United States
         ' Environmental Protection
          Agency
             Office of Water
             4304
EPA822-D-Q1-002
August 2001
SEPA
Ambient Aquatic Life
Water Quality Criteria
for Atrazine - Draft

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AMBIENT AQUATIC LIFE WATER QUALITY CRITERIA FOR

                   . .ATRAZINE

           CAS Registry No. 1912-24-9
     U.S. ENVIRONMENTAL PROTECTION AGENCY

                OFFICE OF WATER
       OFFICE OF  SCIENCE AND TECHNOLOGY
    HEALTH AND ECOLOGICAL CRITERIA DIVISION
                WASHINGTON D.C.

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                                   NOTICES
This document has been reviewed by the Health and Ecological Criteria
Division, Office of Science and Technology, U.S. Environmental Protection
Agency, and approved for publication.

Mention of trade names or commercial products does not constitute endorsement
or recommendation for use.

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                               EXECUTIVE SUMMARY









 BACKGROUND:









      Atrazine is an herbicide that has  been used extensively throughout  the




 U.S.  for control of weeds in agricultural  crops.   It  is  a  problem in  the




 aquatic environment because it is  toxic to aquatic organisms and  is often




 found in surface water.'  EPA is developing ambient water quality  criteria for




 atrazine through its authority-under  Section 304(a) of the Clean  Water Act




 (CWA).   These water quality criteria  may be used by States and  Tribes to




 establish water quality standards  for atrazine.









 CRITERIA:  .









 Freshwater:




      For atrazine,  the  criterion to protect freshwater' aquatic  life from




 chronic  toxic effects is  a  four-day average of  12  ug/L,  not to  be exceeded




 more  than  once every three  years on the average.   The criterion to protect




 freshwater aquatic  life from  acute toxic effects  is a one-hour .average of 350




 ug/L, not  to  be  exceeded  more  than once every three years  on the  average.








 Saltwater:




      For atrazine,  the criterion to protect saltwater aquatic life from




•chronic -toxic effects is  a  four-day average of  26  ug/L,  not to  be exceeded ,




 more  than  once every three  years on the average.   The criterion to protect




 saltwater  aquatic life from acute toxic effects  is a one-hour average of 760




 ug/L, not  to  be  exceeded  more  than once every three years  .on the  average.
                                     111

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      The criteria for atrazine were developed by the EPA Office  of  Water (OW)




 using a•large aquatic toxicity data base.   Adverse effects  of  atrazine on




 survival,  growth, and reproduction of aquatic organisms  were demonstrated in




 numerous laboratory studies.   In addition,  field studies have  also



 demonstrated the toxicity of  atrazine to aquatic life.








      Concurrent with the publication of these water quality criteria  for



 atrazine,  the EPA Office of Pesticide Programs (OPP)  is  publishing  its




 Preliminary Ecological  Pate and Effects Assessment for Atrazine.  Both EPA




.offices  shared their aquatic  toxicity data  bases for atrazine  in the




 development of their respective ecological  risk  assesssment documents.




 OW  and OPP are currently consulting on their  respective  ecological  risk



 assessment methodologies.  . Although there are similarities  "in  the approaches.,



 differences remain.   While this consultation  is  underway, both EPA  offices are



 making their ecological  risk  assessments for  atrazine available  to .the public



 and requesting comment on  their respective  methodologies.








     This  document-provides guidance to'States and Tribes authorized  to



 establish  water quality  standards under the Clean  Water Act (CWA) to  protect



 aquatic life  from acute  and chronic  effects of atrazine.  Under  the CWA,



 States and  Tribes are to establish water quality criteria to protect



 designated  uses.  While this document constitutes U.S.EPA's  scientific



 recommendations regarding  ambient concentrations of atrazine,  this  document



 does not substitute  for  the CWA or U.S.EPA's  regulations; nor  is  a  regulation



 itself.  Thus, it cannot impose legally binding  requirements on U.S.EPA,



 States, Tribes, or the regulated community, and  it might not apply  to a




particular  situation based upon the  circumstances.   State and  Tribal  decision-



makers retain  the discretion  to adopt  approaches on a case-by-case  basis that




differ from this guidance when  appropriate.   U.S.EPA may change  this  guidance



in the future.

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                                ACKNOWLEDGMENTS
 Daniel J.  Call
 University of Wisconsin-Superior
 Superior,  Wisconsin

 Tyler K. Linton
 Great Lakes Environmental  Center
 Columbus,  OH

 Frank Gostomski
.(document  coordinator)
 U.S.  EPA
 Health and Ecological  Criteria Division
 Washington,  D.C.
                                       IV

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                                    CONTENTS

 NOTICES ................  .


 EXECUTIVE SUMMARY .  .  . ,
                        , ................  ...  ......   iii

 ACKNOWLEDGMENTS ........... ..........  .......  .  .  iv


 TABLES  ........ .............  ........  .....  vi


 FIGURES ........... ....... .........  .......   vii


 Introduction  ....... ..... ........  ........  .  .     i


 Acute  Toxicity to  Aquatic  Animals .  ..............  ......   9


 Chronic Toxicity to Aquatic Animals  ...... .....  .......  .  .  12


 Toxicity to  Aquatic Plants  ..... .  .  .............  ....  19


 Bioaccumulation ...... ....  ....................  22

 Other  Data   .....  ...................  .  .......  23


 Unused Data  .................. ...............  50

 Summary .........  ........... ...........        53
                                          •      _/                   .
National Criteria  .  .  .  .  ........  .  .  . .  . .  .  .  .  .  .    .  .  ,  __  f  57


 Implementation   ' .................. .  ...........  5-7

References   ......  ...... ' .........  ........ .  ^  ^  152
                                       v

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                                    TABLES







                                                                   .     -  Page




1.  Acute Toxicity of Atrazine to Aquatic Animals .... 	 65




2a. Chronic Toxicity of Atrazine to Aquatic Animals '	,68-




2b. Acute-Chronic Ratios	 70




3.  Ranked Genus Mean Acute Values with Species Mean Acute-Chronic Ratios . 71




4.  Toxicity of Atrazine to Aquatic Plants	74




5.  Bioaccumulation of Atrazine by Aquatic Organisms  	 81




6.  Other Data on Effects of Atrazine on Aquatic Organisms	82
                                      va

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                                    FIGURES




                                                                           Page

1.    Ranked  Summary of  Atrazine GMAVs  -  Freshwater 	  60

2.    Ranked  Summary of  Atrazine GMAVs  -  Saltwater  	  61

3.    Chronic Toxicity of  Atrazine  to Aquatic Animals  ...........  62
                                '>
4.    Ranked  Summary of  Test Values for Freshwater Plants	63

5.    Ranked  Summary of  Test Values for Saltwater Plants	64
                                      VI1

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Introduction1



      Atrazine  is  a herbicide with  the  empirical  formula C8H14C1SNS and a

molecular  weight of 215.7.   It  is a white,  crystalline  solid with a melting

point of 173-175°C, a boiling point of 279°C,  and solubility in water of 33

mg/L at 25°C  (Farm  Chemicals Handbook 2000; Hunter et al. 1985) .  Atrazine has
                                u
an n-octanol-water partition coefficient  (log P)  of  2.82,  a vapor pressure of

7.34 x 1Q-* mm Hg,  a Henry's Constant  of 8.32  x 10'6 atarmVM,  and  a  hydrolysis

half-life  in  excess of  1,000 days  (Hunter et  al.  1985).   These physico-

chemical properties contribute  to its environmental  partitioning and degree  of

persistence in  the aquatic  environment.

      Atrazine  is  used  extensively  in the United States, Canada and other

countries  for control of weeds  in agricultural crops,  especially in crops such

as corn, sorghum,  wheat and soybeans.   It is  one of  the most heavily used

pesticides in North America, generally  being  among the top few in terms of

total pounds  of herbicide used  (Braden  et al. 1989;  Burridge and Haya 1988;

Ciba-Geigy 1994; Council on Environmental Quality 1984; Moxley 1989; Pike

1985; Richards  and Baker 1993). Annual domestic usage during the past two

decades has been in the general range of 30 to 40 million kilograms applied to

approximately 70 million acres  of  farm  land in the U.S.  (U.S. EPA  2000).  It

is also commonly used in other  countries (Bester and Huhnerfuss  1993; Bester

et al. 1995;  Caux  and Kent  1995;  Galassi et al. 1992, 1993; Lode et al. 1994).

Atrazine  is also used in combination with other herbicides  including alachlor,
       "•An understanding of the "Guidelines for Deriving Numerical National
Water Quality Criteria for the Protection of Aquatic Organisms  and Their Uses"
 (Stephen et al. 1985) ,. hereafter referred to as the Guidelines,  is necessary
in  order to understand the following text, tables and  calculations.

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 ametryne,  liriuron, paraquat, propachlor,  amitrole,  and cyanazine  (Farm


 Chemicals  Handbook 2000).


       With this magnitude of application,  atrazine .has commonly been detected


 in  surface waters of agricultural watersheds where it has been used.  Due to


 its  relative mobility from soil, atrazine surface water concentrations are


 highest in field runoff, with concentration peaks generally following early

                               >'              - '                •
 major  storm events that occur within a few weeks of application  (Glotfelty et


 al.  1984;  Muir et al.  1978; Triplett et al.  1978; Wauchope 1978; Wauchope and


 Leonard 1980} .  Concentrations in the low mg/L range may be encountered in


 edge-of-field run-off  (Hall et al.  1972;  Kadoum and Mock 1978; Klaine et al.


 1988;  Roberts et al. • 1979).  Field' run-off is diluted upon entering a stream


 or lake, resulting in  atrazine concentrations that are generally much lower


 (e.g.,  1-10  ^g/L range)  in such waters (Frank and Sirons 1979; Frank et al.


 1979;  Richards and Baker 1993; Richard et al. 1975; Roberts et al. 1979; Wu


 1981).  • Only trace levels (i.e., <1.0-33  ng/L)  were reported in a pesticide

                       ,      ,,. ;        , " •.                     ,               •
monitoring study in California  (Pereira et al.  1996).  However, individual


maximum concentrations may be considerably higher.   When considered over


 several  years,  maximum concentrations reported in some creeks and rivers from


midwestern agricultural  areas have  ranged from 5 to 70 ;/g/Ii  (Ciba-Geigy


 1992a,b,c,d,  1994; Frank and Sirons 1979;  Frank et al. 1979, 1982; Illinois


 State  Water  Survey 1990; Muir et al. 1978; Richards and Baker 1993; Roberts et


 al.  1979) .   Factors that strongly and positively correlate with the release of


 atrazine from-soil include sediment organic carbon, landscape position, and


 tillage  (Novak 1999) .


       Surface waters surrounded by agricultural lands may receive several


pulsed doses  over the  growing season corresponding to. rainfall events  (Herman


 et al.  1986) .   Annual  patterns of atrazine. concentrations in Ohio streams  show

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peak time-weighted mean concentrations of about 6 ^g/L in early June, with a

rapid increase from April to June, followed by a rapid decrease from June to

August (Richards and Baker 1993).  Time-weighted mean concentrations between

August and December are considerably lower, most frequently being less than

1.0  A*g/L-   Atrazine concentrations are the lowest,  and uniformly so, between

January and April.  Also, smaller streams were shown to have higher peak
                                11                                •
concentrations, but of shorter duration,  than larger streams (Richards and

Baker 1993) .   The annual cycle is similar in southwestern Ontari.o, but with

the  annual peak concentrations occurring  at lower levels and several weeks

later than in Ohio (Bodo 1991).  Nonetheless,  atrazine concentrations in

Ontario have  regularly exceeded 2 ^g/L, which is the Canadian water quality

guideline  for aquatic life protection (Trotter et al.  1990).  Exceedances have

similarly  been reported in surface waters of Quebec (Caux and Kent 1995).

       Among the highest surface water concentrations of atrazine are those in

small reservoirs in southern Illinois. These are currently being intensively

monitored  (Tierney et al. 1994a) .  Maximum concentrations as high as 55 Mg/L

have  been  reported from these reservoirs.

       Similar seasonal trends in concentrations of atrazine to those in Ohio

streams have  been observed in streams in  Illinois (Ciba-Geigy 1992a; Illinois

State Water Survey 1990), in Iowa (Ciba-Geigy 1994), and in other midwestern

states (Ciba-Geigy 1992c) .  In large rivers such as the Mississippi, Missouri

and Ohio Rivers, peak concentrations have most commonly occurred in. June, with

mean  levels of less than 5.0 ^g/L during  the spring period  (Ciba-Geigy 1992b) .

The maximum concentrations were generally between 2 and 8 A*g/L, with a single

maximum as high as 17.25 ^g/L  (Ciba-Geigy 1992b,c).' Atrazine concentrations

in the Mississippi River between Minneapolis,  Minnesota and New Orleans,

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  Louisiana from July to August, 1991 ranged from 0.054 /^g/L to 4.7 /zg/L

  (Pereira and Hostetler 1993).

        Atrazine residues in Illinois lakes tended to be lower than those in the

  streams (with less pronounced peak values),  however,  the lower concentrations

  were  sustained for longer durations (Ciba-Geigy 1992a) .   It should be noted

  that  the maximum observed atrazine concentration was  less than 3 .0 /zg/L at 61
                                 i|          :                    .
,  percent of 42 sites monitored over 6 years between 1975  and 1988 (Ciba-Geigy

  1992a).


        Atrazine concentrations were .considerably lower in Chesapeake Bay and

  its tributaries (Ciba-Geigy 1992e) .  Here, the maximum observed concentration

  in a  tributary was 14.6 ,ug/L, and only three out .of 600  samples analyzed

 between  1976  and 1991  exceeded 3 .0 Mg/L.   The highest observed maxima in the

 Upper  and  Lower Chesapeake Bay were 1.7  and  0.38 W/L, respectively.  Models

 for the  Great. Lakes suggest that concentrations should be quite low, not '

 likely  to  exceed 0.13  /zg/L (Tierney et al. 1994b) .   Individual measurements

 from Lake  Erie taken at Toledo,  Ohio,  have not exceeded  0.35 ;zg/L,  while

 concentrations measured from samples collected in Lake Michigan at Michigan

 citv'  Indiana,  have been below 0.20 i/g/L  (Ciba-Geigy  1992c) .

       In addition  to field run-off,  atrazine residues are also transported by

 volatilization into the atmosphere and subsequent deposition.  Atrazine has

 been measured in fog (Glotfelty et al.  1987),  and trace  amounts have been

 shown to be transported by the wind (Elling  et al.  1987).  Atrazine was

 present year-round in  rainwater samples  in Maryland,  with the highest

 concentration of 2.2 Azg/L  occurring in May (Wu 1981) .

       Atrazine has  been shown to be enriched at the microsurface layer of

 water  (Wu 1981; Wu et  al.  1980).   This may be due to  the presence of

 microsurface  films  which tend to concentrate certain  chemicals.  Wu (1981)

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suggested  that atrazine enrichment  in  the microsurface layer was more likely a

source  of  direct input rather  than  a result of  atmospheric wet  deposition, and

that  the main source of atrazine  at the  site  studied  in Maryland was

agricultural runoff.

      Studies of atrazine  persistence  in water  have produced varying results.

Huckins et al. (1986)  reported the  loss  of atrazine from water  within 4 days
                               V
in a  simulated prairie pond microcosm.   In shallow artificial streams, a 50

percent loss of atrazine occurred in 3.2 days  (Kosinski 1984; Moorhead and

Kosinski 1986).  Lay et al.  (1984)  reported an  82 percent loss  in  5 days and a

95 percent loss in 55  days.  The  half-life of - atrazine in wetland  mesocosms

was from 8 to 14 days  (Detenbeck  et al.  1996).   The half-life of "C-labeled

atrazine has been measured in  estuarine  water as 3 to 12 days,  compared to 15

to 30 days in estuarine sediment  and 330 to 385 days  in agricultural soils

(Jones  et  al. 1982;  Kemp et al. 1982a) .

      These rapid losses in small artificial  systems  and in an  estuarine

environment are contrasted with reports  of a  300-day  half-life  in  a larger

lake  system (Yoo and Solomon 1981) , surface water losses of only 33 percent in

120 days and 0 percent in  85 days in two separate 0.49 hectare  pond

applications (Klaassen and Kadoum 1979), and  a  loss of only 40-50  percent in

pond  water over a period of more  than  5  months  (Gunkel 1983) .   In  two months

time, approximately 25-30  percent of individual 20 and 500 ^g/L atrazine

applications to a 0.045 hectare Kansas pond had disappeared  from the water

(deNoyelles et al. 1982).  Approximately 25 percent of the initial

applications remained after  12 months.  The half-life of atrazine  was

approximately 3 months in  Tasmanian streams  (Davies et al. 1994a).

      The  above information  indicates  that  the  persistence of atrazine  in

water is highly variable,  dependent perhaps upon both the nature of  the

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 a
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remained after 2 years.  The authors  concluded that the accumulation and later

release  of atrazine is greatest at cold water temperatures and in sediments

with  lew adsorption capacity.

       The major atrazine degradate in aquatic systems is hydroxyatrazine (U.S.

EPA 2000) .  Others include deethylatrazine, 'deisopropylatrazine, and

diaminoatrazine.  The degradation products  of atrazine were found to be less
                                it
toxic to algae  (Stratton 1984)'and submerged aquatic plants (Jones and

Winchell 1984) then the parent compound.  Equivalent studies of atrazine

degradate toxicity to aquatic animals is sparse.  Results from mammalian

studies  indicate that some atrazine degradates may be more toxic than parent

compound (TJ.S. EPA 2000).

       The mode of atrazine's toxic action toward plants 'is blockage of

electron transport within the Kill reaction of photosystem II, thereby

inhibiting photosynthesis (Moreland 1980) .   Vascular plants and algae are both

affected by this mode of action.  In this way, atrazine has the demonstrated

capacity to reduce primary productivity in  aquatic ecosystems  (deNoyelles et

al. 1982;  Dewey 1986; Herman et al. 198S; Kosinski and Merkle 1984; Pratt et

al. 1988) .  On the other hand,  the mode of  toxic action toward aquatic animals

has not  been documented, probably because atrazine is not considered acutely

toxic to these species.  Recent evidence implicates atrazine as an indirect

endocrine disrupter  (Dodson et al. 1999; Petit et al. 1997) that may act by

stimulating the activity of the aromatase enzyme that converts  testosterone to

estrogen (Sanderson et al. 2000).  Other investigators have demonstrated that

atrazine causes induction of xenobiotic metabolizing systems  (Miota et  al.

1999),  and enhances the .toxicity of.organophosphorous insecticides to  aquatic

invertebrates  (Belden and Lydy 2000; Pape-Lindstrom and Lydy 1997) .

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       Several,reviews exist on atrazine and . its  environmental impact (CCREM




 1989; deNoyelles  et  al.  1994;  Eisler  1989; Kuber 1993,  1994;  Solomon et  al.




 1996).  These  reviews indicated that  a few species of  aquatic plants have been




 shown to be  slightly affected  by atra=ine at concentrations below 10 Mg/L.




 The review by  deNoyelles et al.  (1994) stated that herbicides have little




 direct effects upon  animals, and that they tend  to produce ecosystem effects




 from the bottom of the food chain upward, in contrast  to  insecticides which




 act in the opposite  direction.   Huber (1993) and Solomon  et al.  (1996) stated




 that plants readily  recovered  from the inhibitory effects of  atrazine once the




 exposure was reduced or  eliminated.





   '    Comprehension  of the  "Guidelines for Deriving Numerical National Water




 Quality Criteria for the Protection of Aquatic Organisms  and  Their Uses"




.(Stephan et al. 1985), hereafter referred to as  the Guidelines,  and the.




 response to public comment  (U.S.  EPA 1985) are necessary  -to understand the




 following text, tables,  and calculations.  Results' of  intermediate




 calculations such as  recalculated LC50 values and Species Mean Acute Values




 are  given to four significant  figures to prevent roundoff error in subsequent




 calculations, not to  reflect the precision of values.   The criteria presented




 herein  are the Agency's  best estimate of .maximum concentrations  of the




 chemical of concern  to protect most aquatic organisms  or  their uses from any




unacceptable short- or long-term effects.  Whenever adequately justified, a'




national criterion may be replaced by a site-specific  criterion (U.S. EPA




 1983a),  whi.ch>ay include not only site-specific criterion concentrations




 (U.S. EPA 1983b),  but also  site-specific durations of  averaging periods  and




 site-specific frequencies of allowed excursions  (U.S.  EPA.1991).  The latest




comprehensive literature  search  for, this 'document was  conducted in November,




1999.  Data in the files  of the  U.S.  EPA's Office of Pesticide Programs

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concerning the effects of atrazine on aquatic organisms and  their  uses have



been  evaluated for use in the derivation of aquatic life criteria.   Some more



recent information was also included.






Acute Toxicity to Acraatic Animals






                                i'1
       The data that are available according to the Guidelines  concerning the



acute toxicity of atrazine are presented in Table 1.  Acute  toxicity data for



eight freshwater invertebrate species ranged from 720 ^g/L for first instar



larvae of a midge, Chironomus tentans (Macek et al. 1976} to 49,000  ^g/L for



the cladoceran, Daphnia magna  (Putt 1991).  A hydroid coelenterate (Hydra sp.)



was the second most sensitive invertebrate .tested, with an EC50 of 3,000 ^g/L



(Brooke 1990).  Stonefly  (Acroneuria sp.) nymphs had an LC50 of 6,700 ^g/L and



the asvphipod,  fiyalella azteca, had. an LCSO of 14,700 ^ug/L  (Brooke  1990).  A



cladoceran (Ceriodaphnia duiia) had a Species Mean Acute Value (SMAV) of



>12,120 ^g/L (Jop 1991a; Oris et al. 1991).  The remaining invertebrate



species tested, a snail  (Physa sp.) and an annelid  (imnbz-iculus varieg-atus) ,



had LCSO values in excess of 34,100 and 37,100 ^g/L, respectively  (Brooke


1920) .



       The rainbow trout  (Oncorhynchus mykiss) was the most sensitive



freshwater vertebrate species tested, with an LCSO of 5,300  ^g/L (Beliles and



Scott 1965).  The goldfish, Carassius auratus, was 11.32  times less sensitive



to atrazine (Table 1) .  The fathead minnow  (Pixephales promelas) had a SMAV  of



20,000 jig/L (Dionne 1992), while the LCSO for the brown  trout   (Salmo trutta)



was 27,000 ^g/L  (Grande  et al. 1994).  The SMAVs  for the  remaining vertebrate



species, all fishes, were 6,300; >8,000,  >10,000,  >10,000,  and >18,000



for the brook trout, Salvelinus fontinalis  (Macek et al.  1976); bluegill,

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          macrbciiru*  (Bellies and Scott 1965;  Macek et al. 1976); largemouth  '

 bass,  Micropterua sal^cides (Jones 1962);-channel catfish, Jctalurus punctatus

  (Jones 1962);  and coho salmon,  Oncorhyrichus kisutch (Lorz et al. 1979),

 respectively.   The SMAV was based upon flow-through tests i.n the cases of the

 fathead minnow and bluegill sunfish,  where  other test results were also

 available.            • '
                                 i>                      •
        Freshwater Genus Mean Acute Values were identical to the SMAVs in all

 cases  with  the. exception of Oncorhynchus where the two species tested had

 different SMAVs.   Three of the  four most sensitive freshwater .genera to

 atrazine are invertebrates.  The freshwater Final Acute Value for atrazine was

 calculated  to  be  702.4 ^g/L using the procedure described in the Guidelines

 and the Genus  Mean Acute Values for invertebrates and fish in Table 3.  The

. freshwater  Final  Acute -Value is lower than  all available freshwater Species

 Mean Acute  Values (Figure 1) .

       The acute  toxicity of atrazine  to resident North American saltwater

 au-imai-s has been  determined with eight species of invertebrates and two

 species of  fish  (Table 1) .   Although  only two fish species were tested, fish

 appear to have a  similar sensitivity  to atrazine as do invertebrates.  The

 saltwater Species  Mean :Acute Values range from 2,324 ,ug/L for mysids,

Americanysis. baiaa (formerly Mysidopsis bzhia) ,  to >30/000 ^g/l, for the

 eastern oyster, Grassesfcrea virginica.  The copepod, Aeartia tonsa, had

 similar LC50 values resulting from a  static unmeasured test (Ward and

Ballantine 19'85)  and  two  renewal tests-(Thursby et-al. 1990) with measured

values of 94,. 91.73 and  210.1 pg/L, respectively.  An additional flow-through

measured test  (McNamara  1991a)  with the same  species yielded an LCSO of 4,300

MS/L.   It is unclear  why  there  is  such a, large difference between the  flow-

 through measured value and  the  other  measured results.  There was nothing
                                        10

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unusual  about the variability of the chemistry data from the flow-through

tests  to indicate a problem (coefficient of variations ranged from 2 to 15

percent).   A possible explanation is that the measured values from the static

renewal  tests were conducted with 70 percent technical grade atrazine, while

the  flow-through test used 97.1 percent atrazine.  The other 30 percent may

have contributed to the higher toxicity.  Because there is no obvious problem
                                i"
with the flow-through data set 'for A. tonsa, the Guidelines state that the

flow-through measured value must be used.  Therefore, the SMAV  for this

species  is 4,300 Atg/L.  LC50 values for the ccpepod, Eurytemora. at'finis, were

500, 2,600 and 13,200 Atg/L, at salinities of 5, 15 and 25 g/kg,  respectively

(Eall  et al. 1994a,b).  The resultant SMAV was .2,579 Mg/L-  The opposite trend.

was  observed for the  sheepshead minnow; the LC50 values were 16,200, 2,300 and

2,000  £ig/L at salinities of 5, 15 and 25 g/kg, respectively, for  larval fish

(Eall  et al. 1994a,b).  Two other LC50 values of 13,000 and >16,000 ptg/L for

sheepshead minnow was derived from the flow-through concentration measured

test by  Machado  (1994b) and Ward and Ballantine  (1985).  However, because .the

former LC50 values were from a more sensitive life-stage, an SMAV of 4,208

//gr/L has been calculated for this species.

       Saltwater Genus Mean Acute Values  (Table 3) were  identical  to  the SMAVs

in all cases with the exception of Acartia where the  two  species  tested had

different SMAVs.  Three of the four most sensitive  saltwater genera  to

atrazine are crustaceans.  The saltwater Final Acute  Value  for atrazine,  1,519

pg-/L,  was calculated using the procedure described  in the Guidelines  and  the

Genus  Mean Acute Values in Table 3.  This  saltwater Final Acute Value is  lower

than all available  saltwater  SMAVs  (Figure 2) .      .
                                        11

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 Chronic Toxicity to Aquatic Animals






       The available data concerning the  chronic toxicity of atrazine that  are


 usable according to the •Guidelines are' presented.in Table 2a.   Freshwater


 tests  have been completed with three invertebrate and four fish species.


       The cladoceran,  Ceriodaphnia dubia, was  exposed to atrazine over its


 entire life cycle in two 7-day tests (Oris  et  al.  1991).  The  end result was


 identical in both tests,  with chronic limits of 2,500 and 5,000 ^g/L,  and  a


 calculated chronic value (geometric mean) of 3,536 M9/L.  An accompanying


 acute  toxicity test resulted in an LC50  of  >30,000 ^g/L (Oris  et al.  1991).


 The resultant acute-chronic ratio  was >8.484.


       In  another 7-day life cycle  exposure  with Ceriodaphnia dubia (Jop


 1.99Ib) , atrazine did not affect survival at any of the test concentrations


 (i.e.,  290,  600,  1,200,  2,500 or 4,900 ^g/L).   However,  reproduction was


 significantly reduced  at the two highest treatment levels.  An average of  10


young  per female were  produced at  these  two treatments compared to a mean  of


 23 for the pooled controls.   The chronic limits in this study  were 1,200 and


 2,500  ^g/L.  and the chronic value  was 1,732 ^g/t.   An accompanying acute value


of >4,900  ££g/L (Jop 1991a)  resulted in an acute-chronic ratio  of >2.829.

                 it
Therefore,  the species mean acute-chronic ratio is >4.899 (Table 3).


       The  cladoceran,  Daphnia magna,  was continuously exposed  to atrazine  over


 three  generations for  a  total of 64 days (Macek et al. 1976).   Mean measured


concentrations of 250, 550  and 1,150 Mg/L significantly reduced young


production in the first  generation animals, while production at mean


concentrations of 60 and 140 /J.g/1*  was similar  to controls.  Variability in the


data for  second and third generations precluded statistical significance of


generally  reduced young  production with  increased atrazine concentration.
                                       12

-------
 Chronic limits based on young production by first generation daphnids were  140




 and 250 Aig/L, with"a resultant chronic value (geometric mean) of 187.1 ^g/L.




 A  corresponding acute value of 6,900 ^g/L (Macek et al.  1976} yielded an




 acute-chronic ratio of 36.88.





       The midge, Chironomus tentans,  was continuously exposed to atrazine for




 two generations in a life-cycle test (Macek et al.  1976).   The test was




 initiated by exposing first generation eggs 'through the various larval instar




 stages,  pupation and emergence.  Eggs from first generation adults were then




 continuously exposed in a similar fashion.   Mean measured concentrations were




 110, 230,  420,  780 and 1,330 pig/L.   No significant  differences between




 controls and the lowest exposure (110 ^g/L)  were noted in hatchability,




 survival,  pupation or emergence in first generation animals.  Significant




 reductions in the number of adults  emerging in the  first generation exposure




 occurred at atrazine concentrations of 230  and 420  ,ug/L.  First generation




 larvae  exposed to higher concentrations experienced high mortality at the




 early instar stages.  In the second generation,  hatchability was reduced at .




 420 Aig/L,  while pupation and emergence were reduced at 230 and 420 ^g/L of




 atrazine.   Exposure to 110 ^g/L had no effect on growth or development of the




 chironomid larvae.  Based on these  observations,  the chronic limits were 110




 and 230  ^g/L,  and the resultant chronic value (geometric mean) was 159.1 ^g/L.




A corresponding acute value of 720  /zg/L (Macek et al.  1976) yielded an acute-




 chronic  ratio of 4.525 for Chironomus tentans.




      Rainbow trout (Oncorhynchus mykiss)  were exposed to atrazine in an




 early-life stage test (ELS)  conducted in reconstituted water with a hardness




 of 50 mg/L as calcium carbonate (Whale et al. 1994) .  The ELS test was divided.




 into 3 main stages:  (I)  immediately post-fertilization to hatching (30-day




 duration), '(II)  post-hatch to swim up (28-day duration),  (III) post-swim up to
                                       13

-------
 3 months old (28-day duration),  for  a  total  exposure of 86 days. . Mean




 measured concentrations (mean ±  SD)  were  <10  (water control) , <10 (solvent




 control),  36  ±;12,  130 ± 50,  410 ± 170, 1,100  ±  660,  and 3,800 ± 2,200^g/L,




 respectively.   Significant mortalities  (53.8 percent)  occurred in the highest




 atrazine exposure during stage I and II of the' test although no other dose




 response relationships could  be  defined.  Significant decrease in fish-wet




 weight was observed in. concentrations of  1,100 and 3,800 ^g/L compared to the




 solvent control,  although fry exposed to  1,100 Mg/L did show signs of a




 recovery in wet weight toward the end of  the stage III exposure.  Statistical




 analysis, of the dry weights of these same fish samples showed that a




 significant decrease in weight occurred only in  fish exposed to 3,800 pg/L




 atrazine.  Because  of the recovery in growth at  the 1,100 ^g/L atrazine




 concentration, the  chronic limits in this study  were set at 1,100 and 3,800




 A*g/I.,  resulting in  a chronic  value of 2,045 Mg/L-   An accompanying acute value




 is not available  for this  species, therefore, an acute-chronic ratio'cannot be



 calculated.





       Yearling brook trout (Salvellnus • fontiualis)  and their offspring were




 continuously exposed to  atrazine for 306 days at mean measured concentrations




 of 55,  120, 240,  450  and 720  Mg/L (Macek et al.  1976).  At 90 days,




 significant reductions  in  weight and total length  of first generation fish




 occurred at concentrations of  240 ^g/L and above.   At 306 days, weight and




 total  length 'of first generation fish were significantly less than controls at




 atrazine exposures  of 120  W/L and above.  Fish  at these exposures also




 appeared lethargic  in comparison to-  the controls and fish at 65 ^g/L.




 Spawning activity and hatchability of second generation fry did not  appear to




be affected,  although considerable variability between replicates in the




parameters  of total  number of  eggs spawned, number of eggs per female, percent
                                        14

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fertilisation and hatchabiiity precluded  statistical interpretation.  High


replicate  variability was also observed in morphological development of the


embryos.   At 30 days of exposure,  fry survival was similar for all treatments,


but was  significantly reduced at concentrations of 240 ^g/L and above after


both 60  and 90 days.  As in first generation fry,  length and weight of second


generation fry at 90 days were significantly less  than controls at atrazine
                                ,i
exposures  of 240 Aig/L and above.  Based on the most sensitive measure, i.e.,


growth of  first generation fish at 306 days,  the chronic limits were 65 and


120 pg/L,  with a resultant chronic value  of 88.32  A^g/L-  A corresponding acute


value of 6,300 ^g/L (Macek et al.  1976) yielded an acute-chronic ratio of

71.33 for  brook trout.


      Fathead minnows (P. prcmelas) were  continuously exposed to atrazine for


43 weeks starting with 18-day old fish, continuing through -spawning and into a

second generation for 60 days (Macek et al. 1976) .  Mean measured atrazine


concentrations were 15, 33, 54, 112 and 213 jug/L.   Control fish survival was


low  (53.5  percent) for the first 60 days  of exposure, but after thinning at  60

days, control survival was 93 percent for the remaining 34 weeks.  No


significant differences in survival or growth were observed between weeks 9


and 43 in  first generation fish.  A lack  of spawning activity at 33 and 213
                &
A^g/L was thought to be an artifact of different sex ratios and not due to

treatment.  Otherwise, there were no significant differences in the


reproductive parameters of mean number of eggs produced per  female and mean

number of  eggs per spawn, or in the haichability, survival or growth  of second

generation fish through  30 and  60  days.  Based on a  lack  of  any observed


adverse  effect at the highest exposure of 213 ng/L, and an observation of  25

percent  mortality in  3 to  5-day old fry,after  96 hours of exposure  at 870

       the  chronic limits were set  at 213  and  870 A^g/L,  with  a  resultant
                                        IS

-------
 chronic value (geometric mean)  of  430.5 ^g/L.  A corresponding acute value of




 15,000 pg/L  from Macek et al.  (1976) yielded an acute-chronic ratio of  34.84.




       A fathead minnow full life-cycle chronic test  that  extended for 274 days




 was performed,  with mean measured  atrazine concentrations  of  0,  150,  250, 460,




 990 and 2,000 Mg/L (Dionne 1992).  At 30 days, first generation larval  length




 was significantly reduced by concentrations *990 Mg/L, whereas,  at 60 days,




 length was reduced at  concentrations *460 ,,g/L.  At 274 days,  survival  was




 significantly reduced  at 990 and 2,000 Mg/L of atrazine.   There was no  effect




 upon the reproductive  parameters of number of eggs per spawn,  total number of




 eggs produced, number  of spawns .per female, or number of eggs per female at'




 any treatment level.   Hatching success was slightly, but significantly,




 reduced at concentrations of 250 W/L and above.   Second generation larval




 growth (length and weight) was significantly reduced at. *460  ^g/L of atrazine.




 The chronic limits were  reported to be 250 and 460 jzg/L, based upon first and




 second generation larval, hatching and growth.   This resulted  in a chronic




 value  of  339.1 /^g/L.  An accompanying .acute value of 20,000 ^g/L (Dionne 1992)




 yielded an acute-chronic ratio of 58.98.





       Bluegills   (Lepomis macrochirus)  were continuously exposed to atrazine




 for 18  months starting with 7-10 cm long  fish,  continuing  through spawning,




 and into a second generation  for 60 days  (Macek et al. 1976).   Mean measured




 exposure concentrations were  8,  14, 25, 49 and 95 MS/L.  Survival and growth




of  first generation fish exposed to atrazine for 6 and 18  months were similar




 to  the controls.   Spawning activity was' too sporadic to indicate any adverse




 effects.   Percent hatchability of eggs  was similar to controls at    '




concentrations between 14 and  95 MS/L.  ' Low fry survival in the second




generation controls for the first 30 days precluded observations on survival




effects due to atrazine in this  time interval.   However, between 30 and 90
                                       16

-------
days,  survival was near 100 percent in the  controls and all atrazine




treatments.   Total length of second generation fish through 90 days was




considered to be unaffected by any of the atrasine exposures.  From a lack of




any adverse  effect at concentrations as high as 95 ^g/L and from an observed




loss of  equilibrium in bluegills exposed to 500 ngfL for 28 days, the chronic




limits were  set at 95 and 500 ^tg/L.  The resultant chronic value was 217.9




^g/L.  A corresponding, acute vaiue of >8,000 yug/L  (Macek et al. 1976) yielded




an acute-chronic ratio of >36.71.




       The acute values for D. magna, C. tentans, S. foo.tina.lis, P. promelas




and £. macrochirus in tests reported by Macek et al. (1976) were used in




calculating  acute-chronic ratios even though -the acute test concentrations




were not measured.  This was because of close agreement between nominal and




measured concentrations in the chronic tests.  For six chronic tests, the




overall  agreement between measured and nominal concentrations was 94.4




percent.  Therefore, it appeared likely that the nominal concentrations




presented for acute tests were also in good agreement with actual




concentrations.




       The chronic toxicity of atrazine to saltwater species  has  been




determined in three 8-day life cycle  tests with  the copepbd,  Eurytemora




a-ffinis, a 28-day life cycle test with the mysid,  Americamysis bahia,  and an




early  life-stage test  (28-day) with  the sheepshead minnow, Cyprinodon




vajries-atus  (Table 2a) .  Survival was  the most  sensitive  endpoint in the  8-day




chronic  tests with E. af finis.  Tests were performed at  salinity levels  of.  5,




15 and 25 g/kg.  At a salinity of  5  g/kg,  survival was  significantly reduced




to 37  percent at the  17,500  ^g/L  concentration,  while  at the next lower




concentration of 12,250 Mg/L it  was  similar  to controls  at 71 percent (Hall et




al.  1995) .  The chronic value was  14,640 ^g/L.   At a  salinity of 15 g/kg, the
                                        17

-------
chronic  limits were 17,500 and 25,000 jzg/L, and the chronic value was  20,920

Aig/L.  Sensitivity appeared greater at a salinity of  25  g/kg,  with chronic

limits of 4,200 and 6,000 ^g/L, and a chronic value of  5,020 f^g/l,.  Only at

this  highest salinity level was the.acute value greate'r  than, the chronic

value.   The resultant Acute-Chronic Ratio of 2.629, determined at a salinity

of 25 g/kg (13,200 ^g/L -=- 5,020 ^g/L) , was considered to be the correct ratio
                                ii
for this species, and was used'in subsequent calculations involving the

Species  Mean Acute-Chronic Ratio.

      Sur-vival was the most sensitive endpoint in the mysid test  (Ward and

Ballantine 1985).  Survival was 60, 30, and. 2-Q percent  at 190, 290 and 470

ptg/L, respectively.  No statistically significant effect was observed for

s-ur-vival at concentrations S80 /ig/L.' Reproduction did.  not occur at 470 pg/L,

but no adverse effects on reproduction were observed, at  all lower

concentrations.  The chronic value for mysids, based  on survival, is 123.3

ptg/L.  The acute value, as determined by the same authors, is 1,000 fJ.g/1,  a=.d

the resulting acute-chronic ratio is 8.110  (Table 2b).

      In the sheepshead minnow test  (Ward and. Ballantine 19B5), juvenile

survival was;.significantly reduced at 3,400 ptg/L, but not at  <;1,900 M9/L.   All

fish  exposed to 5,700 ^g/L died.  There was no effect on either hatching

success  or- growth in any of the concentrations with  surviving fish  (s5,700

Atg/IO .   The chronic value for sheepshead minnows, based on mortality of

juveniles, is 2,542 Mg/L •  The acute value  for the  sheepshead minnow, as

determined by the same authors, is a  "greater than"  value  (>1S,000  jug/L) .

Therefore, the resulting acute-chronic value  is  >S.294.

      The range of species mean acute-chronic ratios for both freshwater and

saltwater differ by more than a factor of  10  (Table 2b) ,  and  are  not  related

to rank,  order of acute sensitivity (Table  3) .  For both  freshwater  and

-------
saltwater,  separate fish and invertebrate chronic values were  calculated using




separate fish and invertebrate Acute-Chronic Ratios  (Table  3).   Acute-Chronic




Ratios  with greater than values were not used for these calculations.  The




freshwater Fish Chronic Value of 12.35 ^g/L is the quotient of  the Final Acute




Value  (702.4 pg/L) and the Fish Final Acute-Chronic Ratio  (56.86).  Likewise,




the Invertebrate Chronic Value of 90.95 W/L is the quotient of  the Final




Acute Value (702.4 Mg/L)  and the' Invertebrate Final Acute-Chronic Ratio




(7.723).   The Final freshwater Chronic Value is 12.35 j/g/L, the  lesser of the



two chronic values.





      The saltwater Fish Chronic Value of 26.71 Mg/L is the quotient of the




Final Acute Value (1,519 Mg/L)  and the Fish Final Acute-Chronic  Ratio  (56.86).




Likewise,  the Invertebrate Chronic Value of 196.7 Mg/L is  the  quotient of the




Final Acute Value (1,519 Mg/L)  and the lnvertebrate Final Acute-Chronic Ratio




(7.723).   The Final Saltwater Chronic Value is 26.71 ^g/L,  the  lesser of the




two chronic values.









Toxicitv to Aquatic Plants        • .                        .










      For inclusion in Table 4, according .to the Guidelines, exposures with




algae must have been for a minimum of 4 days.  With vascular plants, chronic




exposures must have been conducted.   In both cases, it is  a requirement'that




the concentrations of atrazine were measured during the tests.   A Final Plant




Value can be obtained by selecting the-lowest result from- a test with an




important aquatic species in which the concentrations of  test  material were




measured and the endpoint was biologically important.




      Two species of freshwater green algae were exposed  to atrazine in




studies in which the exposure duration was 4 days or longer and the atrasine
                                       19

-------
 concentrations were measured  (Table 4).  C^lamydomon./reinh.«ieii  cell




 numbers were reduced 50 percent after 4 days of exposure  to  51  ^g/L,  after 7




 days  of exposure to 21 ^g/L, and after 10 dayg Qf exposure fco 1Q>2  ^





 (Schafer et;al. 1993).  The -no observable effect concentrations"  (NOECs) were




 3.4,  5.1 and 3.7 pg/j, at 4, 7 and 10 days,  respectively  (Schafer et al. 1994).




        Selenastrum '• c.prico™,tu» had ' a 4-day ECSO of 4 ^g/L,  based upon cell




 numbers (University of Mississippi 1990) .   The EC50 values for *pheophytin a




 and chlorophyll a content were 20 and 150  W/I|. respectively.   Using  the ,a»




 species and cell number as an endpoint, *ala and Giesy (1990) reported a 4-day




 ECSO of 128.2  Mg/L,  and Eoberg- (199la)  reported a 4-day ECSO of 130 w/i,.




 Hoberg  U993a)  calculated a 5-day ECSO  of  55 w/l.   EC10 values at  .4  and 5




 days were  sol and 26  Mg/L,  respectively, whereas,  EC90 values at 4 and 5 days




 were 190 and 120 ^g/L,,  respectively (Hoberg 1991a,  I993a) .





       A 7-day  exposure  of  the  duckweed, Lemna g±bba,,to atrazine resulted in,




 an ECSO of  180  Mg/L, based upon  frond production (Hoberg I99lb)  .  Two 14-day.




 studies were also conducted with L. yiiba  (Hoberg i993b,c).  A  major




 difference  in  these  two  studies  was that, in the  latter study,   the effect




 concentrations  were  calculated based upon the atrazine concentrations that   •




 were measured,on the last  day only.  This may have  resulted in  effect levels




 that appeared to be  lower  than in  the first  study,  where concentrations were




 measured more often during  the test.  In the first  study (Hoberg I993b) , using




 frond  number as an endpoint, the EC10, ECSO  and EC90  values were 6.2,  37, and




 220  izg/r,, respectively, after 14 days of exposure.  Using frond biomass, the




 EC10,  ECSO and EC90 values were  12, 45 and  170  w/.t.  respectively.  In the




 second study (Hoberg 1993c) , the EC10, ECSO  and EC90  values, were 2.2,  50, and




 98 Mg/L, respectively, using the frond number endpoint,  while the respective




values  for^frond biomass were 4.2, 22, and  110
                                      '  20

-------
      Exposure of a different species  of  duckweed,  Lemna minor, to atrazine

for 14  days resulted in a NOEC of  10 ^g/L based upon a biomass endpoint

(University of Mississippi 1990).   In  this study,  "lowest observable effect

concentrations" (LOECs)  of 10 and  100  M9/L were obtained for the endpoints of

mature  frond production and biomass, respectively.   The EC50, based on

biomass,  was 8,700 pg/L.  In another study using L. minor (Kirby and Sheahan
                                11
1994) ,  10-day exposures to atrazine yielded EC50 values that were comparable

to those  found for L. ffibba by Hoberg  (1993b,c).  EC50 values of 56, 60 and  62

Atg/L were obtained based upon frond number,  fresh weight and chlorophyll

content,  respectively.

      Slodea (Slodea. canadensis) was exposed to atrazine both in the absence

and presence of sediment (University of Mississippi 1990) .   In the absence of

sediment,  LOEC values of 10 and 100 M9/L  were observed, based upon mature

frond production and biomass, respectively.   With sediment present, the

biomass LOEC was also 100 ;zg/L. Bicmass  SCSO values were 1,200 and 25,400

^g/L whea sediment was absent and  present, respectively, in the test systems.

A freshwater Final Plant Value was not calculated,  as none of the species -

tested  met the Guidelines criteria for such a determination.

      Information on the sensitivities of saltwater plants to atrazine is

available for five phytoplankton species  and four vascular plant species,

representing eight genera  (Table 4) .   Although the phytoplankton test results

do not  meet the minimum retirement of a  four-day exposure, they are included

here  to show that their sensitivity ta atrazine is similar to vascular plants.

All of  tha plant effect concentrations were less than  the acute values for

aquatic animals.  Short-term  (two and three day) growth  tests with

phytoplankton resulted  in EC50 values ranging  from 79  to 265 A£g/L  (Mayer  1987;

Walsh. 1933); a factor of only 3.4.  Two species of estuarine submerged
                                        21

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 vascular plants,  Potamogreton perfollatus  and Myriophyllum spicatum,  expoaed


 for 28-35 days  to various concentrations  of atrazine, had IC50  values for


 final biomass and photosynthesis between  25 and 117 ^g/L, with  the biomass


 endpoint being  more sensitive in both species (Kemp et al. 1982b<, 1983, 1935).


 The sago pondweed,  Potzmogeton pectinatus,  was  tested (Hall  et  al. 1997) for


 atrazine toxicity for 28 days at three salinities (1, 6, and 12  g/kg) .  Dry
                                 it

 weight was  the  most sensitive endpoint with chronic values of 21.2,  21.2 and  -


 10.6 Azg/L at salinities of 1,  6, and 12 g/kg salinity, respectively.  A two-


 way ANOVA yielded a chronic value of 5.3  pg/L of atrazine.   Four separate 21-


 day exposures of  the seagrass,  Zostera marina^  resulted in LC50  values ranging


 from 100 to 540 Mg/L (Delistraty and Hershner "1984) .   A saltwater Final Plant


 Value was not calculated,  as none of the  species tested met  the-Guidelines


 criteria for,-such a determination.





'Bioaceuznulation.






       The data available according to the Guidelines  concerning  the


 bioaccumulation of  atrazine are included  in Table 5.   Only freshwater data are


 available.   Macek et al. (1976)  analyzed  muscle tissue or the eviscerated


 carcasses of fish at the end of extended  exposure periods.   Brook trout
             I                                  .

 exposed to atrazine at  740 Mg/L for 308 days contained less  than 200 ^g/kg of


 atrazine in muscle  tissue,  resulting in a bioconcentration factor  (BCF) of


 <0.27,   Fathead minnows exposed to atrazine at  210 jug/L for  301  days had less


 than 1,700 M9/kg  of atrazine in pooled samples  of eviscerated carcasses, for a


 BCF of  <8.1.  Bluegills exposed to 94 jug/L  for  546 days also contained less


 than 200 ^g/kg in their muscle  tissue,  for  a BCF of <2.'l.
                                        22

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      Dionne (1992) exposed fathead minnows  to atrazine for up to 274 days

using uC-labeled atrazine and measuring the  radiolabel in fish tissue.  The

values obtained represent maximum possible BCFs.   Regardless of the life-stage

or exposure duration, maximum BCFs were less than or equal to 8.5 in all
cases.
      There is no U.S. Food and Drug Administration action level or any other
                                11
established maximum allowable concentration of chemical residues in tissue

available  for atrazine.  Therefore,  a. Final Residue Value cannot be

determined.
Other Data
      Many tests with atrazine and various  freshwater or saltwater organisms

have been conducted.either for a different  duration or by different protocols

than those specified in the Guidelines  for  inclusion in Tables 1, 2, 4 and 5.

These test results are presented in Table  6.   For example, plant tests were

included in Table 6 rather than Table 4 if  the test duration was less than 4

days or  the exposure concentrations were not  measured.  Tests with animals

were included in Table 6 for a number of reasons, including considerations of

test duration,  type of test, and test endpoints other than those of toxicity

or bioaccumulation.  Below is a summary of  their results.

      At the lowest levels of biological organization, mixed nitrifying

bacteria were unaffected regarding ammonium oxidation at 28-day exposures up

to 2,000 ^g/L of atrazine  (Gadkari 1988),  and cell growth in the bacterium,

Pseudomonas putida, was not inhibited following a 16-hour exposure at 10,000

Atg/L  (Bringmann and Kuhn 1976, 1977).  Progressing phylogenetically, Rohwer

and Fluckiger (1979)  obtained a 14-day growth LOEC of 2,160 ^g/L  for Anaiaena
                                       23

-------
 cylindrica, while Stratton (1984)  obtained  a  12  to 14-day EC50 of 1,200 Mg/L

 in terms of cell  number.  The  latter EC50 value  was approximately 5 to 7 times

 higher than the 24-hour EC50 values based on  »C uptake of 253, 178 and 182

 ^g/L as reported  by Larsen et  al.  (1986) for  this same species (Table 6).  The

 other species  of  cyanobacteria tested by Stratton (1984), Anabaena inaequalls

 and Anahaena.vajriaiills, had highly different EC50 values of 30 and 4,000 Mg/L '
                                 i'
 after 14 days.  A.  inaegualis  and  PseudoanaJbaana 'sp.  exhibited reduced

 photosynthetic uptake of "c in the amounts  of 65 and 91 percent, respectively,

 following, a 22-hour exposure to 2,667 ^zg/L  of atrazine (Peterson et al. 1994).

       A number of  tests have been  performed with the  cyanobacterium, AnaJbaena

 r-los-aguae. . Hughes (1986)  and Hughes et al.  (1986,  1988) reported an ECS 0

 based on cell number of 230 ^g/L following  a  5-day exposure.  A concentration

 of  40 ^g/L non-radiolabeled atrazine reduced  "C uptake by approximately  50

 percent after 1- to  3  days  of exposure, after  which the reduction was less

 (Aboxi-Waly et al.  1991a).  At  this concentration of atrazine, chlorophyll a

 content was initially reduced  but  recovered with time.  Using this parameter,

 the  3-day EC50 was  58  ^g/L, while  the 7-day EC50 was  76S ^g/L  (Abou-Waly

 1991b) .   A. flos-acyuae- had a 4-day EC50 based on chlorophyll a that exceeded

 3,000 ^g/L  in a study by Fairchild et al.  (1998).

       The cyanobacterium Aficrocystis aerug-inosa  exhibited the onset of cell

growth inhibition at  a concentration of 3 ^g/L in an 8-day exposure  (3ringmann

and  Kuhn 1976, 1978a,b) .   After 5  days of exposure,  cell numbers were

 significantly reduced at 108 /ig/L, and-the  minimum algistatic concentration

was  440  ^g/I. (Parrish 1978) .   Kallqvist and Romstad  (1994) obtained a  6-day

EC50  of  630 ^g/L with M. aeruginosa., while  Peterson et al.  (1994) reported

 that  photosynthetie "C uptake  was  highly reduced  (84-96" percent)  in M.

aerug-inosa  following  a 22-hour exposure to  2,667 Mg/L of atrarine.  A  4-day
                                        24

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EC50 of  90 pg/L was reported  for an unidentified  species of Microcystis based


on biomass (Fairchild  et al.  1998) .


       Toxicity studies of  atrazine toward several other species  of


cyanobacteria have been reported.  Peterson  et  al.  (1994) found  that


Aphan±zaaenon flos-aguae and  OscilJatoria sp. exhibited highly reduced


photosynthetic uptake  of "C (97  and  87 percent, respectively)  from a  22-hour
                                n
exposure to 2,657 ^g/L of  atrazine.  The latter is consistent with  the lowest


complete inhibition of growth reported for Oscillatoria cf. chalybea.  after 6


days of  exposure to 2,160  ptg/L atrazine  (Schrader et al. 1997).   A 31-day


exposure of Plectonema ijoryanura  to 10,000 ^g/L  of atrazine  resulted in a 69


percent  decrease in cell numbers  (Mallison and  Cannon 1984) , whereas, S-day


exposures of Syneciococcus leopolensis yielded  an EC50  of  130  IJ.g/1> (Kallcvist

and Romstad 1994).


       The green alga,  Ankistradesmus Jbraunii, had an 11-day EC50 of 60 jJ.g/l>


 (Burrell et al. 1985) .. Similarly, 14C  uptake EC50 values of 72 and 61 jUg/L


resulted from 24-hour  exposures  of Ankistirodesmus sp. to  atrazine (Larsen  et


al. 1986) . The green  alga, Chlamydcinonas geitleri Ettl,  had a slightly higher


SC50 of  311 A^g/L based on  C02 fixation after a  1-hour exposure  (Francois and


Robinson. 1990) .  Similarly, a growth-based EC50 of 330  £tg/L was obtained for

                S,
Chlaaydomorjas noctig-ajna after 3  days of  atrazine exposure (Kallqvist and

Romstad 1994).


       The green  alga,  Chlamydomonas  reinhajrdtii, appears more sensitive to


 atrazine, exhibiting approximately a 3-2  percent  inhibition of photosynthesis


 in an. 8-hour  exposure to 10 M3/L  (Valentine and  Bingham 1976) ,  and EC50 values


 based on reduction in photosynthetic activity  (14C uptake)  in-24-hour exposures


 of 19  to 43 Aig/L of atrazine  (Larsen et al. 1986) .   Atrazine-sensitive and


 atrazine-resistant strains of C. reinha.rdt.ii responded to  2-minute exposures
                                         25

-------
 by a difference of approximately an order of magnitude in  their respective

 EC50 values of 45 and 484 ^g/L  (Hersh and Crumpton 1989) .  A  65-hour exposure

 to 49.6 ^g/L resulted 'in a 13 percent reduction of chlorophyll  (Hiranpradit

 and Foy 1992), and Fairchild et al. (1998)  obtained a 96-hour chlorophyll-

 based EC50 of 176 ^g/L for this same species.

       Foy and Hiranpradit (1977) exposed an unknown Chlamydomonas  sp. to
                                11
 various concentrations of atrazine for 72 to 96 hours.  Concentrations of 50

 to 52  Mg/L inhibited growth by 84.9 percent and reduced chlorophyll by 12.8

 percent.   Slight additional increases  in growth inhibition were observed with

 increased atrazine concentrations up to 832 ^g/L,   Fairchild  et al. (1994a)

 obtained  a 4-day EC50 based on biomass of 176 ^g/L to a different  species of

 Chlasnydomonas.         •       .   .               ,

       Chlorella  fusca cell reproduction was reduced and an EC50 of 26 fJ.g/L was

.calculated following a 24-hour exposure to atrazine (Altenburger et al. 1990).

 Similarly,  Faust et al.  (1993)  obtained a 24-hour  EC50 of 15  Mg/L  for this

 species,  and Kotrikla et al.  (1997) report 14-day  EC50 values based on growth

 inhibition of 53.91 (exponential growth phase)  and 75.73 f^g/Ij  (stationary-

growth phase).   In contrast,  Chlorella kessleri exhibited 30  percent growth

inhibition following a 72-hour exposure at a concentration of 1,078 M9/L  (El-

Sheekh et  al.  1994),  while C.  pyrenoidosa had 70 to 95 percent  reduced growth

 following  2-week exposures to atrazine concentrations ranging from 500 to

10,000  Mg/L  (Virmani et al.  1975).   Photosynthesis in this species was

inhibited  by approximately 64 percent  following an 8-hour exposure to 100 pg/I,

atrazine  (Valentine and Bingham 1976) .  Stratton (1984) obtained an EC50 of

300 ^ug/L  following a 12- to 14-day exposure.  A 30 percent reduction in growth

and 40  percent reduction in chlorophyll a was observed in a 10-day exposure to

53.9 ^g/L  (Gonzalez-Murua  et al. 1985),  while a 110-hour exposure  to 49.6 /zg/L
                                       26

-------
reduced chlorophyll  by 39  percent  (Hiranpradit  and  Foy 1992) .   Photosynthetic

CO3 uptake was inhibited by more than 80 percent in C. pyrenoidosa  following a

less than  50-minute  exposure  to  125 M9/L  (Hannan  1995).

      The  green  alga,  Chlorella  vulgaris, had 24-hour EC50 values of 325,  305

and 293 ^g/L  in  three  separate tests based upon "c  uptake (Larsen et al.

1986).  Similarly, ,a 30-minute EC50 value of 305  /zg/L based on decreased
                               n
oxygen evolution was obtained for  the aame species  by Van der  Heever and

Grobbelaar (1997).   Following 7  days of exposure  to 250  to 5,000 ptg/L (only

2.3 to 4.7 percent remained on day 7), dry weights  of C.  vulgaris were reduced

from 31 to 62 percent  (Veber  et  al. 1981).  This  same species  had an EC5Q. of

94 ^g/L based upon chlorophyll concentration after  a 96-hour exposure

(Fairchild et al. 1998) .   Reduced  growth was initially observed for C.

vulg-aris exposed for 12  days  to  10 ng/I*. although signs  of recovery were

evident by the end of  the  exposure (Berard et al. 1999) .

      In an undefined  species of Chlorella, a 72- to 96-hour atrazine exposure

at 52 M9/L resulted  in a 31 percent inhibition  of. growth and a 39 percent

reduction  in  chlorophyll (Foy and  Hiranpradit 1977) .   In that  same study,

higher exposures generally resulted in greater  adverse effects.  More.

recently,  a 2- to 3-day atrazine exposure of 21.6 ^g/L reduced the growth rate

of one Chlorella sp. by 55 percent (Hersh and Crumpton 1987) ,  and another

study using Chlorella  sp.  exhibited very rapid  responses to atrazine with EC50

values of  35  to  41 ^g/L based upon photo synthetic oxygen evolution following  a

2-minute atrazine exposure (Hersh  and Crumpton  1989) .  Fairchild et al.

(1994a) reported a  4-day biomass-based EC50 of  92 /zg/L in yet another study

using an unidentified  species of the genus Chlorella.

      Virmani et al. (1975)  observed 75 and 92  percent reductions in growth of

a much less  sensitive  species of green  algae, Chlorococcum' hypnosporum,
                                       27

-------
 following 2-week exposures to 5,000 and 10,000 M9/L atraziiie,  respectively.

 Similarly,  a high test concentration {2,157 M9/D was necessary, to  inhibit

 calcification in Gloetaenium loitlesbergarianum in a 96-hour'test  (Prasad and

 Chowdary 1981).  Short exposures (2 minutes)  to Franceia sp.  yielded EC50

 values  between 430 and.774 jzg/L, measured as photosynthetic oxygen  evolution

 (Hersh  and Crumpton 1989).
                                i >
      In three tests with the green alga,  Scenedesmus obliquus,  the 24-hour

 EC50 values for "c uptake were between 38 and 57 p;g/L  (Larsen et al. 1986).

 The green alga, Scenedesmus guadricauda,  exhibited photosynthesis inhibition

 of approximately 42 percent after 8 hours  at an atrazine exposure of 10 pg/L

 (Valentine  and' Bingham 1976).  Bringmann and Kuhn (1977, 1978a,b) found that

 30 Atg/L caused the onset of cell multiplication inhibition after 8  days of

 atrazine exposure to this species.   S.  quadricauda exhibited  a 12-  to 14-day

 EC50 of 100 Mg/L based on cell number (Stratton 1984) .  Bogacka et  al.  (1990)

 studied photosynthesis reductions in S.  quadri-cauda at various concentrations

 after 8 days of atrazine exposure.   These  authors observed a  gradation from

 4.5 percent reduction at 4 M9/L to a 99.3  percent reduction at 337  Mg/L-

 Similarly,  photosynthetic 14C uptake was highly inhibited  (96  percent) after  22

horrs at 2,667 Atg/L of atrazine (Peterson et al. 1994).  This species had a

 96-rhour EC50 of- 169 jug/L, based upon chlorophyll concentration (Fairchild et

al. 1998) .

      In this same genera of algae, Scenedesmus suispicatus had a 4-day EC50

of 110  Mg/L (Geyer et al. 1985), and Schafer et al.  (1994)  found that 37 p:g/L

of atrazine inhibited the effective photosynthetic rate of  this species by

 57.4 percent within 24 hours.  This latter apparent effect  concentration was

 corroborated- by Kirby and Sheahan  (1994)  who reported  a 2-day EC50  of 21 jug/L

based on cell numbers, as well as Zagorc-Koncan  (1996) who  reported a 24-hour
                                       28

-------
 EC50 value of, 25 pg/L based on net assimilation and inhibition.  Reinhold et

 al.  (1994) observed a 50 percent reduction in dry mass at 21.5 f^g/L within 24

 hours,  and Behra et al.  (1999) reported a 60-day NOEC based on growth and

 photosynthetic oxygen evolution for this species of 20 ^g/L.

       Exposure of an unidentified species of  Scenedesmus for 72 to 96 hours at

 50 pg/L resulted in 60.2 percent growth inhibition (Foy and Hiranpradit 1977),
                                11
 and  increased concentrations resulted in increased growth inhibition.

 Pairchild et al. (1994a) obtained a 4-day EC50 based on biomass of 169 Mg/L.

       The green alga,  Selenastrum capricorzmtum,  exhibited a significant

 reduction in cell numbers following a 5-day exposure to 54 ^g/L of atrazine

 (Parrish 1978) .   In this study,  chlorophyll a reduction increased as

 concentrations increased from 32 and 200 M9/L-   The minimum algistatic

 concentration was determined to be 200 ^g/L.   A similar 5-day LOEC for S.

 capricornutum growth of 220 //g/L was recently reported by Schrader et al.

 (1998).   Interestingly,  a 7-day exposure at 100 ^g/L resulted in a 13.8

percent  increase in biomass, whereas 1,000 ^g/L resulted in decreases (Johnson

1986) .   The  lowest complete inhibition concentration of growth after a 6-day

exposure was 2,160 /ig/L (Schrader et al.  1997).

      There  are  a number of additional EC50 values from exposures of S.

capricornutum to atrazine (Table 6).  Larsen  et al. (1986) obtained 24-hour,

EC50 values  of 53,  34  and 42 ^g/L based upon  "c uptake.  In a couple of 21-day

exposures (Turbak et al. 1986),  biomass-based EC50 values of 58.7 and 410 ,ug/L

were obtained using algal assay media and creek water for test media,

respectively.  Likewise, EC50 values were 69.7 and 854 ;ug/L, respectively,

using these  two  media  in 24-hour tests that measured photosynthetic oxygen

evolution.   Roberts et al.  (1990)  reported 5-day EC50 values of 100 and 95

     based on cell numbers,  arid an EC50 of 50 jug/L based on -cell numbers was
                                       29

-------
 reported in a 4-day exposure by Versteeg (1990).  Similarly, El Jay et al.

 (1997), found the 4-day 1C50 values based on chlorophyll a content  to be 80

 ^g/L.   Reductions in chlorophyll content and in "c uptake occurred at 130

 in 1-  to 7-day exposures  (Abou-Waly et al .  1991a) .   EC50 values were 283, 218

 and 214 ^g/L for chlorophyll a content at 3, 5, and 7 days,  respectively

 (Abou-Waly et al. 1991b) .  Fairchild etal. (1994a,  1998) reported a 4-day
                                ii
 EC50 of 117 /zg/L for chlorophyll content, while Kallgvist and Romstad (1994)

 obtained 3-day growth-based EC50 values of 200 and 110 M9/L-  Photosynthetic

 "C uptake was almost completely inhibited  (99 percent) within 22 hours at an

 exposure of 2,667 /zg/L (Peterson et al . 1994):  A 96-hour EC50 of  147 ^g/L was

 reported by Gaggi et al .   (1995)  for chlorophyll a content.  Additional cell

 number-based EC50 values  reported for 72- to 96-hour exposures include 118.2

 Atg/L (Radetski et al .  1995),  359 ,ug/L (Van der Heever and Grobbelaar 1996),

 200  and 220 M9/L (Abdel-Hamid 19.96), and 26 f^g/L,  (Caux et al . 1996).  Van der

 Heever  and Grobbelaar  (1997,  1998)  expanded on their 1996 study and reported a

 30-minute EC50 value based on decreased oxygen evolution of 222 ^g/L  (1997)

 and  a 4rhour EC50 value based on chlorophyll a fluorescence of 232 M9/L

 (1998) .   Benhra et al .. (1997) .reported an EC50 of 164.3 ^g/L based on growth

 inhibition and Fairchild  et al.  (1997) reported a biomass-based EC50 of 235
      Two  tests  with Stig-eoclonium tenue yielded 24-hour EC50 values based on

"C uptake  of 127 and 224 ^g/L» while a test with Ulothrix subconstricta

yielded an EC50  of only 88 M9/L (Larsen et al .  1986).

      Several  diatom species have been tested for their sensitivities  to

atrazine.   Chlorophyll a content iri the benthic diatom, Craticula  cuspidata,

was significantly reduced after 12 'days exposure to 83 Atg/L atrazine

immediately following 67 days in 1 ^g/L atrazine (Nelson et al .  1999).
                                       30

-------
Cyelotella meneghiniana.  yielded 7-minute  EC50  values  based upon photosynthesis

between  99 and  243 ^g/L  (Millie and Hersh 1987),  while  a  22-hour exposure to

2,667 ^g/L of atrazine inhibited photosynthetic' "c uptake by 97 percent

(Peterson et al.  1994).   A 6-day growth-based  EC50  of 430 pjg/L was obtained

for an unidentified  species of  Cyelotella by Kallgvist  and Romstad (1994).

Hughes  (1986) and Hughes et al.  (1986,  1588) determined several.endpoints in,
                                ii
S-day exposures of Navicula pelliculosa to atrazine,  including a 5-day EC50 of

60 M3/L  based on  cell numbers.   Using a 9-day  recovery  period following the 5-

day exposure, they determined algistatic  and algicidal  concentrations of 1,710

and >3,200 ^g/L,  respectively.   Likewise,  photosynthesis  was  almost completely

inhibited (99 percent) in Nitzschia sp. by a. 22-hour  exposure to 2,667 ^g/L of

atraaine (Peterson et al.  1994).   The cryptomonad,  Cryptomonas pyrinoidifera,

which also appears to be somewhat  less  sensitive to atrazine, had a 6-day EC50

based on growth of 500 ^g/L (Kallqvist  and Romstad  1994).

      The duckweed,  Lenuia minor, when exposed  to 20 /J-g/Ii  of atrazine for 20

days, did not exhibit any adverse  effects,  but reduced  growth occurred at

concentrations  of 50 to  250 ^g/L (Beaumont et  al.  1976,a,b, 1978).  Peterson

et al. (1994),  on the other hand,  observed that growth  was inhibited 95

percent  by a 7-day exposure to  2,667 ^g/L.   Four-day  EC50 values for L. minor

based on biomass  and frond production were 153 and  92 MS/L, respectively

(Pairchild et al. 1997,  1998).   Biochemical and ultrastructural changes in the

chloroplasts of Lemna minor were observed in 15-day exposures of 100 and 1000

^g/L of  atrazine  (Grenier et al.  1979)  as well as an  exposure of 248 Mg/L

(Grenier et al. 1987, 1989; Simard et al. 1990)  for 15,  10 and 2 days,  '

respectively.   This  is very close to the  EC50  of 170  ^g/L for frond production

obtained when Hughes (1986) and Hughes et al.  (1936,  1988) exposed a different

species,  of duckweed, Lemna gibba.,  to atrazine  for 5 days.  Using a 9-day
                                       31

-------
 recovery period,  the  phytostatic and phytocidal concentrations were 1,720 and


 >3,200 Mg/L/ respectively.


       Exposure of wild rice, Zizania aquatics, to  50 ^g/L  of atrazine for 8.3


 days resulted in  a visible  state of senescence and a' 75 percent reduction in


 chlorophyll a in  the  leaves  (Detenbeck et al. 1996).  wild celery,  Vallisneria


 americana,  exhibited  reduced leaf growth and whole plant biomass at an
                                 i

 exposure of 8 pg/L and reduced over-wintering success of tubers at  4 ^g/L


 (Conn 1985).  A 42-day test using this species resulted in an EC50  based on


 total leaf  length of  163 ^g/L  (Davis 1981; Forney  and Davis 1981).   A 14-day


 EC50 based  on wet weight of 22 ^g/L was reported for coontail,  Ceratophy21ua


 sp.  (Fairchild et al.  1998), and reduced stem elongation occurred within 6 to


 8  days at 50 pg/L (Detenbeck et al. 1996).  These  authors  also found that:


 cattails, Typha. latifolia, were unaffected at 25 ^g/L atrazine after 19 days.


 The  Eurasian watermilfoil, Myriophyllum heterophyllwn, had a 14-day wet


 weight-based EC50 of  132 //g/L  (Fairchild et al. 1998) while Myriophyllum
                                                        ..<'-""

 spicatum had a 28-day. EC50 based on length of 1,104 //g/L (Davis 1981;  Forney


 and  Davis 1981) .   This species also exhibited a 30 percent increase in net


 photosynthetic rate at 10 ^g/L after 24 hours (Hoffmann and Winkler 1990), and


 a  50  percent reduction in branch number at 3,700 Aig/L after 5 days  (Bird
                a                      •
 1993) .   Sago pondweed, Potamogreton pectinatus, on  the other hand, had reduced


 biomass  after 28  days at 100 ^g/L (Fleming et al.  1991), and bushy  pondweed,


Najas  sp.,  had a  14-day wet weight-based EC50 of 24 //g/L (Fairchild et al.


 1998).   A 14-day  biomass-based EC50 of'<38 ^g/L was reported for Egeria sp.


 (Fairchild  et al.  1994a).


       The exposure of J-Modea canadensis to atrazine for 21 and 28 days


 resulted in EC50  values based on length of 109 and 80 Atg/L,  respectively


 (Davis 1981; .Forney and Davis 1981).  In a 20-day  exposure to 10 ^zg/L of
                                       32

-------
atrazine,  Hoffmann and Winkler (1990)  found that the dark respiration rate of

this  species exceeded the net photosynthetic rate,  while Detenbeck et al.

(1996)  reported that growth was unaffected after 19 days at 75 ^g/L.

Fairchild  et al.  (1998)  reported a 14-day EC50 of 21 ^g/L for E. canadensis

based upon wet weight.


      Three species of water moss (Fontinalis antipyretics, Fontinalis
                                ,1
hypnoidss  and Fontinalis sguamosa)  were tested by Hoffman and Winkler (1990) .

While F. sguamosa and F. antipyretica  were affected in their photosynthetic

production at 10  M9/L after 24 hours and 20 days, respectively, F. hypnoides

exhibited  a much  greater reduction (90 percent)  in net photosynthesis within

24-hours at an exposure of only 2 (J.g/1..  Conversely, Johnson  (1986) found that

10 Atg/L stimulated growth of mixed macrophytes,  Cera tophyll urn sp. and Elodea

sp. ,  but that 100 and 1,000 Aig/L decreased plant biomass after 30 days.

      The  protozoan, .aca.nt.hainoe.ba castellanii, had population decreases of

from  5  to  40 percent when exposed for  6 days to atrazine at concentrations

from  100 to 10,000 ^9/L (Prescott et al. 1977).   Photosynthesis was inhibited

by about 11 percent in Euglena gracilis at 10 pg/L after 8 hours, and

exhibited  increasingly greater inhibition at higher concentrations  (Valentine

and Bingham 1976) .  Two species of protozoans, Colpidiuzn campylum and

Tetrajjymena pyrifonnis,  had 24-hour EC50 values of >50,000  (Roberts et al.

1990) and  118,500 A/g/L  (Huber et al. 1991), respectively.  Schafer et al.

(1994)  reported a 48-hour EC50 of 96,000 (J.g/1, for T. pyriformis.

      Relatively high concentrations were required to produce notably adverse

responses  in representatives from higher animal phyla.  A concentration of

5,000 pg/L reduced the budding rate in Hydra, viridds after 21 days  (Benson and

Boush 1983) .  The rotifer, Brachionus  calyciflorus, had a 2-4-hour LC50 of

7,840 pg/L (Crisinel et al. 1994).  Two species of leeches,  Glossiphonia
                                       33

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 complanata  and Helobdella stagnalis,  had LC50 values of 6,300 and 9,900 jug/L,


 respectively,  after a 27- to 28-day exposure (Streit and Peter 1978).  After


 21 weeks,  snail (Lymnaea palustris) growth,  fecundity and tissue glycogen


 content /were unaffected-at concentrations up to 125 ^g/L (Baturo et al. 1995),


 but  the activities of benzo[a]pyrene  and glutathione-s-transferase enzymes


 were inhibited at 5 j^g/L (Baturo and  Lagadic 1996).  The 24- and 48-hour LC50

                                 i'
 values were greater than 60,000 ^g/L  for both larval and juvenile mussels,

 Anadonta imbecilis (Johnson et al.  1993) .


       The anostraca'n crustacean, Streptocep-halus texanus,  had a 24-hour LC50


 of >30,000  ^g/L (Crisinel et al. 1994).   The cladoceran, Cez-iodaphnia duJbia,


 exhibited maximum acceptable toxicant concentrations (MATCs) of 7,100 and


 14,100 ^g/L in two 4-day tests  (Oris  et  al.  1991).   A 26-hour LC50 of 3,600


 /^g/L was reported for Daphnia magma (Frear and Boyd 1967).   In 48-hour


 exposures of Daphnia magna  to a nominal  atrazine concentration of 10 ^g/L,


 whole body  residues were only 4.4 and 2.2  times greater than the nominal

• concentration  in  water (Ellgehausen et al.  1980).   Young production was


 reduced in  D.  magna after 21 days.at  2,000 M9/L (Kaushik et al. 1985).  After


 96 hours of exposure,  Bogacka.et al.  (1990)  observed a 30 percent mortality in


 D. magna at 16,900 ^g/L,  and a  60 percent mortality at 48,300 ^g/L.  Johnson


 et al;  (1993)  reported a 48-hour LC50 of 9,400 f^g/l,, but the animals were fed


 at 24 hours.   Crisinel et al. (1994)  obtained a 24-hour KG50 of >30,000 f.tg/1*,


 while Detenbeck et al. (1996) observed a significant decrease in the survival

 of these invertebrates after 48 hours of exposure at 25 ^g/L, but not at 50


 pg/L.  Nishiuchi  and Hashimoto  ,(1967/ 1969)  found the 3-hour LC50 to be


 greater than 40,000 £/g/L for Uaphnia  pulex.   Exposures of D. pulex for 28 to


 approximately  70  days resulted  in decreased, survival and reproduction at

 concentrations ranging from 1,000 and 20,000 ^ig/L atrazine,. with reproduction
                                        34

-------
 affected more than survival (Schober ana Lampert 1977) .   Food consumption was




 reduced by 10 percent at 350 jug/L and by 5o'percent at 1,600 ^g/L after 10




 minutes .(Pott 1980).  Bowman, et al.  (1981)  reported an 18-hour LC50 for D.




 pulex of approximately 700 /zg/L.  Conversely,  the 3-hour LC50 was in excess of




 40,0,00 ^g/L for the cladoceran, Moina macrocopa (Nishiuchi and Hashimoto 1967,




 1969),  and a concentration of 1,000  ^g/L was shown to cause 40 percent




 mortality and reduced populatio'n growth after 4 to 6 weeks (Shcherban



 1972a,b).





       The  amphipod,  Gammarus fasciatus,  had a 48-hour LC50 of 5,700 ^g/L




 (Macek et  al.  1976) .  Similarly, exposure of HyaJella azteca for 18 hours




 resulted in an LC50  of 2,000 ^g/L (Bowman et al. 1981).   For the midge,




 Chironomus riparius, a. 10-day exposure to atrazine yielded an LC50 of 18,900




Atg/L  (Taylor et al.  1991),  while a 96-hour exposure of C. tentans in a fed




 test  had less  than 50 percent mortality at the high concentration of 28,000




Pg/L  (McNamara 1991b) .  The 18-hour  LC50 for the white dotted mosquito, Culex




restuans,  is considerably higher at  approximately 60,000 ^g/L (Bowman et al.



1981).





      Rainbow  trout, .Oncoriynchus myklss,  embryos and sac fry exposed




continuously for^ 23  (embryos at hatching)  and 27 (sac fry, 4 days post-hatch)




days  had LC50  values between 696 and 888 ^g/L (Birge et al. 1979).  Water




hardness did not have any appreciable effect.   A. concentration of 4,020 ^g/L




was required to produce over 60 percent teratic larvae.   Pluta  (1989) reported




a 48-hour  LC50 of 5,660 ^g/L-   Changes" in the ultrastructure of trout renal




corpuscles and tubules were observed following 28-day exposures to 5 to 10




Mg/L  of  atrazine (Fischer-Scherl et  al.  1991).  Similarly, 28-day exposures




resulted in slight ultrastructural changes in trout renal corpuscles at 5




    / slight histopathological changes in the liver and increased
                                       35

-------
 ultras true tural changes  in  renal corpuscles at 10 ^g/L,',  and in further  changes

 in renal corpuscles  and  liver  cells at 20 ptg/L (Schwaiger  et al.  1991).  A 14-

 day exposure to 10 ^g/L  of  atrazine did not affect survival,  body weight,

 liver weight or liver  enzyme activity  (Egaas et al. 1993) .   Exposure to


 concentrations of 3.0  and 50 ^g/L for 10 days were reported to reduce plasma

 protein in rainbow trout (Davies et al. 1994b) .  Oulmi  et  al.  (1995)  observed
                                 ,i
 kidney changes at the  cellular level within 5 weeks in  O. mykiss  in the

 proximal tubules, at  12.4 pg/L,  and in both the proximal and distal tubules at

 24.0 //g/L.  The most sensitive indicator of biological  effect exhibited by a

 salmonid was the reduction  in  milt and plasma levels of 17,20 |3-dihydroxy-4-

 pregnen-3-one in Atlantic salmon, Saline salajr, parr exposed for 5 days  to,0.04

 Mg/L atrazine (Moore and Waring 1998) .  In the same study,  a 30-minute

 exposure to 2.0 /^g/L reduced olfactory response of mature  male parr to  a

 female pheromone.


       The 48-hour LC50 for  the goldfish, Carassius auratus,  was >10,000 ^g/L

 (Nishiuchi and Hashimoto 1967,  1969), although Sag"liq and  Trijasse (1998)

 observed reduced burst swimming performance in goldfish after a 24-hour

'exposure to 0.5 Aig/L.  The  48-hour LC50 for the common  carp,  Cyprd.nus carpio,

 was  also >10,000 Mg/L  (Nishiuchi and Hashimoto 1967, 1969)'.   Short-term

 exposures of from 4  to 24 hours to lesser concentrations between 100 and  500

 Mg/L resulted in increased  serum cortisol and serum glucose (Hanke et al.

 1983.) .  Serum acetylcholinesterase first increased and  then decreased with

 time of exposure.  Changes  were also noted in gill ATPase  activity.  Longer

 exposures- of 72-hour duration  to 1,000 yag/L and 100 ptg/L of atrazine also

 yielded decreased liver  glycogen  (Hanke et al. 1983), and decreased liver and
             ' !                                            "
 muscle glycogen as well  as  serum protein, and cholesterol (Gluth and Hanke

 1984,  1985), respectively.  Juvenile carp yielded a 48-hour LC50 of 16,100

-------
      (Pluta 1589),  and a 96-hour LC50,  in  which the fish were fed,  of 18,800



      (Neskovic  et al.  1993).   It was  noted in the latter study that



biochemical changes in the serum,  heart, liver and kidneys of carp were



observed after  14 days of exposure to 1,500 /ig/L,  as well as hyperplasia of



gill  epithelial cells  (Neskovic et al.  1993).   Conversely, no effects on gill,



liver,  and histopathology were observed at this same concentration (1,500

                                t'
/^g/L)  in a study by Poleksic  et al.  (1997) .



       Jop (1991c)  reported the "no observed effect concentration" (NOEC)  to  be



in excess of 4,900  ^g/L for fathead minnows,  P. promelas,  exposed to atrazine



for 7  days.   Also,  survival and growth were shown to be unaffected in fathead



minnows  exposed to  75  Mg/L for 13  days (Detenbeck et al. 1996) .   On the other



hand,  channel catfish  (Ictalurus punctatus)  embryos and sac fry had LC50



values between  176  and 272 jug/L after exposures of either 4.5 (embryos at



hatch) or 8.5 (sac  fry,  4 days post-hatch)  days' (Birge et al. 1979).



Concentrations  of approximately 340 ^g/L caused an incidence of 13 to 16



percent  teratic larvae,  while concentrations of approximately 3,850 Mg/L


resulted in 47  to 69 percent  teratic  larvae.



      Mosguitofish  (GamJbusia  affinis)  survival was unaffected in a 48-hour



exposure to 10,000  ;/g/L (Darwazeh and Mull a 1974),  and LC50 values as high as



38,200 and 31,600 ^g/L were reported  for the guppy (Poecilia reticulata)  after



exposures of 48 and 72 hours,  respectively (Tscheu-Schluter 1976).  These data



are consistent  with results reported  by Bogacka et al.  (1990), in which the



authors  reported mortalities  of 40 and"53.2 percent after exposing guppies for


96 hours to 28,600  and 37,200 ^g/L, respectively.



       Exposure  of the  Mozambique tilapia,  Tilapia mossambica, to 1,100 ng/I* of



atrazine for 30 to  90  days affected blood  composition, oxygen consumption,



water  content,  and  the biochemistry of the brain and liver  (Prasad et al.
                                       37

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 1991..b, Srinivas et al. 1991).* A  90.day exposure also resulted in increased




 serum sodium and potassium, and decreased serum calcium', magnesium and




 bicarbonate (Prasad and Reddy 1994).





        The embryo and larval stages  of several amphibian species  were'exposed




 to atrazine (Birge et al. 1980), the results of which are quite  different




 between  species (Table 6).  LC50 values for continuous exposure  of embryos and




 larvae through 4 days post-hatch were 410 pg/L for the bullfrog  (Rana




 cate^eiaaa),  7,680 ^g/L .for the leopard frog (Sana pipiens) .  17,960 ^g/L for




 the pickerel  frog (Ran* palustris)/ and >48,000 pg/L for the American toad




 (Bufo americanus) .   In most of these species,  concentrations of  atrazine in  -




 excess of '5,000 ^g/L were required to cause an incidence of  teratic'larvae in




 excess of 7 percent.   Survival and growth of *.  pip±ens tadpoles were




 unaffected after 41  days of exposure to 25 Mg/L (Detenbeck et  al.  1996).  A




 95-hour exposure of  the African clawed frog Ueoopua laevia) embryos to 8,000




 ^g/L  resulted in 100  percent abnormal embryos  (Morgan et al. 1996).  The




 lowest observed effect  concentration (LOEC;  teratogenesis)' in"the  study was




 1,100 ^g/L.  This concentration is  more than an order of magnitude higher than




 that  which delayed development and  retarded the growth in the  tiger




 salamander, Ambystoma  tlgrivm-,  after 86  days of  exposure (Larson et al.  1998).




       Several aquatic ecosystem studies,  either  artificial laboratory-




microcosms or field mesocosms,  have provided valuable insight  into ecosystem




structural and  functional responses to  atrazine.   A mixed assemblage of algal




species exposed  to 10 ^zg/L  of  atrazine-for periods of time ranging from 1 day




to 3  weeks exhibited reductions  in  gross  productivity between  39 and 78    '




perc-ent  (Kosinski and Merkle 1984;  Kosinski  et  al.  1985).   Exposure of an




experimental stream periphyton community  to  1,,000  pg/L for 14  days caused




severe population density reductions in  several  species,  and total destruction
                                        38

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of the green  alga,  Cladophora grlomerata  (Kosinski  1984).  The extreme toxicity

to C. gloaerata  is  notable because of  the  dominant role that it often plays  in

structuring a benthic community.   Similarly,  Moorhead and Kosinski  (1986)

observed  reduced net primary productivity  at  100 #g/L in an assemblage of

mixed stream  algal  species.  By contrast,  in  a mixed stream community, no

effects were  observed upon stream aacroinvertebrate community structure,
                                ,i
periphyton production or biomass,  or the community photosynthesis/respiration

ratio following  a 30-day exposure-at 25 jUg/L  (Lynch et al.  1985).

      Malanchuk  and Kollig (1985)  observed chemical changes in an experimental

stream community consisting of microscopic autotrophs and heterotrophs

following the introduction of atrazine at  a nominal concentration of 100 ^tg/L

for a 2-week  exposure period, after which  time the atrazine was removed from

the ecosystem.  They observed decreased diurnal fluctuations In pE  and

dissolved oxygen concentrations,  as well as lower  mean values for these

parameters while atrasine treatment was on-going,  but nitrate nitrogen levels

increased.  Following the cessation .of atrazine treatment,  there was a rapid

recovery  for  each of these parameters  back to control levels.

      Biomass reductions were also noted in a stream aufwuchs community •

exposed to 24 or 134 ^g/L of atrazine  for  12  days   (Krieger et al. 1988),

although  a 24-hour exposure of 77.5 ^gfL had  no effect upon algal cell numbers

or biomass in a  natural stream periphyton  community  (Jurgenson and  Hoagland

1990).  An exposure of as low as 0.5 (J.g/1*  for 6 months resulted  in  an initial

decrease  in phytoplankton species followed by a recovery (Lakshminarayana et

al. 1992).  Gruessner and Watzin  (1996), however,  did not.observe any effects

of atrazine on a stream community of  a'ttached algae  and benthic  invertebrates

at a concentration of 5 Aig/L when exposed for 14 days.  Pearson  and Crossland
                                       39

-------
 (1996)  reported  an  inhibition of photosynthesis by  the  periphyton community of




 an artificial stream  following exposure of 100 Mg/-L of  atrazine for 3KD days.




       In a static pond microcosm (1 L beaker), Brockway et al.  (1984) found




 that a 7-day exposure to  5.0 ,/g/L had no effect upon diurnal oxygen




 production, a measure-of  photosynthesis, by the various species of green and




 blue-green algae present.  A 50 ^g/L exposure for 12  days  resulted in a 25 to




 30  percent reduction  in diurna}' 'oxygen production,  while 7- to  12-day,




 exposures at 100 to 5,000 pg/L further decreased oxygen production. Berard et




 al.  (1999)  observed seasonal and species-dependent  effects in a lake microcosm




plankton community after  10 to 21 days of exposure  to 10 Mg/L atrazine.




During  the experiment,; growth was generally stimulated  for Chryptophytes and




Chrysophytes,  but inhibited in Chlorella vul&arls.




      Exposure of a freshwater microcosm to 5.1 ^g/L  of atrazine for 7 weeks




did  not  affect the-species composition of phytoplankton, zooplankton or




benthic  macroinvertebrates, but did cause a slight  decrease in  photosyathetic




activity (Van den Brink et al. 1995).   Hamala and Kollig (1985) found an




approximate 75 percent decrease in the productivity/respiration (P/R) ratio in




a 14-day exposure to  100 W/L of a periphyton-dominated microcosm which




contained 33  algal taxa.  They also observed reduced  algal densities,




decreased species diversity,  altered species composition and reduced biociass




accumulation.   In a 21-day recovery, period, net community  productivity




returned to control values within IS days, while very little recovery occurred




in community structural parameters.  This fairly rapid  recovery in a




functional  parameter  indicated that the primary effect  of  atrazine at, this




exposure level 'was algistatic and not algicidal for those  species involved in



the  recovery,.                    •                                ,
                                       40

-------
       Stay et al.  (1985), using a 3.7 L laboratory microcosm  consisting of 10


algal  species and 5 animal species (one protozoan, one rotifer,  and  three


crustaceans), found that reduction in the ratio of »c  uptake/chlorophyll a was


Che most sensitive measure of atrazine effect.  This suggested  that  the


effectiveness of the photosynthetic system was impaired.  The lowest exposure


(i.e.,  43.8 pg/L over 60 days) resulted in significant reductions


(approximately 60 to 90 percent')' in the ratio throughout most of the study.


Higher exposures (nominal concentrations of 100 to 500 /zg/L)  caused  further


reductions in this ratio, but not as large a difference as between controls


and the lowest exposure.


       Peichl et al. (1984)  observed changes in the population densities of


sooplankton in a pond mesocosm study after 70 days of exposure  to 200 ^g/L' of


atrasine.   In a later study (Peichl et al. 1985), the authors observed changes


in the  phytoplankton community after 121 days- of exposure to  only 10 ^g/L.


Experimental ponds in Kansas that were exposed for several years to  single


annual  applications of atrazine at nominal concentrations of  20  Mg/L or-more


exhibited reductions in the production and biomass of phytoplankton, in


macrophyte populations and in populations of benthic insect grazers, bullfrog


(R. catesbeiana)  tadpoles,  grass carp (Ctenopharyng-odon idella)  that had been


introduced,  and in bluegills  (deNoyelles et al. 1982, 1989, 1994).   Initi.il


nominal concentrations- of 20,  100, 200 and 500 Mg/L depressed phytoplankton


growth within a few days in the ponds.  However, after 3 weeks,  phytoplankton


production and biomass were similar to'controls.  deNoyelles  and Kettle  (1985)


observed reduced photosynthesis of 40 percent or more  in short-term  (24-hour)
                       ft

bioassays at these same atrazine concentrations, but longer-term bioassays  (20


days)  and the experimental pond studies showed a recovery from  this  initial


reduction.
                                       41

-------
      Benthic insect community structure was studied in the same experimental




ponds used in'Kansas following two single annual treatments at 20, 100 and 500




/ig/L  (Dewey 1986;  Dewey and deNoyelles 1994) .   Significant reductions of both




species  richness and total abundance 'of emerging insects was observed at the




lowest exposure of 20 ^g/L.  Abundance of the herbivorous, non-predatory




insects  was reduced at 20 ^9/L, but not abundances of the predatory species.




This  indicated that the observed loss of total insects was a secondary effect




due to feeding habit and loss of plant life, rather than a direct  toxic




effect.   Loss:of insect habitat, particularly in the form of macrophytes, also




likely had some effect upon the insect community.  These effects tended to




destabilise the ecosystem  (Dewey and deNoyelles 1994) .




      Species composition of macrophytes was altered in a pond mesocosm




community following an 8-week, exposure to 50 ^g/L of •atrazine  (Fairchild et




al. 1994a).   However, functional parameters were unaffected, indicating




functioning redundancy within the ecosystem.  Juttner et al.  (1995) did not




observe  any effects upon the plankton community of a pond mesocosm following a




2-month  exposure to 5 ng/Z,, but did observe decreased oxygen production, pH




and conductivity at 10 ^g/L, and decreased phytoplankton populations  at 182




Mg/L-.  At 318 ^9/L/ reproduction was affected in DapJmia loug-ispina  and a




population of rcfcifers, Polyar-tira sp., was eliminated.




      In a laboratory microcosm using a naturally derived microorganism




community,  Pratt et al.  (1988) observed that a. 21-day exposure  to  a mean




measured concentration of  10 Aig/L of atrazine did not affect  the dissolved




oxygen,  a measure of photosynthetic function, but that  a  concentration of  32.0




^g/L  caused significant reductions in this  parameter.   This resulted in a




calculated maximum acceptable  toxicant concentration (MATC)  of 17.9 jug/.t based




upon  this 'functional endpoint.  Several other  endpoints,  such as protozoan
                                        42

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colonization, biomass protein,  chlorophyll a and potassium levels, were less




sensitive than dissolved oxygen,  and had a calculated MATC of 193 W/l,.




       Stay et al.  (1989) studied atrazine effects in 1 L laboratory microcosms




containing mixed phyto- and zooplankton cultured from three Oregon lakes and




one  pond.  A 42-day exposure of approximately IS W/L atrazine did not affect




net  primary productivity, the P/R ratio, or pH,  but these parameters were




significantly reduced fr.om controls at a mean measured concentration of




approximately 84 pg/L.





       Larsen et al.  (1986)  measured photosynthetic "C uptake in  a 3 L Taub




microcosm community at different time intervals for up to 373 days after




treatment with atrazine.  EC50  values ranged from 24 jug/L at 177  days to 131




//g/L at  43 days after atrazine  treatment.




       A  50 nr pond community exposed to  atrazine for 4 months at a




concentration between 60 and 120 ^g/L eliminated a population of  duckweed,




Lemna  minor,' within 27 days (Gunkel 1983) .  Gunk'el also observed  a rapid




succession of algal species and a reduced rate of reproduction in Daphnia




pulicar-ia.  Treatments of a pond mesocosm community for 2 years with 20, 100




and  300  ^g/L of atrazine caused decreases in cell numbers of green algae and




of cladoceran populations,  but increased numbers of cryptomonads  (Neugebauer



et al. 1990).





       In experimental ponds treated in May and June with 20 ^g/L  of atrazine




for  two  years,  there was decreased abundance of Endochironomus nigricans in




June and of total macroinvertebrates in' both May and June, followed by




recovery in July (Huggins et al. 1994).  Epiphytes, detritovores  and  '




generaliats also, exhibited initial decreases in populations, followed by a



recovery.
                                        43

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      A  short-term exposure (>3 hour)  of pond algae to 10 i^g/'L  of atrazine was




observed to increase the rate of fluorescence for photosystem II  (Ruth 1996).




      In two reports of studies conducted at the same site, a lake community




was .enclosed with a limnocorral (5mx5mx5m deep) to which atrazine was




added.   Both studies focused on the periphyton community.  In the first study




(Herman  et al.  1986), the limnocorrals received two nominal atrazine




applications of 100 .^g/L, one oh day 0 and another on day 35.   After 34 days




of exposur.e to  measured concentrations ranging between 80 and 140 ^g/L, a




reduction in periphyton ash-free dry weight was observed.  Over a 9-week




period with two atrazine applications 6 weeks apart, which resulted in




measured concentrations of approximately 80 to 140 £ig/L after the first




application and 110 to 190 ^g/L after the second application, reductions




occurred in chlorophyll a,' organic matter and total periphyton  algal bioinass.




In the second .study  (Hamilton et al. 1987), a 230-day exposure  to a mean




measured atrazine concentration of 80 ,ug/L caused approximate reductions of 50




percent  in biomass, 22 percent in cell numbers and 32 percent in  number of




species.   The results were more pronounced in exposures to mean measured




atrazine concentrations of 140 and 1,560 ^g/L.  A shift in community  structure.




occurred from a chlorophyte-dominated community to a diatom-dominated




c omnvuni ty.





      AqTiatic enclosures .exposed to a nominal atrazine application  of 100  /J.g/'L




on June  1 followed by a second application of the same concentration  35 days




later, exhibited a gradual die-off of the phytopiankton,  a  long period of




recovery for the green algal community, and a distinct shift in the taxonomic




composition of algae  (Hamilton et al. 1988) .  Thirteen days  after the first




application, significant declines occurred  in populations of the green algal




species  Elaka.tothrix gelatinosa, Tetraedon minimum,  Sphaerocystis schroeteri,
                                        44

-------
 and Oocystis lacustris, and of the dinoflagellate. Gynwodinlum spp.  Seventy-




 seven days after the second application, phytoplankton communities were still




 distinctly different, and total fresh weight biomass was  reduced.  By 323 days




 after the first application, the phytoplankton assemblages  were again similar




 between control and treated enclosures.  From day 1 to day 114,  control




 enclosures had an average of five more taxa than the atrazine- treated




 enclosures.   During the period Between days 49 and 77, the  green algal




 (Cfalorophyta)  biomass represented <7 percent of that found  in  the controls.




 By  the following spring (day 323), the biomass had returned to  control levels.




 The  herbicide treatment did not affect the rotifer or crustacean communities.




 In  the same  exposures, Hamilton et al. (1989)  observed that the atrazine-




 treated enclosures became clearer with increased Secchi disc readings, while




 readings of  dissolved oxygen,  chlorophyll, dissolved organic carbon, and




 particulate  organic carbon decreased.




       Using  1.70 m'  enclosures  in  a  moderately eutrophic lake,   Lampert et al.




 (1S89)  observed decreased photosynthesis and decreased populations of certain




 zooplankters at atrazine concentrations of 0.1 and 1.0 jug/L.   At 0.1 ^g/L,




populations  of Dapimia sp. were severely reduced within 15  days, and oxygen




 concentrations were reduced after 10 days.  At 1.0 ^g/L,  concentrations of




 chlorophyll  a aj?d oxygen were reduced after 18 days as were populations of




Dap&nia,  Cyclops, and Bosmina species, and nauplii larvae.   At 0.1 ^g/L,  there




was  an apparent recovery after about 25 days.   Genoni  (1992) observed a




 decreased algal population density and'a decreased "scope for  change in




 ascendency"  in a microcosm community exposed to 250 Aig/L.   The scope for




 change in ascendency is a biological system response endpoint,  considered to




be analagous to the scope for growth endpoint for individual . organisms.
                                       45

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       Gustavson and Wangberg  (1995)  observed some minor changes  in species




composition of the phytoplankton community in a lake mesocosm community after




a  20-day exposure to  20 Mg/L.  ECSO  values we're 58 and 52 ^g/L for the




phytoplankton community, and 52 and  5.4 M9/L for the periphyton community.




Brown and 'Lean (1995) :found that a short-term exposure (3 hours)  of lake




phytoplankton to atrazine resulted in a much lower ECSO based upon




photosynthetic carbon assimilation (.i.e.,  100 ^g/L), than when based upon




phosphate or ammonium assimilation (14,000 and >33,000 ^g/L,  respectively).  A




stream periphyton, community exhibited a significant reduction in chlorophyll a




following a brief exposure (<4 hours)  to 109 Mg/L of atrazine (Day 1993).




Caux  and Kent (1995)  observed a reduction in green algae in Quebec streams




following the spring  atrazine runoff pulse,  with a maximum stream




concentration of approximately 40 ^g/L.  'Detenbeck et al.  (1996) 'observed a




decrease in the gross productivity of a wetland mesocosm community'after 9'to




27 days  of exposure at-an atrazine concentration of 15 ^g/L-   There also was




an increase in the concentrations of dissolved nutrients in the  water.




       From the various studies of ecosystem effects (i.e., microcosm, mesoccsin




and limnocorral studies), the lowest concentrations of atrazine  that'have,




resulted in temporary negative effects upon abundance of aquatic plants




(primary effect)  and  animals (secondary effect)  have generally occurred at 15-




20 ^g/L  and above.   Studies, by Berard et al. '(1999), Kosinski and Merkle




(1984),  Kosinski et al. (1985), Lakshminarayana et al. (1992),  Lampert et al.




(1989),  and Peichl et al. (1984, 1985);"have observed effects  at  lower




concentrations.   It appears that for effects at concentrations up to 15 /;g/L,




the communities can recover quite rapidly following dissipation of the




atrazine concentration.  In a review of. microcosm and mesocosm studies with




atrazine,  Giddings and Biever  (1994)  concluded that concentrations of 20
                                       46

-------
or less  typically caused minor effects,  if  any,  on primary production and


plant community composition,  and recovery occurred quickly, even if atrazi'ne


remained in the system.  Concentrations  above 50 ^g/L caused more severe


reductions  in productivity, plant biomass, ^and community structure, as well as


indirect effects on herbivorous invertebrates and fish.  They further stated


that freshwater ecosystems can recover from the effects of atrazine in a year

                                ii
or less  except at very high exposures of 500 ng/I. or greater.


      In summary, cyanobacteria had EC50 values for various exposure durations


of 30 pg/L  or greater, while EC50 values for green algae, diatoms and


cryptomonads were £15 ^g/L.  Among macrophytes,  duckweed had a minimal 4-day


EC50 of  92  /ig/L.  Wild rice was affected at 50 ^g/L/ and wild celery had


reduced  growth at 8 /ng/L.  Several rooted vascular plants  (i.e., coontail,


bushy pondweed, egeria, and elodea) had 14-day EC50 values between  21 and <38


A*g/L, while that for a water milfoil was 132 >g/L.  Two species  of  water moss


(•FontiBalis sp.) exhibited reduced photosynthetic activity at 10 ng/l, and one


species  was affected at 2 ^g/L.  EC50/LC50  values for protozoans,


coelenterates, annelids, molluscs and rotifers were £S,3PO >g/L.  Various


crustaceans had LC50 values £5,700 ^g/L-  The most sensitive endpoints among


fish were rainbow trout plasma protein and kidney ultrastructural  changes at


atrazine exposures of 3 and 3.5 fJ.g/'L, respectively.  The lowest  LC50  values -is


fish were 176-272 pg/L for 4.5 to 8.5-day exposures with early  life-stages of


channel  catfish.  Frog embryo and tadpole life-stages had  LC50 values £410


Atg/L.  In most aquatic ecosystem studi-es,  reductions in algal  or vascular


plant bicmass were observed at concentrations. £15 MS/L.  'This  commonly


resulted in the  reduction  of herbivore populations, as well.   One exception


reported effects at much lower concentrations  (as low  as  0.1 pg/L) .  From


these  freshwater Other Data, most  of  the effect  levels of  possible biological
                                        47

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 significance  appear to be *15 W/L.   This  concentration is greater  than the




 freshwater Final  Chronic Value,  and therefore does not determine  the Criterion



 Continuous Concentration.





       Additional  data are available for saltwater algae, kelp, .submerged




 vascular plants,  emergent vascular plants,  and aquatic animals '(Table 6).




 ECSO values based on differing endpoints  (e.g.,  oxygen evolution  or growth)




 for various green algal species /ranged  from 37 ^g/L to 600 ^g/L  (Gaggi et al.




 1995;  Hollister and Walsh 1973;  Hughes  1986;  Hughes et.al. 1986,  1988,'Samson




 and Popovic 1988;  Walsh 1972).   A 48-hour  exposure of.the green alga,




 Cunaliella bioculata,  to 216  Mg/L of  atrazine resulted in a growth reduction




 of  approximately  35  percent  (Felix et al.  1988).   Seven-day growth tests with




 the green alga, Nannochloris  oculata, at concentrations of SO: and 100 /zg/L




 suggested that atrazine toxicity was  dependent on light and temperature




 (Karlander et al.  1983;  Mayasich,et al. 1986),  although the effect was not




 dramatic.   A concentration of 15 /zg/L changed the doubling time in ff. oculata.



 (Mayasich et al.  1987} .





       Diatom species were similar to  green  algae  in terms of their




 sensitivities to  atrazine.  ECSO values for exposures  of various  durations,




were generally between  20 and 460  Mg/L  (Hollister and  Walsh ~1973; Walsh 1972;




Walsh  et  al.  1988).  Plumley  and Davis  (1980)  observed reduced photosynthesis




 in N±tzsch±a. sigma and  reduced chlorophyll  in Zfaalassiosira fluviatilis in 7-




day exposures; to  220'^g/L.  Mayasich  et al.  (1987)  reported a limited effect




on doubling time  to'pfaaeodactylum tricornutum in  a 7-day exposure to 50 ^g/L



of atrazine.   '





       The  red alga, Porphyridium cruentum,  had an ECSO based on oxygen




evolution  of 79 ^g/L when exposed for 90 minutes  (Hollister and Walsh 1973),




and the kelp/ Laniinajria iyperborea, had a 24-hour LOEC value for  respiration
                                        48

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of >1,000  A
-------
the brown shrimp, Penaeus aztecus,  had a 48-hour EC50 of 1, 000 ^g/L (Butler




1964;  Mayer 1987).  Adult fiddler crabs,  Uca pugna*,  were not very sensitive




to one-time applications of atrazine either in field or laboratory exposures




(Plumley'et al.  1980).  However,  there was a.seasonal effect on the




sensitivity of this species even when the laboratory conditions were the same.




Animals  collected in the summer were more sensitive to atrazine than those




collected in either the' spring or fall.   Two other species of crabs, Sesarma




cinereum and' Pajiopeus sp.,  were also insensitive to very high levels of




atrazine (Plumley et al. 1980).





       The acute  and chronic effects of atrazine on an estuarine microbial




community were recently examined by DeLorenzo et al.  (1999a,b) .  Exposure for




9 days to 40 ^g/L of atrazine in dilute seawater (7-25 g/kg) inhibited the




phototropbic component - chlorophyll a,  carbon assimilation, bio volume, and




caused changes in species composition (DeLorenzo et al. 1999a).  The same




effects  were observed in full strength seawater at an atrazine concentration




of 47 ^g/L,  but  within 24 hours (DeLorenzo et al.  1999b).  An earlier mesocosa




study  of a mixed assemblage of marine phytoplankton species demonstrated




reduced  photosynthesis and primary production at concentrations of 0.12 and




0.56 ptg/L atrazine at 15 days (Bester et al. 1995).  This is much lower than




effe.ct levels demonstrated from the other studies.









Unused Data










      Data from some studies were not used in this document,  as they did  not




meet the criteria for inclusion as specified in the Guidelines  (Stephan et al.




1985).   The reader is referred to the Guidelines for further  information




regarding these  criteria.
                                       50

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       Some data on the effects of atrazine on aquatic organisms were not used

because the studies were conducted with species that are not resident in North

America (e.g., Alazemi et al. 1996; Biagianti-Risbourg and Bastide  1995; Diaz

et  al.  1998; Forget et al. 1998; Gorge, and Nagel 1990; Gunkel and Kausch 1976;

G=hetotskii et al. 1977; Hussein et al. 1996; Juhnke and Luedemann  1978; Kirby

et  al.  1998; Lewis et al. 1993; L'Haridon et al. 1993; Nagel 1992;  Pantani et
                                 i
al.  1997;  Portmann 1972; Prasad>'et al. 1990, 1995; Steinberg et al. 1995).

Results were not used if the duration of the exposure was not specified or was

unclear (e.g., Hopkins and Kain 1968; Portmann 1972; Rojickova-Padrtova and.

Marsalek 1999; Tellenbach et al. 1983), or if the procedures or test materials

were not adequately described or translated  (e.g. Braginskii and Migal 1973;

Delistraty 1999; Xross et al. 1992; Shcherban 1973; Tang et al. 1998a,b;

Wen=el  et  all 1997) .

      The  acute toxicity data of Bathe et.al. (1973, 1975) were not used due

to  as. insufficient number of test organisms.  High control moralities occurred

in  tests reported by Dodson et al. (1999), as well as in a chronic  study with

Gaffiinajrus jfasciatus (Macek et al. 1976) .  Studies published only as  abstracts

of presentations were not used (e.g., Fairchild et al., 1994b; Palmstrom and

Krieger 1983; Zora and Paladino 1986).  Secondary observations reported in a

review  were not used  (e'.g., Giddings and Hall 1998; Eurlbert 1975;  Eutchinson

et  al.  199'8; Lange et al. 1998; Mercuric 1998).  Similarly, papers .by Birge et

al.  (1983), Fairchild et al. (Manuscript), Mark and Solbe  (1998), and Pratt et

al.  (1993, 1997) were not used, as the/data  they contained'had been previously

published.  A study by Butler et al.  (1975) was not used since data from

several algal taxa were grouped in the reporting of results.  Stratton  and

Giles  (1990) expressed toxicity on the basis of cell numbers.
                                       . 51

-------
                                     was
       Toxicity  data from laboratory tests  were generally, not used if  the test




 material was, a  formulation or emulsifiable concentrate  and. atrazine comprised




 less than 80  percent of its weight (e .g. ,  Antychowicz. et.  al. 1979; Carder and




 Hoagland 1998;  Clements et al.  1997; Hartman and Martin 1985;  Hiltibran 1967;




 Kowe et al.  1998;  Lin et al. 1999; Pavlov  1976;  Roj ickova-Padrtova and




 Marsalek 1999;  Semov and losifov 1973;  Sreenivas and Rana 1991,  1994; Walker




 19S4),  if atrazine was a component of .a pesticide mixture (e.g.,  Britson and




 Threlkeld 1998;  Grain et al. 1998; Guasch  and Sabater 1998;  Guasch et al.




 1997,  1998; Hartgers et al.  1998;  Lcwcock  et al. 1997;  Ort et  al. 1994-.




 Pollehne et al.  1999;  Reeder et al.  1998;  Vanderpoor ten 1999)-,  if atrazine




 dosed  in the .diet  (e.g.,  Cossarini-Dunier  et al. 1988), or if  the




 concentration of solvent used in atrazine  stock  preparation exceeded 0.5 ml/L




 (e.g.,  Cheney et al.  1997;  Grain et  al. 1997,  1999; Tang  et al,.  1997) ; the  '




 latter  representing a value  below which neither  acetone nor ethanol are toxic




 to  algae (e.g.. El  Jay 1996; Stratton and  Corke  198.1) ,  but where DMSO and




 atrazine may interact additively (SI Jay 1996) .




      The results  from Langan and Hoagland (1996)  were  not used because the




 tests were conducted in distilled water without  addition  of the appropriate




 salts.  . Toxicity tests by Schmitz  et al.  (1994)  and Tubbing et  al. (1993)  were




not  used because the tests were performed  in river water  which  was likely




contaminated with various'other chemicals.   Similarly,  a  cytopathological




study of fish exposed to  a  spill of  atrazine plus other pesticides was not




used (e.g.,  Spazier et al.  1992).  Effects data  were not  used  if  the atrazine




exposure was .part of a soil  mixture  (e.g.,  Johnson et al.  1999, Jones and




Estes 1984;  Miller  and Doxtader 1995; Ruth 1997).  McBride and Richards (1975)




exposed excised tissue, and  Petit  et al.  (1997)  exposed cell cultures.
52

-------
       A study of atrazine  accumulation by Bohm and Muller (1976) was not used




due to expression of results on a volume basis rather  than a weight basis.  A




bioconcentration study by  Walsh and Ribelin  (1973) was  not used due to the use




of  nominal atrazine  concentrations in the exposure water rather than measured




concentrations.  Data were not used if the exposure was to radiolabel.ed




atrasine (e.g., Davis et al. 1979; Jones et al. 1982; McEnerney and Davis




1979;  Neumann et al.  1987; Pillai et al. 1977, 1979,; Weete et al. 1980),  or




atrazine was not detected  in tissue (e.g., Harris et al.  1998).  Uptake and •




accumulation from exposures in flasks or microcosms were not used if "C only




was measured and not  atrazine itself (e.g., Huckins et  al. 198S; Isensee 1975,




1987;  Kearney et al.  1977; Mailhot 1987).      •




       Biochemical studies  of resistant strains of mutated algae  (e.g., Bousra-




Halfon et al. 1997; Forster et al. 1997; Ottmeier et al.  1991) and results




frcxa in vitro genotoxicity and mutagenicity tests  (e.g.,  Ruiz and Marzin 1997)




were not used.  A study of atrazine effects upon promutagen activation by




Selenastr-uni capricornuturn  (e.g., Sauser and Klaine 1990)  and alteration is.  •




allele and genotype frequencies of the'oyster, Crassostrea g-ic-as (e.g., Moraga




and Tanguy 2000) were also not used.
Summary
      Atrazine is not highly  toxic to aquatic animals on an acute basis.




SMAVs for eight freshwater  invertebrate species  ranged from 720 /J.g/1* for a




midge,  Chironomus tentans,  to 49,000 for'Daphnia magna.   SMAVs for nine fish




species ranged from  5,300 ^g/L for the rainbow  trout, OncorJiynciius mykiss, to




60,000  Aig/L for the  goldfish,  Carassius .auratus  (Figure 1).  Genus Mean Acute




Values  for atrazine  are  available for nine genera  of saltwater animals and
                                        53

-------
 range from 2,324 to >30,000 pg/L; a factor of approximately  12.9  (Figure 2).




 Genus Mean Acute values  for the  four most sensitive genera  (three  epeciea of




 crustaceans and one fish) differed by a factor of approximately 2.5.




       Chronic effects of atrazine exposure to aquatic animals  have been




 studied with seven freshwater species, three of which are invertebrates and




 four of which are fish  (Figure 3).  In three tests with Cerfodaphnia'duMa,




 chronic values were 3,536, 3,536, and 1,732 Mg/L.  Reproduction was reduced in




 DapJ^ia mayna at 250 Mg/L, but not at 140 jzg/L.  A chronic value of 187.1 ^g/L




 was  derived,  and a corresponding acute-chronic ratio of 36.88.  The growth of




 a midge,  Chironomus tentans, was retarded at 230 Mg/L of atrazine,  but not at




 110  ^g/L.   A chronic value of 159.1 Mg/L was calculated, and a  corresponding




 acute-chronic ratio of 4.525 was derived.





       Brook trout,  Salvelinus fontiaalis, had reduced growth at 120 ^g/L, but




 not  at 65  ^g/L,  in a chronic exposure.  A chronic value of 88.32 jUg/L and an




 acute-chronic ratio of -71.33 were calculated.  In two life-cycle tests with




 the  fathead minnow,  Pimephales promelas,  the chronic limits  were set in the




 first test at 213  and 870 ^g/L based upon no adverse effects observed in a




 chronic  exposure at 213 ^g/L and 25 percent mortality in 3-  to  5-day old fry




 at 870 ^g/L.   The  chronic value was 430.5 ^g/L and the acute-chronic ratio was




 34.84.   In the second test,  chronic limits were 250 and 460  ^g/L,  based upon




growth of  larval .fish,  resulting in a chronic value of 339.1 ^g/L  and an




 acute-chronic ratio of 58.98.   The acute-chronic ratio for this species,




 45.33, was calculated as the geometric."mean of^ these two values.   Bluegills,




Lepomis macrochirus,  were unaffected in  a chronic exposure to  95 ^g/L, while




 an equilibrium loss occurred in bluegills exposed to 500 ^g/L.  These were




 established as the  chronic limits, with  a chronic value of 217.9 ^g/L.  Since
                                       54

-------
 the acute value was a "greater than" value, the acute-chronic ratio was

 >36.71.


       Chronic values are available for three Specie3 of saltwater organise.

 The chronic values for Eurytenora affinis ranged from 5,020  to 20,920


 based on survival.  The chronic value for Aaiericamysia ba*ia  was  123.3

 also based on survival.  The chronic value for Cyprinodon variegatus was  2,542

 A/g/L,  based on mortality of juveniles.  The resultant acute-chronic ratio for

 B.  ar-ranis was 2.629, while the acute-chronic ratios for A. Jbahia and C.

 varieg-atus were 8.110 and >6.294, respectively.               ••"-''


       Effect concentrations for freshwater and saltwater plants are lower than

 the acute and chronic values for aquatic animals (Figures ' 4 and 5).  Atrasine

 toxicity to aquatic plants, both algae and macrophytes., commonly  occurs at

 concentrations of 10 ^g/L and above,  with several reports of  toxicity to

 specific plant taxa at concentrations below 10 ^g/L (primarily freshwater

plant  species) .   Effects are thought to be algistatic rather  than algicidal at

 these  lower concentrations, with recovery'occurring once the  atrazine is

removed.  The lowest ECHO values for freshwater green algae with  exposure

durations of 4 days or longer were 10.2 and 4 ^g/L for Cfalamyrfomonas

reinhardtii and Selenastrua cspricomutum,  respectively.  Mean-ECSO values for
               fr                                   "•'.-•
 these  species would be considerably higher.  The lowest reported  EC50 value

for a  freshwater vascular plant species,  Lemna gibba,  was 37  Mg/L in a 14-day

exposure, using wet weight as an endpoint (Figure 4) .


       Conversely, the lowest ECSO based on growth for a saltwater green algae

 species, Weocliloris sp., was 82 ^g/L, while the equivalent value  for a

saltwater vascular plant species, Afyriopiyllum spicatum, was  25 Mg/L.  A  Final

Plant  Value was  not determined, as there were no tests meeting the

requirements stated in the Guidelines (Stephan.et al.  1985).
                                       55

-------
       Atrazine has a limited tendency to accumulate in tissues  of  aquatic




.animals.   BCFs rang.ed from <0.27 to a maximum of 8.5 in  three species of




 freshwater fish.   There are no BCFs available for saltwater  species.




       Aquatic ecosystem structural and functional parameters have  most




 frequently been observed to be adversely affected by atrazine concentrations




 of 10 Mg/L and above (Figures 4 and 5).   However, in a few studies




 (saltwater),  adverse effects have  been observed at lower exposure  levels.  The




 observed effects  have been on both the plant  and animal communities, with the




 effects upon .the  animal community  being secondary in nature, mainly a result




 of decreased  availability of shelter  and, plant matter for food.




       The  effects  data presented above indicate that criteria which adequately




protect aquatic animals and their.uses will probably also, protect  aquatic




plants .and  their, uses.   The national  criteria are determined on the basis of




atrazine toxicity  to  aquatic .animals..  The Criterion .Maximum Concentrations '




for fresh water  (351.2  W/D  and salt .water,  (759.5  ^g/L)  are one-half of the




respective Final Acute  Values  (702.4  and  1,519  Mg/L,  respectively).  These




values are based on Table  1 acute  toxicity values for all invertebrate and




•vertebrate species.   The Criterion  Continuous  Concentrations for fresh water




 (12.35. W/L) and salt water (26.71  W/L)  are  based  on the Final Chronic Values




for fish.   These were calculated by dividing  the Final Fish Acute-Chronic




Ratio  (56.86)  into the  Final Acute  Value  in each case.  .Separate Final Acute-




Chronic  Ratios were determined  for  fish and invertebrates due to the large




differences in the ratios  for the  two  groups.   The  Fish Chronic Values became




the Criterion Continuous Concentrations for fresh water and salt water -because




there  were no -Final -Plant  or Final  Residue Values,  and the results from




studies  in the Other  Data  section did not appear to warrant the establishment




of  a lower concentration at  this time.
                                        56

-------
National Criteria










       The procedures described in  the Guidelines  indicate that,  except




possibly where a locally important apecies is very  sensitive,  freshwater




aquatic  animals and their uses should not be directly affected unaccep.tably if




the  four-day average concentration of atrazine does  not  exceed 12.35 ^g/L more




than once every three years on ,the average, and if  the one-hour average  "




concentration does not exceed  351.2 ^g/L more than  once  every three years on




the  average.   The four-day average of 12.35 pg/L  for the protection of




freshwater animals should also be  protective of most freshwater plants.




       The procedures described in  the Guidelines  indicate that,, except




possibly where a locally important species is very  sensitive,  saltwater




aquatic  organisms and their uses should not be affected  unacceptably if  the




four-day average concentration of  atrazine does not  exceed 26.71 pg/L more




than once every three years on the average, and if  the one-hour average




concentration does not exceed  759.5 Mg/L more than  once  every three years on




the  average.   The four-day average of 26.71 pg/L  for the protection of




saltwater animals should also  be protective of most  saltwater plants.
Implementation
      As discussed in the Water  Quality Standards  Regulation (U.S. EPA 1983a)




and the Foreword to this document,  a water quality criterion for aquatic life




has regulatory impact only when  it  has been  adopted in a State water quality




standard.   Such a standard specifies a criterion  for a pollutant that is




consistent  with a particular designated use.  With the. concurrence of the U.S.




EPA, States designate one or more uses for each body of water, or segment
                                       57

-------
 thereof, and adopt criteria that are  consistent with the use(s)  (U.S. EPA '




 1983a,b, 1987,  1994) .   Water quality  criteria adopted in State water quality




 standards could have the same numerical values as criteria developed under




 Section 304,  of the Clean Water Act.  However,  in many situations States  might




 want to adjust  water quality criteria developed under Section 304 to reflect




 local environmental conditions and human exposure patterns.  Alternatively,




 States may use  different data and assumptions  than the U.S. EPA in deriving




 numeric criteria that  are scientifically defensible and protective of




 designated uses.   State water quality standards  include both numeric and




 narrative criteria.  A State  may adopt a numeric criterion within its water




 .quality standards  and  apply it either state-wide to all waters designed for




 the  use the criterion  is designed to protect or  to a specific site.   A State




may  use an indicator parameter or the national criterion,  supplemented with




other relevant  information,  to interpret its narrative criteria within its




water quality standards when  developing NPDES  effluent limitations under  40



CUF  122.44(d)(1)(vi).2.





       Site-specific criteria may  include not only site-specific criterion




concentrations  (U.S. EPA 1994), but also site-specific,  and possibly




pollutant-specific, durations'of  averaging periods  and frequencies of'allowed




excursions  (U.S. EPA 1991).  The  averaging periods  of  "one hour" and "four




days" were  selected by, the U.S. EPA on the basis  of data concerning how




rapidly some aquatic species react to increases  in the concentrations of  some '




aquatic  pollutants, and "three years- is the- Agency• s  best scientific judgment




of the  average amount of  time  aquatic ecosystems  should be provided between




excursions  (Stephan et>l. 1985; U.S.  EPA 1991).   However,  various species and




ecosystems  react and recover at greatly differing rates.   Therefore,  if




adequate justification  is provided,  site-specific and/or pollutant-specific
                                       58

-------
concentrations, durations, and frequencies may be higher or lower than those




given  in national water quality criteria  for  aquatic life.




       Use of criteria, which have been adopted in State water quality




standards,  for developing water quality-based permit limits and for designing




waste  treatment facilities requires  selection of  an appropriate wasteload




allocation model.  Although dynamic  models are preferred for the application




of these criteria (U.S. EPA 1991), limited data or other considerations might




require  the use of a steady-state model  (U.S.  EPA 1986) .




       Guidance on mixing zones and the design of  monitoring; programs is also




available (U.S. EPA 1987,  1991).
                                       59

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