript.


                                      58

-------
Ul
vo
     0 ppm
    1 O ppm
 1,000 ppm
20,000 ppm
        0
                                                     8
                                                           B
                                                      ,
           o—o      0 PPm
           •—•     10ppm
           o—n  1,000 ppm
           •—• 20.OOO ppm
20
                                        40
                                    Days
60
     Figure 18.  C02 evolution rate from the Cecil soil receiving various concentrations of trifluralin
               (A) Technical grade;  (B) Formulated trifluralin.

-------
0\
o
     O
     (ft
     D)
o
 C\J
O
U
                                      Oppm
                                     10 ppm
                                   1,000 ppm
                                 2O.OOO ppm
                   20
                        40
                     Days
60
                                              8
                                                     B
                                                                         o—o
                                  I     I     I
                                           Oppm
                                          10 ppm
                                  a—a  1,OOO ppm
                                                                        20,000 ppm
0
20
    40
Days
60
    Figure 19.   C02 evolution rate from the Webster soil receiving various concentrations of trifluralin
               (A) Technical grade; (B) Formulated trifluralin.

-------
     Bacterial populations in the Webster soil  treated with 10 ppm tri-
fluralin (technical grade and formulated) were  not  significantly different
from the untreated soil  (Table  14).  However, bacterial populations in the
Webster soil receiving 1,000 ppm of  formulated  trifluralin were consistently
higher than the untreated soil  with  the  greater stimulation occurring in
the Cecil rather than the Webster soil  (Table 14).  For 20,000 ppm of
technical grade trifluralin, bacterial populations  in the Cecil soil were
greater than those in the Webster soil.  Bacterial  populations in the Cecil
soil receiving 20,000 ppm of formulated  trifluralin were initially inhibited,
but were enhanced significantly after two weeks of  incubation; whereas
bacterial populations in the Webster soil for the same treatment were
stimulated during the first 18  hours and the stimulation was greatest
during the third week of incubation.

     Fungal populations  in the  Cecil and Webster soils treated with 10 ppm
of technical grade and formulated trifluralin were  generally not signifi-
cantly different from the untreated  soil (Table 15).  For the 1,000 ppm
treatments, fungal populations  in the Cecil soil receiving technical grade
and in the Webster soil  receiving formulated trifluralin were not signifi-
cantly affected, whereas the Cecil soil  treated with the formulated material
and the Webster soil with the technical  grade material showed a significant
effect at the 0.01 and 0.05 levels,  respectively.   Fungal populations were
significantly reduced in both soils  at 20,000 ppm of formulated trifluralin;
whereas fungal populations in the Webster soil  receiving the technical
grade material were stimulated  during the first four weeks.

     Actinomycete populations in Cecil and Webster  soils receiving 10,
1,000 and 20,000 ppm of  trifluralin. (technical  grade or formulation) were
not significantly different from the untreated  soils (Table 16) except in
the 20,000 ppm formulated trifluralin treatments.   Actinomycete populations
in both soils receiving  20,000  ppm of the formulated material were reduced
significantly.

2,4-D

     The rates of total  C02 evolution from the  Webster soil receiving
various concentrations of 2,4-D are  given in Figure 20.  A common character-
istic of the C02 evolution rates for 5,000 and  20,000 ppm was the presence
of two response peaks.   The initial  increase in C02 evolution rate was
followed by a decline and a second increase.  This  was observed for both
forms of 2,4-D.  The C02-carbon (C02-C)  produced prior to the appearance of
the second peak was not  from the 2,4-D-carbon  (2,4-C) because very little
lkCQ2 was measured during this  initial time period.  For formulated 2,4-D,
the C02-C in the first peak appeared to  come from the oxidation of the
formulation ingredients  including dimethylamine; and for technical grade
2,4-D, the C02-C probably originated from impurities and soil organic
matter.  The appearance  of the  second peak coincided with the occurrence
of 2,4-D degradation  (14C-C02 activity)  (Figure 21).  Assuming all C02-C
was evolved from 2,4-D-carbon,  the extent of degradation during the entire
experimental period (80  days) and during the second peak period were
calculated and are shown in Table 17.  These results clearly illustrated
that the majority of the C02-C  evolved during the second peak period was


                                     61

-------
     TABLE 14.   EFFECT OF TECHNICAL GRADE AND FORMULATED TRIFLURALIN ON BACTERIAL POPULATIONS IN CECIL AND
                WEBSTER SOILS


     Concentration
     of trifluralin                                     cfu-g"1 soil (x 10~6)
     (yg-g J)                                                Time (Weeks)
                                  0.1            1             2345


                     Cecil   17.2a(17.0)b   8.7(19.9)     11.4(16.5)    ND (14.4)   16.2(14.9)     ND (12.8)
             0
                     Webster  39.4(25.2)   21.1(17.0)     18.9(15.9)   20.5(15.5)   15.3(14.4)     ND (13.1)

                     Cecil    20.1(16.4)   12.5(17.1)     12.8(14.9)   10.2(15.4)   11.4(16.0)     ND (13.3)
            10                                  ,              .
^                    Webster 46.8(46.5)°   40.0a(28.3)c   31.5a(20.8)   20.7(19.0)   17.6(14.6)     ND (15.0)
tsi
                     Cecil   20.7(40.7)d  11. 5(259. 0)d   10. 7(238. 0)d 11. 1(331. 0)d  9.1(288.0)d   ND(246.0)d
         1,000                                        ,              A           A             A            A
                     Webster 49..1(38.2)c  29. 6(298. 0)a   26. 4(107. 0)a 23.9(42.8)a   17.9(41.3)a   ND (31.2)a

                     Cecil   27.9d(3.6)d  18.5d(70.3)d  33.3d(173.0)d 21. 0(120. 0)d 21.1(74.6)d    ND (62.2)d
        20,000                           ,            ,             -,             •,             A           A
                     Webster  39.9(63.2)a 28. 3(352. 0)a  27. 0(559. 0)a 19. 0(2030. 0)a 18. 6(1140. 0)a  ND(484.0)a
            column,  technical grade trifluralin.

     Second column, formulated trifluralin.

     Significant, at p < 0.05.
    *d         "' •"->
     .significant at p < 0.01.                                                        (continued)

-------
    TABLE  14.   (continued)
ON

Concentration
of trifluralin
Cecil
0
Webster
Cecil
10
Webster
Cecil
1,000
Webster
Cecil
20 000

Webster
6
8.6(16

15.7(13
8.3(14

18.3(14
11.0(284
18.9(36.
26.0d(62


17.0(204

.7)

• 9)
• 1)

.4)
,0)d
?)d
• 8)d

A
.0)d
7
11.3

ND
11.9

ND
10.5
cfu-g"1 soil (x 10"6)
Time (Weeks)
8 9
(ND)

(17.9)
(ND)

(20.3)
(ND)
ND(36.4)d
23.7


d(ND)

A
ND(189.0)"
12.5(12.

16.3(14.
10.6(14.

18.5(18.
11.9(160.
18.2(32.
27.7d(37.


15.3(195.
3)

8)
0)

9)
o)d
Dd
7)d

A
0)d
ND (18.

15.3(12.
ND (15.

17.4(15.
ND(141.
19.1(36.
ND(55.1


16.1(147
1)

7)
0)

3)
o)d
6)d
)d

A
.0)d
10
11

16
15

16
11
19
30


14
.4 (ND)

.6(14.0)
.7 (ND)

.5(15.7)
.3 (ND) 9
.4(32. 6)d
.6d(ND)

j
.4(168. 0)a
11
12.4(14.4)

ND (ND)
14.1(11.5)

ND (ND)
. 0(134. 0)d
ND (ND)
31.0(13.1)


ND (ND)

     aFirst column, technical grade trifluralin.



      Second column, formulated trifluralin.



     Significant at p < 0.05.



     Significant at p < 0.01.
(continued)

-------
TABLE 14.   (continued)
Concentration
of trifluralin
cfu-g"1 soil (x 10"
    Time (Weeks)
       column, technical grade trifluralin.

 Second column, formulated trifluralin.

cSignificant at p < 0.05.

Significant at p < 0.01.
                                                           Average

0


10


1,000


20,000

Cecil

Webster
Cecil

Webster
Cecil

Webster
Cecil

Webster
12.2(15.7)

19.9(15.8)
12.8(15.8)

25.3(20.8)
11. 7(212. 0)d
A
25.5(66.6)a
26.1d(67.2)d
A
21. 7(503. 0)a


-------
    TABLE 15.   EFFECT OF TECHNICAL GRADE AND FORMULATED TRIFLURALIN ON FUNGAL POPULATIONS IN CECIL AND
               WEBSTER SOILS
ON
tn

Concentration
of trifluralin
G-ig'g"1)


0


10


1,000

20,000

Cecil

Webster
Cecil
Webster

Cecil
Webster
Cecil
Webster

12.2

14.
13.
16.

13.
14.
13.
13.
0.1
a(14.3)b

0(13.4)
3(14.5)
0(14.6)

8(11.0)
0(12.1)
7(6. 9)d
7(12.6)

9.

9.
13.
16.

11.
14.
10.
15.
1
1(12.7)

3(11.7)
1(10.1)
ld(12.7)
A
4(6. 9)a
Od(12.3)
3(0. 7)d
Od(3.4)d
cfu-g"1 soil (x 10"1*)
Time (Weeks)
2
10.5(13.

9.4(10.
12.2(11.
13.5d(ll.

10.9(13.
15.0d(13.
9.7(0.2)
14.2d(2.1

6)

5)
3)
5)

1)
7)
d
)d
3
11.4(11.7)

10.8(12.0)
10.7(12.4)
13.9(13.8)
A
10. 5(6. 0)d
13.8(13.6)
9.3(0.3)d
13.3(0.3)d
4
10.6(10.3)

8.6(12.5)
10.4(11.1)
14.4d(12.3)
A A
6.1d(6.2)d
13.8d(13.6)
6.5d(0.2)d
13.3d(0.4)d
5
ND

ND
ND
ND

ND
ND
(10.7)

(10.3)
(10.5)
(12.4)
A
(7.9)d
(10.9)
ND(<0.1)d
ND
(0.3)d

     Tarst  column,  technical grade trifluralin.


      Second column,  formulated trifluralin.


     Significant  at p  <  0.05.


     Significant  at p  <  0.01.
(continued)

-------
     TABLE 15.   (continued)
o\

Concentration
of trifluralin
(ug-g"1)

Cecil
0
Webster
Cecil
10
Webster
Cecil
1,000
Webster
Cecil
?n nnn
L\) , UUU
Webster
cfu-g l soil (x 10 **)
Time (Weeks)

9

9
6

13
11

14
9


11
6
.8(8.

.5(12

8)

• 3)
.2d(10.4)

.7(9.
.9(6.

.3(13
.7(0.


.2(0.

6)
8)C

•8)
2)d

A
3)d
7
11.2

ND
11.5

ND
11.7

ND
9.2



(ND)

(11.2)
(ND)

(15.1)
(ND)

(13.4)
(ND)

A
ND(0.3)"

11.

9.
19.

12.
12.

12.
11.


10.
8
1(9.9)

6(10.0)
7d(10.3)

4(12.8)
7(5. 8)d

2(15.1)
5(0. l)d

A
3(0. 2)a
.9
ND (13.


1)

9.7(8.9)
ND (16.

11.3(10.
ND(6.3)

12.7(15.
ND(<0.1


10.9(<0.
4)

3)
d
A
0)d
1d

A
1 *\
10
9.9 (ND)

9.2(10.7)
11.4 (ND)

12.9(11.1)
8.9 (ND)
A
14.0(12.9)a
9.0 (ND)

A
10.7(<0.1)a
11
8.9(12.0)

ND (ND)
9.9(8.1)d

ND (ND)
19.7(6.3)d

ND (ND)
9.3(0.0)d


ND (ND)

           column,  technical  grade trifluralin.
     Second column,  formulated trifluralin.
    Significant  at  p  <  0.05.
     Significant  at  p  <  0.01.
(continued)

-------
TABLE 15.   (continued)
Concentration
of trifluralin
(yg'g *)
cfu-g"1 soil (x 10"4)
    Time (Weeks)
 aFirst column, technical grade trifluralin.

  Second column, formulated trifluralin.

 Significant at p < 0.05.

 ^Significant at p < 0.01.
                                                           Average

0


10


1,000
7

20,000

Cecil

Webster
Cecil

Webster
Cecil

Webster
Cecil

Webster
10.5(11.7)

10.0(11.2)
11.8(11.5)

13.8C(12.4)
11.8(7.6)d

13.8C(13.3)
9.8(0.9)d
_ j
12.5c(1.8)a


-------
    TABLE 16.  EFFECT OF TECHNICAL GRADE AND FORMULATED TRIFLURALIN ON ACTINOMYCETE POPULATIONS  IN CECIL
               AND WEBSTER SOILS
oo

Concentration
of trifluralin
(ug-g"1)
0
10
1,000
20,000

Cecil
Webster
Cecil
Webster
Cecil
Webster
Cecil
Webster

5
14
5
27.
12
16
7
15
0.1
.2a(11.4)b
.0(11.2)
.2(14.3)
8C(12.5)
.ld(9.8)
.9(13.0)
.1(15.3)
.9(6. 0)C
1
6.8(9.8)
12.0(13.3)
5.7(6.8)
13.5(14.3)
4.6(3.2)d
16.6(13.5)
6.4(7.3)
14.0(2.1)d
cfu-
5
18
6
15
7
13
5
13
g l soil (x
Time (Weeks)
2
.2(8.9)
.5(8.8)
.2(4.6)
.6(12.7)
.5(6.2)
.3(13.3)
.2(1.9)d
.3(1.3)d
10 ~5)
3
ND (7.5)
16.4(18.2)
ND (6.2)
15.1(15.6)
ND (3.4)
15.1(16.1)
ND (l.l)d
15.1(0.7)d

4
11
5
14
5
13
4
14
4
.6(5.0)
.4(14.6)
.5(4.8)
.0(15.6)
.1(5.0)
.3(16.6)
.8(1. 4)d
.0(0.3)d
5
ND
ND
ND
ND
ND
ND
ND
ND

(ND)
(14.6)
(ND)
(13.3)
(ND)
(14.0)
(ND)
(0.2)d

           column, technical grade trifluralin.
     Second column, formulated trifluralin.
    Significant at p < 0.05.
    Significant at p < 0.01.
(continued)

-------
    TABLE 16.  (continued)
Os
	 . 	 .
Concentration
of trifluralin
Cng'g'1)
0
10
1,000
20,000

Cecil
Webster
Cecil
Webster
Cecil
Webster
Cecil
Webster

4.
14.
2.
14.
3.
14.
5.
15.
6
1(7.8)
2(10.9)
5(6.6)
7(12.0)
9(6.6)
0(16.1)
2(1.8)d
0(0. 2)d
7
4.1
ND
3.4
ND
5.0
ND
3.9
ND
cfu-g l soil (x
Time (Weeks)
8
(ND)
(15.3)
(ND)
(19.5)
(ND)
(18.2)
(ND)
(0.2)d
5.
16.
4.
15.
6.
11.
3.
14.
0(5.5)
9(9.1)
1(4.1)
9(18. 2)d
2(5.2)
4(12.5)
7(0. 4)d
6(0. 2)d
10 ~5)
9
ND (5.7)
11.2(9.1)
ND (5.0)
10.7(10.9)
ND (7.1)
15.1(10.9)
ND(<0.1)d
10.4(0.0)d

5
12
4
13
4
14
3
19
10
.9 (ND)
.2(10.9)
.6 (ND)
.3(13.5)
.8 (ND)
.0(17.9)
.4 (ND)
.2(0. 0)d
11
3.4(7.5)
ND (ND)
3.2(4.8)
ND (ND)
3.7(4.6)
ND (ND)
4.6(0.0)d
ND (ND)

     aFirst column,  technical grade trifluralin.



      Second column, formulated trifluralin.



     Significant at p < 0.05.



      Significant at p < 0.01.                                                       (continued)

-------
TABLE 16.   (continued)

Concentration
of trifluralin
Cvg-g"1)
0
10
1,000
20,000
cfu-g l soil (x
Time (Weeks)
Cecil
Webster
Cecil
Webster
Cecil
Webster
Cecil
Webster
10"5)
Average
4.9(7.7)
14.1(12.4)
3.2(6.4)
15.6(14.4)
3.7(5.7)
14.4(14.7)
4.9(3.2)C
14.6(1.0)d

       column, technical grade trifluralin.
 Second column, formulated trifluralin.
Significant at p < 0.05.
Significant at p < 0.01.

-------
         30
         20
      o
      o"
      O
      o»
          10
                                            o—o
                                   A
      I
                                  Days
                                      Oppm
                                      SOppm
                                      SOOppm
                                      5,000ppm
                                      20,000ppm
                                      Oppm
                                      SOppm
                                      SOOppm
                                      5,000 ppm
                                      ZO.OOOppm
                                                 60
                                                  80
Figure 20.
C02 evolution rate from the Webster soil receiving various con-
centrations of 2,4-D.  (A) Technical grade;  (B) Formulated 2,4-D.
                                    71

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from the carbon in the 2,4-D.  The C02 evolution rates were generally
higher from the formulated 2,4-D treated soil than from the technical grade
treated soil.  Therefore, microbial activity in the soil with formulated
treatments was higher.

     Unlike the Webster soil, total C02 evolution was inhibited in the
Cecil soil which received 5,000 and 20,000 ppm of technical grade 2,4-D as
well as 20,000 ppm of formulated 2,4-D (Figure 22).  Small stimulations
were noted in the total C02 evolution from the soil treated with 5,000 ppm
of formulated 2,4-D.  In contrast to the Webster soil, very little degrada-
tion was observed in the Cecil soil receiving 5,000 and 20,000 ppm (Figure
23). At a rate of 500 ppm, both forms of 2,4-D were degraded slowly.  Only
10.5% and 6.3% of the technical and formulated material were degraded, re-
spectively, in the Cecil soil after 80 days of incubation.

     The C02 evolution rates from the Terra Ceia soil receiving 2,4-D are
presented in Figure 24.  The C02 evolution rate from the untreated organic
soil was approximately four times higher than that from the Webster soil
and 11 times higher than that from Cecil soil.  Total C02 evolution from
the organic soil treated with 5,000 and 20,000 ppm of formulated 2,4-D was
high. For the technical grade material, C02 evolution was inhibited initially.
The C02-C produced from 5,000 ppm of formulated 2,4-D after 7 days was at
least partly from the 2,4-D-C because significant degradation occurred
thereafter.  For the organic soil treated with 20,000 ppm of formulated
2,4-D, total C02 evolution was enhanced after 36 days.  This increase in
C02 evolution was concurrent with the degradation of the herbicide.  The
C02 evolution rate for the organic soil treated with 5,000 ppm of the
technical grade 2,4-D increased after 20 days of incubation.  This increase
coincided with the period of rapid 2,4-D degradation (Figure 25).  Total
C02 evolution was inhibited throughout the 80 days of incubation for the
organic soil treated with 20,000 ppm of technical grade 2,4-D.  Similar to
the Webster soil, comparable degradation results were obtained for the
organic soil when total C02 evolution during the second peak rather than
the entire experimental period was used to calculate 2,4-D degradation.

     Degradation rates for the formulated 2,4-D in the Webster and organic
soil were greater than for the technical grade material when the applica-
tion rate was 5,000 ppm or higher.  Table 18 shows the rates of first order
(exponential) degradation for the three soils receiving 5,000 ppm of 2,4-D.
Exponential degradation was not observed for the Cecil soil.

     Attempts were made to stimulate 2,4-D degradation in the Cecil soil
receiving 5,000 ppm of 2,4-D.  In addition to 2,4-D (technical grade or
formulated) and ^T-2,4-D, the Cecil soil was amended with various nutrient
sources.  Table 19 shows the amount of 2,4-D degradation during 60 days of
incubation.  Readily degradable nutrients such as yeast extract (1%) and
glucose (1%)  plus urea(0.5%) did not stimulate 2,4-D degradation.  Only
a small stimulation, if any, occurred with these treatments with the excep-
tion of the treatment with lime plus 2,4-D degrading bacterium.  In this
treatment 28.6 and 3.91 of the formulated and technical grade 2,4-D,
respectively, were degraded in 60 days.
                                     72

-------
            too
                                            o—o 50 ppm
                                            •_• 500ppm
                                            A—A 5,000 ppm
                                               20,000 ppm
                                             o  .o 50ppm
                                                  SOOppm
                                                  SpOOppm
                                                  20,000 ppm
                                                             80
Figure 21.  Percent 14C02 evolved from the Webster soil receiving various
            concentrations of 2,4-D.  (A) Technical grade; (B) Formulated
            2,4-D.

                                     73

-------
   X.
   o
        0

        3
    8  2
    is
    O
    O
                                 A
20
40
                                      B
                        a—a 0  ppm
                        •—• 500 ppm
                        A—A 5,000 ppm
                             20,000 ppm
60
80
                            o—D 0  ppm
                            •—• 500   ppm
                            £—A 5,000 ppm
                            A—* 20,000 ppm
                     20
              40
             Days
             60
             80
Figure 22.  C02 evolution rate from the Cecil soil receiving various concen-
           trations of 2,4-D.  (A)  Technical grade;  (B) Formulated 2,4-D.
                                   74

-------
         15
         10
       i
       I
A
                                        500 ppm
                                        SpOOppm
                                        20,000 ppm
                       20
 40
                                     60
80
                                 B
               500 ppm
               5000 ppm
               20pOOppm
                                                60
                          80
Figure 23.
                        40
                      DAYS
Percent 14C02  evolved from the Cecil soil receiving various con-
centrations  of 2,4-D.  (A) Technical grade;  (B) Formulated 2,4-D.
                                   75

-------
           15
          >»
          olO
          o
          o
           N
          o»
          2
Formulated           Technical
5000 ppm A—A      5000 ppm  o—o
20000ppm A—A     20,000 ppm •—•
       Control  a—a
                                                          A-A
                        20
             40

           DAYS
60
80
Figure  24.  C02 evolution rate from the Terra Ceia soil receiving 5,000 and
           20,000 ppm of technical grade and formulated 2,4-D.
                                   76

-------
        60
      CO
      O
      o
     u 20
              Formulated
              SOOOppm  A—A
              20pOOppm A—A
                        if  Technical
                      ,     SOOOppm  o-o
                           20,000ppm •-•
          0
20
40
DAYS
                                             60
                                    80
Figure  25.  Percent 14C02 evolved from the Terra Ceia soil receiving 5,000
           and 20,000 ppm o£ technical grade and formulated 2,4-D.
                                   77

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TABLE 17.  COMPARISON OF THE PERCENTAGE OF 2,4-D DEGRADATION FROM THE WEBSTER
           SOIL DERIVED FROM ll*C02 AND TOTAL C02 EVOLUTION DURING THE SECOND
           PEAK PERIOD AND THE TOTAL EXPERIMENTAL PERIOD
Form of 2,4-D
                % degradation        % degradation from
Concentration   calculated from      of ^C-activity
    (ppm)        total C02 evolution  evolved as 14C02

Technical grade


Formulated

5,000

20,000
5,000

20,000
32.7* (87. 0)**

14.2 (18.2)
77.3 (152.0)

21.4 (52.1)
30.3* (56.9)**

11.6 (12.9)
55.5 (60.4)
,
21.0 (22.7)

*The second peak period

**80-day period
                                      78

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TABLE 18.  DEGRADATION RATES FOR THE THREE SOILS RECEIVING 5,000 ppm OF
           TECHNICAL GRADE AND FORMULATED 2,4-D DURING EXPONENTIAL DEGRA-
           DATION PERIOD
                   Exponential     ^degradation (% of    Rate  of degradation
                   degradation     2,4-D evolved as      during this period
                   period (day)         C02-C)                (% per  day)
Webster soil +
technical grade
2,4-D 10th- 45th
Webster soil +
formulated 2,4-D 19th-42nd
Organic soil +
technical grade
2,4-D 18th-42nd
Organic soil +
formulated 2,4-D 7th- 26th
Cecil soil +
technical grade
2,4-D none
Cecil soil +
frvnmil a-f-Arl 7 A-Tt none

43.1

53.9
32.4
59.0

~0
~0

1.23

2.34
1.35
3.11

~0
~0
                                       79

-------
     The soil pH of the Webster and the Cecil soils receiving formulated
2,4-D were not changed appreciably during 11 weeks of incubation  (Tables 20
and 21).  However, the pH of the two soils decreased after receiving  5,000
and 20,000 ppm of technical grade 2,4-D, especially the Cecil soil.   The pH
of the Webster soil receiving 5,000 and 20,000 ppm of technical grade 2,4-D
increased to 7.1 and to 6.7, respectively after the llth week of  incubation.
pH changes were small in the Terra Ceia soil receiving the technical  grade
and formulated 2,4-D.

     At application rates of 5,000 ppm or higher, if 2,4-D degradation
occurred, total C02 evolution was generally enhanced.  In this case,  the
pattern of the 0)2 evolution rate was generally a two peak response.   The
results from 2,4-D degradation indicated that the C02-C from the  second
peak appeared to be associated with the 2,4-D-C (oxidation of 2,4-D-C to
C02-C) and the C02-C from the first peak was probably from formulation
compounds, impurities and/or soil organic matter.  Thus, using total  C02
evolution for the entire experimental period to calculate the extent  of
2,4-D degradation may lead to erroneous conclusions.

     It was observed that if degradation occurred at high 2,4-D application
rates, the degradation rate for formulated 2,4-D was generally higher than
that for the technical material.  This may be attributed to the additional
carbon or nutrients present in the formulation material.  The solubility  of
the dimethylamine salt of 2,4-D in water is 300 g per 100 g water, whereas,
the solubility of 2,4-D is only 900 ppm (Weed Science Society of America,
1974).  Therefore, formulated 2,4-D, even at 20,000 ppm, would remain in
the soil aqueous phase, whereas the majority of technical grade 2,4-D at
concentrations of 5,000 and 20,000 ppm would remain as a solid.  Also  the
2,4-D in the formulation was neutralized by dime thy 1 amine; thus,  the  soil
pH did not change appreciably.  The pH in the soil receiving technical
grade 2,4-D decreased up to 3 pH units, depending upon the soil type  and
2,4-D concentration.  This decrease in pH may have an inhibitory  effect on
soil microbial activity.

     For low application rates, i.e., below 100 ppm, 2,4-D degradation has
been shown to be favored by moisture and organic matter (Foster and McKer-
cher, 1973; Loos, 1969).  Based on the information presented in this manu-
script, herbicide degradation in the presence of high concentrations was
also higher in soils with large quantities of organic matter.  Numerous
other factors may also influence the rate of degradation.  For example,
2,4-D concentration, concentration of formulation material, supplement of
external nutrients, clay, soil pH, arid temperature.  Increasing the herbi-
cide concentration may exhibit direct and indirect effects on microbial
activity.  For example, bacteria have been reported as being the major
organisms responsible for 2,4-D degradation in soils (Loos, 1975), and the
activity of bacteria will be significantly reduced by a low soil pH.   The
buffering capacity of the Webster soil and organic soil was greater than
that for the Cecil soil.  The pH of the Cecil soil dropped 1.9 and 2.7 units
when application rates of technical grade 2,4-D were 5,000 and 20,000 ppm,
respectively; whereas, the pH of the Webster soil declined only 0.6 and
1.2 units for 5,000 and 20,000 ppm, respectively.  Only a small pH change
was observed in the organic soil receiving 5,000 ppm of technical grade

                                      80

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TABLE 19.  2,4-D DEGRADATION  IN THE CECIL SOIL RECEIVING 5,000 ppm OF THE
           HERBICIDE AND VARIOUS NUTRIENT TREATMENTS DURING 60 DAYS OF
           INCUBATION
% Degradation
Treatment Technical Formulated
None
Glucose (1%) + Urea (0.51)
Yeast extract (1%)
Webster soil (21)
Terra Ceia organic soil (2%)
Cow manure (21)
2,4-D degrading bacterium (1 x 106 cells/ g soil)*
Lime (pH 7.5)
Lime (pH 7.5) + Terra Ceia organic soil (2%)
Lime (pH 7.5) + 2,4-D degrading bacterium
(1 x 106 cells/g soil)
0.2
0.2
0.2
0.6
0.3
0.4
0.4
0.9
1.1
3.9

mmimmmm**i^^mm*mmiim-~+immii^^~~~~~~~*~*^*
0.2
0.1
0
0.4
0.4
0.5
0.4
0.6
0.4
28.6

 *2 4-D degrading bacterium was isolated from the Webster soil treated with
 5,000 ppm of technical 2,4-D.   The bacterium was a gram-negative, motile rod.
                                      81

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TABLE 20.  SOIL pH OF THE WEBSTER SOIL RECEIVING TECHNICAL GRADE AND FORMU-
           LATED 2,4-D

pH
Weeks
0 ppm
0 7.3
6 7.3
11 7.2

500 ppm
7.3* (7.3)**
7.3 (7.4)
7.3 (7.4)

5,000 ppm
6.7 (7.3)
7.0 (7.4)
7.1 (7.4)

20,000 ppm
5.1 (7.4)
6.2 (7.4)
6.7 (7.4)

technical grade 2,4-D

**Formulated 2,4-D


TABLE 21.  SOIL pH OF THE CECIL SOIL RECEIVING TECHNICAL GRADE AND FORMULATED
           2,4-D


Weeks

0
6
11
PH

0 ppm
5.6
5.6
5.4

500 ppm
5.3* (5.9)**
6.0 (6.2)
5.8 (5.9)

5,000 ppm
3.9- (5.9)
3.7 (6.1)
3.8 (5.9)

20,000 ppm
3.7 (6.2)
2.9 (5.9)
2.8 (5.8)

*Technical grade 2,4-D

**Formulated 2,4-D
                                     82

-------
 i c Ann    inab^y  o£ the Cecil soil to degrade 2,4-D  at  a  concentration
of 5,000 ppm or higher appeared to be due to concentration effects of the
herbicide and the  formulation chemicals, low PH,  low organic matter content,
and low 2,4-D degrading microorganism populations.

     The bacterial population in the 500 ppm technical grade 2,4-D treated
Cecil soil was significantly lower than that in the untreated  Cecil soil
except during the  6th  and 7th week (Table 22).  During the 6th and 7th week
there was no significant difference at the 0.05 level.  The  bacterial
density in the 500 ppm formulated 2,4-D treated soil was  also  significantly
lower than that of the control (P < 0.05 level),  except during the 2nd
week.  The overall bacterial population in the  500 ppm technical grade
treated soil during  the entire experimental period was 7.99  x  106 cfu per g
soil which was significantly lower (0.05 level) than the  1.11  x 107 cfu
per g soil in the  untreated soil.   Also, the overall average bacterial
density for 500 ppm  formulation treated soil was  significantly lower than
the control, i.e., 6.1 x 106 vs 1.0 x 107.   As  the 2,4-D  concentration was
increased to 5,000 and 20,000 ppm, the bacterial  populations declined pro-
foundly even after three hours of incubation.   Furthermore,  the bacterial
populations in the 5,000 and 20,000 ppm treated soil, for both technical
grade and formulated,  were all significantly lower (P < 0.01)  than from the
untreated soil.  The 20,000 ppm treatments  had  a  more detrimental effect
than 5,000 ppm.  The overall average bacterial  populations for 5,000 and
20,000 ppm technical 2,4-D treated soil were 1.7  x 106 and 7.0 x 105,
whereas the bacterial  populations for 5,000 and 20,000 ppm formulated
treated soil were  1.9  x 106 and 8.2 x 10s,  respectively.

     Both forms of 2,4-D significantly stimulated fungal  population (P <
0.01) when the application was at 500 ppm (Table  23).  The stimulation was
greater for the formulation treatment.   At  an application rate of 5,000
ppm, formulated 2,4-D  did not stimulate or  inhibit fungal population (not
significant at 0.05  level), whereas technical 2,4-D profoundly inhibited
fungal populations throughout the entire experimental period (P < 0.01).  At
20,000 ppm, both forms of 2,4-D exhibited inhibition on fungal populations.
Fungal populations were reduced drastically after 3 hours of incubation,
and completely destroyed in 7 and 6 weeks after incubation with technical
and formulated 2,4-D,  respectively.

     Actinomycete  populations in the 500 ppm technical 2,4-D treated soil
were significantly reduced (P < 0.01),  except during the  5th week of incu-
bation (Table 24).   Whereas, for formulated 2,4-D at the  same  application
rate, actinomycete populations were significantly lower than in the un-
treated soil (P <  0.01)  after the 5th week.   At application  rates of 5,00.0
and 20,000 ppm, both forms of 2,4-D caused  sharp  declines in actinomycete
populations.  At these concentrations,  technical  grade 2,4-D appeared to
have a greater inhibitory effect on actinomycetes in Cecil soil.  Actinomy-
cetes were not detected in the SC agar after 2  weeks of incubation; whereas,
actonomycetes were not detected after 4 and 5 weeks in soils treated with
5,000 and 20,000 ppm of formulated 2,4-D.

     Generally, technical grade 2,4-D had a greater inhibitory effect on
the bacteria/fungi  and actinomycetes in the Cecil soil for  applications
                                     83

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      TABLE 22. EFFECT OF 2,4-D ON BACTERIAL POPULATIONS IN CECIL SOIL

Concentration
of 2,4-D
Cvig'g"1)


0


500


5,000


20,000

cfu'g'
'l soil (x 10 ~6)
Time (Weeks)

Technical

Formulated
Technical

Formulated
Technical

Formulated
Technical

Formulated
0(3h)
12.5

12.3
11.0

5.0*
3.2*

2.1*
0.7*

1.3*
1
10.6

15.0
7 fif
/.O

7.6*
1.1*

1.4*
1.1*

0.8*
2
11.9

8.1
7.3f

7.8
2.5*

1.0*
1.1*

1.1*
3
10.3

11.2
4.4*

8.7f
2.9*

1.7*
0.7*

0.5*
4
11.4

9.0
8.1f

5.6f
1.3*

1.0*
0.5*

0.8*
5
14.0

12.7
n.d.

7.6f
n.d.

3.1*
n.d.

0.7*

*Signi£icant at p < 0.01.
 Significant at p < 0.05.
                                                         (continued)
                                     84

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TABLE 22.   (continued)
Concentration
of 2,4-D                                 cfu-g'1  soil  (x 10"6)
(vg-g *)                                     Time (Weeks)
                             6      7      8      9     10     11     Average

0


500


5,000


20,000

Technical

Formulated
Technical

Formulated
Technical

Formulated
Technical

Formulated
10.9

10.0
10.9

4.6*
1.6*

1.1*
0.3*

0.7*
8.6

11.5
9.5

6.5*
0.7*

3.4*
0.6*

n.d.
8.1

n.d.
6.9f

n.d.
1.4*

n.d.
0.8*

n.d.
12.9

8.9
7.0f

3.2*
1.0*

2.6*
0.5*

0.9*
10.

10.
7.

4.
1.

1.
0.

0.
9

8
3f

1*
5*

3*
5*

8*
n.

9.
n.

6.
n.

2.
n.

0.
d.

4
d.
•
3f
d.

1*
d.

7*
11.

10.
8.

6.
1.

1.
0.

0.
1

8
Of

1*
7*

9*
7*

8*
*Signi£icant  at p <  0.01.

Significant  at p <  0.05.
                                      85

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       TABLE 23.  EFFECT OF 2,4-D ON FUNGAL POPULATIONS IN CECIL SOIL

Concentration
of 2,4-D
(yg-g"1)


0


500
•

5,000


20,000

cfu-g
~l soil
(x 10 "^


Time (Weeks)

Technical

Formulated
Technical

Formulated
Technical

Formulated
Technical

Formulated
0(3h)
25.7

16.3
25.9

14.8
1.9*

4.0*
0.2*

1.6*
1
10.5

6.6
24. 4f

71.8*
0.2*

3.2*
0.2*

<0.1*
2
9.1

10.8
48.5*

124*
0.2*

12.3
0.4*

<0.1*
3
10.2

10.8
57.9*

103*
0.9*

10.3
0.2*

0.4*
4
n.d.

12.5
n.d.

133*
n.d.

11.4
n.d.

<0.1*
5
12.8

13.2
70.9*

144*
0.8*

13.6
0.1*

0.1*

*Signi£icant at p < 0.01.
"^Significant at p < 0.05.
                                                          (continued)
                                     86

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TABLE 23.   (continued)
Concentration
of 2,4-D                                 cfu-g'1  soil  (x  10"1*)
(yg*g *)                                     Time (Weeks)
                             6     7     8     9     10      11     Average


             Technical     9.1   5.7   9.3   3.4   5.2     n.d.     10.1
        0
             Formulated   10.9   9.7   9.8   8.8   7.1   10.5      10.6

             Technical    70.9* 69.0* 77.1*  33.7*   74.3*   n.d.   '  55.3*
      500
             Formulated  132*  324*  310* 196*    204*   192*      162*

             Technical     0.2* <0.1*  0.1*  <0.1*   0.2*   n.d.      0.4*
     5,000
             Formulated   31.0* 13.1  13.0  12.5   6.0     7.9      13.8

             Technical    <0.1*  0*    0*    0*     0*      n.d.      0.1*
   20,000
             Formulated   <0.1* <0.1*  0.1*   0*     ,0*      0*        0.2*
 *Signi£icant at p < 0.01.

 Significant at p < 0.05.
                                      87

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    TABLE 24-  EFFECT OF 2,4-D ON ACTINOMYCETE POPULATIONS IN CECIL SOIL

Concentration
of 2,4-D
(vg-g"1)


0


500


5,000


20,000

cfu-g'
~l soil (x 10 "^
Time (Weeks)

Technical

Formulated
Technical

Formulated
Technical

Formulated
Technical

Formulated -
0(3h)
15.4

2.4
14.1

2.4
8.4*

0.3*
0.1*

0.4*
1
14.8

1.2
2.5*

1.1
<0.1*

0.2*
<0.1*

0.1*
2
n.d.

2.4
n.d.

1.1
n.d.

0.2*
n.d.

<0.1*
3
8.9

2.3
1.3*

3.0
0*

<0.1*
0*

<0.1*
4
n.d.

2.1
n.d.

2.3
n.d.

<0.1*
n.d.

0*
5
1.2

5.0
1.9

0.5*
0*

0*
0*

0*

*Signi£icant at p < 0.01.
                                                     (continued)
                                     88

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TABLE 24.  (continued)
Concentration
of 2,4-D
(yg-g l)

Technical
0
Formulated
Technical
500
Formulated
Technical
5,000
Formulated
Technical
20,000
Formulated


cfu*
g"1 soil (x 10 "•*)
Time (Weeks)
6
1.7

1.1
0.2*

0.2*
0*

0*
0*

0*
7
1.7

1.3
0.2*

0.2*
0*

0*
0*

0*
8
1.4

n.d.
0.2*

0.2*
0*

n.d.
0*

n.d.
9
n.d.

2.6
n.d.

0.2*
n.d.

0*
n.d.

0*
10
1.7

1.7
<0.1*

0.5*
0*

0*
0*

0*

11
n.d.

2.0
n.d.

0.2*
n.d.

0*
n.d.

0*

Average
5.9

2.2
2.6*

1.1*
1.1*

0.1*
0.02*

0.1*
*Significant  at p  <  0.01.
                                       89

-------
of 5,000 ppm and higher.  This may be attributed to the fact that the soil
pH dropped from 5.6 to 3.7 and 2.9 in the soil treated with 5,000 and
20,000 ppm of technical grade 2,4-D.  The pH in the formulated treated
soils remained unchanged.  Also, formulation compounds in the formulated
2,4-D may reduce or buffer 2,4-D toxicity to the microorganisms.  Among the
three groups of microorganisms considered, bacteria were the least sen-
sitive to high 2,4-D concentrations (5,000 ppm and higher) and actinomycetes
were the most sensitive for the soils considered.  However, for the soils
that degraded 2,4-D at high concentrations, for example, Webster silty clay
loam and Terra Ceia muck  (Ou et al., 1978), the response of the soil micro-
flora to high 2,4-D concentrations may be different.  Microorganisms in the
20,000 ppm treated Webster soil after 70 days of incubation were nearly all
bacteria, and the number  of bacteria was more than 100 times higher than
that in the untreated soil.

METABOLITES

Atrazine

     Only tentative indent if icat ions of atrazine metabolites were made.
Structures are based on thin layer and gas chromatographic behavior.  Mass
spectral analyses are currently incomplete.  In the Webster soil treated
with 10 or 1,000 ppm, 4 to 8 percent of the radioactivity was associated
with a compound or compounds having a relative Rf similar to that of 2-
chloro-4-amino-6-isopropylamino-s-triazine (0.80J and/or 2-chloro-4-ethyl-
amino-6-amino-s-triazine  (0.88).  One to four percent of the activity was
associated with a component having a relative R^ indistinguishable from
that of 2-hydroxy-4-ethylamino-6-isopropylamino-s-triazine (0.48; hydroxy-
atrazine) .  Less than 2 percent of the radioactivity was detected at a
relative Rr similar to that of 2-hydroxy-4-amino-6-isopropyl-amino-s-
triazine (0.10) and 2-hydroxy-4-ethylamino-6-amino-s-triazine (0.10). In
the Cecil soil, all the radioactivity not associated with atrazine had a
relative Rr corresponding to hydroxy- atrazine (0.48).  Percentages of
metabolites increased with time and corresponded to a reduction in the
level of the parent compound.

     The percentage of unextractable radioactivity was generally higher at
10 and 1,000 ppm for the  Cecil soil than for the Webster soil; this differ-
ence was not observed at  the 20,000 ppm treatment.  There was no apparent
difference in bound residues between the technical and formulated appli-
cations at any of the three concentrations studied (Figure 26) .  The per-
centage of bound 14C-activity increased with time and as much as 30% of the
    detected in the soil was unextractable.
Trifluralin
     Trifluralin degradation in the Webster and Cecil soils measured by
      evolution from the 14C-labeled trifluralin decreased on a percentage
basis with an increase in herbicide concentration during the 83 days of
aerobic incubation (Table 25) .  Degradation was greater in the Webster soil
for all concentrations (10, 1.000 and 20,000 ppm) than in the Cecil soil.
The percentage volatilized (1(*C-material trapped in ethylene glycol) was
                                      90

-------
              80
                                     WEBSTER TECH
                                     WEBSTER FORM
                                     CECIL TECH.
                                     CECIL  FORM.
                0  10 20  30 40  50 60  70 80

                  INCUBATION TIME (days)
Figure 26.  Percent of total ll*C-activity bound to Webster and Cecil soil
          after receiving 10, 1,000 and 20,000 ppm of technical grade
          and formulated atrazine.
                              91

-------
negligible for all treatments except for the 10 ppm formulated application.
In this treatment, 0.11 of the total 11+C-activity was trapped in the ethylene
glycol in the first three weeks of incubation, but no ^C-activity was
detected thereafter.  It was apparent, however, that volatilization occurred
to a certain extent based on the orange color observed on the tygon tubing
connecting the soil incubation flask to the ethylene glycol test tube.  The
tygon tubing was extracted with acetone and the amount of ltfC-activity
associated with the orange color determined.  The percentage of the total
amount of trifluralin evaporated decreased with an increase in herbicide
concentration, although a greater quantity was volatilized at the higher
concentrations (Table 25).

     The chromatographic and mass spectral characteristics of the metabo-
lites detected are presented in Tables 26 and 27.  Table 27 presents the GC
and TLC behavior and mass spectral fragmentation pattern for each material
isolated.  In addition to the trifluralin, four compounds were identified:
(1), a,a,a-trifluoro-2,6-dinitro-N-propyl-p-toluidine (2), a,a,a-trifluoro-
2,6-dinitro-p-toluidine (3), 2-ethyl-7-nitro-l-propyl-5-trifluoromethyl
benzimidazole (4) and 2-ethyl-7-nitro-5-trifluoromethyl benzimidazole (5).
Each compound identified was identical to authentic standards in chromato-
graphic behavior and mass spectral fragmentation patterns.  It is interesting
to note that methane chemical ionization mass spectrometry (CI/MS) gave
rise to four distinctive fragments.  These were the molecular ion minus 19
(m-19) due to a loss of hydrogen fluoride from the protonated parent molecule
and the m+1, m+9, and Ijl+41 fragments.  The last three components result
from the addition of H , C2H5  to the molecular ion and are characteristic
of methane CI/MS.  All of the authentic standards received from Eli Lilly
and Co. were subjected to GC/CI/MS and each gave rise to these four charac-
teristic fragments.

     In addition to the four metabolites described above, two other com-
ponents designated unknowns A and B were also detected (Table 27).  The GLC
retention time and mass spectrum data for unknown A is indistinguishable
from mono-dealkylated, 2.  Thin-layer chromatography, however, revealed
that product A possessed" a relative R£ of 0.56 while compound 2 has a
relative R^ of 0.82.  This material could be some derivative oF 2 which
yields 2 wnen subjected to elevated temperatures in the GC or GC7MS.  The
other unknown component, B, had a relative R,. of approximately 0.1 to 0.2,
a GC retention time of 9.75 minutes and a mass spectrum different, but
typical of the other dinitro aniline derivatives.  This material has an
apparent molecular weight of 223 and had the m-19, m+29 and m+41 fragments.
A tentative structure has not been identified.

     No differences in metabolic rates or pathways could be detected be-
tween the Webster or Cecil soils or between the formulated or technical
trifluralin.   The quantity of "bound" residues of trifluralin is illustrated
in Figure 27.  The percentage bound was calculated by measuring the organic
solvent extractable 14Carbon and the unextractable amount as determined by
combustion of the soil after extraction.  For the 10 ppm concentration, a
greater percentage of technical trifluralin was bound to Webster soil than
to Cecil.  Ten days after application, 10% was bound and by 35 days it had
risen to 30%  and after 63 days of incubation, 72% was unextractable.

                                      92

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l/J
    TABLE 25.   PERCENTAGE OF APPLIED TRIFLURALIN MINERALIZED AND VOLATILIZED AFTER 83 DAYS OF INCUBATION



                         	10 ppm	     	1,000 ppm	

                                                          On                                       On
                 Soil       Mineralized   Volatilized   Tygon        Mineralized    Volatilized   Tygon
                                                        Tubing                                    Tubing

Webster


Cecil

Technical

Formulated
Technical

Formulated
2.5* (0.04)**

3.1 (0.05)
2.0 (0.03)

1.4 (0.02)
0.01 (.

0.0 (0.
0.0 (0.

0.0 (0.
0015)

0)
0)

0)
7.9

9.8
8.2

8.7
(.12)

(0.4)
(0.12)

(0.13)
0.4

0.5
0.3

0.3
(0.4)

(0.5)
(0.3)

(0.3)
0.0 (0.0)

0.0 (0.0)
0.0 (0.0)

0.0 (0.0)
1.1

1.2
1.2

1.3
(1.1)

(1.2)
(1.2)

(1.3)

     *percent

     **milligrams
                                                                              (continued)

-------
TABLE 25.  (continued)
*percent

**milligrams
                    Soil
                                                   20,000 ppm
                               On
Mineralized   Volatilized    Tygon
                             Tubing
Technical
 Webster
Formulated
Technical
Cecil
Formulated
0.1 (2.0)
0.1 (2.0)
0.1 (2.0)
0.1 (2.0)
0.0 (0.0)
0.0 (0.0)
0.0 (0.0)
0.0 (0.0)
0.1 (2.0)
0.1 (2.0)
0.1 (2.0)
0.1 (2.0)


-------
       TABLE 26.  R£ VALUES  FOR TRIFLURALIN AND METABOLITE REFERENCES



Number                                                                 R


  1     Trifluralin  Ca,a,a-Tri£luoro-2,6-dinitro-N,
        N-dipropyl-p-toluidine                                       1.00

  2     a,a,a-Trifluoro-2,6-dinitro-N-propyl-p-toluidine              0.82

  3     a,a,a-Trifluoro-2,6-dinitro-p-toluidine                       0.25

  4     2-Ethyl-7-nitro-l-propyl-5-tri£luoromethyl benzimidazole      0.25

  5     2-Ethyl-7-nitro-5-tri£luoromethyl  bezimidazole                0

  6     a,a,a-Trifluoro-N,N-dipropyl-5-nitrotoluene-3,4-diamine       0.60

  7     a,a,a-Trifluoro-5-nitro-N-propyltoluene-3,4-diamine           0.22

  8     a,a,a-Trifluoro-5-nitrotoluene-3,4-diamine                    0

  9     a,a,a-Trifluoro-N,N,dipropyltoluene-3,4,5-triamine            0.44

  10     a,a,a-Trifluoromethyl-5-nitrotoluene-3,4-diamine

  11     a,a,a-Trifluorotoluene-3,4,5-triamine                         0
                                      95

-------
TABLE 27.  CHRCMATOGRAPHIC AND MASS SPECTRAL PARAMETERS OF PRODUCTS ISOLATED FROM TRIFLURALIN TREATED
           SOIL

Compound
Trif luralin (a ,a ,a- trif luoro- 2 , 6-
dini tr o -N , N- dipropyl -p - toluidine)
a,a,a-trifluoro-2,6-dinitro-N-
propyl -p - toluidine
Unknown A
a,a,a-trifluoro-2,6-dinitro-
p- toluidine
2 - ethyl - 7 -nitro - 1 -propyl - 5 -
trifluoromethyl benzimidazole
2 - ethyl - 7 -nitro - 5 -
trifluoromethyl benzimidazole
Unknown B
TLCa
Code R,,
1 1.00
2 0.82
0.56
3 0.25
4 0.25
0.00
0.1-0.2
GLCb
Rt
19.6
24.6
24.6
13.2
30.1
22.7
9.75
M-19
316
275
275
234
281
240
204
Massc
M+l
336
295
295
254
301
260
224
Spec
M+29
364
323
323
282
329
288
252
M+41
376
335
335
294
341
300
264

aSee text for TLC parameters; values given are relative Rf's.
 See text for GLC parameters.
GThe four characteristic molecular ions are given.  See text for mass spec parameters.

-------
The quantity of  formulated trifluralin bound in the Webster soil was only
^S^f ?; ??ofd 84  Jay? jncubation>  ^t showed a similar but somewhat
lesser  (45 and 51<)  amount of bound residue than that measured for the
technical material.

     There was less  bound trifluralin material in the Cecil soil.  Approx-
imately 8< of the  technical trifluralin was bound after 7  days.  The per-
centage remained relatively constant throughout the sampling period.  The
formulated material  was somewhat slower in binding, but reached approxi-
mately the same  level.

     For the 1,000 ppm concentration, the  Webster soil also bound more tri-
fluralin than did  the Cecil soil and the technical material appeared to be
bound to a greater extent than did the formulated material.  Twenty to 351
of the  14C was bound in the Webster soil at 1,000 ppm.

     At the 20,000 ppm level, relatively low percentages of trifluralin
were bound; however, these still represent considerable quantities of the
pesticide.  Again  it appears that the Webster soil binds more than Cecil
soil.

     Recent reports  by Katan et al.  (1976)  and Katan and Lichtenstein
(1977) show rapid  binding of the parathion amine  analogue which suggests
that a similar phenomenon may have occurred with  trifluralin in the Webster
soil. The parent compound represented 90%  or more of the extractable lkC
while considerable radioactivity was not extracted.   Thus,  it is possible
that some metabolic  products were formed and a substantial  portion of them
were bound.  This  would explain why certain previously reported metabolites
were not detected  in the  extract or were only found in small quantities.
In an effort to  examine this possibility,  trifluralin,  mono-dealkylated,
and di-dealkylated derivatives were incubated in  sterilized Webster soil
for four hours.  Percentages of each compound extractable  after four hours
were 89, 75 and  57 for the above compounds,  respectively.   Thus, in the
Webster soil there was a  clear relationship between the amount of non-
extractable material and  the substitution  on the  amino nitrogen.  This is
consistent with  the  findings of Katan and  Lichtenstein (1977) with amino
analogs of parathion.   If one fortifies Webster soil with either trifluralin
or the di-dealkylated derivative and immediately  perform an extraction, 93-
95% of both compounds were recoverable.  Substantial binding did not occur
for the di-dealkylated derivative in the sandy Cecil soil.   Thus, it is
possible in the  case of the Webster soil,  that metabolites  containing
secondary or primary amino functional groups became a part  of the "bound"
portion of the residue.   The absence of the same  magnitude  of binding in
Cecil soil could be  the result of different kinds and numbers of microbial
populations, etc.  This could also influence the  formation  of microbially
induced amino metabolites in the Cecil soil.

     The metabolites detected were isolated from  both soils treated at all
three application  rates and are characteristic of the aerobic degradation
pathway described  by Probst et al. (1975)  and shown in Figure 28.  Tri-
fluralin 1 underwent dealkylation to mono-dealdylated and  subsequently


                                       97

-------
80


60


40


20
           o
            o
           o

           u
80


60


40


20
           u
          r
          •«•

           Q


           O
           CQ
80


60


4O


20



 0
                   TRIFLURALIN
                   10 ppm
                   1OOO ppm
20jOOO ppm
WEBSTER TECH. •

WEBSTER FORM.-

CECIL   TECH.

CECIL   FORM.
                      20    40    60    80

                  INCUBATION  TIME (days)
Figure 27.  Percent of total ^C-activity bound to Webster and Cecil soil
          after receiving 10, 1,000 and 20,000 ppm of technical grade
          and formulated trifluralin.
                               98

-------
to the di-dealkylated product.   Two benzimidazoles,  £ (2-ethyl-7-nitro-l-
propyl-5-trifluoromethyl benzimidazole)  and 5_ (2-ethyl-7-nitro-5-triflouro-
methyl bezimidazole)  were also  identified.   Thus, very high application
rates of trifluralin  resulted in the same degradation pathways  that have
been previously reported.

     The GC/MS data system also performed limited mass searches (IMS) for
ions characteristic of 6^ (a,a,a-trifluoro-methyl-N,N-dipropyl-5-nitrotoluene-
3,4-deamine), 7_ (a,a,a-trifluoro-5-nitro-N-propyltoluene-3,4-diamine) and 9^
(a,a,a-Trifluoro-N,N-dipropyltoluene-3,4,5-triamine) in soil extracts from
both soils  (Figure 28).  All LMS's were negative for these compounds. These
materials represent the transitions between postulated aerobic  and anaerobic
pathway.  Thus,  there was no detectable reduction of ring nitro groups.  In
the Webster soil, the possibility exists that the amino metabolites, 6_, 7,
8  (a,a,a-trifluoro-5-nitro-toluene-3,4-diamine)  and 9, may have been hidcfen
In the  "bound" portion of the residue and should notTie overlooked.

2,4-D

     In the majority  of cases, parent 2,4-D was the only radioactive com-
ponent  isolated from the soil.   In a few instances,  2,4-dichlorophenol
 (based  upon thin-layer chromatography) .was detected.  It appeared that more
2,4-dichlorophenol accumulated at the 500 ppm treatment after 7 to 10 days
of incubation than for other treatments and was present to a greater extent
in the  Webster than in the Cecil soil.  Only the parent 2,4-D,  however, was
detected after 29 days of incubation.
                                        99

-------
                           4 ^T       1 '
                              02N
5^V    2
  CF.    .  OF,
                                   \      / CF3\

                                     \H7C3>NH f
                                                   rNH,
Figure 28.  Metabolic pathways for trifluralin metabolism:  Code numbers
            correspond with those in Tables  26 and 27.
                                    100

-------
                                   SECTION 6

     IMPLICATIONS  OF PROJECT RESULTS WITH REGARDS TO PESTICIDE DISPOSAL


     Pesticide  applications to control pests associated with  agricultural,
industrial and  urban environments, in general,  have not had an adverse
effect on soil  microorganisms or groundwater quality.   Because applications
to these environments have involved primarily low concentrations  (0.5 to 10
kg/ha), the  suitability of these data for designing and managing pesticide
waste disposal  sites has been questioned.   Therefore,  the  objectives of
this project were  to:  1)  Measure the adsorption-desorption and mobility
characteristics of various pesticides in different soil-water systems
receiving high  pesticide concentrations; 2) Quantify and describe microbial
degradation  rates  of various pesticides in soils containing large pesticide
concentrations, and 3)  Measure the influence of large  pesticide concentra-
tions on soil microbial activity and respiration rates as  well as to identify
specific microorganisms which degrade a given pesticide at both low and
high pesticide  concentrations.

     Two significant conclusions can be made regarding the adsorption re-
sults obtained  during this project.   First, the Freundlich adsorption equa-
tion described  all pesticide adsorption isotherms considered  for solution
concentrations  up  to the aqueous solubility of  the pesticide.  Thus, the
pesticide adsorption sites for all soils investigated  were apparently not
saturated at any concentration considered in this study.   Second, contrary
to a frequent assumption,  pesticide adsorption  isotherms were not linear,
that is, N in the  Freundlich equation was generally less than one.  The
nonlinearity of the pesticide adsorption isotherm is an important observa-
tion because it explicitly points out that pesticides  will be more mobile
in soils containing pesticide concentrations similar to those associated
with pesticide  waste disposal sites.   This is especially true for pesti-
cides that are  very soluble in water (e.g., 2,4-D amine).

     The increased pesticide mobility at high pesticide concentrations
limits the usefulness of the available low concentration data base for
developing "safe"  management practices for pesticide disposal procedures in
soils.  If a linear adsorption isotherm is assumed on  the  basis of the low
pesticide concentration data, one underestimates the soil  depth to which a
pesticide will  leach or move for a given water  input.   The seriousness of
the failure  of  the low concentration data base  to describe the true mobility
of a pesticide  as  it moves toward the groundwater from a waste disposal
site depends upon  the water solubility of  the pesticide and nonlinearity of
the adsorption  isotherm.   For example, the adsorption  isotherms for atrazme
and 2,4-D and a Eustis  soil were similar (see Table 5); however, 2,4-D


                                      101

-------
moved similar to an unadsorbed chemical as it moved away from a simulated
waste disposal site, while the mobility of the atrazine through the same
soil was about 2.5 times less (Figure 14).  To have assumed that the adsorp-
tion isotherms were linear would have resulted in a serious underestimation
of the depth to which each pesticide would have moved for a given water
input.

     Many of the pesticides available on the market today are biodegradable
in soils when applied at low concentrations (0.5 to 10 kg/ha).  However,
many of these same organic chemicals become persistant when applied to
soils at high concentrations.  It has been observed that some soils are
able to biologically mineralize one pesticide (e.g., 2,4-D in Webster
soil), but the same pesticide may be persistant in another soil (e.g.,
Cecil). This study clearly points out that the soil respiration rate of a
soil receiving high pesticide concentrations is not, in general, a reasonable
procedure for measuring pesticide degradation potential.  Also, the apparent
persistance of some pesticides may be further confounded by the formation
of metabolites which are "bound" (not extractable by recommended procedures)
to the soil and suggest a greater apparent loss of the original chemical
than what actually occurred.

     The contrast between the behavior of soil environments containing
large and small pesticide concentrations illustrates the importance and
potential for management of pesticide waste disposal sites.  Many soils
frequently can be altered chemically and/or biologically to enhance their
potential for biologically degrading pesticides.  Also, microorganisms
capable of degrading specific chemicals at high concentrations have been
identified and isolated which could be added to a waste disposal site to
enhance the degradation and mineralization of a given chemical.  Because of
increased pesticide mobility at high concentrations, the chemical may not,
however, remain in the vicinity of desired biological environment for
degradation; thus, it is important to manage both the water leaching rate
and biological environment for optimum inactivation and efficiency of a
pesticide waste disposal site.

     A major limitation in using the results of this project is that only
one chemical was applied to a soil at a time.  Combination or mixtures of
pesticides would be the common situation for a pesticide waste disposal
site.  Waste disposal sites receiving several pesticides may fail to func-
tion as designed for a specific pesticide because of interactions between
chemicals and their environment.  The behavior of a pesticide mixture may
or may not be independent or additive, but rather based upon the influence
of one pesticide and/or the formulation associated with a given pesticide.
Problems which may arise owing to the mixing of pesticide in a waste disposal
site should be considered before site selection and management protocols
are defined by the United States Environmental Protection Agency.  This
work should include the evaluation of the major surfactants used with
pesticides and various formulation chemicals.
                                     102

-------
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Atkins, P. R.   The Pesticide Manufacturing Indus try--Current Waste Treat-
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Bailey, G. W.  and J.  L.  White.   Factors Influencing Adsorption, Desorption,
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Breazeale, F.  W., and N. D.  Camper.  Bacterial, Fungal, and Actinomycete
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Cameron, D. A.  and A.  Klute.  Convective-Dispersive Solute Transport
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Cole, M. A.  Effect of Long-Term Atrazine Application on Soil Microbial
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Dao, T. H. and T. L.  Lavy.   Factors Affecting Atrazine Adsorption on Soil
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Davidson, J. M.,  L.  T. Ou,  and  P. S.  C.  Rao.  Behavior  of High Pesticide
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Davidson, J. M. and R. K. Chang.   Transport  of  Picloram in Relation to
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-------
Eno, C. F.  The Effect of Simazine and Atrazine on Certain of the Soil
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Foster, R. K., and R. B. McKercher.  Laboratory Incubation Studies of
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Hamaker, J. W.  The Interpretation of Soil Leaching Experiments.  In;
     Environmental Dynamics of Pesticides (eds. R. Haque and V.  H."Freed).
     Plenum Press, N. Y.  1975.  p. 115-131.

Hamaker, J. W. and J. M. Thompson.  Adsorption.  In:Organic Chemicals
     in the Soil Environment,  (eds. c. A. I. Goring and J. W. Hamaker).
     Vol. 1.  Marcel Dekker, Inc., N. Y.  1972.  p. 49-143.

Hance, R. J.  The Adsorption of Atrazine' and Monuron by Soils at Different
     Water Contents.  Weed Res. 17:197-201, 1977.

Hornsby, A. G. and J. M. Davidson.  Solution and Adsorbed Fluometuron
     Concentration Distributions in a Water-Saturated Soil:  Experimental
     and Predicted Evaluation.  Soil Sci. Soc. Amer.  Proc. 37:823-828,
     1973.

Hubbell, D. H., D. F. Rothwell, W. B. Wheeler, W. B.  Tappan, and F. M.
     Rhoads.  Microbiological Effects and Persistence of Some Pesticide
     Combinations in Soil.  J. Environ. Quality 2:96-99, 1973.

Hugenberger , F., J. Letey and W. J. Farmer.  Observed and Calculated
     Distribution of Lindane in Soil Columns as Influenced by Water Move-
     ment.  Soil Sci. Soc. Amer. Proc. 36:544-548,  1972.

Kaiser, P., J. J. Pochon, and R. Cassini.  Influence of Triazine Herbi-
     cides on Soil Microorganisms.  Residues Rev. 32:211-233,  1970.

                                     104

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Katan, J., T. W. Fuhreman,  and E.  P.  Lichtenstein.  Binding of
     Parathion in Soil:  A  reassessment  of Pesticide Persistence
     Science 195, 891-894,   1976.

Katan, J., and E. P. Lichtenstein.  Mechanisms  of Production of Soil-
     Bound Residues of  [lltC]  Parathion by Microorganisms.  J. Aer
     Food Chem. 25, 1404-1408,  1977.

Kay, B. D. and D. E. Elrick.   Adsorption and Movement of Lindane in Soils.
     Soil Sci. 104, 314-322,   1967.

Kirkham,  D. and W. L. Powers.  Advanced  Soil Physics.  Wiley-Inter-
     science, New York.  1972.  238 pp.

Lambert,  S. M.  Omega  (n),  A Useful Index of Soil Sorption Equilibria.
     J. Agr. Food. Chem. 16:340-343,   1968.

Lichtenstein, E. P., J. Katan, and B.  N.  Anderegg.  Binding of "Persistent"
     and  "Nonpersistent"  ltfC-Labeled  Insecticides in an Agricultural Soil.
     J. Agr. Food Chem. 25:43-47,  1977.

Lindsey,  A. W., D. Farb and W. Sanjour.   OSWMP  Chemical Waste Landfill and
     Related Projects.  In:Residue Management by Land Disposal:  Proceed-
     ings of the Hazardous  Waste Research Symp.  (ed. W. H. Fuller).  EPA-
     600/9-76-015, U. S. Environmental Protection Agency, Cincinnati,
     Ohio, 1976.  p. 14-31.

Loos, M.  A.  Phenoxylalkanoic Acid.   In:Herbicides, Vol. 1 (eds. P. C.
     Kearney and D. D.  Kaufman).   Dekk~er, New York.  1975.  p. 1-128.

Loos, M.  A.  Phenoxylalkanoic Acids.   In:Degradation of Herbicides (eds.
     P; C. Kearney and  D. D.  Kaufman).   Dekker,  New York.  1969.  p. 1-49.

Martin, J. P.  Use of Acid, Rose Bengal  and Streptomycin in Plate Method
     for  Estimating Soil Fungi.  Soil Sci.  69:215-222,  1950.

Marquardt, D. L.  An Algorithm for Least-Squares Estimation of Non-
     Linear Parameters.  J.  Soc. Indust.  Appl. Math. 2:431-441,  1963.

Meeter, D. A. and P. J. Wolfe.  UWHAUS-Nonlinear Least-Squares Fitting
     and  Function Minimization.  Univ. of Wisconsin Computing Center,
     Madison, Wisconsin.  1968.

Nearpass, D. C.  Effect of  Predominating Cation on the Adsorption of
     Atrazine by Bayboro Clay Soil.   Soil Sci.  103:177-182,  1967.

Newman, A. S. and C. R. Downing.   What Pesticides do to Soil.  3. Herbi-
     cides and the Soil.  J.  Agr.  Food Chem. 6:342-353.  1958.

O'Connor, G. A., M. Th. van Genuchten, and P. J. Wierenga.  Predicting
     2,4,5-T Movement in Soil Columns.   J. Environ. Qual. 5:375-378,
     1976.

                                     105

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Ou, L. T., J. M. Davidson and D. F. Rothwell.  Response of Soil Microflora
     to High 2,4-D Concentrations on Degradation and Carbon Dioxide Evolu-
     tion in Soils.  J. Environ. Quality 7:241-246,  1978a.

Ou, L. T., J. M. Davidson and D. F. Rothwell.  Response of Soil Microflora
     to High 2,4-D Applications.  Soil Bio. Biochem. 10;443-445,  1978b.

Probst, G. W., T. Golab, and W. L. Wright.  Dinitroamines.  In:P. C.
     Kearney and D. D. Kaufman (eds.) Herbicides; Vol. 1.  Marcel Dekker,
     New York.  1975.

Rao, P. S. C., J. M. Davidson, R. E. Jessup, and H. M. Selim.  Evaluation
     of Conceptual Models for Describing Nonequilibrium Adsorption-
     Desorption of Pesticides During Steady-Flow in Soils.  Soil Sci.
     Soc. Amer. J. Vol. 43(1) (in press),  1979.

Rao, P. S. C., R. E. Green, L. R. Ahuja, and J. M. Davidson.  Evaluation
     of a Capillary Bundle Model for Describing Solute Dispersion in
     Aggregated Soils.  Soil Sci. Soc. Amer. J. 40:815-820,  1976.

Rose, D. A. and J. B. Passioura.  The Analysis of Experiments on Hydro-
     dynamic Dispersion.  Soil Sci. 111:252-257,  1972.

Roslycky, E. B.  Response of Soil Microbiota to Selected Herbicide Treat-
     ments.  Can. J. Microbiol. 23:426-433,  1977.

Rouston, R. C. and R. B. Wildung.  Ultimate Disposal of Wastes to Soil.
     In:Water  (ed. L. K. Cecil).  Chem. Eng. Progress Symp.  Series
     64.97, 1969.  p. 19-25.

Sanborn, J. R., B. F. Francis, and R L. Metcalf.  The Degradation of
     Selected Pesticides in Soil:  A review of published literature.
     U. S. Environmental Protection Agency, Cincinnati, Ohio, 1977.
      (EPA-600/9-77-022, August 1977).

Schomaker, N. B.  Current Research on Land Disposal of Hazardous Wastes.
     In:Residue Management by Land Disposal:  Proc. of the Hazardous Waste
     Res. Symp. (ed. W. H. Fuller).  EPA-500/9-76-015, U. S. Environmental
     Protection Agency, Cincinnati, Ohio, 1976.  p. 1-13.

Selim, H. M., J. M. Davidson, and R. S. Mansell.  Evaluation of a Two-
     Site Adsorption-Desorption Model for Describing Solute Transport
     in Soils.  Proc. of Summer Computer Simulation Conf. Washington,
     D. C., p. 444-448.  1976.

Skopp, J. and A. W. Warrick.  A Two-Phase Model for the Miscible Dis-
     placement of Reactive Solutes Through Soils.  Soil Sci. Soc. Amer.
     Proc. 38:545-550, 1974.

Skopp, J., E. J. Tyler and W. R. Gardner.  An Interacting Two-Pore Model
     for Solute Dispersion in Aggregated Soils.  Agron. Abst.  1977.
     p. 53.

                                     106

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Stojanovic, B. J., M. V. Kennedy,  and F.  L.  Shuman, Jr.  Edaphic Aspects
     of the Disposal of Unused Pesticides, Pesticide Wastes, and Pesticide
     Containers.  J. Environ.  Quality 1:54-62,  1972.

Tyunaeva, G. N., A. K. Minenko, and L.  A. Pen'kov.  Effect of Treflan on
     the Biological Properties of  the Soil.  Agrokhimiya 1974:110-114,
     xy / *f •

Valoras, N., J. Letey, and F.  J. Osborn.  Adsorption of Non-Ionic Sur-
     factants by Soil Minerals. Soil Sci. Soc. Amer. Proc. 33:345-348
     1969.                                                            '

Van Genuchten, M. Th., and P.  J. Wierenga.   Mass Transfer Studies in
     Sorbing Porous Media.   II. Experimental Evaluation with Tritium
     (H20).  Soil Sci. Soc.  Amer.  J.  41:272-278,  1977.

Van Genuchten, M. Th., and P.  J. Wierenga.   Mass Transfer Studies in
     Sorbing Porous Media:   I.  Analytical Solutions.  Soil Sci. Soc.
     Amer. J. 40:473-480,   1976.

Van Genuchten, M. Th., J.  M. Davidson and P. J. Wierenga.  An Evaluation
     of Kinetic and Equilibrium Equations for Predicting Pesticide Move-
     ment Through Porous Media. Soil Sci. Soc. Amer. Proc. 38:29-35,
     1974.

Voets, J. P., P. Meerschman, and W.  Verstraete.  Soil Microbiological
     and Biochemical Effects of Long-Term Atrazine Applications.  Soil
     Biol. Biochem. 6:149-152,  1974.

Von Everdingen, R. 0. and  R. A. Freeze.   Subsurface Disposal of Waste
     in Canada.  Tech. Bull. NO. 49.   Inland Waters Branch, Dept. of Envi-
     ronment, Ottawa, Canada,  1971.   19 pp.

Von Rumker, R., E. W. Lawless,  A.  F.  Meiners, K. A. Lawrence, G. L.
     Kelso, and F. Horay.   Production,  Distribution, Use and Environmental
     Impact Potential of Selected  Pesticides.  EPA-540/1-74-001, U. S.
     Environmental Protection  Agency, Cincinnati, Ohio, 1974.  439 pp.

Watts, R. R.  Extraction Efficiency Study--Examination of Three Procedures
     for Extracting C-Labeled  and  Uhlabeled  Residues of Organophosphorus
     Pesticide and Carbaryl from Bean Leaves and Kale.  Jour, of the
     Association of Official Analytical Chemists 54:953-958, 1971.

Weber, W. J. and P. J. Usinowicz.  Adsorption from Aqueous Solution.
     Tech. Publication, Research Project  17020 EPA, U. S. Environ. Prot.
     Agency, Cincinnati, Ohio.  1973.  187 pp.

Wilkinson, R. R., G. L. Kelso,  and F. C. Hopkins.  State-of-the-Art Report:
     Pesticide Disposal Research.   EPA (in press) U. S. Environmental
     Protection Agency, Cincinnati,  Ohio, 1978.  237 pp.
                                      107

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Williams, S. T. and F. L. Davis.  Use of Anitbiotics for Selective Iso-
     lation and Enumeration of Actinomycetes in Soil.  J. Gen. Microbiol.,
     38:251-261, 1965.

Wolfe, H. R., D. C. Staff, J. F. Amstrong, and S. W. Coiner.  Persistence
     of Parathion in Soil.  Bull. Environ. Contamin. Toxicol. 10:1-9,
     1973.

Wood, A. L. and J. M. Davidson.  Fluometuron and Water Content Distribu-
     tions During Infiltration:  Measured and Calculated.  Soil Sci. Soc.
     Amer. Proc. 39:820-825.  1975.
                                     108

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                                 APPENDIX

             LIST OF PUBLICATIONS  RESULTING FROM THIS PROJECT


1.  Davidson, J. M., L.-T. Ou,  and P.  S.  C. Rao.  1976..  Behavior of high
    pesticide concentrations  in soil-water systems.  In:Residual Manage-
    ment by Land Disposal:  Proc.  of the  Hazardous Waste Research Symp.
    (ed. W. H. Fuller).   EPA-600/9-76-015, July  1976.  p. 206-212.

2.  Rao, P. S. C., J. M.  Davidson, and L. C. Hammond.  1976.  Estimation
    of nonreactive and reactive solute front locations in soils.  In:
    Residual Management Waste Research Symp. (ed. W. H. Fuller).  EPA-
    600/9-76-015, July 1976.  p. 235-242.

3.  Ou, L.-T., D. F. Rothwell,  W.  B. Wheeler, and J. M. Davidson.  1978.
    The effect of high 2,4-D  concentrations on degradation and carbon
    dioxide evolution in  soils.  J.  Environ. Quality.  7_, 241-246.

4.  Davidson, J. M., L.-T. Ou,  and P.  S.  C. Rao.  1978.  Adsorption,
    movement, and biological  degradation  of high concentrations of
    selected pesticide in soils.   In:Fourth Annual Hazardous Waste
    Management Symp. (ed. D.  ShultzJ.   EPA-600/9-78-016, March 1978.

5.  Ou, L.-T., J. M. Davidson,  and D.  F.  Rothwell.  1978.  Response of
    soil microflora to high 2,4-D  applications.  Soil Biol. Biochem.
    10;443-445, 1978.

6.  Rao, P. S. C., and J. M.  Davidson.  1978.  Adsorption and movement
    of selected pesticides at high concentrations in soils.  Water Res.
    (in press).

7.  Rao, P. S. C., J. M.  Davidson, R.  E.  Jessup, and H. M. Selim.  1978.
    Evaluation of conceptual  models for describing nonequilibrium adsorp-
    tion-desorption of pesticides  during  steady-flow in soils.  Soil
    Sci. Soc. Amer. J. Vol. 43(1), 1979.

8.  Ou, L.-T., and J. M.  Davidson.  High  concentration effect of methyl-
    parathion on degradation  and carbon dioxide  evolution in soils.  J.
    Environ. Quality,  (submitted).

9.  Ou, L.-T., J. M. Davidson,  and D.  F.  Rothwell.  High concentration
    effect of herbicides  trifluralin and  atrazine on microbiota and soil
    respiration.  Soil Biol.  Biochem.   (submitted).
                                    109

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10.   Wheeler, W. B.,  G.  D.  Stratton,  R.  P.  Twilley, L.-T.  Ou, D.  A.  Carlson,
     and J. M. Davidson.   1979.   Trifluralin degradation and binding in
     soils at high concentrations.  J. Agr.  Food Chem.   (submitted).

11.   Rao, P. S. C. and J. M.  Davidson.   Non-equilibrium conditions for
     solute transport in soils:   Flow interuption experiments.  Soil Sci.
     (in preparation).

12.   Wheeler, W. B.,  G.  D.  Stratton,  R.  P.  Twilley, L.-T.  Ou, and J. M.
     Davidson.  Atrazine degradation  and binding in soils  at high appli-
     cations .  (in preparation).
                                    110

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                                   TECHNICAL REPORT DATA
                            (flease read Instructions on the reverse before completing)
 EPA-600/2-80-124
                                                           3. RECIPIENT'S ACCESSION-NO.
          iUBI
              ri_E
Adsorption,  Movement, and  Biological Degradation  of
Large  Concentrations of  Selected Pesticides  in  Soils
                                     5. REPORT DATE

                                       August 1980 (Issuing Date)
                                     6. PERFORMING ORGANIZATION CODE
 P.  S.  C.  Rao
L. T. Ou
W. B. Wheeler
                                         D. F. Rothwell
                                                           8. PERFORMING ORGANIZATION REPORT NO
                 IZATION NAME AND ADDRESS
 Soil  Science Department
 University of Florida
 Gainesville, Florida
                                     10. PROGRAM ELEMENT NO.

                                       1DC618
                                     11. CONTRACT/GRANT NO.

                                       R-803849
 .12.SP-QNSORING AGENCY NAME AND ADDRESS
 Municipal  Environmental  Research Laboratory—Cin., OH
 Office  of Research and  Development
 U.S.  Environmental Protection Agency
 Cincinnati,  Ohio 45268
                                     13. TYPE OF REPORT AND PERIOD COVERED
                                       8/1/75 to  10/31/77
                                     14. SPONSORING AGENCY CODE


                                       EPA/600/14
 15. SUPPLEMENTARY NOTES
 Project Officer:  Mike  H.  Roulier  (513) 684-7871
 is.ABSTRACT because or-tne importance ot soil  in  biologically reducing the quantity and
 retarding the rate  of pollutant movement  into groundwater,  this laboratory study was
 initiated to evaluate the adsorption, mobility,  and  degradation of large concentrations
 of the pesticides atrazine, methyl parathion, terbacil, trifluralinland 2, 4-D in soils
 representing four major soil  orders in the  United  States.
      Solution concentrations ranged from  zero to the aqueous solubility limit for each
 pesticide.   The mobility of each pesticide  increased as its concentration in the soil
 solution phase increased.   These results  were in agreement  with the adsorption isothern
 data.   Pesticide degradation rates and soil microbial  populations generally declined
 as the pesticide concentration in soil increased;  however,  some soils were able to
 degrade a pesticide at all  concentrations studied, while others remained essentially
 sterile throughout  the incubation period  (50 to  80 days).   As shown by measurements of
 ltfC02  evolution, total  C02  evolution was  not always  a  good  indication of pesticide
 degradation.  Several  pesticide metabolites were formed and identified.  Bound residues
 of trifluralin and  atrazine at the end of the incubation period appeared to be related
 to types of metabolites formed.
      The observed increase  in pesticide mobility for large  pesticide concentrations in
 the soil invalidates, .in many cases, the  usefuln^o  of the  existing low concentration
 data base for designing pesticide waste disposal sites.
17.
                                KEY WORDS AND DOCUMENT ANALYSIS
                  DESCRIPTORS
                                              b.lDENTIFIERS/OPEN ENDED TERMS
                                                  c. COSATI Field/Group
      Pesticides
      Soil  Chemistry
      Adsorption
      Soils
      Mobility
      Degradation
                         Pollutant Migration
                         Groundwater Pollution
13B
 8. DISTRIBUTION STATEMENT
      Release to Public
                        19. SECURITY CLASS (ThisReport)'
                          Unclassified
                                                                         21. NO. OF PAGES
                                                                           123
                        20. SECURITY CLASS (Thispage)
                          Unclassified
                                                                        22. PRICE
EPA Form 2220-1 (9-73)
                                            111
                                             4 U.S. GOVERNMENT PRINTING OFFICE: 1980-657-165/0029

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