United States
 Environmental Protection
 Agency
 Robert S. Kerr Environmental
 Research Laboratory
 Ada OK 74820
Research and Development
 EPA-600/S2-83-073  Jan. 1984
Project Summary
 Determination  of  Activated
 Sludge  Biokinetic  Constants
 for Chemical  and   Plastic
 Industrial Wastewaters
Don F. Kincannon and Enos L Stover
  The most widely used method of
wastewater treatment is biological
treatment. The use of kinetic models to
describe the behavior of  a biological
wastewater treatment process has
become widely accepted practice. The
most often used kinetic models include
those developed by Eckenfelder, McKin-
ney, Lawrence and McCarty, and Gaudy.
Eckenfelder (first model) and McKinney
use first order  kinetics for substrate
removal; Lawrence and McCarty, and
Gaudy use the empirical Monod kinetics
for substrate removal; and Eckenfelder
(second order)  relates the substrate
removal rate as a function of the
remaining substrate concentration to
the initial concentration. All  of  these
models contain kinetic constants and
the usefulness of each  model is a
function of the reliability of the kinetic
constants. However, there has.not been
enough information available to establish
reliable values for these kinetic constants
for industrial wastewaters.
  Cooperative Agreement CR 806843-
01-02 has determined the biokinetic
constants and fate for 24 toxic organic
pollutants when present  in a highly
biodegradable wastewater. Biokinetic
constants were also determined for
eight groups of three  chemicals. The
biokinetic constants were  determined
based upon biochemical oxygen demand
(BODs), total organic carbon (TOC),
chemical oxygen demand (COD), and
specific organic chemicals. The normal
approach for determining biokinetic
constants has been to plot the average
values. This approach masks the actual
 scatter of the data  and provides an
 average value or 50  percent probable
 value for the biokinetic constants to be
 used in design. This study has produced
 a methodology for analysis of the test
 results from biological activated sludge
 systems for determining the biological
 variability inherent in these types of
 systems.
  This study also investigated the
 possibility of predicting the fate and
 effluent concentrations of the various
 priority pollutants. It was  found that
 good predictions for the priority pollu-
 tants can  be made. In general, it can be
 predicted that nitrogen compounds,
 phenols, oxygenated compounds, poly-
 nuclear aromatics, and  phthalates will
 be removed by biological degradation.
 Aromatics will  be removed  by both
 biological degradation • and stripping,
 except when the Henry's Law Constant
 is in the 10~5 atnvmVmole range, then
 that chemical will  be removed by
 biological degradation only. Halogenated
 hydrocarbons present a special problem.
 Some are removed by stripping, whereas,
 others are removed by combined biode-
 gradation  and stripping. No parameter,
 such as  Henry's Law Constant or
 partition coefficient,  was found that
 would predict which removal mechan-
 isms would persist. Therefore, biode-
 gradability studies must be conducted.

  This Project Summary was developed
by EPA's Robert S. Kerr Environmental.
Research Laboratory. Ada.  OK. to
announce  key findings of the research
project that is fully documented in a

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separate report of the same title (see
Project Report ordering information at
back).
Introduction
  Provisions in  the Clean Water Act
clearly require the  U.S. Environmental
Protection Agency to promulgate regulations
to protect streams  and  publicly owned
treatment works (POTW's) and to see to it
that regulatory practices are in place to
protect the vast  investment of  public
funds. This law  can affect nearly all
industrial  manufacturing plants now
discharging or planning to discharge into
POTW's or a stream.  An  important
category for which effluent guidelines are
needed  is  waste  components  from
chemical and plastic industries. For this
category,  the problem is  extremely
difficult because of the thousands of
different chemicals  produced at  plants
across the  nation. The product lines of
these plants, and thus the nature of the
wastes, are not fixed and change with
market needs.  This industrial activity
consequently produces wastewaters of
great and changing variety in the types of
pollutants they  contain.  Furthermore, a
significant number of the growing list of
priority pollutants is  appearing in various
waste  streams from the manufacturing
of chemicals and plastics.
  Because of the immensity and tremen-
dous variety of  products and process
routes from production, it is logical that a
computer programming procedure would
be developed to handle the definition of
wastewater composition as well as
alternate  processing  routes for  the
treatment  of wastewater. Computer
programs have been developed that  can
combine waste load information on each
product, select various product treatability
routes, combine them, design and cost-
out  a  plant's treatment system, and,
finally, allocate the  estimated treatment
costs to the contributing  product process
route. However, before plant costs can be
modeled, the treatment design models
must be established. Design models are
available but the biokinetic parameters
for the chemical and plastic industries are
not available. Therefore, the determination
of the  biokinetic parameters becomes of
vital concern to the success of the cost
model.
  The  current best  practical technology
(BPT) for treatment of chemical industrial
and plastic  manufacturing wastes usually
involves activated sludge, since biological
treatment is often found to be the most
cost-effective in dealing with relatively
low concentrations of the types of organic
materials in such wastes.
  Modern approaches for the design of
activated sludge employ mathematical
process models depicting the relationships
among factors affecting the kinetics of
wastewater purification. All design models
require quantitative assessment of
numerical values for biokinetic constants
for  the activated sludge in  question.
These values are, in general, a function of
the type of carbon source comprising the
carbonaceous biochemical oxygen demand.
Various models are available for use in
designing activated sludge processes.
  The primary objective of this study was
to determine the biokinetic constants
needed in process modeling for predicting
effluent quality and design configurations
of activated sludge processes treating the
waste streams from chemical and plastic
manufacturing. Secondary objective was
to refine the method  of combining  the
biokinetic constants of components in a
mix  in order to determine the  overall
biokinetic  constants of the mix  and to
establish the fate of the priority pollutants
in the wastewater.

Conclusions
  High treatment efficiencies (in terms of
biochemical oxygen demand, BODs; chemi-
cal oxygen demand, COD; and total organic
carbon, TOC) can be achieved by biological
treatment  of  wastewaters containing
toxic priority pollutants.  This study  has
shown that priority  pollutants  can be
accommodated in biological wastewater
treatment processes. In addition, it was
found that many of the priority pollutants
were biodegraded to very low concentra-
tions. In other cases, the priority pollutants
were stripped to low  levels without
interfering with the biodegradation of the
other organics.
  The biokinetic constants of the waste-
waters containing the priority pollutants
are representative of a readily biodegrad-
able .wastewater. The biokinetic constants
are subject to variability, and a frequency
analysis should be used in determining the
biokinetic constants. A new design model
provides  an  alternative method  for
accounting for  the variability  of  the
system. This new model provides designs
comparable with the other design models
without the variability of the biokinetic
constants.
  The fate and effluent concentrations of
priority pollutants can be predicted with a
high degree  of  accuracy. Parameters,
such as Henry's Law Constant and Log
partition coefficient, are key elements in
predicting the fate of  priority pollutants.
Some priority pollutants are both biode-
graded and stripped. These  are  more
difficult  to predict. Some means  of
estimating the biodegradability of priority
pollutants is needed.

Recommendations
  This study has investigated the biological
treatment of 24 toxic organic chemicals.
The study also included eight groups of
three chemicals  as a mixture. It  is
recommended that additional research
be conducted using chemicals that would
enhance sorption as a removal mechanism.
Biodegradability  and fate  of  additional
chemicals could allow the creation of a
structure/activity correlation for predict-
ing the degradability and fate  of toxic
organics. Cost impacts for the correction
of existing  problems and  the ability to
anticipate adverse environmental impacts
from  the manufacture of chemicals
would be made available through the use
of such correlation.
  It would also be of benefit to investigate
the effects of  factors other than the
priority  pollutants. Surface tension,
sulfur compounds,  inorganics, etc. may
be much more  detrimental to biological
treatment than, so called, toxic organic
chemicals.
  It  is recommended that the water
pollution control  field recognize that
"toxic" organic chemicals can be treated
in a biological wastewater  treatment
plant with  no detrimental  effects to the
treatment process.
Experimental Methods
  The general experimental plan specified
bench-scale continuous  flow activated
sludge reactors be  used  to  treat  a
synthetic wastewater containing  selected
chemicals normally present in chemical
industry  wastewaters. The bench-scale
activated sludge system is  shown in
Figure 1. The system  consisted  of  a
stainless steel  internal  recycle  reactor.
The activated sludge  reactor had  a
volume of 3.0 liters and the  settling
compartment had a volume of 3.23 liters.
The reactor and settling compartment
both had a stainless steel  cover. The
wastewater was pumped from a sealed
feed tank to the reactor. The effluent from
the settling unit flowed  by gravity to a
collection tank. The off-gas was pulled by a
vacuum  pump through a  purge trap
which  contained 6 inches of Tenax and 4 •
inches of Silica Gel. The  influent air and
off-gas were  measured with  air-flow
meters. The influent wastewater flow
was regulated to provide a hydraulic

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                                 Off-Gas
                                           D^^~~^F,ow   VK""m
                                           Purge Trap    Meter     ™mp
                                             Effluent
                                           Collection Tank
 Wastewater
 Feed Tank
Figure 1.
    Pump

Bench-scale activated sludge pilot plants.
detention time of 8 hours in the activated
sludge reactor. All parts of the experimental
apparatus were constructed of materials
to minimize contamination.
  Two types of experimental studies
were conducted with the previously
described stainless  steel  reactors.  A
nonbiological system  was  operated  to
determine the strippability of the chemical
compound. In this system, the reactor and
settling  compartment were filled with
distilled water. The feed tank contained
the wastewater.  The wastewater was
pumped to the reactor and the level  of
TOC and  specific compound present  in
the reactor was observed as a function of
time.  When the wastewater had been
completely diluted-in,  influent, effluent,
and off-gas  samples  were collected for
specific compound analyses.
  The second experimental system was a
biological activated sludge system.
Activated  sludge for initial seeding was
obtained from a local municipal activated
sludge plant. Three individual  systems
were acclimated to  each synthetic
wastewater containing the pollutant to be
evaluated. The activated sludge systems
were  operated  at mean  cell residence
times (SRT) of  2, 4,  and 6 days. After
acclimation,  influent,  effluent,  mixed
liquor and off-gas samples were collected
over a 60-day period for  analyses. All
three  reactors  were operated at  a
hydraulic detention time of 8 hours.
                             The synthetic wastewater referred to
                           as the "base  mix" contained  ethylene
                           glycol,  ethyl alcohol, glucose,  glutamic
                           acid, acetic acid,  phenol,  ammonium
                           sulfate, phosphoric acid, and salts. The
                           specific chemical compoundto be studied
                           was added to this base mix. The specific
                           compound and the corresponding initial
                           BODs,  TOC,  and  COD  are shown  in
                           Table 1.

                           Results and Evaluation
                             Treatment efficiencies when measured
                           in terms  of BOD5,  TOC,  and COD were
                           very high for all systems in this study. The
                           mean BODs did not exceed 10.0 mg/l for
                           any system. Generally, the  mean BOD5
                           was below 5.0 mg/l. This indicates that
                           the "base mix" was highly biodegradable.
                           This factor did  create problems in
                           obtaining data  for determining biokinetic
                           constants. The ideal  situation  is one in
                           which the effluent BODs varies over a
                           wide range. However, with the wastewa-
                           ters used in this study, even an SRT of 2
                           days produced a very low effluent BODs.
                           Therefore, SRT values of 2,4, and 6 days
                           produced effluent BODs's with very little
                           difference. The high treatment efficiencies
                           also indicated that the priority pollutants
                           used in this study had very little or no
                           effect on the biological treatment of the
                           wastewater.  Even when the priority
                           pollutant  was not biodegraded,  it did not
                           affect the treatment efficiency.
  It was found that the specific chemicals
were  removed by several  mechanisms.
These included stripping, biodegradation,
and sorption. The percent removed by
each mechanism is shown in Table 2.
  The normal approach for analyzing and
presenting biological system data is to
plot the average values at each //„ or SRT.
This approach masks the actual scatter of
the data and provides an average value or
50 percent probable  value  for the
biokinetic constants to be used in design.
Use of these biokinetic  constants will
provide a system capable of achievingthe
desired effluent quality only 50 percent of
the time. The allowed statistical variability
of the effluent discharge criteria  must be
evaluated to determine  the appropriate
biokinetic constants to use for design.
Designing to meet a specific effluent
quality 95 percent of the time will require
a significantly larger reactor volume than
that required for 50 percent of  the time
and will require use of  the 95  percent
probable biokinetic constants, as shown
in the following example.
  Using Eckenfelder's design model and
the following design  conditions the
impacts of data variability for the acryloni-
trile - containing wastewater  can be
demonstrated.

  F = 10MGD
  S, = 200 mg/l BOD5
  Se = 5mg/l BODs
  X =  1000 mg/l  volatile suspended
       solids

The activated sludge reactor volumes are
determined as follows:
= F
                    , - Se)
                X Ke' Se

Using the 50 percent line or Ke' = 31, the
required  reactor volume will be:

      v = 10x106x200x(200-5)
              1000x31 x5

      V= 2,516,000 gallons   .


This reactor volume of 2,516,000 gallons
should provide an effluent soluble BOD5
(Se) of 5 mg/l 50 percent of the time.
Using the 95 percent line or Ke' = 11, the
required  reactor volume will be:

      v= 10 x 106x200x(200-5)
             1000 x 11x5

      V=  7,091,000 gallons

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Table 1.    Wastewater Concentrations
Group
Chemical
    Wastewater concentrations, mg/l
    BOD5           TOC        COD
1



II



III



IV



V



VI



VII



VIII



Tetrachloroethane
Nitrobenzene
2,4 dichlorophenol
Combined
Acrylonitrile
Acrolein
1,2 dichloropropane
Combined
Benzene
Methylene chloride
Ethyl acetate
Combined
1,2 dichlorobenzene
1,2 dichloroethane
Phenol
Combined
2,4 dinitrophenol
1,3 dichlorobenzene
1,1,1 trichloroethane
Combined
Pentachlorophenol
Trichloroethylene
Bis (2-ethylhexyl) phthalate
Combined
Toluene
Ethylbenzene
Phenanthrene
Combined
Chloroform
Carbon tetrachloride
Naphthalene
Combined
244
255
298
314
192
219
247
279
212
287
259
248
162
201
277
268
245
178
244
227
265
295
271
270
180
236
270
217
240
257
219
317
224
253
234
246
189
190
199
197
132
170
145
133
152
187
163
176
203
136
183
143
153
179
159
169
129
143
166
147
155
162
163
170
496
668
572
574
483
477
480
565
441
480
450
400
416
460
486
454
551
379
530
429
470
530
528
574
416
520
510
470
363
387
390
409
Control
            "Base Mix"
                                             266
                                                            170
                                                                       460
This reactor volume of 7,091,000 gallons
is almost three times the previous reactor
volume and should provide an effluent
soluble BOD5 (Se) of 5 mg/l 95 percent of
the time. Using the 75 percent line or Ke' -
19, the required reactor volume will be
4,105,000 gallons. If the reactor volume
was designed at 2,516,000 gallons, the
following probability of occurrence of Se
could be expected:
  Probability Level %
         50
         75
         95
     Se (mg/l)
         5
         8
        14
  The variability analysis  approach
presented indicated  that Kd was a true
biokinetic constant, //max was also treated
as a true biokinetic constant, and since
/AT»X = K-Yt, and Yt  was found to  be a
variable biokinetic coefficient, jumax and K
could not both be treated as constants. If
Yt = yUmax/K varies, yumax, K, or both must
vary.  If both  /jmax and K  vary,  the
prediction of these coefficients becomes
more difficult. The decision to make//max a
biokinetic  constant and K a variable
biokinetic coefficient depending on the
variable biokinetic coefficient Yt simplifies
the prediction of (ime* and K. The following
reasoning was used in the selection of
/Umax as a biokinetic constant.
  K is simply the maximum U, and U was
found to vary at constant//,, or/j. At/Umax, U
would be equal to  K and could  be
expected to  be variable. Also  in a
heterogeneous population such as in the
activated sludge process treating a
particular wastewater, several different
kinds of  bacteria  will actually be in
predominance;  however,  shifts in the
ratios of these specific micro-organisms
in predominance can change. As these
population  dynamic  shifts occur Yt
changes, and thus,  the  mixed liquor
solids (X) changes. The specific substrate
utilization rate (U) could be expected to
change as  X changes; however, /umax
should not change. As //„ or /u approaches
A/max, the microorganisms with lower /umax
values than the existing fjn or ^ will be
washed out  of the system. Therefore, the
microoranisms with the greatest /Umax will
predominate at the /umax condition of the
system. At this absolute //max which could
be considered to be a constant,  many
microorganisms will  have been washed
out of the system, but several microorgan-
isms will still remain. These few organisms
may still provide a variable maximum U or
K value but with lower variability since
the few  remaining  organisms should
provide less variability.  All  the  other
substrate utilization terms (Ks and Ke' or
Ke) were also found  to be variable in a
manner similar to the true yield. Even at
very constant controlled /un's or SRT's, all
the substrate  utilization characteristic
descriptors were  found to be variable.
Based on this reasoning, Yt Ke', Ke, Ks and
K  are  all  considered to be variable
biokinetic coefficients, while Kd and fjmax
are considered to be biokinetic constants.
The variability observed in these biokinetic
coefficients during the treatability study
can and should  be considered during
scale-up design  to achieve specific
effluent discharge criteria.
  In addition to the variability analysis for
the existing models, a new model has
been presented. This model provides an
approach in which there is no variability
of the biokinetic constants. All variability
can be handled as influent variability and
normal procedures can be used. The new
design model provides  design values
comparable with the other design models.
  This study  has also  looked  at the
possibility  of  predicting  the  fate and
effluent  concentrations  of the various
priority pollutants. It has been found that
good predictions for the priority pollutants
can  be  made. In  general,  it can  be
predicted  that  nitrogen compounds,
phenols,  oxygenated compounds,  poly-
nuclear aromatics, and phthalates will be
removed by biological degradation.
Aromatics will be  removed by  both
biological  degradation and  stripping,
except when the Henry's Law Constant is
in the 10~5 atm-rnVmole range, then that
chemical will  be  removed by biological
degradation only. Halogenated hydrocar-
bons present a special problem. Some are
removed by stripping,  whereas, others
are removed by combined biodegradation
and stripping.  No  parameter,  such as
Henry's Law Constant or  partition coeffi-
cient, was found that would predict which
removal  mechanisms would  persist. It
was also found that the type of reactor
could be a factor. Therefore, biodegradabil-
ity studies must  be conducted.  If  the
biodegradability is high, the chemical will
be both stripped and biodegraded.
   It was  also found that biodegradability
and stripping constants could be developed
for each priority pollutant and used very
successfully  in  predicting the effluent

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Table 2.    Removal Mechanisms of Toxic Organics
                                         Percent Treatment Achieved
                                   Single Units
                               Combined Units
Compound
Strip. *    Sorption   Biol,H
Strip.  Sorption     Biol.
Nitrogen Compounds
 Acrylonitrile                                       99.9
Phenols
 Phenol                                           99.9
 2,4-DNP                                          99.3
 2,4-DCP                                          95.2
 POP                                      0.58     97.3
Aromatics
 1,2-DCB                       21.7               78.2
 1.3-DCB
 Nitrobenzene                                      97.8
 Benzene                        2.0               97.9
 Toluene                        5.1       0.02     94.9
 Ethylbenzene                    5.2       0.19     94.6
Halogenated Hydrocarbons
 Methylene Chloride              8.0               91.7
 1,2-DCE                       99.5       0.50
 1.1.1-TCE                     100.0
 1,1,2.2-TCE                    93.5
 1,2-DCP                       99.9
 TCE                           65.1       0.83     33.8
 Chloroform                    19.0       1.19     78.7
 Carbon Tetrachloride            33.0       1.38     64.9
Oxygenated Compounds
 Acrolein                                          99.9
Polynuclear Aromatics
 Phenanthrene                                     98.2
 Naphthalene                                      98.6
Phthalates
 Bis(2-Ethylhexyl)                                   76.9
Other
 Ethyl Acetate                    1.0               98.8
                               2.0
                               6.2
                               7.5

                               2.0
                              75.0
                              99.7
                              96.2
                              99.5
                              69.7
                              15.0
                              28.0
        0.30
        1.36
        0.03
                  90.0

                  99.9
                  99.2
                  77.1
                  97.8

                  99.9

                  33.8
                  97.2
                  93.7
                  92.4

                  97.2
                  25.0
30.1
82.8
77.7

90.0

94.7
78.6

85.7

98.9
 *Strip. = Stripping.
 **Biol. = Biological.

concentrations for the  pollutants when
combined with other priority pollutants or
when the influent concentrations varied.
                Don F. Kincannon  and Enos L Stover are  with Oklahoma State  University,
                  Stillwater. OK 74078.
                Thomas E. Short is the EPA Project Officer (see below).
                The complete report consists of two parts, entitled "Determination of Activated
                  Sludge Biokinetic Constants for Chemical and Plastic Industrial Wastewaters,"
                  (Order No. PB 83-245 233; Cost: $14.5O) and
                  "Determination of Activated Sludge Biokinetic Constants for Chemical and
                  Plastic Industrial Wastewaters (Appendix A—Raw Data)," (Order No. PB 83-
                  245 241; Cost: $25.00, costs subject to change)
                The above reports 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:
                        Robert S.Kerr Environmental Research Laboratory
                        U.S. Environmental Protection Agency
                        P.O. Box 1198
                        Ada, OK 74820

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United States
Environmental Protection
Agency
Official Business
Penalty for Private Use $300
Center for Environmental Research
Information
Cincinnati OH 45268
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                                                                                       {, U.S. GOVERNMENT PRINTING OFFICE: 1984-759-102/825

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