EPA-R2-73-279
July 1973              Environmental Protection Technology Series
Phosphorus Removal
By  Trickling Filter Slimes
                                 Office Of Research And Development

                                 U.S. Environmental Protection Agency
                                 Washington, D.C. 20460

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            RESEARCH REPORTING SERIES
Research reports of the  office  of  Research  and
Monitoring,  Environmental Protection Agency, have
been grouped into five series.  These  five  broad
categories  were established to facilitate further
development  and  application   of   environmental
technology.   Elimination  of traditional grouping
was  consciously  planned  to  foster   technology
transfer   and  a  maximum  interface  in  related
fields.  The five series are:

   1.  Environmental Health Effects Research
   2.  Environmental Protection Technology
   3.  Ecological Research
   4.  Environmental Monitoring
   5.  Socioeconomic Environmental Studies

This report has been assigned to the ENVIRONMENTAL
PROTECTION   TECHNOLOGY   series.    This   series
describes   research   performed  to  develop  and
demonstrate   instrumentation,    equipment    and
methodology  to  repair  or  prevent environmental
degradation from point and  non-point  sources  of
pollution.  This work provides the new or improved
technology  required for the control and treatment
of pollution sources to meet environmental quality
standards.

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                                              EPA-R2-73-279
                                              July 1973
PHOSPHORUS REMOVAL BY' TRICKLING FILTER SLIMES
                     by

                A.  E.  Zanont
              Grant #17010 DZG
               Project Officer

               Edwfn F. Barth
    U.  S.  Environmental Protection Agency
   National  Environmental  Research Center
           Cincinnati, Ohio 45268
                   for
      OFFICE OF RESEARCH AND MONITORING
    U. S.  ENVIRONMENTAL PROTECTION AGENCY
            WASHINGTON, D.  C. 20460

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                EPA Review Notice
This report has been reviewed by the Environmental
Protection Agency and approved for publication.
Approval does not signify that the contents neces-
sarily reflect the views and policies of the
Environmental Protection Agency, nor does mention
of trade names or commercial products constitute
endorsement or recommendation for use.
                       11

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                             ndSTRACT
     At the time this  investigation was started almost ail the reported
work in the I  fterature on the biologi'cal removal of phosphorus had been
with the activated sludge wastewater treatment process.  Little informa-
tion was available on the trickling filter performance in this regard.
This is surprising since there are many uni'ts of this type i'n operation
in the country today.  The published data plus the writer's own experi-
ence Indicated phosphorus removal for the trickling filter process from
negligible amounts to as much as 20 per cent under currently operating
schemes.

     A review of recent investigations on the behavior of fixed biologi-
cal slimes plus a degree of similarity between slfme biota suggested
that the potential for phosphorus removal by trickling filters, and
other processes using slimes growing on solid surfaces, Is higher than
the values which have been reported.  The primary objective of this
research project was to explore the possibility of greater phosphorus
uptakes by biological slimes growing on a solid surface as the result of
varying such conditions as slime thickness, feed constituents, growing
environment, etc.

     Two laboratory units were built In an attempt to study phosphorus
uptake by biological slimes.  The first consisted of four parallel discs
rotating within separate reaction vessels.  Slime was grown on the discs
under many different operating conditions with little success in obtain-
ing "luxury" uptake of phosphorus.  The disc slimes usually contained
1.5 to 2.5 per cent phosphorus on a dry weight basis.  In the cases
where the values were above this range, ft was usually because of the
presence of the calcium added to the feed.

     The other unit consisted of an inclined surface which contained
four one-half  inch wide channels of varying lengths for growing slimes.
Provisions were included for varying the slope and subjecting the slime
surface to ultraviolet irradiation.  Basically the results of the test
conducted on this apparatus demonstrated that phosphorus uptake on a
slfme surface  Is almost entirely due to a biological  mechanism with
physical  adsorption playing a very minor role.

     This report was submitted in fulfillment of Project Number 17010 DZG
under the sponsorship of the Environmental  Protection Agency by the
Department of  Civil  Engineering, Marquette University,  Milwaukee,  Wisconsin
53233.   The Project Engineer was A. E.  Zanoni.
                                Ul

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                         CONTENTS

Section                                                              Page
   1     Conslusions                                                   1
  11     Recommendations                                               3
 111     Introduction                                                  4
  IV     Literature Review                                             7
   V     Base Objectives os Study                                      9
  VI     Analytical Procedures Employed                               11
 VII     Analysis of Actual Trickling Filter Slimes                   18
VIII     Design and Construction of Laboratory Disc Apparatus         22
  IX     Tests Conducted with Laboratory Disc Apparatus               25
   X     Design and Construction of Laboratory Channel Apparatus      31
  XI     Tests Conducted with Laboratory Channel Apparatus            34
 XII     Discussion of Results                                        39
XIII     Acknowledgements                                             50
 XIV     References                                                   51
  XV     Appendix                                                     54

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                           FIGURES
Figure No.                   Title

     I              Schematic of Laboratory  Disc                    23
                           Apparatus

     2             Photographs  of Disc Apparatus                    24

     3             Schematic of Channel  Apparatus                   32

     4             Photograph of Channel  Apparatus                  33
                             vf

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                                TABLES
Table
  4


  5
  8
  10



  II

  12


  13
             Title

Analysis of Trickling Filter Slimes
    From Six Plants In the Milwaukee
    Area

Effect of Refrigerated Storage of Waukesha
    Trickling Filter Slime on Phosphorus
    Level

Effect of Room Temperature Storage of
    Actual Trickling Filter Slimes on
    Phosphorus Level

Effect of 35°C Storage of Actual Trickling
    Filter Slimes on Phosphorus Level

Effect of Anaerobic Storage at 35°C of
    Waukesha Plant Slime on Phosphorus
    Levels (Run No. I)

Effect of Anaerobic Storage at 35°C of
    Waukesha Plant Slime on Phosphorus
    Levels (Run No. 2)

Effect of Anaerobic Storage at 35°C of
    Laboratory Slime on Phosphorus
    LeveIs

Effect of Anaerobic Storage at 35°C of
    Waukesha Plant Slime on Phosphorus
    LeveIs

Effect of Anaerobic Storage at 35°C of
    Waukesha Plant Slime on Phosphorus
    LeveIs

Effect of Anaerobic Storage at 35°C of
    Laboratory Slime on Phosphorus
    LeveIs

Total Hardness of Trickling Filter Slimes

Log of All Test Runs Conducted with Disc
    Apparatus

Analysis of Disc Slimes at Varying Speeds
    and Different Growing Conditions
Appendix - 55



Appendix - 60



Appendix - 61



Appendix - 64


Appendix - 66



Appendix - 68



Appendix - 70



Appendix - 71



Appendix  - 73



Appendix  - 76



Appendix  - 78

Appendix  - 79


Appendix  - 89
                                    vl

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                              TABLES (Continued)
Table
   14
             Title
 Page
   16


   17


   18


   19


   20

   21

   22

   23

   24
Percent Calcium and Magnesium In Disc
    S11mes

Analysis of Channel Slimes Preliminary
    Runs

Feed and Effluent Analyses RemovaI-Storage
    Comparison Runs on Channel Apparatus

Analysis of Channel Slimes RemovaI-Storage
    Comparison Runs

Feed and Effluent Analyses Ultraviolet
    Studies on Channel Apparatus

Feed and Effluent Analyses Starve-K!11
    Study on Channel Apparatus

Multiple Linear Regression Analysis-Disc I

Multiple Linear Regression Analysis-Disc II

Multiple Linear Regression Analysis-Disc III

Multiple Linear Regression Analysis-Disc IV

Summary of Multiple Correlation and F Values
    on Multiple Linear Regression Analysis
    of Data From Disc Apparatus
Appendix


Appendix


Appendix


Appendix


Appendix


Appendix


   41

   42

   43

   44

   45
104


109


110


114


117


119
                                    vill

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                         SECTION I

                        CONCLUSIONS
 !.  For the analysis of total phosphorus, the ashing method proved to
    be superior to the persulfate method for sludges and semi-sol ids
    like biological slimes.  Due to its simplicity and accuracy the per-
    sulfate method is still recommended for the routine analysis of
    water and wastewater samples.

2.  A satisfactory method was developed for estimating the per cent
    carbon in slime samples by using a modified COD procedure.

3.  A large number of slime samples from six different trickling filter
    plants in the Milwaukee area were analyzed with the results that
    the volatile solids were mostly in the 70 to 80$ range, phosphorus
    in the 2 to 3$ range, nitrogen in the 6 to Q% range and carbon in
    the 40 to 50/S range.

4.  A number of tests were conducted In which slimes from actual trick-
    ling filters were incubated under anaerobic conditions and the
    phosphorus release properties were examined.  Initially a small
    amount of phosphorus Is released to the surrounding liquid but the
    bulk of it remains tied up in the solid fraction.  This indicates
    that most likely most of the phosphorus in actual slimes is tied up
    in chemical  precipitates.

5.  Slfmes were grown on the disc apparatus under varying feed and en-
    vironmental  conditions.  The amount of total phosphorus in the
    slime varied from approximately 0.3 to 3.0$ for all the test runs
    conducted.  The low values occurred under conditions of phosphorus
    deficiency in the feed, whereas the high value usually occurred
    when some calcium hardness was added to the feed.

6.  The results of the disc study indicated that there Is little like-
    lihood of Increasing the amount of biologically stored phosphorus
    much over 2.0 to 2.5 percent.   This means for most municipal waste-
    waters, a phosphorus removal  of less than 20$ will  usually be
    achieved under ideal conditions, considering average hardness and
    the present phosphorus levels in raw wastewater.  Chemical addition
    will be required to increase phosphorus removals above this level.

7.  It Is difficult to control the biological removal of phosphorus by
    slimes at any given level, which means the biological  phenomenon In-
    volving phosphorus uptake Is much more random or unpredictable than
    the better known carbon removal  kinetics.

8.  Through the use of the channel apparatus a clearer Insight as to
    the mechanism of phosphorus uptake on a slime surface was obtained.

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The mechanism was shown to be almost entirely due to biological  ac-
tivity.  Physical adsorption which is very important In the soil
regime, is of little Importance when it comes to slimes growing  on
solid surfaces.   Had physical adsorption proven to be more signifi-
cant, this would have had interesting implications in the design of
treatment units  using biological slimes.

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                            SECTION  I I

                          RECOMMENDATIONS
     At the time that this study was proposed a number of papers had
occurred  in the literature purporting that luxury biologfcal storage
of phosphorus was possible in the activated sludge process under
certain plant operational and environmental conditions.  Those observa-
tions plus the fact that little was known about the trickling filter
process in this regard served as the primary impetus for this study.

     It has since been fairly well established that the luxury uptake
of phosphorus in the activated sludge process was probably due to
chemtcal precipitation with the normal hardness-causing cations under
favorable pH conditions.  It is the conclusion of this study that the
biological removal  of phosphorus by slimes growing on sol I'd surfaces
is also quite limited, simply because the quantity of biologically
fixed phosphorus in slime on a dry weight basis will not exceed 2.5%
regardless of environmental  and operating conditions.  Phosphorus must
be incorporated into the sludge if it is to be removed from a waste-
water.   The only practical  way of achieving this in the trickling
filter process is by chemical addition.   The results of this study
strongly suggest that further investigation along the lines of this
reported study not be encouraged.  The only situation in which biologi-
cal removal  of phosphorus might be worth considering is the case of a
wastewater with a high carbon to phosphorus ratio.   Even under this
ideal case it will  probably be found that the control of the operation
wiI I  be quite difficult.

     As far as phosphorus removal  from trickling filter plants Ts
concerned, it is recommended that further study be given to opt frot2Ing
chemical methods,  with particular attention being  given to.type of
chemical,  point of  application, and effect of recirculatlon schemes.
It is also recommended that further attention be given to the fate of
phosphorus once it is incorporated in the plant sludge, particularly
if anaerobic digesters are involved.

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                             SECTION I II

                            INTRODUCTION


      The fact that the rapid euthrophfeat ton of many lakes throughout
 this country Is one of the major water resources problems today needs no
 real convfncfng.  There Ts almost unantmous agreement among water experts
 from government, education and industry that this problem is one of  the
 most vexing ones challenging the water resources field presently and
 most probably for many years in the future.  Numerous articles which
 have appeared in the technical  journals and other publications in the
 last decade or so concur that the rate of natural  aging of many Inland
 lakes has been accelerating in recent  years,  and that this upward trend
 is the result of the increasing quantities  of "nutrients" which are
 finding their way to these waters (I)  (2) (3).   These additional  loads
 stem primarily from the varied  activities of  man residing in the water-
 shed.   Most investigators  agree that the most important "nutrients"  are
 the various inorganic forms of  nitrogen and phosphorus.   Of  the two
 elements,  the current thinking  appears  to be  that  the latter is usually
 the "limiting"  one,  meaning,  of course,  that  the extent of eutrophic
 activity is governed principally  by the concentration of  the phosphate  ion

      One principal  source  of  phosphorus to  natural water bodies  is the
 effluent of wastewater treatment  plants.  A recent study  by  Ferguson  (4),
 for example,  showed  that 44 to  56 percent of  man-generated  sources of
 phosphorus  in U.  S.  waters originate from domestic sewage.   Another
 report  (5)  prepared  by FWPCA  for  the Great  Lakes Region  states  that,
 "about  two-thfrds  of the present  annual  supply of  phosphate  going  into
 Lake Michigan (estimated to be  about 15  million  pounds) comes  from
 municipal and industrial wastewaters".

      Information of  this nature has  been  the  major impetus  in  recent
 years to develop methods for effective  phosphorus  removal  from waste-
 waters.   Early work  has been devoted to  chemical removal methods which
 have proved to be  effective but comparatively costly.  Removal of
 phosphorus  by activated sludge  treatment  has  been  shown to be possible
 at  certain  plants  and  under certain  operating conditions but there are
 still some  unanswered  questions in this area.  Very  little and  inconclu-
 sive information  is  presently available on the potentiality  for
 phosphorus  removal by  trickling filters.

     There  has been  discussion  in some quarters that  interest  fn phos-
 phorus will wane If  in fact phosphate detergent builders are replaced
 wTth other materials.  Assuming as an extreme that all the phosphorus
 is taken out of the  detergent formulations,  the concentration of phos-
 phorus  in municipal  wastewaters from human wastes alone will still be
 in the"range of 3 to 4 mg/l.  With an increasing population this amount
can continue to have a deleterious effect on receiving waters.  This  lower
value does offer a greater possibility  of removing a   large percentage
of the influent phosphorus through the  optimization of current biologl-

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cal processes alone, and without chemical additions.  This prospect Is
quite dim with the current phosphorus concentrations In wastewaters.

     Assuming solids removal Is not a problem, all biological wastewater
treatment methods depend upon one essential fact, that Is, the optimal
growth of microorganisms.  It Is through this growth that soluble organ-
Ics (or biochemical oxygen demanding materials) are Incorporated Into
part IcuI ate mass or sludge.  In addition this growth requires the
presence of nutrients, growth factors and trace salts In the wastewater
to be treated.  Two nutrients of principal concern are nitrogen and
phosphorus since limitations In either could Impair the growth of the or-
ganism Involved.  Optimum growth necessitates that a balanced quantity
of organic matter (carbon) and nutrients be available.  A wastewater
would be considered nutritionally balanced with a carbon:nitrogen:  phos-
phorus ratio of approximately 50:5:1.  This means that the actively
growing microorganism In the bio-mass of a biological treatment plant
would be extracting these elements In approximately this proportion
assuming, of course, proper environmental conditions as well as the
proper ratio of bio-mass to available food.  If the bio-mass or sludge
Is effectively removed from the system, the effluent discharged should
contain low quantities of carbonaceous matter and nutrients.

     Usually, however, the above situation Is not the case and because
of poor operation or nutritional Imbalance, nutrients are present In the
final  effluent of a biological  treatment process.  It Is not unusual,
for example, for the carbon: nitrogen: phosphorus ratio of a domestic
wastewater to be In the range of 10:5:1 which means, of course, that It
Is nutrient rich.  In such a case It Is very difficult to produce an
effluent low In nitrogen and phosphorus through biological synthesis
mechanisms alone.

     This study is concerned with the removal of phosphorus by trickling
filter slimes.  The removal,of  phosphorus by means of blo-sllmes of the
type present on a trickling >fliter medium or surface Is governed by the
same limitations stated above.   Considering the nutritional balance con-
cept there Is no question that  some amount of phosphorus will be Incor-
porated into the bio-mass following contact with the wastewater stream.
The question Is, however, can this amount be enhanced by employing  cer-
tain operational schemes?  Is there theoretically a range of biological
phosphorus uptake possibilities depending upon certain environmental or
operational conditions?  The above questions do not preclude the certain-
ty that a biological maximum uptake does exist, since It can be stated
unequivocally that It does exist.  The questions, are directed to an
elucidation of the possible enhancement of phosphorus storage In the
slime, and to a better understanding of the reasons for the variations
In reported values of phosphorus removal by trickling filter plants.  It
Is to these ends that this research project Is primarily directed.

     In a sense, the significance of this research can best be described
by considering a hypothetical community with a population of approxlmat-

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ely 1,000.  This community finds Itself In the same quagmire that many
other small communities throughout the U.S. find themselves.  Assume
that It has an existing trickling filter plant which Is common for small
communities.  The plant is up to capacity and operating as well  as can
be expected on a BOD and suspended solids basis.  It Is located In the
watershed of an Inland lake and regulatory agencies have decided with
valid reasons that phosphorus discharges must be abated or significantly
reduced.  The questions that a community In this situation first ask are
what alternatives are available?  Is chemical treatment the only answer?
Is there any way In which the existing plant can be operated which will
enhance the amount of phosphorus removal using existing plant facilities
or with minor modification or additions of equipment?  If chemical treat-
ment must be used, what is the most effective method of Its Implementa-
tion with an existing trickling filter plant?

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                         SECTION IV

                     LITERATURE REVIEW
     Recent literature reviews by Nesbitt (6),  Campbell  (7)  and  Spiegel
and Forrest (8)  on the topic of phosphorus removal  divide the present
available schemes under three main headings.   They  are chemical  methods,
biological methods and combined biological-chemical  methods.

     Of these methods, earliest attention has been  given to the  chemical
methods.  These methods Involve the use of calcium,  aluminum or  iron
salts to precipitate the phosphate ion out of solution.   The work of
Malhotra et al.  (9), Duff et al. (10), Wukasch(ll),  Leary and Ernest(l2)
and Schmid and McKlnney (13) among numerous other  works,  have  all
clearly demonstrated the effectiveness of phosphorus removal from waste-
water using this scheme.  Recent works by Ferguson  et al. (14),  (15),
have demonstrated, however, that this chemical  precipitation mechanism
is not a simple one but rather quite complex because of  the significant
effect of such variables as pH, time, carbonate and  magnesium concentra-
tion.   In addition, these studies have cast new light on the biological
vs. chemical phosphorus removal mechanism arguments  which have appeared
in the  literature in recent years.

     Following the interesting investigations of Levin and Shapiro (16),
Vacker et al. (17) and Sea If et al. (18), there has  been much Interest
in the biological methods of phosphorus removal.  Most of the recently
reported  investigations have, in addition to the three noted above,  been
confined to the activated sludge wastewater treatment process.  In all
of these  investigations it was demonstrated that under controlled oper-
ational conditions excess or "luxury" uptake of phosphorus by the acti-
vated sludge mass can be accomplished.  The rapid removal of a fixed
percentage of this phosphorus enriched sludge in the final clarlflers
results in the phosphorus removal from the wastewater.  Early studies
have concluded that the phosphorus removal activity  Is the result pri-
marily of a biochemical mechanism, whereas more recent studies (19),(20),
(21), (22), (23) suggest that other factors In addition to biochemical
ones may also be operative.

     The combined biological-chemical methods are merely the combination
of the methods already discussed.  The metal  salts  are generally added
to the mixed liquor of the activated sludge aeration basin causing both
a chemical and biological fixation of the soluble phosphate Ion. Methods
of this type have been described by Nesbitt (6), Hubbell (19), Barth
et al.  (24) and Brenner (25).

     Somewhat related to the above topics Is the question of phosphorus
release during the digestion of waste biological sludges and the Impact
of supernatant recycling on overall phosphorus removal.   Interesting
studies addressed to the question have been reported by Rtes et  al.(26),
Dunseth et al. (27), and Malhotra et al. (28).

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      A review of the literature shows rather surprisingly a serious
 dearth of Information on the effectiveness of the trickling filter
 process for phosphorus removal.  Though  one of the most often used bio-
 logical treatment processes, It Is almost totally Ignored In most re-
 ports or studies dealing with phosphorus removal.  The writer became In-
 terested In the performance of trickling filters  In this regard  after
 conducting a few phosphorus removal  surveys at several  small  trickling
 filter plants In the Milwaukee metropolitan area.  The results all  clear-
 ly showed that poor to negligible removals were being  obtained.

      The data that are available  In  the  literature seem to concur with
 the above findings.   Vacker et al.  (17)  presented some data from four
 large trickling  filter plants showing  phosphate reductions varying  from
 2  to 22 percent.   HubbelI  (19)  stated  flatly  that,  "Experience has  Indi-
 cated that the trickling  process  will  not  remove  soluble phosphate".
 Brenner (25)  and  Barth  et  al.  (29),  recently  presented  some preliminary
 results  on a  full  plant scale study  at Falrborn,  Ohio.   After a  four-
 week study where  they  dosed one-third  of the  3  mgd  plant  with  sodium alurn-
 Inate just prior  to  distribution  on  the  rock  media,  they  obtained  an
 overall  phosphorus  removal  of  65  percent.   The  phosphorus  removal across
 a  control  filter  at  the same  plant was only 15  percent.   A rather  lengthy
 report was  recently  written  by  Benson  (30)  In which  he  reported  on the
 performance of  four  trickling  filter plants  In  Wisconsin.   In  the case
 of phosphorus,  both  the literature review  and the  actual  operating data
 strongly  concur with what  already has  been  stated  above,  that  Is, phos-
 phorus  removals by trickling  filter  plants  Is generally below  20 percent.
 Finally, a  recent study by Jebens and  Boyle (31),  using a  pilot  unit,
 showed that "luxury" uptake of  phosphorus was really the  result of chem-
 ical  precipitation of phosphorus with  cations present  In the hard water
wastewater.

      Upon examination of some of the recent Interesting works by
Hartmann (32), Kornegay and Andrews  (33)  and Maler et al.  (34) on the
 kinetics of substrate removal by fixed slimes,  It  Is difficult to accept
without question that the trickling  filter process has  no potential for
exhanced phosphorus  removal.  Though these studies were concerned with
uptake of soluble carbon compounds, nutritive balance requirements and
 luxury uptake possibilities suggest higher phosphorus uptake rates than
present evidence Indicates Is possible.  Also, the similarity of  this
biota to that found  In the activated sludge process - - at  least from
the standpoint of the basic types of micro-organisms rather than  the
relative numbers of each - - further suggests the possibility of  higher
uptake rates.

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                         SECTION V

               BASIC OBJECTIVES OF THE STUDY
     The primary objective of this Investigation was to study In detail
the behavior and characteristics of fixed biological slimes from the
standpoint of phosphorus uptake.  The study was confined almost exclu-
sively to the laboratory phase of this problem, and It was not the origi-
nal Intention to Include any full scale trickling filter plant operation-
al studies relative to the question of phosphorus removal.  It was hoped,
however, that the results of this work might suggest certain operational
or design changes of the trickling filter process tn order to enhance
phosphorus removal.  The fact that soluble phosphorus might be recycled
to the head end of the treatment plant as a result of Its release Into
digester supernatant liquor was not of concern to this study.  Of course,
this would be of concern In an actual plant.  The principal objective
of this study was to examine the factors which control or have some Im-
pact on the quantity of phosphorus which Is "biochemically" Incorporated
Into fixed media slimes.  Once the phosphorus Is Incorporated Into the
slime It Is In effect removed from the soluble phase.  To keep the phos-
phorus In the slime and to remove It from the plant becomes a sludge
disposal problem.  Admittedly this latter question  Is not a simple one
and, In fact, could be one of the principal stumbling blocks In effective
biological  phosphorus removal.  At any rate, this aspect of phosphorus
removal  was not considered to be within the scope of this Investigation.
To put It very succinctly, the primary Intention of this study was to es-
tablish If certain environmental conditions or cell  growth factors cause
an Increase In the amount of phosphorus stored In the biological slime.
Could this Increase then be repeated using the same set of conditions?
Is the Initial phosphorus uptake mechanics more the result of biochem-
ical or physical adsorption activities?  Will the presence of metallic
cations to the level found In moderately hard to hard ground waters en-
hance the amount of "biological" uptake?  Will a greater percentage of
algal  growth In the biological slime result In more phosphorus storage
In the slime?  What would be the Impact of such factors as dissolved oxy-
gen,  slime thickness, nutrient balance, type of carbon source, among
others,  on the amount of stored phosphorus?  All of these questions
would appear to be of theoretical Interest only, but upon further reflec-
tion,  they all suggest certain operational  or design changes which could
be easily Incorporated Into a trickling filter process.

     In order to achieve the above stated objectives, an effective means
of growing fixed biological  slimes had to be developed.   Upon reflection,
It was decided to use a rotating disc apparatus for this purpose.  In
contrast to a full  scale trickling filter,  In the disc apparatus the
media moves past the substrate.  This difference was not felt to be Im-
portant since the main objective of the apparatus Is to grow copious
amounts  of  slime as quickly and as simply as possible.  In order to  ,
evaluate a  number of different conditions,  the unit was  constructed so
that four completely Isolated para I lei  units could be operated at the

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same time.

     In addition to the disc apparatus, another experimental  unit was
constructed to study the mechanism of phosphorus uptake on biological
slime.   This apparatus is referred to as the "channel" unit.   Four paral-
lel channels of different lengths for growing slimes were constructed on
a movable platform.  Provisions were also Included to inactivate the bio-
logical slimes growing in the channels.  Both of these units  will be de-
scribed in greater detail further on In the report.

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                         SECTION VI

               ANALYTICAL PROCEDURES EMPLOYED
     Except for the analyses and modifications to be described below,
the latest edition of Standard Methods (35) was employed as the princi-
pal guide in conducting  all the analyses used in this investigation.
In all cases, controls and blanks were Included with all  sets of samples
which had to be analyzed.  Standard curves were checked repeatedly.   On
several random occasions during the course of the study,  new control
samples were set up and analyzed.  Also a number of different analysts
cross-checked procedures.

Total Phosphorus Analysis

     Obviously one of the most Important analyses of this study was  the
one for total phosphorus.  The term "total phosphorus" can be somewhat
misleading and frequently results are misinterpreted because of this
problem.  In this study total phosphorus analysis as applied to biologi-
cal slime samples, is the sum total of all phosphorus contained in a
weighted quantity of the material.  For most of the slime grown on the
laboratory apparatus, this phosphorus is primarily the organically bound
phosphorus.  However, it should be understood that this analysis Includes
also any phosphorus which might be In the Inorganic partlculate form.
Furthermore, because the slime units were fed with a substrate containing
soluble orthophosphate (and  in a few cases soluble complex phosphate)  it
Is conceivable that this analysis Includes some of these forms also,
assuming some of the liquid film adheres to the slime surface.  Prior to
analysis, however, slime surfaces were routinely washed with distilled
water to remove as much of the soluble ortho-phosphates as possible.

     At the time that this Investigation was initiated, a number of
methods were being used for total phosphorus analysis, but none of these
were considered satisfactory in releasing completely the organically
bound phosphorus.  Recently, some methods have been developed using
strong oxldants to liberate the organically bound phosphorus.  One such
method, the "persulfate method," has shown promise due to Its simplicity
and superiority over other oxidants.  However, there was still much vari-
ation in the literature regarding the procedure which should be employed.
The "ashing method" has been considered a complete oxidation method  but
It Is a very time consuming method.  A comparison study of these two
methods was conducted to evaluate them In depth for the use In water and
wastewater analyses.

     For method development, an organic compound of phosphorus (B-glycer-
ophosphorlc acid—dlsodlum salt) was chosen to prepare a standard solu-
tion such that I ml = 0.001 mg P.

     In the persulfate method, a number of variables were optimized  such
as (a) amount of ICSJDg, (b) amount of 30JK H2SO., (c) boiling time,  and


                             II

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(d) neutralization technique prior to blue color development for color-
Imetric test.

     In the ashing method, the variables examined were (a)  amount of
MgCI *6H20 (13.67? solution), (b)  ashing temperature, (c)  ashing time,
(d) amount of 30$ I-LSO., and (e)  time of heating for hydrolysis.

     After developing these two methods for total phosphorus analysis,  a
method evaluation was conducted by analyzing the various water and waste-
water samples listed below which range from liquid to solid samples:
MlIwaukee river water, Menomonee river water,  MIIwaukee raw sewage,
Milwaukee secondary effluent, biological slime, primary sludge, secondary
sludge from the Waukesha trickling filter plant, mixed liquor suspended
solids, return activated sludge,  and vacuum filter cake.  While the com-
plete details of this study are presented elsewhere (36),  the general
findings are presented below:

          (a) The persulfate method Is capable of analyzing up to 10 mg
              of total P In the sample without dilution.  In other words,
              concentrated samples can be treated first and the dilution
              can be made afterwards.  Similar findings were obtained for
              the ashing method up to 20 mg P.

          (b) The persulfate method gave very satisfactory recovery over
              a wide range of samples, within  ±5^; except for the samples
              of mixed liquor suspended solids, return sludge, and filter
              cake.  The poor recovery with the latter samples Is probab-
              ly due to the Interference of Iron present In high concen-
              trations.  The ashing method appeared to overcome this
              shortcoming.

          (c) The ashing method proved to be superior for sludges and
              semi-solid samples and worked as well as the persulfate
              method for liquid samples,

          (d) Due to simplicity,  ease, and quickness of the persulfate
              method, this method should find wide application for the
              routine analysis of water and wastewater samples.

     Following the above comparative studies,  plus additional ancillary
studies concerning the neutral IzatIon step and hydrolysis  time which were
reported In an earlier progress report (37), It was decided that the
"ashing method" for total phosphorus analysis  would be the most accurate
for this particular Investigation.  The final  procedure employed Is as
follows:

     The slime was dried In a weighed crucible at I03°C for 8 hours.
After drying, It was returned to a desslcator and cooled for 8 hours and
weighed again.  Following weighing, 2 ml of I3.67J6 MgClj were pipetted
Into the crucible containing the dried sample.  The mixture was evaporated
to dryness at I03°C, then ashed at 600°C for 2 hours.  After cooling,


                             12

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3 ml 3056 H7SO. were pipetted into the crucible and the sample was hydro-
lyzed on a steam table at 95°C for not less than 45 rain., but not more
than 60 mln.  Immediately following hydrolysis, the sample was quantita-
tively transferred to a 200 ml volumetric flask and diluted to mark with
deionlzed water.  The contents of the flask were thoroughly shaken, and
a 2 ml aliquot (or volume to give a %T not less than 20 nor greater than
80) was transferred to a nessler tube.  The sample was diluted with de-
ionized water, I drop of phenophthalein indicator was added, and 5N
NaOH was added, drop by drop, shaking after each addition, until the
first pink color remained after shaking.  The sample was then diluted to
mark (100 ml) with delonized water.  Four ml  ammonium molybdate reagent
(See Ref. 35) and 0.5 ml (10 drops) stannous chloride reagent were added,
with thorough shaking after each addition.  After 10 minutes, but before
12 minutes, using the same specific time interval for all determinations,
the color was photometrically measured on a Beckman D.B. Spectrophoto-
meter using a I cm light-path at a wavelength of 690 mu and compared to
a standard calibration curve.  A blank was also set up using deionlzed
water plus all the same reagents used In the sample.  The phosphorus
content was recorded at a %P on a total dry weight basis.

     Standard calibration curves have been made for orthophosphate using
KhLPO. as a standard, complex phosphorus using Na,-P,0|0 as a standard,
ana organic phosphorus using B-glycerophosphorlc acid as a standard.

KJeldahl Nitrogen Analysis

     The Kjeldahl nitrogen test following the procedure In Standard Meth-
ods. (35) was first run on a standard glutamlc acid solution (.2gN/L) to
determine recovery and reproduclbiIIty.  Due to the nature of the samples
to be used In this study (solids rather than liquids), a modification had
to be made as to the form of the sample used.  In this study, the sample
was dried at  I03°C for 8 hours, cooled In a dessicator for 8 hours, and
weighed.  It was then quantitatively transferred to a Kjeldahl flask with
300 ml of delonized water.  Since It Is Important that all the dried
sample be transferred, It was necessary to carefully scrape the sample
from the crucible with a glass stirring rod.   From this point on the
analysis followed the digestion and distillation procedure as described
in Standard Methods (35) and thus both the nitrogen In the ammonia and
organic forms were Included.  The results were expressed as #sl on a dry
weight basis as follows:

          ,(NIns|llne= intrant) (0.028)
                               dry weight

assuming, of course, that the normality of the titrant was equal  to 0.02
and the dry weight was expressed In grams.

Chemical Oxygen Demand                                •

     The chemical oxygen demand analysis according to Standard Methods(35)
was first run on a standard glucose solution (.4 mgC/ml)  to determine
                             13

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recovery and reproduclb! 1 Ity.  Again, due to the nature of the samples,
a modification had to be made In the procedure.  Due to the large
amount of oxld Izab I e material present In the samples, It was not possible
to use all of the dry weight.  It was decided to wash and scrape the
dried and weighed sample Into a Waring Blender with delonlzed water and
to thoroughly blend for 10 minutes, washing down the sides of the con-
tainer several times with  delonlzed water.  The liquid was then quanti-
tatively transferred to a  500 ml volumetric flask and diluted to mark
with delonlzed water.  After thorough shaking, a suitable aliquot (usual-
ly 10 ml for actual  trickling filter slimes and 20 ml for disc slimes)
was taken and transferred  to a COD flask.  If  less than 20 ml  of sample
was used, the volume was brought to 20 ml with delonlzed water.  From
this point on, the analysis followed the procedure outlined In Standard
Methods (55).

     Initially,  the results were recorded only as - ^ — • — r—r-r — , but
after a short time,  It was decided to express them 88 rn"^ Kas?s of #C
on a dry weight basis by assuming that the reaction taking place In the
exertion of the oxygen demand Is mainly due to the oxidation of carbon-
aceous matter to carbon dioxide.  Considering the ratio of the molecular
weights of carbon to oxygen In the formation of carbon dioxide, the %C
was estimated from the COD data as follows:

             -      mg COD       3    |OQ
                                    x I0°
                mg dry weight

The validity of the above assumption was checked by comparing the %C as
determined by the COD procedure against #C as determined by a total
carbon analysis conducted In another laboratory.  This comparison was
made on three separate occasions using both slimes from the laboratory
disc apparatus and from a full scale trickling filter.  In all  cases the
slime were dried and divided Into two portions, one for each of the two
5te analyses.  The results are presented below:

          Sample #1 - Disc Slime                             % Carbon

            COD procedure, Duplicate Analysis                   45.1
                                                                47.8

            Total carbon analyzer, Private Lab.                 47.9

          Sample #2 - Waukesha Trickling Filter

            COD procedure                                   .    48.8

            Total carbon analyzer, Private Lab.                 41.3

          Sample #3 - Disc SI Ime

            COD procedure (Triplicate Analyses)                 42.6
                                                                42.0
                                                                41.3

                             14

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          Sample #3 - Disc Slime (cont.                 % Carbon

            Total carbon analyzer, Private Lab.           43,7

As noted, except for the case of the actual trickling filter slime, the
two procedures compared very well.  The disparity In the actual  slime
results may have been due to Interfering materials present In the slime.
At any rate, the results were felt to be close enough to continue ex-
pressing the COD analysis results In terms of Jfc In the slime on a total
dry weight basis.

Metal IIc Ions

     Of early Interest In the study was the development of a reliable
procedure for the analysis of calcium, magnesium, Iron and total hardness
In the slime.  Because of numerous Interfering substances in the slime,
particularly actual trickling filter slime, these procedures offered
many more problems than had been anticipated.

     First much attention was given to evaluating different digestion
procedures; namely, using a mixture of nitric and perchloric acids, and
using the dry ashing method suggested by Menar and Jenkins (38).  In the
case of the calcium and total hardness analysis, the samples were titra-
ted with EDTA following the digestion step.  Poor endpolnts In the tltra-
tlon step stimulated a search for an Improved Indicator or for inhibitors
of Interfering substances.  The details of these ancillary studies were
presented In an earlier progress report (37).  With the help of  a paper
which appeared In Ana Iy11caI  Chem i stry (39), the following tltratlon pro-
cedures were fInally establI shed

        (I)  TotaI Hardness

             a.  Pipette an amount of sample Into a porcelain dish and
                 dilute with deIonized water to a volume of 50 ml.

             b.  Add 5-6 ml of buffer and stir; then add 0.25 gm of NaCN
                 and stir;

                   (the added buffer Is needed to offset the effects
                    of  the hydrolysis of CN  In water.  The ph must
                    be adjusted to about 10 to effectively perform
                    the tltratlon)

             c.  Add 4 drops of ErI chrome Black T Indicator solution to
                 the dish.                ,  :

             d.  Add 0.005M EDTA tltrant slowly, with continuous stirring,
                 until  the last reddish tinge disappears.  The endpolnt
                 Is taken as. that point at which the solution turns to a
                 definite blue color.
                             15

-------
             e.   Calculate  total  hardness  as mg/IIter of CaCO, as
                 follows:

                    hardness  .   
-------
        c.  Ash at 800°C In a muffle furnace for one hour.

        d,  Add 5 ml of 2N HCI and digest on a steam table  for one hour.

        e.  Transfer quantitatively to 100 ml  volumetric flask and add
            10 ml of Lanthanum Oxide solution (\% La, 5% HCI)  and I  ml
            of potassium chloride solution (100 mg K per ml).

        f.  Dilute to 100 ml  mark with delonlzed water.

        g.  Analyze sample on the atomic absorption unit according to In-
            struction manual  and obtain magnitude from an appropriate
            standard curve.

        h.  Express Ca or Mg In terms of % of dry weight of slime.

     The original method used to analyze for Iron In the digested slime
sample was basically the phenanthrolIne method of Standard  Methods (35).
Interferences caused rather erratic results and extracting  the Iron with
Iso-propyl ether proved to be burdensome and the results still question-
able.  The acquisition of the atomic absorption unit proved to be the
final solution of this problem.
                               17

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                             SECTION VI I

            ANALYSIS OF ACTUAL TRICKLING FILTER SLIMES
      In order to obtain some background data on biological slimes, it was
 decided to  run phosphorus, nitrogen and COD analyses on actual trickling
 filter plant slimes.  The slime was scraped from the surface rock media
 of six municipal plants in the Milwaukee area.  All of the plants are
 within 25 miles of the City of Mi'lwaukee,  Four of the plants treat flows
 of less than one MGD.  These plants are  located in Cedarburg, Germantown,
 Hales Corners and Saukville.  The Menomonee Falls plant handles a flow
 of approximately one MGD, whereas the Waukesha plant handles an average
 flow of 10  MGD.

     After  the slime sample was brought to the laboratory, the slime
 sample was  stirred thoroughly before taking aliquots for analysis.  Usually
 the slime had a very dark green color, generally it had an earthy odor, and
 occasionally it was teeming with small (1/4 inch) pink sludge worms.  The
 si fine was analyzed in triplicate for volatile sol fds, total phosphorus,
 nitrogen and COD.  As discussed previously the mg of COD per mg of dry
 weight was  converted to percent carbon for convenience of expression.
 The slimes  were analyzed at vari'ous times to determine if marked varia-
 tions occur with seasons of the year.

 ResuIts on  Fresh TrIck Iing F i Iter SIf mes

     A summary of the slime analyses is  presented in Table  I (Appendix).
 The results are expressed in terms of percent of total  dry weight of
 sample.  The values did not vary markedly from plant to plant.  The vola-
 tile solids were mostly in the 70 to 80 percent range,  phosphorus in the
 2 to 3 percent range, nitrogen i'n the 6 to 8 percent range and carbon in
 the 40 to 50 percent range.

 Slime Washing Procedure

     Originally the method for washing the slime was to add deionlzed
 water to the sample, mix well,  and filter with paper on a Buchner funnel
 using suction.   This washing procedure was carrfed out 3 times.  There
 were several drawbacks to this method and they made it undesirable.  Even
 using coarse filter paper, the pores quickly became clogged.  To remove
 the slime from the filter paper,  It was necessary to scrape it off and
 there was the possibility of also scraping paper Into the sample.

     The second method tried, and eventually used, was centrifugatlon.
 Deionized water was added to the sample and mixed well.  The liquor was
 then put Into bottles and centrifuged at 3000 RPM (or approximately 2000 g)
 for approximately 20 minutes.  After this, the supernatant was drawn off
 and discarded.   Fresh deionized water was added to the bottle, and the
 entire procedure was then repeated.  In all, three washings were made
on the si!me.
                                  18

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Effect of Storage on Phosphorus in the Slime

     There has been some evidence published In the technical literature
that the storage of bio-mass under low or zero dissolved oxygen condi-
tions results  in the release of organically bound phosphorus to the
surrounding  liquid environment.  Most of this reported work has been
done with activated sludges.  There Is also evidence that If the phos-
phorus Is tied up in an inorganic complex, the release Is markedly re-
duced.

     It was  felt of interest to examine this behavior in actual trickling
filter slimes.  After storing the slime for a designated period of time
in a sealed  container, a sample of the sludge was taken for analysis and
washed In the manner already described to remove the "solublIized" phos-
phorus.  Low oxygen level in the bulk of the stored sludge was evidenced
by the strong septic odors and the darkening of the sludge with time.
The volatile solids also decreased with time of storage.

     The results of one storage test is presented in Table 2 (Appendix)
in which the slime sample was stored in a refrigerator (approximately
4°C).  Thus  biological activity was reduced and any phosphorus changes in
the bio-mass could be attributed to this condition.  As noted, however,
the stored phosphorus remained constant during the 15 day period.  Also
there was no difference between the initial untreated slime sample and
the initial washed sample, indicating that all the phosphorus  is held
tenaciously within the slime matrix or held by adsorpttve forces to the
siIme exterior.

     The results of room temperature storage on phosphorus release is pre-
sented in Table 3 (Appendix) using slimes from three different plants.
The results were no different from that of the refrigerated samples. The
fact that the phosphorus percentage increased with time of storage is the
result of the decrease In volatile solids with time.

     The results of 35°C storage on phosphorus percentages In  the sludge
is presented in Table 4 (Appendix).  As noted, the results are not marked-
ly different from those obtained under room temperature conditions.  Also
there was little difference in the behavior of the slimes from the differ-
ent plants.

Effect of Anaerobic Storage on Phosphorus In the Slime

     The preliminary results on the effect of storage on phosphorus
levels In actual  trickling filter slimes were of interest and  prompted
further studies along these lines.  It was decided to store the slime
under truly anaerobic conditions and at an elevated temperature to speed
up the rate of biological  activity.  Approximately 100 ml  of trickling
filter slime was placed In a 500 ml Erlenmeyer flask and the remainder
of the flask was filled with deionized water.  The flask was tightly
sealed with a water trap device so that generated gases could  be re-
leased.   A very small  void space was provided between the surface of the


                             19

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supernatant liquid and the bottom of the stopper.   Samples of super-
natant were taken at various times and analyzed for ortho-phosphate.
Deionized water was added to the flask to compensate for the sampling
loss.  A number of flasks were set up in order to make it possible to
periodically analyze the slime Itself during the degradation process.
When this was done, the flask content was discarded and the phosphorus
was then monitored in a new flask.

     The results of two of these studies are presented in Tables 5 and 6
(Appendix).  As noted In both test runs, the supernatant phosphorus built
up to a maximum value of approximately 200 mg/l within four days and re-
mained fairly constant beyond this time up to 26 days of anaerobic stor-
age.  There Is an apparent Inconsistency In the fact that as the super-
natant phosphorus level  increased, the percentage phosphorus value in
the slime remained the same or even increased.  As noted previously, the
phosphorus percent is calculated on a total dry weight basis and the de-
struction in volatile solids tends to mask the overall decrease in the
phosphorus present in the solid fraction.  Considering the data in Table
6 (Appendix) and expressing the percentages of phosphorus In terms of
the solids in the slime, shows that this value was approximately 16 per-
cent at the start of incubation and 10 percent 25 days later.

     The same approach described above was attempted using a composite
slime from the four discs of the laboratory apparatus.  The results of
this test are presented in Table 7 (Appendix).  This test run was beset
with more problems than was the case with the actual trickling filter
slimes.  The laboratory slime was very light and fluffy and tended to
rise up to the supernatant liquid during the anaerobic storage period or
when disturbed slightly In the process of obtaining the sample.  As a
consequence the data obtained were somewhat erratic.  However, it is evi-
dent that there was release of phosphorus to the supernatant.  Better than
50 percent of the phosphorus remained in (or with) the slime even after
13 days, which Is particularly surprising since this phosphorus was
supposedly metabo11caI Iy incorporated into the slime.

     Finally, three additional anaerobic storage studies were conducted,
two using actual trickling filter slimes and one using laboratory disc
slime, but with the difference that the percent carbon and nitrogen In
the slime were also monitored.  The results of these studies are present-
ed in Tables 8, 9 and 10 (Appendix).

     As noted In all three cases, an immediate supernatant build-up In
phosphorus occurs within 5 to 7 days and remains fairly constant from
that time on.  It could be reasoned that In the case of the actual trick-
ling filter slimes the phosphorus released Immediately was from organic
sources while that held in the slimes was  Inorganically bound.  Such
reasoning does not fully explain the results obtained with the laboratory
slime since It is expected that almost all the slime phosphorus is organ-
ically bound.

     As would be expected, the percent carbon  In the  incubated slime


                             20

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samples also decreased with time, which means of course, the organic
carbon was converted to methane and carbon dioxide as a result of an-
aerobic decomposition.  The reduction in percent nitrogen indicates that
either some of the organic nitrogen In the slime was solubalized or con-
verted to nitrogen gas.  These results merely attest to the active an-
aerobic activity which took place In the vessels during the storage of
the siime samples.

Other Studies

     The most likely reason for the rather limited release of phosphorus
under storage conditions is that the phosphorus In the slime is bound up
into some sort of inorganic complex.  The metallic ions of calcium, mag-
nesium and iron, among others, would be involved in this complexing ac-
tivity.  It is therefore of interest to have some knowledge of the "total
hardness" of the actual trickling filter slimes used In the storage ex-
periments.  The procedure for hardness determination has been described
earlier In the report.  The hardness values found In some of the slimes
analyzed are shown in Table II (Appendix).  The results are expressed in
terms of percent as calcium carbonate.  The much lower values for the lab-
oratory slime are expected since the feed solution was made with softened
water.

     The high hardness values of the slime are not surprising when one
analyzes the hardness in the effluents of some of the treatment plants
used as sources of the slime.  The average hardness and calcium of trip-
licate analyses of three plant effluents expressed as calcium carbonate
are presented below.

     Plant            Calcium (mg/l)           Hardness (mq/l)

     Hales Corners        210                          439

     Menomonee Fa 11s       300                          593

     Waukesha             201                           388
                             21

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                        SECTION VIII

    DESIGN AND CONSTRUCTION OF LABORATORY DISC APPARATUS
     Past experience of the writer with slime growth studies, demonstra-
ted that it would be possible to use a rotating disc device to grow the
slime required for the phosphorus uptake investigation.  Whereas in a con-
ventional trickling filter plant the wastewater is sprayed over a fixed
rock media, it was felt that for a laboratory set-up, it would be much
more convenient to move the media through the wastewater or test sub-
strate.  Also, in contrast to an actual trickling filter plant, the basic
purpose of the laboratory disc apparatus Is to provide a convenient means
of developing copious growths of biological slimes on a solid surface,
rather than using this surface activity for the BOD reduction in the In-
fluent stream.

     A schematic sketch and photographs of the laboratory disc apparatus
are presented in Figures I  and 2 respectively.  The apparatus was con-
structed so that it would be possible to run four test runs in parallel
at the same time.  The reaction vessels and 10 Inch discs were construct-
ed of one-half inch plexiglas.  The apparatus was constructed so that It
would be possible to run the entire unit under one set of conditions by
removing two plugs provided in each of the three interior separation
walls.  Each reaction vessel was fed by gravity from 19 liter jugs con-
taining the feed solution.   The siphon arrangement was set up so that the
inlet atmosphere air pressure point was approximately one inch above the
point of the siphon discharge line.  This made it possible to obtain a
fairly uniform rate of discharge since the driving head remained constant.
The final adjustment of the discharge rate was made using a pitch-cock on
the rubber tubing used for the discharge line.  An air gap was provided
between the discharge line and the influent funnel  of the reaction vessel
to prevent "crawling" contamination of the feed solution with biological
slime.  The discs were rotated by means of a pulley system connected to
a gear reduction unit and an electrical motor.  The exact rotational
speed was accomplished using a rheostat-type unit controlling current to
the motor.

     After passing through  the reaction vessel and past the rotating disc,
the feed solution passed out from the bottom of the unit into a settling
cone (plastic Imhoff cones  were used for this purpose).   The level  of the
liquid in the reaction vessel  and thus the degree of disc submergence was
controlled by adjusting the height of the sett I Ing cones.   The supporting
structure for the cones was constructed to easily facilitate this adjust-
ment.  Most of the slime which sloughs off the discs was collected in the
bottom portion of the cones which were provided with a volumetric scale
for estimating sludge quantities.  The effluent from the cones was finally
directed to large storage jugs.   The entire system operated on gravity
and, except for the disc rotation, did not depend on mechanical  equipment
for Its operation.


                            22

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  STOPPER
                        FEED LINE
                                          SCHEMATIC OF  LABORATORY

                                                 DISC  APPARATUS
                       PINCH
                       COCK
                                 10" DISC
                              FOR  SLIME GROWTH
x X f •' ' S f/ S ' /ff/S

        AIR GAP2-"
       // s '

          REACTOR
            VESSEL
   MOTOR  AND
SPEED CONTROL
        7
                                                      SETTLING
                                                        CONE
                                            \  •
                                                      xSLUDGE
                                                      STORAGE
                                                  EFFLUENT
                                                   JUG
                                                               S S S / / / /
                                                             FIGURE !

-------

r r r »-\f "
       •
n^^^r^^^^^^~
   FIGURE 2


PHOTOGRAPHS OF

DISC APPARATUS
                  (a) overalI view  of
                      apparatus
                  (b) close up view of  disc
                            with si I me
 24

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                         SECTION IX

       TESTS CONDUCTED WITH LABORATORY DISC APPARATUS


General Test Procedure

     At the start of a test run the disc vessels were seeded with biolo-
gical slime obtained from a full scale trickling filter plant.  The four
reaction vessels were then fed a prescribed substrate at the rate of
approximately 9 liters per day.  The feed carboys were filled with 18
liters of substrate every two days.  The feed rates were continually ad-
justed during this period so that approximately 9 liters of substrate
were used up In 24 hours.  No attempt was made, nor was It felt necessary,
to keep a constant flow rate throughout the entire period.

     As the feeding progressed the disc surfaces were frequently examined
for evidence of slime build-up.  Usually within two to three days a fair-
ly good growth had developed.  However, In most cases the units were fed
for approximately two weeks before slime samples were obtained for analy-
sis.   In a few cases the test runs were shorter and longer than this
period, but In all cases the runs were continued to the point where it
was felt that the slime growth was completely acclimated to the particular
test run conditions.

     Just prior to the filling of the feed with fresh substrate, the jugs
and siphon assembly were thoroughly washed in a strong bleach solution
and carefully rinsed with tap water and distilled water.  The air inlet
line to the feed Jug was plugged with sterile cotton.  All the feed jugs
were washed with a strong acid solution at the end of each test run.

     Each day the walls of the reaction vessels were thoroughly brushed
to prevent bull'd-up of any slime.  As a result, slime build-up.on the
walls was minimal.  Agitating the vessel contents periodically provided
good "seeding" for the disc surfaces.  During the course of each run, the
pH, temperature and dissolved oxygen were determined In the reaction
vessels.  A careful log was kept of all activities associated with the
test run.  Such Items as date of Jug refill, disc speed check,.condition
of disc slime, sludge In storage cone, etc. were noted In the log book.

     After sufficient time had elapsed for good slime growth development,
the disc rotation was stopped and the top half of the slime covered disc
was gently washed ,with delonlzed water.  The slime was then scraped from
the surface of the disc with a steel knife blade and placed Into a beaker.
After completing the top half of the disc, the lower half was exposed and
the slime was removed In the same manner.  The analyses of the slime were
set up Immediately after the four scrapings were made.  A residual coat-
Ing, or film, remained on the disc and served as a seed for the subsequent
test run.  The analyses routinely conducted on thesiIme were vojatlle
sol Ids, phosphorus, nitrogen and COD.  In a few test runs calcium and
                             25

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magnesium analyses were also conducted.  On an average of once per test
run, the same analyses were conducted on the feed solution and effluent
of the disc units.  This  was done to provide a random check of the char-
acteristics of the feed solution and to have a record of the "removals"
provided by the disc units.  As expected, very limited nitrogen and phos-
phorus removal was obtained, particularly in the runs where excess amounts
of these elements were in the substrate.  The COD removals were usually
around 50 percent.  These results were not surprising since this apparatus
was designed to grow slime rather than to treat an influent flow.

     Attempts were made to measure the slime thickness with a callper de-
vice and a magnifying glass, just prior to the scraping of the slime.  Be-
cause of irregularity and the "spongy" nature of the slime the measure-
ments obtained were approximate at best.  After the thickness was noted,
the scraped slime contained on each disc was placed into a calibrated
beaker and the volume of "crop" was also noted.  Other characteristics
of the slime,such as color and texture, were also noted and recorded in
the log book.

Specific Test Runs

     As pointed out previously, one of the principal  objectives of this
investigation was to determine under what set of conditions It was
possible to enhance the amount of phosphorus storage in biological slime.
At the outset, no attention was given to applicability of the condition
to a prototype treatment system employing fixed biological slimes for the
treatment of wastewaters.  The question was simply: Can greater amounts
of phosphorus be taken up into biological slimes, and If so, under what
environmental and operating conditions?  Once this is established, the
second question logically follows:  Is there any way  of Incorporating
these same conditions In a prototype system to achieve enhanced removals
without going to well-known chemical removal methods?  The subsequent dis-
cussion on the test runs conducted should be viewed with the above objec-
tives In mind.

     For the first test runs It was decided to feed the four disc reactors
with a specified amount of carbon, nitrogen and phosphorus.  The carbon
and nitrogen were kept identical  but the phosphorus values were varied
from a phosphorus deficient feed In the case of the first disc reactor,
to a phosphorus rich feed in the case of the last.  The nutrient composi-
tion on a weight ratio  basis selected for the four feeds was as follows:

                            C       N         P

             Feed I          50      5         O.I
             Feed II         50      5         |
             Feed III        50      5         5
             Feed IV        50      5        10

Feed II  would be considered  the closest to a nutritionally balanced  diet
                             26

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from the standpoint of the metabolic requirements of most nvfcroblal  sys-
tems, Including biological slimes.

     For the first series of test runs It was decided to use glucose as
the carbon source.  The sources of  nitrogen and phosphorus were ammonium
chloride and sodium phosphate, respectively.  The concentration selected
for the glucose was 300 mg/l which  results In a theoretical  COD of 320
mg/l for the feed solutions.  With  this concentration established, the
resulting concentrations of the nitrogen and phosphorus would have to be
as follows:
                                       Concentration (mg/l)
                                     GIucose      N        P

            Feed I                     300
            Feed II                    300
            Feed III                   300
            Feed IV                    300

The glucose and the ammonium chloride were added to 80 liters of softened
Milwaukee tap water In a  large plastic container and the contents were
stirred thoroughly.  The phosphate  salt was added directly to the four
Jugs.  The softened tap water containing the glucose and nitrogen sources
was then pumped up to the four feed jugs.  A disc speed was selected and
maintained for the entire duration  of the test run.

     Following this Initial series  of test runs, new operating conditions
were employed in subsequent runs.  A summary log of all test run conditi-
ons, Including some remarks on s.llme characteristics, Is present In
Table 12 (Appendix).  As noted, the operating variables Include disc
speed, water hardness, type of carbon source, type of nitrogen source,
concentration of phosphorus In the  feed, absence or addition of a calcium
source to the substrate, and whether or not the disc slime was continual-
ly subjected to a plant light.

      In the nomenclature  used to Identify each test run, a different
roman numeral was  used for each carbon source; "A" means the first test
run under a set of conditions and "B" means the second test run under
the same set of conditions, "H" means hard water was used to make up the
feed solution and  "S" means soft water was used to make up the feed solu-
tion, and the  last number represents the disc speed In revolutions per
minute.                                        ,

     The term "hard" water  Is really a relative one since It Is Milwaukee
tap water which Is obtained from Lake Michigan.  The hardness of this
water Is approximately 120 mg/l as CaCO, which Is only moderately hard
for a water supply.  The soft water was obtained by passing Milwaukee
tap water through  a zeolite softener.  The.hardness of this water for
all practical purposes is close to zero.                  ^         -

     The carbon sources used were glucose, nutrient broth, yeast extract
and dry milk solids In various combinations.  These same substrates
                             27

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served as the nitrogen source in addition to ammonium chloride.  Sodium
phosphate, an ortho-phosphate, was used as the phosphorus source in all
cases and concentrations ranging from 0 to 24 mg/l as P were employed.
Two sources of calcium were used in those test runs in which this element
was added, namely, calcium chloride and lime.

     For most of the test runs the disc siIme was subjected to the normal
diurnal light cycle.  The disc apparatus was located In a room with a
southern exposure and thus well lighted during the day, In spite of the
fact that the shades were down on bright sunny days.  On many days the
fluorescent ceiling lights were on continually.   In spite of this light-
ing, only a limited amount of green growth developed on the discs.  For
some of the final test runs it was decided to subject the disc apparatus
to 24 hours of special plant lighting* with a wave length for optimal
photosynthetic activity, to determine the effect of copious algal growth
on phosphorus storage.  From the luxurious green growth which developed
during these runs, it was apparent good algal growth had occurred.

Results of Tests Conducted With Laboratory Disc Apparatus

     The results of all of the test runs described in the previous section
are presented in Table 13 (Appendix).  The four slime analyses Included
in this Table are volatile solids, phosphorus,  carbon and nitrogen.
Calcium and magnesium analyses were also conducted in some of the test
runs, and these results are presented in Table 14 (Appendix).  Because
varying amounts of phosphorus were fed to the four reaction vessels, the
results are presented for each of the individual  disc slimes.  Because of
the incompleteness of the data in a number of test runs, not all of the
test run data were Included in the statistical  analyses which will be de-
scribed subsequently.  The test runs included are designated in the Table
and as noted, 33 of the 38 runs are included.

Other Operating Observations

     A detailed log was kept of the observations  made during the operation
of the disc apparatus.  Early In the investigation tt was felt that since
a biological process was being operated, these  observations might assist
in the final interpretations of the data.   While such items as slime
color, texture and yield may not appear too significant for an individual
test run, they might supply some important Insight if certain finds were
noted In a large number of test runs.  For example, runs with high biologi-
cal phosphorus uptake might be associated  with  a  slime of a particular
character.  If this proved to be the case, this slime could then be ex-
amined in  greater detail.

     After a number of test runs It became apparent that no dfscernable
trends were developing In this regard.  On the  contrary, the results were
 ^General Electric, Plant Light, F40PL (two lamps)


                             28

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often conflicting and confusing and thus can be only used in a general
qualitative way rather than  in a specific quantitative or statistical
way.

     For most of the test runs the slime had a color in the white-cream-
yellow range.  Occasionally the slime also had a pink tint but this never
persisted.  Except for the test runs in which the continuous plant light
was employed, very little green growth developed on the disc surface.
This was surprising since the apparatus was located in a well lighted
room.  Copius green colored slime developed during the last test runs
when the apparatus was subjected to the plant light.  In some of these
test runs part of the surface was also covered with a more tan-1 ike
growth.  In summary, if one examines all of the test run data on slime
appearance, no real conclusions can be drawn other than possibly the fact
that slime growth is a highly hetereogeneous biological  situation.

     The same conclusions can be drawn regarding slime texture.  At times
the slime was very slimey and fragile and had the tendency to break off
the disc surface in large chunks.  Other times It was much more cohesive.
The former condition was more prevalent at slower disc speeds whereas
the latter was more apt to be the case at higher speeds.   Sometimes this
variation occurred in the four discs which were all rotating at the same
speed but under varying feed conditions.  When the feed solutions con-
tained some hardness, the slime surface on occasions had a more granular
texture - - particularly at higher disc speed.

     Much attention was given to slime thickness.   One of the original
premises of this investigation was that slime thickness might have  an
effect on the amount of phosphorus stored in the biological  slime.   Speed
control  of the disc rotation was provided mainly In an attempt to control
slime thickness.  A call per device was employed for some of  the early
test runs to measure slime thickness.   This proved to be difficult  be-
cause of the fragile nature and Irregularity of the slime surface.   A
micrometer was tried for subsequent test runs and  this offered somewhat
of an Improvement.   The micrometer was operated under a large magnifying
glass mounted on the reaction vessel.   The major problem again was  the
Irregularity of the surface.  Frequently the slime was quite thin where
large chunks of slime had previously fallen off.   Thus there were fre-
quent areas of thin and thick slime on the same disc.

     In general, the slime thickness was greatest  for the'slower test
runs, namely test runs of 25 RPM disc speed and less.  These slimes were
usually in the  1/8 to 3/16 inch range of thickness.  At higher speeds
the slimes were generally thinner; namely in the 1/16 to possibly 1/8
Inch range.  Any attempt to refine these data beyond the above generali-
zation would be quite futile.  This Is particularly true when one con-
siders that the slime thickness often varied from one disc to another in
the same test run in which all four discs operated at the identical speed.

     It Is to be expected that as thickness varied so did slime yield.
By slime yield Is meant the total amount of slime "crop" removed from the
                                     %                 >

                             29

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surface of the disc at the end of a test run.  Again more crop was usual-
ly obtained for the test runs at the slower speed.  Though variations
occurred above and below these amounts, the amount of yield was generally
In the 75 to 100 ml range with a highly variable amount of actual  total
solids.  Obviously slime yield is not the same as the total sludge gen-
erated during a test run.  No attempt was made to arrive at this latter
figure.  Slime yield represented the amount of slime carried on disc sur-
face under a given set of operating conditions after achieving a steady-
state condition with the substrate In the reaction vessel.

     At various times during the course of the Investigation the temper-
ature, pH and dissolved oxygen content of the substrate In the reaction
vessel were measured.  Most of the temperatures were In the 18 to 20°C
range and the pH In the 7.0 to 8.0 range.  The dissolved oxygen varied
depending on the disc speed of the particular test run.  At no time did
the D.O. ever reach zero.  Even for the test runs of I  RPM disc speed, *
the D.O. Was generally In the 1.0 to 2.0 mg/l range.  At disc speeds of
10 to 25 RPM the D.O. Increased to the 4.0 to 6.0 mg/l  range,  and at
higher speeds a D.O. of 7.0 to 9.0 mg/l was achieved.
                            30

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                         SECTION X

  DESIGN AND CONSTRUCTION OF LABORATORY CHANNEL APPARATUS
     One question which has been raised regarding the uptake of phosphor-
us by biological  slime is the role that physical  adsorption on the slime
surface plays in the overall  removal process.   It Is known that this
mechanism is very important in natural  soils.   As far as phosphorus re-
moval ts concerned, it makes  little difference if the phosphorus is In-
corporated biologically into the slime or adheres to the slime surface
as long as It is removed from the wastewater.   If, however, it is deter-
mined that surface adsorption is an Important part of the overall removal
mechanism, the total amount of slime surface area In a fixed medium treat-
ment device would partly control the extent of the total phosphorus which
is removed.  This is a design feature which could be controlled to some
extent by the designer.

     A channel apparatus was  constructed to study this particular ques-
tion.  A schematic of the apparatus set up Is  presented in Figure 3.
Most of the channel unit was  constructed of 1/2" plexiglas.  An important
design feature was that the slope of the channel  plank could be adjusted
as desired.  The greater the  slope, the less will be the time of contact
between the slime surface and the phosphorus containing feed solution.
Also important was the fact that the four different channel lengths were
available on the channel plank.  At a given slope, this provided still
another variable, that is, total.opportunity for slime contact.  A photo-
graph of the channel apparatus Is presented In Figure 4.

     The channels themselves  were one-half Inch wide and one-quarter Inch
deep.  The channels were constructed so that It was possible to firmly
lay down on the bottom a fine fI berg I as mesh.   The mesh provided an ex-
cellent base for the development of the channel slime.

     Four positive displacement pumps which operated off the same motor
drive were used to bring the  feed solution to the head end of the channel.
The pumps maintained the desired low rate of flow for extended periods of
time with very little variation among the four units.  After dropping to
the channel surface, the feed solution flowed  down the channel to a dis-
charge hole at the end of the reach.  Plastic tubing was used to drain
the channel effluent into sample jugs.

     Brackets were provided at the ends of the channel plank to support
an ultraviolet lamp assembly  used to Inactivate the microorganism on the
channel surface.  The  lamp assembly did not interfere with the Influent
line of the channel operation.

     A special metal tool was designed to scrape the slime growth (or
crop) from the channel surface.  The scraping  edge of the tool was de-
signed such that It lifted the slime off of the bottom mesh.
                             31

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SUPPLY. LINE FOR
EACH  CHANNEL
         SCHEMATIC    OF   LABORATORY
              CHANNEL  APPARATUS
                                                FOUR-  1/2  WIDE CHANNELS
                                                   FOR  GROWTH OF SLIME
                                                     ( LENGTHS SHOWN)
                            ADJUSTA
                             PLANK
MOTOR  AND
   PUMPS
EFFLUENT
  LINES
                                                  ADJUSTMENT
                                                     ARM
          120 LITER
          FEED  CONTAINER
                                    SUPPORT FOR
                                    UV LIGHT UNIT
                                    £/  //  //  /  /////////
                                     L-EFFLUENT  JUGS
                                                                        FIGURE   3

-------
       FIGURE 4
PHOTOGRAPH OF CHANNEL




      APPARATUS
         33

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                          SECTION XI

      TESTS CONDUCTED WITH LABORATORY CHANNEL APPARATUS


General Test Procedure

      After the channel apparatus was constructed, the first problem was
to establish the operating conditions that resulted in a good slime
growth in the four channels.  The first substrate tried was dry milk
solids in varying concentrations but the resulting growth was not too
satisfactory.  The slime growth was very Irregular and "puffy" in tex-
ture.  Various rates of flow were also tried.  When the flow rates were
too  low (in the range of I  to 2 ml per minute per channel) the slime
growth tended to be Irregular and not cover the entire channel surface
area.  A flow rate of 4.0 ml per minute per channel eliminated this prob-
lem.  The logistics of supplying enough feed substrate in the continuous
operation of a laboratory unit precluded Increasing the flow rate too
much above the 4.0 ml figure cited above.

      Glucose was also tried as the carbon source, supplemented with ammo-
nium and phosphate salts.  The characteristics of the slime did not Im-
prove much.  A combination of nutrient broth and glucose was also tried
with limited success.  After attempting a number of different combina-
tions, the substrate which appeared to work the best consisted of glucose,
milk solids, sodium phosphate and ammonium chloride added to softened
Milwaukee tap water.  The actual amounts of each of these varied during
some of the initial test runs, but for the most of the remaining test
runs the channel  feed substrate consisted of the following:

           80 liters of Milwaukee softened tap water
           25 g.  dry milk solids (commercial brand)
           75 g.  glucose
            15 ml  Na^HPO. solution (containing 20 mg P per ml)
           80 ml  NH^ CI solution (containing 48 mg N per ml)

A typical analysis of the above feed solution was as follows:

           COD = 1330 mg/l
     Kjeldahl N =   65 mg/l
       Total-P =  5.5 mg/l

      One of the factors which also had a bearing on the^characteristics
of the final substrate selected was the sensitivity of phosphorus removal
within the  length of channels available in the apparatus.  In some of the
original test runs, negligible removals were obtained.  There were three
reasons for this situation.  First, metabolic requirements for phosphorus
are quite low relative to carbon and nitrogen and thus little phosphorus
will  be Incorporated  into the channel slime even under Ideal growing
conditions.  Second, in some of the initial feed solutions the carbon to
phosphorus ratio was too low.  Greater phosphorus removal sensitivity can
                              34

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be obtained by using a carbon rich - phosphorus poor nutritional composi-
tion In the feed.  Thirdly, the length of the channels themselves afford-
ed a limited amount of "reaction" surface for biological  phosphorus re-
moval.  The substrate composition described above appeared to minimize to
a certain extent the effect of the first two conditions.   Not much could
be done with the channel length condition since this is governed by the
normal physical  limitations of a laboratory set-up.

      Three different slopes were used during the course of the channel
studies.  They were as follows:

                                    Tangent of
          Slope Designation         Slope Angle           Angle

              Low                     0.0616               3.5°
              Mid                     0.4412              24.0°
              High                    0.9537              43.5°

The procedure for starting a test run was essentially the same In all
cases.  After the feed solution was made up in a 80 liter batch, the solu-
tion was seeded with a small amount of slime from a full  scale trickling
filter plant.  Some of the seed material was also rubbed Into the fine
bottom mesh for the entire  length of the channels.  After a growth started
to develop, the channels were continually fed with fresh  feed solution.
After a good growth of slime developed, It was scraped down to the level
of the fine mesh.  Test runs were normally conducted several  hours after
the slime was scraped.  Effluent samples for analysis were collected In
erlenmeyer flasks packed in Ice.  Prior to analysis, the effluent and  raw
samples were filtered through paper.  The procedures employed for the
analysis of the liquid and slime samples were the same as those used for
disc studies.

Channel Studies Using Ultraviolet Radiation

       In order to establish the degree of  physical  adsorption  involved  In
the  uptake  of  phosphorus by biological  slimes, a method  of Inactivating
the  surface slime had to be developed.  The method  used  could  not alter
the  surface properties  of  the  slime appreciably since  It was necessary
that the physical .characteristics of the slime surface be the  same both
before and  after the biological  Inactlvatlon.  Only then would  It be
possible to establish whether  the physical adsorption mechanism was Im-
portant  in  phosphorus uptake.

       Several  methods of siIme surface Inactlvatlon were considered.  For
example, a  solution containing a chemical  disinfectant such as chlorine
or  Iodine could  be passed  down the channel for a short period of: ,tlme
just prior  to  resuning  the tesfv  Another  method could be to place the
entire channel set-up In the walk-In constant temperature room.  The
channel; apparatus was designed-with tKlsposslbllIty  in mind.  The tem-
perature of  the  room could be  reduced['to about 5°C and the slime accli-
mated  to this  temperature.  At this temperature the biological activity


                               35

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 would be suppressed while the physical  characteristics would remain  un-
 altered.  Still  another possibility was one reported in a paper by
 Borchardt and Azad (21) in which chemicals like sodium azide,  sodium ar-
 senate, iodoacetic acid, and sodium fluoride could be used for the  i nac-
 tivation of phosphorylating enzymes while not harming the cells them-
 selves.  Finally, a set-up could be constructed containing a bank of
 ultraviolet germicidal  lamps which could be employed to radiate the
 channel slime.  Keeping in mind  the main idea of stopping the  biochemical
 activity of the  slime while still  retaining the same physical  character-
 istics of the surface,  this last method appeared to offer the  greatest
 promise.  Thus a number of ancillary studies were conducted to explore
 i ts potent i a I.

       In the first ancillary test some  slime was removed  from  the disc
 apparatus and  divided into two petri  dishes.   One was retained as the
 control, the other was  subject to UV light in a commercial  unit used  for
 the disinfection of glassware needed for bacteriological  tests.  After
 exposing the slime for  an  increment of  time,  a piece of sterile gauze was
 placed gently  on the slime and transferred to tryptone  glucose extract
 agar plates.   The same  procedure was used for the control.   Ten exposures
 with a total cumulative time of  120 minutes  were used.  After  incubation
 the plates  were  compared.   The UV  plates had  fewer colonies than the  con-
 trol,  but the  kill  did  not appear  to be extensive enough.   One reason for
 the poor kill  might have been that an irregular (not like  a surface grow-
 ing slime)  slime sample was  used and  the UV  radiation could not reach all
 the surface  because of  projections.

       An  undisturbed  slime sample,  1-1/2  inch  by  1-1/2  inch, was carefully
 removed  from the  disc for  the  second test.  The test  was conducted as
 described above,  only that  180 minutes of total UV exposure was employed
 this time.  Some  kill was  noted  at  10 minutes.  At 20 minutes the kill
 was  good, and at  30  minutes  it  was  very good.   After 60 minutes there were
 very few colonies on  the plate incubated with  the gauze from the UV
 radiated siime.

       Another test  was  conducted using a slime sample which was felt to be
 essentially undisturbed.  This was accomplished by attaching a small
 piece  of plastic  film to the  rotating discs.   After two weeks the slime
 growth on the film  was  continuous with the slime on the remainder of the
 disc surface.  On the day of the test the small film was carefully re-
 moved  from the disc with the slime completely  intact.  The  results ob-
 tained with this slime  sample were about the same as those described
 above.

       Except for some minor  problems, the results of these ancillary tests
 were felt to be encouraging and  it appeared that this method could be
 used to  inactivate  the  slime.  One problem noted was that  in the commer-
 cial unit the temperature  in the enclosure began to  increase with time
 of exposure.   In  all of the tests the internal temperature at the point
 of slime radiation  increased from 24°C to 38°C.  Such an increase In
 temperature could be  deleterious  to the slime surface.  The increase in
 temperature also  caused a drying  of the slime surface which In turn
could change the  physical characteristics.  When sterile water was placed
on the slime surface after a period of UV exposure,  It caused an Increase


                                   36

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in the number of colonies of the UV plates.   The water seemed to re-
suspend or disturb the surface colonies.   This is not surprising since
it is known .that UV penetration ib very minimal, particularly in an
aqueous environment.  It was felt that the first problem could be mini-
mized by designing the UV light assembly so  that the heat dissipated
rapidly.  The second problem could be minimized by continuing the UV
radiation of the slime for long periods while the feed solution was  ac-
tually flowing down the channels.

      The UV light assembly was constructed  to completely cover the  chan-
nel planks which contained the four channels.  The depth of the unit was
eight inches with the result that the lamps  were approximately three
inches from the slime surface.  Four 36 inch long, commercial germicidal
lamps* were mounted in the unit such that all the channel surface was
well  covered.  The top of the unit was constructed of thin gauge sheet
metal for good heat dissipation.  The sides  of the units were constructed
of standard window panes.  Large openings were provided at the two ends
of the unit (influent end and discharge ends of the channels) in order to
insure good air circulation.  The unit was constructed so that it was
well  secured to the channel plank and in no  way interfered with the
slope adjustments or the channel flow arrangement.

Specific Test Runs

      As stated previously, a large number of preliminary test runs  were
conducted in order to establish the most suitable substrate composition
for development of the best channel slime.  During the test runs, analyses
were conducted on the channel feed and effluent.  The analyses conducted
were total phosphorus, COD and total Kjeldahl nitrogen.  During these
test runs, analyses were also conducted on the channel slime.  These con-
sisted of percent total phosphorus in all cases, plus percent nitro-
gen,  volatile solids and carbon in that order depending upon the amount
of channel slime available.

      Following the establishment of a suitable substrate, a series  of test
runs  were conducted primarily to determine If the amount of phosphorus
uptake varied with the length of channel  slime.  These tests were con-
ducted at three different slopes to appraise the importance of time of
contact on the amount of phosphorus uptake.

      As noted previously, one of the main reasons for the channel study
was to determine if physical adsorption is part of the mechanism of  phos-
phorus uptake on biological slimes.  At this point the UV light assembly
was incorporated into the test runs.  Two test runs were first conducted
as follows.  The channel  unit was fed with the substrate until a good
slime growth developed.  Following this development, Influent and effluent
analyses were conducted as before.  The channel unit was then subjected
to UV radiation as the channels were fed in  the normal manner.  Influent-
effluent analyses were conducted following varying increments of UV  ex-
posure.  The longest time of exposure employed was 72 hours.

*General Electric, Germicidal, G30T8

                                   37

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      Following these tests a number of "starve-ki11" test runs were
conducted.  The basic idea in these runs was to first grow an active slime
and then run a conventional influent-effluent test as described above.
The slime was then "fed" with de-chlorinated tap water for 24 hours which
kept the slime viable but also In a somewhat starved condition.  The chan-
nel unit was then fed with the normal  substrate while a second test was
being conducted.  The water feed was resumed for 24 hours,  the UV radia-
tion was turned on for two hours with  the water feed continuing.  At the end
of the radiation period, the normal feed was again turned on and in-
fluent-effluent analyses were conducted.  This procedure was employed at
the three different slopes.

Results of Tests Conducted With Laboratory Channel App_ar_atus_

      The analysis of channel slimes during preliminary runs using a num-
ber of different substrated Is presented in Table 15 (Appendix).

      The results of feed and effluent analyses for eight test runs is
presented in Table 16 (Appendix).  It  wasn't until test run VI that the
test substrate was finally established for the remainder of the channel
testing program.  Slime analyses conducted during this phase of the
testing program are presented In Table 17 (Appendix).

      Feed and effluent analyses while the UV assembly was in use are
presented in Table 18 (Appendix) and the results of the "starve-ki 11"
runs are presented in Table 19 (Appendix).
                                 38

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                            SECTION XI I

                       DISCUSSION OF RESULTS
     The prfmary objective of thi's study was to determine if the amount
of phosphorus uptake by biological sli'mes on fixed surfaces can be
enhanced.  To accomplish the objective i't was necessary to examine how
vari'ous operational  variables, such as, carbon source,  concentration of
Influent phosphorus, dissolved oxygen level, slime thickness, water
hardness, and degree of algal growth, Influenced the amount of phos-
phorus stored i'n the si ime.   ft was also important to understand the
mechanism of phosphorus uptake on biological slimes, particularly the
importance of physical  adsorption on the slime surface.

     It was never the  intention of this research project to develop a
"new process" for fixing phosphorus on biological  slimes; but rather,
It was to develop a better understanding of this uptake activity, and
on the basis of this understanding, establish if it would be possible
to suggest ways to enhance the amount of phosphorus removed  in the
conventional trickling filter process.  An enhancement of 15 to 25
percent, if accomplished through relatively simple operational or
design changes, would be quite significant.  This would be particularly
true In the event that the phosphorus additives are eventually elimi-
nated from detergent products.  This would  lower the amount of total
phosphorus  in raw waste-water from approximately 10 mg/l to 3-4 mg/l.
Well controlled biological treatment might adequately take care of these
lower concentrations eliminating the need for more costly chemical
treatment.  Even at these  lower concentrations it would be erroneous to
assume that biological  treatment will ever provide "total" phosphorus
removaI.

     The research program was mainly centered around two  laboratory
apparatus which already have been described in detail, namely, the disc
apparatus and the channel apparatus.  The discussion of the  results for
convenience will also be divided under the same two headings,

The Disc Apparatus

     In spite of the large number of test runs and the various opera-
tional  conditions used, the percentage phosphorus on a dry weight basis
stored in the disc slime ranged from O.I I to 4.34 percent, as noted  in
Table 13.  Discounting the ten lowest values and the ten highest values,
the range decreases markedly to 0.30 to 2.88%, which is a way of demon-
strating the central tendency of the data.  The range  is quite narrow
considering that well over 400 different disc slime samples are  included,
By different samples Is meant each  is from a new growth on a given disc.
It is true that many of the test runs were repeated with  identical oper-
ating conditions, but each test run resulted in a completely new growth.
As noted previously, all si fine'was scraped from the disc prior to the
start of a new run.  .In spite of the same operating conditions for two
different test runs, frequently the slime had a different appearance and
consistency.  It would be  reasonable to suspect that such differences
would be the result of the growth of varying biota which  in  turn might
result in a change In the amount of phosphorus stored.
                                 39

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Thus for the purpose of discussion, each slime sample can be considered
as being derived from a unique situation,

      A cursory observation of Table  13 will show that a vast majority
of the percent phosphorus values in the slime vary from 1.5  to 2.5 per-
cent.  Values considerably below 1.5 percent occurred primarily In those
test runs where the concentration of the phosphorus in the feed was quite
low - - In fact a level which would be considered biologically deficient.
In these test runs the ratio of carbon to phosphorus was approximately
500 to I.  Under these feed conditions there appears to be enough phos-
phorus to cause growth but certainly not enough to satisfy the full de-
mand of the growing siIme.

      On the other hand, values above 2,5 per cent usually occurred In
those test runs where some calcium hardness was added to the feed.  In
general, the greater the amount of hardness In the feed, the higher is
the resulting phosphorus stored in the slime.  Obviously these higher
phosphorus values are not the result of biological activity but the
result of calcium phosphate precipitates being incorporated into the
slime matrix.  This fact  is substantiated by the values for percent cal-
cium in the slime, which were also greater for the slime samples in which
the phosphorus  amounts were greater.

      It  is much more difficult to explain the reasoning for the varia-
tion In stored phosphorus for the  range between  1.5 to 2.5 percent.  An
examination of Table  13 wlII not show any clear trends or patterns.  If
anything, the results appear haphazard and random.  When one thinks he
has an explanation for the results of a particular run, the hypothesis
is quickly refuted in a subsequent run.

      In  an attempt to derive some meaning from the spate of data aval  I-
able from the disc apparatus, it was decided to subject the data to a
multiple  correlation analysis using a digital computer.  The method em-
ployed Is based on the assumption of  linearity In the relationships
among the variables.  The purpose of this statistical analysis Is to
establish which of the  independent variables have the greatest signi-
ficance  in controlling the eventual percent phosphorus stored In the
slime.  The data employed as Independent variables Include disc speed,
concentration of COD, N,  P and Ca  In the feed, and percent volatile
solids, C and N In the disc slime, whereas, the percent P In the slime
was naturally the dependent variable.  The results of the multiple re-
gression  analysis for the data from each of the four discs Is presented
in Tables 20, 21, 22 and  23.  A summary of two common statistics used
to characterize multiple  regressions  Is presented In Table 24.

      The results of this statistical analysis are by no means conclusive,
However,  keeping In mind  the constraint that  linearity between variables
was the assumption in all cases, the  results suggested the following
observations:

      (I)  The quantity of calcium  In the feed Is the most Important
           variable influencing the quantity of stored phosphorus.

                                 40

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                                        TABLE 20

                      MULTIPLE LINEAR REGRESSION ANALYSIS - DISC

                                        (a)
                        ALL  TEST  RUNS
RUNS WITHOUT CALCIUM ADDED
                           (b)
VARIABLE
Speed (RPM)
Feed COD( mg/l)
Feed N (mg/l)
Slime VS .'('*)
Slime C (*)
Slime N (*)
Feed Pi mg/l)
Feed Ca (mg/l)
Slime P <50
Mean
41.7
347.1
17.9
90.4
48.4
8.5
3.8
43.7
1.33
Stand
Dev.
38.9
81.2
8.1
4.2
9.7
2.2
3.4
54.9
0.62
Corr.
Coef.
0.25
0.27
0.28
-0.33
-0.01
0.77
0.69
0.49

"t" Value (c) Mean
Case A
1.58
0.07
0.31
I.II
0.81
5.08(d)
6.23(d)
5.,2
-------
M
                                                 TABLE 21

                               MULTIPLE LINEAR REGRESSION ANALYSIS - DISC II

                                                .(a)
                                 ALL  TEST  RUNS
                                              RUNS WITHOUT  CALCIUM  ADDED
                                                                        (b)
VARIABLE
Speed (RPM)
Feed COD(mg/t)
Feed N(mg/l)
Slime VS (%)
Slime C (2)
Slime N (%)
Feed P (mg/l)
Feed Ca(mg/l)
Mean
41.7
347.1
17.9
89.7
47.6
9.1
6.3
47.1
Stand
Dev.
38.9
81.2
8.1
2.8
5.3
I.I
3.0
54.1
Corr.
Coef .
-0.05
0.32
0.21
-0.38
-0.04
0.03
0.31
0.44
"t" Val
Case A
l.98(d)
l.66(d)
0.09
2.42(d)
,.68(d)
1 .03
0.55
1.59
(c)
ue
Case B


2.l7(d)
l.96(d)


0.23
l.84(d)
Mean
38.2
353.8
19.3
91.3
48.3
9.2
5.12

Stand
Dev.
37.3
107.8
10.5
2.34
6.52
1.3
2.7

Corr.
Coef.
-0.25
0.53
0.47
0.19
0.15
0.32
0.09

"t" Value(c)
Case A Case B
l.89(d)
-.33 0.57
0.44 0.65
0.76
0.47
0.91
0.62 1.26

    Slime P
    (Dep. Var.)
1.73  0.42
1.54    0.39
                      (a)  33 test runs  included
                      (b)   19 test runs  included
                      (c)  Each case for the  independent variables shown
                      (d)  Significant to 90$  level of confidence

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                                       TABLE  22

                    MULTIPLE  LINEAR  REGRESSION  ANALYSIS  -  DISC TTT

                      ALL  TEST   RUNS             RUNS  WITHOUT  CALCIUM  ADDED
VARIABLE	

Speed (RPM)    41.7

Feed CODCmg/I)347.I

Feed N(mg/|)   17.9

               89.4

               47.3

                9.3
SIime VS ($)

SI line C <*)

SIi me N (%)
Feed P (mg/l)  10.4
Stand Corn.
Dev. Coef.
38.9
81 .2
8.1
3.4
4.5
1.0
2.1
54.8
0. 12
-0.02
-0.12
-0.58
-0.31
-0.26
-0. 12
0.73
it
t" Val
Case A
0
0
0
1
0
0
0
2
.30
.77
.50
(d)
.91
.43
.94
.48
.84d>
(c)
ue
Case B


0.72
(d)
1 .69


0.00
j.ei"
Mean
38.2
353.8
19.3
91.1
48.8
9.7
10.5

Stand
Dev.
37
107
10
2
4
0
2

.3
.8
.6
.8
.9
.9
.5

Corr.
Coef.
-0.
0.
0.
0.
0.
0.
-0.

18
II
05
31
33
59
15

(c)
"t11 Value
Case A
(d)
1 .97
0. 17
1.00
0.52
(d)
1 .85
(d)
0.44

Case B

0.61
0.46


0.53

SI i me P (%)
(dep. Var.)
                1.82  0.44
1.55   0.25
               (a) 33 test runs included
               (b) 19 test runs included
               (c) Each case for the independent variable shown
               (d) Significant to the 9056 level of confidence

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                                            TABLE 23
                         MULTIPLE LINEAR REGRESSION ANALYSIS - DISC  IV

                                        .(a)
                         ALL  TEST  RUNS'
                                            RUNS WITHOUT CALCIUM ADDED
                                                                       (b)
Mean
VARIABLE
Speed
Feed
Feed
SI ime
Slime
Slime
Feed
(RPM)
COD(mg/l)
N (mg/l)
VS (?)
C (%)
N (%)
P (mg/l)
Feed Ca (mg/l)
41
347
17
88
46
9
15
55
.7
.1
.9
.5
.2
.2
.6
.4
Stand
Dev.
38.9
81 .2
8.1
3.9
4.5
1.2
4.9
58.4
Corr.
Coef.
0.13
-0.08
-0.17
-0.85
-0.25
-0.39
-0.19
0.78
H
t" Value(c)
Mean
Case A Case B
0
0
0
5
0
0
2
3
.32
.88
.06 l/9|(d)
.I6(d)5.84(d)
.48
.28
..l(d)2.72(d)
.30(d)4.07(d)
38.2
353.8
19.3
90.8
46.7
9.5
17.3

Stand
Dev.
37.3
107.8
10.6
2.2
4.9
1.3
5.9

Corr.
Coef.
-0.32
-0.05
-0.09
-0.34
0.22
0.04
0.17

"t" Value(c>
Case A Case B
0.21
0.25 0.25
0.36 0.29
1.18
0.85
0.98
1.34 0.56

Slime P (%)
(Dep.Var.)
1.96    0.60
1.59    0.22
                 (a)  33 Test runs included
                 (b)  19 Test runs included
                 (c) Each case for the independent variable shown
                 (d) Significant to the 90% level  of confidence

-------
                            TABLE 24

              SUMMARY OF MULTIPLE CORRELATION AND F
                 VALUES 3ASED ON MULTIPLE LINEAR
                   REGRESSION ANALYSIS OF DATA
                       FROM DISC APPARATUS


MULTIPLE
DISC INDEPENDENT VARIABLES INCLUDED CORRELATION
1 Al 1
1 N,
1 Al 1
1 COD
II All
II N ,
M All
1 1 COD
II 1 All
III N,
III All
1 1 1 COD
IV All
IV N,
IV . All
IV COD

P & Ca in feed & % MS in slime
except Ca in feed
, N & P in feed

P & Ca in feed & % VS in s 1 i me
except Ca in feed
, N & P in feed

P & Ca in feed & % VS in s M me
except Ca in feed
, N 4 P in feed

P & Ca In feed & % VS in slime
except Ca in feed
, N & P in feed
0.954
0.893
0.988
0.846
0.741
0.615
0.730
0.600
0.773
0.759
0.837
0.222
0.921
0.917
0.573
0. 188
it p ti
VALUE
30.46
27.69
64.04
12.65
3.66
4.27*
1 .80*
2.82*
4.47
9.50
3.68
0.26*
16.82
37.18
0.77*
0. 18*
*These values are not significant at 95% level of confidence

                               45

-------
           Insuf f i'cfent data are available to show if there is any
           difference as to the calcium source.

      (2)  Usually, as the percent volatile solids in the slime increases,
           the quantity of stored phosphorus decreases.   This is the same
           as saying that as the amount of inorganic calcium phosphate precipi-
           tate increases in the slime, the percent volatile solids decreases.

      (3)  The results are much more random and  inconclusive for the runs
           in which calcium was not added to the feed.  This indicates
           that the biological  phenomonen is in  a sense more random and
           unpredictable even under the control  conditions of the labora-
           tory.  The possibility of running a biological  unit with the
           basic aim of controlling the quantity of phosphorus in the
           slime appears quite remote.

      (4)  It appears that the best situation in which it might be pos-
           sible to predict or control  biologically stored phosphorus
           is when the amount of feed phosphorus is limiting.  In other
           words, feed phosphorus is a  much more sensitive parameter when
           it is biologically limiting.   If one  compares the mean stored
           phosphorus in runs without calcium addition for the four discs,
           the values for discs II, III, and IV  are essentially identical
           and below the values for disc  I.  It  should be recalled that
           the quantity of phosphorus in the feed to the four discs was
           varied, with only disc I being phosphorus limiting.

      (5)  On the basis of the calculated multiple correlation co-efficients
           and "F" values, it would be  safe to state that the percent
           phosphorus expected in the slime can  be predicted from knowing
           the values of all  the independent variables employed, but again
           it should be stressed that the quantity of calcium in the feed
           tends to mask all  other variables.

      (6)  Disc speed, the only variable that characterizes a physical
           feature of the phosphorus uptake process, appears to be a
           significant factor about one half of  the time.   At the start
           of the research project, the suggestion was raised that as the
           disc speed increases and the resulting slime thickness de-
           creases, the quantity of phosphorus stored per unit weight
           would tend to increase.  By  virtue of the fact that both posi-
           tive and negative correlation co-efficients were obtained for
           the various disc data, one would be forced to reject this
           hypothesis outright.

      One of the main objectives followed throughout the course of this
investigation was to vary operational conditions and to determine what
effect these changes had on the quantity of phosphorus stored in the bio-
logical  slime.  It was hoped that one or two set conditions could be
                                  46

-------
found which encouraged a biota that tended toward a "luxury" uptake of
phosphorus.  Once this condi'tion or these conditions were identified,
an  in depth study of them would then follow, including bacteriological
analyses, to understand the mechanism of what was going on.  Ideally,
this understanding could be applied to prototype treatment processes
employing bio-slimes on fixed surfaces.  For this reason slime was grown
under light and dark conditions, with different carbon and nitrogen
sources, under varying disc speeds, etc.  However, the stored phosphorus
never consistently increased above the predictable 1.5 to 2.5 percent
range unless it was "forced" with hardness additions.  A biological
"arrangement" was being sought after which would provide "luxury" phos-
phorus values in the 3.5 to 5.0 percent range.  This could then lead one
to a means of enhancing the amount of phosphorus removed by treatment
processes using fixed biological slimes.  Regretably, this "arrangement"
was not found or does not  exist.

The Channel Apparatus

      As pointed out previously, much of the time devoted to the channel
apparatus was spent trying to develop a slime growth that could be effec-
tively used for the investigation.  This problem turned out to be much
more troublesome than had been anticipated.  Once a slime was developed,
the analysis of the key constituents of the slime demonstrated that the
channel  slime growth was similar to the disc growth.  For example, the
percent volatile solid of the channel slime for some preliminary runs
varied from approximately 86 to 91, the percent phosphorus from 1.2 to
2.2 and the percent nitrogen from 8.1 to 10.7.  Later, more extensive
analysis of the channel slime produced similar results.  These values
are similar to the ones obtained for a typical disc slime.  Thus it can
be assumed that the nature and make-up of the two slimes were fairly
similar — a conclusion which would be suggested by a visual comparison
of the two siImes as well.

      After establishing a set of operating conditions which resulted in
a good channel  slime growth, a number of simple feed-effluent experiments
were conducted.  Runs were set up merely to demonstrate that a measure-
able removal of phosphorus can take place within the length of channel
slimes and slopes provided.  The only concern at this point was to show
that some activity (certainly some of which had to be biological since
it is known that phosphorus is incorporated Into the living slime matrix)
was operative in causing the removal of phosphorus.  It was also demon-
strated at this time that COD and N were being removed from the feed
solution as it passed down the channel slime.  The concurrent removal of
P, COD and N certainly supports the obvious conclusion that biological
activity of the growing slime was mainly responsible for the difference
in the feed and effluent constituent concentrations.  What was not ob-
vious at this point, however, Is whether or not all of the phosphorus
removed from the feed solution was needed for metabolIc reasons.

      As expected, generally the amounts of P, COD and N removed varied
in a direct proportion to the length of the channel.  Effluent concentra-
                                 47

-------
 tions  In almost all cases were  lower for the  longest channel.  The
 difference was particularly striking when comparing the shortest and
 the  longest channels  (18  inches vs 72  inches).  Again, though this was
 expected,  It  is still not possible to  establish if the difference was
 entirely due  to a biological mechanism, at  least  in the case of phos-
 phorus  removal.

      The change in the slope of the channel  did  not appear to have a
 significant effect on the overall results.   If a  comparison is made
 between the "high" slope and the "low" slope  results, it appears that
 the effluent  concentration for  the latter condition was  lower, though
 thse results  are not  too conclusive by any means.  In other words,
 time of contact was not a significant  variable regardless of the
 mechanism which is operative in causing the  uptake of phosphorus.  This
 may not be unreasonable if one  considers that the channel flow condi-
 tion was estimated to be  laminar even  under  the highest slope condition.
 Using the form of the Reynolds  number  as defined  by Bird et al. (40), a
 typical flow  rate of  4 ml per minute,  and the geometry of the slime
 channel, a Reynolds number of 21 was estimated.   It Is suggested by the
 above  reference that  "laminar flow without  rlppltng" occurs in a channel
 when the value is between 4 and 25.  The value of the Reynolds number
 would, of course, be  considerably lower under the two flatter slop flow
 conditions.   Under laminar flow conditions the kinetics of passage of
 phosphorus through the water film is governed by  molecular diffusion.   In
 order  for molecular diffusion to occur a concentration gradient must set
 up  in the water film  directly above the slime interface.  The uptake of
 phosphorus at the interface will result in the formation of this gradient,
 but  it  is of  little or no consequence whether the gradient results from a
 biological or physical mechanism.  Thus the  fact  that time of contact was
 not a significant variable does not give any  Indication which mechanism is
 actually operative.  All  It indicates  Is that under the three slope con-
 ditions employed, phosphorus was taken into  the slime at the maximum rate.
 Had turbulent conditions prevailed, It Is likely  that the uptake kinetics wou |
 would have been Increased since the maximum  possible concentration of
 phosphorus is always  present at the sllme-lfquld  interface.  This is par-
 ticularly true if the physical  mechanism Is  significant.

       It was  not until the ultraviolet radiation  studies were completed
 that a clearer insight as to the uptake mechanism was obtained.  A
 series of test runs were conducted in which  P, COD and N analyses
 were compared for the feed and  effluent from the  channel apparatus, both
 prior to and  following ultraviolet radiation  of the slime surface.
 Under all three slope conditions a significant removal of P, COD and N
 occurred prior to the ultraviolet exposure.   Following exposure to the
.ultraviolet radiation, removals dropped to  zero.  Exposure duration was
 varied from  I to 72 hours with  no appreciable change In results.  The
 characteristics of the slime surface did not  appear to be altered by
 the radiation procedure.  Apparently the radiation technique was effec-
 tive in attenuating the biological activity  at the surface of the slime
 since both the COD and N remained unchanged  following passage of the
 feed solution down the channel.  Such was not the case for all the pre-
                                   48

-------
vlous test runs.  The fact that there was no phosphorus uptake during
these same runs means that this activity is strictly the result of a
biological mechanism.  There was no evidence of any phosphorus uptake
resulting from a physical adsorption on the slime surface.

     A second series of test runs was set up to test this thesis fur-
ther.  In this series an initial control run was conducted  at the
three difficult slopes without any radiation of the slime surface.
The results, as before, showed a measureable amount of phosphorus uptake
In all cases; the amount being proportional to the length of the chan-
nel.  The slime surface was then "fed" for 24 hours with dechlorinated
tap water which resulted in a starved situation.  The surface was fed
the test substrate following this starvation period and the results on
the effluent were obtained as before.  Because of the starved condi-
tion, It was theorized that the biological  uptake of the phosphorus on
the slime surface would be intensified.  While a good uptake was ob-
tained in all cases, it was not necessarily an improvement  over the in-
itial or control condition, thus, starvation did not markedly enhance
the amount of biological phosphorus uptake.  Following the  test run
just described, the slime was again subjected to 24 hours of water
feed followed by two hours of ultraviolet radiation of the  surface.
This feed was continued while the slime surface was subject to the ultra-
violet treatment.  The test substrate was then fed to the slime and the
previously described analyses were conducted on the channel effluents.
As before, negligible amounts of phosphorus were removed by the channel
slime.  Some removal was noted in a few cases, but because  COD and N
were also removed, it is probable that this removal was due to partial
biological activity rather than physical adsorption.  Thus  In these
cases the ultraviolet treatment was not sufficiently effective in In-
activating the slime surface.  In some of the runs the test procedure
was repeated following 24 hours of ultraviolet exposure, but the-re-
sults were similar to those already described for the 2 hours of
exposure.
                                49

-------
                           SECTION XIII

                         ACKNOWLEDGEMENT
     The writer wishes to express his gratitude to Mr.  Edwin  F.  Barth,
Project Officer, Biological Treatment Research Program, Advanced
Waste Treatment Research Laboratory, EPA, for his interest and  co-
operation throughout the course of the investigation.

     Appreciation is also extended to Beth Hugunin for  taking care  of
the vast number of laboratory analyses in an expert fashion during
her two years with the project, to William Karlovitz for his  assistance
in the laboratory and with the statistical studies, to  Andrew Horn  for
his construction of the fine laboratory apparatus, and  to the various
Milwaukee area communities which provided trickling filter slime
samp Ies as des i red.
                                50

-------
                         SECTION XIV

                          REFERENCES
 (I)  Stewart, K.M. and Rohllch, G.A., "Eutrophlcation - A Review,"
         A Report to the State Water Quality Control  Board,
         California, 1967.

 (2)  "Eutrophlcation:  Causes, Consequences, Correctives," Proc.  of
         Symposium, Nat. Acad. of Sclen.,  Washington, D.C.,  1969,
         661  pp.

 (3)  House Report No.  91-1004, 91st Congress,  2nd Session,  Twenty-
         Third Report by the Committee on  Government  Operations,
         April 14, 1970.

 (4)  Ferguson, F.A., "A Nonmyoptlc Approach to the Problem of  Excess
         Algal Growths,"  Envlr. Scl. and  Tech., March 1968, pp.188-193.

 (5)  "Water Pollution  Problems of Lake Michigan and  Tributaries,"
         U.S. Dept. of  the Interior, FWPCA,  Great Lakes Region,
         Chicago, III., Jan. 1968.

 (6)  Nesbltt, J.B., "Phosphorus Removal—The State of the Art,"
         presented at 40th Annual Conf., Water Pollution Control
         Assoc. of Pa., Aug. 7-9, 1968.

 (7)  Campbell, George R., "Studies on the Chemistry  of Orthophosphate
         and Polyphosphate Removal with Ferric Chloride," M.S.  Thesjs,
         Rensselaer  Polytechnic Institute,  Troy, N.Y., 1967.

 (8)  Spiegel, M. and Forrest, T.H., "Phosphate Removal: Summary  of
         Papers,"  J. of the S.E.D. of ASCE, Vol. 95, No. SA5,
         Oct. 1969, p.  803.

 (9)  Malhotra, S.K., Lee, G. Fred and Rohllch, G.A., "Nutrient
         Removal  from Secondary Effluent by  Alum Flocculatlon and
         Lime Precipitation/1  Int. J. Air and  Wat. Poll.,  pp.  487-500,
         I964<                             ;   '•

(10)  Duff, J.H., Dvorln, R. and Salem, E.,  "Phosphate Removal, by
         Chemical Precipitation,"  2nd Workshop.on Phosphorus Removal,
         U.S. Dept. of  the Interior/Chicago, III ./June 26-27, 1968.

(II)  Wukasch, R.F., "The Dow Process for  Phosphorus  Removal,"  2nd
         Workshop on Phosphorus Removal-j 4J.S. Dept. of the  {nterlbr/
         Chicago, 111., June 26-27; 1968.

(12)  Leary,  R.D. and Ernest, L,A;, "Municipal  Utl ItzatIon of an
         IndustrI a I  Waste F6r Phosphorus RemovaI," 32nd Porcelain
         Enamel Institute, Univ. of III.,  Oct.  8,

                              51

-------
 (13)  Schmid, L.A. and McKinney, R.E., "Phosphate Removal by Lime-
         Biological Treatment Scheme,"  Jour. WPCF, Vol. 41, No. 7,
         1969, p.  1259.

 (14)  Ferguson, J.F. and McCarty, P.L., "The Precipitation of Phosphates
         from Fresh Waters and Wastewaters,"  Tech. Rept. No. 120,
         Stanford Univ., Dept. of Civil Engr., Dec. 1969.

 (15)  Ferguson, J.F., Jenkins, D. and Stumm, W., "Calcium Phosphate
         Precipitation in Wastewater Treatment,"  Applied Chem.  Lab.,
         Div. of Engr. and Applied Physics, Harvard Univ., Cambridge,
         Mass.

 (16)  Levin, G.V. and Shapiro, J., "Metabolic Uptake of Phosphorus by
         Wastewater Organisms,"  JWPCF, pp. 800-821, June, 1965.

 (17)  Vacker, D., Connell, C.H. and Wells, W.H., "Phosphate Removal
         Through Municipal Wastewater Treatment at San Antonio,  Texas,"
         JWPCF,  pp. 750-771, May, 1967.

 (18)  Scalf, M.R., Pfeffer, P.M., et. al., "Phosphate Removal at
         Baltimore, Maryland,"  J. of the S.E.D. of ASCE, Vol.  95, SA5,
         Oct. 1969, p. 817.

 (19)  Hubbell, George E., "Process Selection for Phosphate Removal at
         Detroit,"  presented at 41st Annual  Conf., WPCF, Sept.24,1968.

 (20)  Hennessee, T.L., Maki,K.V., and Young,  E.Y.,  "Phosphorus  Removal
         in Wastewater by a Modified Activated Sludge Process,"   City
         of Trenton, 1968.

 (21)  Borchardt, J.A. and Azad, H.S., "Biological  Extraction of
         Nutrients,"  JWPCF,  pp.  1739-1754, Oct. 1968.

 (22)  Personal Communication  from Milwaukee Sewerage Commission,
         Milwaukee, Wisconsin, 1969.

 (23)  Muibarger, M.C., Shifflett, D.G., Murphy,  M.C.,  and Huffman, D.D.
         "Phosphorus Removal  By Luxury Uptake,"   Jour. WPCF,  Vol.  43,
         No. 8,  Aug. 1971, p. 1617.

 (24)  Barth, E.F., Brenner, R.C.  and  Lewis, R.F.,  "Chemical-Biological
         Control  of Nitrogen  and  Phosphorus in Wastewater'EffIuent,"
         JWPCF,  pp. 2040-2054, Dec.  1968.

 (25)  Brenner, R.C., "Phosphorus  Removal  by Mineral  Addition,"
         Technical  Symposium—Nutrient Removal  and  Advanced  Waste
         Treatment,  Cincinnati,  Ohio, April  29-30,  1969, p.  1.9.

(26)  Ries, K.M.,  Dunseth, M.G.,  Salutsky, M.L., Shapiro, J.J.  "Ultimate
         Disposal  of Phosphate From Wastewater by  Recovery as
         Fertilizer,"  Final  Report,  FWPCA Contract No.  14-12-171,
         July 15,  1969.

                             52

-------
(27)   Dunseth,  M.G.,  Salutsky,  M.L.,  Ries,  K.M.  and  Shapiro, J.J.,
        "Ultimate Disposal  of  Phosphate  From  Wastewater  By  Recovery
        as Fertilizer,"  Report Submitted to  Fed.  Water  Poll. Cont.
        Adm.  (No. 14-12-171),  Jan.  1970.

(28)   Malhotra, S.K., ParriIlo, T.P.  and Hartenstein,  A.G.,  "Anaerobic
        Digestion of  Sludges Containing  Iron  Phosphates,"   J.S.E.D.
        of ASCE, Vol. 97, No.  SA5,  Oct.  1971,  P. 629.

(29)   Barth,  E.F., Jackson,  B.N.,  Lewis, R.F.  and  Brenner,  R.C.,
        "Phosphorus Removal  From Wastewater by Direct  Dosing of
        Aluminate to  a Trickling Filter,"   U.S.  Dept.  of the  Interior,
        Adv.  Waste Treatment Res.  Lab.,  Cine., Ohio, June  1969.

(30)   Benson, R.J., "A Report  of the  Performance of  Four Trickling
        Filter Plants In Wisconsin,"   M.S.  Thesis, Univ. of Wisconsin,
        Madison, 1970.

(3D   Jebens, H.J. and Boyle,  W.C.,  "Enhanced  Phosphorus Removal  In
        Trickling Filters," J. of  the S.E.D.  of ASCE, Vol. 98, No. SA3,
        June  1972, p. 547.

(32)   Hartmann, L., "Influence of  Turbulence  on the  Activity of
        Bacterial Slimes,"  JWPCF,  pp. 958-964,  June 1967.

(33)   Kornegay, B.H.  and Andrews,  J.F.,  "Kinetics  of Fixed-Film
        Biological Reactors,"   JWPCF,  pp. R460-R468  (Part 2),Nov.  1968.

(34)   Maier,  W.J., Behn, V.C.  and  Gates, C.D., "Simulation  of the
        Trickling Filter Process,"  J. of the S.E.D. of  ASCE, Vol. 93,
        No. SA4, pp.  91-112, Aug.  1967.

(35)   Standard Methods for the Examination  of  Waters and Wastewaters,"
        APHA, AWWA and WPCF,   13th  Edition, 1971.

(36)   Gupta,  K.B., "A Comparison of  the  Persulfate and the  Ashing
        Analyses for  the Total  Phosphorus," M.S. Thesis, Marquette
        University, Dec. 1970.

(37)   Zanoni, A.E., "Progress  Report on  Phosphorus Removal  by Trickling
        Filter Slimes,"  Grant No.  I70IODZ6,  Marquette Univ., Civil
        Eng.  Dept., Milwaukee,  June 1970.

(38)   Menar,  A.B. and Jenkins,  D.,  "The  Fate  of Phosphorus  In Sewage
        Treatment Processes,"   Part II,  SERL  Report  No.  68-6,
        University of California Berkeley,  August  1968.

(39)   Hlldebrand, G.P. and ReMley,  C.N.,  "New Indicator for Complex-
        ometrlc Titratlon of Calcium In  Presence of  Magnesium,"
        Analytical Chemistry,  Vol.  29, No.  2,  Feb. 1957, p. 258.

(40)   Bird, R.B., Stewart,  W.E. and  Lightfoot, E.N., "Transport
      Phenomena,"  J. Wiley 4  Sons,  Inc., N.Y.,  I960,  p. 41.

                            53

-------
SECTION XV




 APPENDIX
    54

-------
                                            TABLE  I

                              ANALYSIS  OF TRICKLING  FILTER  SLIMES
                             FROM SIX PLANTS IN  THE  MILWAUKEE AREA
Date

Cedarburg
Dec. 11/1969
Jan. 6, 1970
Apri I 21,  1970
April 28,  1970
Germantown
Jan. 6,  1970
Hales Corners
Nov.  II,  1969
Nov. 20.  1969
% Volati le Sol ids
Each
Sample Avg.
73.24
74,84 74.64
75.85
81 .41
82.44 82.05
82.30
80.77
80.90 80.82
80.79
80.95
80.68 80.44
79.70
69.10
75.31 70.82
68.05
79.01
82.40 78.64
74.52
81.72
80.17 81.25
81.85
% Phosphorus (b)
Each
Sample Avg.
2.62
3.56 3.03
2.92
1.99
2.04 1.99
1.95
2.54
2.38 2.46
2.47
2.32
2.63 2.52
2.61
2.28
2.40 2.26
2. 10
2.00
2. Of 2.13
2.38
2.36
2.54 2.40
2.29
% Nitrogen (b)
Each
Sample Avg.
6. 15
6.10 6.12
6.38
6.23 6.40
6.60
7.35
7.61 7.46
7.42
7.33
7.47 7.35
7.24
6.54
6.6t 6.65
6.79
7.89
7.94 6.05
2.31
7.83
7.19 7.57
7.69
% Carbon
Each
Samp le
40.14
38.89
41.42
46.46
37.50
49.61
44.85
44.92
38.21
40.35
28.50
30.86
40.35
45.90
43.16
45.49
44.36
43.76
39.75
(b)
Avg.
40. 15
41.98
46.46
39.28
33.24
44.85
42.62

-------
Date

Menomonee Fa I Is
Nov. II, 1969
Nov. 20,  1969
Dec. 22,  1969
Saukvllle
Jan. 6,  1970
April  21,  1970
 April  28,  1970
 Waukesha
 Nov.  4,  1969
 Nov.  II,  1969
                         TABLE I  (continued)

% Volatile Solids  % Phosphorus  Cb)     % Nitrogen (b)
Each               Each
Samp Ie    Avg.     Sample     Avg.
79.58
79.52
79.61

79.81
79.75
75.18

78.69
78.52
78.37
80.42
82.. 32
82.37

75.28
73.99
74.28

74.28
73.34
72.95
                               79.57
                               78.25
                               78.53
                               8E.70
                               74.52
                               73.52
3,
2.
.3,

2.
2.
2.
03
86
02

80
88
84
                   2.88
                   2.79
                   2.85
                   2.34
                   2.2f
                   2.40
2.
2.
2.

2.
2.
II
20
30

19
39
74.85
76.34
76.14
                               75.78
                                        2.29
                   3.08
                   3.02
                   3.21

                   2.75
                   2.57
                   2.65
                               2.97
                              2.84
           2.84
           2.32
                              2.20
                              2.29
                                                   3.10
           2.66
% Nitrogen (b)
Each
Sample Avg.
7.66
7.52 7.57
7.52
7.17
7.51 7.26
7.10
6.95
6.65 6.65
6.36
7.76
6.62 7.35
7.66
7.07
7.19 7.15
7.19
6.62
6.59 6.62
6.66
6.63
6.81 6.72
5.97
6.90 6.52
6.69
% Carbon
Each
Sample
36.37
36.00
36.64
43.69
43.97
43.95
41.55
39.60
35.81
45.22
47.21
33.56
43.35
41.81
46.76
37.99
34.91
41.96
46.76
41.62
46.54
35.44
40.99
39.75
Cb)
Avg.
36.34
43.87
38.99
42.00
43.97
38.29
44.97
38.73

-------
                                             TABLE I  (continued)

                    % Volatile Solids  % Phosphorus (b)     % Nitrogen (b)          % Carbon (b)
                    Each               Each                 Each                 Each
Date                Samp I e    Avg._     Samp I e     Avg.      Samp I e     Avg.      Samp I e

Nov. 20, 1969       75.21              2.05                 6.62                 43.35
                    74.53     74.93    2.62       2.47      6.75       6.70      42.22       42.80
                    75.05              2.74                 6.72                 42.82

Dec. 9, 1969        73.74              2.65                 6.05                 41.06
                    74.21     73.49    2.80       2.89      6.49       6.25      43.20       45.00
                    72.53              3.23                 6.22                 50.74

Dec. 22, 1969       71.79              2.60                 5.48                 39.26
                    73.93     72.86    2.54       2.58      3.22       4.53      41.14       40.91
                                       2.61                 4.90                 42.34

Jan. 9, 1970        72.94              2.76                 6.27                 28.16
                    72.01     72.64    2.92       2.77      6.42       €.34      46.01        39.19
                    72.97              2.64                                      43.39

Feb. 14, 1970       75.64              3.29                 6.44                 47.55
                    71.99     72.10    3.16       3.51      7.06       6.57      45.26       45.22
                    68.68              4.07                 6.20                 42.86

Feb. 26, 1970       79.27              1.92                 7.59                 44.36
                    79.55     79.26    3.16       2.71      6.34       7.07      48.26       46.31
                    78.97              3.04                 7.28

April 2, 1970       77.35              3.60                 6.41                  41.29
                    77.11     76.92    3.98       3.81      8.30       7.35      39.90       41.49
                    76.29              3.86                                      43.27

April  15, 1970      78.17              2.95                 6.76                 47.66
                    77.71     77.98    2.91       2.84      6.86       6.51      45.04       46.49
                    78.06              2.66                 5.92                 46.76

April 21, 1970      78.92              2.83                 7.28                 43.65
                    78.15     78.27    2.41       2.62      6.58       6.71      42.41        44.41

-------
                                                  TABLE  I (continued)
00
                                                                   % Nitrogen (b)
^Carbon (b)

Date
April 28, 1970 '

May 13, 1970

June 9, 1970

June 22, 1970

July 21, 1970

August 2, 1970

November 13, I970(a)

January II, 1971 (a)

March II, 1971 (a)

fff V ^— ' 1 <-» • i
Each
Samp 1 e
76.97
78.37
50.71


78.85
79.79
79.64
76.40
76.67
75.99
77.71
78.91
78.54
75.37
74.24
79.32
78.58
78.54
78.90
79.25
78.57
79.69
80.90
79.48
79.82
Each
Avq. Sample Avg.
2.60
68.68 2.91 2.85
3.04
2.92
2.83 2.79
2.63
2-45
79.43 2.31 2.37
2.36
2.42
76.35 2.69 2.50
2.40
2.33
78.39 1.68 2.40
3.20
2.72
76.31 2.79 2.46
1.88
2.46
78.67 2.32 2.40
2.43
2.20
79.17 2.57 2.42
2.49
2.23
80.07 2.25 2.24
2.23
Each
Samp 1 e Avg.
6.93
7.24 7.17
7.35
5.63
5.47 5.64
5.81
6.97
7.26 7.13
7.15
6.45
6.62 6.57
6.64
6.89
6.93 6.82
6.66
6.84
6.57 6.73
6.78
7.85 7.81
7.76

6.72
7.53 7.15
7.19
7.51
7.40 7.44
7.41
Each
Samp le
46.42
43.39

42.52(c)
62.89(c)
8l.56(c)
50.51
45.34
48.56
69.36
45.76
63.68
36.82
38.62
44. 18
39.00
37.35
43.88
43.88
43.72
39.49
46.20
48.94
47.59
46.01
45.81
45.06

Avg.
44.90

62.32(c)

48. 14

59.60

39.87

40.08

42.36

47.58

45.63


-------
                                             TABLE  I (continued)

                    % Volatile Solids  % Phosphorus  (b)      % Nitrogen  (b)      	£  Carbon  (b)

Date


May 20, 1971 (a)




August 3,  1971  (a)  81.75
Each
Sample
81.17
80.93
81.27
81.75
82.09
81 .84

Avg.

81.12


81.89

Each
Sample Avg.
2.21
2.20 2.22
2.25
2.31
2.33 2.29
2.24
Each
Sample
7.51
7.62
6.37
7.74
8.23


Avg.

7.17

7.98


Each
Samp le
45.71
44.70
46.09
43.75
43.95
43.50

Avg .

45.50


43.73

(a)  The average percent calcium In the last 5 samples were respectively:  11/13 - 3.2*:   I/I I  - 2 68*-
     3/11  - 3.5IJ6;   5/20 - \ .80%;  and 8/3 - 2.82%.                                                '

(b)  On a  total  dry weight basis.


(c)  Sample contaminated with gasket pieces from blender.

-------
                                 TABLE 2

                 EFFECT OF REFRIGERATED STORAGE OF WAUKESHA
                 TRICKLING FILTER SLIME ON PHOSPHORUS LEVEL

                                                       %  Phosphorus*
Sample                                               Each
Identification     Condition                         Sample        Avg.

    A3             initial sample untreated           3.40
    A4             initial sample untreated           3.31         3.40
    C6             initial sample untreated           3.50

    C7             initial sample washed              3.51
    C9             initial sample washed              3.45         3.46
    D3             initial sample washed              3.42

    DIG            sample washed after 2 hours         3.24
    E4             sample washed after 2 hours         3.33         3.33
    E6             sample washed after 2 hours         3.43

    E8             sample washed in 24 hours           3.71
    E10            sample washed in 24 hours           3.36         3.47
    Ell            sample washed in 24 hours           3.34

    E13            sample washed after 48 hours       2.97         ,  ln
    E14            sample washed after 48 hours       3.23

    A10            sample washed after 14 days         3.42
    All            sample washed after 14 days         3.27         3.22
    Bl             sample washed after 14 days         2.97

    B5             sample washed after 15 days         3.27
    B6             sample washed after 15 days         3.38         3.35
    B7             sample washed after 15 days         3.39


*0n total dry weight basis.
                               60

-------
                                 TABLE 3
               EFFECT OF ROOM TEMPERATURE STORAGE OF ACTUAL
                TRICKLING FILTER SLIMES ON PHOSPHORUS LEVEL
Sample
Identification

Waukesha -
Sample 1

  C3
  C4
  C5

  C6
  C8

  C9
  CIO
  Cll

  C12
  C13
  E14
  B9
  BIO

  J12
  J13
  H14

  LI
  L2
  L3

Waukesha
Sample 2

  LI
  L2
  L3
  L4
  L5
  L6

  E13
  E14
  E16

  B4
  B6
  B7

  J6
  J7
  J8
Condition
initial sample untreated
initial sample untreated
initial sample untreated
initial sample washed
initial sample washed
sample washed after 3 hrs.
sample washed after 3 hrs.
sample washed after 3 hrs.
sample washed after 24 hrs.
sample washed after 24 hrs.
sample washed after 24 hrs.
sample washed after 72 hrs.
sample washed after 72 hrs.
sample washed after 72 hrs.
sample washed after 8 days
sample washed after 8 days
sample washed after 8 days
sample washed after 15 days
sample washed after 15 days
sample washed after 15 days
initial sample untreated
initial sample untreated
initial sample untreated
initial sample washed
initial sample washed
initial sample washed
sample washed after 24 hrs.
sample washed after 24 hrs.
sample washed after 24 hrs.
sample washed after 7 days
sample washed after 7 days
sample washed after 7 days
sample washed after 14 days
sample washed after 14 days
sample washed after 14 days
% Phosphorus*
Each
Sample  Avg.
3,
3,
  08
  02
3.21
 ,12
 ,29
3.14
3.15
2.88
3.72
3.66
3.77

3.57
3.43
3.33
2.75
2.57
2.65

2.81
2.74
2.85

3.11
3.12
3.03

2.98
2.83
2.95

3.29
2.96
3.07
3.10



3.20


3.06
        3.72
        3.44
        2.66
        2.80
        3.09
        2.92
        3.11
                 I Volatile
                   Solids
                Each
                Sample  Avg.
                74.85
                76.34
                76.14
                75.94
                74.95
                75.49

                73.71
                72.28
                74.18

                69.42
                69.77
                69.68
                75.78
                75.46
                73.39
                69.62
                               61

-------
                            TABLE 3 (continued)
Sample
Identification   Condition
Waukesha -
Sample 3

  H7
  H8
  H9
  L4
  L5
  L6
  All
  C13
  L15

Menomonee Falls
Sample 1	
  HI
  H2
  H3

  H4
  H5
  H6

  E9
  E10
  Ell

  MO
  All
  C13

Menomonee Falls
Sample 2	

  H6
  H4
  H5
  J10
  J12
  J13
  C7
  C8
  C9
initial sample untreated
initial sample untreated
Initial sample untreated
sample washed after 24 hrs.
sample washed after 24 hrs.
sample washed after 24 hrs.
sample washed after 7 days
sample washed after 7 days
sample washed after 7 days
initial sample untreated
initial sample untreated
initial sample untreated
initial sample washed
initial sample washed
initial sample washed
sample washed after 24 hrs,
sample washed after 24 hrs,
sample washed after 24 hrs,
sample washed after 7 days
sample washed after 7 days
sample washed after 7 days
initial sample untreated
initial sample untreated
initial sample untreated
sample washed after 24 hrs.
sample washed after 24 hrs,
sample washed after 24 hrs.
sample washed after 7 days
sample washed after 7 days
sample washed after 7 days
                               % Phosphorus*
                               Each
                               Sample  Avg.
2,
2.
2,

2,
2,
2,

3.
2.
2.
2.
2,

3.
2.
3.

3.
3.
2.
2,
2.

2,
2,
2.
 05
 62
 74

 57
 60
 65

 10
 81
2.91
3.03
2.86
3.02
,94
,94
,92

,44
.88
.38
,62
.47
3.69
,80
,88
,84

,76
,84
,39
2.47
2.61
2.94
       2.97
2.93
3.23
3.59
3.40
3.19
3.24
2.84



2.66



3.28
                 I Volatile
                   Solids
                Each
                Sample  Avg.
75.21
74.53
75.05

73.21
73.29
72.74

70.64
71.14
70.97
        79.58
        79.52
        79.61
79.22
79.16
79.28

77.27
79.67
76.23
79.81
79.75
75.18
78.07
78.81
78.81

75.15
74.57
58.33
74.93
73.08
70.92
        79.57
79.22
77.72
78.25
78.56
                      69.35
                              62

-------
                             TABLE 3  (continued)
 Sample
 Identification
 Hales Corners -
 Sample 1	
   .01
   D2
   D3

   D4
   D5
   D6

   E2
   E3
   E8

   C3
   C8
   C9
   J2
   J4
   J5

Hales Corners  -
Sample 2	

  HI
  H2
  H3

  Jl
  J3
  J9

  Cl
  C2
  C3
                                                   Phosphorus*
 Condition
 initial sample untreated
 initial sample untreated
 initial sample untreated
 initial sample washed
 initial sample washed
 initial sample washed
 sample washed after 24 hrs.
 sample washed after 24 hrs.
 sample washed after 24 hrs.
 sample washed after 7 days
 sample washed after 7 days
 sample washed after 7 days
 sample washed after 14 days
 sample washed after 14 days
 sample washed after 14 days
initial sample untreated
initial sample untreated
initial sample untreated
sample washed after 24 hrs.
sample washed after 24 hrs.
sample washed after 24 hrs.
sample washed after 7 days
sample washed after 7 days
sample washed after 7 days
Each
Sample
2.00
2.01
2.38
2.48
2.53
2.50
2.64
2.65
2.30
2.73
3.26
2.57
3.19
2.98
2.64
2.36
2.54
2.29
2.26
2.26
2.19
2.43
2.45
Avg.
2.13
2.50
2.53
2.85
2.94
2.40
2.24
2.38
  % Volatile
    Solids
 Each
 Sample  Avg.
 79.01
 82.40
 74.52
                                                2.27
 80.89
 81.13
 71.85

 81.85
 80.29
 80.76

 74.29
 73.68
 73.96
81.72
80.17
81.85

80.87
80.84
81.01

77.56
77.93
78.66
 78.64
 77.96
80.97
73.98
81.25
80.91
78.05
*0n total dry weight basis
                             63

-------
                                 TABLE 4
                EFFECT OF 35°C STORAGE OF ACTUAL TRICKLING
                     FILTER SLIMES ON PHOSPHORUS LEVEL
Sample
Identification   Condition
                                                  Phosphorus*
% Volatile
  Solids
Waukesha -
C4
CS
C6
HI
H2
H3
H4
H5
H6
H7
H8
H9
H10
Hll
HI 2
L10
Lll
L12
L13
L14
L15
Waukesha -
Jl
J2
J3
E10
Ell
El 2
Waukesha -
L4
L5
L6
L16
L18
L20
Sample 1
initial sample, untreated
initial sample untreated
initial sample untreated
sample washed after 24 hrs.
sample washed after 24 hrs.
sample washed after 24 hrs.
sample washed after 48 hrs.
sample washed after 48 hrs.
sample washed after 48 hrs.
sample washed after 72 hrs.
sample washed after 72 hrs.
sample washed after 72 hrs.
sample washed after 6 days
sample washed after 6 days
sample washed after 6 days
sample washed after 7 days
sample washed after 7 days
sample washed after 7 days
sample washed after 14 days
sample washed after 14 days
sample washed after 14 days
Sample 2
initial sample untreated
initial sample untreated
initial sample untreated
sample washed after 8 days
sample washed after 8 days
sample washed after 8 days
Sample 3
initial sample untreated
initial sample untreated
initial sample untreated
sample washed after 7 days
sample washed after 7 days
sample washed after 7 days
Each
Sample
2.65
2.80
3.23
2.60
2.66
2.69
3.12
3.05
2.44
2.69
2.16
2.95
2.90
2.95
3.03
3.17
3.19
2.93
3.63
2.74
3.15
2.60
2.54
2.61
3.17
3.18
3.38
2.76
2.92
2.64
3.16
3.30

Avg.

2.89


2.65


2.87


2.60


2.96


3.10


3.17


2.58


3.26


2.87


3.26
                                                3.32
Each
Sample
73.74
74.21
72.53
71.73
7.1.00
70.54
70.71
69.69
70.35
68.33
68.67
68.09
66.33
65.84
66.48
66.22
66.36
66.04
63.26
62.10
62.03
71.79
73.93
68.38
67.59
67.68
72.94
72.01
72.97
68.17
70.05.
71.12

Avg.

73.49


71.09


70.25


68.36


66.22


66.21


62.46


72.86

67.88


72.64


69.78

                              64

-------
                            TABLE 4 (continued)
 Sample
 Identification   Condition
 Saukville
  J4             initial sanple untreated
  J5             initial sanple untreated
  J6             initial sanple untreated
  J16            sanple washed after 7 days
  J17            sanple washed after 7 days
  J18            sanple washed after 7 days
 Cedarburg - Sanple 1
  El             initial sanple untreated
  E2             initial sample untreated
  E3             initial sample untreated
  E4             sample washed after 8 days
  E5             sanple washed after 8 days
  E6             sample washed after 8 days
  E7             sanple washed after 19 days
  E8             sample washed after 19 days
  E9             sample washed after 19 days
 Cedarburg - Sample 2

                 initial sample untreated
                 initial sample untreated
                 initial sanple untreated
                 sample washed after 7 days
                 sanple washed after 7 days
                 sample washed after 7 days
  D4
  D5
  D6
  D16
  D17
  D18
Germantown

  H4
  H5
  H6
  H16
  H17
  H18

Menomonee Falls
  HI
  H2
  H3
  D10
  Dll
  D12
                 initial sanple untreated
                 initial sanple untreated
                 initial sample untreated
                 sample washed after 7 days
                 sanple washed after 7 days
                 sample washed after 7 days


                 initial sanple untreated
                 initial sample untreated
                 initial sample untreated
                 sample washed after 8 days
                 sample washed after 8 days
                 sanple washed after 8 days
                                                I Phosphorus*
                                                iiach
                                                Sample  Avg.
                                                2,
                                                2,
                                                2,
  ,34
  ,21
  ,40
2.01
2.32
2.11
                                                2.62
                                                3,
                                                2,
                                                2,
                                                2,
                                                2.
                                                3,
                                                3,
1,
2,
1,
2,
1,
2,
2,
2,
2.
  56
  92
  98
  84
  85
  72
  78
                                                3.38
99
04
95
38
96
2.28
28
40
10
71
2.58
3.08
2.88
2.79
2.85
3.34
3.63
3.28
      2.32


      2.15
      3.03


      2.89


      3.63
1.99


2.21
2.26


2.79
      2.84


      3.42
                                                                   Volatile
                                                                   Solids
Each
Sample
80.42
82.32
82.37
79.66
80.22
78.87
73.24
74.84
75.85
69.38
71.67
68.57
64.74
64.89
64.43
81.41
82.44
82.30
79.01
78.28
78.29
69.10
75.31
68.05
68.80
69.66
68.53
78.69
78.52
78.37
74.94
75.56
75.30

Avg.

81.70


79.58


74.64


69.87


64.69


82.05


78.53


70.82


69.00


78.53


75.27

*0n total dry weight basis
                               65

-------
                                TABLE 5
8 days



9 days

11 days

12 days
                 EFFECT OF ANAEROBIC STORAGE AT 35 C OF
                WAUKESHA PLANT SLIME ON PHOSPHORUS LEVELS
                               (RUN NO. 1)

Time
Initially

1 day

2 days
4 days



5 days




6 days




Supernatant
Phosphorus (mg/1)
Each Sample ^Avg.


13.98* -,
15. 601 14.79X
86. 501 86.50*
190.0* ,
170. Of 167.0
159.07
149. 0
167. 0*
188. Of -I
179. Of 152. 6-1
175. 0:f
154.0
205. O1
200. Of -,
191. Of 191. 51
213.07
174. Of
166. Of
Slime
Phosphorus* (1)
Each
Sample Avg.
1.92i T
3.16f 2.711
3.041












2.77* 1
2.88f 2.841
2.881



% Volatile
Solids
Each
Sample Avg.
79.27* ,
79.55f 79.26-1
78. 971












76.96* ,
77.80f 77. S31
77. 821



183.0,
174.0,
185.0,
184.0,
176.0,
180.0

176.0^
220.0'
213.0'
213.0'

174.0
150.0

180.0'
171.0'

200.0'
200.0'
                           180.3'
2
      205.5'
      162.0'
      175.5'
                            200.0
                              66

-------
 18  days


 21  days


 22  days



 23  days

 25  days

 26  days
                              TABLE 5 Ccontinued)
Time
13 days
15 days
16 days
Supernatant
Phosphorus One/I")
Each Sample
141.0*
155. 0*
192. O2
188. 0^
164. O2
Avg.
148. O2
190. O2
i
Slime % Volatile
Phosphorus* C%) Solids
iiach Each
Sanple Avg. Sample Ava.

164.o:
151.0'
180.Oi
169.0^
200.0^
200.0^
151.0'
                            159.7'
174.5'
200.0'
151.0'
170.O2     170.O2
                        4.292
                        3.71,
                        3.93^
                       3.98'
71.34'
71.70'
71.52'
71.52'
125.0'
125.0'
*Qn total dry weight basis
(1) First flask
(2) Second flask
                                67

-------
                                 TABLE  6
Time

initially
1 day
4 days
5 days



11 days



12 days



14 days
22 days
                    EFFECT OF ANAEROBIC STORAGE AT 35C
               OF WAUKESHA PLANT SLIME ON PHOSPHORUS  LEVELS
                                (RUN NO. 2)
                 Supernatant
               Phosphorus
Each
Sample
7.91*
8.80^
*,
10.06?
0.68;:
0.83
91.76*
83.51,
89.15;:
83.77|:
209. 76 J
151.72,
160.56^
159. 20,
148. 68Z
203.00?
212.56^
189. 48Z
191.56^
181.36,
188.96°
175.00^
188.48,
183.36^
180.88T
181. 36 J
175. 484
127.8oi
121. 10 J
134. 601
*%
113.10?
127.80^
140. 10^

Avg.










180. 741
?
156. 15Z

ry
201. 68Z

187. 293

182. 283

179. 244

127. 834


127. OO5

    Slime
Phosphorus*
Each
Sample      Avg.

3.607
3.987           ,
3.861       3.811
3,
2.
2.
  48
  95
3.02^
3.29,
2.93^
3.58:
3.54:
3.18*
3.39
4.00^
3.96
            3.14J
            3.08'
                                                   3.43^
                                                   3.78"
                            Volatile
                            Solids
                        Each
                        Sample

                        77.35J
                        77.li:
                        76.291
                        69.28?
                        68.97,
                        69.07^
                        64.42^

                        65!03^
AVE
                                                                           76.92J
69. 15J
70.19J
69. 541
72.74?
72. 80,
72.36^
69. 631

72. 632

69. ir
65.024
                                68

-------
                            TABLE 6 (continued)

                 Supernatant               Slime                  % Volatile
               Phosphorus  (mg/1)       Phosphorus*  (%)              Solids
               Each                    Each                     Each
Time           Sample      Avg.        Sample      Avg.         Sample     Avg.

25 days        121.10<         ,       3.97JJ            ,-        64.10$
               125.80^   124.90b       3.79^        3.S25        63.53^     63.13
               127.80s                 3.7CT                    61.75s
*0n total dry weight basis
(1) (2) (3) (4) and (5) refer to flask number
                               69

-------
                                 TABLE 7
Time

initially



1 day



3 days

4 days

13 days
                    EFFECT OF ANAEROBIC STORAGE AT 35C
                 OF LABOi
-------
                                    TABLE 8




EFFECT OF ANAEROBIC STORAGE AT 35°C OF WAUKESHA PLANT SLIME ON PHOSPHORUS LEVELS
Supernatant
Phosphorus (mg )
Time
$
initial




1 day


2 days





3 days





Flask

1
2
3
4
5
1
2

2


3


3


4


Each
Samp 1 e Avg .

0
1
1
1
1
18
20

44
43
43
47
48
48
56
55
62
58
59
61

.796
.454
.154
.633
. 190
.44 18.44
.03 20.03

.32
.10 43.50
.09
.07
.26 47.84
.11
.97
.30 58.09
.01
.60
.09 59.85
.87
%
Phosphorus* % Sol i
Each Each
Sample Avg. Sample

2.
2.
2.


2.
2.
2.
2.
2.
2.



2.
2.
2.




42 7.07
69 2.50 7.06
40 7.14


17
25 2.18
18
15
07-, 2. ! 1
6CT



13
23 2.13
02



% Volatile
ds Solids %
Each
Avg. Sample

76.
7.09 76.
75.


75.
74.
75.
74.
44.
73.



71.
72.
72.




40
67
99


73
87
58
Jt
5I#
60



79
54
00



Carbon
Each
Avg . Samp 1 e Avg .

69.
76.35 45.
63.


43.
75.39 44.

44.
73.94 39.
41.



39.
72.11 36.
42.




36
76 59.66
68


76 44.04
32

89
83 41.95
14



92
75 39.67
39



% Nitrogen
Each
Samp 1 e

6.45
6.62
6.64


6.04
6. 15
5.98
5.70
5.67
5.75



5.51
5.52




Avg.


6.57




6.06


5.71




5,52






-------
                                                     TABLE 8 (continued)
     Tfme    Flask

     4 days    4
     7 days
Supernatant
Phosphorus (mg ) %
Each
Samp 1 e
61.
62.
64.
61.
63.
61.
67.
67.
71.
70
38
81
13
50
82
08
08
97

Avg.

62.96

62.15



68.71

Phosphorus* % Solids
Each Each
Sample Avg. Sample Avg.
2.
2.
2.



2.
2.

10
17 2.15
17



04 2.04
03

% Volati le
Solids %
Each
Sample
72.
72.
72.



72.
71.
70.
19
50
90



05
37
04
Carbon
Each
Avg. Sample Avg,
38.
72.53 32.




33.
71.15 36.

00 35.18
36




26 34.93
60

% Nitrogen
Each
Sample
5.
5.
4.



5.
5.

42
42
92



08
35

Avg.

5.25




5.22


ro
     *0n a percent dry weight basis
     #Problem with analysis procedures - not used in average calculation
     SAverage present calcium in the original sample was 2.78%

-------
                                             TABLE 9

         EFFECT OF ANAEROBIC STORAGE AT 35°C OF WAUKESHA PLANT SLIME ON PHOSPHORUS LEVELS



Time
initial



I day

2 days



Flask
1
2
3
4
1

1
Supernatant
Phosphorus (mg)
Each
Sample Avg.
1.159
1.144
0.7128
2.214
19.256 J9.52
19.787
36.35

% Phosphorus*
Each
Sample Avg.
2.33
1.68 2.40
3.20





% Sol ids
Each
Sample Avg.
6.64
6.49 6.56
6.56




% Volatile
Sol ids
Each
Sample Avg.
77.71
78.91 78.39
78.54





% Carbon
Each
Sample Avg.
36.82
38.62 39.87
44.18





% Ni
Each
Samp
6.89
6.93
6.66





trogen

le Avg.

6.82





                 34.48  34.51
                 32.70

3 days    I       44.68
                 44.68  44.06
                 42.83

6 days    I       59.98
                 59.98  60.66
                 62.01

7 days    I       60.14              1.73                          76.12          45.04           5.89
   '              59.65  59.76       1.81     L82                  75.98   76.13  46.84   44.65   5.79    5.71
                 59.49              1.94                          76.28          42.08           5.45
          2       62.57
                 59.89  62.04
                 63.81

-------
TABLE 9 (continued)
Supernatant % Volatile
Phosphorus (mg) % Phosphorus* % Solids Solids % Carbon % Nitrogen

Time
8 days


9 days


12 days


13 days





14 days


15 days
—

30 days






Flask
2


2


2


2


3


3


3


3


4


Each
Sample
66.27
65.19
65.51
57.05
60.89
58.25
57.83
57. 37
58.91
58.94
61.55
49.75
62.09
60.71
55.85
53.59
53.01
58.88
59.24
60.71
63.17
51.58
50.06
52.01
48.92
49.60
53.64
Each Each Each Each Each
Avg. Sample Avg. Sample Avg. Sample Avg. Sample Avg. Sample Avg.

65.66


58.72


57.86

1.64 73.53 44.96 5.98 6.27
56.75 2.00 2.22 74.19 74.17 46.54 45.80 6.56
3.02 74.80 45.90

59.53


55.13


61.04

2.39 2.36 70.72 40.58 4.50
51.22 2.33 70.98 70.80 33.90 40.05 5.78 5.32
70.69 45.68 5.69

50.72


-------
Time    Flask



6 weeks   4
*0n a dry weight basis
                                                IABLE 9 (continued)
Supernatant
Phosphorus (mg)
Each
Samp 1 e
50.96
48.65
53.32
Avg.
50.98
% Phosphorus*
Each
Samp 1 e Avg .
2.31
2.76 2.61
2.75
% Volatile
% Solids Solids % Carbon
Each Each
Samp 1 e Avg . Samp 1 e
69.80
69.34
69.84
Each
Avg. Sample
42.71
69.66 43.05
41.29
Avg.
42.35
% Nitrogen
Each
Samp 1 e
6.09
5.65
5.78
Avg.
5.84

-------
                                             TABLE 10
         EFFECT OF ANAEROBIC STORAGE AT 35 C OF LABORATORY SLIME ON PHOSPHORUS LEVELS
Time



initial
I  day
2 days
3 days
4 days
7 days
Supernatant
Phosphorus (mg)
Each
Disc Sample Avg.
1
II
1 II
IV
Comp. 	 	
1 1.048
11.087 11.437
12.176
14.109
14.840 14.596
14.840
15.179
15.676 15.493
15.626
16.429
15.725 15.959
15.725
12.613 14.068
15.523
% Vo 1 at i 1 e
% Phosphorus* % Solids Solids % Carbon % Nitrogen
Each Each Each Each Each
Sample Avg. Sample Avg. Sample Avg. Sample Avg. Sample Avg.
.50 1.50
.42 1.40
.38
.53 1.50
.47
.54 1.54
.38 1.74 49.54 9.66
.17 1.30 1.78 1.74 51.22 50.42 9.83 9.70
.34 1.71 50.51 9.63






-------
                                                TABLE
Supernatant % Volatile
Phosphorus (mg) % Phosphorus* % Sol ids Sol ids % Carbon % Nitrogen

Time Disc
8 days*


9 days


10 days


13 days

14 days


15 days


16 days


31 days


6 wk-l day


Each
Samp 1 e
19.154
17.542
17.947
17.363
17.789
17.466
17.363
18.002
17.412
17.734
16.839
16.533
16.429
17.204
19.208
19.099
18.324
17.521
18.056
19.661
20.240
18.002
17.576
14.889
15.921
15.424
Each Each Each Each Each
Avg. Sample Avg. Sample Avg. Sample Avg. Sample Avg. Sample Avg.

18.214


17.539


17.592

17.286


16.722


18.877


18.412


18.606

0.88 97.95
I5.4M 0.80 0.83 95.79 97.44
0.82 98.57
*Percent dry weight basis
#RemaInder with composite sample

-------
                             TABLE I I

                        TOTAL HARDNESS OF
                     TRICKLING FILTER SLIMES


Cl.    c   ,                    o  -J-.L-                  % Hardness#
Slime Sample                  Condition                  „ nn
         1                     —————               as L/dUU —
Waukesha                      initial                    8.3
(Sample l-Table4)            24 hours                  11.4
                              7 days                    13.I
                              14 days                    9.8

Waukesha                      initial                   12.0
(Sample 2-Table 4)            8 days                    12.3

Waukesha                      initial                    9.6*
(Table 5)                      6 days                    10.0*
                              22 days                   14.57*

Cedarburg                     initial                   11.97
(Sample l-Table 4)            8 days                    12.0

Menomonee Falls               initial                   16.1
(Table 4)                      8 days                    14.2

Laboratory Slim© from         initial, Disc  III         1.02*
 Settling Cone                initial, Disc IV           1.17*
*Slime digested with HCi.  All the other samples digested with perch-
  loric-nitric  acid mixture according to Standard Methods.

#After digesting a known weight of oven dried slime, the sample was
 so I Lib I i zed to a convenient volume, usually 50 ml.  The hardness con-
 centration in this volume was determined and expressed as mg/l CaCO,.
 This concentration was then expressed as a weight and related to the
 original weight of the dried slime sample, hence the unit,
 ^Hardness as CaCO,.
                             78

-------
       Run
\D
       I-A-S-I
       I-B-S-I
                                 TABLE 12

            LOG OF ALL TEST RUNS CONDUCTED WITH DISC APPARATUS

                                             Phosphorus
        Speed           Carbon     Nitrogen   NaJHPO     Calcium
 Date   RPM    Water  COD (mg/l)   N (mg/l)   P.fmg/T)  (mg/l
                   1-21-70  I
                   1-30-70  I
                   2-11-70  I
2-26-70  I
3-31-70  I
               Soft
               Soft
               Soft
Soft
Soft
Glucose NH Cl
300 12
Glucose NH.CI
300 12
Glucose NH Cl
300 12
Glucose NH Cl
300 12
Glucose NH.CI
300 12
Glucose NH Cl
300 12
.24,
12,
.24,
12,
.24,
12,
.24,
12,
.24,
12,
.24,
12,
2.4,
24
2.4,
24
2.4,
24
2.4,
24
2.4,
24
2.4,
24
                                                         *        $
                                                    Light  Remarks
No    Same yield; l-yellow;
      II, Ill-white; IV-pink;
      I-thickest growth

No    I-pi nk; I I-It. p ink;
      III, IV-cream; enough
      growth for duplicates
      on VS,  P,C,N;  l-thickest
      growth

No    I, II,  III, IV-creamy
      white;  I Il-thickest
      growth; IV-most
      cohesive

No    I-white;  I  I-creamy
      white;  III, IV-creamy
      white with little green;
      IV-most cohesive; 1-40 ml;
      I  1-45 ml;   !I 1-80 ml;
      IV-50 mKSIime yields)

No    I, II,  I I I-white;
      IV-dark cream; I I-most
      cohesive

No    All white;  I-most
      cohesive;   I I-very
      fragi le

-------
                                                  TABLE 12 (continued)
     Run

     I-A-S-IOO
 Date
Speed
RFM
Water
                                     #
4-24-70  100   Soft
Carbon
COD (mg/l)
G 1 ucose
300
Nitrogen
N (mg/ 1 )
NH.CI
A
Phosphorus
Na7HPO
P fmg/T)
.24, 2.4,
12, 24
Ca 1 c i urn
(mg/l )Ca

      I-B-S-IOO
 5-8-70  100   Soft
                Glucose
                   300
o>
o
      l-A-S-50
5-22-70  50
       Soft
         GIucose
            300
      [-B-S-50
 6-5-70  50
       Soft
         GIucose
            500
                     NHC,
NH Cl
 12
                                                            No
           .24,  2.4,
           12, 24
                     No
.24,2.4,
12,  24
No
           .24,2.4,
           12, 24
No
Rema rks

All units cohesive; very
thin growth; I I-very pale
green algal growth at
edge; Ill-grey-brown;
IV-black specks; II &
IV-granular

I-cream; II, IV-yellowish
with some algae; Ill-cream
with some algae; IV-most
cohesive;  ll-thickest
growth

I-white with faint pink;
I-white with taint pink;
I I-yeI low with traces of
a Igae;  I I I-ye I Iow-green;
IV-yellow;  all very
fragile;  II, Ill-granular
IV-thickest, but a lot
broken off.
                           l-white with light pink;
                           II,  IV-creamy white to
                           yellow with green around
                           edges;  Ill-pale green
                           with dark green at edges;
                           II,  III- granular;  II,  IV-
                           s lough off; 1-35 ml ;
                           I 1-80 ml; I I 1-25 ml;
                           IV-50 ml

-------
Run
Date
TABLE 12 (continued)
             Phosphorus
                         Ca I c i urn
                        (mq/DOa
ll-A-S-25  6-18-70   25
ll-A-S-50  7-1-70
M-B-S-25  7-20-70   25
Speed
RPM
25





50





100





25





„ Carbon
Water* COD (mg/l)
Soft Glucose,
Nutrient
Broth;
Yeast-
Extract
330
Soft Glucose;
Nutrient
Broth;
Yeast
Extract
330
Soft Glucose;-
Nutrtent
Broth;
Yeast
Extract
330
Soft Glucose;
Nutrient
Broth;
Yeast
Extract
330
Nitrogen
N (mg/l)
Glucose;
Nutrient
Broth;
Yeast
Extract
22-24
Glucose;
Nutrient
Broth;
Yeast
Extract
22-24
Glucose;
Nutrient
Broth
Yeast
Extract
22-24
Glucose^
Nutrient
Broth;
Yeast
Extract
22-24
Na HPO
P fmg/T)
3, 6, 9,
12
+ from
feed


3,6,9,
12
+ feed



3,6,9,
12
+ feed



3,6,9,
12
+ feed



Remarks5
                                                                   No     I-pinkish yellow;  II,  III,
                                                                          IV-yellow with green
                                                                         algae;  I, Ill-feathery;
                                                                          I I-most cohesive;  I,  III,
                                                                          IV-partly sloughed off;
                                                                          1-25 ml; I 1-75 ml;  I I I-
                                                                         25 ml;  IV-60 ml

                                                                   No     I-orange; II,  III,  IV-
                                                                         yellow orange with algae
                                                                         growth;  I-feathery and
                                                                         filamentous;  1-50 ml;
                                                                          11-100 ml;  I I 1-50 ml;
                                                                          IV-75 ml

                                                                   No     I-pink;  II,  I I I,  IV-
                                                                          Iight pink with some
                                                                         algae; very thin growth;
                                                                         smal I yield;  I, IV,.-10 ml;
                                                                          I I,- I I 1-25 ml.
                                                                   No     l-pink;  II,  Ill-pinkish
                                                                         orange with  small amount
                                                                         atgae; tV-yellow with
                                                                          little algae;  1-25 ml;
                                                                          It,  I I 1-50 ml;  IV-30 ml

-------
       Run

       I Il-A-S-25
       Speed
Date   RPM

7-28-70  25
     jt   Carbon
Water  COD (mg/l)

Soft
TABLE 12 (continued)
             Phosphorus
  Nitrogen    Na HPO    Calcium
  N (mg/l)    P fmg/T) (mg/l)Ca
       lll-B-S-25   8-5-70   25   Soft
o>
ro
       III-A-S-IOO  8-21-70  100  Soft
       IV-A-S-25
9-4-70   25   Soft
                       Glucose;
                       Nutrient
                       Broth;
                       Yeast
                       Extract
                         660
Light* Remarks
Glucose
Nutrient
Broth;
Yeast
Extract
660
Gl ucose;
Nutrient
Broth;
Yeast
Extract
660
G ! ucose
Nutrient
Broth;
Yeast
Extract
40-45
G 1 ucose ;
Nutrient
Broth;
Yeast
Extract
40-45
3,6,9,
12
+ feed



3,6,9,
12
+ feed



                     Glucose;
                     Nutrient
                     Broth;
                     Yeast
                     Extract
                       40-45
              3,6,9,
              12
              -I- feed
Mi Ik
320
Milk
14-15
from
mi Ik
only
from
mi 1 k
only
 No    All  creamy yellow;
       feathery;  I-cohes i ve;
       II,  III, IV-slough  off;
       II,  IV-slimy;  I,  II-
       50 ml;  I I 1-40 ml;
       IV-60 ml

 No    AM  creamy yellow-orange;
       II,  III, IV-1i ke peach-
       skin in thin part;  I,
       Ill-ropey  in thick  part;
       1-40 ml; I 1-55-60 ml;
       I I 1-30 ml; IV-50 ml;
       2 different thicknesses
       due to motor stopping
       over-night

 No    1,11,11 I-rust color;
       IV-rose color;  very
       blotchy, large chunks,
       broken off;  thick and
       thin sections on each
       disc;  1-35 ml;  I 1-55 ml;
       I I 1-30 ml; IV-50 ml

 No    I,II,I I I-peach;  IV-cream;
       I,IV-variable thickness;
       I 1, 11 l-uniform thickness;
       J,  IV-stringy;  I-tenacious;
       I I,I Il-firm siime;  IV-
       fragile

-------
                                                   TABLE 12 (continued)
      Run

      V-A-S-25
 Date
Speed
RPM
9-11-70  25
Oo
      V-A-S-IOO   9-25-70  100
      V-B-S-IOO  10-9-70   100

Water*
Soft


Soft


Soft


Hard

Hard


Carbon
COD (mg/l )
Milk
320

Milk
320

Milk
320

Milk
320

Milk
320


Nitrogen
N (fpg/l)
Milk
14-15

Mi Ik
14-15

Milk
14-15

Milk
14-15

Milk
14-15

Phosphorus
Na9HPO
P tmg/T)
3,6,9,
12
+ feed
3,6,9,
12
+ feed
3,6,9,
12
+ feed
3,6,9,
12
+ feed
3,6,9,
12
+ feed

Calcium
(mg/l )Ca
from
mi 1 k
only
from
mi Ik
only
from
mi Ik
only
CaCI7
100
+ mi Ik +
CaCI7
100
+ mi Ik +
Remarks
                                                            No     I-peach;  I I, I I 1,IV-cream;
                                                                   I-uniform thickness;
                                                                   11,111,IV-irregular
                                                                   thickness;  I-cohesive;
                                                                   IV-very small  specks of
                                                                   green;  1-80  ml;  I I,IV-
                                                                   100 ml; I II-7C ml

                                                            No     I,II,IV-cream;  Ill-
                                                                   reddish;  1,111-80 ml;
                                                                   I 1-100 ml;  IV-70 ml

                                                            No     I, I I, I Il-tannish pink
                                                                   slime; very  tenacious;
                                                                   IV-yeI low-green and
                                                                   fragile

                                                            No     All orange;  rough textured;
                                                                   different  from all other
                                                                   runs; 1,11,11 I-tenacious,
                                                                   dry-consistency of squash;
                                                                   III,IV-gritty;  1-55 ml;
                                                                   11-80 ml;  IIl,IV-75 ml

                                                            No     1,11,Ill-variegated, light
                                                                   and dark brown;  IV-same
                                                                   with green;  1,11,111-
                                                                   tenacious;  11,Ill-dry;
                                                                   I 11-gritty at edge; IV-
                                                                   gritty all over; 1-75 ml;
                                                                   11-90 ml;  I I 1-50 ml;
                                                                   IV-85 ml

-------
                                                  TABLE 12 (continued)
                                                              Phosphorus
     Run
Date
     VI-A-H-IOO  11-25-70
CD
     VII-B-S-25  I-II-7I
Speed
RPM
i 100


100

I 25

25

„ Carbon
Water COD (mg/l)
Hard Milk
320


Hard Milk
320

Soft Milk
320

Soft Milk
320

Nitrogen
N (mg/ 1 )
Milk
14-15


Mi Ik
14-15

Milk
14-15

Milk
14-15

Na7HPO
P Tmg/T)
3,6,9,
12
+ feed

3,6,9,
12
+ feed
9
+ feed

9
+ feed

Ca 1 c ! urn
(mg/l)Ca
CaCI7
100
+ mi Ik +
H20
CaCI7
100
+ mi fk
CaCI?
0,25750,
100
+ mi Ik
CaCI2
0,25,50,
100
+ milk
Ligl
No


No

No

No

  *        $
ht* Remarks

    IV-Iight pink;  l-dry;
    all  very tenacious;
    III,!V-come off in
    sheets; 1-70 ml;  II-
    75 ml;  II1-65 ml;  IV-
    55 ml

    1,11l-dark yellow-brown;
    I I-1ight yellow-brown;
    IV-yellow-orange with
    brown blotches; all  very
    tenacious;  1,11,Ill-very
    dry;  1-60 ml;  11-50 ml;
    III,IV-70 ml

    I-cream-darkest of four;
    all  good thick growths;
    I 1,111,1V-a11 with a  lot
    sloughed off and lumpy;
    I-jeI IyIi ke cons i stency;
    11,111,IV-watery
    consistency;
    I,IV-250 ml; 11,111-
    200 ml

    l-beige; ll-orange (darkest);
    I I I-between I and I I;
    IV-yellow;  II and IV-very
    cohesive;   I and Ill-less
    cohesive;   1-225 ml;   II-
    125 ml; I 11-90 ml; IV-
    50 ml

-------
                                                    TABLE  12  (continued)
      Run
Date
      VII-A-S-IOO   1-29-71
      VM-A-S-IO   2-12-71
CD
Ul
      VIII-A-H-IO  2-24-71
Speed
RPM
100
10
10
„ Carbon
Water COD (mg/l)
Soft Milk
320
Soft Ml 1 k
320
Hard Milk
320
Nitrogen
N (mg/l)
Mi Ik
14-15
Milk
14-15
Milk
14-15
Phosphorus
Na9HPO.
P Tmg/T)
9
+ feed
9
+ feed
0,3,6,
12
+ feed
Calcf urn
(mg/DCa.
CaCI7
0,25750
100
+ milk
CaCI7
0,25750,
100
+ milk
Ca(OH)
50
+ milk
     #        J
Liqht  Remarks
                                                                   No    I-Iight ye I low;  II-
                                                                         yellow; Ill-orange-
                                                                         yellow; IV-orange with
                                                                         green; I-smooth texture;
                                                                         11,111-texture not
                                                                         consistent;  IV-pumpktn-
                                                                         like consistency; all
                                                                         very cohesive; 1-20 ml;
                                                                         Il,IV-25 ml;  II1-30 ml

                                                                   No    I-white;  1,111-yeI low-
                                                                         beige; IV-peachy; I-soft
                                                                         and smooth-not tenacious;
                                                                         Il-thick and  thin-tenacious
                                                                         where thin;  not where
                                                                         thick; Ill-lumpy, tenacious;
                                                                         IV-tenacious; comes off
                                                                         i n sheet-ski nIi ke;
                                                                         1-100 ml; I 1,111-120 ml;
                                                                         IV-60 ml

                                                                   No    I-peach;  I 1,111,1V-
                                                                         ye11ow i sh;  I-smoothest-
                                                                         but small bumps;   ll-bumps
                                                                         most prominent;  I 11,1V-
                                                                         pebbly appearance;
                                                                         Il,IV-peels off  I ike
                                                                         skin; a 11 cohesive;
                                                                         1,111-175 ml;  11-90 ml;
                                                                         IV-120 ml

-------
00
                                                   TABLE 12 (continued)

                                                               Phosphorus
Speed ., Carbon Nitrogen
Run -Date RPM Water COD (mg/l ) N(mg/l)
VIM-B-H-IO 3-10-71 10 Hard Milk Milk
320 14-15
IX -A-H-IO 3-26-71 10 Hard Milk Milk
320 1 4- 1 5
IX.-B-H-IO 4-14-71 10 Hard Milk Milk
L 320 14-15
IX.-A-H-50 4-30-71 50 Hard Milk Milk
L 320 14-15
P ?mg/f )
0,3,6,
12
+ feed
0,3,6,
12
+ mi Ik
0,3,6,
12
-*• mi Ik
0,3,6,
12
+ milk
Calcium *
(mg/l) Ca Light* Remarks
Ca(OH)2 No 1 -ye 1 low-brown; ll-cream;
50 Ill-grey-green; 1 -uniform
+ milk thickness; others have
+ H?0 sloughed off patches;
l-most tenacious; IV-
slimy; 1- 100 ml; 11,111-
175 ml; IV-150 ml
Milk Yes 1 -green particles in
+ HJD clear or tannish matrix;
11,111, 1 V-tan on green;
tan washes off; 1 V-tan
more stringy; M-bumpy;
under it is tenacious;
1-50 ml; 11,11 1-70 ml;
IV-60 ml
Milk Yes 1 -grey-white over green
+ HO with reddish brown blotches
in green; ll-green with
very few red-brown blotches;
Ill-grey-white over green
with very little brown;
IV- variegated green with
brown bumps; IV-most
tenacious; 1-120 ml;
II,IV-IOO ml; Ill-ISO ml
Milk Yes All dark green with no
+ H~0 white; 1- least bumpy;
IV-most bumpy; IV-drtest
                                                                                            and most cohesive;
                                                                                            l-l10 ml; I 1-120 ml;
                                                                                            111-120 ml;  IV-IOO ml

-------
      Run
Date
Speed
RF"M    Water
CO
-4
      IXL-A-H-IOO   5-19-71   100  Hard
  Carbon
COD (mg/l)

 Mi Ik
   320
                                                  TABLE 12 (continued)

                                                             Phosphorus
Nitrogen
N (mg/l)
Na>IPO.
P fmg/T)
                                   Mi Ik
                                     14-15
                                       0,3,6,
                                       12
                                       + mi Ik
      XL-A-H-IO
6-4-71    10   Hard
      XL-A-H-IOO    6-23-71   100  Hard
                        Mi Ik
                          320
                            Milk
                              14-15
                        0,3,6,
                        12
                        + mi Ik
 Calcium
(mg/l)  Ca

 Mi Ik
Liqht* Remarks
                                                                                                  $
Mi Ik
320

Milk
14-15

0,3,6,
12
+ mi Ik
CaCI
100
+ mi Ik
                      CaCI7
                      100
                      + mi Ik
                      + HO
                      Yes   I-light tan-green over
                            dark green;  11,1 Il-bright
                            green blotches over grey-
                            green;  IV-finely mottled-
                            light and dark green;
                            1-dry;  I I,Ill-more wet;
                            IV-thick and lumpy-
                            consistency of thick milk
                            shake,  very tenacious;
                            1-150 ml;  I 1-120 ml;
                            I I 1-90 ml; IV-60 ml

                      Yes   All  four  light green
                            filamentous growth over
                            inner 2/3 of discs; IV-
                            most cohesive,  then I  -
                            III  - II;  I-smooth; I I-
                            more fluid;  IV-dry-gritty;
                            1-175 ml;  I 1-140 ml,
                            I Il-l10 ml;  IV-90 ml

                      Yes   Under layers-green;
                            tenacious; upper layer
                            Iighter-not very
                            tenacious; I  and I  I -
                            pebbly looking;  1-100 ml;
                            I 1-105 ml; I 11-95 ml;
                            IV-60 ml

-------
                         Speed
     Run          Date   RPM

     VIML-A-H-IO 7-2-71   10
          Carbon
Water*  COD (mg/l)  N (mg/l)

Hard
        TABLE  12  (continued)
                   Phosphorus
          Nitrogen   Na0HPO
Milk
  320
Mi Ik
  14-15
QO
oa
0,3,6,
12
+ milk
                     Calcium
                     P Tmg/T)  (mg/l)Ca   Light* Remarks1
CaO
50+
mi Ik +
H20
Yes   I-brown-green; 11,111,
      IV-green; I-most
      tenacious, 11  and III-
      least tenacious; I  and
      IV-f iIamentous growth
      moves about when washed;
      I I  and I  I I-top layer
      washes off;
      l-like very thick taffy;
      It  and IV-1 ike taffy;
      Ill-soft
    V1IIL-A-H-IOO 7-19-71   100  Hard
          Mi Ik
            320
           Milk
             14-15
     i Soft =  Milwaukee tap Zeolite softened
       Hard =  Milwaukee tap
     * Plant light on continuously over disc apparatus
     $ Slime yield In ml reported where applicable
           0,3,6,    CaO        Yes   I I-most tenacious; not
           12        50+              moved with H^O; I  and III-
           + milk    milk+            move with H.-0 wash; IV-
                     HJD              lightest green; I-ye I low-
                                      green with brown;  ll-dark
                                      ye I low-green with  dark
                                      green bumps; Ill-light
                                      green in middle, dark
                                      green at edges; 1,11,111-
                                      consistency of baby food.

-------
                                                    TABLE 13

                                   ANALYSIS OF DISC SLIMES AT VARYING SPEEDS
                                       AND DIFFERENT GROWING CONDITIONS
00
\o
      Date and
      Identification

      Jan. 14, 1970
       X-A-S-I*
Jan. 21. 1970
X-B-S-lf
      Jan. 30, 1970
       X-C-S-I#
Disc

I
II
III
IV

I
                  I II

                  IV

                  I


                  11


                  III


                  IV
% Volatile
Each
Sample
94.68
91.95
91.78
93.72
94.09
96.76
91.33
90.44
95.41
89.29
90.92
90.10
96.70
93.67
96.00
94.59
94.21
93.41
93.76
92.25
92.27
91.17
89.73
90.62
Solids

Avg.
94.68
91.95
91.78
93.72

95.42

90.88

92.35

90.55

95.46


94.07


92.76


90U5I

% Phosphorus*
Each
Samp 1 e
D.3I
1.80
1.81
1.51
D.22
).22
.82
.84
2.10
2.10
.78
1.85
D.I 1
D.ll
D.ll
.15
.09
.12
.20
.25
.07
.22
.23
.30

Avg.
0.31
1.80
1.81
1.51

0.22

1.83

2.10

1.81

0. II


1.12


1.17


1.25

% Carbon
Each
Sample
51.00
58.91
53.66
51.67
45.72
45.34
28.95
44.47
51.79
47.70
48.11
33.34
23.10
24.97
27.71
41.44
40.82
42.90
39.04
44.62
40.69
46.12
47.51
34.09

Avg.
51.00
58.91
53.66
51.67

45.53

36.71

49.74

40.72

25.26


41.72


41.45


42.84

% Nitrogen
Each
Samp 1 e
4.06
7.39
8.44
5.89
3.60
3.82
9.62
9.81
11.18
11.07
9.98
9.83
4.17
4.15
3.90
7.02
5.79
6.52
8.75
8.55
8.60
8.77
8.44
8.86

Avg.
4.06
7.39
8.44
5.89

3.71

9.71

11.12

9.90

4.07


6.44


8.63


8.69


-------
                                                    TABLE 13 (continued)
VO
o
     Date ana
     Identification

     Feb.  II,  1970
      X-D-S-I#
      Feb.  26,  1970
       I-A-S-I#
Disc
                        I I I
                         II
      March 31,  1970    All
       I-B-S-I
      April 2, 1970**   I
                        I I I
% Volati le
Each
Sample
95.80


92.00


90.34


91.72


73.91


84.53

83.01


87.50


95.63
94.85
96.10


94.71
93.75

Sol ids

Avg.

95.80


92.00


90.34


91.72


73.19


84.53

83.01


87.50


95.53




94.23

% Phosphorus* %
Each
Samp 1 e
0.33.
0.46
0.37
.42;
.64.
.52'
.68.
.15
2.17.
.54
.72'
.40
0.31 .
0.30 '
0.33
.31
.27
.51
0.76,
.04.
.87
.94' .
2.32
0.76
0.58:
0.79"
2.38
i.92
1.87
1.88
1.70

Avg.

0.39


1.53


1 .67"


1.55


0.31


1.29

1.10


2.04


0.71-

2.38
1.92

1.82

Each
Sample
56.29
45.42

53.47
46.27

50.21
46.50
44.59
49.69
48.52

83.21
88.20

64.31

56.74
49.27

54.90


39.60
49.76
43.39


43.65


Carbon

Avg.

50.85


49.87


47.10

49.10



85.70


64.31

53.00


54.90


44.25




43.65

                                                                        %  N i trogen
                                                                     Each
                                                                     Sample      Avg.
                                                                     4.64
                                                                     4.83
                                                                     4.64
                                                                     8.89
                                                                     8.29
                                                                     8.84
                                                                     9.37
                                                                     9.22
                                                                     9.52
                                                                     9.99
                                                                     9.69
                                                                     9.82

                                                                     4.78
                                                                     4.45

                                                                     6.52

                                                                     7.12
                                                                     7. 13

                                                                     8.08
                                                                     4.48
                                                                     4.54
                                                                     4.62
                                                                                             8.89
4.70


8.67


9.37


9.83



4.61


6.52

7.13


8.08



4.55
                                                                                                        8.89

-------
vO
      Date and
      Identification
      April  24, 1970
       I-A-S-IOO
      May 8, 1970
       I-B-S-IOO
      May 22, 1970
       I-A-S-50#
      June;5,  1970
        I-B-S-50#
% Volatile Solids

Disc
IV

1
II

II 1
IV
1
II

1 1 1
IV


1


II


III


IV


1


Each
Samp 1 e
95.83
96.51
97.62
95.07

85.96
93.89
97.39
90.94
92.16
94.59
89.94
90.59

97.94
97.62
98.38
93.62
93.66
93.84
95.06
96.39

95.27
95.31
93.72
96.34
94.50
91.14

Avg.

96.17
97.62
95.07

85.96
93.89
97.39
91.55

94.59
90.27



97.98


93.71

95.73



94.77


93.99

TABLE 13 (continued)
% Phosphorus* % Carbon
Each
Samp 1 e
1.39
1.23
0.78
1.26
1.35
!.57
1.36
0.27
1.53
1.52

1.50
1.49
1.43
0.51
0.26
0.28
1.75
1.60
1.72
1.43
1 . 36
1.46
1.52
1.50
1.64
0.28
0.44
0.25

Avg.

1.31
0.78
1 .30

1.57
1 .36
0.27
1.53



1.47


0.35


1.69


1.42


1.55


0.32

Each
Samp 1 e
43.99







45.07
47.85

56.17
51 .30

20.32
39.00
27.60
33.1 1
32.14
53.74
39.49


29.62
33.00
37.80
41.74
51.08


Avg.

43.99






46.46


53.73



28.97


39.66

39.49



33.47


46.41

                                                                                                % Nitrogen
Each
Samp Ie

8.51
8.58
8.83
8.76

8.20
8.18
5.
5,
5.
94
30
39
                                                                                             8.73
                                                                                             7.
                                                                                             7.
  84
  97
                                                                                             9.06
9.47
8. !0
9.51
5.53
4.26
4.40
         Avc
                                                                                                        8.51
8.72


8. 19




5.54


8. 18

9.06



9.03



4.73

-------
                                                  TABLE 13 (continued)
     Date and
     Identification
     June 18, 1970
      ll-A-S-25#
\o
to
     July I, 1970
      I I-A-S-50#

Disc
II


III


IV


1


1 1


II 1


IV


1


1 1


1 II


IV


% Volati
Each
Sample
89.32
89.86
89.61
90.26
89.55
89.29
87.50
88.60
88.22
90.58
92.67
90.58
90. 19
91.80
91.64
92.77
91.43
91.91
89. 18
88.76
90.46
91.40
91.03
90.49
89.25
89.67
90.43
89.19
89.20

87.87
90.22

le Solids % Phosphorus*
Each
Avg. Sample

89.60


89.70


88.11


91.28


91.21


92.04


89.47


90.97


89.78

89.20


89.04


.61
.55
.60
.60
.49
.68
.99
.82
.75
.70
.95
.74
.66
.71
.70
.60
.77
.60
.90
.94

.47
.28
.38
.34
.36
.42
.36
.59
.38
.59
.58
.35
Avg.

1.59


1.59


1.85


1.80


1.69


1.66

1.92



1.37


1.37


1.44


1.50

% Carbon
Each
Sample
40.58
59.89
47.51
49.72
47.44
58.01
43.28
48.34
34.65
31.54
51.79
59.59
48.98

45.45
57. 15
56.29
47.96
33.68
51.41
56.62
43.43
42.56
47.44
41.21
46.16

35.29
37.61
43.43
46.84
39.86
48.63
Avg.

49.32


51.76


42.09


47.64


47.22


53.80


47.25


44.48

43.69



38.78


45.11

% Nitrogen
Each
Sample
9.33
9.30
9.05
9.36
9.03

8.96
8.82
8.52
1 1.50
1 1.32
1 1.44
10.69
10.24
10.59
10. 19
10.33
10.01
10.77
11.05
10.25
10.00
10.03
9.91
9.53
9.74
10.02
10.01
10.32
10.22
9.82
10.44
9.66
^g.

9.23

9.20



8.77


1 1.42


10.67


10. 18


10.69


9.98


9.76


10. 18


9.97


-------
     Date and
     Identification

     July 13, 1970
      M-A-S-IOOiJf
vO
     July 20,  1972
      ll-B-S-25
     July 28,  1970
      lll-A-S-25
                       III
I

I I

III

IV
                        &
                       IV
TABLE 13
(continued)
% Volatile Solids -% Phosphorus*
Each Each
Sample Avg. Sample
91.43 92.12
92.82

93.30
93.13 93.31
93.50
92.50
92.49 92.35
92.07
89.35 90.60
91.84

97.95
95.79 97.44+
98.57




92.93
93.25 93.02
92.88
94.23
94.30 94.12
93.82
93.35
93.32 93.36
93.42
.06
.22
.29
.44
.00
.27
.23
.35
.32
.21
.00
.39
.50

.42
.38
.53
.47
.54
.67
.46
.72
,66
.70
.61
.74
.73
.67

Avg.

1. 19


1.24


1.30


1.20

1.50

1.40

1.50

1.54

1.61


1.65


1.71

% Carbon
Each
Samp I e
49.09
88.88

46.00
41.65
41.68
48.79
44.44
44.48
41.32
43.35

49.54
51.22
50.51




33.41
44.28
42.26
38.21
36.38
35.18
40.28
41.25
36.82

Avg.
68.99



43. 1 1


45.90

42.33



50.42+






39.98


36.59


39.45

                                                                                               % Nitrogen
 Each
 Samp Ie

 9.38
10. 10

 9.62
 9.87
 9. 18
 9.81
 9.87
10.12
 9.58
 8.92


 9.66
 9.83
 9.63
                                                                    I 1.36
                                                                    11.07
                                                                    I 1.74
                                                                     9.41
                                                                     8.62
                                                                     9.15
                                                                    10.74
                                                                    10.32
                                                                    10.29
Avg.

9.74



9.56


9.93

9.25




9.704
           I 1.39


           9.06


           10.45

-------
                                                   TABLE 13 (continued)
                                                      % Phosphorus*
              Carbon
vO
Date and
Identification Disc
August 5, 1970 1
lll-B-S-25#

II


1 II


IV


August 21, 1970 1
1 1 I-A-S-IOO#

II


Ml


IV


Sept. 4, 1970 1



Each !
Samp 1 e Avg . I
93.05
93.16 93.08
93.03
93.19 :
93.22 93.22 ;
93.23 :
93.84
93.25 93.43
93.21
93.50
93.29 93.29
93.09
95.35
96.23 95.75
95.69
94.79
94.77 94.67
94.46
94.56 94.86
95.16

94.29
94.47 94.26
94.01
89.92
88.68 89.61
90.22

Each
>amp 1 e
.38
.48
.37
5.09
2.77
2.83
.41
.48
.51
.53
.60
.49
.75
.61
.67
.17
.58
.60
.71
.80
.83
.63
.80
.43
.6la
.563
5,a
'.64*
Each
Avg. Sample Avg.
55.31
1.41 53.51 55.12
56.55
49.58 54.32
2.90 59.06

53.36
1.47 53.51 53.36
53.21
52.09
1.54 50.49 51.28
51.30
57.52
1.68 50.78 55.24
57.41
52.28 51.53
1.45 50.78

52.09 54.34
1.79 56.59

51.68
1.62 53.18 50.57
46.84
44.40
I.573 52.54 44.01
35.10
L
Each
Sample
10.46
9.76
10.08
10.40
11.04
10.44
10.62
10.31
10.32
10.50
10.46

1 1.23
1 1.37

10.26
11.12
11.40
11.20
11.03

11.25
12.41
11.58
10.28
10. 18
10.78


Avg.

10.10


10.63


10.42

10.48


1 1.30



10.93

11.12



11.75


10.41


                                                     1.58:
                                                     1.60
.61

-------
                                                   TABLE  13  (continued)


                                %  Volatile  Solids      % Phosphorus*
vo
vn
Date and
Identification









Sept. II, 1970
V-A-S-25#










Sept. 25, 1970
V-A-S-IOO#










Each
Disc Sample
II 90.04
89.07
90. 19
III 89.99
90.34
89.33
IV 88.38
88.48
88.76
1 91.79
91.53
92.67
II 91.42
92.08
92.06
III 92.86
92.50
93.00
IV 92.72
93.26
93.06
1 93.57
91.27
91.67
II 94.74
92.74
91.23
IN 92.69
91.72
92.66
IV 92.99
92.23
93.87

Avg.

89.77


89.89


88.54


92.00


91.85


92.79


93.01


92.17


92.90


92.36


93.03

Each
Sample
.53
.65
.61
.67
.95

.63
.64
.55
.44
.46
.35
.45
.50
.50
.66
.66
.56
.61
.40
.61
.41
.36
.41
.26
.29
.20
.54
.50
.52
.43
.37
.38
Each
Avg. Sample
46.05
1.60 50.40

1.81
52.05
59.44
49.01
1.61 47.70
48.86
47.44
1.42 51.34
41.89
46.84
1.48 47.25
44.63
47.74
1.63 67.88
46.39
41.28
1.54 28.80
78.30
47.70
1.39 49,39
48. 19
45.90
1.25 45.26
47.44
50. 18
1.52 44.78
46.69
44.10
1.39 46.20
46.73

Avg.
48.22



51.18


48.52


46.89


46.29


54.00


49.46


48.42


46.20


47.20




% N
Each
Samp le
10.61
10.40
9.73
10.79
10.79
10.23
11.16
1 1.20
11.15
8.78
10. 17
10. 17
9.44
8.77
9.33
9.99
10.25
10.25
1 1.08
10.39
10.60
10.75
11.18
9.82
6.40
9.41
10.27
10. 18
10. 18
10.21
9.29
7.64
itrogen

Avg.

10.25


10.60


11.17


9.71


9. 18


10.17


10.69

10.97
• •

8.54


10.24


9.05


-------
                                          TABLE  13  (continued)
                          % Volatile Solids     % Phosphorus*
                                                     % Carbon
% Nitrogen
Date and
Identification
Disc
October 9, 1970   I
 V-B-S-IOO
                  IV
October 29, 1970
 VI-A-H-25#
                  II
                   II
Each E
Sample Avg . S
94.74
93.61 94.39 .
94.82


93.20
92.79 92.98
92.96 ^ *


93.82
92.61 93.04



93.68
93.63 93.82
94.15




87.05 :
86.89 86.68 :
86.10 :
85.42 :
86.10 85.96 :
86.37 i
83.33 :
83.38 83.08 :
82.52 /
iach Each Each
>ample Avg. Sample Avg. Sample Avg.
>na
.41
.36a I.383
l£
!43b l.40b
.40
.50a
.50a I.523
:l$
!58b l.54b
.52
•56!
.62a l.63a
•6*H
n n
,6r 1.62
n
.61
.69a
.49a l.60a
•6lb
•56b b
.54° 1.54°
h
.53°
?.47 43.99 8.65
2.44 2.42 49.12 46.25 8.01 8.34
2.36 45.64 8.35
2.50 39.60 7.78
2.46 2.50 40.45 42.15 7.99 7.96
2.54 46.39 8.10
2.94 40.19 7.72
5.05 2.95 37.91 41.00 7.76 7.77
?.87 44.89 7.82

-------
                                                  TABLE 13 (continued)
                                 %  Volatile  Sol ids
VO
Date and
Identification



Nov. 13, 1970
VI-B-H-25#










Nov. 25, 1970
VI-A-H-IOO*










Dec. 9, 1970
VI-B-H-IOO*




Each
Disc Sample
IV 76.30
76.44
76.62
1 88.67
89.36
89.36
II 87.60
87.93
87.32
Ml 86.70
86.18
86.50
IV 85.30
84.56
84.79
1 89.74
88.78
89.19
M 87.64
88.18
88.31
III 86.42
87.50
86.48
IV 84.29
85.24
83.93
1 87.50
89.37
90. 1 4
II nr* « *~
1 88.96
87.66
85.80
Avg.

76.45

89.12


87.62


86.46


84.88

89.24


88.04


86.80


84.49

89.00


87.47

Each
Samp 1 e Avq .
4.34
3.67 3.97
3.89
1.77
1.87 1.82
1.81
2.22
2.04 2.12
2.09
2.28
2.19 2.24
2.26
2.39
2.46 2.46
2.53
2.02
1.96 2.00
2.01
2.07
2.02 2.12
2.28
2.39
2.40 2.39
2.37
2.79
2.88 2.82
2.80
1.73
1.68 1.76
1.86
2.43
2.37 2.35
2,26
/v x^*.
Each
Samp le
35.18
37.50
37.76
44.89
40.61
44.59
42.90
68.96
50.63
45.71
46.39
48.15
39.52
43. 12
41.14
52.08
45.86
47.18
47.36
49.91
45.45
50.48
43.73
46.61
51.23
48.49
53. 18
42.74
46.16
37.58
43.54
42.00
46.05
Avg.
CJ
36.81

43.36


54.16


46.75


41.26

48.37


47.57


46.94


50.97

42.18


43.86

f Mil
Each
Samp le
7. 13
7.24
7.26
8.92
9.32
8.88
9.39
9.13
9.66
6. 13
9.42
9.50
8.44
8.89
8.78
9.70
9.22
9.22
8.68
9.02
9 05
•S • V*'
9.39
9.43
9.36
8.22
7.87
8.40
9.05
9.12
8.47
8.78
9.14
8.44
i 
-------
      Date and
      Identi fication
      Dec. 22, 1970
       VIl-A-S-25#
00
                        IV
      Jan. II, 1971
       VIl-B-S-25#
                        I I
                        I I I
                        IV

% Volatile
Each
Sample
86. 16
87.78
87.22
84.58
84.97
85.55
88.77
89.47
89.61
87.72
87.89
87.89
87.96
88.86
87.87
87.31
86.97
87.42
86.92
87.88
87.56
90.88
90.63
90.64
92.23
92.48
92.15
89.96
89.92
89.02

Solids

Avg.

87.05


85.03


89.28


87.83


88.23


87.23


87.45


90.72


92.29


89.60

TABLE 13 (continued
% Phosphorus*
Each
Sample Avg.
2.37
2.35 2.29
2.16
2.80
2.81 2.82
2.84
2.09
1 . 86 1 . 93
1.85
2.50
2.35 2.39
2.32
2.20
2.14 2.21
2.29
2. II
2.10 2.09
2.06
2.01
1.98 2.01
2.05
1.58
1.61 l-6i
1.65
1.57
1.65 1.61
1.61
2.09
2.11 2.13
2.20
)
%
Each
Sample
34.54
36.94
45.68
46.01
54.49
44.02
45.53
47.40
48.75
45.77
46.88
45.00
44.66
44.85
44.92
44.93
45.69
47.32
48.41
49.05
47. f4
47.10
46.58
47.44
47.25
48.08
50.85
45.83
47.96
48.82

Carbon

Avg.

39.05


48. 17


47.23


45.88


44.81


45.95


48.20


47.04


48.73


47.55

% N
Each
Samp le
9.45
9.29
9.4!
9.01
9.33
0.38
9.27
8.84
7.90
7.56
7.78
7.46
7.73
7.93
6.67
7.82
7.56
7.22
8.99
8.27
8.61
8.10
7.94
8.09
8.42
8.03
c
6.83
8.95
8.87
9.09
i trogen

Avg.

9.38


9.57


8.67


7.60


7.44

7.53



8.62


8.04

8.23




8.97


-------
\o
vO
Date and
Identification . Disc
Jan. 29, 1971 1
VH-A-S-IOO#
1 1

III

IV

Feb. 12, 1971 1
VII-A-S-IO#

II


1 II


IV


Feb. 24, 1971 1
VIII-A-H-IO#

II


1 II


IV



% Volati
Each
Sample
85.20
85.91

88.77

86.05

87.78
87.23
89.49
89.47
88.65
88.08
86.34
87.39
86.75
89.22
89.67
90.09
88.87
90.02
89.46
89.31
88.17
89.60
88.39
86.94
87.76
87.54
87.88
86.31 .

TABLE 13 (continued)
le Solids % Phosphorus* % Carbon
Each
Avg. Sample
85.20 .81
.81
85.91 .73
.73
88.77 .67
.64
86.05 .84
.96
1.87
88.16 2.15
2.18

88.73


86.82


89.66


89.45


89.03

.67
.73
2.06
.,96
.90
.89
.63
.91
.87
.67
.68
.81
.84
.87
.85
2.16
87.70 2.24
2.09
2.16
87.24 2.16



Avg.
1.81
1.73

1.66

1.90


2.07


1.82


1.92


1.80


1.72


1.85


2. 16

2.16



Each
Samp le
47.74
48.30

47.89
46.69
48.41

49.97
49.56
49.24
48.41
48.82
47.99
50.86
47.06
50.64
49.92
47.89
51.00
49.81
50.59
48.54
50.40
48.79
49.28
45.71
49.48
45.38
44.31
53.96
46.41
55.05

Avg.
47.74
48.30

47.29

48.41


49.59


48.41


49.52


49.60


49.65


49.49


46.86


49.93


% Nitrogen
Each
Samp 1 e
9.30
9.31

9.35

9.39

9 37
* • -x i
9.52
9.48
8.89
9.26
9.57
9.41
9.39
9.56
9.46
9.94

6.86
8.67
7.91
9.04
9.85
10.34
8.50
10.74

10.00
9.09



Avg.
9.30
9.31

9.35

9.39


9.46


9.24


9.45

9.70



7.81


9.74

9.62


9.54




-------
                                                  TABLE 13 (continued)
     Date and
     Identification

     March 10, 1971
      VI II-B-H-IO*
o
o
     March  26,  1971
       IXL-A-H-IO#
      April 14, 1971
       IXL-B-H-IO#
% Volati 1
Each
Disc Sample
1 87.69
87.78
87.95
II 85.67
85.66
85 31
\J «^ • — ' l
III 85.65
85.15
84.36
IV 88.28
87.98
87.22
1 89.68
92.02
91 .88
I 1 89.82
91.43
91 .24
III 88.27
89.60
89.84
IV 90.17
88.93
90.03
I 89.64
90.05
90.04
M 89.80
89.78

90.27
III 90.14
90.73
90.65
e Sol ids
Avg.
87.81
85.55

85.05

87.83

91. 19

90.83
89.24

89.71

89.91
89.95

90.51

% Phosphorus % Carbon
Each
Samp le
1.43
1 .44
1.39
2.08
2.45
2.14
2.13
1.82
2.14
1.90
2.01
1.99
.24
.48
.78
.49
.63
.45
.80
.74
.84
.84
.65
2.04
.22
.27
.23
.26

.38
.45

Each
Avq. Sample
43.73
1.42 45.08
44.51
44.68
2.22 46.89
45.49
45.28
2.03 46.11

48. 19
1.97 47.63
46. 19
52.03
1.50 49.93
51.28
46.52
1.52 50.96
47.70
44.21
1.79 46.28
48.41
47.44
1.84 46.98
46.76
1.245 46.18
1.245 45.67

1.415 45.52

Avg.
44.44
45.69

45.70

47.34

51 .08

48.39
46.30

47.06

46. 18
45.67

45.52

TO IN i T r
Each
Samp 1 e
8.63
8.80
8"Z C.
.-56
8.65
8.82
8.91
8.42
8.56
81 7
. 1 /
8 A ~I
.47
8.38
81 /-\
. 1 0
10. 17
9.82
I/"\ /*\ 1
0.01
10.25
9.98
9.78
91 f\
. 1 0
10.40
9.48
8.06
9.95
9j- ^
.63
10. 16
10.28
10. 13
9.93
9.84
9Q~7
* O /
9.53
9.79
9.71

ogen
Avg.
8.60
8.79

8.38

8.32

10.00

10.00
9.66

9.21

10.19
9.88

9.68


-------
TABLE 13 (continued)
% Volatile Solids • % Phosphorus % Carbon % Nitrogen
Date and
Identification Disc
IV


April 30, 1971 I
IX.-A-H-50#
L
1 1


III


IV


May 19, 1971 1
IX,-A-H-IOO#
L
1 1


III


IV


June 4, 1971 1
X.-A-H-IO#
L
Each Each
Sample Avg. Sample
88.07
88.63 88.54
88.93
90.88
90.60 90.34
89.55
89.25
88.34 88.63
88.29
88.44
88.15 88.05
87.56
89.71
88.39 88.92
88.67
92.30
92.11 92.05
91.74
91.75
91.21 91.34
91.05
90.31
90.03 90.22
89.43
91.20
89.87 90.58
90.66
90.73
90.20 90.59
90.83
.33
.40

.17
.16
.15
.40
.18
.25
.37
.46
.35
.55
.49

.07
.00
.12
.35
.32
.28
.50
.52
.06
.62
.56
.58
.57
.51
.65
Each
Avg. Sample
1.365 46.42


51.23
1.16 51.38
52.24
56.04
1.28 52.39
52.66
48.94
1.39 50.92

1.52 52.05
50.83
50.74
49.43
1.06 52.65
51.94
49.60
1.32 47.93
50.55
51.38
1.36 50.95
52.20
46.87
1.59 45.68
50.48
46.01
1.58 44.98
44.86
Each
Avg. Sample
46.42 9.92
9.78
9.93
10.21
51.62 9.99
10.11
9.83
53.70 9.70
10.32
49.93 9.54
9.53

9.04
51.21 8.83
9.21
9.03
51.34 9.36
9.01
9.60
49.36 9.70
9.40
9.06
51.51 10.35
9.71
9.60
47.68 9.40
9.43
9.51
45.28' 9.28
9.32

Avg.

9.88


10.10


9.95

9.54



9.03


9. 13


9.57


9.71


9.48


9.34


-------
                                                    TABLE 13 (continued)
o
N)
% Vo 1 at II
Date and Each
Identification Disc Sample
1 1 89.78
89.4!
89.52
1 1 1 88.61
88.51
88.52
IV 85.76
85.92
85.38
June 23, 1971 1 89.99
X,-A-H-IOO# 90.08
L 90.08
II 89.74
90.28
89.64
III 87.42
111 w » • ^ *-
86.05
86. 16
IV 83.87
83.57
84.04
July 2, 1971 1 83.45
VIII . -A-H-IO# 89.96
L 90.41
II 88.66
88.56
88.25
III 95 74
III & -* * * ~
89.05
88.52
IV 88.28
88.10
88.12
e So lids % Phosphorus % Carbon % Nitrogen
Each
Avq. Sample
1
89.57 1
1
2
88.55 2
2
.73
.86
.74
.19
.06
.28
2.72
95.69 2
2
90.05

89.89

.60
.81
.35
.56
.30
.53
.64
.61
2.45
86.54 2.39
2.48
2.94
83.83 2.91
3.15
87.94

88.44


91.10


88. 17

.52
.55
.48
.59
.72
.75
.77
.80

.95
.90
.98
Each
Avg. Sample
44.89
1.78 44.85
43.84
44.93
2.18 45.94
41.32
41.98
2.71 46.65
43.67
45.00
1.40 47.32
48.27
45.26
1.59 45.00
45.52
43.88
2.44 45.04
43.69
43.35
3.00 43.88
43. 13
43.99
1.52 47.40
53.40
45.00
1.69 47.14
45.15
1.79 43.65
43.61
45.45
44.85
1 . 94 41.81
44.2!
bach
Avq. Sample
9.96
44.86 9.52
9.67
10.05
44.06 9.95
9.84
9.23
44.10 9.25
9.47
8.31
46.86 8.37
8.98
8.41
45.26 8.37

8.67
44.20 8.53
7.95
8.58
43.45 8.62
8.24
10.89
48.26 10.42
10.52
9.91
45.76 10.53
9.62
10.02
44.24 11.48
10. 10
9.68
43.62 9.72
9.41
Avg.

9.72


9.95


9.32

8.55
8.39



8.38


8.48

10.61

10.02


10.53


9.60


-------
                                                  TABLE  13  (continued)

                               %  Volatile Sol ids     % Phosphorus
Date and
Identification Disc
July 19, 1971 1
VIII.-A-H-IOO#
L.
ii


in


IV


Each
Samp 1 e
87.24
87.13
87.27
82.37
82.92
83.12
81.06
80.82
80.71
80.52
80.00
79.82

Avg.

87.21


82.80


80.86


80.1 1

Each
Sample
1.73
1.66
1.51
2.00
2.17
2.28
2.67
2.81
2.69
2.91
2.21


Avg.

1.63


2.15


2.72

2.56


% Carbon
Each
Samp 1 e
44.96
44. 18
44.78
42.49
43.94
43.35
45.94
40.99
44.04
39.68
41.55
40.99

Avg.

44.64


43.26


43.66


40.74

                                                                                         % Nitrogen
                                                                                      Each
                                                                                      Sample
                                                                                            8.99
                                                                                            8.03

                                                                                            8.55
                                                                                            8,37

                                                                                            7.66
                                                                                            8.15
                                                                                            7.99
                                                                                            7.73
                                                                                            7.36
                                                                                            7.39
                                                                                                 8.46
                                                                                                 7.93
                                                                                                 7.49
o
UJ
      **
* On dry weight basis
+ Composite from four discs
  All these slime samples were taken from the settling cones
  Test runs included in statistical  analysis
  After washing with deionized water
  Before washing with deionized water
  Problem with laboratory analysis

-------
                               TABLE 14
                    PERCENT CALCIUM AND MAGNESIUM
                            IN DISC SLIMES
Date and
Identification

October 9, 1970
V-B-S-IOO
November  13,  1970
YI-B-H-25
 November 25,  1970
 VI-A-H-IOO
 December 9,  1970
 VI-B-H-IOO
                                          Percent on Dry Weight Basis


Disc
1


II


III


IV


1

II


III


IV


1

II


III


IV


I


II

Ca
Each
Sample
0.51
0.50
0.42
0.48
0.50
0.42
0.53
0.49
0.58
0.39
0.46
0,37
2,18
1.58
1,86
1.76
1,82
2.17
2.33
2.20
3.13
2.71
2.83
2.12
2.24
2.84
2.61
2.44
3.55
3.46
1,92
4.14
3.67
4.22
1.49
1.42
1.48
3.25
3.07
Mq
Each
Ave . Samp 1 e Ave.

0.48


0.47


0.53


0.41

1.88


1.81


2.23


2.89

2.18 .21 .215
.22
.25
2.63 .21 .22
.20
.23
2.98 .26 .25
.26
.26
4.01 .23 .25
.26

1.46


2.98
                                           2.61
                              104

-------
                          TABLE 14 (continued)
                                          Percent on Dry Weight Basis
Date and
Identification
December 22, 1970
VII-A-S-25
January II, 1971
VM-B-S-25
January 29,
VII-A-S-IOO
1971
February 12, 1971
VII-A-S-IO


Disc
III


IV


1


II

III

IV

1


II


III


IV


1

II

III
IV

1


II


Ca
Each
Samp 1 e
3.73
3.39
3.30
4.39
4.27
4.70
.71
.66
.69
.97
.79
.71
.44
.92
.83
.71
.75
.70
1.29
1.12
1.24
1.99
2.00
2.15
2.89
2.88
2.83
.40
.41
.81
.83
1.04
1.66
1.53
.36
.48
.38
.80
.84
.79


Ave.

3.47


4.45


.69

1.88

1.58

1.88


.72


1.22


2.04


2.87

.41

.82

1.04
1.60


.41


.81

Mg
Each
Samp 1 e






.23
.20
.19
.22
.22
.19
.17
.15
.19
.17
.18
.17
.13
.13
.13
.12
.11
.15
.12
.14
.13
.105
.106
.08
.09
.08
.08
.08
.11
.13
.12
.09
.10
.10


Ave.







.21

.22

.18

.17


.173


.13


.13


.13

.106

.09

.08 •
.08


.12


.10

                            105

-------
   TABLE 14 (continued)
              Percent on Dry Weight Basis

Date and
Identification Disc
III


IV


February 24, 1971 1
VIM-A-H-IO

II


III


IV


March 10, 1971 1
VMI-B-H-IO

II


III


IV


March 26, 1971 1
IX.-A-H-IO II
L

III


IV


Ca
Each
Samp 1 e
.60
.12
.10
.15
.78
.41
.79
.90
.76
.74
.82
.78
2.17
2.25
2.12
1.90
1.99
2.24
3.50
3.99
3.71
3.03
2.95
2.95
4.28
3.94
3.85
2.46
2.54
3.30
.57
.70
.69
.71
.64
.70
.75
1.15
1.37
1.56


Ave.

1.11


1.45


1.82


1.78


2.18


2.04


3.73


2.98


4.02


2.77

.57

.70


.70


1.36

Mg
Each
Samp 1 e Ave .
.06
.10
.10
.086
.108 .094
.087
.41 .40
.39

.33 .33


.24
.24 .243
.25
.26
.26 .27
.28
.31
.32 .31
.30
,40 .39
.38

.42
.36 .39
.38
.34
.36 .35

.01 .01






.35
.35 .34
.31
106

-------
TABLE 14 (continued)
                 Percent on Dry Weight Basis
                     Ca
Mg
Date and
Identification
April 14, 1971
IX.-B-H-IO
I
L.









April 30, 1971
IX.-A-H-50
L








May 19, 1971
IX.-A-H-IOO
L








June 4, 1971
X.-A-H-IO
L





Each
Disc Sample
1 .75
.82

.89
II .84
.89
.91
III .19
.17
.11
IV .19
.06
.17
1 .79
.82
.85
II 1 .02
.86
III .11
.06
.06
IV .36
.37
.43
1 .84
.82
.80
II ,04
.07
.07
Ml .17
.20
IV .47
.50
.44
1 1.59
2,17
2.09
II .58
.70
III .65
.97
.95

Ave.

.82



.88


1.16


1.14


.82

.94


1.08


1.39


.82


1.06

1.18

;
1.47


2.13

1.64


1.86

Each
Sample Ave.
























.30 .29
.27

.37
.30 .33
.32
.34 .34

.31 .32
.32

.23
.22 .22
.21
.23 .23
.23
.25 .24
.24

    107

-------
                          TABLE 14 (continued)
                                          Percent on Dry Weight Basis
                                          	Ca	       Mg
Date and
Identification
June 23, 1971
XL-A-H-IOO
July 2,  1971
VIIIL-A-H-I,0
July  19,  1971
VMIL-A-H-IOO
                            IV
                            IV
Each
Sample
2.78
2.86
2.93
1.75
1.59
1.57
2.30
2.46
2.26
2.84
3.48
3.31
4.05
4.00
4.14
1.46
1.26
1.37
2.12
1.96
2.00
1.89
2.02
1.92
2.94
2.71
3.12
3.00
4.04
4.37
4.26
4.50
4.77
4.54
6.51
5.36
5.04

Ave.

2.86


1.64


2.34


3.21


4.06


1.36


2.03


1.94

2.88

3.06


4.22


4.60


5.20

Each
Sample
.25
.28

.24
.20
.24
.23
.19
.22
.26
.26
.27
.26
.31
.30
.24
.25
.24
.30
.34
.30
.29
.30
.28
.29
.29
.29
.31
.37
.39
.31
.37
.47
.38
.46
.39
.34

Ave.
.27



.23


.21


.26


.29


.24


.31


.29

.29

.30


.36


.41


.40

                             108

-------
                                             TABLE  15
                                    ANALYSIS OF CHANNEL  SLIMES
                                         PRELIMINARY RUNS
Date and
Jjdent ? f i cat f on

March 26, 1971
 Mllk(24 g./SOL)
 only feed

April 16, 1971
 Ml Ik only,
 4.0 ml/mln.
 Mid-slope
% Volatile Sol ids     % Phosphorus
Carbon
% Nitrogen
June I, 1971
 Mllk(36g/80L),
 80ml N.B. solfn
 <450mg/ml), 20ml
 Na2HPO (20mgP/ml
Channel Each Sample Avg. Each Sample Avg. Each Sample Avq. Each Sample Avq.
Comp . * 91.4
91.2 91.3
91.3
I

II

1 1 1


IV


.24
.25
.40
.21
. 14
.20
. 14
.22
.20
.09
. 17
. 19
. 19
1 2. 15
1.89
M 2.00
2.02
1.96
111 88.6 88.6 1.97
2.10
2.04
IV 87.8 87.3 2.21
86.8 2.09
2.26
49.50
1.30 49.44 49.53
49.66
1. 175

1. 17


1. 16


1. 18

2.02


1.99


2.04


2. 19

10.48
9.90
10.26







10.73
10.74



8.65


8.94
9.02

8.07
8. 18


10.21








10.74




8.65


8.99


8. 13


  Average percent of calcium in slime = 0.62$.

-------
                                             TABLE 16
                                    FEED AND EFFLUENT  ANALYSES
                       REMOVAL-STORAGE COMPARISON RUNS ON CHANNEL APPARATUS
                                                          Feed (ma/I)
Run       Comments

I     Low slope.  4 ml/min. 24 mq milk
     so I ids/SQL.  50 ml stock P~sol'n.
     (12 mg P/ml=7.5 mg P/L + milk)
                                                           Effluent (mg/I)
                                            Date    Phosphorus  COD
                                          4-19-71
                                          4-20-71
                                          4-22-71
II    Mid slope.   Same feed as
     Same rate as 1.
I.
4-27-71
                                          4-28-71
                                          4-29-71
                                          5-4-71
                         9.05
                         8.95
                         8.49
8.64
                         8.40
                         8.17
                         7.41
                      318    15.2
                      265    15.7
                      314    15.4
                                                                301
                      339
                      313
                      331
16. I
                16.4
                13.8
                14.5
                          I
                         I 1
                        I I I
                         IV
                          I
                         II
                        I II
                         IV
                          I
                         11
                        III
                         IV
                                                                               11
                                                                                IV
                                                                                iv*
                                                                                IV
Phosphorus
9.71
9.85
8.98
9.46
9.44
9.44
8.83
9.63
8.49
8.58
14. 10
8.38
8.56
9.05
7.72
8.47
8.38
8.59
8.09
7.96
8.13
8.15
7.89
7.80
8.80
8.36
8.31
9.08
COD
343
326
289
275
219
199
170
141
285
281
273
240
255
238
188
265
291
283
266
261
230
224
178
167
254
239
181
164
Nitrogen
22.3
21.9
19.3
20.3
16. 1
13.9
12.1
11.9
17.4
17.7
19.0
17.9
14.5
14.7
1 1.7
1 1.4
14.7
13.8
14.2
13.7
10.0
9.2
9.8
8.9
14. 1
13.3
1 1.9
1 1.3

-------
                                             TABLE 16 (continued)
Run
Comments
                                                           Feed  (mg/l)
                                                                                Effluent (mg/l)
 II  Mid slope.  4 ml/min.  36 g
     glucose, 80 ml Nutrient broth
     sol'n. (450 mg/ml), 20 ml
             (20mgP/ml).
IV   Low slope.
     as 111.
       Same feed and rate
Date
5-5-7 1



5-6-71



6-8-71



6-9-7 1



6-11-71



6-14-71



6-15-71



6-18-71



Phosphorus "COD Nitrogen Channel
8.83 365 15.4 1
1 1
1 1 1
IV
8.83 326 14.7 1
1 1
1 1 1
IV
10.03 780 55.0 1
1 1
1 1 1
IV
10.00 932 59.5 1
1 1
II 1
IV
9.38 924 61.8 1
1 1
1 1 1
IV
7.20 959 60.2 1
1 1
1 1 1
IV
10.19 928 64.4 1
1 1
1 1 1
IV
10.91 935 63.4 1
1 1
1 1 1
IV
Phosphorus
9.05
8.85
8.95
8.93
8.45
8.38
8.59
9.10
9.07
9.07
8.78
9.94
7.94
9.13
8.43
8.58
9.05
9.41
8.31
10.03
9.78
6.07
8.72
6.53
1 1.30
9.07
10.25
10.06
26.08
10.14
9.32
10.06
COD
298
279
270
262
249
227
193
187
804
780
760
812
772
772
788
796
824
768
820
,779
734
827
850
858
928
802
858
771
862
838
834
798
Nitrogen
15.9
15.5
15.7
16.3
10.4
10.2
1 1 . 1
1 1.4
55.7
60.7
56.0
58.5
54.0
55.5
53.9
56.7
57.8
59.3
55.7
56.7
52.3
60.4
59.3
52.3
64.6
57.2
56.5
56.2
59.9
56.9
55.7
52.0

-------
                                                  TABLE 16 (continued)
      Run
     Comments

High slope.  Same feed and rate
as III.
                                                                Feed (mg/l)
                                                                                     Effluent  (mg/l)
N>
      VI
Mid slope.
milk, 15 ml
80ml NH4CI(48mgN/
ml/min.   Separate tubing
col lection.
75g glucose, 25 g
Na,HPOd(20mgP/ml),
8mgN/mT)/80L.  5.4
             for
      VII   High slope.
           rate as VI.
             Same feed and
Date
6-21-71



6-22-71



6-23-71



7-6-71



7-7-71



7-8-71



7-14-71



7-15-71



Phosphorus COD Nitrogen Channel
9.94 942 60.7 1
1 1
II 1
IV
10.38 880 60.2 1
II
1 II
IV
13.54 935 1
1 1
1 1 1
IV
6.98 1356 70.9 1
1 1
1 II
IV
7.85 1341 65.6 1
1 1
1 1 1
IV
5.42 1342 70.0 1
II
1 1 1
IV
5.62 1324 62.7 1
1
1 1
V
5.46 1336 58.8 1
1
1 1
V
Phosphorus
8.34
10.58
10.38
9.35
10.68
9.63
10.03
8.52
8.52
9.69
13. 13
9.08
5.76
6.29
5.94
5.32
6.43
5.28
5.79
5.22
5.30
5.00
3.62
3.02
5. 10
4.30
4.46
4.06
5.31
4.50
4.75
3.44
COD
770
742
774
770
701
748
788
741
719
781
765
715
1290
1310
1325
1193
1255
1 189
1015
1255
1272
1280
1 1 17
974
1284
1260
1244
1208
1304
1280
1216
1128
Nitroqen
50.4
55.4
56.5
57.4
46.3
52.3
56.0
53.7
*
*
*
*
59.3
66.6
63.7
63.5
61.0
57.9
46.7
44.9
66.9
57.8
54.8
45.9
65.5
62.7
61.04
64.6
64. 1
56.8
56.5
50.4

-------
                                             TABLE  16 (continued)
Run
          Comments
                                          7-16-71
VI II Low slope.
     as VI.
                 Same feed and rate
7-20-71
                                          7-21-71
                                          7-22-7'I
                                                           Feed  (mg/l)
                                            Date    Phosphorus   COD  Nitrogen  Channel
                                                                                          Effluent (mg/l)
                                                       5.73
                     1383    57.4
5.47    1298    63.2
                                                       5.73     1317    66.3
                                                       5.94    1309    70.0
                           I
                          I I
                          I I
                          IV
Phosphorus
5.03
4.58
4.21
3.98
5.30
5.01
4.79
4.53
4.51
4.51
4. 15
4.06
4.51
4.41
4. 14
3.61
COD
1312
1288
1272
1 161
1275
1294
1279
1228
1262
1215
1219
1 183
1238
1 183
1 159
1088
Nitrogen
65.5
62.4
59.3
58.8
56.5
59.9
56.5
61.0
59.9
59.3
57.6
62. 1
68.6
60.2
59.9
48.7
*Ana-|ysis problem

-------
            TABLE 17
   ANALYSIS OF CHANNEL SLIMES
REMOVAL-STORAGE COMPARISON RUNS
Volatile Sol ids.     % Phosphorus
                                         % Carbon
% Nitrogen
Run Date Channel Each Sample Avg. Each Sample
t 4-23-71 1

II
III

IV
II 5-20-71 1

1 1


III 87.0 87.0


IV 85.4 85.4


III 6-1 1-71 1


II 93. 1 93. 1


1 1 1


IV


.35
.34
.39
.47
.54
.43
.25
.21
.24
.13
.22
.16
.23
.27
.14
.25
.02
.72
c *y
.53
.51
.57
.52
.55
.59
.61
.50
.56
.77
Avg. Each Sample
1.34

1.39
1.50

1.43
1.23


1.20


1.22


1.14

If *"i
.62


1.53


1.58


1.61

Avg. Each Sample








1 1.21


10.36
10.51
10.88
10.40
10.58
10. 20



8.97
9.58

10.06
9.74




Avg.








1 1.21



10.58


10.40




9.28


9.90






-------
Ul
                                                  TABLE  17  (continued)



                                   % Volatile Solids     % Phosphorus
% Carbon
%  Nitrogen
Run Date Channel Each Sample Avg. Each Sample Avg. Each Sample Avg. Each Sample Avg.
IV 6-18-71 1

II


IN


IV


V 6-25-71 1


II 88.72 88.57 ;
88.41
*•
III 91.65
88.92 89.73
88.64
IV 90.00
91.56 90.61
90.26
VI 7-9-71 1


.32
.62
.77
.60
.74
.88
.63
.57
.62
.76
.85
.58
.81
.83
?.20
.86
1.20
.68
.63
.85
.68
.85
.53
.97
.98
.82
II 86.13 86.13 2.57
2.27
1.85
III 88.47 88.47 1.76
1.72
1.71
1.47


1.70


1.69


1.74


1.74


2.09


1.73


1.69


1.92


2.23


1.73



9.77


8.80


9.71
9.99

9.63


10.04
10.01
9.64
10.34
10.40
10.30
1 1,53
11.38




9.36
8.26
8.14
10.39
10.22



9.77


9.90


9.85


9.63



9.90


10.35

1 1.46






8.59

10.30



-------
Run      Date
VII    7-16-71
VIM   7-23-71
*Analysis problem.
                                             TABLE  17 (continued)
                              % Volati le Sol ids     % Phosphorus          % Carbon	     % Nitrogen
                     IV
                     I II
                     IV
                     I I
                      II
Each Sample
84.66








89.18
90.77







88.35
89.24

87.13
84.51
85.89
Avg. Each Sample
84.66 1.69
1 .81
1.62
*
2.30
1.92
1.82
2.27
2.23
89.98 *;°<
2.02
1.66
1.98
2.05
1.95
2.01
2.00
88.80 1.79
1.71
1.71
2.01
85.84 2.01
2.13
Avg. Each Sample

1.71


2.11


2.11

2.10


1.90


1.99


1.74


2.05

Avg. Each Sample
10.62
1 1.46







9.90
9.60
9.78
11.12


9.28


9.79
9. 13
10.25
9.93
9.39
9.78
Avg.
1 1.04








9.76

11.12


9.28



9.72


9.70


-------
                                             TABLE 18
                                    FEED AND EFFLUENT ANALYSES
                             ULTRAVIOLET STUDIES ON CHANNEL APPARATUS
Run   Date
I    8-11-71 Mid
     Comments
Mi Ik, Glucose, Na HPO
Cl for feed as last
study (removal-storage
comp.)
    8-12-71
II  8-16-71 Mid   Same feed as I.
Feed (mg/l) Effluent (mg/l)
Time Phosphorus COD Nitrogen Channel Phosphorus COD N
Initial 5.61 !2I5 60.7

1

UV-I 5.61 1214 58.5

|

UV-2 5.44 1206 59.0

1

UV-I 9 1/2 5.46 1 198 52.9

1

UV-26 5.44 1182 64.4

1







\



\



3.22 897
2.74 782
2.18 741
/ 1.46 602
5.28 1072
6.12 1052
6.43 1092
/ 6.76 1028
5.32
5.44
5.68
/ 5. 18
5.31
5. 17
5.30
V 5. 10
1 4.99
1 4.72
1 5.44
V 4.51
133
060
084
028
194
133
190
092
161
121
186
145
Initial 5.61 1135 77.5 1 3.61 1131
1 1.63 1015
II 0.96 840
V 1.26 832
UV-I 5.24 1215 57.6 1 6.00 1235
1 6.79 1219
1 1 7.33 1171
V 7.91 1199
i trogen
*
40.8
39.4
*
50.6
52.6
53.2
59.0
58.2
52.0
51.8
55.7
59.6
56.8
61.3
55. 1
56.5
53.7
58.8
53.7
54.6
53.7
44.2
42.0
78.9
66.0
64.4
71.4

-------
                                                   TABLE 18 (continued)
      Run    Date  Slope      Comments




          8-17-71
                                                                Feed (mg/l)
Effluent (mg/l)
o>
          8-18-71
          8-1 9-71
Time Phosphorus COD Nitrogen Channel Phosphorus COD
UV-19 4.20 1183 46.4

1

UV-24 5.44 1201 56.0

1

UV-48 5.04 1203 63.2

1

UV-72 5.15 1262 55.4

1



\



\



\



4.27 996
4.17 999
3.76 980
/ 3.62 972
5.44
5.28
4.94
/ 4.94
5.31
5.18
4.99
/ 5. 13
5.65
6.25
5.94
V 6.02
217
185
161
161
170
170
154
I5C
25
-------
                                                   TABLE 19
                                          FEED AND EFFLUENT ANALYSES
                                    STARVE-KILL STUDY ON CHANNEL APPARATUS
\o
      Run  Date   Slope
     Comments
                                                                Feed (mg/l)
                                                                        Effluent  (mg/l)
                        study.  Same rate as
                        ultraviolet study
          8-24-71       24 hrs. with H20 feed
          8-25-71
24 hrs. with hU) feed.
Put on UV light, stiI  I
with HO feed.
      II  8-30-71 Mid   Same feed and rate.
          8-31-71       24 hrs. with H20 feed.
          9-1-71
24 hrs. with H?0 feed.
Put on UV light, stiI  I
with H20 feed.
      Ill 9-8-71  Mid   Same feed and rate
Time Phosphorus COD Nitrogen Channel Phosphorus COD
Initial 2.0 1331 58.0
1
II
1
5.61 1296 *
1
1 1
1
UV-2 5.42 1318 61.0
1
1 1
1
Initial 5.61 1303 59.3

1

5.22 1128 53.7

1

UV-2 5.61 1259 59.3

1

Initial 5.31 1325 61.0
1
1 1
1
1 0.374 1173
1 0.289 1022
1 	 812
V 	 761
1 4.51 1158
1 4.87 923
1 2.95 796
V 2.56 642
1 6.05 1207
1 6.52 1065
1 7.07 961
V 7.32 823
4.39 1136
3.68 996
3.10 784
V 2.41 791
3.52 902
3.39 793
2.23 524
V 1.93 470
6.39 1148
7.30 1086
7.82 954
V 8.78 926
1 3.71 1180
1 2.33 1060
1 0.91 935
V 0.30 799
Nitrogen
62
52
46
47
57
49
42
45
58
56
54
53
55
54
45
48
45
41
34
32
57
56
56
57
57
51
42
46
.4
.3
.4
.0
.6
.5
.0
.3
.2
.2
.0
.7
.7
.0
.9
.7
.6
.7
.1
.4
.4
.8
.0
.6
.6
.5
.2
.4

-------
                                             TABLE  19 (continued)
Run  Date   Slope
      Comments
                                                          Feed (mg/l)
 Time  Phosphorus  COD
                                                                         Effluent (mg/l)
    9-9-71
    9-10-71
    9-11-71
 24 hrs.  with H20 feed.
 24 hrs.  with H~0 feed.
 Put on UV Iighf, stiI I
 with H20 feed.

 24 hrs.  with H?0 feed
 and UV Iight.
UV-2
UV-24
IV  9-14-71 Low   Same feed and rate.
    9-15-71
    9-16-71
    9-17-71
 24 hrs.  with H20 feed.
 24 hrs.  with  HO feed.
 Put on  UV IighT, stiI I
 with H20 feed.

.24 hrs.  with  HO feed
 and UV  light. Z
UV-2
UV-24
          5.56
5.76
5.46
                            Initial    1.95
          5.62
5.73
5.74
        1298    60.7
1282    59.9
1252    57.1
                  1290
        1255
1288
1283
                67.7
        60.7
60.2
60.4
anne
L
1
1
V
1
1
1
V
1
1
1
V










V



V



V
1 Phosphorus
4.82
4.29
3.36
2.38
5.31
5.78
6.22
6. 10
5.08
5. 17
5. 14
4.74







3.59
2.60
1.75
	
4.62
3.84
3.34
3.81
6.07
4.62
4.88
4.62
COD
1 120
991
887
741
1236
1083
1007
892
1 154
1101
1078
961
1237
1 1 Af,
I 1 HO
QKfl
17 J\J
--CA
1 J"
955
849
699
481
1 190
1 167
1088
888
1283
1237
1 153
1163
Nitrogen
54.3
49.8
44.2
45.0
53.7
52.8
53.4
47.6
54.3
50.4
48.7
44.8

fi9 A
Of. . 1
R~7 A
j / . *t
51 S
1 . J
47.3
39.2
32.7
27.4
56.2
54.3
55.1
61.6
58.8
53.7
54.3
50.6

-------
                                             TABLE  19  (continued)
     Run   Date   Slope
                   Comments
                                                                Feed  (mg/l)
     V   9-20-71  Low   Same  feed  and  rate.
9-21-71       24 hrs. with H20 feed.
                            Time  Phosphorus  COD

                           Initial   5.61    1243    62.4
                                                             5.33    1279    59.6
                                                                                     Effluent (mg/t)
Is)
         9-22-71
         9-23-71
24 hrs. with H?0 feed.
Put on Light with H20
feed.

24 hrs. with H,0 feed
and UV Iight. *
                                         UV-2
      VI   9-27-71 High  Same  feed and rate.
          9-28-71
          9-29-71
          9-30-71
24 hrs. with H00 feed.
              24 hrs. with HO feed.
              Then put on UV IIght
              with H20 feed.

              24 hrs. with H~0 feed
              and UV light.
                           UV-2
                                     4.01    1303    55.4
                                         UV-24     5.24    1303    58.2
                                         Initial    5.25    1277    59.3
                                                   5.43    1293    59.3
                                     5.13    1238    48.1
                           UV-24     5.31    1255    58.8
                                                                                      IV
                                                                                      IV
Phosphorus
3.94
3.48
3.87
3.61
5.28
4.77
4.37
3.27
1.26
1.22
1.10
1.07
4.33
3.36
3.62
2.74
4.51
4.33
3.82
3.08
4.50
4.47
3.81
3.50
4.98
4.95
5.61
5.59
4.29
4.09
5.90
3.71
COD
1218
1297
1 148
1115
1 177
1076
983
902
1233
1210
1171
1108
1260
1214
1187
1 148
1227
1219
1127
1103
1159
1125
1016
945
163
146
138
171
179
179
330
217
Nitrogen
58.2
59.0
60.2
61.3
56.2
*
49.8
45.3
52.0
50.4
53.7
50.9
54.8
53.7
53.2
51.5
57.6
56.0
54.6
56.5
55.4
52.6
49.8
47.0
53.7
55.1
56.8
57.1
53.7
53.7
60.4
54.8

-------
                                               TABLE 19 (continued)
     Run    Date  Slope
                  Comments
 Time  Phosphorus
                                                                Feed (mg/I)
                                                                                    Effluent (mg/l)
 NJ
 M
      VII   10-4-71  High Same feed and rate.
           10-5-71       24 hrs.  with H20 feed.
10-6-71       24 hrs.  with H,,0 feed.
             Put on UV light, with
             H20 feed stiI I  on.

10-7-71       24 hrs.  with HO feed
             and UV Iight.
                                        Initial    6.78    1285    59.3
                                                  5.46    1248    59.9
                                                   UV-2      5.58    1292    60.2
UV-24     5.61     1276    61.0
Phosphorus
5.15
5.18
3.58
3.49
4.09
3.50
3.15
2.03
5.74
5.94
5.70
6.64
4.96
4.96
4.69
4.79
COD
1165
1140
1012
942
1060
971
838
475
1180
1 164
1101
1093
1 184
1168
1 105
1041
Nitrogen
56.2
53.2
50.6
47.0
52.3
41.4
45.3
39.7
55.7
57.1
55.4
57.6
54.3
52.0
52.0
52.0
I
3
      *Analysis problem.
I
8

-------
   SELECTED WATER
   RESOURCES ABSTRACTS
   INPUT TRANSACTION FORM
                      /. Report No.
                         3. Accession No.
                                           W
  4.  Title
         PHOSPHORUS REMOVAL BY TRICKLING  FILTER SLIMES
  7. Author(s)

  	Zanoni, A. E.
  9. Organization

         Marquette  University
         Department of Civil Engineering
         Milwaukee,  Wisconsin
  12. Sponsoring Organization

  15. Supplementary Notes

         Environmental Protection Agency report number,
         EPA-R2-73-279, July 1973.
                                           5. Report Date
                                           6.
                                           8. Performing Organization
                                             Report No.

                                          10. Project No.

                                              17010 DZG
                                          //. Contract/Grant No.

                                              17010 DZG
                                          13, Type of Report and
                                             Period Covered
  16. Abstract
 A rotating disc apparatus was constructed so that disc  speed could be varied.   Orgaaisms
 that developed on  the disc surface,  in response to various nutrient solutions,  could be
 harvested and analyzed.  Correlations  were.attempted between mineral composition of the
 feed solutions and the phosphorus and  nitrogen content  of the resulting  slimes.

 An inclined channel  apparatus was also constructed and  evaluated to differentiate
 physical or chemical mechanisms from biological mechanisms of phosphorus uptake.  The
 angle of inclination was used to measure the kinetic rates before and after inactivation
 of the biological  slime with ultraviolet light.

 With the disc apparatus, limited success of inducing biological uptake of  phosphorus, in
 excess of 1.5 to  2.5 percent of the  cell mass, was obtained.  Statistical  analysis of the
 data indicated that those values above 2.5 percent usually were encountered when the
 medium contained  calcium salts.  Results from the inclined plane growth  chamber showed
 that the limited  phosphorus uptake  that did occur could be related to metabolic activity
 rather than physical sorption or chemical precipitation.
  17a. Descriptors
 *Phosphorus Removal, *Biological  Treatment, Nutrient  Studies
  17b. Identifiers
 #Attached Growth,  Synthetic Media,  Research Apparatus
  17c. COWRR Field A Group  05D
  18. Availability
19. Security Clast.
   (Report)

20. Security Clas*.
  Abstractor   E.  F. Barth
21. No. of
   Page*

22. Price
Send To:
                                                        WATER RESOURCES SCIENTIFIC INFORMATION CENTER
                                                        U.S. DEPARTMENT OP THE INTERIOR
                                                        WASHINGTON. D.C. 20240
              Ja«trtutyon
                                                EpA>
                                 Cincinnati. Ohio
WRSIC 102 (REV. JUNE 1971)
                                                                *U3. GOVERNMENT PRINTING OFFICE: 1973 546-303/16

-------