United States
                     Environmental Protection
                     Agency
 Environmental Research
 Laboratory
 Duluth MN 55804
                     Research and Development
 EPA/600/S3-84/100  May 1985
&EPA          Project  Summary
                     Biological  Availability  of
                     Sediment  Phosphorus  Inputs  to
                     the  Lower  Great  Lakes

                     Scott C. Martin, Joseph V. DePinto, and Thomas C. Young
                       In  this study, river water samples
                     were  collected  from  several  major
                     tributaries to the Lower Great Lakes
                     during  storm runoff events in the
                     spring and early summer of 1980 and
                     1981.  Suspended  sediments   from
                     these samples  were  subjected  to  a
                     chemical fractionation sequence of
                     NaOH-CDB-HCI,  as  well as algal bio-
                     assay analyses of sediment P bioavail-
                     ability using  the Dual Culture  Diffu-
                     sion  Apparatus (DCDA) technique of
                     DePinto.  Sediments from  several of
                     the  bioassay   experiments  were
                     reconcentrated   after the  bioassays
                     and resubjected  to the chemical frac-
                     tionation sequence.  Several  other
                     forms of P inputs to the Lower Great
                     Lakes were also analyzed for chemical
                     composition and/or  bioavailability.
                       This Project Summary  was devel-
                     oped by EPA's  Environmental  Re-
                     search Laboratory, Duluth, MN, to an-
                     nounce key findings of the research
                     project that is fully  documented  in a
                     separate report of the same title  (see
                     Project Report ordering information at
                     back).

                     Introduction
                       Since  April, 1972,  when  the govern-
                     ments of Canada  and  the United States
                     signed the Great  Lakes Water Quality
                     Agreement, extensive  efforts  have  been
                     made to curb eutrophication in the Lower
                     Great Lakes.  This attention  has  been
                     focused  primarily  on   the Lower  Lakes
                     (Erie and Ontario),  where the most severe
                     problems  exist.   Reductions in   the
                     phosphorus (P) loadings to these lakes are
                     generally considered to be the most effec-
 tive means of controlling eutrophication.
 Accordingly,  target total P loads  were
 established based on the reduction  of
 treatment plant effluent P concentrations
 to 1.0 mg/l.  These were later revised
 through  the use of water quality models
 to predict the load reductions necessary
 to achieve the desired  water quality for
 the Great Lakes  (Task  Group  III,  1978).
 Billions of dollars were committed to im-
 proving  P  removal at wastewater treat-
 ment  facilities discharging to the Lower
 Lakes basin.  Although  municipal loads
 have decreased considerably,  the revised
 target loads of 11,000 metric tons/year for
 Lake Erie and 7,000 metric tons/year for
 Lake Ontario  cannot be met via the  pres-
 ent programs. In addition, improvements
 in the water quality of the lakes resulting
 from these load reductions have, to date,
 been less than anticipated. Future courses
 of action being considered include further
 reductions in municipal treatment plant ef-
 fluent P concentrations to 0.5 mg/l and
 improvement  of land management prac-
 tices (e.g. implementation of conservation
 tillage programs)  in order to  reduce dif-
 fuse  (runoff)  sources of P to the  lakes
 (U.S.  Army Corps of Engineers, 1982).
  Consideration of these  options  has
 caused the utility of target loads based on
 total   P  to  be questioned.  It is  now
 recognized that the relative availability of
 P from different  sources for  supporting
 algal  growth must be carefully evaluated
 in  developing  a  cost-effective  manage-
 ment scheme  for the control of eutrophi-
 cation. Nearly 50% of the total P load to
 Lake  Erie and 40% of the Lake Ontario
 load are from diffuse sources.  A large
fraction of this P  is in a  sediment-bound

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form and may,  therefore,  not be readily
available for  an  algal uptake. Studies of
several  Great Lakes  tributaries  have  in-
dicated  that  generally  40%  or  less  of
suspended  sediment total P is potentially
bioavailable.  However,   the   kinetics  of
conversion  of unavailable allochthonous P
to available  forms have  not been  ade-
quately  quantified. It  is  possible that par-
ticulate  material  may settle  out  of  the
water column before  all of its potentially
available P is released.  In addition,  the
rate of  P release from allochthonous par-
ticulate  material  in the water column may
differ considerably from the  regeneration
rate for autochthonous  particulate P.
  As  P associated with suspended par-
ticulate  material enters the  receiving water
environment,  it may become available for
algal uptake  via  desorption,  dissolution,
and  microbial decomposition of organic
matter.   The  degree   of  particulate   P
availability  depends  on  the  physical,
chemical and biological characteristics of
the  particles  and  the   receiving  water.
Among  the most important factors are the
lake's productivity level, soluble  inorganic
P  concentration  in  the  water  column,
temperature,  lake morphometry,  hydrol-
ogy  and mixing dynamics,  size,  shape,
and  density  of  the  particles,  and  the
chemical forms  of P in the  particles.  A
more thorough understanding of the rela-
tionships between these factors  and  the
bioavailability of particulate P, in  order to
predict  P release for a wide variety  of
sediment and receiving  water conditions,
was  the overall goal of  this study.
  The   specific   objectives  of  the  ex-
perimental  program were:
  1.  to  determine the rate and extent of
     sediment P bioavailability for a varie-
     ty  of  tributaries  to the  Lower Great
     Lakes under receiving water condi-
     tions using a bioassay procedure;
  2.  to  physically   and  chemically
     characterize the  tributary suspended
     sediments with respect to factors in-
     fluencing phosphorus bioavailability;
  3.  to  develop a methodology  that may
     be used to estimate the rate and ex-
     tent   of available   P  release from
     suspended  sediments based  on  the
     characteristics of the  sediments; and
  4.  to  compare the  P  bioavailability  and
     chemical characteristics  of  tributary
     suspended  sediments  to other poten-
     tial sources  of  sediment P  to  the
     Lower Great Lakes.


Experimental Approach
  River  water   samples were  collected
from  six Lower  Great  Lakes tributaries
(Maumee,   Sandusky,  Cuyahoga,   and
Genesee Rivers,  and  Honey  and  Cat-
taraugus  Creeks)  during  storm  runoff
events.  Over 50 tributary suspended  sedi-
ment   samples  were  collected  during
March-June, 1980 and  April-June, 1981.
Suspended sediments from these samples
were concentrated and  algal  bioassays of
sediment  P  availability were conducted
using   the   Dual  Culture   Diffusion   Ap-
paratus  (DCDA)  method   of  DePinto
(1982).  In addition, a variety of chemical
and physical analyses were performed, in-
cluding sediment particle size distribution
(results presented in  the full report) and a
chemical fractionation sequence for  sedi-
ment   P.  Kinetic  coefficients  (rate  and
amount) for the release  of  sediment  P
were determined and correlations between
these  values and the physical and chem-
ical parameters were  examined.  This  was
done to develop a means of predicting the
kinetics  of  sediment  P   bioavailability
based  on  a more convenient  analytical
procedure than the sediment  P bioassays.

Results

Tributary Suspended
Sediments
  The P fractionation procedure used  in
this  study  consisted  of   the  sequential
measurement of reactive (R-NaOH-P) and
nonreactive (NR-NaOH-P)  sodium hydrox-
ide  extractable  P,   citrate-dithionite-
bicarbonate extractable  phosphorus (CDB-
P), HCI extractable  P (HCI-P) and the
residual  fraction  after the  above  se-
quence. A  summary  of  the P fractionation
results  for the tributary suspended  sedi-
ment  samples is shown in Table 1.  Con-
centrations  of most of  the extractable
fractions  were consistent  among  the
samples for a  given  tributary.  Greater
variability was seen  between tributaries
than among samples from any one tribu-
tary. Thus,  it appears that temporal  var-
iations  in   the  forms  of  P  bound  to
suspended  sediments in  streams  are  of
minor importance compared to differences
in soil  type and  land  use,  at least for
periods  of  high  flow associated   with
stormwater runoff.
  The  chemical  fractionation  results  in
Table  1 can  provide  insights  into the
forms  and  mobility of  P  associated  with
suspended  sediments from  the different
tributaries.  The sum  of  R-NaOH-P  and
CDB-P fractions may  be  considered  to
represent the concentration of non-apatite
inorganic P (NAIP) in the suspended sedi-
ments,  while HCI-P  provides an estimate
of apatite  P. Thus,  the  Ohio tributaries
contained high levels of  NAIP and small
amounts of apatite  P,  while Cattaraugus
Creek  sediments were  rich  in apatite P
and  low  in  NAIP.  Samples  from  the
Genesee and  Detroit Rivers contained in-
termediate   levels  of  both  NAIP  and
apatite P.
  The release of available P  from  river
sediments  in   the  DCDA  reactors  was
measured  by  monitoring P  uptake by
Se/enastrum  capricornutum in  the  assay
vessel. An example  of  a  plot of cumula-
tive  P release versus time obtained from
bioassays on two samples (#19 and #20) is
shown  in  Figure  1.  The  form of  these
release  curves is typical  of  the  results
observed  for   the  majority  of  samples
analyzed. In Figure 1, the error bars about
each  data  point  represent one standard
deviation for triplicate bioassays, while the
solid  lines  illustrate  the amount  of sedi-
ment P release predicted by the first-order
equation:

      Prel(t) = PultH  - exp(-Krt)]    (1)

where Prei(t) = the amount of sediment P
released at time t (jigP/g  sed),  Puit =  the
total  amount   of sediment  P  ultimately
available for algal uptake (/tgP/g sed), and
kr  =  the first-order  P release rate coeffi-
cient  (day1, base e).
  A   summary  of   the  tributary mean
values calculated for the first-order release
coefficients, Puit and k, are presented in
Table 2. The  order  of  mean bioavailable
sediment P concentrations for  the  trib-
utaries  was  Cuyahoga  >  Maumee >
Honey > Sandusky  >  Genesee  >  Cat-
taraugus. These results confirm that  the
ultimately available  fraction  of tributary
suspended  sediments is  much less than
that  of  autochthonously  produced  par-
ticulate  P (e.g. phytoplankton), while its
release rate is  much  faster.
  The implications  of  not distinguishing
between external ultimately available P
and that produced within the lake in  phy-
toplankton   models  have  been  reported
elsewhere.  When  sediments of  tributary
origin remain suspended in the water col-
umn  of  a  lake for  longer than  about 7
days,  the current  modeling  approach
would tend to overestimate  the  input of
soluble  P  to  the  lake.  For   models
calibrated primarily on  the basis  of  algal
biomass, this  would likely cause  an over-
prediction of  soluble P concentrations in
the water column. In turn, this  might lead
to either an overestimation of the P reten-
tion  time, or an unrealistic adjustment of
other model coefficients in order to effect
a calibration.  It might  also lead  to inac-
curate conclusions about  the relative con-,

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tributions  of  the various P  sources  to
biological productivity in the lake.
  In order to investigate the possibility of
using a  chemical  method  as  a  surrogate
measure of bioavailable P, simple product-
moment correlation  coefficients  (r) were
determined between  all chemically extract-
able sediment  P fractions (Table 1) and
the one-component P  release coefficients,
Puit and  kr (Table 2).
  All of the  chemical  fractions  except
Residual P were significantly correlated (a
< 0.05) with PU|t. The highest correlation
coefficient obtained was between Puit and
non-apatite  inorganic  P  (NAIP   =
R-NaOH-P +  CDB-P);  however, the in-
organic  component  of base-extractable
sediment P (R-NaOH-P)  is more closely
linked  to  bioavailability than  any  other
single chemical fraction  measured  in this
study.  Correlation coefficients and regres-
sion equations  are presented in the  full
report.
  Observation of the changes in sediment
P  fractionation during  DCDA  bioassays
confirmed  that  the major  contribution of
the  various chemically extractable forms
of  sediment  P  to   the  release  during
bioassays came from  R-NaOH-P.  This was
accomplished   by reconcentrating  the
sediments  after a DCDA run and  resub-
jecting  them to the same chemical frac-
tionation  sequence  used  for  initial
characterization.  Seventeen  of  the   52
            tributary samples were  analyzed in this
            manner. The  most  noticeable feature  of
            the  data  from  this  analysis  is  that
            decreases  in  R-NaOH-P  during the
            bioassays  consistently exceeded the
            changes in all other chemical fractions. A
            large and relatively constant percentage of
            the  R-NaOH-P  initially  present in the
            DCDAs was  released  for  all  of the
            samples analyzed. This percentage ranged
            from 66.0  to  80.1%, with  a   mean  of
            70.8%  and a  coefficient of  variation  of
            only 6.5%. In  addition,  AR-NaOH-P was
            very closely  correlated (r  =  0.896)  with
            the  amount  of  sediment P  taken  up  by
            the assay culture in the  bioassays.


            Shoreline Bluff and
            Bottom  Sediment  Samples
              The   erosion   of  shoreline  (bluff)
            sediments  and the  resuspension of lake
            bottom sediments during high winds both
            may result  in  substantial  inputs of par-
            ticulate P into the water column of Lake
            Erie. The biological availability of sediment
            P in samples from these sources was in-
            vestigated  using  chemical  fractionation
            and  bioassay analyses.  The results are
            presented  in  Table  3.  These  data are
            especially interesting  since they encom-
            pass much wider extremes of sediment P
            fractionation  and  bioavailability   than the
            tributary data.
                                            The eroding bluff material contained ex-
                                          tremely low levels of NaOH-  and  CDB-
                                          extractable  P.  In addition,  the T-Sed-P
                                          concentrations of the  Port  Stanlfy and
                                          Rondeau  Park samples  were well below
                                          the  lowest  value measured  for tributary
                                          sediments.  A large percentage  (mean  =
                                          78.3%) of  T-Sed-P  in  the bluff samples
                                          was removed in the HCI  extraction step
                                          and, therefore, was most likely  present in
                                          the  form  of apatite  P.   By comparison,
                                          although  concentrations  (In  ftgP/g sedi-
                                          ment)  were similar for Cattaraugus  Creek
                                          sediments,  HCI P made up only 47.9% of
                                          T-Sed-P.  In DCDA  bioassays  on  t!iese
                                          samples,  none of the sediment  P  became
                                          available to  algae during  the course of the
                                          experiments. This is  consistent with  the
                                          assertion  made  by other researchers that
                                          apatite P  is  virtually unavailable tc aquatic
                                          organisms and that  recessional shoreline
                                          sediments  contain  very  low  levels  of
                                          bioavailable  sediment P.
                                            The Monroe bottom sediment samples
                                          represent the opposite extreme from  the
                                          bluff  sediments.  All concentrations  of
                                          T-Sed-P,  T-NaOH-P, and CDB-P  in  the
                                          Monroe #2  sample were more  than twice
                                          as  large  as  the  highest values observed
                                          for tributary suspended sediments.  Bio-
                                          assays showed that PU|t  for both  Monroe
                                          samples was also over  twice the  largest
                                          value  measured  for  tributary  sediments.
                                          The percentage  of  T-Sed-P available  to
Table 1.    Summary of P Fractionation Results for Tributary Suspended Sediments
Tributary T-SED-P
(TP-TSP)
   SS
 T-NaOH-P
R-NaOH-P
                                                                      Concentrations in ngP/g sediment

                                                                        NR-NaOH-r      CDB-P
                                                          HCI-P
                                                        Residual P
Maumee River (n=9l:
Mean              1,259
C.V.  (%)            14.3

Sandusky River (n =25):
Mean              J, 188
C.V.  (%)            11.7

Cuyahoga River (n = 4):
Mean              1,315
C.V.  1%)            6.2

Cattaraugus Creek (n = 5):
Mean               637
C.V.  (%)            11.0

Honey Creek (n = 41:
Mean              1,189
C.V.  (%)            16.5
Genesee River (n = 1):
                   957
  1,244
  18.9
  1,193
  13.6
  1,246
  9.9
  588
  12.0
  1,218
  14.5
                                  994
Detroit River In = 1)t:
                   1,424
403 (32.01*
   32.4
449 (37.7}
   20.8
575 (43.8)
   13.4
 76(11.9)
   33.2
505(42.5)
   19.1
                                             240 125.1)
              596 (41.9)
273 (21.71
  20.9
298 (25.1)
  20.8
442 (33.6)
  21.4
 46 (7.21
  27.2
356(30.0)
  23.5
                            170(17.8)
              305 (21.41
131 (10.41
  61.9
151 (12.7)
  31.1
133 (10.1)
  29.6
 30(4.7)
  46.5
238 (18.9)
   17.5
237(20.0)
  21.0
280 (21.3)
   16.6
82 (12.9)
  22.7
149 (12.5)      203 (17.1)
   11.3           22.2
                             69 (7.2)
              291 (20.4)
                            187(19.5)
              177(12.4)
 117(9.3)
  32.5
 77 (6.5)
  35.6
1881/4.3!
   6.3
305(47.9)
   12.9
               56 (4.7)
                50.7
                            273 (28.5)
              323 (22.7)
174 (13.8)
  42.2
194(16.31
  42.5
 73 (5.6)
  63.9
 48(7.5)
  8.7
              265 (22.3)
                21.7
                             77(8.0)
              170(11.9!
^Values in parentheses are means expressed as a percentage of T-Sed-P.
^Sediments composited from four sampling locations.

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       400 f
       300 —
       200 —
 CL
 .5
       100
                    A  Sample No. 19 (Maumee R.)
                         , = 0.171 day'\- PuH = 294 ug p/g
                                  Sample No. 29 (Sandusky R.)
Figure 1.
                           Time (days)


Cumulative re/ease of sediment P versus time measured in DCDA biassays on
Samples #75 (Maumee R.) and #20 (Sandusky R.).
algae (over 50%) was greater as well.
Bottom sediments from the Toledo sites,
on the  other  hand, exhibited  chemical
fractionation    and    bioavailability
characteristics  well  within  the  ranges
measured  for  tributary  suspended
sediments.
Sandusky River Bottom
Sediment Samples
  The effect of a point source of P on the
sediment P  chemistry was also briefly ex-
amined.  Our  chemical fractionation  se-
quence  was applied to bottom  sediment
samples from three sites each  upstream
and  downstream from the  Bucyrus, Ohio
wastewater  treatment plant discharge (on
the Sandusky River). The results  are given
in Table 4.  Substantial  increases in  the
R-NaOH-P and CDB-P fractions  occurred
downstream of the  point source. The NR-
NaOH-P, HCI-P, and Residual P fractions
increased by smaller amounts. This move-
ment of P into the sediments is consistent
                               with  the finding that soluble  reactive P
                               concentrations  decreased  rapidly  with
                               distance downstream  from the  Bucyrus
                               treatment  plant  discharge. The  data in
                               Table  4 indicate that, although much of
                               the point  source P may be in relatively
                               unavailable forms such as  CDB-P, HCI-P,
                               and Residual  P, a sizable  amount is also
                               present  as R-NaOH-P,  and is  therefore
                               potentially bioavailable  should  these
                               sediments  become  resuspended and  car-
theless, these  data  suggest that the  P
discharged from an  indirect point source
undergoes a reduction in bioavailability in
transit to the receiving water body relative
to that discharged  from  a  direct point
source.

Conclusions
  An  analytical program  combining  the
accuracy  of bioassay measurements with
the  efficiency  of  chemical fractionation
studies has been shown to be an effective
means of  quantifying sediment P  bioavail-
ability. The information gathered in  this
study  can,  therefore,  serve  as  a  pre-
liminary basis for evaluating the impact of
P  management strategies  on bioavailable
P loads to the  lower Great Lakes. It also
provided  the   necessary   process  ex-
perimental information  for  implementing
changes  in phytoplankton  model  struc-
tures  to  permit   differentiation   of
allochthonous and  autochthonous sources
of bioavailable  P.
  The results presented in  the full report
indicate   that   bioavailability   remains
relatively constant for a given set of soil
type (i.e.  geochemistry and texture)  and
land use  conditions.  Evidence is seen in
the  similarity between the  bioassay  and
fractionation  results  for the Maumee  and
Sandusky Rivers. Also, Cattaraugus Creek
sediments, which  were much  higher in
apatite content, showed a distinctly lower
level  of  P  availability.  The  Cuyahoga
River,  which  experiences  a greater  an-
thropogenic influence, displayed a higher
level  of sediment  P  bioavailability. Little
fluctuation in sediment characteristics was
observed  over  time  for any given tribu-
tary.  Based on these findings,  it is felt
that future monitoring programs should
concentrate on identifying differences be-
tween regions  of distinct  soil type  and
land use.  This  can be accomplished most
readily by using routine chemical extrac-
tion analyses  on  sediments transported
during storm runoff  events supplemented
by  less frequent bioassay  measurements
of sediment P  bioavailability.
                               ried into the receiving water body. Never-
                               Table 2.    Mean First-Order Release Coefficients Calculated from Bioassay Data
Tributary
Maumee River
Sandusky River
Cuyahoga River
Genesee River
Cattaraugus Creek
Honey Creek
T-Sed-P*
in DCDAs
1308
1145
1314
900
559
1198
Ultimately
Bioavailable
Sediment P
337.3
247.1
449.2
173.8
38.8
298.0
Puit as
a % of
T-Sed-P
25.0
21.4
33.9
19.3
7.7
24.9
Release
Rate, k,
(day"1!
0.182
0.177
0.188
0.264
0.131
0.214
                               *The T-Sed-P values listed in this table are based on TP and SS analyses performed on the sediment
                                suspensions placed in the DCDAs.

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Table 3.    Chemical Fractionation and Bioassay Results for Shoreline Erosion (Bluff) and Lake Bottom Sediment Samples

A) Chemical Fractionations (Values in =gP/g sed.):
Sample (Type)
                               T-Sed-P
                                            T-NaOH-P
                                                         R-NaOH-P
                                                                      NR-NaOH-P
                                                                                      CDB-P
                                                       HCI-P
                                                                                                               Residual P
Monroe #1 (bottom)
Monroe #2 (bottom)
Toledo #1 (bottom)
Toledo #2 (bottom)
Port Stanley (bluff)
Rondeau Park (bluff)
L. Superior (bluff)
2266
355J
926
935
279
396
771
1321
1863
208
226
13
8
15
1226
1796
156
187
8
6
13
95
67
53
40
5
2
2
475
612
156
167
94
22
32
258
315
248
227
228
326
543
44
43
105
99
25
28
88
B) DCDA Bioassays (One-Component Release Coefficients Calculated by Thomas Method):



Sample
Monroe t>1
Monroe #2
Toledo tt1
Port Stanley
Rondeau Park

T-Sed-P
in DCDA's
(ngP/gsed.)
2656
3044
947
662
557
Sediment P
Released in
DCDA's
(ngP/g sed.)
964.2
1341.3
99.3
0
0
Ultimately
Bioavailable
Sediment P,
Pu/,(p.gP/gsedJ
1435.0
1482.4
99.3
0
0

Pu/tas a
% of
T-Sed-P
54.0
48.7
10.5
0
0

fie/ease
Rate, kr
(day-')
0.42
0.080
0.140
—
—
Table 4.    Effects of a Point Source Discharge on Phosphorus Fractionation in Sandusky River Bottom Sediments
Miles
from
Site point
no. source
1 9
2 7
3 3
4 0.5
5 5
6 10
T-Sed-P
777
644
470
1510
1039
1404
T-NaOH-P
284
226
55
528
397
596
Concentrations in ngP/g sediment
R-NaOH-P NR-NaOH-P CDB-P
215
177
43
434
320
305
69
49
12
94
77
291
214
160
92
452
308
177
HCI-P
125
123
263
249
165
323
Sandusky River
Residual P Flow
107
91
54
132
119
170
Point
Source
Discharge
References
DePinto, J.V., "An Experimental Appara-
  tus for Evaluating Kinetics of Available
  Phosphorus  Release from Aquatic Par-
  ticulates." Water Res.,  16,  1065-1070
  (1982).
Task  Group III, "Fifth  Year Review  of
  Canada-United  States  Water  Quality
  Agreement." Report of Task Group  III,
  A Technical  Group to  Review Phospho-
  rus  Loadings,   U.S.   Department  of
  State, Washington,  D.C. (1978).
U.S. Army Corps of Engineers, "Lake Erie
  Wastewater  Management Study - Final
  Report."  Buffalo, N.Y. (1982).
S. C. Martin, J.  V.  DePinto,  and  T. C. Young are with Clarkson College of
  Technology, Potsdam, NY 13676.
W. L. Richardson is the EPA Project Officer (see below).
The  complete report, entitled "Biological Availability of Sediment Phosphorus
  Inputs to the Lower Great Lakes," (Order No. PB 85-121 036; Cost: $17.SO,
  subject to change) will be available only from:
        National Technical Information Service
        5285 Port Royal Road
        Springfield,  VA 22161
        Telephone: 703-487-4650
The EPA Project Officer can be contacted at:
        Large Lakes Research Station
        Environmental Research Laboratory—Duluth
        U.S. Environmental Protection Agency
        Grosselle.MI48138
                                                                                   U. S. GOVERNMENT PRINTING OfFICE: 1985/559 111/10846

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